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Microcolony and biofilm formation as a survival strategy for bacteria
Bacterial communities such as biofilms are widely recognized as being important for survival and persistence of bacteria in harsh environments. Mechanistic models of biofilm growth indicate that the way in which the surface is seeded can effect the morphology of simulated biofilms. Experimental studies indicate that genes which are important for chemotaxis also influence biofilm formation, perhaps by influencing aggregation
Leah R. Johnson
Initial biofilm formation of acetoclastic methanogenic bacteria
The initial rate of acetoclastic methanogenic biofilm formation was significantly higher than rates that have been reported for mixed anaerobic cultures. Biofilm development rate increased with an increase in biomass concentration and specific activity (mg substrate?mg biomass?day) of the mixed liquor biomass. Biofilm concentration also increased with higher specific activity. Results indicate the involvement of polysacchar?ides in acetoclastic biofilm formation.
R. L. Droste; E. Andras; K. J. Kennedy
Biofilm bacteria: formation and comparative susceptibility to antibiotics
The Calgary Biofilm Device (CBD) was used to form bacterial biofilms of selected veterinary gram-negative and gram-positive pathogenic bacteria from cattle, sheep, pigs, chicken, and turkeys. The minimum inhibitory concentration (MIC) and minimum biofilm eradication concentration (MBEC) of ampicillin, ceftiofur, cloxacillin, oxytetracycline, penicillin G, streptomycin, tetracycline, enrofloxacin, erythromycin, gentamicin, tilmicosin, and trimethoprim-sulfadoxine for gram-positive and -negative bacteria were determined. Bacterial biofilms were readily formed on the CBD under selected conditions. The biofilms consisted of microcolonies encased in extracellular polysaccharide material. Biofilms composed of Arcanobacterium (Actinomyces) pyogenes, Staphylococcus aureus, Staphylococcus hyicus, Streptococcus agalactiae, Corynebacterium renale, or Corynebacterium pseudotuberculosis were not killed by the antibiotics tested but as planktonic bacteria they were sensitive at low concentrations. Biofilm and planktonic Streptococcus dysgalactiae and Streptococcus suis were sensitive to penicillin, ceftiofur, cloxacillin, ampicillin, and oxytetracycline. Planktonic Escherichia coli were sensitive to enrofloxacin, gentamicin, oxytetracycline and trimethoprim/ sulfadoxine. Enrofloxacin and gentamicin were the most effective antibiotics against E. coli growing as a biofilm. Salmonella spp. and Pseudomonas aeruginosa isolates growing as planktonic populations were sensitive to enrofloxacin, gentamicin, ampicillin, oxytetracycline, and trimethoprim/sulfadoxine, but as a biofilm, these bacteria were only sensitive to enrofloxacin. Planktonic and biofilm Pasteurella multocida and Mannheimia haemolytica had similar antibiotic sensitivity profiles and were sensitive to most of the antibiotics tested. The CBD provides a valuable new technology that can be used to select antibiotics that are able to kill bacteria growing as biofilms.
Olson, Merle E.; Ceri, Howard; Morck, Douglas W.; Buret, Andre G.; Read, Ronald R.
Biofilm Formation by Gram-Negative Bacteria on Central Venous Catheter Connectors: Effect of Conditioning Films in a Laboratory Model
Human blood components have been shown to enhance biofilm formation by gram-positive bacteria. We investigated the effect of human blood on biofilm formation on the inner lumen of needleless central venous catheter connectors by several gram-negative bacteria, specifically Enterobacter cloacae, Pseudomonas aerugi- nosa, and Pantoea agglomerans. Results suggest that a conditioning film of blood components promotes biofilm formation by these
R. Murga; J. M. Miller; R. M. Donlan
Enhancement of nitrifying biofilm formation using selected EPS produced by heterotrophic bacteria
The possibility of enhancing nitrifying biofilm formation rate with the aid of selected EPS produced by heterotrophic bacteria was investigated. When EPS production characteristics were examined for four kinds of heterotrophs isolated from a domestic wastewater treatment reactor, two strains obtained from biofilms (B1, B2) exhibited a higher polysaccharide production rate than those from suspended flocs (A1, A2). Among EPS
S. Tsuneda; H. Hayashi; J. Jung; A. Hirata
Adhesion and biofilm formation on polystyrene by drinking water-isolated bacteria
This study was performed in order to characterize the relationship between adhesion and biofilm formation abilities of drinking\\u000a water-isolated bacteria (Acinetobacter calcoaceticus, Burkholderia cepacia, Methylobacterium sp., Mycobacterium mucogenicum, Sphingomonas capsulata and Staphylococcus sp.). Adhesion was assessed by two distinct methods: thermodynamic prediction of adhesion potential by quantifying hydrophobicity\\u000a and the free energy of adhesion; and by microtiter plate assays. Biofilms
L�cia Chaves Sim�es; Manuel Sim�es; Maria Jo�o Vieira
Sugar fatty acid esters inhibit biofilm formation by food-borne pathogenic bacteria.
Effects of food additives on biofilm formation by food-borne pathogenic bacteria were investigated. Thirty-three potential food additives and 3 related compounds were added to the culture medium at concentrations from 0.001 to 0.1% (w/w), followed by inoculation and cultivation of five biofilm-forming bacterial strains for the evaluation of biofilm formation. Among the tested food additives, 21 showed inhibitory effects of biofilm formation by Staphylococcus aureus and Escherichia coli, and in particular, sugar fatty acid esters showed significant anti-biofilm activity. Sugar fatty acid esters with long chain fatty acid residues (C14-16) exerted their inhibitory effect at the concentration of 0.001% (w/w), but bacterial growth was not affected at this low concentration. Activities of the sugar fatty acid esters positively correlated with the increase of the chain length of the fatty acid residues. Sugar fatty acid esters inhibited the initial attachment of the S. aureus cells to the abiotic surface. Sugar fatty acid esters with long chain fatty acid residues (C14-16) also inhibited biofilm formation by Streptococcus mutans and Listeria monocytogenes at 0.01% (w/w), while the inhibition of biofilm formation by Pseudomonas aeruginosa required the addition of a far higher concentration (0.1% (w/w)) of the sugar fatty acid esters. PMID:20089325
Furukawa, Soichi; Akiyoshi, Yuko; O'Toole, George A; Ogihara, Hirokazu; Morinaga, Yasushi
Effect of estradiol on planktonic growth, coaggregation, and biofilm formation of the Prevotella intermedia group bacteria.
Alterations in the quantity and quality of biofilms at gingival margin are considered to play a role in the initiation and development of pregnancy-related gingivitis. Prevotella intermedia sensu lato is able to consume estradiol, the major sex hormone secreted during pregnancy, in the absence of vitamin K. The aim of the study was to examine the effect of estradiol on the planktonic growth, coaggregation, polysaccharide production, and biofilm formation of the P.�intermedia group bacteria, namely P.�intermedia, Prevotella nigrescens, and Prevotella pallens. In all experiments, the type strain (ATCC) and a clinical strain (AHN) of P.�intermedia, P.�nigrescens, and P.�pallens were incubated with the concentrations of 0, 30, 90, and 120�nmol/L of estradiol. Planktonic growth was assessed by means of the colony forming unit method, while coaggregation and biofilm formation were assessed by spectrophotometric methods. In the determination of protein and polysaccharide levels, the Bradford and phenol-sulfuric acid methods were used, respectively. P.�pallens AHN 9283 and P.�nigrescens ATCC 33563 increased their numbers at planktonic stage with increasing estradiol concentrations. In 48-h biofilm tests, elevated protein levels were found for both strains of P.�intermedia, and the strains P.�nigrescens ATCC 33563 and P.�pallens AHN 9283 in the presence of estradiol. The P.�intermedia strains also increased the levels of polysaccharide formation in the biofilm. Coaggregation of the P.�intermedia group organisms with Fusobacterium nucleatum was enhanced only in P.�intermedia AHN 8290. In conclusion, our in�vitro experiments indicate that estradiol regulates planktonic growth, coaggregation, polysaccharide production, and biofilm formation characteristics of P.�intermedia, P.�nigrescens, and P.�pallens differently. These results may, at least partly, explain the differences seen in their contribution to the pathogenesis of pregnancy-related gingivitis. PMID:24594108
Fteita, Dareen; K�n�nen, Eija; S�derling, Eva; G�rsoy, Ulvi Kahraman
Identification of pink-pigmented bacteria isolated from environmental water samples and their biofilm formation abilities.
Sixty-seven strains of pink-pigmented bacteria, which were isolated from environmental water samples collected nationwide, were identified by partial 16S rDNA sequence analysis. In addition, the biofilm formation ability of the isolates was experimentally investigated. We could identify only 2 strains at the species level: Pedobacter roseus HS-38 and Runella slithyformis HS-77. The results showed that of the strains tested, 22 strains (32.8%) were Pedobacter spp., which was most frequently identified, followed by 19 strains (28.4%) of Arcicella spp., 16 strains (23.9%) of Deinococcus spp., 5 strains (7.5%) of Roseomonas spp., 4 strains (6.0%) of Flectobacillus spp. and 1 strain (1.5%) of Runella sp. Most isolates showed low similarity values to previously known species, and they were found to be novel species. At a result, it was difficult to identify environmental water-derived pink-pigmented bacteria at the species level. On the other hand, when we measured the absorbance by the crystal violet staining to examine the quantities of biofilm formation of these strains, fifty-five (82.0%) of the 67 isolates formed biofilm. The absorbance of Deinococcus sp. HS-75 was the highest (3.56). When comparing the absorbance values among the genera, Roseomonas spp. showed the highest absorbance (mean:1.62), followed by Deinococcus spp. (mean: 1.03), and Arcicella spp. (mean: 1.01). Strains of Flectobacillus spp. (mean: 0.48) and Pedobacter spp. (mean: 0.42) showed lower absorbance values. As above, it was shown that, at the species level, the pink-pigmented bacteria in the water in the Japanese environment had various levels of ability to form biofilm. PMID:18661678
Furuhata, Katsunori; Kato, Yuko; Goto, Keiichi; Saitou, Keiko; Sugiyama, Jun-Ichi; Hara, Motonobu; Fukuyama, Masahumi
Inactivation of biofilm bacteria.
The current project was developed to examine inactivation of biofilm bacteria and to characterize the interaction of biocides with pipe surfaces. Unattached bacteria were quite susceptible to the variety of disinfectants tested. Viable bacterial counts were reduced 99% by exposure to 0.08 mg of hypochlorous acid (pH 7.0) per liter (1 to 2 degrees C) for 1 min. For monochloramine, 94 mg/liter was required to kill 99% of the bacteria within 1 min. These results were consistent with those found by other investigators. Biofilm bacteria grown on the surfaces of granular activated carbon particles, metal coupons, or glass microscope slides were 150 to more than 3,000 times more resistant to hypochlorous acid (free chlorine, pH 7.0) than were unattached cells. In contrast, resistance of biofilm bacteria to monochloramine disinfection ranged from 2- to 100-fold more than that of unattached cells. The results suggested that, relative to inactivation of unattached bacteria, monochloramine was better able to penetrate and kill biofilm bacteria than free chlorine. For free chlorine, the data indicated that transport of the disinfectant into the biofilm was a major rate-limiting factor. Because of this phenomenon, increasing the level of free chlorine did not increase disinfection efficiency. Experiments where equal weights of disinfectants were used suggested that the greater penetrating power of monochloramine compensated for its limited disinfection activity. These studies showed that monochloramine was as effective as free chlorine for inactivation of biofilm bacteria. The research provides important insights into strategies for control of biofilm bacteria. Images
LeChevallier, M W; Cawthon, C D; Lee, R G
There is a paucity of information concerning any link between the microorganisms commonly found in biofilms of drinking water systems and their impacts on human health. For bacteria, culture-based techniques detect only a limited number of the total microorganisms associated wit...
Effect of temperature on biofilm formation by Antarctic marine bacteria in a microfluidic device.
Polar biofilms have become an increasingly popular biological issue because new materials and phenotypes have been discovered in microorganisms in the polar region. Various environmental factors affect the functionality and adaptation of microorganisms. Because the polar region represents an extremely cold environment, polar microorganisms have a functionality different from that of normal microorganisms. Thus, determining the effective temperature for the development of polar biofilms is crucial. Here, we present a simple, novel one-pot assay for analysis of the effect of temperature on formation of Antarctic bacterial biofilm using a microfluidic system where continuous temperature gradients are generated. We find that a specific range of temperature is required for the growth of biofilms. Thus, this microfluidic approach provides precise information regarding the effective temperature for polar biofilm development with a new high-throughput screening format. PMID:24513116
Jeong, Heon-Ho; Jeong, Seong-Geun; Park, Aeri; Jang, Sung-Chan; Hong, Soon Gyu; Lee, Chang-Soo
Screening, characterization and biofilm formation of nickel resistant bacteria isolated from indigenous environment.
Nickel resistant bacteria (ZB, ZC, ZD, ZL, ZK and S1X) were isolated from industrial effluents and corroded iron pieces from indigenous environment of Punjab, Pakistan. These six strains could tolerate nickel at different levels with ZB, ZC, ZD, ZL, ZK, and S1X having 233, 225, 267, 233, 228 and 296 mM minimum inhibitory concentration (MIC) of nickel ions, respectively. These bacteria were sensitive to Cu(+2), Cr(+3), Co(+2), and Al(+3) as they did not grow even in the presence of 1 mM concentration of all these ions in minimal medium, whereas all of them were resistant to Fe3 upto 1.3 mM in minimal medium. The best appropriate temperature for nickel resistant bacteria was 37 degrees C and all of them showed maximum growth at pH 8. These bacteria were characterized morphologically and biochemically. Biofilm forming ability of the bacteria was checked with and without nickel stress and it was found that strains ZK and S1X were able to form a compact biofilm even under nickel stress. The sequencing of 16S rRNA-encoding genes from these nickel resistant bacteria showed that they belonged to four different genera namely, Klebsiella, Pseudomonas, Bacillus and Cronobacter. PMID:24730136
Wadood, Hafiz Z; Sabri, Anjum N
Engineering biofilm formation and dispersal
Anywhere water is in the liquid state, bacteria will exist as biofilms, which are complex communities of cells cemented together. Although frequently associated with disease and biofouling, biofilms are also important for engineering applications, such as bioremediation, biocatalysis and microbial fuel cells. Here we review approaches to alter genetic circuits and cell signaling toward controlling biofilm formation, and emphasize utilizing these tools for engineering applications. Based on a better understanding of the genetic basis of biofilm formation, we find that biofilms may be controlled by manipulating extracellular signals and that they may be dispersed using conserved intracellular signals and regulators. Biofilms could also be formed at specific locations where they might be engineered to make chemicals or treat human disease.
Wood, Thomas K.; Hong, Seok Hoon; Ma, Qun
Calcium carbonate precipitation by heterotrophic bacteria isolated from biofilms formed on deteriorated ignimbrite stones: influence of calcium on EPS production and biofilm formation by these isolates.
Heterotrophic CaCO3-precipitating bacteria were isolated from biofilms on deteriorated ignimbrites, siliceous acidic rocks, from Morelia Cathedral (Mexico) and identified as Enterobacter cancerogenus (22e), Bacillus sp. (32a) and Bacillus subtilis (52g). In solid medium, 22e and 32a precipitated calcite and vaterite while 52g produced calcite. Urease activity was detected in these isolates and CaCO3 precipitation increased in the presence of urea in the liquid medium. In the presence of calcium, EPS production decreased in 22e and 32a and increased in 52g. Under laboratory conditions, ignimbrite colonization by these isolates only occurred in the presence of calcium and no CaCO3 was precipitated. Calcium may therefore be important for biofilm formation on stones. The importance of the type of stone, here a siliceous stone, on biological colonization is emphasized. This calcium effect has not been reported on calcareous materials. The importance of the effect of calcium on EPS production and biofilm formation is discussed in relation to other applications of CaCO3 precipitation by bacteria. PMID:24689777
L�pez-Moreno, Ang�lica; Sep�lveda-S�nchez, Jos� David; Mercedes Alonso Guzm�n, Elia Mercedes; Le Borgne, Sylvie
Effect of Algae and Plant Lectins on Planktonic Growth and Biofilm Formation in Clinically Relevant Bacteria and Yeasts
This study aimed to evaluate the abilities of plant and algae lectins to inhibit planktonic growth and biofilm formation in bacteria and yeasts. Initially, ten lectins were tested on Staphylococcus epidermidis, Staphylococcus aureus, Klebsiella oxytoca, Pseudomonas aeruginosa, Candida albicans, and C. tropicalis at concentrations of 31.25 to 250??g/mL. The lectins from Cratylia floribunda (CFL), Vatairea macrocarpa (VML), Bauhinia bauhinioides (BBL), Bryothamnion seaforthii (BSL), and Hypnea musciformis (HML) showed activities against at least one microorganism. Biofilm formation in the presence of the lectins was also evaluated; after 24?h of incubation with the lectins, the biofilms were analyzed by quantifying the biomass (by crystal violet staining) and by enumerating the viable cells (colony-forming units). The lectins reduced the biofilm biomass and/or the number of viable cells to differing degrees depending on the microorganism tested, demonstrating the different characteristics of the lectins. These findings indicate that the lectins tested in this study may be natural alternative antimicrobial agents; however, further studies are required to better elucidate the functional use of these proteins.
Vasconcelos, Mayron Alves; Arruda, Francisco Vassiliepe Sousa; Carneiro, Victor Alves; Silva, Helton Colares; Nascimento, Kyria Santiago; Sampaio, Alexandre Holanda; Cavada, Benildo; Teixeira, Edson Holanda; Henriques, Mariana
Isolation and characterization of acyl homoserine lactone-producing bacteria during an urban river biofilm formation.
The presence and diversity of acyl homoserine lactone (AHL)-producers in an urban river biofilm were investigated during 60-day biofilm formation. AHL biosensors detected the presence of AHL-producers in 1-60-day river biofilms. Screening for AHL-producers resulted in 17 Aeromonas spp., 3 Pseudomonas spp., 3 Ensifer spp., and 1 Acinetobacter sp. Among these isolates, six of them were closely related to Acinetobacter tjernbergiae, Aeromonas allosaccharophila, Aeromonas aquariorum, Aeromonas jandaei, Pseudomonas panipatensis, and Ensifer adhaerens and represented novel AHL-producing species. Thin layer chromatography revealed that C4-homoserine lactone was prevailing in Aeromonas spp., whereas C6- and C8-homoserine lactones and their derivatives were prevailing in other strains. Using degenerate primers, novel AHL synthetase genes from the three Ensifer spp. were successfully amplified. This study reports for the first time the diversity of AHL-producers from a river biofilm and the variety of novel AHL synthetase genes in Ensifer group. PMID:23090571
Huang, Yili; Zhang, Jing; Yu, Zhiliang; Zeng, Yanhua; Chen, Yuqi
Molecular mechanisms of Staphylococcus epidermidis biofilm formation
Coagulase-negative staphylococci, mainly Staphylococcus epidermidis, are the predominant cause of implanted medical-device related infections. The formation of adherent multilayered biofilms embedded into a glycocalyx composed of exopolysaccharides on implanted devices is believed to be essential for the pathogenesis of S. epidermidis infections. Biofilm formation may be separated into primary attachment of bacteria to native or modified polymer surfaces followed by
D. Mack
The ability to form biofilms is a universal attribute of bacteria. Biofilms are multicellular communities held together by a self-produced extracellular matrix. The mechanisms that different bacteria employ to form biofilms vary, frequently depending on environmental conditions and specific strain attributes. In this review, we emphasize four well-studied model systems to give an overview of how several organisms form biofilms: Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus. Using these bacteria as examples, we discuss the key features of biofilms as well as mechanisms by which extracellular signals trigger biofilm formation. PMID:20519345
L�pez, Daniel; Vlamakis, Hera; Kolter, Roberto
Biofilm formation in a hot water system
The biofilm formation rate was measured in situ in a hot water system in an apartment building by specially designed sampling equipment, and the net growth of the suspended bacteria was measured by incubation of water samples with the indigeneous bacteria. The biofilm formation rate reached a higher level in the hot water distribution system (2.1 d-1 to 2.3 d-1)
L. K. Bagh; H.-J. Albrechtsen; E. Arvin; K. Ovesen
Inactivation ofBiofilm Bacteria
Thecurrent project was developed toexamine inactivation ofbiofilm bacteria andtocharacterize the interaction ofbiocides withpipesurfaces. Unattached bacteria were quite susceptible tothevariety of disinfectants tested. Viable bacterial counts were reduced 99%byexposureto0.08 mg ofhypochlorous acid (pH7.0) perliter (1to2�C) for1min.Formonochloramine, 94mg\\/liter wasrequired tokill 99%ofthebacteria within 1min.Theseresults wereconsistent withthose found byother investigators. Biofilm bacteria grown on thesurfaces ofgranular activated carbon particles, metal coupons,orglass microscope slides were
Antibacterial effects of silver nanoparticles on gram-negative bacteria: influence on the growth and biofilms formation, mechanisms of action.
Antibacterial action of silver nanoparticles (AgNP) on Gram-negative bacteria (planctonic cells and biofilms) is reported in this study. AgNP of 8.3 nm in diameter stabilized by hydrolyzed casein peptides strongly inhibited biofilms formation of Escherichia coli AB1157, Pseudomonas aeruginosa PAO1 and Serratia proteamaculans 94 in concentrations of 4-5 ?g/ml, 10 ?g/ml and 10-20 ?g/ml, respectively. The viability of E. coli AB1157 cells in biofilms was considerably reduced by AgNP concentrations above 100 to -150 ?g/ml. E. coli strains with mutations in genes responsible for the repair of DNA containing oxidative lesions (mutY, mutS, mutM, mutT, nth) were less resistant to AgNP than wild type strains. This suggests that these genes may be involved in the repair of DNA damage caused by AgNP. E. coli mutants deficient in excision repair, SOS-response and in the synthesis of global regulators RpoS, CRP protein and Lon protease present similar resistance to AgNP as wild type cells. LuxI/LuxR Quorum Sensing systems did not participate in the control of sensitivity to AgNP of Pseudomonas and Serratia. E. coli mutant strains deficient in OmpF or OmpC porins were 4-8 times more resistant to AgNP as compared to the wild type strain. This suggests that porins have an important function related AgNP antibacterial effects. PMID:23006569
Radzig, M A; Nadtochenko, V A; Koksharova, O A; Kiwi, J; Lipasova, V A; Khmel, I A
Biofilm formation by haloarchaea.
A fluorescence-based live-cell adhesion assay was used to examine biofilm formation by 20 different haloarchaea, including species of Halobacterium, Haloferax and Halorubrum, as well as novel natural isolates from an Antarctic salt lake. Thirteen of the 20 tested strains significantly adhered (P-value ?< 0.05) to a plastic surface. Examination of adherent cell layers on glass surfaces by differential interference contrast, fluorescence and confocal microscopy showed two types of biofilm structures. Carpet-like, multi-layered biofilms containing micro- and macrocolonies (up to 50 ?m in height) were formed by strains of Halobacterium salinarum and the Antarctic isolate t-ADL strain DL24. The second type of biofilm, characterized by large aggregates of cells adhering to surfaces, was formed by Haloferax volcanii DSM 3757T and Halorubrum lacusprofundi DL28. Staining of the biofilms formed by the strongly adhesive haloarchaeal strains revealed the presence of extracellular polymers, such as eDNA and glycoconjugates, substances previously shown to stabilize bacterial biofilms. For Hbt. salinarum DSM 3754T and Hfx. volcanii DSM 3757T , cells adhered within 1 day of culture and remained viable for at least 2 months in mature biofilms. Adherent cells of Hbt. salinarum DSM 3754T showed several types of cellular appendages that could be involved in the initial attachment. Our results show that biofilm formation occurs in a surprisingly wide variety of haloarchaeal species. PMID:23057712
Fr�ls, Sabrina; Dyall-Smith, Mike; Pfeifer, Felicitas
Biofilm-growing intestinal anaerobic bacteria.
Sessile growth of anaerobic bacteria from the human intestinal tract has been poorly investigated, so far. We recently reported data on the close association existing between biliary stent clogging and polymicrobial biofilm development in its lumen. By exploiting the explanted stents as a rich source of anaerobic bacterial strains belonging to the genera Bacteroides, Clostridium, Fusobacterium, Finegoldia, Prevotella, and Veillonella, the present study focused on their ability to adhere, to grow in sessile mode and to form in vitro mono- or dual-species biofilms. Experiments on dual-species biofilm formation were planned on the basis of the anaerobic strains isolated from each clogged biliary stent, by selecting those in which a couple of anaerobic strains belonging to different species contributed to the polymicrobial biofilm development. Then, strains were investigated by field emission scanning electron microscopy and confocal laser scanning microscopy to reveal if they are able to grow as mono- and/or dual-species biofilms. As far as we know, this is the first report on the ability to adhere and form mono/dual-species biofilms exhibited by strains belonging to the species Bacteroides oralis, Clostridium difficile, Clostridium baratii, Clostridium fallax, Clostridium bifermentans, Finegoldia magna, and Fusobacterium necrophorum. PMID:22444687
Donelli, Gianfranco; Vuotto, Claudia; Cardines, Rita; Mastrantonio, Paola
Emergent Bacteria in Cystic Fibrosis: In Vitro Biofilm Formation and Resilience under Variable Oxygen Conditions
Concurrent to conventional bacterial pathogens, unusual microbes are emerging from cystic fibrosis (CF) airways. Nonetheless, little is known about the contribution of these newly microbes to the resilience of CF-associated biofilms, particularly under variable-oxygen concentrations that are known to occur in vivo in the mucus of CF patients. Two CF-emergent bacterial species, Inquilinus limosus and Dolosigranulum pigrum, and the major pathogen Pseudomonas aeruginosa were studied in terms of biofilm development and antibiotic susceptibilities under in vitro atmospheres with different oxygen availabilities. All species were able to develop in vitro biofilms under different oxygen-available environments, with D. pigrum accumulating high amounts of biomass and respiratory activities. When established, biofilms were of difficult eradication, with antibiotics losing their effectiveness in comparison with the corresponding planktonic populations. Surprisingly, biofilms of each emergent organism displayed multidrug resistance under aerobic environments, enduring even in low-oxygen atmospheres. This study suggests a potential prospect on the impact of nonconventional organisms I. limosus and D. pigrum on CF lung infections, demonstrating capacity to adapt to biofilm mode of life under restricted-oxygen atmospheres resembling CF airways, which may ultimately endanger the efficacy of currently used antibiotic regimens.
Lopes, Susana P.; Azevedo, Nuno F.; Pereira, Maria O.
Released exopolysaccharide (r-EPS) produced from probiotic bacteria reduce biofilm formation of enterohemorrhagic Escherichia coli O157:H7.
Here, we characterized released-exopolysaccharides (r-EPS) from Lactobacillus acidophilus A4 with the goal of identifying natural compounds that represses biofilm formation. In plastic 96-well microplates that contained 1.0 mg/ml of r-EPS, enterohemorrhagic Escherichia coli (EHEC) biofilms were dramatically decreased by 87% and 94% on polystyrene and polyvinyl chloride (PVC) surfaces, respectively. In the presence of r-EPS, neither their growth rate nor their autoinducer-2-like activity was affected on the EHEC O157:H7. Importantly, consistent reduction in biofilm formation was also observed when r-EPS was applied to the continuous-flow chamber models. In addition, we found that adding r-EPS significantly repressed biofilm formation by affecting genes related to curli production (crl, csgA, and csgB) and chemotaxis (cheY) in transcriptome analysis. Furthermore, these r-EPS could prevent biofilm formation by a wide range of Gram-negative and -positive pathogens. This property may lead to the development of novel food-grade adjuncts for microbial biofilm control. PMID:19103165
Kim, Younghoon; Oh, Sejong; Kim, Sae Hun
Antibiotic susceptibility of coagulase-negative staphylococci isolated from very low birth weight babies: comprehensive comparisons of bacteria at different stages of biofilm formation
Background Coagulase-negative staphylococci are major causes of bloodstream infections in very low birth weight babies cared for in Neonatal Intensive Care Units. The virulence of these bacteria is mainly due to their ability to form biofilms on indwelling medical devices. Biofilm-related infections often fail to respond to antibiotic chemotherapy guided by conventional antibiotic susceptibility tests. Methods Coagulase-negative staphylococcal blood culture isolates were grown in different phases relevant to biofilm formation: planktonic cells at mid-log phase, planktonic cells at stationary phase, adherent monolayers and mature biofilms and their susceptibilities to conventional antibiotics were assessed. The effects of oxacillin, gentamicin, and vancomycin on preformed biofilms, at the highest achievable serum concentrations were examined. Epifluorescence microscopy and confocal laser scanning microscopy in combination with bacterial viability staining and polysaccharide staining were used to confirm the stimulatory effects of antibiotics on biofilms. Results Most coagulase-negative staphylococcal clinical isolates were resistant to penicillin G (100%), gentamicin (83.3%) and oxacillin (91.7%) and susceptible to vancomycin (100%), ciprofloxacin (100%), and rifampicin (79.2%). Bacteria grown as adherent monolayers showed similar susceptibilities to their planktonic counterparts at mid-log phase. Isolates in a biofilm growth mode were more resistant to antibiotics than both planktonic cultures at mid-log phase and adherent monolayers; however they were equally resistant or less resistant than planktonic cells at stationary phase. Moreover, for some cell-wall active antibiotics, concentrations higher than conventional MICs were required to prevent the establishment of planktonic cultures from biofilms. Finally, the biofilm-growth of two S. capitis isolates could be enhanced by oxacillin at the highest achievable serum concentration. Conclusion We conclude that the resistance of coagulase-negative staphylococci to multiple antibiotics initially remain similar when the bacteria shift from a planktonic growth mode into an early attached mode, then increase significantly as the adherent mode further develops. Furthermore, preformed biofilms of some CoNS are enhanced by oxacillin in a dose-dependent manner.
Regulation of flagellar motility during biofilm formation.
Many bacteria swim in liquid or swarm over solid surfaces by synthesizing rotary flagella. The same bacteria that are motile also commonly form nonmotile multicellular aggregates called biofilms. Biofilms are an important part of the lifestyle of pathogenic bacteria, and it is assumed that there is a motility-to-biofilm transition wherein the inhibition of motility promotes biofilm formation. The transition is largely inferred from regulatory mutants that reveal the opposite regulation of the two phenotypes. Here, we review the regulation of motility during biofilm formation in Bacillus, Pseudomonas, Vibrio, and Escherichia, and we conclude that the motility-to-biofilm transition, if necessary, likely involves two steps. In the short term, flagella are functionally regulated to either inhibit rotation or modulate the basal flagellar reversal frequency. Over the long term, flagellar gene transcription is inhibited and in the absence of de novo synthesis, flagella are diluted to extinction through growth. Both short-term and long-term motility inhibition is likely important to stabilize cell aggregates and optimize resource investment. We emphasize the newly discovered flagellar functional regulators and speculate that others await discovery in the context of biofilm formation. PMID:23480406
Guttenplan, Sarah B; Kearns, Daniel B
A new biofilm-associated colicin with increased efficiency against biofilm bacteria.
Formation of bacterial biofilm communities leads to profound physiological modifications and increased physical and metabolic exchanges between bacteria. It was previously shown that bioactive molecules produced within the biofilm environment contribute to bacterial interactions. Here we describe new pore-forming colicin R, specifically produced in biofilms formed by the natural isolate Escherichia coli ROAR029 but that cannot be detected under planktonic culture conditions. We demonstrate that an increased SOS stress response within mature biofilms induces SOS-dependent colicin R expression. We provide evidence that colicin R displays increased activity against E. coli strains that have a reduced lipopolysaccharide length, such as the pathogenic enteroaggregative E. coli LF82 clinical isolate, therefore pointing to lipopolysaccharide size as an important determinant for resistance to colicins. We show that colicin R toxicity toward E. coli LF82 is increased under biofilm conditions compared with planktonic susceptibility and that release of colicin R confers a strong competitive advantage in mixed biofilms by rapidly outcompeting sensitive neighboring bacteria. This work identifies the first biofilm-associated colicin that preferentially targets biofilm bacteria. Furthermore, it indicates that the study of antagonistic molecules produced in biofilm and multispecies contexts could reveal unsuspected, ecologically relevant bacterial interactions influencing population dynamics in natural environments. PMID:24451204
Rendueles, Olaya; Beloin, Christophe; Latour-Lambert, Patricia; Ghigo, Jean-Marc
Spaceflight promotes biofilm formation by Pseudomonas aeruginosa.
Understanding the effects of spaceflight on microbial communities is crucial for the success of long-term, manned space missions. Surface-associated bacterial communities, known as biofilms, were abundant on the Mir space station and continue to be a challenge on the International Space Station. The health and safety hazards linked to the development of biofilms are of particular concern due to the suppression of immune function observed during spaceflight. While planktonic cultures of microbes have indicated that spaceflight can lead to increases in growth and virulence, the effects of spaceflight on biofilm development and physiology remain unclear. To address this issue, Pseudomonas aeruginosa was cultured during two Space Shuttle Atlantis missions: STS-132 and STS-135, and the biofilms formed during spaceflight were characterized. Spaceflight was observed to increase the number of viable cells, biofilm biomass, and thickness relative to normal gravity controls. Moreover, the biofilms formed during spaceflight exhibited a column-and-canopy structure that has not been observed on Earth. The increase in the amount of biofilms and the formation of the novel architecture during spaceflight were observed to be independent of carbon source and phosphate concentrations in the media. However, flagella-driven motility was shown to be essential for the formation of this biofilm architecture during spaceflight. These findings represent the first evidence that spaceflight affects community-level behaviors of bacteria and highlight the importance of understanding how both harmful and beneficial human-microbe interactions may be altered during spaceflight. PMID:23658630
Kim, Wooseong; Tengra, Farah K; Young, Zachary; Shong, Jasmine; Marchand, Nicholas; Chan, Hon Kit; Pangule, Ravindra C; Parra, Macarena; Dordick, Jonathan S; Plawsky, Joel L; Collins, Cynthia H
Biofilm formation in bacterial pathogens of veterinary importance.
Bacterial biofilms are structured communities of bacterial cells enclosed in a self-produced polymer matrix that is attached to a surface. Biofilms protect and allow bacteria to survive and thrive in hostile environments. Bacteria within biofilms can withstand host immune responses, and are much less susceptible to antibiotics and disinfectants when compared with their planktonic counterparts. The ability to form biofilms is now considered a universal attribute of micro-organisms. Diseases associated with biofilms require novel methods for their prevention, diagnosis and treatment; this is largely due to the properties of biofilms. Surprisingly, biofilm formation by bacterial pathogens of veterinary importance has received relatively little attention. Here, we review the current knowledge of bacterial biofilms as well as studies performed on animal pathogens. PMID:20969814
Jacques, Mario; Aragon, Virginia; Tremblay, Yannick D N
Biofilm formation in water cooling systems
Biofilm formation on stainless steel samples immersed in cooling water has been evaluated by exposing metal samples to cooling seawater for 30 days. Anaerobic bacteria were then at 1.6 � 106\\/cm2, with sulphate-reducing species predominating. Aerobic bacteria and fungi were 2600 and 140\\/cm2, respectively. After 60 days, numbers of aerobic microorganisms remained constant whereas the count of anaerobic microorganisms had
M. T. S. Lutterbach; F. P. Fran�a
Biofilm formation in a hot water system.
The biofilm formation rate was measured in situ in a hot water system in an apartment building by specially designed sampling equipment, and the net growth of the suspended bacteria was measured by incubation of water samples with the indigeneous bacteria. The biofilm formation rate reached a higher level in the hot water distribution system (2.1 d(-1) to 2.3 d(-1)) than in the hot water tank (1.4 d(-1) to 2.2 d(-1)) indicating an important area for surface associated growth. The net growth rate of the suspended bacteria measured in hot water from the top, middle and bottom of the hot water tank, in the sludge, or in the water from the distribution system was negligible. This indicated that bacterial growth took place on the inner surfaces in the hot water system and biofilm formation and detachment of bacteria could account for most of the suspended bacteria actually measured in hot water. Therefore, attempts to reduce the number of bacteria in a hot water system have to include the distribution system as well as the hot water tank. PMID:12448457
Bagh, L K; Albrechtsen, H J; Arvin, E; Ovesen, K
Biofilm formation on oral piercings
Summary��PURPOSE: Biofilms on oral piercings might serve as a bacterial reservoir in the host and lead to bacteraemia and even septic\\u000a complications. The use of piercing materials less susceptible to biofilm accumulation could contribute to alleviation of problems.\\u000a The present study aimed to assess biofilm formation on four commercially available, surface characterized piercing materials\\u000a in vitro (polytetrafluoroethylene, titanium, stainless steel,
Ines Kapferer; Christoph Steiner; Ulrike Beier; Natalia Schiefermeier; Markus Nagl; Frederik Klauser
This project addressed four major areas of investigation: i) characterization of formation of Cellulomonas uda biofilms on cellulose; ii) characterization of Clostridium phytofermentans biofilm development; colonization of cellulose and its regulation; iii) characterization of Thermobifida fusca biofilm development; colonization of cellulose and its regulation; and iii) description of the architecture of mature C. uda, C. phytofermentans, and T. fusca biofilms. This research is aimed at advancing understanding of biofilm formation and other complex processes involved in the degradation of the abundant cellulosic biomass, and the biology of the microbes involved. Information obtained from these studies is invaluable in the development of practical applications, such as the single-step bioconversion of cellulose-containing residues to fuels and other bioproducts. Our results have clearly shown that cellulose-decomposing microbes rapidly colonize cellulose and form complex structures typical of biofilms. Furthermore, our observations suggest that, as cells multiply on nutritive surfaces during biofilms formation, dramatic cell morphological changes occur. We speculated that morphological changes, which involve a transition from rod-shaped cells to more rounded forms, might be more apparent in a filamentous microbe. In order to test this hypothesis, we included in our research a study of biofilm formation by T. fusca, a thermophilic cellulolytic actinomycete commonly found in compost. The cellulase system of T. fusca has been extensively detailed through the work of David Wilson and colleagues at Cornell, and also, genome sequence of a T. fusca strain has been determine by the DOE Joint Genome Institute. Thus, T. fusca is an excellent subject for studies of biofilm development and its potential impacts on cellulose degradation. We also completed a study of the chitinase system of C. uda. This work provided essential background information for understanding how C. uda colonizes and degrades insoluble substrates. Major accomplishments of the project include: � Development of media containing dialysis tubing (described by the manufacturer as �regenerated cellulose�) as sole carbon and energy source and a nutritive surface for the growth of cellulolytic bacteria, and development of various microscopic methods to image biofilms on dialysis tubing. � Demonstration that cultures of C. phytofermentans, an obligate anaerobe, C. uda, a facultative aerobe, and T. fusca, a filamentous aerobe, formed microbial communities on the surface of dialysis tubing, which possessed architectural features and functional characteristics typical of biofilms. � Demonstration that biofilm formation on the nutritive surface, cellulose, involves a complex developmental processes, including colonization of dialysis tubing, formation of cell clusters attached to the nutritive surface, cell morphological changes, formation of complex structures embedded in extracellular polymeric matrices, and dispersal of biofilm communities as the nutritive surface is degraded. � Determination of surface specificity and regulatory aspects of biofilm formation by C. phytofermentans, C. uda, and T. fusca. � Demonstration that biofilm formation by T. fusca forms an integral part of the life cycle of this filamentous cellulolytic bacterium, including studies on the role of mycelial pellet formation in the T. fusca life cycle and a comparison of mycelial pellets to surface-attached T. fusca biofilms. � Characterization of T. fusca biofilm EPS, including demonstration of a functional role for EPS constituents. � Correlation of T. fusca developmental life cycle and cellulase gene expression.
Leschine, Susan
Biofilm Formation by Candida dubliniensis
Candida dubliniensis is an opportunistic yeast closely related to Candida albicans that has been recently implicated in oropharyngeal candidiasis in human immunodeficiency virus-infected patients. Most manifes- tations of candidiasis are associated with biofilm formation, with cells in biofilms displaying properties dramatically different from free-living cells grown under normal laboratory conditions. Here, we report on the development of in vitro models
Tryptophan Inhibits Biofilm Formation by Pseudomonas aeruginosa
Biofilm formation by Pseudomonas aeruginosa has been implicated in the pathology of chronic wounds. Both the d and l isoforms of tryptophan inhibited P. aeruginosa biofilm formation on tissue culture plates, with an equimolar ratio of d and l isoforms producing the greatest inhibitory effect. Addition of d-/l-tryptophan to existing biofilms inhibited further biofilm growth and caused partial biofilm disassembly. Tryptophan significantly increased swimming motility, which may be responsible in part for diminished biofilm formation by P. aeruginosa.
Brandenburg, Kenneth S.; Rodriguez, Karien J.; McAnulty, Jonathan F.; Murphy, Christopher J.; Abbott, Nicholas L.; Schurr, Michael J.
Biofilm formation of Pasteurella multocida on bentonite clay
Background and objectives Biofilms are structural communities of bacterial cells enshrined in a self produced polymeric matrix. The studies on biofilm formation of Pasteurella multocida have become imperative since it is a respiratory pathogen and its biofilm mode could possibly be one of its virulence factors for survival inside a host. The present study describes a biofilm assay for P. multocida on inert hydrophilic material called bentonite clay. Materials and methods The potential of the organism to form in vitro biofilm was assessed by growing the organism under nutrient restriction along with the inert substrate bentonite clay, which will provide a surface for attachment. For quantification of biofilm, plate count by the spread plate method was employed. Capsule production of the attached bacteria was demonstrated by light microscopic examination following Maneval staining and capsular polysaccharide estimation was done using standard procedures. Results and Conclusion The biofilm formation peaked on the third day of incubation (1.54 �106 cfu/g of bentonite clay) while the planktonic cells were found to be at a maximum on day one post inoculation (8.10 �108 cfu/ml of the broth). Maneval staining of late logarithmic phase biofilm cultures revealed large aggregates of bacterial cells, bacteria appearing as chains or as a meshwork. The capsular polysaccharide estimation of biofilm cells revealed a 3.25 times increase over the planktonic bacteria. The biofilm cells cultured on solid media also produced some exclusive colony morphotypes.
Rajagopal, Ramachandranpillai; Nair, Govindapillai Krishnan; Mini, Mangattumuruppel; Joseph, Leo; Saseendranath, Mapranath Raghavan; John, Koshy
Biomolecular mechanisms of staphylococcal biofilm formation.
The multitude of biomolecular and regulatory factors involved in staphylococcal adhesion and biofilm formation owe much to their ability to colonize surfaces, allowing the biofilm form to become the preferential bacterial phenotype. Judging by total number, biomass and variety of environments colonized, bacteria can be categorized as the most successful lifeform on earth. This is due to the ability of bacteria and other microorganisms to respond phenotypically via biomolecular processes to the stresses of their surrounding environment. This review focuses on the specific pathways involved in the adhesion of the Gram-positive bacteria Staphylococcus epidermidis and Staphylococcus aureus with reference to the role of specific cell surface adhesins, the ica operon, accumulation-associated proteins and quorum-sensing systems and their significance in medical device-related infection. PMID:23534362
Laverty, Garry; Gorman, Sean P; Gilmore, Brendan F
Ginger Extract Inhibits Biofilm Formation by Pseudomonas aeruginosa PA14
Bacterial biofilm formation can cause serious problems in clinical and industrial settings, which drives the development or screening of biofilm inhibitors. Some biofilm inhibitors have been screened from natural products or modified from natural compounds. Ginger has been used as a medicinal herb to treat infectious diseases for thousands of years, which leads to the hypothesis that it may contain chemicals inhibiting biofilm formation. To test this hypothesis, we evaluated ginger�s ability to inhibit Pseudomonas aeruginosa PA14 biofilm formation. A static biofilm assay demonstrated that biofilm development was reduced by 39�56% when ginger extract was added to the culture. In addition, various phenotypes were altered after ginger addition of PA14. Ginger extract decreased production of extracellular polymeric substances. This finding was confirmed by chemical analysis and confocal laser scanning microscopy. Furthermore, ginger extract formed noticeably less rugose colonies on agar plates containing Congo red and facilitated swarming motility on soft agar plates. The inhibition of biofilm formation and the altered phenotypes appear to be linked to a reduced level of a second messenger, bis-(3?-5?)-cyclic dimeric guanosine monophosphate. Importantly, ginger extract inhibited biofilm formation in both Gram-positive and Gram-negative bacteria. Also, surface biofilm cells formed with ginger extract detached more easily with surfactant than did those without ginger extract. Taken together, these findings provide a foundation for the possible discovery of a broad spectrum biofilm inhibitor.
Kim, Han-Shin; Park, Hee-Deung
Small RNAs and their role in biofilm formation
The formation of biofilms is initiated by bacteria transitioning from the planktonic to the surface associated mode of growth. Several regulatory systems have been described to govern the initiation and subsequent formation of biofilms. Recent evidence suggests that regulatory networks governing the decision of bacteria whether to attach and form biofilms or remain as planktonic cells, are further subject to regulation by small non-coding RNAs (sRNAs). This is accomplished by sRNAs finetuning regulatory networks enabling concentration-specific responses, by sequestering, antagonizing or activating regulatory proteins in response to environmental cues, or by directly affecting the synthesis of proteins promoting/disfavoring the formation of biofilms. This review is aimed at giving an overview of the contribution of sRNAs in regulating the switch from the planktonic to the sessile bacterial lifestyle by highlighting how sRNAs converge with known regulatory systems required for biofilm formation.
Chambers, Jacob R.; Sauer, Karin
Shewanella putrefaciens Adhesion and Biofilm Formation on Food Processing Surfaces
Laboratory model systems were developed for studying Shewanella putrefaciens adhesion and biofilm formation under batch and flow conditions. S. putrefaciens plays a major role in food spoilage and may cause microbially induced corrosion on steel surfaces. S. putrefaciens bacteria suspended in buffer adhered readily to stainless steel surfaces. Maximum numbers of adherent bacteria per square centimeter were reached in 8 h at 25�C and reflected the cell density in suspension. Numbers of adhering bacteria from a suspension containing 108 CFU/ml were much lower in a laminar flow system (modified Robbins device) (reaching 102 CFU/cm2) than in a batch system (reaching 107 CFU/cm2), and maximum numbers were reached after 24 h. When nutrients were supplied, S. putrefaciens grew in biofilms with layers of bacteria. The rate of biofilm formation and the thickness of the film were not dependent on the availability of carbohydrate (lactate or glucose) or on iron starvation. The number of S. putrefaciens bacteria on the surface was partly influenced by the presence of other bacteria (Pseudomonas fluorescens) which reduced the numbers of S. putrefaciens bacteria in the biofilm. Numbers of bacteria on the surface must be quantified to evaluate the influence of environmental factors on adhesion and biofilm formation. We used a combination of fluorescence microscopy (4?,6?-diamidino-2-phenylindole staining and in situ hybridization, for mixed-culture studies), ultrasonic removal of bacteria from surfaces, and indirect conductometry and found this combination sufficient to quantify bacteria on surfaces.
Bagge, Dorthe; Hjelm, Mette; Johansen, Charlotte; Huber, Ingrid; Gram, Lone
Yeasts and lactic acid bacteria mixed-specie biofilm formation is a promising cell immobilization technology for ethanol fermentation.
We previously found that some Saccharomyces cerevisiae and Lactobacillus plantarum remarkably formed mixed-specie biofilm in a static co-culture and deduced that this biofilm had potential as immobilized cells. We investigated the application of mixed-specie biofilm formed by S. cerevisiae BY4741 and L. plantarum HM23 for ethanol fermentation in repeated batch cultures. This mixed-specie biofilm was far abundantly formed and far resistant to washing compared with S. cerevisiae single biofilm. Adopting mixed-specie biofilm formed on cellulose beads as immobilized cells, we could produce enough ethanol from 10 or 20 % glucose during ten times repeated batch cultures for a duration of 10 days. Cell numbers of S. cerevisiae and L. plantarum during this period were stable. In mixed-specie biofilm system, though ethanol production was slightly lower compared to S. cerevisiae single-culture system due to by-production of lactate, pH was stably maintained under pH 4 without artificial control suggesting high resistance to contamination. Inoculated model contaminants, Escherichia coli and Bacillus subtilis, were excluded from the system in a short time. From the above results, it was indicated that the mixed-specie biofilm of S. cerevisiae and L. plantarum was a promising immobilized cell for ethanol fermentation for its ethanol productivity and robustness due to high resistance to contamination. PMID:23817789
Abe, Atsumu; Furukawa, Soichi; Watanabe, Shinya; Morinaga, Yasushi
Biofilm Formation by Pneumocystis spp.? �
Pneumocystis spp. can cause a lethal pneumonia in hosts with debilitated immune systems. The manner in which these fungal infections spread throughout the lung, the life cycles of the organisms, and their strategies used for survival within the mammalian host are largely unknown, due in part to the lack of a continuous cultivation method. Biofilm formation is one strategy used by microbes for protection against environmental assaults, for communication and differentiation, and as foci for dissemination. We posited that the attachment and growth of Pneumocystis within the lung alveoli is akin to biofilm formation. An in vitro system comprised of insert wells suspended in multiwell plates containing supplemented RPMI 1640 medium supported biofilm formation by P. carinii (from rat) and P. murina (from mouse).Dramatic morphological changes accompanied the transition to a biofilm. Cyst and trophic forms became highly refractile and produced branching formations that anastomosed into large macroscopic clusters that spread across the insert. Confocal microscopy revealed stacking of viable organisms enmeshed in concanavalin A-staining extracellular matrix. Biofilms matured over a 3-week time period and could be passaged. These passaged organisms were able to cause infection in immunosuppressed rodents. Biofilm formation was inhibited by farnesol, a quorum-sensing molecule in Candida spp., suggesting that a similar communication system may be operational in the Pneumocystis biofilms. Intense staining with a monoclonal antibody to the major surface glycoproteins and an increase in (1,3)-?-d-glucan content suggest that these components contributed to the refractile properties. Identification of this biofilm process provides a tractable in vitro system that should fundamentally advance the study of Pneumocystis.
Cushion, Melanie T.; Collins, Margaret S.; Linke, Michael J.
Biofilm formation in water cooling systems.
Biofilm formation on stainless steel samples immersed in cooling water has been evaluated by exposing metal samples to cooling seawater for 30 days. Anaerobic bacteria were then at 1.6 � 10(6)/cm(2), with sulphate-reducing species predominating. Aerobic bacteria and fungi were 2600 and 140/cm(2), respectively. After 60 days, numbers of aerobic microorganisms remained constant whereas the count of anaerobic microorganisms had increased to 1.8�10(9)/cm(2). Scanning electron microscopy showed the presence of morphologically different microorganisms in deposits and as a mucilaginous net. No signs of corrosion were detected on the stainless steel surface. PMID:24415316
Lutterbach, M T; de Fran�a, F P
Regulation of biofilm formation in Pseudomonas and Burkholderia species.
In the present review, we describe and compare the molecular mechanisms that are involved in the regulation of biofilm formation by Pseudomonas putida,?Pseudomonas fluorescens,?Pseudomonas aeruginosa and Burkholderia cenocepacia. Our current knowledge suggests that biofilm formation is regulated by cyclic diguanosine-5'-monophosphate (c-di-GMP), small RNAs (sRNA) and quorum sensing (QS) in all these bacterial species. The systems that employ c-di-GMP as a second messenger regulate the production of exopolysaccharides and surface proteins which function as extracellular matrix components in the biofilms formed by the bacteria. The systems that make use of sRNAs appear to regulate the production of exopolysaccharide biofilm matrix material in all these species. In the pseudomonads, QS regulates the production of extracellular DNA, lectins and biosurfactants which all play a role in biofilm formation. In B.?cenocepacia?QS regulates the expression of a large surface protein, lectins and extracellular DNA that all function as biofilm matrix components. Although the three regulatory systems all regulate the production of factors used for biofilm formation, the molecular mechanisms involved in transducing the signals into expression of the biofilm matrix components differ between the species. Under the conditions tested, exopolysaccharides appears to be the most important biofilm matrix components for P.?aeruginosa, whereas large surface proteins appear to be the most important biofilm matrix components for P.?putida,?P.?fluorescens, and B.?cenocepacia. PMID:24592823
Fazli, Mustafa; Almblad, Henrik; Rybtke, Morten Levin; Givskov, Michael; Eberl, Leo; Tolker-Nielsen, Tim
Biofilm formation of Bdellovibrio bacteriovorus host-independent derivatives.
Bdellovibrios are Gram-negative predatory bacteria which are ubiquitous to many environmental niches, including natural biofilms. In this study, host-independent (HI) variants of Bdellovibrio bacteriovorus 109J were isolated. Predation assays and genetic analysis confirmed that the selected HI variants are derivatives of B. bacteriovorus. When grown in microtiter plates, HI variants were able to form tenacious biofilms on the surface of the wells. HI biofilm formation on different surfaces, media and temperatures was examined. HI biofilm development was seen on all of the examined surfaces, with the most robust biofilm developing at 22 degrees C and in media supplemented with yeast extract. Biofilm detachment experiments revealed that the HI cells are firmly attached to the surface of the wells and are not easily removed by physical and chemical treatments. Treating the biofilm with proteinase K and DNase-I caused rapid detachment of the biofilm as well as inhibition of biofilm formation, suggesting that DNA and proteins are major components of the HI biofilm extracellular matrix. Our data suggest that under conditions that might favor the development of HI variants, such as a rich nutrient environment, Bdellovibrio facultative prey cells are capable of attaching to abiotic surfaces and forming biofilms. PMID:19223013
Medina, Adrian A; Kadouri, Daniel E
Adenoid Reservoir for Pathogenic Biofilm Bacteria?
Biofilms of pathogenic bacteria are present on the middle ear mucosa of children with chronic otitis media (COM) and may contribute to the persistence of pathogens and the recalcitrance of COM to antibiotic treatment. Controlled studies indicate that adenoidectomy is effective in the treatment of COM, suggesting that the adenoids may act as a reservoir for COM pathogens. To investigate the bacterial community in the adenoid, samples were obtained from 35 children undergoing adenoidectomy for chronic OM or obstructive sleep apnea. We used a novel, culture-independent molecular diagnostic methodology, followed by confocal microscopy, to investigate the in situ distribution and organization of pathogens in the adenoids to determine whether pathogenic bacteria exhibited criteria characteristic of biofilms. The Ibis T5000 Universal Biosensor System was used to interrogate the extent of the microbial diversity within adenoid biopsy specimens. Using a suite of 16 broad-range bacterial primers, we demonstrated that adenoids from both diagnostic groups were colonized with polymicrobial biofilms. Haemophilus influenzae was present in more adenoids from the COM group (P = 0.005), but there was no significant difference between the two patient groups for Streptococcus pneumoniae or Staphylococcus aureus. Fluorescence in situ hybridization, lectin binding, and the use of antibodies specific for host epithelial cells demonstrated that pathogens were aggregated, surrounded by a carbohydrate matrix, and localized on and within the epithelial cell surface, which is consistent with criteria for bacterial biofilms.
Nistico, L.; Kreft, R.; Gieseke, A.; Coticchia, J. M.; Burrows, A.; Khampang, P.; Liu, Y.; Kerschner, J. E.; Post, J. C.; Lonergan, S.; Sampath, R.; Hu, F. Z.; Ehrlich, G. D.; Stoodley, P.; Hall-Stoodley, L.
Boundaries for Biofilm Formation: Humidity and Temperature
Environmental conditions which define boundaries for biofilm production could provide useful ecological information for biofilm models. A practical use of defined conditions could be applied to the high-level nuclear waste repository at Yucca Mountain. Data for temperature and humidity conditions indicate that decreases in relative humidity or increased temperature severely affect biofilm formation on three candidate canister metals. Biofilms create
T. A. Else; C. R. Pantle; P. S. Amy
Biofilm formation in the industry: A review
Biofilm and biofouling refer to biological deposits on any surface. Biofilms consist of both microbes and their extracellular products, usually polysaccharides. The purpose of biofilm is to protect the microbes from hostile environments or to act as a trap for nutrient acquisition. Biofilm formation causes problems in many branches of industry, such as in industrial water systems and the medical
Gun Wirtanen
Staphylococcus aureus biofilm formation at the physiologic glucose concentration depends on the S. aureus lineage
BACKGROUND: Since bacteria embedded in biofilms are far more difficult to eradicate than planktonic infections, it would be useful to know whether certain Staphylococcus aureus lineages are especially involved in strong biofilm formation. For this reason, in vitro biofilm formation of 228 clinical S. aureus isolates of distinct clonal lineages was investigated. RESULTS: At 0.1% glucose, more than 60% of
Sander Croes; Ruud H Deurenberg; Marie-Louise L Boumans; Patrick S Beisser; Cees Neef; Ellen E Stobberingh
Observations on biofilm formation in industrial air-cooling units
Observations on biofilm formation in industrial air-cooling units were made over a 60-day operational period. Methods employed included: epifluorescent direct counts of water and slime samples, enumeration of culturable bacteria in water and slime samples, and ultrastructural observations of microbial attachment to formvar coated grids and epoxy resin blocks. Acridine orange direct counts and culturable counts of bacteria in water
C. A. Liebert; M. A. Hood; P. A. Winter; F. L. Singleton
Effect of alkaline pH on staphylococcal biofilm formation.
Biofilms are a serious problem, cause of severe inconvenience in the biomedical, food and industrial environment. Staphylococcus aureus and S. epidermidis are important pathogenic bacteria able to form thick and resistant biofilms on various surfaces. Therefore, strategies aimed at preventing or at least interfering with the initial adhesion and subsequent biofilm formation are a considerable achievement. The aim of this study was to evaluate the effect of alkaline pH on bacterial adhesion and further biofilm formation of S. aureus and S. epidermidis strains by biofilm biomass, cell-surface hydrophobicity, scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) analysis. The results demonstrated that the amount of biofilm biomass formed and the surface hydrophobicity were significantly less than what were observed at higher levels of pH. SEM and CLSM images revealed a poorly structured and very thin biofilm (2.5-3 times thinner than that of the controls). The inhibiting effect of the alkaline pH on the bacterial attachment impaired the normal development of biofilm that arrested at the microcolony stage. Alkaline formulations could be promising towards the control of bacterial colonization and therefore the reduction of the biofilm-related hazard. In the clinical setting, alkaline solutions or cleaners could be promising to prevent the bacterial colonization, by treating surfaces such as catheters or indwelling medical devices, reducing the risk of biofilm related infections. PMID:22882263
Nostro, Antonia; Cellini, Luigina; Di Giulio, Mara; D'Arrigo, Manuela; Marino, Andreana; Blanco, Anna Rita; Favaloro, Angelo; Cutroneo, Giuseppina; Bisignano, Giuseppe
Electron microscopic examination of wastewater biofilm formation and structural components.
This research documents in situ wastewater biofilm formation, structure, and physiochemical properties as revealed by scanning and transmission electron microscopy. Cationized ferritin was used to label anionic sites of the biofilm glycocalyx for viewing in thin section. Wastewater biofilm formation paralleled the processes involved in marine biofilm formation. Scanning electron microscopy revealed a dramatic increase in cell colonization and growth over a 144-h period. Constituents included a variety of actively dividing morphological types. Many of the colonizing bacteria were flagellated. Filaments were seen after primary colonization of the surface. Transmission electron microscopy revealed a dominant gram-negative cell wall structure in the biofilm constituents. At least three types of glycocalyces were observed. The predominant glycocalyx possessed interstices and was densely labeled with cationized ferritin. Two of the glycocalyces appeared to mediate biofilm adhesion to the substratum. The results suggest that the predominant glycocalyx of this thin wastewater biofilm serves, in part, to: (i) enclose the bacteria in a matrix and anchor the biofilm to the substratum and (ii) provide an extensive surface area with polyanionic properties. Images
Eighmy, T T; Maratea, D; Bishop, P L
Effects of patterned topography on biofilm formation
NASA Astrophysics Data System (ADS)
Bacterial biofilms are a population of bacteria attached to each other and irreversibly to a surface, enclosed in a matrix of self-secreted polymers, among others polysaccharides, proteins, DNA. Biofilms cause persisting infections associated with implanted medical devices and hospital acquired (nosocomial) infections. Catheter-associated urinary tract infections (CAUTIs) are the most common type of nosocomial infections accounting for up to 40% of all hospital acquired infections. Several different strategies, including use of antibacterial agents and genetic cues, quorum sensing, have been adopted for inhibiting biofilm formation relevant to CAUTI surfaces. Each of these methods pertains to certain types of bacteria, processes and has shortcomings. Based on eukaryotic cell topography interaction studies and Ulva linza spore studies, topographical surfaces were suggested as a benign control method for biofilm formation. However, topographies tested so far have not included a systematic variation of size across basic topography shapes. In this study patterned topography was systematically varied in size and shape according to two approaches 1) confinement and 2) wetting. For the confinement approach, using scanning electron microscopy and confocal microscopy, orienting effects of tested topography based on staphylococcus aureus (s. aureus) (SH1000) and enterobacter cloacae (e. cloacae) (ATCC 700258) bacterial models were identified on features of up to 10 times the size of the bacterium. Psuedomonas aeruginosa (p. aeruginosa) (PAO1) did not show any orientational effects, under the test conditions. Another important factor in medical biofilms is the identification and quantification of phenotypic state which has not been discussed in the literature concerning bacteria topography characterizations. This was done based on antibiotic susceptibility evaluation and also based on gene expression analysis. Although orientational effects occur, phenotypically no difference was observed between the patterned topography tested. Another potential strategy for biofilm control through patterned topography is based on the design of robust non-wetting surfaces with undercut feature geometries, characterized by 1) breakthrough pressure and 2) triple phase contact line model. It was found that height and presence of undercut had statistically significant effects, directly proportional to breakthrough pressures, whereas extent of undercut did not. A predictive triple phase contact line model was also developed. (Full text of this dissertation may be available via the University of Florida Libraries web site. Please check http://www.uflib.ufl.edu/etd.html)
Vasudevan, Ravikumar
Characterization of biofilm formation on a humic material.
Biofilms are major sites of carbon cycling in streams. Therefore, it is crucial to improve knowledge about biofilms' structure and microbial composition to understand their contribution in the self-purification of surface water. The present work intends to study biofilm formation in the presence of humic substances (HSs) as a carbon source. Two biofilm flowcells were operated in parallel; one with synthetic stream water, displaying a background carbon concentration of 1.26+/-0.84 mg L(-1), the other with added HSs and an overall carbon concentration of 9.68+/-1.00 mg L(-1). From the biofilms' results of culturable and total countable cells, it can be concluded that the presence of HSs did not significantly enhance the biofilm cell density. However, the biofilm formed in the presence of HSs presented slightly higher values of volatile suspended solids (VSS) and protein. One possible explanation for this result is that HSs adsorbed to the polymeric matrix of the biofilm and were included in the quantification of VSS and protein. The microbial composition of the biofilm with addition of HSs was characterized by the presence of bacteria belonging to beta-Proteobacteria, Cupriavidus metallidurans and several species of the genus Ralstonia were identified, and gamma-Proteobacteria, represented by Escherichia coli. In the biofilm formed without HSs addition beta-Proteobacteria, represented by the species Variovorax paradoxus, and bacteria belonging to the group Bacteroidetes were detected. In conclusion, the presence of HSs did not significantly enhance biofilm cell density but influenced the bacterial diversity in the biofilm. PMID:18712549
Rodrigues, A L; Brito, A G; Janknecht, P; Silva, J; Machado, A V; Nogueira, R
Tight Modulation of Escherichia coli Bacterial Biofilm Formation through Controlled Expression of Adhesion Factors
Despite the economic and sanitary problems caused by harmful biofilms, biofilms are nonetheless used empirically in industrial environmental and bioremediation processes and may be of potential use in medical settings for interfering with pathogen development. Escherichia coli is one of the bacteria with which biofilm formation has been studied in great detail, and it is especially appreciated for biotechnology applications
Sandra Da Re; Benjamin Le Quere; Jean-Marc Ghigo; Christophe Beloin
Printed paper-based arrays as substrates for biofilm formation
The suitability of paper-based arrays for biofilm formation studies by Staphylococcus aureus is demonstrated. Laboratory-coated papers with different physicochemical properties were used as substrates. The array platform was fabricated by patterning the coated papers with vinyl-substituted polydimethylsiloxane (PDMS) -based ink. The affinity of bacteria onto the flexographically printed hydrophobic and smooth PDMS film was very low whereas bacterial adhesion and biofilm formation occurred preferentially on the unprinted areas, i.e. in the reaction arrays. The concentration of the attached bacteria was quantified by determining the viable colony forming unit (CFU/cm2) numbers. The distribution and the extent of surface coverage of the biofilms were determined by atomic force microscopy. In static conditions, the highest bacterial concentration and most highly organized biofilms were observed on substrates with high polarity. On a rough paper surface with low polarity, the biofilm formation was most hindered. Biofilms were effectively removed from a polar substrate upon exposure to (+)-dehydroabietic acid, an anti-biofilm compound.
Printed paper-based arrays as substrates for biofilm formation.
The suitability of paper-based arrays for biofilm formation studies by Staphylococcus aureus is demonstrated. Laboratory-coated papers with different physicochemical properties were used as substrates. The array platform was fabricated by patterning the coated papers with vinyl-substituted polydimethylsiloxane (PDMS) -based ink. The affinity of bacteria onto the flexographically printed hydrophobic and smooth PDMS film was very low whereas bacterial adhesion and biofilm formation occurred preferentially on the unprinted areas, i.e. in the reaction arrays. The concentration of the attached bacteria was quantified by determining the viable colony forming unit (CFU/cm(2)) numbers. The distribution and the extent of surface coverage of the biofilms were determined by atomic force microscopy. In static conditions, the highest bacterial concentration and most highly organized biofilms were observed on substrates with high polarity. On a rough paper surface with low polarity, the biofilm formation was most hindered. Biofilms were effectively removed from a polar substrate upon exposure to (+)-dehydroabietic acid, an anti-biofilm compound. PMID:25006538
M��tt�nen, Anni; Fallarero, Adyary; Kujala, Janni; Ihalainen, Petri; Vuorela, Pia; Peltonen, Jouko
Shewanella putrefaciens adhesion and biofilm formation on food processing surfaces.
Laboratory model systems were developed for studying Shewanella putrefaciens adhesion and biofilm formation under batch and flow conditions. S. putrefaciens plays a major role in food spoilage and may cause microbially induced corrosion on steel surfaces. S. putrefaciens bacteria suspended in buffer adhered readily to stainless steel surfaces. Maximum numbers of adherent bacteria per square centimeter were reached in 8 h at 25 degrees C and reflected the cell density in suspension. Numbers of adhering bacteria from a suspension containing 10(8) CFU/ml were much lower in a laminar flow system (modified Robbins device) (reaching 10(2) CFU/cm(2)) than in a batch system (reaching 10(7) CFU/cm(2)), and maximum numbers were reached after 24 h. When nutrients were supplied, S. putrefaciens grew in biofilms with layers of bacteria. The rate of biofilm formation and the thickness of the film were not dependent on the availability of carbohydrate (lactate or glucose) or on iron starvation. The number of S. putrefaciens bacteria on the surface was partly influenced by the presence of other bacteria (Pseudomonas fluorescens) which reduced the numbers of S. putrefaciens bacteria in the biofilm. Numbers of bacteria on the surface must be quantified to evaluate the influence of environmental factors on adhesion and biofilm formation. We used a combination of fluorescence microscopy (4',6'-diamidino-2-phenylindole staining and in situ hybridization, for mixed-culture studies), ultrasonic removal of bacteria from surfaces, and indirect conductometry and found this combination sufficient to quantify bacteria on surfaces. PMID:11319118
Bagge, D; Hjelm, M; Johansen, C; Huber, I; Gram, L
Colonization of aerobic biofilms by sulfate?reducing bacteria
The ability of the SRB Desulfovibrio desulfuricans to colonize aerobic heterotrophic or nitrifying biofilms on stainless steel surfaces was investigated by fluorescence in situ hybridization (FISH) in combination with confocal laser scanning microscopy (CLSM), the polymerase chain reaction (PCR) directed to the dissimilatory sulfite reductase and by oxygen microelectrodes. Biofilms of heterotrophic bacteria and of nitrifying bacteria pregrown on steel
Mary E Power; Jan Roelof Van Der Meer; Hauke Harms; Oskar Wanner
Chemically Specific Cellular Imaging of Biofilm Formation
This document and the accompanying manuscripts summarize the technical accomplishments for our one-year LDRD-ER effort. Biofilm forming microbes have existed on this planet for billions of years and make up 60% of the biological mass on earth. Such microbes exhibit unique biochemical pathways during biofilm formation and play important roles in human health and the environment. Microbial biofilms have been
J L Herberg; C Schaldach; J Horn; E Gjersing; R Maxwell
Identification of a Novel Benzimidazole That Inhibits Bacterial Biofilm Formation in a Broad-Spectrum Manner?
Bacterial biofilm formation causes significant industrial economic loss and high morbidity and mortality in medical settings. Biofilms are defined as multicellular communities of bacteria encased in a matrix of protective extracellular polymers. Because biofilms have a high tolerance for treatment with antimicrobials, protect bacteria from immune defense, and resist clearance with standard sanitation protocols, it is critical to develop new approaches to prevent biofilm formation. Here, a novel benzimidazole molecule, named antibiofilm compound 1 (ABC-1), identified in a small-molecule screen, was found to prevent bacterial biofilm formation in multiple Gram-negative and Gram-positive bacterial pathogens, including Pseudomonas aeruginosa and Staphylococcus aureus, on a variety of different surface types. Importantly, ABC-1 itself does not inhibit the growth of bacteria, and it is effective at nanomolar concentrations. Also, coating a polystyrene surface with ABC-1 reduces biofilm formation. These data suggest ABC-1 is a new chemical scaffold for the development of antibiofilm compounds.
Sambanthamoorthy, Karthik; Gokhale, Ankush A.; Lao, Weiwei; Parashar, Vijay; Neiditch, Matthew B.; Semmelhack, Martin F.; Lee, Ilsoon; Waters, Christopher M.
Inhibitory Effects of Lactoferrin on Growth and Biofilm Formation of Porphyromonas gingivalis and Prevotella intermedia
Lactoferrin (LF) is an iron-binding antimicrobial protein present in saliva and gingival crevicular fluids, and it is possibly associated with host defense against oral pathogens, including periodontopathic bacteria. In the present study, we evaluated the in vitro effects of LF-related agents on the growth and biofilm formation of two periodontopathic bacteria, Porphyromonas gingivalis and Prevotella intermedia, which reside as biofilms
Hiroyuki Wakabayashi; Koji Yamauchi; Tetsuo Kobayashi; Tomoko Yaeshima; Keiji Iwatsuki; Hiromasa Yoshie
Cigarette Smoke Increases Staphylococcus aureus Biofilm Formation via Oxidative Stress
The strong epidemiological association between cigarette smoke (CS) exposure and respiratory tract infections is conventionally attributed to immunosuppressive and irritant effects of CS on human cells. Since pathogenic bacteria such as Staphylococcus aureus are members of the normal microbiota and reside in close proximity to human nasopharyngeal cells, we hypothesized that bioactive components of CS might affect these organisms and potentiate their virulence. Using Staphylococcus aureus as a model organism, we observed that the presence of CS increased both biofilm formation and host cell adherence. Analysis of putative molecular pathways revealed that CS exposure decreased expression of the quorum-sensing agr system, which is involved in biofilm dispersal, and increased transcription of biofilm inducers such as sarA and rbf. CS contains bioactive compounds, including free radicals and reactive oxygen species, and we observed transcriptional induction of bacterial oxidoreductases, including superoxide dismutase, following exposure. Moreover, pretreatment of CS with an antioxidant abrogated CS-mediated enhancement of biofilms. Exposure of bacteria to hydrogen peroxide alone increased biofilm formation. These observations are consistent with the hypothesis that CS induces staphylococcal biofilm formation in an oxidant-dependent manner. CS treatment induced transcription of fnbA (encoding fibronectin binding protein A), leading to increased binding of CS-treated staphylococci to immobilized fibronectin and increased adherence to human cells. These observations indicate that the bioactive effects of CS may extend to the resident microbiota of the nasopharynx, with implications for the pathogenesis of respiratory infection in CS-exposed humans.
Kulkarni, Ritwij; Antala, Swati; Wang, Alice; Amaral, Fabio E.; Rampersaud, Ryan; LaRussa, Samuel J.; Planet, Paul J.
Bacterial survival and biofilm formation on conventional and antibacterial toothbrushes
The aim of this study was to investigate bacterial survival and biofilm formation on toothbrushes. Fifteen healthy volunteers each used a normal toothbrush and an antibacterial toothbrush of the same design for two separate 5 week periods. Bacteria were removed from the brush head by swabbing and mechanical agitation in 10 ml of tryptone soya broth, cultured aerobically on selective
R. L. Sammons; D. Kaur; P. Neal
Aminoglycoside inhibition of Staphylococcus aureus biofilm formation is nutrient dependent.
Biofilms represent microbial communities, encased in a self-produced matrix or extracellular polymeric substance. Microbial biofilms are likely responsible for a large proportion of clinically significant infections and the multicellular nature of biofilm existence has been repeatedly associated with antibiotic resistance. Classical in vitro antibiotic-susceptibility testing utilizes artificial growth media and planktonic microbes, but this method may not account for the variability inherent in environments subject to biofilm growth in vivo. Experiments were designed to test the hypothesis that nutrient concentration can modulate the antibiotic susceptibility of Staphylococcus aureus biofilms. Developing S. aureus biofilms initiated on surgical sutures, and in selected experiments planktonic cultures, were incubated for 16 h in 66?% tryptic soy broth, 0.2?% glucose (1� TSBg), supplemented with bactericidal concentrations of gentamicin, streptomycin, ampicillin or vancomycin. In parallel experiments, antibiotics were added to growth medium diluted one-third (1/3� TSBg) or concentrated threefold (3� TSBg). Following incubation, viable bacteria were enumerated from planktonic cultures or suture sonicates, and biofilm biomass was assayed using spectrophotometry. Interestingly, bactericidal concentrations of gentamicin (5 �g gentamicin ml(-1)) and streptomycin (32 �g streptomycin ml(-1)) inhibited biofilm formation in samples incubated in 1/3� or 1� TSBg, but not in samples incubated in 3� TSBg. The nutrient dependence of aminoglycoside susceptibility is not only associated with biofilm formation, as planktonic cultures incubated in 3� TSBg in the presence of gentamicin also showed antibiotic resistance. These findings appeared specific for aminoglycosides because biofilm formation was inhibited in all three growth media supplemented with bactericidal concentrations of the cell wall-active antibiotics, ampicillin and vancomycin. Additional experiments showed that the ability of 3� TSBg to overcome the antibacterial effects of gentamicin was associated with decreased uptake of gentamicin by S. aureus. Uptake is known to be decreased at low pH, and the kinetic change in pH of growth medium from biofilms incubated in 5 �g gentamicin ml(-1) in the presence of 3� TSBg was decreased when compared with pH determinations from biofilms formed in 1/3� or 1� TSBg. These studies underscore the importance of environmental factors, including nutrient concentration and pH, on the antibiotic susceptibility of S. aureus planktonic and biofilm bacteria. PMID:24696518
Henry-Stanley, Michelle J; Hess, Donavon J; Wells, Carol L
Imaging bacteria and biofilms on hardware and periprosthetic tissue in orthopedic infections.
Infection is a major complication of total joint arthroplasty (TJA) surgery, and even though it is now as low as 1 % in some hospitals, the increasing number of primary surgeries translates to tens of thousands of revisions due to prosthetic joint infection (PJI). In many cases the only solution is revision surgery in which the hardware is removed. This process is extremely long and painful for patients and is a considerable financial burden for the health-care system. A significant proportion of the difficulties in diagnosis and treatment of PJI are associated with biofilm formation where bacteria attach to the surface of the prosthesis and periprosthetic tissue and build a 3-D biofilm community encased in an extracellular polymeric slime (EPS) matrix. Bacteria in biofilms have a low metabolic rate which is thought to be a major contributor to their recalcitrance to antibiotic treatment. The diagnosis of biofilm infections is difficult due to the fact that bacteria in biofilms are not readily cultured with standard clinical microbiology techniques. To identify and visualize in situ biofilm bacteria in orthopedic samples, we have developed protocols for the collection of samples in the operating room, for molecular fluorescent staining with 16S rRNA fluorescence in situ hybridization (FISH), and for imaging of samples using confocal laser scanning microscopy (CLSM). Direct imaging is the only method which can definitively identify biofilms on implants and complements both culture and culture-independent diagnostic methods. PMID:24664829
Nistico, Laura; Hall-Stoodley, Luanne; Stoodley, Paul
Prevention of biofilm formation by polymer modification
Bacterial biofilm formation on synthetic polymers plays an important role in industry and in modern medicine, leading, for example, to difficult-to-treat infections caused by colonized foreign bodies. Prevention of biofilm formation is a necessary step in the successful prophylaxis of such infections. One approach is to inhibit bacterial adherence by polymer surface modification. We have investigated polymer modification by glow
B Jansen; W Kohnen
Phylogenetic Relationships and Coaggregation Ability of Freshwater Biofilm Bacteria
Coaggregation between bacteria occurs when two or more genetically distinct strains interact by specific cell-cell recogni- tion (12). The phenomenon was first recognized between dif- ferent oral plaque-forming bacteria, where both intergeneric and intrageneric coaggregation occurs (11). Coaggregation also occurs between bacteria isolated from a freshwater biofilm (3, 19), and it has been suggested that coaggregation may also mediate in
Alex H. Rickard; Stephen A. Leach; Laurence S. Hall; Clive M. Buswell; Nicola J. High; Pauline S. Handley
Characterization of biofilm formation by Riemerella anatipestifer.
Riemerella anatipestifer (RA) causes epizootics of infectious disease in poultry and results in serious economic losses, especially for the duck industry. The present study focuses on understanding the biofilm-producing ability of RA strains in attempt to explain the intriguing persistence of RA post-infection on duck farms. Four RA serotype reference strains and 39 field RA isolates were measured for the biofilm formation by crystal violet staining. Eighteen out of the 43 RA strains produced biofilms. Furthermore, RA isolate CH3 was treated with carbohydrates (sucrose; glucose), disodium EDTA (EDTA), antibiotics (ampicillin; chloramphenicol) or detergent (Triton X-100) to determine the effect of the treatments on biofilm formation. Biofilm formation by RA isolate CH3 was independent of sucrose but significantly inhibited by 5% glucose and 0.1 mmol/L EDTA. Biofilmed CH3 culture (CH3 grown with a biofilm) was 5-31 times more resistant to the treatments of ampicillin, chloramphenicol or Triton X-100 than planktonic CH3 culture on the basis of minimal inhibitory concentration and minimal bactericidal concentration. The development and architecture of the biofilm formed by CH3 were also assessed using confocal laser scanning microscopy, scanning electron microscopy and fluorescence microscopy. In addition, animal experiment was performed to determine the median lethal doses (LD(50)) of three RA isolates with different biofilm formation abilities. Despite the result that virulence is strain-dependent as a result of various factors other than biofilm-producing ability, the fact that biofilmed isolate is more resistant to antibiotic and detergent treatments than planktonic isolate suggest that biofilm formation by RA may contribute to the persistent infections on duck farms. PMID:20226600
Hu, Qinghai; Han, Xiangan; Zhou, Xiaojin; Ding, Siyu; Ding, Chan; Yu, Shengqing
Staphylococcus aureus glucose-induced biofilm accessory proteins, GbaAB, influence biofilm formation in a PIA-dependent manner.
The Gram-positive bacteria Staphylococcus aureus and Staphylococcus epidermidis are capable of attaching to a biomaterial surface and forming resistant biofilms. The identification of biomolecular and regulatory factors involved in staphylococcal adhesion and biofilm formation is needed to understand biofilm-associated infection in humans. Here, we have identified a new operon, gbaAB (glucose induced biofilm accessory gene), that affects biofilm formation in S. aureus NCTC8325. Real-time reverse transcription PCR (RT-PCR) and electrophoretic mobility shift assay showed that GbaA and GbaB are transcribed from the same transcript, and GbaA directly inhibits the transcription of the gbaAB operon through self-repression. Our results indicated that the gbaA mutant displayed enhanced biofilm formation compared with the wild type. However, the gbaB and the gbaAB double mutant displayed reduced biofilm formation, suggesting that the gbaAB operon is involved in biofilm formation and that gbaB might be the key gene in biofilm regulation. Phenotypic analysis suggested that the gbaAB operon mediated biofilm formation of S. aureus at the multicellular aggregation stage rather than during initial attachment. In addition, real-time RT-PCR analysis showed that icaA was upregulated in the gbaA mutant and downregulated in the gbaB and gbaAB mutants compared with the wild type. In addition, the gbaA and the gbaB mutants affected the induction of biofilm formation by glucose. Our results suggest that the gbaAB operon is involved in the regulation of the multicellular aggregation step of S. aureus biofilm formation in response to glucose and that this regulation may be mediated through the ica operon. PMID:24836943
You, Yibo; Xue, Ting; Cao, Linyan; Zhao, Liping; Sun, Haipeng; Sun, Baolin
Regulation of biofilm formation in Escherichia coli K12: Effect of mutations in the genes HNS, STRA, LON, and RPON
More than 99% of bacteria exist in natural ecosystems as specifically organized biofilms adhering to solid surfaces. Biofilms\\u000a have a typical architecture and are enclosed in exopolymeric matrix. Bacteria living in biofilms are extremely resistant to\\u000a antibacterial factors. In this work, we studied the role of some global regulators of gene expression during biofilm formation\\u000a by cells of Escherichia coli
A. S. Belik; N. N. Tarasova; I. A. Khmel�
Biofilm Formation Derived from Ambient Air and the Characteristics of Apparatus
NASA Astrophysics Data System (ADS)
Biofilm is a kind of thin film on solidified matters, being derived from bacteria. Generally, planktonic bacteria float in aqueous environments, soil or air, most of which can be regarded as oligotrophic environments. Since they have to survive by instinct, they seek for nutrients that would exist on materials surfaces as organic matters. Therefore, bacteria attach materials surfaces reversibly. The attachment and detachment repeat for a while and finally, they attach on them irreversibly and the number of bacteria on them increases. At a threshold number, bacteria produce polymeric matters at the same time by quorum sensing mechanism and the biofilm produces on material surfaces. The biofilm produced in that way generally contains water (more than 80%), EPS (Exopolymeric Substance) and bacteria themselves. And they might bring about many industrial problems, fouling, corrosion etc. Therefore, it is very important for us to control and prevent the biofilm formation properly. However, it is generally very hard to produce biofilm experimentally and constantly in ambient atmosphere on labo scale. The authors invented an apparatus where biofilm could form on specimen's surfaces from house germs in the ambient air. In this experiment, we investigated the basic characteristics of the apparatus, reproducibility, the change of biofilm with experimental time, the quality change of water for biofilm formation and their significance for biofilm research.
Kanematsu, H.; Kougo, H.; Kuroda, D.; Itho, H.; Ogino, Y.; Yamamoto, Y.
Effect of Biocides on Biofilm Bacteria from Dental Unit Water Lines
Microbial biofilm formation in dental unit water lines (DUWL) is a phenomenon that has been recognized for nearly four decades.\\u000a Water delivered by DUWL can harbor high numbers of bacteria, including opportunistic pathogens. Biofilms on tubing within\\u000a DUWL may serve as a reservoir for these microorganisms and should therefore be controlled. In this study, the effects of eight\\u000a biocides were
I. Liaqat; A. N. Sabri
Desiccation tolerance of iron bacteria biofilms on Mars regolith simulants
NASA Astrophysics Data System (ADS)
Iron oxidizing bacteria play an important role in the geological redox cycling of iron on earth. The redox change between Fe(II) and Fe(III) can be used for biological energy production [1]. Therefore iron oxidation in the iron rich martian soils may be or may have been microbially mediated. The microbial conversion of iron is considered to be an ancient form of metabolism [2], so it might have evolved on Mars as well. However, to exist in recent martian soils, bacteria must be able to endure dry and cold conditions. Neutrophilic iron oxidizers can be found in various iron rich aquatic environments, where they lead to the precipitation of insoluble ferric hydroxides. Some of these environments fall temporarily dry, what could have led to an adaptation to desiccation by bacteria, existing there. One strategy of iron bacteria to endure drought stress might be the formation of biofilms by excreting Extracellular Polymeric Substances (EPS). The deposition of iron hydroxides could enable them to endure dry conditions as well. For our experiments, neutrophilic iron oxidizing bacteria have been isolated from a creek in Bad Salzhausen/Hesse and temporarily drying out pools in Tierra del Fuego. Strains from aquatic environments in the national park "Unteres Odertal" and from water wells in Berlin/Brandenburg are included in the tests as well. In desiccation experiments, the capability of iron bacteria to tolerate dry conditions are investigated. The aim of our first experiment is the adaptation to dry conditions. Biofilms of 15 strains are grown on ceramic beads in liquid medium containing complexed Fe(II), established biofilms contain Fe(III) precipitates. The cultures are desiccated in a sterile airflow until the weight of the cultures remained constant. After a desiccation period of 9 h up to 7 d, the beads are transferred to fresh liquid medium. Adapted strains are used in further desiccation experiments, where biofilms are grown on two martian regolith simulants. These mineral mixtures were developed and produced by the Naturkundemuseum Berlin according to recent data of Mars research missions [3, 4, 5, 6, 7]. The minerals are attached to object slides with potassium silicate and biofilms are grown on the mineral surface. The biofilms are quantified by cell counting and the structure is evaluated by epifluorescence microscopy. After desiccation in a sterile airflow, the survival of cells is determined by fluorescence staining. Acknowledgements This research was supported by the Helmholtz Association through the research alliance "Planetary Evolution and Life". References [1] Weber, K. A. et al. (2006). Microorganisms pumping iron: anaerobic microbial iron oxidation and reduction. Nature Reviews Microbiology 4: 752-764. [2] Vargas, M. et al. (1998). Microbiological evidence for Fe(III) reduction on early Earth. Nature 395: 65-67. [3] Bibring, J.-P., Y. Langevin, et al. (2005). Mars surface diversity as revealed by the OMEGA/Mars express observations. Science 307(5715): 1576-1581. [4] Bibring, J.-P., S. W. Squyres, et al. (2006). Merging Views on Mars. Science 313(5795): 1899-1901. [5] Chevrier, V. and P. E. Math� (2007). Mineralogy and evolution of the surface of Mars: A review. Planetary and Space Science 55(3): 289-314. [6] McCollom, T. M. and B. M. Hynek (2005). A volcanic environment for bedrock diagenesis at Meridiani Planum on Mars. Nature 438(7071): 1129-1131. [7] Poulet, F., J. P. Bibring, et al. (2005). Phyllosilicates on Mars and implications for early martian climate. Nature 438(7068): 623-627.
Feyh, Nina; Szewzyk, Ulrich
ANTIFOULING ACTIVITY OF BACTERIA ASSOCIATED WITH SOFT CORAL Sarcophyton sp. AGAINST MARINE BIOFILM FORMING BACTERIA
Marine bacteria associated with soft coral Sarcophy ton sp collected from vicinity of Peucang island, Ujung Kulon, West Java, were successfully s creened for antifouling activity against marine biofilm-forming bacteria isolated from the surround ing colonies of Sarcophyton sp. Six bacterial isolates were found to inhibit the growth of at lea st one of 7 biofilm-forming isolates. The most act
Agus Sabdono; Ocky Karna Radjasa
Biofilm formation of Malassezia pachydermatis from dogs.
Yeasts of the genus Malassezia are commensals of the normal skin microbial flora of humans and animals. These yeasts may become pathogenic under certain circumstances and their pathogenic role may be related to host immune system as well to yeast virulence factors (e.g., phospholipase production and biofilm formation). This study aims to evaluate the in vitro ability of M. pachydermatis strains to produce biofilm, and its relationship with phospholipase activity and the genetic make-up of isolates from lesioned (n=32) and healthy (n=30) dog skin. The production of biofilm was determined by crystal violet staining and scanning electronic microscopy (SEM). Biofilm was produced by almost all M. pachydermatis isolates (95.2%) from dogs with and without skin lesions at variable level and different structure. At the SEM, biofilm matrix presented adhering blastoconidia clustered in multi- or monolayer structures with variable quantity of extracellular production. Of the three genotypes detected, genotype B showed the lowest ability to produce biofilm. Of the 59 isolates producing biofilm, 33 (55.9%) produced phospholipase, with a higher biofilm formation (p<0.05) in strains collected from animals with skin lesions. It is here suggested that phospholipase production might act in synergism with the biofilm formation by inducing or exacerbating skin lesions in dogs. The results provide evidences for a better understanding of the interactions between yeasts and host immune system, toward revealing the pathogenicity of M. pachydermatis in animals. PMID:22682201
Figueredo, Luciana A; Cafarchia, Claudia; Desantis, Salvatore; Otranto, Domenico
The interconnection between biofilm formation and horizontal gene transfer.
Recent research has revealed that horizontal gene transfer and biofilm formation are connected processes. Although published research investigating this interconnectedness is still limited, we will review this subject in order to highlight the potential of these observations because of their believed importance in the understanding of the adaptation and subsequent evolution of social traits in bacteria. Here, we discuss current evidence for such interconnectedness centred on plasmids. Horizontal transfer rates are typically higher in biofilm communities compared with those in planktonic states. Biofilms, furthermore, promote plasmid stability and may enhance the host range of mobile genetic elements that are transferred horizontally. Plasmids, on the other hand, are very well suited to promote the evolution of social traits such as biofilm formation. This, essentially, transpires because plasmids are independent replicons that enhance their own success by promoting inter-bacterial interactions. They typically also carry genes that heighten their hosts' direct fitness. Furthermore, current research shows that the so-called mafia traits encoded on mobile genetic elements can enforce bacteria to maintain stable social interactions. It also indicates that horizontal gene transfer ultimately enhances the relatedness of bacteria carrying the mobile genetic elements of the same origin. The perspective of this review extends to an overall interconnectedness between horizontal gene transfer, mobile genetic elements and social evolution of bacteria. PMID:22444301
Madsen, Jonas Stenl�kke; Burm�lle, Mette; Hansen, Lars Hestbjerg; S�rensen, S�ren Johannes
Biofilm Formation in Microscopic Double Emulsion Droplets
NASA Astrophysics Data System (ADS)
In natural, medical, and industrial settings, there exist surface-associated communities of bacteria known as biofilms. These highly structured films are composed of bacterial cells embedded within self-produced extracellular matrix, usually composed of exopolysaccharides, proteins, and nucleic acids; this matrix serves to protect the bacterial community from antibiotics and environmental stressors. Here, we form biofilms encapsulated within monodisperse, microscopically-sized double emulsion droplets using microfluidics. The bacteria self-organize at the inner liquid-liquid droplet interfaces, multiply, and differentiate into extracellular matrix-producing cells, forming manifold three-dimensional shell-within-a-shell structures of biofilms, templated upon the inner core of spherical liquid droplets. By using microfluidics to encapsulate bacterial cells, we have the ability to view individual cells multiplying in microscopically-sized droplets, which allows for high-throughput analysis in studying the genetic program leading to biofilm development, or cell signaling that induces differentiation.
Chang, Connie; Weitz, David
The contribution of cell-cell signaling and motility to bacterial biofilm formation
Many bacteria grow attached to a surface as biofilms. Several factors dictate biofilm formation, including responses by the colonizing bacteria to their environment. Here we review how bacteria use cell-cell signaling (also called quorum sensing) and motility during biofilm formation. Specifically, we describe quorum sensing and surface motility exhibited by the bacterium Pseudomonas aeruginosa, a ubiquitous environmental organism that acts as an opportunistic human pathogen in immunocompromised individuals. P. aeruginosa uses acyl-homoserine lactone signals during quorum sensing to synchronize gene expression important to the production of polysaccharides, rhamnolipid, and other virulence factors. Surface motility affects the assembly and architecture of biofilms, and some aspects of motility are also influenced by quorum sensing. While some genes and their function are specific to P. aeruginosa, many aspects of biofilm development can be used as a model system to understand how bacteria differentially colonize surfaces.
Shrout, Joshua D.; Tolker-Nielsen, Tim; Givskov, Michael; Parsek, Matthew R.
Identification of biofilm formation by Mycoplasma gallisepticum.
Mycoplasma gallisepticum is the causative agent of chronic respiratory disease in chickens and of infectious sinusitis in turkeys, chickens, game birds, pigeons, and passerine birds of all ages. This study investigated the biofilm-producing ability of M. gallisepticum strains in an attempt to explain its intriguing persistence in commercial flocks. Eleven strains of M. gallisepticum were investigated for their biofilm formation, which varied considerably. Strains Nobilis MG 6/85, S(6) (P(5) and P(20)), D(9604), and SU(15) were strong biofilm producers. Strains R(low) (P(10) and P(100)), NCL, CG(5), YL(4), and F were weak biofilm producers. Strains Vaxsafe MG ts-11 and F(36) did not produce biofilm as verified using a crystal violet staining assay. In addition, highly differentiated biofilm structures of strain Nobilis MG 6/85 with characteristic stacks and channels were observed under confocal scanning laser microscopy and scanning electron microscopy. The carbohydrates (sucrose, glucose), disodium ethylenediaminetetraacetic acid (EDTA), antibiotics (tetracycline, gentamicin), or detergent (Triton X-100) were further used to determine their effects on biofilm formation. Biofilm formation was significantly inhibited by 5% sucrose and 5 mmol/L EDTA. Compared with the planktonic mycoplasma, these biofilm-grown cultures were more resistant to tetracycline, gentamicin, and Triton X-100 treatments. Furthermore, real-time reverse transcriptase-polymerase chain reaction was performed to investigate the transcription of several genes that may be associated with biofilm formation. The results indicated that the transcriptions of some genes in the biofilm-grown cells were markedly decreased, including vlhA3.03, csmC, hatA, gapA, neuraminidase, and mgc2. Our results will benefit further research on the persistence of M. gallisepticum infections. PMID:22840542
Chen, Hongjun; Yu, Shengqing; Hu, Meirong; Han, Xiangan; Chen, Danqing; Qiu, Xusheng; Ding, Chan
Molecular Identification and Biofilm-Forming Ability of Culturable Aquatic Bacteria In Microbial Biofilms Formed in Drinking Water Distribution Networks
Drinking water distribution networks are known to harbor microbial biofilms. The aim of the present work is to (i) identify the culturable bacteria presented in the drinking-water distribution network, (ii) investigate the ability of isolated bacteria to form biofilm under some environmental stress conditions and some eliminating or removing treatments. To achieve it, 57 strains were isolated from biofilm (43
Hesham Elhariry; Youssuf Gherbawy; Bahig El-Deeb; Abdullah Altalhi
A Stochastic Mechanism for Biofilm Formation by Mycoplasma pulmonis
Bacterial biofilms are communities of bacteria that are enclosed in an extracellular matrix. Within a biofilm the bacteria are protected from antimicrobials, environmental stresses, and immune responses from the host. Biofilms are often believed to have a highly developed organization that is derived from differential regulation of the genes that direct the synthesis of the extracellular matrix and the attachment
Warren L. Simmons; Jeffrey R. Bolland; James M. Daubenspeck; Kevin Dybvig
Adhesion and biofilm formation of Mycoplasma pneumoniae on an abiotic surface.
We demonstrated that when M. pneumoniae was grown on an abiotic surface of either glass or polystyrene with a serum-containing medium, the bacteria adhered to the surface and formed highly differentiated volcano-like biofilm structures. As adherence to the surface and/or biofilm formation was totally inhibited by anti-P1 polyclonal monospecific antibodies, we suggest that the adherence of M. pneumoniae to the abiotic surface and/or biofilm formation is associated with P1, the major tip organelle protein of this organism. Furthermore, adherence and/or biofilm formation was markedly inhibited by treating the serum component of the growth medium with neuraminidase or by growing the bacteria in the presence of sialyllactose, suggesting that the initial step in the adherence to and/or biofilm formation by M. pneumoniae on an abiotic surface is the interaction of the bacterium through its tip organelle with sialic acid residues of serum glycoproteins. PMID:21879294
Kornspan, Jonathan D; Tarshis, Mark; Rottem, Shlomo
Phylogenetic Relationships and Coaggregation Ability of Freshwater Biofilm Bacteria
Nineteen numerically dominant heterotrophic bacteria from a freshwater biofilm were identified by 16S ribosomal DNA gene sequencing, and their coaggregation partnerships were determined. Phylogenetic trees showed that both distantly related and closely related strains coaggregated at intergeneric, intrageneric, and intraspecies levels. One strain, Blastomonas natatoria 2.1, coaggregated with all 18 other strains and may function as a bridging organism in biofilm development.
Rickard, Alex H.; Leach, Stephen A.; Hall, Laurence S.; Buswell, Clive M.; High, Nicola J.; Handley, Pauline S.
Fourier transform-infrared spectroscopic methods for microbial ecology: analysis of bacteria, bacteria-polymer mixtures and biofilms
NASA Technical Reports Server (NTRS)
Fourier transform-infrared (FT-IR) spectroscopy has been used to rapidly and nondestructively analyze bacteria, bacteria-polymer mixtures, digester samples and microbial biofilms. Diffuse reflectance FT-IR (DRIFT) analysis of freeze-dried, powdered samples offered a means of obtaining structural information. The bacteria examined were divided into two groups. The first group was characterized by a dominant amide I band and the second group of organisms displayed an additional strong carbonyl stretch at approximately 1740 cm-1. The differences illustrated by the subtraction spectra obtained for microbes of the two groups suggest that FT-IR spectroscopy can be utilized to recognize differences in microbial community structure. Calculation of specific band ratios has enabled the composition of bacteria and extracellular or intracellular storage product polymer mixtures to be determined for bacteria-gum arabic (amide I/carbohydrate C-O approximately 1150 cm-1) and bacteria-poly-beta-hydroxybutyrate (amide I/carbonyl approximately 1740 cm-1). The key band ratios correlate with the compositions of the material and provide useful information for the application of FT-IR spectroscopy to environmental biofilm samples and for distinguishing bacteria grown under differing nutrient conditions. DRIFT spectra have been obtained for biofilms produced by Vibrio natriegens on stainless steel disks. Between 48 and 144 h, an increase in bands at approximately 1440 and 1090 cm-1 was seen in FT-IR spectra of the V. natriegens biofilm. DRIFT spectra of mixed culture effluents of anaerobic digesters show differences induced by shifts in input feedstocks. The use of flow-through attenuated total reflectance has permitted in situ real-time changes in biofilm formation to be monitored and provides a powerful tool for understanding the interactions within adherent microbial consortia.
Nichols, P. D.; Henson, J. M.; Guckert, J. B.; Nivens, D. E.; White, D. C.
Agrobacterium-Host Attachment and Biofilm Formation
Physical association with host plant tissue is a prerequisite to Agrobacterium tumefaciens infection and subsequent disease. Mechanisms of tissue adherence have been extensively studied in mammalian pathogens, but\\u000a less so in plant-associated bacteria. Cells of A. tumefaciens often attach to plant tissue by a single pole. In the appropriate environment, these attached bacteria eventually develop\\u000a into multicellular assemblies called biofilms,
Clay Fuqua
Exopolysaccharide Productivity and Biofilm Phenotype on Oral Commensal Bacteria as Pathogenesis of Chronic Periodontitis.
National Technical Information Service (NTIS)
Exopolysaccharide (EPS) productivities in many bacteria have been associated with pathogenicity in mammalian hosts as providing extracellular matrices to form biofilm (Costerton et al., 1995). Bacteria assuming biofilm- forming capacity have enormous adva...
C. Mashimo H. Maruyama K. Yamane T. Nambu T. Yamanaka
Probiotic Lactobacilli Interfere with Streptococcus mutans Biofilm Formation In Vitro
In clinical studies, probiotic bacteria have decreased the counts of salivary mutans streptococci (MS). We compared the effects\\u000a of probiotic Lactobacillus strains on the biofilm formation of Streptococcus mutans. The bacterial strains used included four S. mutans strains (reference strains NCTC 10449 and Ingbritt and clinical isolates 2366 and 195) and probiotic strains Lactobacillus rhamnosus GG, L. plantarum 299v, and
Eva M. SoderlingAino; Aino M. Marttinen; Anna L. Haukioja
Biofilm formation on stainless steels in Arabian Gulf water
Metals immersed in natural seawater are covered with a thin, slimy film of micro-organisms and their biological secretion commonly referred to as biofilm. Biofilms cause the ennobling of the free corrosion potentials and increase the susceptibility of pitting- and crevice corrosion. Biofilm formation from Arabian Gulf water has not been reported before. Biofilm formation on 7 different types of Mo-containing
A. M. Shams El Din; T. M. H. Saber; A. A. Hammoud
Curli synthesis and biofilm formation in enteric bacteria are controlled by a dynamic small RNA module made up of a pseudoknot assisted by an RNA chaperone
RydC pseudoknot aided by Hfq is a dynamic regulatory module. We report that RydC reduces expression of curli-specific gene D transcription factor required for adhesion and biofilm production in enterobacteria. During curli formation, csgD messenger RNA (mRNA) synthesis increases when endogenous levels of RydC are lacking. In Escherichia coli and Salmonella enterica, stimulation of RydC expression also reduces biofilm formation by impairing curli synthesis. Inducing RydC early on in growth lowers CsgA, -B and -D protein and mRNA levels. RydC�s 5?-domain interacts with csgD mRNA translation initiation signals to prevent initiation. Translation inhibition occurs by an antisense mechanism, blocking the translation initiation signals through pairing, and that mechanism is facilitated by Hfq. Although Hfq represses csgD mRNA translation without a small RNA (sRNA), it forms a ternary complex with RydC and facilitates pseudoknot unfolding to interact with the csgD mRNA translation initiation signals. RydC action implies Hfq-assisted unfolding and mRNA rearrangements, but once the pseudoknot is disrupted, Hfq is unnecessary for regulation. RydC is the sixth sRNA that negatively controls CsgD synthesis. Hfq induces structural changes in the mRNA domains targeted by these six sRNAs. What we describe is an ingenious process whereby pseudoknot opening is orchestrated by a chaperone to allow RNA control of gene expression.
Bordeau, Valerie; Felden, Brice
Curli synthesis and biofilm formation in enteric bacteria are controlled by a dynamic small RNA module made up of a pseudoknot assisted by an RNA chaperone.
RydC pseudoknot aided by Hfq is a dynamic regulatory module. We report that RydC reduces expression of curli-specific gene D transcription factor required for adhesion and biofilm production in enterobacteria. During curli formation, csgD messenger RNA (mRNA) synthesis increases when endogenous levels of RydC are lacking. In Escherichia coli and Salmonella enterica, stimulation of RydC expression also reduces biofilm formation by impairing curli synthesis. Inducing RydC early on in growth lowers CsgA, -B and -D protein and mRNA levels. RydC's 5'-domain interacts with csgD mRNA translation initiation signals to prevent initiation. Translation inhibition occurs by an antisense mechanism, blocking the translation initiation signals through pairing, and that mechanism is facilitated by Hfq. Although Hfq represses csgD mRNA translation without a small RNA (sRNA), it forms a ternary complex with RydC and facilitates pseudoknot unfolding to interact with the csgD mRNA translation initiation signals. RydC action implies Hfq-assisted unfolding and mRNA rearrangements, but once the pseudoknot is disrupted, Hfq is unnecessary for regulation. RydC is the sixth sRNA that negatively controls CsgD synthesis. Hfq induces structural changes in the mRNA domains targeted by these six sRNAs. What we describe is an ingenious process whereby pseudoknot opening is orchestrated by a chaperone to allow RNA control of gene expression. PMID:24489123
Bordeau, Val�rie; Felden, Brice
Biofilm Formation by Staphylococcus haemolyticus
Infections due to coagulase-negative staphylococci (CoNS) most frequently occur after the implantation of medical devices and are attributed to the biofilm-forming potential of CoNS. Staphylococcus haemolyticus is the second most frequently isolated CoNS from patients with hospital-acquired infections. There is only limited knowledge of the nature of S. haemolyticus biofilms. The aim of this study was to characterize S. haemolyticus
Elizabeth Gladys; Aarag Fredheim; Claus Klingenberg; Holger Rohde; Stephanie Frankenberger; Peter Gaustad; Trond Fl�gstad; Johanna Ericson Sollid
Fluorescence-based quasicontinuous and in situ monitoring of biofilm formation dynamics in natural marine environments.
Analyzing the dynamics of biofilm formation helps to deepen our understanding of surface colonization in natural environments. While methods for screening biofilm formation in the laboratory are well established, studies in marine environments have so far been based upon destructive analysis of individual samples and provide only discontinuous snapshots of biofilm establishment. In order to explore the development of biofilm over time and under various biotic and abiotic conditions, we applied a recently developed optical biofilm sensor to quasicontinuously analyze marine biofilm dynamics in situ. Using this technique in combination with microscope-assisted imaging, we investigated biofilm formation from its beginning to mature multispecies biofilms. In contrast to laboratory studies on biofilm formation, a smooth transition from initial attachment to colony formation and exponential growth could not be observed in the marine environment. Instead, initial attachment was followed by an adaptation phase of low growth and homogeneously distributed solitary bacterial cells. Moreover, we observed a diurnal variation of biofilm signal intensity, suggesting a transient state of biofilm formation of bacteria. Overall, the biofilm formation dynamics could be modeled by three consecutive development stages attributed to initial bacterial attachment, bacterial growth, and attachment and growth of unicellular eukaryotic microorganisms. Additional experiments showed that the presence of seaweed considerably shortened the adaptation phase in comparison with that on control surfaces but yielded similar growth rates. The outlined examples highlight the advantages of a quasicontinuous in situ detection that enabled, for the first time, the exploration of the initial attachment phase and the diurnal variation during biofilm formation in natural ecosystems. PMID:24727266
Fischer, M; Friedrichs, G; Lachnit, T
Murine solid tumours as a novel model to study bacterial biofilm formation in vivo.
Bacteria of many species are able to invade and colonize solid tumours in mice. We have focused on Salmonella enterica serovar Typhimurium. Detailed analysis revealed that such tumour-invading Salmonella form biofilms, thus providing a versatile in vivo test system for studying bacterial phenotypes and host-pathogen interactions. It appears that biofilm formation by S.�typhimurium is induced as a defence against the immune system of the host, and in particular against neutrophils. Further, we extended our work to the clinically more relevant biofilm infection by Pseudomonas aeruginosa. The induction of P.�aeruginosa biofilms in neoplastic tissue appears to be elicited as a reaction against the immune system. Reconstitution experiments reveal that T cells are responsible for biofilm induction. Isogenic mutants that are no longer able to form biofilms can be used for comparison studies to determine antimicrobial resistance, especially therapeutic efficacy against P.�aeruginosa located in biofilms. PMID:24724621
Pawar, V; Crull, K; Komor, U; Kasnitz, N; Frahm, M; Kocijancic, D; Westphal, K; Leschner, S; Wolf, K; Loessner, H; Rohde, M; H�ussler, S; Weiss, S
Bacterial aggregation and biofilm formation in a vortical flow.
Bacterial aggregation and patchiness play an important role in a variety of ecological processes such as competition, adaptation, epidemics, and succession. Here, we demonstrate that hydrodynamics of their environment can lead to their aggregation. This is specially important since microbial habitats are rarely at rest (e.g., ocean, blood stream, flow in porous media, and flow through membrane filtration processes). In order to study the dynamics of bacterial collection in a vortical flow, we utilize a microfluidic system to mimic some of the important microbial conditions at ecologically relevant spatiotemporal scales. We experimentally demonstrate the formation of "ring"-shaped bacterial collection patterns and subsequently the formation of biofilm streamers in a microfluidic system. Acoustic streaming of a microbubble is used to generate a vortical flow in a microchannel. Due to bacteria's finite-size, the microorganisms are directed to closed streamlines and trapped in the vortical flow. The collection of bacteria in the vortices occurs in a matter of seconds, and unexpectedly, triggers the formation of biofilm streamers within minutes. Swimming bacteria have a competitive advantage to respond to their environmental conditions. In order to investigate the role of bacterial motility on the rate of collection, two strains of Escherichia coli bacteria with different motilities are used. We show that the bacterial collection in a vortical flow is strongly pronounced for high motile bacteria. PMID:24339847
Yazdi, Shahrzad; Ardekani, Arezoo M
Colonization of freshwater biofilms by nitrifying bacteria from activated sludge.
Effluents from wastewater treatment plants (WWTPs) containing micro-organisms and residual nitrogen can stimulate nitrification in freshwater streams. We hypothesized that different ammonia-oxidizing (AOB) and nitrite-oxidizing (NOB) bacteria present in WWTP effluents differ in their potential to colonize biofilms in the receiving streams. In an experimental approach, we monitored biofilm colonization by nitrifiers in ammonium- or nitrite-fed microcosm flumes after inoculation with activated sludge. In a field study, we compared the nitrifier communities in a full-scale WWTP and in epilithic biofilms downstream of the WWTP outlet. Despite substantially different ammonia concentrations in the microcosms and the stream, the same nitrifiers were detected by fluorescence in situ hybridization in all biofilms. Of the diverse nitrifiers present in the WWTPs, only AOB of the Nitrosomonas oligotropha/ureae lineage and NOB of Nitrospira sublineage I colonized the natural biofilms. Analysis of the amoA gene encoding the alpha subunit of ammonia monooxygenase of AOB revealed seven identical amoA sequence types. Six of these affiliated with the N.�oligotropha/ureae lineage and were shared between the WWTP and the stream biofilms, but the other shared sequence type grouped with the N.�europaea/eutropha and N.�communis lineage. Measured nitrification activities were high in the microcosms and the stream. Our results show that nitrifiers from WWTPs can colonize freshwater biofilms and confirm that WWTP-affected streams are hot spots of nitrification. PMID:23461285
Mu�mann, Marc; Ribot, Miquel; von Schiller, Daniel; Merbt, Stephanie N; Augspurger, Clemens; Karwautz, Clemens; Winkel, Matthias; Battin, Tom J; Mart�, Eug�nia; Daims, Holger
Marine Biofilm Bacteria Evade Eukaryotic Predation by Targeted Chemical Defense
Many plants and animals are defended from predation or herbivory by inhibitory secondary metabolites, which in the marine environment are very common among sessile organisms. Among bacteria, where there is the greatest metabolic potential, little is known about chemical defenses against bacterivorous consumers. An emerging hypothesis is that sessile bacterial communities organized as biofilms serve as bacterial refuge from predation. By testing growth and survival of two common bacterivorous nanoflagellates, we find evidence that chemically mediated resistance against protozoan predators is common among biofilm populations in a diverse set of marine bacteria. Using bioassay-guided chemical and genetic analysis, we identified one of the most effective antiprotozoal compounds as violacein, an alkaloid that we demonstrate is produced predominately within biofilm cells. Nanomolar concentrations of violacein inhibit protozoan feeding by inducing a conserved eukaryotic cell death program. Such biofilm-specific chemical defenses could contribute to the successful persistence of biofilm bacteria in various environments and provide the ecological and evolutionary context for a number of eukaryote-targeting bacterial metabolites.
Matz, Carsten; Webb, Jeremy S.; Schupp, Peter J.; Phang, Shui Yen; Penesyan, Anahit; Egan, Suhelen; Steinberg, Peter; Kjelleberg, Staffan
2D Model for Diffusion of Oxygen with Biochemical Reaction During Biofilm Formation Process in Static Aqueous Medium
A 2D model that describes diffusion of oxygen with biochemical reaction during biofilm formation process in static aqueous medium is presented. The analysis is based on X60 steel placed at the bottom of a container containing produced water inoculated with Leptothrix discophora (iron-oxidizing bacteria). These bacteria form biofilms on the exposed surfaces of the metal. The biofilm-microorganisms absorb oxygen from
Y. T. Puyate; A. Rim-Rukeh
Implications of Biofilm Formation on Urological Devices
NASA Astrophysics Data System (ADS)
Despite millions of dollars and several decades of research targeted at their prevention and eradication, biofilm-associated infections remain the major cause of urological device failure. Numerous strategies have been aimed at improving device design, biomaterial composition, surface properties and drug delivery, but have been largely circumvented by microbes and their plethora of attachment, host evasion, antimicrobial resistance, and dissemination strategies. This is not entirely surprising since natural biofilm formation has been going on for millions of years and remains a major part of microorganism survival and evolution. Thus, the fact that biofilms develop on and in the biomaterials and tissues of humans is really an extension of this natural tendency and greatly explains why they are so difficult for us to combat. Firstly, biofilm structure and composition inherently provide a protective environment for microorganisms, shielding them from the shear stress of urine flow, immune cell attack and some antimicrobials. Secondly, many biofilm organisms enter a metabolically dormant state that renders them tolerant to those antibiotics and host factors able to penetrate the biofilm matrix. Lastly, the majority of organisms that cause biofilm-associated urinary tract infections originate from our own oral cavity, skin, gastrointestinal and urogenital tracts and therefore have already adapted to many of our host defenses. Ultimately, while biofilms continue to hold an advantage with respect to recurrent infections and biomaterial usage within the urinary tract, significant progress has been made in understanding these dynamic microbial communities and novel approaches offer promise for their prevention and eradication. These include novel device designs, antimicrobials, anti-adhesive coatings, biodegradable polymers and biofilm-disrupting compounds and therapies.
Cadieux, Peter A.; Wignall, Geoffrey R.; Carriveau, Rupp; Denstedt, John D.
Comparative Study of Biofilm Formation in Pseudomonas aeruginosa Isolates from Patients of Lower Respiratory Tract Infection
Background: This study assessed biofilm formations of P.aeruginosa which was isolated from patients with Lower Respiratory Tract Infections (LRTIs). Objective: This study was conducted to compare different methods of biofilm formations seen in P. aeruginosa which was obtained from LRTI patients. Materials and Methods: In this cross-sectional study, we investigated a total of 80 P. aeruginosa isolates obtained from LRTI patients by different methods. Tube method (TM), tissue culture plate (TCP) method and modified tissue culture plate (MTCP) method. They were subjected to biofilm detection methods. Results: The MTCP method produced a higher accuracy ratio than TCP method. In terms of sensitivity and specificity, the MTCP method was considered to be superior to TM. We observed a higher antibiotic resistance in biofilm producing bacteria than in non-biofilm producers. Conclusion: In our study, MTCP was found to be more sensitive and specific method for biofilm detection than TCP and TM.
Saxena, Shivani; Garg, Rajiv; Singh, Mastan
Comparative Study of Biofilm Formation in Pseudomonas aeruginosa Isolates from Patients of Lower Respiratory Tract Infection.
Background: This study assessed biofilm formations of P.aeruginosa which was isolated from patients with Lower Respiratory Tract Infections (LRTIs). Objective: This study was conducted to compare different methods of biofilm formations seen in P. aeruginosa which was obtained from LRTI patients. Materials and Methods: In this cross-sectional study, we investigated a total of 80 P. aeruginosa isolates obtained from LRTI patients by different methods. Tube method (TM), tissue culture plate (TCP) method and modified tissue culture plate (MTCP) method. They were subjected to biofilm detection methods. Results: The MTCP method produced a higher accuracy ratio than TCP method. In terms of sensitivity and specificity, the MTCP method was considered to be superior to TM. We observed a higher antibiotic resistance in biofilm producing bacteria than in non-biofilm producers. Conclusion: In our study, MTCP was found to be more sensitive and specific method for biofilm detection than TCP and TM. PMID:24995174
Saxena, Shivani; Banerjee, Gopa; Garg, Rajiv; Singh, Mastan
Inhibition of Staphylococcus epidermidis Biofilm Formation by Rabbit Polyclonal Antibodies against the SesC Protein?
Several well-studied proteins with defined roles in Staphylococcus epidermidis biofilm formation are LPXTG motif-containing proteins. Here, we investigate the possible use of the LPXTG motif-containing protein SesC (S. epidermidis surface protein C; accession no. NP_765787) as a target for antibodies to prevent biofilm formation. In vitro and in a in vivo rat model of catheter infection, gene and protein expression analysis showed that SesC is expressed more strongly in biofilm-associated cells than in planktonic cells and is expressed particularly during the late phase of in vivo biofilm formation. Polyclonal rabbit antibodies raised against SesC reduced the fibrinogen-binding ability of S. epidermidis RP62A and Staphylococcus aureus RN4220 transformants expressing SesC, inhibited in vitro biofilm formation by S. epidermidis strains 10b and 1457, and significantly reduced the numbers of bacteria in a 1-day-old in vivo biofilm (P < 0.001, one-way analysis of variance). Our findings revealed that SesC is a promising target for prevention and treatment of S. epidermidis biofilms because it affects both the primary attachment and biofilm accumulation phases. The precise role of SesC in biofilm formation remains to be identified.
Shahrooei, Mohammad; Hira, Vishal; Stijlemans, Benoit; Merckx, Rita; Hermans, Peter W. M.; Van Eldere, Johan
Inhibition of bacterial biofilm formation and swarming motility by a small synthetic cationic peptide.
Biofilms cause up to 80% of infections and are difficult to treat due to their substantial multidrug resistance compared to their planktonic counterparts. Based on the observation that human peptide LL-37 is able to block biofilm formation at concentrations below its MIC, we screened for small peptides with antibiofilm activity and identified novel synthetic cationic peptide 1037 of only 9 amino acids in length. Peptide 1037 had very weak antimicrobial activity, but at 1/30th the MIC the peptide was able to effectively prevent biofilm formation (>50% reduction in cell biomass) by the Gram-negative pathogens Pseudomonas aeruginosa and Burkholderia cenocepacia and Gram-positive Listeria monocytogenes. Using a flow cell system and a widefield fluorescence microscope, 1037 was shown to significantly reduce biofilm formation and lead to cell death in biofilms. Microarray and follow-up studies showed that, in P. aeruginosa, 1037 directly inhibited biofilms by reducing swimming and swarming motilities, stimulating twitching motility, and suppressing the expression of a variety of genes involved in biofilm formation (e.g., PA2204). Comparison of microarray data from cells treated with peptides LL-37 and 1037 enabled the identification of 11 common P. aeruginosa genes that have a role in biofilm formation and are proposed to represent functional targets of these peptides. Peptide 1037 shows promise as a potential therapeutic agent against chronic, recurrent biofilm infections caused by a variety of bacteria. PMID:22354291
de la Fuente-N��ez, C�sar; Korolik, Victoria; Bains, Manjeet; Nguyen, Uyen; Breidenstein, Elena B M; Horsman, Shawn; Lewenza, Shawn; Burrows, Lori; Hancock, Robert E W
Use of MTT assay for determination of the biofilm formation capacity of microorganisms in metalworking fluids.
Biofilm formation is a well-known problem in management of metalworking fluid systems. Due to persistence of microorganisms within biofilms, the reappearance of various species of bacteria, including nontuberculous mycobacteria is often observed after the use of biocides and/or cleaning of delivery systems and replacement of cooling fluid. The aim of this study was to determine the usefulness of the tetrazolium salt assay (MTT assay) for assessing the viability of bacteria in biofilms formed in vitro in fresh and used cutting oils, as well as their susceptibility to antimicrobial biocides. Biofilms were established in the microtiter plate format. The results showed that quantification of formazan, a product of the tetrazolium salt reduction by electron transport system could be used for determination of the propensity of bacteria to form biofilms in these complex media. The use of the assay allows also determination of antimicrobial activity of biocides against biofilms in fresh and used metalworking fluids. Biofilms produced by Gram-negative isolates recovered from field metalworking fluids as well as the wild bacterial communities differed in metabolic activity depending on the type of fresh coolants. The MTT assay has high-throughput potential and can be efficiently used for determination of biofilm-forming capacity of microorganisms from individual machines in metalworking industry. The use of the assay may also guide the selection of the most appropriate biocide to fight these microorganisms. PMID:23515965
Trafny, El?bieta Anna; Lewandowski, Rafa?; Zawistowska-Marciniak, Irena; St?pi?ska, Ma?gorzata
Resveratrol oligomers inhibit biofilm formation of Escherichia coli O157:H7 and Pseudomonas aeruginosa.
Biofilm formation is closely related to bacterial infection and is also a mechanism of antimicrobial resistance. Hence, the antibiofilm approach provides an alternative to an antibiotic strategy. In this study, the antibiofilm activities of resveratrol (1) and five of its oligomers, namely, ?-viniferin (2), suffruticosol A (3), suffruticosol B (4), vitisin A (5), and vitisin B (6), were investigated against enterohemorrhagic Escherichia coli O157:H7 and Pseudomonas aeruginosa PA14. Vitisin B (6), a stilbenoid tetramer, was found to inhibit biofilm formation by the two bacteria the most effectively and at 5 ?g/mL inhibited E. coli O157:H7 biofilm formation by more than 90%. PMID:24456071
Lee, Jin-Hyung; Kim, Yong-Guy; Ryu, Shi Yong; Cho, Moo Hwan; Lee, Jintae
Human cathelicidin LL-37 prevents bacterial biofilm formation.
Human pathogens often colonize their host by the formation of biofilms. These surface-attached aggregates of bacteria are characterized by a self-produced extracellular matrix, which makes them highly resistant towards antibiotic treatment. Their abilities to adhere to abiotic surfaces (e.g., catheters and other medical devices) also makes bacterial biofilm formation a challenge in modern medicine. Antimicrobial peptides have lately been introduced as a potential class of drug molecules for combating severe hospital-acquired infections. One of these peptides, human cathelicidin LL-37, has recently been demonstrated to bridge innate and adaptive host defence, in addition to facilitating a robust antibiofilm effect at sub-inhibitory concentrations. In this review we will discuss the evidence, potential and challenges for LL-37 as a candidate molecule for therapeutic use. PMID:22917247
Jacobsen, Andreas S; Jenssen, H�vard
Heterologous expression of human paraoxonases in Pseudomonas aeruginosa inhibits biofilm formation and decreases antibiotic resistance
Quorum sensing (QS) regulates virulence and biofilm formation in Pseudomonas aeruginosa and other medically relevant bacteria. Human paraoxonases (hPONs) are a family of closely related enzymes with multiple functions,\\u000a including inactivation of the QS signal molecule in P. aeruginosa. However, there is no direct evidence to show the functions of hPONs on biofilm formation and antibiotic resistance in P. aeruginosa.
Fang Ma; Yao Wang; Yong Zhang; Ning Xiong; Baoyu Yang; Shiyun Chen
Investigation of biofilm formation in clinical isolates of Staphylococcus aureus.
As with many other bacterial species, the most commonly used method to assess staphylococcal biofilm formation in vitro is the microtiter plate assay. This assay is particularly useful for comparison of multiple strains including large-scale screens of mutant libraries. When such screens are applied to the coagulase-negative staphylococci in general, and Staphylococcus epidermidis in particular, they are relatively straightforward by comparison with microtiter plate assays used to assess biofilm formation in other bacterial species. However, in the case of clinical isolates of Staphylococcus aureus, including methicillin-resistant S. aureus, we have found it necessary to employ specific modifications including precoating of the wells of the microtiter plate with plasma proteins and supplementation of the medium with both salt and glucose. In this chapter, we describe the microtiter plate assay in the specific context of clinical isolates of S. aureus and the use of these modifications. A second in vitro method, which also is generally dependent on coating with plasma proteins and supplementation of the growth medium, is the use of flow cells. In this method, bacteria are allowed to attach to a surface and then monitored with respect to their ability to remain attached to the substrate and differentiate into mature biofilms under the constant pressure of fluid shear force. Although flow cells are not applicable to large-scale screens, we have found that they provide a more reproducible and accurate assessment of the capacity of S. aureus clinical isolates to form a biofilm. They also provide a means of analyzing structural differences in biofilm architecture and isolating bacteria and/or spent media for analysis of physiological and metabolic changes associated with the adaptive response to growth in a biofilm. While a primary focus of this chapter is on the use of in vitro assays to assess biofilm formation in clinical isolates of S. aureus, it is important to emphasize two additional considerations. First, it has become increasingly evident that biofilm formation in S. epiderimidis and S. aureus is not equivalent. Additionally, to date, most studies with S. aureus have been done with a very limited number of strains, almost all of which are derived from the NCTC strain designated 8325, and we have found that these strains are not representative of the most relevant clinical isolates. As with the specific elements of our flow cell system, we have written this chapter to reflect our focus on clinical isolates of S. aureus and the specific methods that we have found most reliable in that context. Second, as is often the case, in vitro methods do not necessarily reflect events that occur in vivo. Several in vivo methods to assess biofilm formation have been described, and these generally fall into one of two categories. The first focuses directly on staphylococcal diseases that are generally thought to include a biofilm component (e.g., endocarditis, osteomyelitis, septic arthritis). A discussion of these models is also beyond the scope of this chapter, but examples are easily found in the staphylococcal literature. The second approach uses some form of implanted device in an attempt to focus more directly on implant-associated biofilms. We use a model in which a small piece of Teflon catheter is implanted subcutaneously in mice and used as a substrate for colonization. We have the advantage of using bioluminescent derivatives of S. aureus clinical isolates and the IVIS(R) imaging system. However, because this system is not generally available, we restrict technical comments in this chapter to our use of an implanted catheter model evaluated by direct microbio-logical analysis of explanted catheters (2). PMID:18025674
Cassat, James E; Lee, Chia Y; Smeltzer, Mark S
Investigation of Biofilm Formation in Clinical Isolates of Staphylococcus aureus
Summary As with many other bacterial species, the most commonly used method to assess staphylococcal biofilm formation in vitro is the microtiter plate assay. This assay is particularly useful for comparison of multiple strains including large-scale screens of mutant libraries. When such screens are applied to the coagulase-negative staphylococci in general, and Staphylococcus epidermidis in particular, they are relatively straightforward by comparison with microtiter plate assays used to assess biofilm formation in other bacterial species. However, in the case of clinical isolates of Staphylococcus aureus, including methicillin-resistant S. aureus, we have found it necessary to employ specific modifications including precoating of the wells of the microtiter plate with plasma proteins and supplementation of the medium with both salt and glucose. In this chapter, we describe the microtiter plate assay in the specific context of clinical isolates of S. aureus and the use of these modifications. A second in vitro method, which also is generally dependent on coating with plasma proteins and supplementation of the growth medium, is the use of flow cells. In this method, bacteria are allowed to attach to a surface and then monitored with respect to their ability to remain attached to the substrate and differentiate into mature biofilms under the constant pressure of fluid shear force. Although flow cells are not applicable to large-scale screens, we have found that they provide a more reproducible and accurate assessment of the capacity of S. aureus clinical isolates to form a biofilm. They also provide a means of analyzing structural differences in biofilm architecture and isolating bacteria and/or spent media for analysis of physiological and metabolic changes associated with the adaptive response to growth in a biofilm. While a primary focus of this chapter is on the use of in vitro assays to assess biofilm formation in clinical isolates of S. aureus, it is important to emphasize two additional considerations. First, it has become increasingly evident that biofilm formation in S. epiderimidis and S. aureus is not equivalent. Additionally, to date, most studies with S. aureus have been done with a very limited number of strains, almost all of which are derived from the NCTC strain designated 8325, and we have found that these strains are not representative of the most relevant clinical isolates. As with the specific elements of our flow cell system, we have written this chapter to reflect our focus on clinical isolates of S. aureus and the specific methods that we have found most reliable in that context. Second, as is often the case, in vitro methods do not necessarily reflect events that occur in vivo. Several in vivo methods to assess biofilm formation have been described, and these generally fall into one of two categories. The first focuses directly on staphylococcal diseases that are generally thought to include a biofilm component (e.g., endocarditis, osteomyelitis, septic arthritis). A discussion of these models is also beyond the scope of this chapter, but examples are easily found in the staphylococcal literature. The second approach uses some form of implanted device in an attempt to focus more directly on implant-associated biofilms. We use a model in which a small piece of Teflon catheter is implanted subcutaneously in mice and used as a substrate for colonization. We have the advantage of using bioluminescent derivatives of S. aureus clinical isolates and the IVIS� imaging system. However, because this system is not generally available, we restrict technical comments in this chapter to our use of an implanted catheter model evaluated by direct microbiological analysis of explanted catheters (2).
Cassat, James E.; Lee, Chia Y.; Smeltzer, Mark S.
Clearance of Pseudomonas aeruginosa foreign-body biofilm infections through reduction of the cyclic Di-GMP level in the bacteria.
Opportunistic pathogenic bacteria can engage in biofilm-based infections that evade immune responses and develop into chronic conditions. Because conventional antimicrobials cannot efficiently eradicate biofilms, there is an urgent need to develop alternative measures to combat biofilm infections. It has recently been established that the secondary messenger cyclic diguanosine monophosphate (c-di-GMP) functions as a positive regulator of biofilm formation in several different bacteria. In the present study we investigated whether manipulation of the c-di-GMP level in bacteria potentially can be used for biofilm control in vivo. We constructed a Pseudomonas aeruginosa strain in which a reduction in the c-di-GMP level can be achieved via induction of the Escherichia coli YhjH c-di-GMP phosphodiesterase. Initial experiments showed that induction of yhjH expression led to dispersal of the majority of the bacteria in in vitro-grown P. aeruginosa biofilms. Subsequently, we demonstrated that P. aeruginosa biofilms growing on silicone implants, located in the peritoneal cavity of mice, dispersed after induction of the YhjH protein. Bacteria accumulated temporarily in the spleen after induction of biofilm dispersal, but the mice tolerated the dispersed bacteria well. The present work provides proof of the concept that modulation of the c-di-GMP level in bacteria is a viable strategy for biofilm control. PMID:23690403
Christensen, Louise D; van Gennip, Maria; Rybtke, Morten T; Wu, Hong; Chiang, Wen-Chi; Alhede, Morten; H�iby, Niels; Nielsen, Thomas E; Givskov, Michael; Tolker-Nielsen, Tim
Antimicrobial and antibiofilm activity of quorum sensing peptides and Peptide analogues against oral biofilm bacteria.
Widespread antibiotic resistance is a major incentive for the investigation of novel ways to treat or prevent infections. Much effort has been put into the discovery of peptides in nature accompanied by manipulation of natural peptides to improve activity and decrease toxicity. The ever increasing knowledge about bacteria and the discovery of quorum sensing have presented itself as another mechanism to disrupt the infection process. We have shown that the natural quorum sensing (QS) peptide, competence-stimulating peptide (CSP), used by the caries causing bacteria Streptococcus mutans when used in higher than normally present concentrations can actually contribute to cell death in S. mutans. Using an analogue of this quorum sensing peptide (KBI-3221), we have shown it to be beneficial at decreasing biofilm of various Streptococcus species. This chapter looks at a number of assay methods to test the inhibitory effects of quorum sensing peptides and their analogues on the growth and biofilm formation of oral bacteria. PMID:20094877
LoVetri, Karen; Madhyastha, Srinivasa
Biofilm formation and interactions of bacterial strains found in wastewater treatment systems.
Biofilm formation and adherence properties of 13 bacterial strains commonly found in wastewater treatment systems were studied in pure and mixed cultures using a crystal violet microtiter plate assay. Four different culture media were used, wastewater, acetate medium, glucose medium and diluted nutrient broth. The medium composition strongly affected biofilm formation. All strains were able to form pure culture biofilms within 24 h in at least one of the tested culture media and three strains were able to form biofilm in all four culture media, namely Acinetobacter calcoaceticus ATCC 23055, Comamonas denitrificans 123 and Pseudomonas aeruginosa MBL 0199. The adherence properties assessed were initial adherence, cell surface hydrophobicity, and production of amyloid fibers and extracellular polymeric substances. The growth of dual-strain biofilms showed that five organisms formed biofilm with all 13 strains while seven formed no or only weak biofilm when cocultured. In dual-strain cultures, strains with different properties were able to complement each other, giving synergistic effects. Strongest biofilm formation was observed when a mixture of all 13 bacteria were grown together. These results on attachment and biofilm formation can serve as a tool for the design of tailored systems for the degradation of municipal and industrial wastewater. PMID:18422628
Andersson, Sofia; Kuttuva Rajarao, Gunaratna; Land, Carl Johan; Dalhammar, Gunnel
Hydrogen Peroxide Linked to Lysine Oxidase Activity Facilitates Biofilm Differentiation and Dispersal in Several Gram-Negative Bacteria?
The marine bacterium Pseudoalteromonas tunicata produces an antibacterial and autolytic protein, AlpP, which causes death of a subpopulation of cells during biofilm formation and mediates differentiation, dispersal, and phenotypic variation among dispersal cells. The AlpP homologue (LodA) in the marine bacterium Marinomonas mediterranea was recently identified as a lysine oxidase which mediates cell death through the production of hydrogen peroxide. Here we show that AlpP in P. tunicata also acts as a lysine oxidase and that the hydrogen peroxide generated is responsible for cell death within microcolonies during biofilm development in both M. mediterranea and P. tunicata. LodA-mediated biofilm cell death is shown to be linked to the generation of phenotypic variation in growth and biofilm formation among M. mediterranea biofilm dispersal cells. Moreover, AlpP homologues also occur in several other gram-negative bacteria from diverse environments. Our results show that subpopulations of cells in microcolonies also die during biofilm formation in two of these organisms, Chromobacterium violaceum and Caulobacter crescentus. In all organisms, hydrogen peroxide was implicated in biofilm cell death, because it could be detected at the same time as the killing occurred, and the addition of catalase significantly reduced biofilm killing. In C. violaceum the AlpP-homologue was clearly linked to biofilm cell death events since an isogenic mutant (CVMUR1) does not undergo biofilm cell death. We propose that biofilm killing through hydrogen peroxide can be linked to AlpP homologue activity and plays an important role in dispersal and colonization across a range of gram-negative bacteria.
Mai-Prochnow, Anne; Lucas-Elio, Patricia; Egan, Suhelen; Thomas, Torsten; Webb, Jeremy S.; Sanchez-Amat, Antonio; Kjelleberg, Staffan
A master regulator for biofilm formation by Bacillus subtilis.
Wild strains of Bacillus subtilis are capable of forming architecturally complex communities of cells known as biofilms. Critical to biofilm formation is the eps operon, which is believed to be responsible for the biosynthesis of an exopolysaccharide that binds chains of cells together in bundles. We report that transcription of eps is under the negative regulation of SinR, a repressor that was found to bind to multiple sites in the regulatory region of the operon. Mutations in sinR bypassed the requirement in biofilm formation of two genes of unknown function, ylbF and ymcA, and sinI, which is known to encode an antagonist of SinR. We propose that these genes are members of a pathway that is responsible for counteracting SinR-mediated repression. We further propose that SinR is a master regulator that governs the transition between a planktonic state in which the bacteria swim as single cells in liquid or swarm in small groups over surfaces, and a sessile state in which the bacteria adhere to each other to form bundled chains and assemble into multicellular communities. PMID:15661000
Kearns, Daniel B; Chu, Frances; Branda, Steven S; Kolter, Roberto; Losick, Richard
The inhibitory effects of mushroom extracts on sucrose-dependent oral biofilm formation.
Mushrooms contain large quantities of alpha-glucans. Shiitake (Lentinula edodes), Japan's most popular edible mushroom, has been reported to contain about 6% (weight/dried weight) of alpha-(1,3)-glucan. This glucan is one of the major components of oral biofilm formed by the cariogenic bacteria Streptococcus mutans and Streptococcus sobrinus. We found that extracts from shiitake and other edible mushrooms could reduce preformed biofilms of S. mutans and S. sobrinus in the presence of dextranase. We also investigated the alpha-glucanase activities of shiitake mushroom extracts and their effects on biofilm formation. The extracts possessed alpha-glucanase activity and degraded water-insoluble glucans from mutans streptococci. The extracts strongly inhibited the sucrose-dependent formation of biofilms by S. mutans and S. sobrinus in the presence of dextranase. Our results suggest that some components of mushrooms, including alpha-glucanases, might inhibit the sucrose-induced formation of oral biofilms. PMID:19902205
Yano, Akira; Kikuchi, Sayaka; Yamashita, Yoshihisa; Sakamoto, Yuichi; Nakagawa, Yuko; Yoshida, Yasuo
Quorum sensing and biofilm formation investigated using laser-trapped bacterial arrays
NASA Astrophysics Data System (ADS)
Studies of individual, free-swimming (planktonic) bacteria have yielded much information about their genetic and phenotypic characteristics and about ``quorum sensing,'' the autoinducing process by which bacteria detect high concentrations of other bacteria. However, in most environments the majority of bacteria are not in the planktonic form but are rather in biofilms, which are highly-structured, dynamic communities of multiple bacteria that adhere to a surface and to each other using an extracellular polysaccharide matrix. Bacteria in biofilms are phenotypically very different from their genetically-identical planktonic counterparts. Among other characteristics, they are much more antibiotic-resistant and virulent. Such biofilms form persistent infections on medical implants and in the lungs of cystic fibrosis patients, where Pseudomonas aeruginosa biofilms are the leading cause of lung damage and, ultimately, death. To understand the importance of different extracellular materials, motility mechanisms, and quorum sensing for biofilm formation and stability, we use single-gene knockout mutants and an infrared laser trap to create a bacterial aggregate that serves as a model biofilm and allows us to measure the importance of these factors as a function of trapping time, surface, and nutritional environment.
Gordon, Vernita; Butler, John; Smalyukh, Ivan; Parsek, Matthew; Wong, Gerard
Identification of Small Molecules That Antagonize Diguanylate Cyclase Enzymes To Inhibit Biofilm Formation
Bacterial biofilm formation is responsible for numerous chronic infections, causing a severe health burden. Many of these infections cannot be resolved, as bacteria in biofilms are resistant to the host's immune defenses and antibiotic therapy. New strategies to treat biofilm-based infections are critically needed. Cyclic di-GMP (c-di-GMP) is a widely conserved second-messenger signal essential for biofilm formation. As this signaling system is found only in bacteria, it is an attractive target for the development of new antibiofilm interventions. Here, we describe the results of a high-throughput screen to identify small-molecule inhibitors of diguanylate cyclase (DGC) enzymes that synthesize c-di-GMP. We report seven small molecules that antagonize these enzymes and inhibit biofilm formation by Vibrio cholerae. Moreover, two of these compounds significantly reduce the total concentration of c-di-GMP in V. cholerae, one of which also inhibits biofilm formation by Pseudomonas aeruginosa in a continuous-flow system. These molecules represent the first compounds described that are able to inhibit DGC activity to prevent biofilm formation.
Sambanthamoorthy, Karthik; Sloup, Rudolph E.; Parashar, Vijay; Smith, Joshua M.; Kim, Eric E.; Semmelhack, Martin F.; Neiditch, Matthew B.
Dynamics of Aerial Tower Formation in Bacillus subtilis Biofilms
NASA Astrophysics Data System (ADS)
Biofilms are highly-organized colonies of bacteria that form on surfaces. These colonies form sophisticated structures which make them robust and difficult to remove from environments such as catheters, where they pose serious infection problems. Previous work has shown that sub-mm sized aerial towers form on the surface of Bacillus subtilis colony biofilms. Spore-formation is located preferentially at the tops of these towers, known as fruiting bodies, which aid in the dispersal and propagation of the colony to new sites. The formation of towers is strongly affected by the quorum-sensing molecule surfactin and the cannibalism pathway of the bacteria. In the present work, we use confocal fluorescence microscopy to study the development of individual fruiting bodies, allowing us to visualize the time-dependent spatial distribution of matrix-forming and sporulating bacteria within the towers. With this information, we investigate the physical mechanisms, such as surface tension and polymer concentration gradients, that drive the formation of these structures.
Sinha, Naveen; Seminara, Agnese; Wilking, James; Brenner, Michael; Weitz, Dave
Biofilm Streamer Formation in a Porous Microfluidic Device
NASA Astrophysics Data System (ADS)
Biofilm formation in porous media is of significant importance in many environmental and industrial processes such as bioremediation, oil recovery, and wastewater treatment. In the present study, we fabricated a porous media mimic inside a microfluidic device to observe the growth of bacteria in a porous environment. Here, we report the formation of filamentous structures between the porous structures which are known as streamers. Streamers are made from Polymeric Substance (EPS) and are tethered at one or both ends to a surface, while the rest of the structure floats in the aqueous media. We studied evolution of streamers in different flow rates and identified a tangible link between hydrodynamic conditions and development of these filamentous structures. Our results show that hydrodynamic conditions not only play a key role in determining the formation and stability of the streamers, but also influence their morphology and distribution. These observations, which reveal salient features of biofilm formation in porous media, could open up new avenues for understanding biofilm dynamics in complex natural conditions.
Valiei, Amin
Organic compounds inhibiting S. epidermidis adhesion and biofilm formation
The formation of biofilms on surfaces of indwelling medical devices is a serious medical problem. Staphylococcus epidermidis is a common pathogen found to colonize implanted devices and as a biofilm is more resistant to the host immune system as well as to antibiotic treatments. Combating S. epidermidis infections by preventing or eradicating biofilm formation of the bacterium is therefore a
Zhiqiang Qin; Jingdong Zhang; Yifan Hu; Qijin Chi; Ninell P. Mortensen; Di Qu; S�ren Molin; Jens Ulstrup
Brominated Furanones Inhibit Biofilm Formation by Salmonella enterica Serovar Typhimurium
Salmonella enterica serovar Typhimurium is a main cause of bacterial food-borne diseases. As Salmonella can form biofilms in which it is better protected against antimicrobial agents on a wide diversity of surfaces, it is of interest to explore ways to inhibit biofilm formation. Brominated furanones, originally extracted from the marine alga Delisea pulchra, are known to interfere with biofilm formation
Joost C. A. Janssens; Hans Steenackers; Stijn Robijns; Edith Gellens; Jeremy Levin; Hui Zhao; Kim Hermans; David De Coster; Tine L. Verhoeven; Kathleen Marchal; Jos Vanderleyden; D. E. De Vos; S. C. J. De Keersmaecker
Biofilm formation and the survival of Salmonella Typhimurium on parsley
Although several studies provide evidence that the formation of biofilms by human pathogens on plant tissue is possible, to date there is no direct evidence that biofilms enhance the resistance of plant-associated pathogens to disinfectants or biocides. We hypothesized that biofilm formation would enhance the adhesion and survival of Salmonella on leafy vegetables. To test our hypothesis, we compared the
Anat Lapidot; Ute Romling; Sima Yaron
Bile acids stimulate biofilm formation in Vibrio cholerae.
Vibrio cholerae is a Gram-negative bacterium that causes the acute diarrhoeal disease cholera. After the bacterium is ingested, it passes through the digestive tract, encountering various environmental stresses including the acidic milieu of the stomach and the toxic effects of bile in the duodenum. While these stresses serve as part of a host defence system, V. cholerae has evolved resistance mechanisms that allow it to evade these defences and establish infection. We examined the expression profiles of V. cholerae in response to bile or bile acids and found an induction of biofilm genes. We found that V. cholerae shows significantly enhanced biofilm formation in response to bile acids, and that bacteria within the biofilm are more resistant to the toxicity of bile acids compared with planktonic cells. Bile acid induction of biofilms was found to be dependent on the vps genes (Vibrio polysaccharide synthesis) and their transcriptional activator VpsR, but VpsT is not required. These results contribute to the developing picture of a complex relationship between V. cholerae and its environment within the host during infection. PMID:16359328
Hung, Deborah T; Zhu, Jun; Sturtevant, Derek; Mekalanos, John J
Investigation of N-acyl homoserine lactone (AHL) molecule production in Gram-negative bacteria isolated from cooling tower water and biofilm samples.
In this study, 99 Gram-negative rod bacteria were isolated from cooling tower water, and biofilm samples were examined for cell-to-cell signaling systems, N-acyl homoserine lactone (AHL) signal molecule types, and biofilm formation capacity. Four of 39 (10 %) strains isolated from water samples and 14 of 60 (23 %) strains isolated from biofilm samples were found to be producing a variety of AHL signal molecules. It was determined that the AHL signal molecule production ability and the biofilm formation capacity of sessile bacteria is higher than planktonic bacteria, and there was a statistically significant difference between the AHL signal molecule production of these two groups (p < 0.05). In addition, it was found that bacteria belonging to the same species isolated from cooling tower water and biofilm samples produced different types of AHL signal molecules and that there were different types of AHL signal molecules in an AHL extract of bacteria. In the present study, it was observed that different isolates of the same strains did not produce the same AHLs or did not produce AHL molecules, and bacteria known as AHL producers did not produce AHL. These findings suggest that detection of signal molecules in bacteria isolated from cooling towers may contribute to prevention of biofilm formation, elimination of communication among bacteria in water systems, and blockage of quorum-sensing controlled virulence of these bacteria. PMID:23250628
Haslan, Ezgi; Kimiran-Erdem, Ayten
2D Model for Diffusion of Oxygen with Biochemical Reaction During Biofilm Formation Process in Static Aqueous Medium
NASA Astrophysics Data System (ADS)
A 2D model that describes diffusion of oxygen with biochemical reaction during biofilm formation process in static aqueous medium is presented. The analysis is based on X60 steel placed at the bottom of a container containing produced water inoculated with Leptothrix discophora (iron-oxidizing bacteria). These bacteria form biofilms on the exposed surfaces of the metal. The biofilm-microorganisms absorb oxygen from the produced water through biochemical reaction, resulting in transfer of oxygen from the bulk liquid phase to the biofilm. Predictions of the model are compared with experimental data and good agreement is obtained.
Puyate, Y. T.; Rim-Rukeh, A.
Flagellar Motility Is Critical for Listeria monocytogenes Biofilm Formation
The food-borne pathogen Listeria monocytogenes attaches to environmental surfaces and forms biofilms that can be a source of food contamination, yet little is known about the molecular mechanisms of its biofilm development. We observed that nonmotile mutants were defective in biofilm formation. To investigate how flagella might function during biofilm formation, we compared the wild type with flagellum-minus and paralyzed-flagellum
Katherine P. Lemon; Darren E. Higgins; Roberto Kolter
Observations on biofilm formation in industrial air-cooling units
Observations on biofilm formation in industrial air-cooling units were made over a 60-day operational period. Methods employed included: epifluorescent direct counts of water and slime samples, enumeration of culturable bacteria in water and slime samples, and ultrastructural observations of microbial attachment to formvar coated grids and epoxy resin blocks. Acridine orange direct counts and culturable counts of bacteria in water samples remained constant over the 60-day cycle, while culturable counts in slime samples increased with time. Interfering fluorescent materials present in the slime made accurate direct counts difficult to obtain. Initial increases in numbers of bacteria on formvar coated grids and culturable counts of slime samples were positively correlated with time. However, after 14 days, the formvar deteriorated and direct transmission electron microscopic bacterial counts could no longer be obtained. Submersion of epoxy resin blocks, especially those with pitted surfaces, provided an excellent method for the observation of bacterial attachment and colonization. 21 references, 3 figures.
Liebert, C.A.; Hood, M.A.; Winter, P.A.; Singleton, F.L.
Action of antimicrobial substances produced by different oil reservoir Bacillus strains against biofilm formation.
Microbial colonization of petroleum industry systems takes place through the formation of biofilms, and can result in biodeterioration of the metal surfaces. In a previous study, two oil reservoir Bacillus strains (Bacillus licheniformis T6-5 and Bacillus firmus H(2)O-1) were shown to produce antimicrobial substances (AMS) active against different Bacillus strains and a consortium of sulfate-reducing bacteria (SRB) on solid medium. However, neither their ability to form biofilms nor the effect of the AMS on biofilm formation was adequately addressed. Therefore, here, we report that three Bacillus strains (Bacillus pumilus LF4 -- used as an indicator strain, B. licheniformis T6-5, and B. firmus H(2)O-1), and an oil reservoir SRB consortium (T6lab) were grown as biofilms on glass surfaces. The AMS produced by strains T6-5 and H(2)O-1 prevented the formation of B. pumilus LF4 biofilm and also eliminated pre-established LF4 biofilm. In addition, the presence of AMS produced by H(2)O-1 reduced the viability and attachment of the SRB consortium biofilm by an order of magnitude. Our results suggest that the AMS produced by Bacillus strains T6-5 and H(2)O-1 may have a potential for pipeline-cleaning technologies to inhibit biofilm formation and consequently reduce biocorrosion. PMID:18330565
Korenblum, E; Sebasti�n, G V; Paiva, M M; Coutinho, C M L M; Magalh�es, F C M; Peyton, B M; Seldin, L
Filaments in curved streamlines: rapid formation of Staphylococcus aureus biofilm streamers
NASA Astrophysics Data System (ADS)
Biofilms are surface-associated conglomerates of bacteria that are highly resistant to antibiotics. These bacterial communities can cause chronic infections in humans by colonizing, for example, medical implants, heart valves, or lungs. Staphylococcus aureus, a notorious human pathogen, causes some of the most common biofilm-related infections. Despite the clinical importance of S. aureus biofilms, it remains mostly unknown how physical effects, in particular flow, and surface structure influence biofilm dynamics. Here we use model microfluidic systems to investigate how environmental factors, such as surface geometry, surface chemistry, and fluid flow affect biofilm development of S. aureus. We discovered that S. aureus rapidly forms flow-induced, filamentous biofilm streamers, and furthermore if surfaces are coated with human blood plasma, streamers appear within minutes and clog the channels more rapidly than if the channels are uncoated. To understand how biofilm streamer filaments reorient in flows with curved streamlines to bridge the distances between corners, we developed a mathematical model based on resistive force theory of slender filaments. Understanding physical aspects of biofilm formation of S. aureus may lead to new approaches for interrupting biofilm formation of this pathogen.
Kim, Minyoung Kevin; Drescher, Knut; Pak, On Shun; Bassler, Bonnie L.; Stone, Howard A.
Biofilm formation at warming temperature: acceleration of microbial colonization and microbial interactive effects
River biofilms that grow on wet benthic surface are mainly composed of bacteria, algae, cyanobacteria and protozoa embedded in a polysaccharide matrix. The effects of increased river water temperature on biofilm formation were investigated. A laboratory experiment was designed employing two temperatures (11.1�13.2�C, night�day; 14.7�16.0�C, night�day) and two nutrient levels (0.054 mg P l, 0.75 mg N l; 0.54 mg
Ver�nica D�az Villanueva; Thomas Schwartz; Anna M. Roman�
NspS, a Predicted Polyamine Sensor, Mediates Activation of Vibrio cholerae Biofilm Formation by Norspermidine
Vibrio cholerae is both an environmental bacterium and a human intestinal pathogen. The attachment of bacteria to surfaces in biofilms is thought to be an important feature of the survival of this bacterium both in the environment and within the human host. Biofilm formation occurs when cell-surface and cell-cell contacts are formed to make a three-dimensional structure characterized by pillars
Ece Karatan; Tammi R. Duncan; Paula I. Watnick
Manganese Ion Increases LAB-yeast Mixed-species Biofilm Formation
Remarkable LAB-yeast mixed-species biofilm was formed by lactic acid bacteria (LAB) Lactobacillus plantarum ML11-11 isolated from Fukuyama pot vinegar and Saccharomyces cerevisiae. This mixed-species biofilm formation increased in proportion to the YPD medium concentration but decreased in proportion to the MRS medium concentration. The effect of MRS components on mixed-species biofilm formation was investigated in a YPD medium environment, and it was clarified that beef extract (one of the MRS medium components) decreased mixed-species biofilm formation. On the other hand, manganese sulfate (another component in MRS) remarkably increased both LAB single- and LAB-yeast mixed-species biofilm formation. LAB single- and mixed-species biofilm formation were increased in proportion to the manganese sulfate concentration up to 1?mM and 100 ?M, respectively. The growth of L. plantarum ML11-11 was increased significantly by the addition of 10 ?M manganese sulfate and was resistant to higher concentration of up to 100?mM, but growth of S. cerevisiae was sensitive to manganese ion above 100 ?M. These results suggested that mixed-species biofilm formation could be controlled artificially by controlling the manganese ion level.
NOZAKA, Soma; FURUKAWA, Soichi; SASAKI, Miwa; HIRAYAMA, Satoru; OGIHARA, Hirokazu; MORINAGA, Yasushi
[Sliding motility and biofilm formation in mycobacteria].
Using as a model Mycobacterium smegmatis, a non-motile microorganism, we have studied for the first time in mycobacteria the phenomenon known as sliding motility, as well as the process of biofilm formation. A screen of random transposon mutants was performed in order to identify the genes required for mycobacterial sliding over the surface of motility plates. The genetic analysis described here has been published recently. The genes required for sliding and for biofilm formation (mps and tmtpC) are involved in the biosynthesis of the glycopeptidolipids (GPLs) and their transport to the mycobacterial capsule. Based on our results, we suggest a model for the role of the GPLs in both phenomena. PMID:11899706
Recht, J; Mart�nez, A; Torello, S; Kolter, R
Electroactive mixed culture biofilms in microbial bioelectrochemical systems: The role of temperature for biofilm formation and performance
In this paper we investigate the temperature dependence and temperature limits of waste water derived anodic microbial biofilms. We demonstrate that these biofilms are active in a temperature range between 5�C and 45�C. Elevated temperatures during initial biofilm growth not only accelerate the biofilm formation process, they also influence the bioelectrocatalytic performance of these biofilms when measured at identical operation
Sunil A. Patil; Falk Harnisch; Balasaheb Kapadnis; Uwe Schr�der
[Biofilms of pathogenic bacteria and their role in chronization of infectious process. The search for the means to control biofilms].
The role of biofilms in the environment and in the hosts of pathogenic bacteria has recently attracted much attention of researchers. Microorganisms form biofilms at any biotic and abiotic surfaces and thereby cause serious problems in medical practice and other areas of human activity. Biofilms have been shown to be pathogenetic factors responsible for chronization of infectious process. The data are presented illustrating ubiquitous nature of biofilms, their structural and functional characteristics, and modern methods for the study of microbial communities. The discussion is focused on the role of biofilms in chronization of infectious process, enhanced resistance of biofilm organisms to antibiotics and its underlying mechanisms. Approaches to the search for new means for biofilm control during chronic infections are considered. PMID:22168037
Romanova, Iu M; Didenko, L V; Tolordava, � R; Gintsburg, A L
Gene expression during S. epidermidis biofilm formation on biomaterials.
Biomaterial-centered infections are initiated by adhesion of bacteria to an implant, followed by colonization and mature biofilm formation. Staphylococcus epidermidis is commonly identified as the cause of these device-centered infections. This study used an in vitro model to evaluate temporal changes in the expression of genes-icaADBC, agrBDCA, aap, and atle-that have been identified to play a role in the pathogenesis of S. epidermidis infections. Real-time reverse transcription-polymerase chain reaction was used to determine changes in gene expression from S epidermidis biofilm grown on polyurethanes (Elasthane 80A, hydrophobic) modified with polyethylene oxide (Elasthane 80A-6PEO, hydrophilic) and fluorocarbon (Elasthane 80A-6F, hydrophobic). In vitro expression of the ica locus, which is involved in initial adhesion and intracellular aggregation, increased up to 100-fold from 2 to 48 h, whereas gene expression for autolysin AtlE decreased slightly from 2 to 12 h, followed by a 10-fold increase by 48 h. Upregulation of the aap gene associated with bacterial accumulation and the agr quorum-sensing system was observed during biofilm formation over 48 h. In addition, no correlation was observed between S. epidermidis gene expression and biomaterial surface chemistry. This study used an in vitro model to demonstrate that enhanced expression of the atle, aap, agr, and ica genes plays an important role in initial foreign body colonization and potentially in the establishment of a device-associated infection. PMID:22623350
Patel, Jasmine D; Colton, Erica; Ebert, Michael; Anderson, James M
Biofilm formation of Clostridium perfringens and its exposure to low-dose antimicrobials.
Clostridium perfringens is an opportunistic pathogen that can cause food poisoning in humans and various enterotoxemia in animal species. Very little is known on the biofilm of C. perfringens and its exposure to subminimal inhibitory concentrations of antimicrobials. This study was undertaken to address these issues. Most of the C. perfringens human and animal isolates tested in this study were able to form biofilm (230/277). Porcine clinical isolates formed significantly more biofilm than the porcine commensal isolates. A subgroup of clinical and commensal C. perfringens isolates was randomly selected for further characterization. Biofilm was found to protect C. perfringens bacterial cells from exposure to high concentrations of tested antimicrobials. Exposure to low doses of some of these antimicrobials tended to lead to a diminution of the biofilm formed. However, a few isolates showed an increase in biofilm formation when exposed to low doses of tylosin, bacitracin, virginiamycin, and monensin. Six isolates were randomly selected for biofilm analysis using scanning laser confocal microscopy. Of those, four produced more biofilm in presence of low doses of bacitracin whereas biofilms formed without bacitracin were thinner and less elevated. An increase in the area occupied by bacteria in the biofilm following exposure to low doses of bacitracin was also observed in the majority of isolates. Morphology examination revealed flat biofilms with the exception of one isolate that demonstrated a mushroom-like biofilm. Matrix composition analysis showed the presence of proteins, beta-1,4 linked polysaccharides and extracellular DNA, but no poly-beta-1,6-N-acetyl-D-glucosamine. This study brings new information on the biofilm produced by C. perfringens and its exposure to low doses of antimicrobials. PMID:24795711
Charlebois, Audrey; Jacques, Mario; Archambault, Marie
Biofilm formation of Clostridium perfringens and its exposure to low-dose antimicrobials
Clostridium perfringens is an opportunistic pathogen that can cause food poisoning in humans and various enterotoxemia in animal species. Very little is known on the biofilm of C. perfringens and its exposure to subminimal inhibitory concentrations of antimicrobials. This study was undertaken to address these issues. Most of the C. perfringens human and animal isolates tested in this study were able to form biofilm (230/277). Porcine clinical isolates formed significantly more biofilm than the porcine commensal isolates. A subgroup of clinical and commensal C. perfringens isolates was randomly selected for further characterization. Biofilm was found to protect C. perfringens bacterial cells from exposure to high concentrations of tested antimicrobials. Exposure to low doses of some of these antimicrobials tended to lead to a diminution of the biofilm formed. However, a few isolates showed an increase in biofilm formation when exposed to low doses of tylosin, bacitracin, virginiamycin, and monensin. Six isolates were randomly selected for biofilm analysis using scanning laser confocal microscopy. Of those, four produced more biofilm in presence of low doses of bacitracin whereas biofilms formed without bacitracin were thinner and less elevated. An increase in the area occupied by bacteria in the biofilm following exposure to low doses of bacitracin was also observed in the majority of isolates. Morphology examination revealed flat biofilms with the exception of one isolate that demonstrated a mushroom-like biofilm. Matrix composition analysis showed the presence of proteins, beta-1,4 linked polysaccharides and extracellular DNA, but no poly-beta-1,6-N-acetyl-D-glucosamine. This study brings new information on the biofilm produced by C. perfringens and its exposure to low doses of antimicrobials.
Charlebois, Audrey; Jacques, Mario; Archambault, Marie
Effects of growth conditions on biofilm formation by Actinobacillus pleuropneumoniae.
Biofilm formation is an important virulence trait of many bacterial pathogens. It has been reported in the literature that only two of the reference strains of the swine pathogen Actinobacillus pleuropneumoniae, representing serotypes 5b and 11, were able to form biofilm in vitro. In this study, we compared biofilm formation by the serotype 1 reference strain S4074 of A. pleuropneumoniae grown in five different culture media. We observed that strain S4074 of A. pleuropneumoniae is able to form biofilms after growth in one of the culture conditions tested brain heart infusion (BHI medium, supplier B). Confocal laser scanning microscopy using a fluorescent probe specific to the poly-N-acetylglucosamine (PGA) polysaccharide further confirmed biofilm formation. In accordance, biofilm formation was susceptible to dispersin B, a PGA hydrolase. Transcriptional profiles of A. pleuropneumoniae S4074 following growth in BHI-B, which allowed a robust biofilm formation, and in BHI-A, in which only a slight biofilm formation was observed, were compared. Genes such as tadC, tadD, genes with homology to autotransporter adhesins as well as genes pgaABC involved in PGA biosynthesis and genes involved in zinc transport were up-regulated after growth in BHI-B. Interestingly, biofilm formation was inhibited by zinc, which was found to be more present in BHI-A (no or slight biofilm) than in BHI-B. We also observed biofilm formation in reference strains representing serotypes 3, 4, 5a, 12 and 14 as well as in 20 of the 37 fresh field isolates tested. Our data indicate that A. pleuropneumoniae has the ability to form biofilms under appropriate growth conditions and transition from a biofilm-positive to a biofilm-negative phenotype was reversible. PMID:19737507
Labrie, Jos�e; Pelletier-Jacques, Genevi�ve; Deslandes, Vincent; Ramjeet, Mahendrasingh; Auger, Eliane; Nash, John H E; Jacques, Mario
Biomaterials surfaces capable of resisting fungal attachment and biofilm formation.
Microbial attachment onto biomedical devices and implants leads to biofilm formation and infection; such biofilms can be bacterial, fungal, or mixed. In the past 15 years, there has been an increasing research effort into antimicrobial surfaces but the great majority of these publications present research on bacteria, with some reports also testing resistance to fungi. Very few studies have focused exclusively on antifungal surfaces. However, with increasing recognition of the importance of fungal infections to human health, particularly related to infections at biomaterials, it would seem that the interest in antifungal surfaces is disproportionately low. In studies of both bacteria and fungi, fungi tend to be the minor focus with hypothesized antibacterial mechanisms of action often generalized to also explain the antifungal effect. Yet bacteria and fungi represent two Distinct biological Domains and possess substantially different cellular physiology and structure. Thus it is questionable whether these generalizations are valid. Here we review the scientific literature focusing on surface coatings prepared with antifungal agents covalently attached to the biomaterial surface. We present a critical analysis of generalizations and their evidence. This review should be of interest to researchers of "antimicrobial" surfaces by addressing specific issues that are key to designing and understanding antifungal biomaterials surfaces and their putative mechanisms of action. PMID:24211473
Coad, Bryan R; Kidd, Sarah E; Ellis, David H; Griesser, Hans J
Involvement of Mycobacterium smegmatis undecaprenyl phosphokinase in biofilm and smegma formation.
We describe a Mycobacterium smegmatis mutant with impaired biofilm and smegma formation. A gene homologous to Escherichia coli bacA, which has been proposed to play a role as undecaprenyl phosphokinase (Upk) was unmarked in-frame deleted from M. smegmatis. Though Upk is involved in cell wall synthesis, the surface of the mutant strain appeared virtually comparable to that of the wild type by electron microscopy. The absence of Upk influenced colony morphology and bacitracin resistance. The M. smegmatis Deltaupk mutant developed a biofilm characterized by scattered islands of bacteria distinct from the completely covered biofilm surface observed for wild-type bacteria. We further demonstrate biological consequences of upk deletion for smegma development in an in vivo model. These results suggest the upk gene to be essential in biofilm and smegma development. PMID:15345226
R�se, Lars; Kaufmann, Stefan H E; Daugelat, Sabine
inhibitory effects of citral, cinnamaldehyde, and tea polyphenols on mixed biofilm formation by foodborne Staphylococcus aureus and Salmonella enteritidis.
Biofilms are significant hazards in the food industry. In this study, we investigated the effects of food additive such as citral, cinnamaldehyde, and tea polyphenols on mixed biofilm formation by foodborne Staphylococcus aureus and Salmonella serotype Enteritidis. The adhesion rates of mixed strains in sub-MIC of additives were determined by a microtiter plate assay and bacterial communication signal autoinducer 2 (AI-2) production via a bioluminescence reporter Vibrio harveyi BB170. The structure of mixed biofilm was analyzed using scanning electron microscopy. The effect of the disinfectants hydrogen peroxide, sodium hypochlorite, and peracetic acid was tested on the mixed biofilm. Our results demonstrated that citral, cinnamaldehyde, and tea polyphenols were able to significantly inhibit mixed biofilm formation, while citral could reduce the synthesis of AI-2. Conversely, we observed a significant increase in AI-2 mediated by cinnamaldehyde. Tea polyphenols at lower concentrations induced AI-2 synthesis; however, AI-2 synthesis was significantly inhibited at higher concentrations (300 m g/ml). Food additives inhibited the adhesion of mixed bacteria on stainless steel chips and increased the sensitivity of the mixed biofilm to disinfectants. In conclusion, citral, cinnamaldehyde, and tea polyphenols had strong inhibitory effects on mixed biofilm formation and also enhanced the effect of disinfectant on mixed biofilm formation. This study provides a scientific basis for the application of natural food additives to control biofilm formation of foodborne bacteria. PMID:24853514
Zhang, Hongmei; Zhou, Wenyuan; Zhang, Wenyan; Yang, Anlin; Liu, Yanlan; Jiang, Yan; Huang, Shaosong; Su, Jianyu
Multiple Vibrio fischeri genes are involved in biofilm formation and host colonization
Biofilms are increasingly recognized as the predominant form for survival in the environment for most bacteria. The successful colonization of Vibrio fischeri in its squid host Euprymna tasmanica, involves complex microbe-host interactions mediated by specific genes that are essential for biofilm formation and colonization. In the present investigation, structural and regulatory genes were selected to study their role in biofilm formation and host colonization. We have mutated several genes (pilT, pilU, flgF, motY, ibpA and mifB) by an insertional inactivation strategy. Results demonstrate that structural genes responsible for synthesis of type IV pili and flagella are crucial for biofilm formation and host infection. Moreover, regulatory genes affect colony aggregation by various mechanisms including alteration of synthesis of transcriptional factors and regulation of extracellular polysaccharide production. These results reflect the significance of how genetic alterations influence communal behavior, which is important in understanding symbiotic relationships.
Chavez-Dozal, Alba; Hogan, David; Gorman, Clayton; Quintanal-Villalonga, Alvaro; Nishiguchi, Michele K.
New properties of wheat bran: anti-biofilm activity and interference with bacteria quorum-sensing systems.
Some plant extracts, have been demonstrated to interfere with the microbial metabolism of several pathogenic bacteria. Within this antimicrobial properties it has been described the potential to inhibit or destroy biofilms or to interfere in quorum-sensing (QS) systems. However, to our knowledge, no study exploring this potential of wheat-bran (WB) has been published. The purpose of the present study is to evaluate the anti-biofilm activity of WB against a cow mastitis strain of Staphylococcus aureus and also its possible interference with bacterial QS systems. The potential of inhibition and destruction of the biofilm was studied by different in vitro assays. Also, we tested the ability of WB to interfere in bacterial QS by degrading acyl-homoserine lactones (AHL) as one of the most studied QS signal molecules for Gram-negative bacteria. The soluble extract of WB at 0.5% showed anti-biofilm activity, inhibiting biofilm formation and also destroying it. Similarly, the >?300?kDa fraction from WB had significant anti-biofilm activity in both in vitro assays. The WB also showed a potential to interfere with bacterial QS systems, as it was demonstrated to contain certain lactonase activity able to reduce AHL concentration in the medium. The present study reveals two additional beneficial properties of WB extract never explored before, which may be related to the presence of defence compounds in the plant extract able to interfere with microbial biofilms and also QS systems. PMID:24588934
Gonz�lez-Ortiz, Gemma; Quarles Van Ufford, H C; Halkes, S Bart A; Cerd�-Cu�llar, Marta; Beukelman, Cees J; Pieters, Roland J; Liskamp, Rob M J; P�rez, Jos� F; Mart�n-Orue, Susana M
Environmental and genetic factors that contribute to Escherichia coli K-12 biofilm formation
Biofilms are communities of bacteria whose formation on surfaces requires a large portion of the bacteria�s transcriptional network. To identify environmental conditions and transcriptional regulators that contribute to sensing these conditions, we used a high-throughput approach to monitor biofilm biomass produced by an isogenic set of Escherichia coli K-12 strains grown under combinations of environmental conditions. Of the environmental combinationsd, growth in tryptic soy broth at 37�C supported the most biofilm production. To analyze the complex relationships between the diverse cell surface organelles, transcriptional regulators, and metabolic enzymes represented by the tested mutant set, we used a novel vector-item pattern-mining algorithm. The algorithm related biofilm amounts to the functional annotations of each mutated protein. The pattern with the best statistical significance was the gene ontology �pyruvate catabolic process,� which is associated with enzymes of acetate metabolism. Phenotype microarray experiments illustrated that carbon sources that are metabolized to acetyl-coenzyme A, acetyl phosphate, and acetate are particularly supportive of biofilm formation. Scanning electron microscopy revealed structural differences between mutants that lack acetate metabolism enzymes and their parent and confirmed the quantitative differences. We conclude that acetate metabolism functions as a metabolic sensor, transmitting changes in environmental conditions to biofilm biomass and structure.
Pruss, Birgit M.; Verma, Karan; Samanta, Priyankar; Sule, Preeti; Kumar, Sunil; Wu, Jianfei; Christianson, David; Horne, Shelley M.; Stafslien, Shane J.; Wolfe, Alan J.; Denton, Anne
Phytotherapeutic prevention of dental biofilm formation.
The antimicrobial and biofilm formation preventive properties of Mentha piperita and Rosmarinus officinalis essential oils and chlorhexidine were assessed against Streptococcus mutans and Streptococcus pyogenes. 26 and 20 compounds were identified by GC and GC-MS analysis in hydrodistilled oils from M. piperita and R. officinalis, respectively. The minimal bactericidal concentrations (MBC) of the M. piperita and R. officinalis oils and chlorhexidine were (6000, 2000, 8000 ppm) and (1000, 4000, 1000 ppm) for S. mutans and S. pyogenes, respectively. The decimal reduction time (D) of S. mutans exposed to the oils at their MBC levels was 2.8 min while chlorhexidine showed a longer time. The D values of S. pyogenes on exposure to the MBC levels of M. piperita and R. officinalis oils and of chlorhexidine were 2.14, 4.28 and 2.8 min, indicating a higher efficacy of M. piperita oil. Biofilm formation was performed by growing S. mutans culture with and without essential oils in LB medium in polystyrene tubes. In vitro biofilm inhibitory properties were in the order M. piperita > R. officinalis > chlorhexidine. In vivo experiments on the antibiofilm properties revealed that all concentrations of the oils were significantly (p < 0.001) more effective than chlorhexidine. In conclusion, essential oils may be considered as safe agents in the development of novel antibiofilm agents. PMID:18729251
Rasooli, Iraj; Shayegh, Shojaedin; Taghizadeh, Massoud; Astaneh, Shakiba Darvish Alipoor
Rapid evolution of culture-impaired bacteria during adaptation to biofilm growth.
Biofilm growth increases the fitness of bacteria in harsh conditions. However, bacteria from clinical and environmental biofilms can exhibit impaired growth in culture, even when the species involved are readily culturable and permissive conditions are used. Here, we show that culture-impaired variants of Pseudomonas aeruginosa arise rapidly and become abundant in laboratory biofilms. The culture-impaired phenotype is caused by mutations that alter the outer-membrane lipopolysaccharide structure. Within biofilms, the lipopolysaccharide mutations markedly increase bacterial fitness. However, outside the protected biofilm environment, the mutations sensitize the variants to killing by a self-produced antimicrobial agent. Thus, a biofilm-mediated adaptation produces a stark fitness trade-off that compromises bacterial survival in culture. Trade-offs like this could limit the ability of bacteria to transition between biofilm growth and the free-living state and produce bacterial populations that escape detection by culture-based sampling. PMID:24412364
Penterman, Jon; Nguyen, Dao; Anderson, Erin; Staudinger, Benjamin J; Greenberg, Everett P; Lam, Joseph S; Singh, Pradeep K
Inhibitory effect of cyclic trihydroxamate siderophore, desferrioxamine E, on the biofilm formation of Mycobacterium species.
Formation of biofilm in pathogenic bacteria defends them from antibiotics and the immune system of a host's life. Hence, investigation of the molecular mechanisms of biofilm formation and search for new substances counteracting this formation are becoming an attractive research area. In the course of our search for new inhibitors of biofilm formation in Mycobacterium species, we rediscovered a cyclic trihydroxamate siderophore, desferrioxamine E, from the culture of the marine-derived Actinomycete MS67. Desferrioxamine E inhibited biofilm formation of Mycobacterium smegmatis and M. bovis BACILLE de CALMETTE et GU�RIN (BCG) with minimum inhibitory concentration (MIC) value of 10 �M, while no anti-microbial activity was observed up to 160 �M. Desferrioxamine E was also able to restore the anti-microbial activity of isoniazid against M. smegmatis by inhibiting biofilm formation. Mechanistic analysis of desferrioxamine E suggested that such inhibition might come from the depletion of iron in the medium, which is essential for biofilm formation in Mycobacterium species. PMID:21628895
Ishida, Shunsuke; Arai, Masayoshi; Niikawa, Hiroki; Kobayashi, Motomasa
Stringent Response Regulation of Biofilm Formation in Vibrio cholerae
Biofilm formation is a key factor in Vibrio cholerae environmental survival and host colonization. Production of biofilm enables V. cholerae to survive and persist in aquatic environments and aids in the passage through the gastric acid barrier to allow access to the small intestine. The genes involved in biofilm formation are regulated by the transcriptional activators vpsR and vpsT, which are in turn transcriptionally regulated by a number of environmental signals. In this study, the role of the stringent response in biofilm formation was examined. V. cholerae mutants deficient in stringent response had a reduced ability to form biofilms, although they were not completely deficient in biofilm formation. There are three (p)ppGpp synthases in V. cholerae: RelA, SpoT, and RelV. All three synthases were necessary for vpsR transcription, with RelV showing the strongest effect. RelA was the only synthase that was necessary for vpsT expression. Stringent response regulation of vpsR and vpsT was shown to partially occur through rpoS. Biofilm formation in V. cholerae is controlled by a complex regulatory apparatus, with negative regulators of biofilm gene expression, such as quorum sensing, and positive regulators of biofilm genes, including stringent response, interacting to ensure that biofilm formation is coordinated with the environment.
He, Huajun; Cooper, Jennifer N.; Mishra, Arunima
Stringent response regulation of biofilm formation in Vibrio cholerae.
Biofilm formation is a key factor in Vibrio cholerae environmental survival and host colonization. Production of biofilm enables V. cholerae to survive and persist in aquatic environments and aids in the passage through the gastric acid barrier to allow access to the small intestine. The genes involved in biofilm formation are regulated by the transcriptional activators vpsR and vpsT, which are in turn transcriptionally regulated by a number of environmental signals. In this study, the role of the stringent response in biofilm formation was examined. V. cholerae mutants deficient in stringent response had a reduced ability to form biofilms, although they were not completely deficient in biofilm formation. There are three (p)ppGpp synthases in V. cholerae: RelA, SpoT, and RelV. All three synthases were necessary for vpsR transcription, with RelV showing the strongest effect. RelA was the only synthase that was necessary for vpsT expression. Stringent response regulation of vpsR and vpsT was shown to partially occur through rpoS. Biofilm formation in V. cholerae is controlled by a complex regulatory apparatus, with negative regulators of biofilm gene expression, such as quorum sensing, and positive regulators of biofilm genes, including stringent response, interacting to ensure that biofilm formation is coordinated with the environment. PMID:22467780
He, Huajun; Cooper, Jennifer N; Mishra, Arunima; Raskin, David M
Biofilm-forming bacteria with varying tolerance to peracetic acid from a paper machine
Biofilms cause runnability problems in paper machines and are therefore controlled with biocides. Peracetic acid is usually\\u000a effective in preventing bulky biofilms. This study investigated the microbiological status of a paper machine where low concentrations\\u000a (?15�ppm active ingredient) of peracetic acid had been used for several years. The paper machine contained a low amount of\\u000a biofilms. Biofilm-forming bacteria from this
Stiina Rasimus; Marko Kolari; Hannu Rita; Douwe Hoornstra; Mirja Salkinoja-Salonen
Establishment and Early Succession of a Multispecies Biofilm Composed of Soil Bacteria
Most soil bacteria are likely to be organized in biofilms on roots, litter, or soil particles. Studies of such biofilms are\\u000a complicated by the many nonculturable species present in soil, as well as the interspecific bacterial interactions affecting\\u000a biofilm biology. We in this study describe the development of a biofilm flow model and use this system to establish an early
Mette Burm�lle; Lars H. Hansen; S�ren J. S�rensen
Mechanisms and regulation of surface interactions and biofilm formation in Agrobacterium.
For many pathogenic bacteria surface attachment is a required first step during host interactions. Attachment can proceed to invasion of host tissue or cells or to establishment of a multicellular bacterial community known as a biofilm. The transition from a unicellular, often motile, state to a sessile, multicellular, biofilm-associated state is one of the most important developmental decisions for bacteria. Agrobacterium tumefaciens genetically transforms plant cells by transfer and integration of a segment of plasmid-encoded transferred DNA (T-DNA) into the host genome, and has also been a valuable tool for plant geneticists. A. tumefaciens attaches to and forms a complex biofilm on a variety of biotic and abiotic substrates in vitro. Although rarely studied in situ, it is hypothesized that the biofilm state plays an important functional role in the ecology of this organism. Surface attachment, motility, and cell division are coordinated through a complex regulatory network that imparts an unexpected asymmetry to the A. tumefaciens life cycle. In this review, we describe the mechanisms by which A. tumefaciens associates with surfaces, and regulation of this process. We focus on the transition between flagellar-based motility and surface attachment, and on the composition, production, and secretion of multiple extracellular components that contribute to the biofilm matrix. Biofilm formation by A. tumefaciens is linked with virulence both mechanistically and through shared regulatory molecules. We detail our current understanding of these and other regulatory schemes, as well as the internal and external (environmental) cues mediating development of the biofilm state, including the second messenger cyclic-di-GMP, nutrient levels, and the role of the plant host in influencing attachment and biofilm formation. A. tumefaciens is an important model system contributing to our understanding of developmental transitions, bacterial cell biology, and biofilm formation. PMID:24834068
Heindl, Jason E; Wang, Yi; Heckel, Brynn C; Mohari, Bitan; Feirer, Nathan; Fuqua, Clay
Mechanisms and regulation of surface interactions and biofilm formation in Agrobacterium
For many pathogenic bacteria surface attachment is a required first step during host interactions. Attachment can proceed to invasion of host tissue or cells or to establishment of a multicellular bacterial community known as a biofilm. The transition from a unicellular, often motile, state to a sessile, multicellular, biofilm-associated state is one of the most important developmental decisions for bacteria. Agrobacterium tumefaciens genetically transforms plant cells by transfer and integration of a segment of plasmid-encoded transferred DNA (T-DNA) into the host genome, and has also been a valuable tool for plant geneticists. A. tumefaciens attaches to and forms a complex biofilm on a variety of biotic and abiotic substrates in vitro. Although rarely studied in situ, it is hypothesized that the biofilm state plays an important functional role in the ecology of this organism. Surface attachment, motility, and cell division are coordinated through a complex regulatory network that imparts an unexpected asymmetry to the A. tumefaciens life cycle. In this review, we describe the mechanisms by which A. tumefaciens associates with surfaces, and regulation of this process. We focus on the transition between flagellar-based motility and surface attachment, and on the composition, production, and secretion of multiple extracellular components that contribute to the biofilm matrix. Biofilm formation by A. tumefaciens is linked with virulence both mechanistically and through shared regulatory molecules. We detail our current understanding of these and other regulatory schemes, as well as the internal and external (environmental) cues mediating development of the biofilm state, including the second messenger cyclic-di-GMP, nutrient levels, and the role of the plant host in influencing attachment and biofilm formation. A. tumefaciens is an important model system contributing to our understanding of developmental transitions, bacterial cell biology, and biofilm formation.
Heindl, Jason E.; Wang, Yi; Heckel, Brynn C.; Mohari, Bitan; Feirer, Nathan; Fuqua, Clay
Variation in Biofilm Formation among Strains of Listeria monocytogenes
Contamination of food by Listeria monocytogenes is thought to occur most frequently in food-processing environments where cells persist due to their ability to attach to stainless steel and other surfaces. Once attached these cells may produce multicellular biofilms that are resistant to disinfection and from which cells can become detached and contaminate food products. Because there is a correlation between virulence and serotype (and thus phylogenetic division) of L. monocytogenes, it is important to determine if there is a link between biofilm formation and disease incidence for L. monocytogenes. Eighty L. monocytogenes isolates were screened for biofilm formation to determine if there is a robust relationship between biofilm formation, phylogenic division, and persistence in the environment. Statistically significant differences were detected between phylogenetic divisions. Increased biofilm formation was observed in Division II strains (serotypes 1/2a and 1/2c), which are not normally associated with food-borne outbreaks. Differences in biofilm formation were also detected between persistent and nonpersistent strains isolated from bulk milk samples, with persistent strains showing increased biofilm formation relative to nonpersistent strains. There were no significant differences detected among serotypes. Exopolysaccharide production correlated with cell adherence for high-biofilm-producing strains. Scanning electron microscopy showed that a high-biofilm-forming strain produced a dense, three-dimensional structure, whereas a low-biofilm-forming strain produced a thin, patchy biofilm. These data are consistent with data on persistent strains forming biofilms but do not support a consistent relationship between enhanced biofilm formation and disease incidence.
Borucki, Monica K.; Peppin, Jason D.; White, David; Loge, Frank; Call, Douglas R.
Bistability and biofilm formation in Bacillus subtilis.
Biofilms of Bacillus subtilis consist of long chains of cells that are held together in bundles by an extracellular matrix of exopolysaccharide and the protein TasA. The exopolysaccharide is produced by enzymes encoded by the epsA-O operon and the gene encoding TasA is located in the yqxM-sipW-tasA operon. Both operons are under the control of the repressor SinR. Derepression is mediated by the antirepressor SinI, which binds to SinR with a 1:1 stoichiometry. Paradoxically, in medium promoting derepression of the matrix operons, the overall concentration of SinR in the culture greatly exceeded that of SinI. We show that under biofilm-promoting conditions sinI, which is under the control of the response regulator Spo0A, was expressed only in a small subpopulation of cells, whereas sinR was expressed in almost all cells. Activation of Spo0A is known to be subject to a bistable switch, and we infer that SinI reaches levels sufficient to trigger matrix production only in the subpopulation of cells in which Spo0A is active. Additionally, evidence suggests that sinI is expressed at intermediate, but not low or high, levels of Spo0A activity, which may explain why certain nutritional conditions are more effective in promoting biofilm formation than others. PMID:18047568
Chai, Yunrong; Chu, Frances; Kolter, Roberto; Losick, Richard
Influence of attached bacteria and biofilm on double-layer capacitance during biofilm monitoring by electrochemical impedance spectroscopy.
Development of an effective strategy for biofilm control in water-related system has become a matter of significant concern nowadays. Electrochemical monitoring, especially electrochemical impedance spectroscopy (EIS), is one of the efficient approaches to dealing with biofilm-related issues. However, currently used EIS methods without a redox probe intend to detect all effects generated from media components, bacteria, and bacterial metabolites, which used to make the signals from the attached bacteria and biofilm weakened. In this study, we tried improved EIS measurement to monitor bacterial adhesion and biofilm maturation using a double-layer capacitance. In this improved method, we minimized background signal by subtracting the interference of electrolyte caused by bacterial metabolism. Pseudomonas aeruginosa PA14 wild type and wspF mutant that form the biofilm of distinct nature were used for the model strains to test our method. During bacterial adhesion and biofilm maturation, EIS data were collected and equivalent circuit analysis was carried out to obtain constant phase element (CPE) values representing double-layer capacitance. Since the influence by the bacterial growth-related culture media condition was eliminated by adopting fresh electrolyte at the measurement, the contribution of attached bacteria and biofilm was exclusively measured. As a result, the bacterial adhesion at the early stage of biofilm development was specifically monitored from reduction in double-layer capacitance. Particularly, the plateau in double-layer capacitance appeared upon biofilm maturation, indicating that biofilm maturation could be expected beyond this point. In conclusion, this study found that measurement of double-layer capacitance based on EIS could provide a monitoring parameter suggesting bacterial adhesion and the initiation point of biofilm maturation. PMID:21762943
Kim, Taeyoung; Kang, Junil; Lee, Joon-Hee; Yoon, Jeyong
Human plasma enhances the expression of Staphylococcal microbial surface components recognizing adhesive matrix molecules promoting biofilm formation and increases antimicrobial tolerance In Vitro
Background Microbial biofilms have been associated with the development of chronic human infections and represent a clinical challenge given their increased antimicrobial tolerance. Staphylococcus aureus is a major human pathogen causing a diverse range of diseases, of which biofilms are often involved. Staphylococcal attachment and the formation of biofilms have been shown to be facilitated by host factors that accumulate on surfaces. To better understand how host factors enhance staphylococcal biofilm formation, we evaluated the effect of whole human plasma on biofilm formation in clinical isolates of S. aureus and the expression of seven microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) known to be involved in biofilm formation by quantitative real-time PCR. We also evaluated whether plasma augmented changes in S. aureus biofilm morphology and antimicrobial resistance. Results Exposure of clinical isolates of S. aureus to human plasma (10%) within media, and to a lesser extent when coated onto plates, significantly enhanced biofilm formation in all of the clinical isolates tested. Compared to biofilms grown under non-supplemented conditions, plasma-augmented biofilms displayed significant changes in both the biofilm phenotype and cell morphology as determined by confocal scanning laser microscopy (CLSM) and scanning electron microscopy (SEM), respectively. Exposure of bacteria to plasma resulted in a significant fold-increase in MSCRAMM expression in both a time and isolate-dependent manner. Additionally, plasma-augmented biofilms displayed an increased tolerance to vancomycin compared to biofilms grown in non-supplemented media. Conclusions Collectively, these studies support previous findings demonstrating a role for host factors in biofilm formation and provide further insight into how plasma, a preferred growth medium for staphylococcal biofilm formation enhances as well as augments other intrinsic properties of S. aureus biofilms. Consequently, these findings indicate that incorporation of host factors may be necessary to better replicate in vivo conditions and for the best utility of a clinical biofilm assay to evaluate the process of biofilm formation and treatments.
Biofilm formation of Klebsiella pneumoniae on urethral catheters requires either type 1 or type 3 fimbriae
Urinary catheters are standard medical devices utilized in both hospital and nursing home settings, but are associated with a high frequency of catheter-associated urinary tract infections (CAUTI). In particular, biofilm formation on the catheter surface by uropathogens such as Klebsiella pneumoniae causes severe problems. Here we demonstrate that type 1 and type 3 fimbriae expressed by K. pneumoniae enhance biofilm formation on urinary catheters in a catheterized bladder model that mirrors the physico-chemical conditions present in catheterized patients. Furthermore, we show that both fimbrial types are able to functionally compensate for each other during biofilm formation on urinary catheters. In situ monitoring of fimbrial expression revealed that neither of the two fimbrial types is expressed when cells are grown planktonically. Interestingly, during biofilm formation on catheters, both fimbrial types are expressed, suggesting that they are both important in promoting biofilm formation on catheters. Additionally, transformed into and expressed by a nonfimbriated Escherichia coli strain, both fimbrial types significantly increased biofilm formation on catheters compared with the wild-type E. coli strain. The widespread occurrence of the two fimbrial types in different species of pathogenic bacteria stresses the need for further assessment of their role during urinary tract infections.
Stahlhut, Steen G; Struve, Carsten; Krogfelt, Karen A; Reisner, Andreas
Beta- Lactam Antibiotics Stimulate Biofilm Formation in Non-Typeable Haemophilus influenzae by Up-Regulating Carbohydrate Metabolism
Non-typeable Haemophilus influenzae (NTHi) is a common acute otitis media pathogen, with an incidence that is increased by previous antibiotic treatment. NTHi is also an emerging causative agent of other chronic infections in humans, some linked to morbidity, and all of which impose substantial treatment costs. In this study we explore the possibility that antibiotic exposure may stimulate biofilm formation by NTHi bacteria. We discovered that sub-inhibitory concentrations of beta-lactam antibiotic (i.e., amounts that partially inhibit bacterial growth) stimulated the biofilm-forming ability of NTHi strains, an effect that was strain and antibiotic dependent. When exposed to sub-inhibitory concentrations of beta-lactam antibiotics NTHi strains produced tightly packed biofilms with decreased numbers of culturable bacteria but increased biomass. The ratio of protein per unit weight of biofilm decreased as a result of antibiotic exposure. Antibiotic-stimulated biofilms had altered ultrastructure, and genes involved in glycogen production and transporter function were up regulated in response to antibiotic exposure. Down-regulated genes were linked to multiple metabolic processes but not those involved in stress response. Antibiotic-stimulated biofilm bacteria were more resistant to a lethal dose (10 �g/mL) of cefuroxime. Our results suggest that beta-lactam antibiotic exposure may act as a signaling molecule that promotes transformation into the biofilm phenotype. Loss of viable bacteria, increase in biofilm biomass and decreased protein production coupled with a concomitant up-regulation of genes involved with glycogen production might result in a biofilm of sessile, metabolically inactive bacteria sustained by stored glycogen. These biofilms may protect surviving bacteria from subsequent antibiotic challenges, and act as a reservoir of viable bacteria once antibiotic exposure has ended.
Wu, Siva; Li, Xiaojin; Gunawardana, Manjula; Maguire, Kathleen; Guerrero-Given, Debbie; Schaudinn, Christoph; Wang, Charles; Baum, Marc M.; Webster, Paul
Osteopontin Reduces Biofilm Formation in a Multi-Species Model of Dental Biofilm
Background Combating dental biofilm formation is the most effective means for the prevention of caries, one of the most widespread human diseases. Among the chemical supplements to mechanical tooth cleaning procedures, non-bactericidal adjuncts that target the mechanisms of bacterial biofilm formation have gained increasing interest in recent years. Milk proteins, such as lactoferrin, have been shown to interfere with bacterial colonization of saliva-coated surfaces. We here study the effect of bovine milk osteopontin (OPN), a highly phosphorylated whey glycoprotein, on a multispecies in vitro model of dental biofilm. While considerable research effort focuses on the interaction of OPN with mammalian cells, there are no data investigating the influence of OPN on bacterial biofilms. Methodology/Principal Findings Biofilms consisting of Streptococcus oralis, Actinomyces naeslundii, Streptococcus mitis, Streptococcus downei and Streptococcus sanguinis were grown in a flow cell system that permitted in situ microscopic analysis. Crystal violet staining showed significantly less biofilm formation in the presence of OPN, as compared to biofilms grown without OPN or biofilms grown in the presence of caseinoglycomacropeptide, another phosphorylated milk protein. Confocal microscopy revealed that OPN bound to the surface of bacterial cells and reduced mechanical stability of the biofilms without affecting cell viability. The bacterial composition of the biofilms, determined by fluorescence in situ hybridization, changed considerably in the presence of OPN. In particular, colonization of S. mitis, the best biofilm former in the model, was reduced dramatically. Conclusions/Significance OPN strongly reduces the amount of biofilm formed in a well-defined laboratory model of acidogenic dental biofilm. If a similar effect can be observed in vivo, OPN might serve as a valuable adjunct to mechanical tooth cleaning procedures.
Schlafer, Sebastian; Raarup, Merete K.; Wejse, Peter L.; Nyvad, Bente; Stadler, Brigitte M.; Sutherland, Duncan S.; Birkedal, Henrik; Meyer, Rikke L.
The control of Staphylococcus epidermidis biofilm formation and in vivo infection rates by covalently bound furanones
In order to overcome the continuing infection rate associated with biomaterials, the use of covalently bound furanones as an antibiofilm coating for biomaterials has been investigated. Furanones have previously been shown to inhibit growth of Gram-positive and Gram-negative bacteria. The aim of these studies were to covalently bind furanones to polymers and to test their efficacy for inhibiting biofilm formation
E. B. H. Hume; J. Baveja; B. Muir; T. L. Schubert; N. Kumar; S. Kjelleberg; H. J. Griesser; H. Thissen; R. Read; L. A. Poole-Warren; K. Schindhelm; M. D. P. Willcox
Variation in sessile microflora during biofilm formation on AISI304 stainless steel coupons
Coupons of stainless steel type AISI-304 were exposed to the industrial cooling system of a petrochemical plant fed by seawater from the Guanabara Bay, Rio de Janeiro, Brazil, in order to study thein situ formation of biofilms. Bacteria, microalgae and fungi were detected on the coupons as soon as 48 h after exposure. Their respective numbers were determined at times
F P de Fran�a; M T S Lutterbach
Sexual biofilm formation in Candida tropicalis opaque cells.
Candida albicans and Candida tropicalis are opportunistic fungal pathogens that can transition between white and opaque phenotypic states. White and opaque cells differ both morphologically and in their responses to environmental signals. In C. albicans, opaque cells respond to sexual pheromones by undergoing conjugation, while white cells are induced by pheromones to form sexual biofilms. Here, we show that sexual biofilm formation also occurs in C. tropicalis but, unlike C. albicans, biofilms are formed exclusively by opaque cells. C. tropicalis biofilm formation was dependent on the pheromone receptors Ste2 and Ste3, confirming the role of pheromone signalling in sexual biofilm development. Structural analysis of C. tropicalis sexual biofilms revealed stratified communities consisting of a basal layer of yeast cells and an upper layer of filamentous cells, together with an extracellular matrix. Transcriptional profiling showed that genes involved in pheromone signalling and conjugation were upregulated in sexual biofilms. Furthermore, FGR23, which encodes an agglutinin-like protein, was found to enhance both mating and sexual biofilm formation. Together, these studies reveal that C. tropicalis opaque cells form sexual biofilms with a complex architecture, and suggest a conserved role for sexual agglutinins in mediating mating, cell cohesion and biofilm formation. PMID:24612417
Jones, Stephen K; Hirakawa, Matthew P; Bennett, Richard J
Our understanding of the virulence and pathogenesis of Francisella spp. has significantly advanced in recent years, including a new understanding that this organism can form biofilms. What is known so far about Francisella spp. biofilms is summarized here and future research questions are suggested. The molecular basis of biofilm production has begun to be studied, especially the role of extracellular carbohydrates and capsule, quorum sensing and two-component signaling systems. Further work has explored the contribution of amoebae, pili, outer-membrane vesicles, chitinases, and small molecules such as c-di-GMP to Francisella spp. biofilm formation. A role for Francisella spp. biofilm in feeding mosquito larvae has been suggested. As no strong role in virulence has been found yet, Francisella spp. biofilm formation is most likely a key mechanism for environmental survival and persistence. The significance and importance of Francisella spp.�s biofilm phenotype as a critical aspect of its microbial physiology is being developed. Areas for further studies include the potential role of Francisella spp. biofilms in the infection of mammalian hosts and virulence regulation.
van Hoek, Monique L
Disinfection byproduct formation from chlorination of pure bacterial cells and pipeline biofilms.
Disinfection byproduct (DBP) formation is commonly attributed to the reaction between natural organic matters and disinfectants, yet few have considered the contribution from disinfecting bacterial materials - the essential process of water disinfection. Here, we explored the DBP formation from chlorination and chloramination of Escherichia coli and found that most selected DBPs were detectable, including trihalomethanes, haloacetonitriles, chloral hydrate, chloropicrin, and 1,1,1-trichloro-2-propanone. A positive correlation (P = 0.08-0.09) between DBP formation and the log reduction of E. coli implied that breaking down of bacterial cells released precursors for DBP formation. As Pseudomonas aeruginosa is a dominant bacterial species in pipeline biofilms, the DBP formation potentials (DBPFPs) from its planktonic cells and biofilms were characterized. Planktonic cells formed 7-11 times greater trihalomethanes per carbon of those from biofilms but significantly lower (P < 0.05) chloral hydrate, highlighting the bacterial phenotype's impact on the bacteria-derived DBPFP. Pipe material appeared to affect the DBPFP of bacteria, with 4-28% lower bromine incorporation factor for biofilms on polyvinyl chloride compared to that on galvanized zinc. This study revealed both the in situ disinfection of bacterial planktonic cells in source water and ex situ reaction between biofilms and residual chlorine in pipeline networks as hitherto unknown DBP sources in drinking water. PMID:23499193
Wang, Jun-Jian; Liu, Xin; Ng, Tsz Wai; Xiao, Jie-Wen; Chow, Alex T; Wong, Po Keung
Marine bacterial isolates inhibit biofilm formation and disrupt mature biofilms of Pseudomonas aeruginosa PAO1
According to the Centers for Disease Control and Prevention, biofilms cause 65% of infections in developed countries. Pseudomonas aeruginosa biofilm cause life threatening infections in cystic fibrosis infection and they are 1,000 times more tolerant to antibiotic\\u000a than the planktonic cells. As quorum sensing, hydrophobicity index and extracellular polysaccharide play a crucial role in\\u000a biofilm formation, extracts from 46 marine
Chari Nithya; Mansur Farzana Begum; Shunmugiah Karutha Pandian
Biodegradable polymer (PLGA) coatings featuring cinnamaldehyde and carvacrol mitigate biofilm formation.
Biofilm-associated infections are one of the leading causes of death in the United States. Although infections may be treated with antibiotics, the overuse of antibiotics has led to the spread of antibiotic resistance. Many natural antimicrobial compounds derived from edible plants are safe for human use and target bacteria nonspecifically. Therefore, they may impair biofilm formation with less evolutionary pressure on pathogens. Here, we explore the use of two natural antimicrobial compounds, cinnamaldehyde (CA, from cinnamon) and carvacrol (CARV, from oregano), for biofilm prevention. We have fabricated and characterized films that incorporate CA and CARV into the biodegradable, FDA-approved polymer poly(lactic-co-glycolic acid), PLGA. The addition of CA and CARV to PLGA films not only adds antimicrobial activity but also changes the surface properties of the films, making them more hydrophilic and therefore more resistant to bacterial attachment. An addition of 0.1% CA to a PLGA film significantly impairs biofilm development by Staphylococcus aureus, and 0.1% CARV in PLGA significantly decreases biofilm formation by both Escherichia coli and S. aureus. Pseudomonas aeruginosa, which is less susceptible to CA and CARV, was not affected by the addition of 0.1% CA or CARV to the PLGA coatings; however, P. aeruginosa biofilm was significantly reduced by 1.0% CA. These results indicate that both CA and CARV could potentially be used in low concentrations as natural additives in polymer coatings for indwelling devices to delay colonization by bacteria. PMID:22937881
Zodrow, Katherine R; Schiffman, Jessica D; Elimelech, Menachem
Non-invasive determination of conjugative transfer of plasmids bearing antibiotic-resistance genes in biofilm-bound bacteria: effects of substrate loading and antibiotic selection
Biofilms cause much of all human microbial infections. Attempts to eradicate biofilm-based infections rely on disinfectants and antibiotics. Unfortunately, biofilm bacteria are significantly less responsive to antibiotic stressors than their planktonic counterparts. Sublethal doses of antibiotics can actually enhance biofilm formation. Here, we have developed a non-invasive microscopic image analyses to quantify plasmid conjugation within a developing biofilm. Corroborating destructive samples were analyzed by a cultivation-independent flow cytometry analysis and a selective plate count method to cultivate transconjugants. Increases in substrate loading altered biofilm 3-D architecture and subsequently affected the frequency of plasmid conjugation (decreases at least two times) in the absence of any antibiotic selective pressure. More importantly, donor populations in biofilms exposed to a sublethal dose of kanamycin exhibited enhanced transfer efficiency of plasmids containing the kanamycin resistance gene, up to tenfold. However, when stressed with a different antibiotic, imipenem, transfer of plasmids containing the kanR+ gene was not enhanced. These preliminary results suggest biofilm bacteria �sense� antibiotics to which they are resistant, which enhances the spread of that resistance. Confocal scanning microscopy coupled with our non-invasive image analysis was able to estimate plasmid conjugative transfer efficiency either averaged over the entire biofilm landscape or locally with individual biofilm clusters.
Ma, Hongyan; Bryers, James D.
Non-invasive determination of conjugative transfer of plasmids bearing antibiotic-resistance genes in biofilm-bound bacteria: effects of substrate loading and antibiotic selection.
Biofilms cause much of all human microbial infections. Attempts to eradicate biofilm-based infections rely on disinfectants and antibiotics. Unfortunately, biofilm bacteria are significantly less responsive to antibiotic stressors than their planktonic counterparts. Sublethal doses of antibiotics can actually enhance biofilm formation. Here, we have developed a non-invasive microscopic image analyses to quantify plasmid conjugation within a developing biofilm. Corroborating destructive samples were analyzed by a cultivation-independent flow cytometry analysis and a selective plate count method to cultivate transconjugants. Increases in substrate loading altered biofilm 3-D architecture and subsequently affected the frequency of plasmid conjugation (decreases at least two times) in the absence of any antibiotic selective pressure. More importantly, donor populations in biofilms exposed to a sublethal dose of kanamycin exhibited enhanced transfer efficiency of plasmids containing the kanamycin resistance gene, up to tenfold. However, when stressed with a different antibiotic, imipenem, transfer of plasmids containing the kan(R+) gene was not enhanced. These preliminary results suggest biofilm bacteria "sense" antibiotics to which they are resistant, which enhances the spread of that resistance. Confocal scanning microscopy coupled with our non-invasive image analysis was able to estimate plasmid conjugative transfer efficiency either averaged over the entire biofilm landscape or locally with individual biofilm clusters. PMID:22669634
Ma, Hongyan; Bryers, James D
Epigallocatechin Gallate Inhibits Biofilm Formation by Ocular Staphylococcal Isolates
Epigallocatechin gallate (EGCg), the main polyphenol component of green tea, has several antibacterial properties. Here we show that sub-MICs of EGCg appear to decrease slime production, therefore inhibiting biofilm formation by ocular staphylococcal isolates previously characterized for the presence of ica genes by the Congo red agar plate assay and for adhesion to microtiter plates. Biofilm formation is a three-stage
Anna Rita Blanco; Andrea Sudano-Roccaro; Giovanna Carmela Spoto; Antonia Nostro; Dario Rusciano
Enterococcal surface protein, Esp, enhances biofilm formation by Enterococcus faecalis.
Enterococci play a dual role in human ecology. They serve as commensal organisms of the gastrointestinal tract and are also leading causes of multiple antibiotic-resistant hospital-acquired infection. Many nosocomial infections result from the ability of microorganisms to form biofilms. The molecular mechanisms involved in enterococcal biofilm formation are only now beginning to be understood. Enterococcal surface protein, Esp, has been reported to contribute to biofilm formation by Enterococcus faecalis. Recent studies have shown that enterococci form biofilms independently of Esp expression. To precisely determine what role Esp plays in E. faecalis biofilm formation, Esp was expressed on the cell surface of genetically well-defined, natively Esp-deficient strains, and isogenic Esp-positive and Esp-deficient strains were compared for their biofilm-forming ability. The results show that Esp expression leads to a significant increase in biofilm formation, irrespective of the strain tested. The contribution of Esp to biofilm formation was found to be most pronounced in the presence of 0.5% (wt/vol) or greater glucose. These results unambiguously define Esp as a key contributor to the ability of E. faecalis to form biofilms. PMID:15385507
Tendolkar, Preeti M; Baghdayan, Arto S; Gilmore, Michael S; Shankar, Nathan
Analysis and succession of nitrifying bacteria community structure in sequencing biofilm batch reactor.
To reveal the succession procedure of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) community structure in sequencing biofilm batch reactor (SBBR), the molecular biological techniques of denaturing gradient gel electrophoresis (DGGE), cloning, and real-time PCR were applied. DGGE showed that the structural diversity of the bacterial community increased during the biofilm formation period, and some kinds of populations had been highly preponderant consistently. The results of cloning and sequencing revealed that Nitrosomonas was the dominant species. The real-time PCR analysis indicated that the amount of the AOB increased significantly after the cultivation period, and the NOB gradually decreased. The AOB content on the 25th day was 17 times that of the 6th day. It also showed the biofilm formed successfully with accumulating nitrite and prepared to achieve the achievement of simultaneous nitrification and denitrification in SBBR. Furthermore, the ammonia-oxidizing rate was in correspondence with the NH4 (+)-N removal efficiency. PMID:24493570
Wang, Jingyin; Zhang, Chaosheng; Rong, Hongwei
Complex conductivity response to microbial growth and biofilm formation on phenanthrene spiked medium
NASA Astrophysics Data System (ADS)
Several laboratory studies have recently demonstrated the utility of geophysical methods for the investigation of microbial-induced changes over contaminated sites. However, it remains difficult to distinguish the effects due to the new physical properties imparted by microbial processes, to bacterial growth, or to the development of bacterial biofilm. We chose to study the influence of biofilm formation on geophysical response using complex conductivity measurements (0.1-1000 Hz) in phenanthrene-contaminated media. Biotic assays were conducted with two phenanthrene (PHE) degrading bacterial strains: Burkholderia sp (NAH1), which produced biofilm and Stenophomonas maltophilia (MATE10), which did not, and an abiotic control. Results showed that bacterial densities for NAH1 and MATE10 strains continuously increased at the same rate during the experiment. However, the complex conductivity signature showed noticeable differences between the two bacteria, with a phase shift of 50 mrad at 4 Hz for NAH1, which produced biofilm. Biofilm volume was quantified by Scanning Confocal Laser Microscopy (SCLM). Significant correlations were established between phase shift decrease and biofilm volume for NAH1 assays. Results suggest that complex conductivity measurements, specifically phase shift, can be a useful indicator of biofilm formation inside the overall signal of microbial activity on contaminated sites.
Albrecht, Remy; Gourry, Jean Christophe; Simonnot, Marie-Odile; Leyval, Corinne
Assessment of inhibitory effects of fluoride-coated tubes on biofilm formation by using the in vitro dental unit waterline biofilm model.
This study aimed to establish an in vitro model to simulate biofilms formed in dental unit waterlines (DUWLs) and to investigate the ability of polyvinylidene fluoride (PVDF)-coated tubes to inhibit biofilm formation using this model. The water and biofilm samples were obtained from DUWLs which had been clinically used for 2.5 years, and the predominant bacteria were identified. A conventional polyurethane tube was incubated for 24 to 96 h in the mixed flora of isolated bacteria, and the optimal incubation conditions to simulate a clinically formed biofilm were determined by observation with a scanning electron microscope. Biofilm formation on a PVDF-coated tube was observed using this in vitro model, and the adherence of different bacterial species to conventional and PVDF-coated tubes was assessed. Sphingomonas paucimobilis, Acinetobacter haemolytics, and Methylobacterium mesophilicum were predominantly isolated from contaminated DUWLs. Incubation of the polyurethane tube with the mixed flora containing these three species for 96 h resulted in the formation of a mature biofilm similar to the one clinically observed. The PVDF-coated tube was significantly less adhesive to all three bacterial species than the polyurethane tube (P < 0.05 by the Mann-Whitney U test), and the attachment of small amounts of rods was observed even after incubation with the mixed flora for 96 h. In conclusion, an in vitro biofilm model was obtained by using a mixed flora of bacteria isolated from DUWLs, and the PVDF-coated tube was found to be effective in preventing biofilm formation using this model. PMID:18676694
Yabune, Toshiaki; Imazato, Satoshi; Ebisu, Shigeyuki
Assessment of Inhibitory Effects of Fluoride-Coated Tubes on Biofilm Formation by Using the In Vitro Dental Unit Waterline Biofilm Model?
This study aimed to establish an in vitro model to simulate biofilms formed in dental unit waterlines (DUWLs) and to investigate the ability of polyvinylidene fluoride (PVDF)-coated tubes to inhibit biofilm formation using this model. The water and biofilm samples were obtained from DUWLs which had been clinically used for 2.5 years, and the predominant bacteria were identified. A conventional polyurethane tube was incubated for 24 to 96 h in the mixed flora of isolated bacteria, and the optimal incubation conditions to simulate a clinically formed biofilm were determined by observation with a scanning electron microscope. Biofilm formation on a PVDF-coated tube was observed using this in vitro model, and the adherence of different bacterial species to conventional and PVDF-coated tubes was assessed. Sphingomonas paucimobilis, Acinetobacter haemolytics, and Methylobacterium mesophilicum were predominantly isolated from contaminated DUWLs. Incubation of the polyurethane tube with the mixed flora containing these three species for 96 h resulted in the formation of a mature biofilm similar to the one clinically observed. The PVDF-coated tube was significantly less adhesive to all three bacterial species than the polyurethane tube (P < 0.05 by the Mann-Whitney U test), and the attachment of small amounts of rods was observed even after incubation with the mixed flora for 96 h. In conclusion, an in vitro biofilm model was obtained by using a mixed flora of bacteria isolated from DUWLs, and the PVDF-coated tube was found to be effective in preventing biofilm formation using this model.
Yabune, Toshiaki; Imazato, Satoshi; Ebisu, Shigeyuki
Thiophenones inhibit Staphylococcus epidermidis biofilm formation at nontoxic concentrations.
Frequent use of medical implants has led Staphylococcus epidermidis to develop into an opportunistic pathogen. The virulence is mainly linked to biofilm formation. Infections associated with biofilms are difficult to treat owing to enhanced resistance to antibiotics. Therefore, new and alternative treatments are called for. Bacterial communication is one of the regulatory mechanisms suggested to be involved in coordinating biofilm formation. In this study, we compared three communication inhibitors for preventing in vitro biofilm formation: a synthetic furanone, and two synthetic thiophenones, which are sulphur analogues of furanones. Furanones naturally source from the red macro alga Delisea pulchra. We also investigated the effect of thiophenone on transcriptional levels of genes associated with biofilm formation. We found that thiophenones were more effective in inhibiting biofilm formation than furanone, also in presence of albumin. We furthermore found that the thiophenones inhibited biofilm formation and bacterial communication more than furanones, and were less cytotoxic. The expression of the icaC and the lrgB genes, which are associated with biofilm formation, were affected by the thiophenone. PMID:22443118
L�nn-Stensrud, Jessica; Naemi, Ali-Oddin; Benneche, Tore; Petersen, Fernanda Cristina; Scheie, Anne Aamdal
Dynamics of biofilm formation during anaerobic digestion of organic waste.
Biofilm-based reactors are effectively used for wastewater treatment but are not common in biogas production. This study investigated biofilm dynamics on biofilm carriers incubated in batch biogas reactors at high and low organic loading rates for sludge from meat industry dissolved air flotation units. Biofilm formation and dynamics were studied using various microscopic techniques. Resulting micrographs were analysed for total cell numbers, thickness of biofilms, biofilm-covered surface area, and the area covered by extracellular polymeric substances (EPS). Cell numbers within biofilms (10(11)�cells�ml(-1)) were up to one order of magnitude higher compared to the numbers of cells in the fluid reactor content. Further, biofilm formation and structure mainly correlated with the numbers of microorganisms present in the fluid reactor content and the organic loading. At high organic loading (45�kg�VS�m(-3)), the thickness of the continuous biofilm layer ranged from 5 to 160�?m with an average of 51�?m and a median of 26�?m. Conversely, at lower organic loading (15�kg�VS�m(-3)), only microcolonies were detectable. Those microcolonies increased in their frequency of occurrence during ongoing fermentation. Independently from the organic loading rate, biofilms were embedded completely in EPS within seven days. The maturation and maintenance of biofilms changed during the batch fermentation due to decreasing substrate availability. Concomitant, detachment of microorganisms within biofilms was observed simultaneously with the decrease of biogas formation. This study demonstrates that biofilms of high cell densities can enhance digestion of organic waste and have positive effects on biogas production. PMID:24342346
Langer, Susanne; Schropp, Daniel; Bengelsdorf, Frank R; Othman, Maazuza; Kazda, Marian
Inhibitory effect of Ti-Ag alloy on artificial biofilm formation.
Titanium-silver (Ti-Ag) alloy has been improved for machinability and mechanical properties, but its anti-biofilm properties have not been elucidated yet. Thus, this study aimed to evaluate the effects of Ti-Ag alloy on biofilm formation and bacterial viability in comparison with pure Ti, pure Ag and silver-palladium (Ag-Pd) alloy. Biofilm formation on the metal plates was evaluated by growing Streptococcus mutans and Streptococcus sobrinus in the presence of metal plates. Bactericidal activity was evaluated using a film contact method. There were no significant differences in biofilm formation between pure Ti, pure Ag and Ag-Pd alloy, while biofilm amounts on Ti-20% Ag and Ti-25% Ag alloys were significantly lower (p<0.05). In addition, Ti-Ag alloys and pure Ti were not bactericidal, although pure Ag and Ag-Pd alloy killed bacteria. These results suggest that Ti-20% Ag and Ti-25% Ag alloys are suitable for dental material that suppresses biofilm formation without disturbing healthy oral microflora. PMID:24786344
Nakajo, Kazuko; Takahashi, Masatoshi; Kikuchi, Masafumi; Takada, Yukyo; Okuno, Osamu; Sasaki, Keiichi; Takahashi, Nobuhiro
Staphylococcus aureus autoinducer-2 quorum sensing decreases biofilm formation in an icaR-dependent manner
Background Staphylococcus aureus is an important pathogen that causes biofilm-associated infection in humans. Autoinducer 2 (AI-2), a quorum-sensing (QS) signal for interspecies communication, has a wide range of regulatory functions in both Gram-positive and Gram-negative bacteria, but its exact role in biofilm formation in S. aureus remains unclear. Results Here we demonstrate that mutation of the AI-2 synthase gene luxS in S. aureus RN6390B results in increased biofilm formation compared with the wild-type (WT) strain under static, flowing and anaerobic conditions and in a mouse model. Addition of the chemically synthesized AI-2 precursor in the luxS mutation strain (?luxS) restored the WT phenotype. Real-time RT-PCR analysis showed that AI-2 activated the transcription of icaR, a repressor of the ica operon, and subsequently a decreased level of icaA transcription, which was presumably the main reason why luxS mutation influences biofilm formation. Furthermore, we compared the roles of the agr-mediated QS system and the LuxS/AI-2 QS system in the regulation of biofilm formation using the ?luxS strain, RN6911 and the ?agr ?luxS strain. Our data indicate a cumulative effect of the two QS systems on the regulation of biofilm formation in S. aureus. Conclusion These findings demonstrate that AI-2 can decrease biofilm formation in S. aureus via an icaR-activation pathway. This study may provide clues for therapy in S. aureus biofilm-associated infection.
Role of Streptococcus gordonii Amylase-Binding Protein A in Adhesion to Hydroxyapatite, Starch Metabolism, and Biofilm Formation
Interactions between bacteria and salivary components are thought to be important in the establishment and ecology of the oral microflora. -Amylase, the predominant salivary enzyme in humans, binds to Streptococcus gordonii, a primary colonizer of the tooth. Previous studies have implicated this interaction in adhesion of the bacteria to salivary pellicles, catabolism of dietary starches, and biofilm formation. Amylase binding
Effects of Aronia melanocarpa constituents on biofilm formation of Escherichia coli and Bacillus cereus.
Many bacteria growing on surfaces form biofilms. Adaptive and genetic changes of the microorganisms in this structure make them resistant to antimicrobial agents. Biofilm-forming organisms on medical devices can pose serious threats to human health. Thus, there is a need for novel prevention and treatment strategies. This study aimed to evaluate the ability of Aronia melanocarpa extracts, subfractions and compounds to prevent biofilm formation and to inhibit bacterial growth of Escherichia coli and Bacillus cereus in vitro. It was found that several aronia substances possessed anti-biofilm activity, however, they were not toxic to the species screened. This non-toxic inhibition may confer a lower potential for resistance development compared to conventional antimicrobials. PMID:24317526
Br�unlich, Marie; �kstad, Ole A; Slimestad, Rune; Wangensteen, Helle; Malterud, Karl E; Barsett, Hilde
Antibody-Mediated Immobilization of Cryptococcus neoformans Promotes Biofilm Formation? �
Most microbes, including the fungal pathogen Cryptococcus neoformans, can grow as biofilms. Biofilms confer upon microbes a range of characteristics, including an ability to colonize materials such as shunts and catheters and increased resistance to antibiotics. Here, we provide evidence that coating surfaces with a monoclonal antibody to glucuronoxylomannan, the major component of the fungal capsular polysaccharide, immobilizes cryptococcal cells to a surface support and, subsequently, promotes biofilm formation. We used time-lapse microscopy to visualize the growth of cryptococcal biofilms, generating the first movies of fungal biofilm growth. We show that when fungal cells are immobilized using surface-attached specific antibody to the capsule, the initial stages of biofilm formation are significantly faster than those on surfaces with no antibody coating or surfaces coated with unspecific monoclonal antibody. Time-lapse microscopy revealed that biofilm growth was a dynamic process in which cells shuffled position during budding and was accompanied by emergence of planktonic variant cells that left the attached biofilm community. The planktonic variant cells exhibited mobility, presumably by Brownian motion. Our results indicate that microbial immobilization by antibody capture hastens biofilm formation and suggest that antibody coating of medical devices with immunoglobulins must exclude binding to common pathogenic microbes and the possibility that this effect could be exploited in industrial microbiology.
Robertson, Emma J.; Casadevall, Arturo
Formation of biofilms under phage predation: considerations concerning a biofilm increase.
Bacteriophages are emerging as strong candidates for combating bacterial biofilms. However, reports indicating that host populations can, in some cases, respond to phage predation by an increase in biofilm formation are of concern. This study investigates whether phage predation can enhance the formation of biofilm and if so, if this phenomenon is governed by the emergence of phage-resistance or by non-evolutionary mechanisms (eg spatial refuge). Single-species biofilms of three bacterial pathogens (Pseudomonas aeruginosa, Salmonella enterica serotype Typhimurium, and Staphylococcus aureus) were pretreated and post-treated with species-specific phages. Some of the phage treatments resulted in an increase in the levels of biofilm of their host. It is proposed that the phenotypic change brought about by acquiring phage resistance is the main reason for the increase in the level of biofilm of P. aeruginosa. For biofilms of S. aureus and S. enterica Typhimurium, although resistance was detected, increased formation of biofilm appeared to be a result of non-evolutionary mechanisms. PMID:23597188
Hosseinidoust, Zeinab; Tufenkji, Nathalie; van de Ven, Theo G M
Nucleases in Bdellovibrio bacteriovorus contribute towards efficient self-biofilm formation and eradication of preformed prey biofilms
Bdellovibrio bacteriovorus are predatory bacteria that burrow into prey bacteria and degrade their cell contents, including DNA and RNA, to grow. Their genome encodes diverse nucleases, some with potential export sequences. Transcriptomic analysis determined two candidate-predicted nuclease genes (bd1244, bd1934) upregulated upon contact with prey, which we hypothesised, may be involved in prey nucleic acid degradation. RT-PCR on total RNA from across the predatory cycle confirmed that the transcription of these genes peaks shortly after prey cell invasion, around the time that prey DNA is being degraded. We deleted bd1244 and bd1934 both singly and together and investigated their role in predation of prey cells and biofilms. Surprisingly, we found that the nuclease-mutant strains could still prey upon planktonic bacteria as efficiently as wild type and still degraded the prey genomic DNA. The Bdellovibrio nuclease mutants were less efficient at (self-) biofilm formation, and surprisingly, they showed enhanced predatory clearance of preformed prey cell biofilms relative to wild-type Bdellovibrio.
Lambert, Carey; Sockett, R Elizabeth
Nucleases in Bdellovibrio bacteriovorus contribute towards efficient self-biofilm formation and eradication of preformed prey biofilms.
Bdellovibrio bacteriovorus are predatory bacteria that burrow into prey bacteria and degrade their cell contents, including DNA and RNA, to grow. Their genome encodes diverse nucleases, some with potential export sequences. Transcriptomic analysis determined two candidate-predicted nuclease genes (bd1244, bd1934) upregulated upon contact with prey, which we hypothesised, may be involved in prey nucleic acid degradation. RT-PCR on total RNA from across the predatory cycle confirmed that the transcription of these genes peaks shortly after prey cell invasion, around the time that prey DNA is being degraded. We deleted bd1244 and bd1934 both singly and together and investigated their role in predation of prey cells and biofilms. Surprisingly, we found that the nuclease-mutant strains could still prey upon planktonic bacteria as efficiently as wild type and still degraded the prey genomic DNA. The Bdellovibrio nuclease mutants were less efficient at (self-) biofilm formation, and surprisingly, they showed enhanced predatory clearance of preformed prey cell biofilms relative to wild-type Bdellovibrio. PMID:23297829
Lambert, Carey; Sockett, R Elizabeth
Novel genes associated with biofilm formation of Actinobacillus pleuropneumoniae.
Actinobacillus pleuropneumoniae is a gram-negative bacterium and is the causative agent of swine pleuropneumonia, a highly contagious respiratory disease. Biofilm formation is an important ability possessed by numerous bacterial pathogens. The purpose of this study was to identify and characterize biofilm mutants of A. pleuropneumoniae serotype 1 strain S4074 created using a mini Tn-10 transposon. The transposon library was screened to identify mutants with a modified ability to form biofilms in polystyrene microtiter plates. A total of 1200 mutants were screened and the analysis identified 24 mutants that exhibited abnormal biofilm formation, at least 16 unique genes were identified. Most genes identified in the enhanced-biofilm mutants encoded proteins with unknown functions, whereas most genes identified in the biofilm-reduced mutants encoded proteins related to transport, protein synthesis and nucleic acid synthesis. Approximately 50% of genes, including hns, potD2, ptsI, tig and rpmF, identified in our screen have been previously associated with biofilm formation in A. pleuropneumoniae and other bacterial species, and thus validated the screening method. The rest of genes identified, such as APL_0049, APL_0637 and APL_1572, have not been previously associated with biofilm formation. Interestingly, gene APL_0049 was previously seen among the genes differentially expressed during a natural infection of pig lungs. Preliminary characterization of the mutants was also initiated by assessing their hydrophobicity, their biofilm matrix composition and their ability to adhere to a polystyrene surface or NPTr cells. Based on the preliminary characterization, some of the mutants identified appear to have deficiencies during the initial attachment or growth of the biofilm. In conclusion, transposon mutagenesis analysis allowed the identification of new genes associated with biofilm formation in A. pleuropneumoniae. PMID:21530112
Grasteau, Alexandra; Tremblay, Yannick D N; Labrie, Jos�e; Jacques, Mario
Contribution of Cell Elongation to the Biofilm Formation of Pseudomonas aeruginosa during Anaerobic Respiration
Pseudomonas aeruginosa, a gram-negative bacterium of clinical importance, forms more robust biofilm during anaerobic respiration, a mode of growth presumed to occur in abnormally thickened mucus layer lining the cystic fibrosis (CF) patient airway. However, molecular basis behind this anaerobiosis-triggered robust biofilm formation is not clearly defined yet. Here, we identified a morphological change naturally accompanied by anaerobic respiration in P. aeruginosa and investigated its effect on the biofilm formation in vitro. A standard laboratory strain, PAO1 was highly elongated during anaerobic respiration compared with bacteria grown aerobically. Microscopic analysis demonstrated that cell elongation likely occurred as a consequence of defective cell division. Cell elongation was dependent on the presence of nitrite reductase (NIR) that reduces nitrite (NO2?) to nitric oxide (NO) and was repressed in PAO1 in the presence of carboxy-PTIO, a NO antagonist, demonstrating that cell elongation involves a process to respond to NO, a spontaneous byproduct of the anaerobic respiration. Importantly, the non-elongated NIR-deficient mutant failed to form biofilm, while a mutant of nitrate reductase (NAR) and wild type PAO1, both of which were highly elongated, formed robust biofilm. Taken together, our data reveal a role of previously undescribed cell biological event in P. aeruginosa biofilm formation and suggest NIR as a key player involved in such process.
Park, Yongjin; Yoon, Sang Sun
Effect of Cinnamon Oil on icaA Expression and Biofilm Formation by Staphylococcus epidermidis?
Staphylococcus epidermidis is notorious for its biofilm formation on medical devices, and novel approaches to prevent and kill S. epidermidis biofilms are desired. In this study, the effect of cinnamon oil on planktonic and biofilm cultures of clinical S. epidermidis isolates was evaluated. Initially, susceptibility to cinnamon oil in planktonic cultures was compared to the commonly used antimicrobial agents chlorhexidine, triclosan, and gentamicin. The MIC of cinnamon oil, defined as the lowest concentration able to inhibit visible microbial growth, and the minimal bactericidal concentration, the lowest concentration required to kill 99.9% of the bacteria, were determined using the broth microdilution method and plating on agar. A checkerboard assay was used to evaluate the possible synergy between cinnamon oil and the other antimicrobial agents. The effect of cinnamon oil on biofilm growth was studied in 96-well plates and with confocal laser-scanning microscopy (CLSM). Biofilm susceptibility was determined using a metabolic 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Real-time PCR analysis was performed to determine the effect of sub-MIC concentrations of cinnamon oil on expression of the biofilm-related gene, icaA. Cinnamon oil showed antimicrobial activity against both planktonic and biofilm cultures of clinical S. epidermidis strains. There was only a small difference between planktonic and biofilm MICs, ranging from 0.5 to 1% and 1 to 2%, respectively. CLSM images indicated that cinnamon oil is able to detach and kill existing biofilms. Thus, cinnamon oil is an effective antimicrobial agent to combat S. epidermidis biofilms.
Nuryastuti, Titik; van der Mei, Henny C.; Busscher, Henk J.; Iravati, Susi; Aman, Abu T.; Krom, Bastiaan P.
Analysis of the mucR gene regulating biosynthesis of exopolysaccharides: implications for biofilm formation in Sinorhizobium meliloti Rm1021.
Bacterial surface polysaccharides are crucial for establishment of successful rhizobia-legume symbiosis, and in most bacteria, are also critical for biofilm formation and surface colonization. In Sinorhizobium meliloti, the regulatory protein MucR controls exopolysaccharide production. To clarify the relationship between exopolysaccharide synthesis and biofilm formation, we studied mucR expression under growth conditions that influence attachment to polyvinylchloride, developed a microtiter plate assay to quantify biofilm formation in S. meliloti strain Rm1021 and mutants defective in succinoglycan (EPS I) and/or galactoglucan (EPS II) production, and analyzed expression of EPS I and EPS II genes by quantitative reverse transcriptase-PCR. Consistent with previous studies of planktonic bacteria, we found that disruption of the mucR gene in Rm1021 biofilms increased EPS II, but reduced EPS I gene expression. mucR expression was not affected by environmental conditions that influence biofilm formation on polyvinylchloride, and biofilm formation by Rm1021 was independent of exopolysaccharide synthesis. Other factors on the Rm1021 cell surface, and growth conditions, presumably regulate attachment and/or growth as a biofilm on polyvinylchloride. PMID:19929968
Rinaudi, Luciana V; Sorroche, Fernando; Zorreguieta, Angeles; Giordano, Walter
[Methods for detection of biofilm formation in routine microbiological practice].
The increasing use of catheters, artificial implants and antimicrobials as well as high numbers of immunocompromised patients are major causes for concern over biofilm infections. These infections are characterized particularly by high resistance to antimicrobials and formation of persistent foci that may complicate therapy. Therefore, detection of biofilm formation is of high relevance to the clinician and his/her approach to the treatment. Reliable and sensitive methods for detection of this pathogenicity factor in clinically important organisms, suitable for use in routine microbiological laboratories, are needed for this purpose. Currently, a wide array of techniques are available for detection of this virulence factor, such as biofilm visualization by microscopy, culture detection, detection of particular components, detection of physical and chemical differences between biofilm-positive organisms and their planktonic forms and detection of genes responsible for biofilm formation. Since each of these methods has limitations, the best results can be achieved by combining different approaches. PMID:16528896
R?zicka, F; Hol�, V; Votava, M
Micropatterned biofilm formations by laminar flow-templating.
We present a microfluidic device capable of patterning linear biofilm formations using a flow templating approach. We describe the design considerations and fabrication methodology of a two level flow-templating micro-bioreactor (FT-?BR), which generates a biofilm growth stream surrounded on 3 sides by a growth inhibiting confinement stream. Through a combination of experiments and simulations we comprehensively evaluate and exploit control parameters to manipulate the biofilm growth template stream dimensions. The FT-?BR is then used to grow biofilm patterns with controllable dimensions. A proof-of-principle study using the device demonstrates its utility in conducting biofilm growth rate measurements under different shear stress environments. This opens the way for quantitative studies into the effects of the local shear environment on biofilm properties and for the synthesis of a new generation of functional biomaterials with controllable properties. PMID:24722812
Aznaveh, Nahid Babaei; Safdar, Muhammad; Wolfaardt, Gideon; Greener, Jesse
Autoinducer 2 of Fusobacterium nucleatum as a target molecule to inhibit biofilm formation of periodontopathogens.
Periodontitis is initiated by bacteria in subgingival biofilms, which are composed mostly of Gram-negative anaerobes. Autoinducer 2 (AI-2) is a universal quorum sensing (QS) molecule that mediates intergeneric signalling in multispecies bacterial communities and may induce biofilm formation. As Fusobacterium nucleatum is the major coaggregation bridge organism that links early colonising commensals and late pathogenic colonisers in dental biofilms via the accretion of periodontopathogens, we hypothesised that AI-2 of F. nucleatum contributes to this interspecies interaction, and interruption of this signalling could result in the inhibition of biofilm formation of periodontopathogens. To test this hypothesis, we evaluated the effect of partially purified F. nucleatum AI-2 on monospecies biofilm as well as mutualistic interactions between F. nucleatum and the so-called 'red complex' (Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia). Then we tested the effect of two QS inhibitors (QSIs), (5Z)-4-bromo-5-(bromomethylene)-2(5H)-furanone (furanone compound) and d-ribose, on AI-2-induced biofilm formation and coaggregation. F. nucleatum AI-2 remarkably induced biofilm growth of single and dual species and coaggregation between F. nucleatum and each species of the 'red complex', all of which were inhibited by the QSIs. F. nucleatum AI-2 induced the expression of the representative adhesion molecules of the periodontopathogens, which were inhibited by the QSIs. Our results demonstrate that F. nucleatum AI-2 plays an important role in inter- and intraspecies interactions between periodontopathogens via enhanced expression of adhesion molecules and may be a target for the inhibition of pathogenic dental biofilm formation. PMID:22633049
Jang, Yun-Ji; Choi, Yu-Jung; Lee, Sung-Hoon; Jun, Hye-Kyoung; Choi, Bong-Kyu
Novel genes associated with biofilm formation of Actinobacillus pleuropneumoniae
Actinobacillus pleuropneumoniae is a Gram-negative bacterium and is the causative agent of swine pleuropneumonia, a highly contagious respiratory disease. Biofilm formation is an important ability possessed by numerous bacterial pathogens. The purpose of this study was to identify and characterize biofilm mutants of A. pleuropneumoniae serotype 1 strain S4074 created using a mini Tn-10 transposon. The transposon library was screened
Alexandra Grasteau; Yannick D. N. Tremblay; Jos�e Labrie; Mario Jacques
Biofilm formation on nanostructured hydroxyapatite-coated titanium.
Biofilm formation on medical devices is a common cause of implant failure, especially regarding implants that breach the epithelial tissue, so-called transcutaneous implants. Nanotechnology and the development of new nanomaterials have given the opportunity to design nanotextured implant surfaces. Such surfaces have been studied using various in vitro methods showing that nanosized features strongly benefit bone cell growth. However, little is known on how nanostructured features affect biofilm formation. The aim of this study was therefore to examine the shape- and chemical-dependent effect of a nanostructured hydroxyapatite (HA) coating on the degree of Staphylococcus epidermidis biofilm formation. Three different types of nanosized HA particles having different shapes and calcium to phosphate ratios were compared to uncoated turned titanium using safranin stain in a biofilm assay and confocal laser scanning microscopy (CLSM) for assessment of biofilm biomass and bacterial volume, respectively. No difference in biofilm biomass was detected for the various surfaces after 6 h incubation with S. epidermidis. Additionally, image analysis of CLSM Z-stacks confirmed the biofilm assay and showed similar results. In conclusion, the difference in nanomorphology and chemical composition of the surface coatings did not influence the adhesion and biofilm formation of S. epidermidis. PMID:23589449
Westas, Emma; Gillstedt, Martin; L�nn-Stensrud, Jessica; Bruzell, Ellen; Andersson, Martin
Vaccination with SesC Decreases Staphylococcus epidermidis Biofilm Formation
The increased use of medical implants has resulted in a concomitant rise in device-related infections. The majority of these infections are caused by Staphylococcus epidermidis biofilms. Immunoprophylaxis and immunotherapy targeting in vivo-expressed, biofilm-associated, bacterial cell surface-exposed proteins are promising new approaches to prevent and treat biofilm-related infections, respectively. Using an in silico procedure, we identified 64 proteins that are predicted to be S. epidermidis surface exposed (Ses), of which 36 were annotated as (conserved) hypothetical. Of these 36 proteins, 5 proteins�3 LPXTG motif-containing proteins (SesL, SesB, and SesC) and 2 of the largest ABC transporters (SesK and SesM)�were selected for evaluation as vaccine candidates. This choice was based on protein size, number of antigenic determinants, or the established role in S. epidermidis biofilm formation of the protein family to which the candidate protein belongs. Anti-SesC antibodies exhibited the greatest inhibitory effect on S. epidermidis biofilm formation in vitro and on colonization and infection in a mouse jugular vein catheter infection model that includes biofilms and organ infections. Active vaccination with a recombinant truncated SesC inhibited S. epidermidis biofilm formation in a rat model of subcutaneous foreign body infection. Antibodies to SesC were shown to be opsonic by an in vitro opsonophagocytosis assay. We conclude that SesC is a promising target for antibody mediated strategies against S. epidermidis biofilm formation.
Shahrooei, Mohammad; Hira, Vishal; Khodaparast, Laleh; Khodaparast, Ladan; Stijlemans, Benoit; Kucharikova, Sona; Burghout, Peter; Hermans, Peter W. M.
Enterococcal Surface Protein, Esp, Enhances Biofilm Formation by Enterococcus faecalis
Enterococci play a dual role in human ecology. They serve as commensal organisms of the gastrointestinal tract and are also leading causes of multiple antibiotic-resistant hospital-acquired infection. Many nosocomial infections result from the ability of microorganisms to form biofilms. The molecular mechanisms involved in enterococcal biofilm formation are only now beginning to be understood. Enterococcal surface protein, Esp, has been
Preeti M. Tendolkar; Arto S. Baghdayan; Michael S. Gilmore; Nathan Shankar
Effect of sub-minimum inhibitory concentrations of ciprofloxacin, amikacin and colistin on biofilm formation and virulence factors of Escherichia coli planktonic and biofilm forms isolated from human urine
The aim of this study was to determine the effect of subinhibitory concentrations (sub-MICs) of ciprofloxacin, amikacin and colistin on biofilm formation, motility, curli fimbriae formation by planktonic and biofilm cells of E. coli strains isolated from the urine of patients with various urinary system infections. Quantification of biofilm formation was carried out using a microtiter plate assay and a spectrophotometric method. Bacterial enumeration was used to assess the viability of bacteria in the biofilm. Curli expression was determined by using YESCA agar supplemented with congo red. Using motility agar the ability to move was examined. All the antibiotics used at sub-MICs reduced biofilm formation in vitro, decreased the survival of bacteria, but had no effect on the motility of planktonic as well as biofilm cells. The inhibitory effect of sub-MICs of antimicrobial agents on curli fimbriae formation was dependent on the form in which the bacteria occurred, incubation time and antibiotic used. Our results clearly show that all the three antibiotics tested reduce biofilm production, interfere with curli expression but do not influence motility. This study suggests that ciprofloxacin, amikacin and colistin may be useful in the treatment of biofilm-associated infections caused by E. coli strains.
Wojnicz, Dorota; Tichaczek-Goska, Dorota
Biofilm formation as a function of adhesin, growth medium, substratum and strain type
Biofilm formation is involved in the majority of bacterial infections. Comparing six Escherichia coli and Klebsiella pneumoniae isolates revealed significant differences in biofilm formation depending on the growth medium. Fimbriae are known to be involved in biofilm formation, and type 1, F1C and P fimbriae were seen to influence biofilm formation significantly different depending on strain background, growth media and
Viktoria Hancock; Ingun Lund Wits�; Per Klemm
OmpA-Mediated Biofilm Formation Is Essential for the Commensal Bacterium Sodalis glossinidius To Colonize the Tsetse Fly Gut
Many bacteria successfully colonize animals by forming protective biofilms. Molecular processes that underlie the formation and function of biofilms in pathogenic bacteria are well characterized. In contrast, the relationship between biofilms and host colonization by symbiotic bacteria is less well understood. Tsetse flies (Glossina spp.) house 3 maternally transmitted symbionts, one of which is a commensal (Sodalis glossinidius) found in several host tissues, including the gut. We determined that Sodalis forms biofilms in the tsetse gut and that this process is influenced by the Sodalis outer membrane protein A (OmpA). Mutant Sodalis strains that do not produce OmpA (Sodalis ?OmpA mutants) fail to form biofilms in vitro and are unable to colonize the tsetse gut unless endogenous symbiotic bacteria are present. Our data indicate that in the absence of biofilms, Sodalis ?OmpA mutant cells are exposed to and eliminated by tsetse's innate immune system, suggesting that biofilms help Sodalis evade the host immune system. Tsetse is the sole vector of pathogenic African trypanosomes, which also reside in the fly gut. Acquiring a better understanding of the dynamics that promote Sodalis colonization of the tsetse gut may enhance the development of novel disease control strategies.
Maltz, Michele A.; Weiss, Brian L.; O'Neill, Michelle; Wu, Yineng
Small RNAs regulating biofilm formation and outer membrane homeostasis.
Nowadays, the identification of small non-coding RNAs takes a prominent role in deciphering complex bacterial phenotypes. Evidences are given that the post-transcriptional layer of regulation mediated by sRNAs plays an important role in the formation of bacterial biofilms. These sRNAs exert their activity on various targets, be it directly or indirectly linked to biofilm formation. First, and best described, are the sRNAs that act in core regulatory pathways of biofilm formation, such as those regulating motility and matrix production. Second, overlaps between the regulation of biofilm formation and the outer membrane (OM) are becoming obvious. Additionally, different studies indicate that defects in the OM itself affect biofilm formation through this shared cascade, thereby forming a feedback mechanism. Interestingly, it is known that the OM itself is extensively regulated by different sRNAs. Third, biofilms are also linked to global metabolic changes. There is also evidence that metabolic pathways and the process of biofilm formation share sRNAs. PMID:23324602
Van Puyvelde, Sandra; Steenackers, Hans P; Vanderleyden, Jos
Involvement of Iron in Biofilm Formation by Staphylococcus aureus
Staphylococcus aureus is a human pathogen that forms biofilm on catheters and medical implants. The authors' earlier study established that 1,2,3,4,6-penta-O-galloyl-?-D-glucopyranose (PGG) inhibits biofilm formation by S. aureus by preventing the initial attachment of the cells to a solid surface and reducing the production of polysaccharide intercellular adhesin (PIA). Our cDNA microarray and MALDI-TOF mass spectrometric studies demonstrate that PGG treatment causes the expression of genes and proteins that are normally expressed under iron-limiting conditions. A chemical assay using ferrozine verifies that PGG is a strong iron chelator that depletes iron from the culture medium. This study finds that adding FeSO4 to a medium that contains PGG restores the biofilm formation and the production of PIA by S. aureus SA113. The requirement of iron for biofilm formation by S. aureus SA113 can also be verified using a semi-defined medium, BM, that contains an iron chelating agent, 2, 2?-dipyridyl (2-DP). Similar to the effect of PGG, the addition of 2-DP to BM medium inhibits biofilm formation and adding FeSO4 to BM medium that contains 2-DP restores biofilm formation. This study reveals an important mechanism of biofilm formation by S. aureus SA113.
Huang, Hsiu-Yun; Cheng, Yi-Ching
Expression of Xylella fastidiosa Fimbrial and Afimbrial Proteins during Biofilm Formation?
Complete sequencing of the Xylella fastidiosa genome revealed characteristics that have not been described previously for a phytopathogen. One characteristic of this genome was the abundance of genes encoding proteins with adhesion functions related to biofilm formation, an essential step for colonization of a plant host or an insect vector. We examined four of the proteins belonging to this class encoded by genes in the genome of X. fastidiosa: the PilA2 and PilC fimbrial proteins, which are components of the type IV pili, and XadA1 and XadA2, which are afimbrial adhesins. Polyclonal antibodies were raised against these four proteins, and their behavior during biofilm development was assessed by Western blotting and immunofluorescence assays. In addition, immunogold electron microscopy was used to detect these proteins in bacteria present in xylem vessels of three different hosts (citrus, periwinkle, and hibiscus). We verified that these proteins are present in X. fastidiosa biofilms but have differential regulation since the amounts varied temporally during biofilm formation, as well as spatially within the biofilms. The proteins were also detected in bacteria colonizing the xylem vessels of infected plants.
Caserta, R.; Takita, M. A.; Targon, M. L.; Rosselli-Murai, L. K.; de Souza, A. P.; Peroni, L.; Stach-Machado, D. R.; Andrade, A.; Labate, C. A.; Kitajima, E. W.; Machado, M. A.; de Souza, A. A.
Expression of Xylella fastidiosa fimbrial and afimbrial proteins during biofilm formation.
Complete sequencing of the Xylella fastidiosa genome revealed characteristics that have not been described previously for a phytopathogen. One characteristic of this genome was the abundance of genes encoding proteins with adhesion functions related to biofilm formation, an essential step for colonization of a plant host or an insect vector. We examined four of the proteins belonging to this class encoded by genes in the genome of X. fastidiosa: the PilA2 and PilC fimbrial proteins, which are components of the type IV pili, and XadA1 and XadA2, which are afimbrial adhesins. Polyclonal antibodies were raised against these four proteins, and their behavior during biofilm development was assessed by Western blotting and immunofluorescence assays. In addition, immunogold electron microscopy was used to detect these proteins in bacteria present in xylem vessels of three different hosts (citrus, periwinkle, and hibiscus). We verified that these proteins are present in X. fastidiosa biofilms but have differential regulation since the amounts varied temporally during biofilm formation, as well as spatially within the biofilms. The proteins were also detected in bacteria colonizing the xylem vessels of infected plants. PMID:20472735
Caserta, R; Takita, M A; Targon, M L; Rosselli-Murai, L K; de Souza, A P; Peroni, L; Stach-Machado, D R; Andrade, A; Labate, C A; Kitajima, E W; Machado, M A; de Souza, A A
Occurrence of nitrifiers and diversity of ammonia-oxidizing bacteria in developing drinking water biofilms
We studied the population dynamics of nitrifying bacteria during the development of biofilms up to 233 or 280 days on polyvinylchloride pipes connected to two full-scale drinking water distribution networks supplying processed and chloraminated surface water. The numbers of nitrifiers in biofilms were enumerated at intervals of 10�64 days by the most probable number (MPN) method at waterworks and at
Mari T. T. Lipponen; Pertti J. Martikainen; Ritva E. Vasara; Kristina Servomaa; Outi Zacheus; Merja H. Kontro
Initial Phases of Biofilm Formation in Shewanella oneidensis MR-1
Shewanella oneidensis MR-1 is a facultative Fe(III)- and Mn(IV)-reducing microorganism and serves as a model for studying microbially induced dissolution of Fe or Mn oxide minerals as well as biogeochemical cycles. In soil and sediment environments, S. oneidensis biofilms form on mineral surfaces and are critical for mediating the metabolic interaction between this microbe and insoluble metal oxide phases. In order to develop an understanding of the molecular basis of biofilm formation, we investigated S. oneidensis biofilms developing on glass surfaces in a hydrodynamic flow chamber system. After initial attachment, growth of microcolonies and lateral spreading of biofilm cells on the surface occurred simultaneously within the first 24 h. Once surface coverage was almost complete, biofilm development proceeded with extensive vertical growth, resulting in formation of towering structures giving rise to pronounced three-dimensional architecture. Biofilm development was found to be dependent on the nutrient conditions, suggesting a metabolic control. In global transposon mutagenesis, 173 insertion mutants out of 15,000 mutants screened were identified carrying defects in initial attachment and/or early stages in biofilm formation. Seventy-one of those mutants exhibited a nonswimming phenotype, suggesting a role of swimming motility or motility elements in biofilm formation. Disruption mutations in motility genes (flhB, fliK, and pomA), however, did not affect initial attachment but affected progression of biofilm development into pronounced three-dimensional architecture. In contrast, mutants defective in mannose-sensitive hemagglutinin type IV pilus biosynthesis and in pilus retraction (pilT) showed severe defects in adhesion to abiotic surfaces and biofilm formation, respectively. The results provide a basis for understanding microbe-mineral interactions in natural environments.
Thormann, Kai M.; Saville, Renee M.; Shukla, Soni; Pelletier, Dale A.; Spormann, Alfred M.
Biofilm formation of mucosa-associated methanoarchaeal strains
Although in nature most microorganisms are known to occur predominantly in consortia or biofilms, data on archaeal biofilm formation are in general scarce. Here, the ability of three methanoarchaeal strains, Methanobrevibacter smithii and Methanosphaera stadtmanae, which form part of the human gut microbiota, and the Methanosarcina mazei strain G�1 to grow on different surfaces and form biofilms was investigated. All three strains adhered to the substrate mica and grew predominantly as bilayers on its surface as demonstrated by confocal laser scanning microscopy analyses, though the formation of multi-layered biofilms of Methanosphaera stadtmanae and Methanobrevibacter smithii was observed as well. Stable biofilm formation was further confirmed by scanning electron microscopy analysis. Methanosarcina mazei and Methanobrevibacter smithii also formed multi-layered biofilms in uncoated plastic ?-dishesTM, which were very similar in morphology and reached a height of up to 40 ?m. In contrast, biofilms formed by Methanosphaera stadtmanae reached only a height of 2 ?m. Staining with the two lectins ConA and IB4 indicated that all three strains produced relatively low amounts of extracellular polysaccharides most likely containing glucose, mannose, and galactose. Taken together, this study provides the first evidence that methanoarchaea can develop and form biofilms on different substrates and thus, will contribute to our knowledge on the appearance and physiological role of Methanobrevibacter smithii and Methanosphaera stadtmanae in the human intestine.
Bang, Corinna; Ehlers, Claudia; Orell, Alvaro; Prasse, Daniela; Spinner, Marlene; Gorb, Stanislav N.; Albers, Sonja-Verena; Schmitz, Ruth A.
Biofilm formation by zygomycetes: quantification, structure and matrix composition.
Most studies on fungal biofilms have focused on Candida in yeasts and Aspergillus in mycelial fungi. To the authors' knowledge, biofilm formation by zygomycetes has not been reported previously. In this study, the biofilm-forming capacity of Rhizopus oryzae, Lichtheimia corymbifera, Rhizomucor pusillus and Apophysomyces elegans was evaluated. At appropriate seeding spore densities, Rhp. oryzae (10? c.f.u. ml?�, L. corymbifera (10? c.f.u. ml?�) and Rhm. pusillus (10? c.f.u. ml?�) produced highly intertwined, adherent structures on flat-bottomed polystyrene microtitre plates after 24 h at 37 �C. The adhered fungal hyphae were encased in an extracellular matrix, as confirmed by phase-contrast and confocal microscopy. The thickness of Rhp. oryzae, L. corymbifera and Rhm. pusillus biofilms was 109.67�10.02, 242�23.07 and 197�9.0 �m (mean�sd), respectively. Biochemical characterization of the biofilm matrix indicated the presence of glucosamine, constituting 74.54-82.22?% of its dry weight, N-acetylglucosamine, glucose and proteins. Adherence and biofilm formation were not observed in A. elegans. Although A. elegans spores germinated at all three seeding densities tested (1�10?, 1�10? and 1�10? c.f.u. ml?�), no significant difference was observed (P>0.05) between the A??? of wells inoculated with A. elegans and the cut-off A??? for biofilm detection. This study highlights the potential for biofilm formation by at least three medically important species of zygomycetes. PMID:21636650
Singh, Rachna; Shivaprakash, M R; Chakrabarti, Arunaloke
Virstatin inhibits biofilm formation and motility of Acinetobacter baumannii
Background Acinetobacter baumannii has emerged as an opportunistic nosocomial pathogen causing infections worldwide. One reason for this emergence is due to its natural ability to survive in the hospital environment, which may be explained by its capacity to form biofilms. Cell surface appendages are important determinants of the A. baumannii biofilm formation and as such constitute interesting targets to prevent the development of biofilm-related infections. A chemical agent called virstatin was recently described to impair the virulence of Vibrio cholerae by preventing the expression of its virulence factor, the toxin coregulated pilus (type IV pilus). The objective of this work was to investigate the potential effect of virstatin on A. baumannii biofilms. Results After a dose�response experiment, we determined that 100�?M virstatin led to an important decrease (38%) of biofilms formed by A. baumannii ATCC17978 grown under static mode. We demonstrated that the production of biofilms grown under dynamic mode was also delayed and reduced. The biofilm susceptibility to virstatin was then tested for 40 clinical and reference A. baumannii strains. 70% of the strains were susceptible to virstatin (with a decrease of 10 to 65%) when biofilms grew in static mode, whereas 60% of strains respond to the treatment when their biofilms grew in dynamic mode. As expected, motility and atomic force microscopy experiments showed that virstatin acts on the A. baumannii pili biogenesis. Conclusions By its action on pili biogenesis, virstatin demonstrated a very promising antibiofilm activity affecting more than 70% of the A. baumannii clinical isolates.
Biofilm Formation by Campylobacter jejuni Is Increased under Aerobic Conditions ?
The microaerophilic human pathogen Campylobacter jejuni is the leading cause of food-borne bacterial gastroenteritis in the developed world. During transmission through the food chain and the environment, the organism must survive stressful environmental conditions, particularly high oxygen levels. Biofilm formation has been suggested to play a role in the environmental survival of this organism. In this work we show that C. jejuni NCTC 11168 biofilms developed more rapidly under environmental and food-chain-relevant aerobic conditions (20% O2) than under microaerobic conditions (5% O2, 10% CO2), although final levels of biofilms were comparable after 3 days. Staining of biofilms with Congo red gave results similar to those obtained with the commonly used crystal violet staining. The level of biofilm formation by nonmotile aflagellate strains was lower than that observed for the motile flagellated strain but nonetheless increased under aerobic conditions, suggesting the presence of flagellum-dependent and flagellum-independent mechanisms of biofilm formation in C. jejuni. Moreover, preformed biofilms shed high numbers of viable C. jejuni cells into the culture supernatant independently of the oxygen concentration, suggesting a continuous passive release of cells into the medium rather than a condition-specific active mechanism of dispersal. We conclude that under aerobic or stressful conditions, C. jejuni adapts to a biofilm lifestyle, allowing survival under detrimental conditions, and that such a biofilm can function as a reservoir of viable planktonic cells. The increased level of biofilm formation under aerobic conditions is likely to be an adaptation contributing to the zoonotic lifestyle of C. jejuni.
Reuter, Mark; Mallett, Arthur; Pearson, Bruce M.; van Vliet, Arnoud H. M.
Biofilm Reactor for a Bacteria Fuel Cell System.
National Technical Information Service (NTIS)
A biofilm reactor has been investigated to replace a traditional bioreactor for a bacterial fuel cell system. In the fuel cell system studied, the biofilm reactor is used to separate biological reaction from electrode (anode) reaction. The reactor is made...
X. Zhang A. Halme
Influence of Calcium in Extracellular DNA Mediated Bacterial Aggregation and Biofilm Formation
Calcium (Ca2+) has an important structural role in guaranteeing the integrity of the outer lipopolysaccharide layer and cell walls of bacterial cells. Extracellular DNA (eDNA) being part of the slimy matrix produced by bacteria promotes biofilm formation through enhanced structural integrity of the matrix. Here, the concurrent role of Ca2+ and eDNA in mediating bacterial aggregation and biofilm formation was studied for the first time using a variety of bacterial strains and the thermodynamics of DNA to Ca2+ binding. It was found that the eDNA concentrations under both planktonic and biofilm growth conditions were different among bacterial strains. Whilst Ca2+ had no influence on eDNA release, presence of eDNA by itself favours bacterial aggregation via attractive acid-base interactions in addition, its binding with Ca2+ at biologically relevant concentrations was shown further increase in bacterial aggregation via cationic bridging. Negative Gibbs free energy (?G) values in iTC data confirmed that the interaction between DNA and Ca2+ is thermodynamically favourable and that the binding process is spontaneous and exothermic owing to its highly negative enthalpy. Removal of eDNA through DNase I treatment revealed that Ca2+ alone did not enhance cell aggregation and biofilm formation. This discovery signifies the importance of eDNA and concludes that existence of eDNA on bacterial cell surfaces is a key facilitator in binding of Ca2+ to eDNA thereby mediating bacterial aggregation and biofilm formation.
Koop, Leena; Wong, Yie Kuan; Ahmed, Safia; Siddiqui, Khawar Sohail; Manefield, Mike
Unsaturated Fatty Acid, cis-2-Decenoic Acid, in Combination with Disinfectants or Antibiotics Removes Pre-Established Biofilms Formed by Food-Related Bacteria.
Biofilm formation by food-related bacteria and food-related pathogenesis are significant problems in the food industry. Even though much disinfection and mechanical procedure exist for removal of biofilms, they may fail to eliminate pre-established biofilms. cis-2 decenoic acid (CDA), an unsaturated fatty acid messenger produced by Pseudomonas aeruginosa, is reportedly capable of inducing the dispersion of established biofilms by multiple types of microorganisms. However, whether CDA has potential to boost the actions of certain antimicrobials is unknown. Here, the activity of CDA as an inducer of pre-established biofilms dispersal, formed by four main food pathogens; Staphylococcus aureus, Bacillus cereus, Salmonella enterica and E. coli, was measured using both semi-batch and continuous cultures bioassays. To assess the ability of CDA combined biocides treatments to remove pre-established biofilms formed on stainless steel discs, CFU counts were performed for both treated and untreated cultures. Eradication of the biofilms by CDA combined antibiotics was evaluated using crystal violet staining. The effect of CDA combined treatments (antibiotics and disinfectants) on biofilm surface area and bacteria viability was evaluated using fluorescence microscopy, digital image analysis and LIVE/DEAD staining. MICs were also determined to assess the probable inhibitory effects of CDA combined treatments on the growth of tested microorganisms' planktonic cells. Treatment of pre-established biofilms with only 310 nM CDA resulted in at least two-fold increase in the number of planktonic cells in all cultures. While antibiotics or disinfectants alone exerted a trivial effect on CFU counts and percentage of surface area covered by the biofilms, combinational treatments with both 310 nM CDA and antibiotics or disinfectants led to approximate 80% reduction in biofilm biomass. These data suggests that combined treatments with CDA would pave the way toward developing new strategies to control biofilms with widespread applications in industry as well as medicine. PMID:25000301
Sepehr, Shayesteh; Rahmani-Badi, Azadeh; Babaie-Naiej, Hamta; Soudi, Mohammad Reza
Unsaturated Fatty Acid, cis-2-Decenoic Acid, in Combination with Disinfectants or Antibiotics Removes Pre-Established Biofilms Formed by Food-Related Bacteria
Biofilm formation by food-related bacteria and food-related pathogenesis are significant problems in the food industry. Even though much disinfection and mechanical procedure exist for removal of biofilms, they may fail to eliminate pre-established biofilms. cis-2 decenoic acid (CDA), an unsaturated fatty acid messenger produced by Pseudomonas aeruginosa, is reportedly capable of inducing the dispersion of established biofilms by multiple types of microorganisms. However, whether CDA has potential to boost the actions of certain antimicrobials is unknown. Here, the activity of CDA as an inducer of pre-established biofilms dispersal, formed by four main food pathogens; Staphylococcus aureus, Bacillus cereus, Salmonella enterica and E. coli, was measured using both semi-batch and continuous cultures bioassays. To assess the ability of CDA combined biocides treatments to remove pre-established biofilms formed on stainless steel discs, CFU counts were performed for both treated and untreated cultures. Eradication of the biofilms by CDA combined antibiotics was evaluated using crystal violet staining. The effect of CDA combined treatments (antibiotics and disinfectants) on biofilm surface area and bacteria viability was evaluated using fluorescence microscopy, digital image analysis and LIVE/DEAD staining. MICs were also determined to assess the probable inhibitory effects of CDA combined treatments on the growth of tested microorganisms' planktonic cells. Treatment of pre-established biofilms with only 310 nM CDA resulted in at least two-fold increase in the number of planktonic cells in all cultures. While antibiotics or disinfectants alone exerted a trivial effect on CFU counts and percentage of surface area covered by the biofilms, combinational treatments with both 310 nM CDA and antibiotics or disinfectants led to approximate 80% reduction in biofilm biomass. These data suggests that combined treatments with CDA would pave the way toward developing new strategies to control biofilms with widespread applications in industry as well as medicine.
Sepehr, Shayesteh; Rahmani-Badi, Azadeh; Babaie-Naiej, Hamta; Soudi, Mohammad Reza
Pre-treatment with EDTA-gallium prevents the formation of biofilms on surfaces
Pseudomonas aeruginosa and Streptococcus pyogenes are leading causes of medical device-associated infections. The capacity to establish and maintain these infections is thought to be associated with the ability to form surface-attached biofilms. In the present study, gallium nitrate was used to coat PVC plates and biofilm formation on the plates by Pseudomonas aeruginosa and Streptococcus pyogenes was evaluated. The results demonstrated that coating the PVC surface with gallium reduced bacteria cell aggregation on the PVC surface and inhibited biofilm formation. These results suggest that surface pre-treatment with a gallium nitrate coating is a potential strategy for the prevention of infections associated with Pseudomonas aeruginosa or Streptococcus pyogenes on medical devices.
Effect of nitrofurans and NO generators on biofilm formation by Pseudomonas aeruginosa PAO1 and Burkholderia cenocepacia 370
Antibacterial drugs in the nitrofuran series, such as nitrofurazone, furazidin, nitrofurantoin and nifuroxazide, as well as the nitric oxide generators sodium nitroprusside and isosorbide mononitrate in concentrations that do not suppress bacterial growth, were shown to increase the capacity of pathogenic bacteria Pseudomonas aeruginosa PAO1 and Burkholderia cenocepacia 370 to form biofilms. At 25�100?g\\/ml, nitrofurans 2�2.5-fold enhanced biofilm formation of
Julia Zaitseva; Vladimir Granik; Alexandr Belik; Olga Koksharova; Inessa Khmel
Biofilm formation and virulence expression by Streptococcus mutans are altered when grown in dual-species model
BACKGROUND: Microbial cell-cell interactions in the oral flora are believed to play an integral role in the development of dental plaque and ultimately, its pathogenicity. The effects of other species of oral bacteria on biofilm formation and virulence gene expression by Streptococcus mutans, the primary etiologic agent of dental caries, were evaluated using a dual-species biofilm model and RealTime-PCR analysis.
Zezhang T. Wen; David Yates; Sang-Joon Ahn; Robert A. Burne
Bacterial Cyclic AMP-Phosphodiesterase Activity Coordinates Biofilm Formation
Biofilm-related infections are a major contributor to human disease, and the capacity for surface attachment and biofilm formation are key attributes for the pathogenesis of microbes. Serratia marcescens type I fimbriae-dependent biofilms are coordinated by the adenylate cyclase, CyaA, and the cyclic 3?,5?-adenosine monophosphate (cAMP)-cAMP receptor protein (CRP) complex. This study uses S. marcescens as a model system to test the role of cAMP-phosphodiesterase activity in controlling biofilm formation. Herein we describe the characterization of a putative S. marcescens cAMP-phosphodiesterase gene (SMA3506), designated as cpdS, and demonstrated to be a functional cAMP-phosphodiesterase both in vitro and in vivo. Deletion of cpdS resulted in defective biofilm formation and reduced type I fimbriae production, whereas multicopy expression of cpdS conferred a type I fimbriae-dependent hyper-biofilm. Together, these results support a model in which bacterial cAMP-phosphodiesterase activity modulates biofilm formation.
Kalivoda, Eric J.; Brothers, Kimberly M.; Stella, Nicholas A.; Schmitt, Matthew J.; Shanks, Robert M. Q.
Flagellar motility is critical for Listeria monocytogenes biofilm formation.
The food-borne pathogen Listeria monocytogenes attaches to environmental surfaces and forms biofilms that can be a source of food contamination, yet little is known about the molecular mechanisms of its biofilm development. We observed that nonmotile mutants were defective in biofilm formation. To investigate how flagella might function during biofilm formation, we compared the wild type with flagellum-minus and paralyzed-flagellum mutants. Both nonmotile mutants were defective in biofilm development, presumably at an early stage, as they were also defective in attachment to glass during the first few hours of surface exposure. This attachment defect could be significantly overcome by providing exogenous movement toward the surface via centrifugation. However, this centrifugation did not restore mature biofilm formation. Our results indicate that it is flagellum-mediated motility that is critical for both initial surface attachment and subsequent biofilm formation. Also, any role for L. monocytogenes flagella as adhesins on abiotic surfaces appears to be either minimal or motility dependent under the conditions we examined. PMID:17416647
Lemon, Katherine P; Higgins, Darren E; Kolter, Roberto
Function of Candida albicans Adhesin Hwp1 in Biofilm Formation
Hwp1 is a well-characterized Candida albicans cell surface protein, expressed only on hyphae, that mediates tight binding to oral epithelial cells. Prior studies indicate that HWP1 expression is dependent upon Bcr1, a key regulator of biofilm formation. Here we test the hypothesis that Hwp1 is required for biofilm formation. In an in vitro model, the hwp1\\/hwp1 mutant produces a thin
Clarissa J. Nobile; Jeniel E. Nett; David R. Andes; Aaron P. Mitchell
Transcriptional Analysis of Biofilm Formation Processes in the Anaerobic, Hyperthermophilic Bacterium Thermotoga maritima
Thermotoga maritima, a fermentative, anaerobic, hyperthermophilic bacterium, was found to attach to bioreactor glass walls, nylon mesh, and polycarbonate filters during chemostat cultivation on maltose-based media at 80�C. A whole-genome cDNA microarray was used to examine differential expression patterns between biofilm and planktonic populations. Mixed-model statistical analysis revealed differential expression (twofold or more) of 114 open reading frames in sessile cells (6% of the genome), over a third of which were initially annotated as hypothetical proteins in the T. maritima genome. Among the previously annotated genes in the T. maritima genome, which showed expression changes during biofilm growth, were several that corresponded to biofilm formation genes identified in mesophilic bacteria (i.e., Pseudomonas species, Escherichia coli, and Staphylococcus epidermidis). Most notably, T. maritima biofilm-bound cells exhibited increased transcription of genes involved in iron and sulfur transport, as well as in biosynthesis of cysteine, thiamine, NAD, and isoprenoid side chains of quinones. These findings were all consistent with the up-regulation of iron-sulfur cluster assembly and repair functions in biofilm cells. Significant up-regulation of several ?-specific glycosidases was also noted in biofilm cells, despite the fact that maltose was the primary carbon source fed to the chemostat. The reasons for increased ?-glycosidase levels are unclear but are likely related to the processing of biofilm-based polysaccharides. In addition to revealing insights into the phenotype of sessile T. maritima communities, the methodology developed here can be extended to study other anaerobic biofilm formation processes as well as to examine aspects of microbial ecology in hydrothermal environments.
Pysz, Marybeth A.; Conners, Shannon B.; Montero, Clemente I.; Shockley, Keith R.; Johnson, Matthew R.; Ward, Donald E.; Kelly, Robert M.
Small Regulatory RNAs in the Control of Motility and Biofilm Formation in E. coli and Salmonella
Biofilm formation in Escherichia coli and other enteric bacteria involves the inverse regulation of the synthesis of flagella and biofilm matrix components such as amyloid curli fibres, cellulose, colanic acid and poly-N-acetylglucosamine (PGA). Physiologically, these processes reflect the transition from growth to stationary phase. At the molecular level, they are tightly controlled by various sigma factors competing for RNA polymerase, a series of transcription factors acting in hierarchical regulatory cascades and several nucleotide messengers, including cyclic-di-GMP. In addition, a surprisingly large number of small regulatory RNAs (sRNAs) have been shown to directly or indirectly modulate motility and/or biofilm formation. This review aims at giving an overview of these sRNA regulators and their impact in biofilm formation in E. coli and Salmonella. Special emphasis will be put on sRNAs, that have known targets such as the mRNAs of the flagellar master regulator FlhDC, the stationary phase sigma factor ?S (RpoS) and the key biofilm regulator CsgD that have recently been shown to act as major hubs for regulation by multiple sRNAs.
Mika, Franziska; Hengge, Regine
Organoselenium Coating on Cellulose Inhibits the Formation of Biofilms by Pseudomonas aeruginosa and Staphylococcus aureus?
Among the most difficult bacterial infections encountered in treating patients are wound infections, which may occur in burn victims, patients with traumatic wounds, necrotic lesions in people with diabetes, and patients with surgical wounds. Within a wound, infecting bacteria frequently develop biofilms. Many current wound dressings are impregnated with antimicrobial agents, such as silver or antibiotics. Diffusion of the agent(s) from the dressing may damage or destroy nearby healthy tissue as well as compromise the effectiveness of the dressing. In contrast, the antimicrobial agent selenium can be covalently attached to the surfaces of a dressing, prolonging its effectiveness. We examined the effectiveness of an organoselenium coating on cellulose discs in inhibiting Pseudomonas aeruginosa and Staphylococcus aureus biofilm formation. Colony biofilm assays revealed that cellulose discs coated with organoselenium completely inhibited P. aeruginosa and S. aureus biofilm formation. Scanning electron microscopy of the cellulose discs confirmed these results. Additionally, the coating on the cellulose discs was stable and effective after a week of incubation in phosphate-buffered saline. These results demonstrate that 0.2% selenium in a coating on cellulose discs effectively inhibits bacterial attachment and biofilm formation and that, unlike other antimicrobial agents, longer periods of exposure to an aqueous environment do not compromise the effectiveness of the coating.
Tran, Phat L.; Hammond, Adrienne A.; Mosley, Thomas; Cortez, Janette; Gray, Tracy; Colmer-Hamood, Jane A.; Shashtri, Mayank; Spallholz, Julian E.; Hamood, Abdul N.; Reid, Ted W.
Studying bacterial hydrophobicity and biofilm formation at liquid-liquid interfaces through interfacial rheology and pendant drop tensiometry.
Bacterial adsorption to interfaces is a key factor in biofilm formation. One major limitation to understanding biofilm formation and development is the accurate measurement of bacterial cell adhesion to hydrophobic interfaces. With this study, bacterial attachment and biofilm growth over time at water-oil interface was monitored through interfacial rheology and tensiometry. Five model bacteria (Pseudomonas putida KT2442, Pseudomonas putida W2, Salmonella typhimurium, Escherichia coli, and Bacillus subtilis) were allowed to adsorb at the water-oil interface either in their non-growing or growing state. We found that we were able to observe the initial kinetics of bacterial attachment and the transient biofilm formation at the water-oil interface through interfacial rheology and tensiometry. Electrophoretic mobility measurements and bacterial adhesion to hydrocarbons (BATH) tests were performed to characterize the selected bacteria. To validate interfacial rheology and tensiometry measurements, we monitored biofilm formation utilizing both confocal laser scanning microscopy and light microscopy. Using this combination of techniques, we were able to observe the elasticity and tension development over time, from the first bacterial attachment up to biofilm formation. PMID:24632390
R�hs, P A; B�cker, L; Inglis, R F; Fischer, P
Two quorum sensing systems control biofilm formation and virulence in members of the Burkholderia cepacia complex
The Burkholderia cepacia complex (Bcc) consists of 17 closely related species that are problematic opportunistic bacterial pathogens for cystic fibrosis patients and immunocompromised individuals. These bacteria are capable of utilizing two different chemical languages: N-acyl homoserine lactones (AHLs) and cis-2-unsaturated fatty acids. Here we summarize the current knowledge of the underlying molecular architectures of these communication systems, showing how they are interlinked and discussing how they regulate overlapping as well as specific sets of genes. A particular focus is laid on the role of these signaling systems in the formation of biofilms, which are believed to be highly important for chronic infections. We review genes that have been implicated in the sessile lifestyle of this group of bacteria. The new emerging role of the intracellular second messenger cyclic dimeric guanosine monophosphate (c-di-GMP) as a downstream regulator of the fatty acid signaling cascade and as a key factor in biofilm formation is also discussed.
Suppiger, Angela; Schmid, Nadine; Aguilar, Claudio; Pessi, Gabriella; Eberl, Leo
Detection of antibiotic-resistant bacteria and their resistance genes in wastewater, surface water, and drinking water biofilms
In view of the increasing interest in the possible role played by hospital and municipal wastewater systems in the selection of antibiotic-resistant bacteria, biofilms were investigated using enterococci, staphylococci, Enterobacteriaceae, and heterotrophic bacteria as indicator organisms. In addition to wastewater, biofilms were also investigated in drinking water from river bank filtrate to estimate the occurrence of resistant bacteria and their
Thomas Schwartz; Wolfgang Kohnen; Bernd Jansen; Ursula Obst
The ?E Pathway Is Involved in Biofilm Formation by Crohn's Disease-Associated Adherent-Invasive Escherichia coli
Ileal lesions of patients with Crohn's disease are colonized by adherent-invasive Escherichia coli (AIEC) bacteria that are able to adhere to and invade intestinal epithelial cells (IEC), to replicate within macrophages, and to form biofilm. Clinical observations showed that bacterial biofilms were associated with the mucosa of inflammatory bowel disease patients. In the present study, we analyzed the relationship between AIEC colonization of the gut and the formation of biofilm, focusing on the involvement of the ?E pathway in the AIEC-IEC interaction. We observed that ?E pathway inhibition in AIEC reference strain LF82 led to an impaired ability to adhere to and invade IEC but also induced a large decrease in the abilities to colonize the intestinal mucosa and form biofilm. This indicates that targeting of the ?E pathway could be a very potent therapeutic strategy by which to interfere with the ability of AIEC to form biofilm on the gut mucosa of Crohn's disease patients.
Chassaing, Benoit
Biofilm formation on enteral feeding tubes by Cronobacter sakazakii, Salmonella serovars and other Enterobacteriaceae.
WHO (2007) recommended that to reduce microbial risks, powdered infant formula should be reconstituted with water at temperatures >70 degrees C, and that such feeds should be used within 2h of preparation. However, this recommendation does not consider the use of enteral feeding tubes which can be in place for more than 48h and can be loci for bacterial attachment. This study determined the extent to which 29 strains of Cronobacter sakazakii, Salmonella serovars, other Enterobacteriaceae and Acinetobacter spp. can adhere and grow on enteral feeding tubes composed of polyvinyl chloride and polyurethane. The study also included silver-impregnated tubing which was expected to have antibacterial activity. Bacterial biofilm formation by members of the Enterobacteriaceae was ca. 10(5)-10(6) cfu/cm after 24h. Negligible biofilm was detected for Acinetobacter gensp. 13; ca. 10 cfu/cm, whereas Cr. sakazakii strain ATCC 12868 had the highest biofilm cell density of 10(7) cfu/cm. Biofilm formation did not correlate with capsule production, and was not inhibited on silver-impregnated tubing. Bacteria grew in the tube lumen to cell densities of 10(7)cfu/ml within 8h, and 10(9)cfu/ml within 24h. It is plausible that in vivo the biofilm will both inoculate subsequent routine feeds and as the biofilm ages, clumps of cells will be shed which may survive passage through the neonate's stomach. Therefore biofilm formation on enteral feeding tubes constitutes a risk factor for susceptible neonates. PMID:19720416
Hurrell, E; Kucerova, E; Loughlin, M; Caubilla-Barron, J; Forsythe, S J
Biofilm formation on brass coupons exposed to a cooling system of an oil refinery
��Brass coupons (70% Cu 30% Zn) were exposed to a cooling freshwater system of an oil refinery, in order to investigate susceptibility\\u000a of the metal to biofilm formation. The coupons were fixed on bypasses at points which allowed the circulation of makeup, cooling\\u000a and return water. The number of aerobic, anaerobic and sulfate-reducing bacteria was determined in both the planktonic
M A N Almeida; F P de Fran�a
Nickel Promotes Biofilm Formation by Escherichia coli K-12 Strains That Produce Curli
The survival of bacteria exposed to toxic compounds is a multifactorial phenomenon, involving well-known molecular mechanisms of resistance but also less-well-understood mechanisms of tolerance that need to be clarified. In particular, the contribution of biofilm formation to survival in the presence of toxic compounds, such as nickel, was investigated in this study. We found that a subinhibitory concentration of nickel
Claire Perrin; Romain Briandet; Gregory Jubelin; Philippe Lejeune; Marie-Andree Mandrand-Berthelot; Agnes Rodrigue; Corinne Dorel
Role of the Clp System in Stress Tolerance, Biofilm Formation, and Intracellular Invasion in Porphyromonas gingivalis
Clp proteases and chaperones are ubiquitous among prokaryotes and eukaryotes, and in many pathogenic bacteria the Clp stress response system is also involved in regulation of virulence properties. In this study, the roles of ClpB, ClpC, and ClpXP in stress resistance, homotypic and heterotypic biofilm formation, and intracellular invasion in the oral opportunistic pathogen Porphyromonas gingivalis were investigated. Absence of
Cindy A. Capestany; Gena D. Tribble; Kazuhiko Maeda; Donald R Demuth; Richard J. Lamont
Action of antimicrobial substances produced by different oil reservoir Bacillus strains against biofilm formation
Microbial colonization of petroleum industry systems takes place through the formation of biofilms, and can result in biodeterioration\\u000a of the metal surfaces. In a previous study, two oil reservoir Bacillus strains (Bacillus licheniformis T6-5 and Bacillus firmus H2O-1) were shown to produce antimicrobial substances (AMS) active against different Bacillus strains and a consortium of sulfate-reducing bacteria (SRB) on solid medium.
E. Korenblum; G. V. Sebasti�n; M. M. Paiva; C. M. L. M. Coutinho; F. C. M. Magalh�es; B. M. Peyton; L. Seldin
Biofilm formation and composition on different implant materials in vivo.
Biofilm formation was evaluated on the following titanium and zirconia implants in vivo: machined titanium (Ti-m), modified titanium (TiUnite), modified zirconia (ZiUnite), machined alumina-toughened zirconia (ATZ-m), sandblasted alumina-toughened zirconia (ATZ-s), and machined zirconia (TZP-A-m). Bovine enamel slabs were used as controls. Surface morphologies were examined by atomic force (AFM) and scanning electron microscopy (SEM). The surface wettability was also determined. Twelve healthy volunteers wore a splint system with the tested materials. After 3 and 5 days the materials were examined by fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy (CLSM). The levels of Streptococcus spp., Veillonella spp., Fusobacteriaum nucleatum, and Actinomyces naeslundii were quantitatively determined. The biofilm thickness was found to be between 19.78 and 36.73 ?m after 3 days and between 26.11 and 32.43 ?m after 5 days. With the exception of Ti-m the biofilm thickness after 3 days was correlated with surface roughness. In addition to Streptococcus spp. as the main component of the biofilm (11.23-25.30%), F. nucleatum, A. naeslundii, and Veillonella spp. were also detected. No significant differences in biofilm composition on the implant surfaces could be observed. In total, the influence of roughness and material on biofilm formation was compensated by biofilm maturation. PMID:20725954
Al-Ahmad, A; Wiedmann-Al-Ahmad, M; Faust, J; B�chle, M; Follo, M; Wolkewitz, M; Hannig, C; Hellwig, E; Carvalho, C; Kohal, R
The EpsE Flagellar Clutch Is Bifunctional and Synergizes with EPS Biosynthesis to Promote Bacillus subtilis Biofilm Formation
Many bacteria inhibit motility concomitant with the synthesis of an extracellular polysaccharide matrix and the formation of biofilm aggregates. In Bacillus subtilis biofilms, motility is inhibited by EpsE, which acts as a clutch on the flagella rotor to inhibit motility, and which is encoded within the 15 gene eps operon required for EPS production. EpsE shows sequence similarity to the glycosyltransferase family of enzymes, and we demonstrate that the conserved active site motif is required for EPS biosynthesis. We also screen for residues specifically required for either clutch or enzymatic activity and demonstrate that the two functions are genetically separable. Finally, we show that, whereas EPS synthesis activity is dominant for biofilm formation, both functions of EpsE synergize to stabilize cell aggregates and relieve selective pressure to abolish motility by genetic mutation. Thus, the transition from motility to biofilm formation may be governed by a single bifunctional enzyme.
Guttenplan, Sarah B.; Blair, Kris M.; Kearns, Daniel B.
Impact of Early Colonizers on In Vitro Subgingival Biofilm Formation
The aim of this study was to investigate the impact of early colonizing species on the structure and the composition of the bacterial community developing in a subgingival 10-species biofilm model system. The model included Streptococcus oralis, Streptococcus anginosus, Actinomycesoris, Fusobacterium nucleatum subsp. nucleatum, Veillonella dispar, Campylobacter rectus, Prevotella intermedia, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola. Based on literature, we considered Streptococcus oralis, Streptococcus anginosus, and Actinomyces oris as early colonizers and examined their role in the biofilms by either a delayed addition to the consortium, or by not inoculating at all the biofilms with these species. We quantitatively evaluated the resulting biofilms by real-time quantitative PCR and further compared the structures using confocal laser scanning microscopy following fluorescence in situ hybridisation. The absence of the early colonizers did not hinder biofilm formation. The biofilms reached the same total counts and developed to normal thickness. However, quantitative shifts in the abundances of individual species were observed. In the absence of streptococci, the overall biofilm structure appeared looser and more dispersed. Moreover, besides a significant increase of P. intermedia and a decrease of P. gingivalis , P. intermedia appeared to form filamented long chains that resembled streptococci. A. oris, although growing to significantly higher abundance in absence of streptococci, did not have a visible impact on the biofilms. Hence, in the absence of the early colonizers, there is a pronounced effect on P. intermedia and P. gingivalis that may cause distinct shifts in the structure of the biofilm. Streptococci possibly facilitate the establishment of P. gingivalis into subgingival biofilms, while in their absence P. intermedia became more dominant and forms elongated chains.
Ammann, Thomas W.; Belibasakis, Georgios N.; Thurnheer, Thomas
Cyclic-di-GMP signalling regulates motility and biofilm formation in Bordetella bronchiseptica
The signalling molecule bis-(3?�5?)-cyclic-dimeric guanosine monophosphate (c-di-GMP) is a central regulator of diverse cellular functions, including motility, biofilm formation, cell cycle progression and virulence, in bacteria. Multiple diguanylate cyclase and phosphodiesterase-domain-containing proteins (GGDEF and EAL/HD-GYP, respectively) modulate the levels of the second messenger c-di-GMP to transmit signals and obtain such specific cellular responses. In the genus Bordetella this c-di-GMP network is poorly studied. In this work, we evaluated the expression of two phenotypes in Bordetella bronchiseptica regulated by c-di-GMP, biofilm formation and motility, under the influence of ectopic expression of Pseudomonas aeruginosa proteins with EAL or GGDEF domains that regulates the c-di-GMP level. In agreement with previous reports for other bacteria, we observed that B. bronchiseptica is able to form biofilm and reduce its motility only when GGDEF domain protein is expressed. Moreover we identify a GGDEF domain protein (BB3576) with diguanylate cyclase activity that participates in motility and biofilm regulation in B. bronchiseptica. These results demonstrate for the first time, to our knowledge, the presence of c-di-GMP regulatory signalling in B. bronchiseptica.
Sisti, Federico; Ha, Dae-Gon; O'Toole, George A.; Hozbor, Daniela
Quorum sensing controls biofilm formation in Vibrio cholerae through modulation of cyclic di-GMP levels and repression of vpsT.
Two chemical signaling systems, quorum sensing (QS) and 3',5'-cyclic diguanylic acid (c-di-GMP), reciprocally control biofilm formation in Vibrio cholerae. QS is the process by which bacteria communicate via the secretion and detection of autoinducers, and in V. cholerae, QS represses biofilm formation. c-di-GMP is an intracellular second messenger that contains information regarding local environmental conditions, and in V. cholerae, c-di-GMP activates biofilm formation. Here we show that HapR, a major regulator of QS, represses biofilm formation in V. cholerae through two distinct mechanisms. HapR controls the transcription of 14 genes encoding a group of proteins that synthesize and degrade c-di-GMP. The net effect of this transcriptional program is a reduction in cellular c-di-GMP levels at high cell density and, consequently, a decrease in biofilm formation. Increasing the c-di-GMP concentration at high cell density to the level present in the low-cell-density QS state restores biofilm formation, showing that c-di-GMP is epistatic to QS in the control of biofilm formation in V. cholerae. In addition, HapR binds to and directly represses the expression of the biofilm transcriptional activator, vpsT. Together, our results suggest that V. cholerae integrates information about the vicinal bacterial community contained in extracellular QS autoinducers with the intracellular environmental information encoded in c-di-GMP to control biofilm formation. PMID:18223081
Waters, Christopher M; Lu, Wenyun; Rabinowitz, Joshua D; Bassler, Bonnie L
MgrA Represses Biofilm Formation in Staphylococcus aureus
MgrA is a pleiotropic regulator that controls autolysis, virulence, and efflux pump activity in Staphylococcus aureus. We recently found that mgrA mutants of strains RN6390, SH1000, and MW2 also displayed enhanced biofilm formation compared with their respective parents. The biofilms formed by mgrA mutants of RN6390 and MW2 are independent of sigB and ica loci, two genetic elements that have
Mar�a Pilar Trotonda; Sandeep Tamber; Guido Memmi; Ambrose L. Cheung
Extracellular polysaccharides produced by cooling water tower biofilm bacteria and their possible degradation
The extracellular polymers (EPS) of biofilm bacteria that can cause heat and mass transfer problems in cooling water towers in the petrochemical industry were investigated. In addition, these microorganisms were screened for their ability to grow and degrade their own EPS and the EPS of other species. Twelve bacteria producing the most EPS were isolated from cooling water towers and
Nur Ceyhan; Guven Ozdemir
Nanostructured selenium for preventing biofilm formation on polycarbonate medical devices.
Biofilms are a common cause of persistent infections on medical devices as they are easy to form and hard to treat. The objective of this study was for the first time to coat selenium (a natural element in the body) nanoparticles on the surface of polycarbonate medical devices (such as those used for medical catheters) and to examine their effectiveness at preventing biofilm formation. The size and distribution of selenium coatings were characterized using scanning electron microscopy and atomic force microscopy. The strength of the selenium coating on polycarbonate was assessed by tape-adhesion tests followed by atomic absorption spectroscopy. Results showed that selenium nanoparticles had a diameter of 50-100 nm and were well distributed on the polycarbonate surface. In addition, more than 50% of the selenium coating survived the tape-adhesion test as larger nanoparticles had less adhesion strength to the underlying polycarbonate substrate than smaller selenium nanoparticles. Most significantly, the results of this in vitro study showed that the selenium coatings on polycarbonate significantly inhibited Staphylococcus aureus growth to 8.9% and 27% when compared with an uncoated polycarbonate surface after 24 and 72 h, respectively. Importantly, this was accomplished without using antibiotics but rather with an element (selenium) that is natural to the human body. Thus, this study suggests that coating polymers (particularly, polycarbonate) with nanostructured selenium is a fast and effective way to reduce bacteria functions that lead to medical device infections. � 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 100A: 3205-3210, 2012. PMID:22707390
Wang, Qi; Webster, Thomas J
Variation in sessile microflora during biofilm formation on AISI-304 stainless steel coupons.
Coupons of stainless steel type AISI-304 were exposed to the industrial cooling system of a petrochemical plant fed by seawater from the Guanabara Bay, Rio de Janeiro, Brazil, in order to study the in situ formation of biofilms. Bacteria, microalgae and fungi were detected on the coupons as soon as 48 h after exposure. Their respective numbers were determined at times 48, 96 and 192 h and over the following 8 weeks. Aerobic, anaerobic and sulfate-reducing bacteria were quantified according to the technique of the most probable number, and fungi by the pour plate technique. The number of microorganisms present in the forming biofilm varied over the experimental period, reaching maximal levels of 14 x 10(11) cells cm-2, 30 x 10(13) cells cm-2, 38 x 10(11) cells cm-2 and 63 x 10(5) cells cm-2, respectively, for aerobic bacteria, anaerobic bacteria, sulfate-reducing bacteria and fungi, and the dynamics of this variation depended on the group of microorganisms. Bacillus sp, Escherichia coli, Serratia sp and Pseudomonas putrefaciens were identified among the aerobic bacteria isolated. Additionally, microalgae and bacteria of the genus Gallionella were also detected. Nonetheless, no evidence of corrosion was found on the stainless steel type AISI-304 coupons over the experimental period. PMID:8987686
de Fran�a, F P; Lutterbach, M T
Role of the Clp System in Stress Tolerance, Biofilm Formation, and Intracellular Invasion in Porphyromonas gingivalis?
Clp proteases and chaperones are ubiquitous among prokaryotes and eukaryotes, and in many pathogenic bacteria the Clp stress response system is also involved in regulation of virulence properties. In this study, the roles of ClpB, ClpC, and ClpXP in stress resistance, homotypic and heterotypic biofilm formation, and intracellular invasion in the oral opportunistic pathogen Porphyromonas gingivalis were investigated. Absence of ClpC and ClpXP, but not ClpB, resulted in diminished tolerance to high temperatures. Response to oxidative stress was not affected by the loss of any of the Clp proteins. The clpC and clpXP mutants demonstrated elevated monospecies biofilm formation, and the absence of ClpXP also enhanced heterotypic P. gingivalis-Streptococcus gordonii biofilm formation. All clp mutants adhered to gingival epithelial cells to the same level as the wild type; however, ClpC and ClpXP were found to be necessary for entry into host epithelial cells. ClpB did not play a role in entry but was required for intracellular replication and survival. ClpXP negatively regulated the surface exposure of the minor fimbrial (Mfa) protein subunit of P. gingivalis, which stimulates biofilm formation but interferes with epithelial cell entry. Collectively, these results show that the Clp protease complex and chaperones control several processes that are important for the colonization and survival of P. gingivalis in the oral cavity.
Capestany, Cindy A.; Tribble, Gena D.; Maeda, Kazuhiko; Demuth, Donald R.; Lamont, Richard J.
Functional Analysis of the Protein Veg, Which Stimulates Biofilm Formation in Bacillus subtilis
Biofilm is a complex aggregate of cells that adhere to each other and produce an extracellular matrix. In Bacillus subtilis, an extracellular polysaccharide (EPS) and amyloid fiber (TasA), synthesized by the epsA-epsO and tapA-sipW-tasA operons, respectively, are the primary components of the extracellular matrix. In the current study, we investigated the functional role of the previously uncharacterized veg gene in B. subtilis. Overproduction of Veg, a small protein highly conserved among Gram-positive bacteria, stimulated biofilm formation via inducing transcription of the tapA-sipW-tasA operon. Moreover, overproduced Veg restored the impairment of biofilm formation in mutants carrying a deletion of of sinI, slrA, or slrR, encoding an antirepressor of SinR that acts as the master regulator of biofilm formation, while biofilm morphology in the absence of SinR was not affected by either additional veg deletion or overproduction, indicating that Veg negatively regulates SinR activity independently of the known antirepressors. Expression of sinR was not affected in Veg-overproducing cells, and amounts of SinR were similar in cells expressing different levels of Veg, strongly suggesting that Veg modulates the repressor activity of SinR. Interestingly, the results of in vivo pulldown assays of the SinR complex indicate that Veg inhibits the interactions between SinR and SlrR. Based on these findings, we propose that Veg or a Veg-induced protein acts as an antirepressor of SinR to regulate biofilm formation.
Lei, Ying; Oshima, Taku; Ogasawara, Naotake
A new synthetic ligand that activates QscR and blocks antibiotic-tolerant biofilm formation in Pseudomonas aeruginosa.
Quorum sensing (QS) has been recognized to play an important role in many pathogenic bacteria and has become a novel target for the treatment of infectious disease. Pseudomonas aeruginosa is highly resistant to antibiotic treatment, largely due to its ability to form biofilms, and QS was found to be essential for the creation of mature, differentiated biofilms in this organism. A novel QS inhibitor, C2 (N-decanoyl-L-homoserine benzyl ester), can attenuate not only total protease and elastase activity, but also swarming motility and biofilm formation in the P. aeruginosa strain PAO1. We demonstrated that C2 showed a significant inhibitory effect on biofilm formation in a dose-dependent manner. Data from cDNA microarray showed that expression of 382 genes (?6.4 %) was significantly different with C2 treatment, including downregulation of 215 genes (?3.6 %) and upregulation of 167 genes (?2.8 %). Real-time reverse transcription-polymerase chain reaction (RT-PCR) showed that the gene qscR, which encodes the LuxR-type receptor QscR (quorum sensing control repressor), was significantly upregulated by 375.4 % during C2 treatment. The mechanism by which C2 inhibits biofilm formation may be through repression of Las and Rhl systems by QscR. C2 was shown to reduce biofilm formation; in combination with antibiotics, it abolishes biofilm formation completely. This result may pave the way for new treatments for biofilm-related infections and may be exploited for the general prevention of biofilm formation. PMID:24327212
Weng, Li-Xing; Yang, Yu-Xiang; Zhang, Yu-Qian; Wang, Lian-Hui
Biofilm streamer formation in a microfluidic porous media mimic
NASA Astrophysics Data System (ADS)
Biofilm formation in porous media is of significant importance in many environmental and industrial processes such as bioremediation, oil recovery, and wastewater treatment. Among different biological and environmental factors, hydrodynamics is considered an important determinant of the dynamics of biofilm formation. In the present study, we fabricated a microfluidic porous media mimic and investigated how fluid flow influences the formation of filamentous structures, known as streamers, between porous media structures. Streamers are viscoelastic materials composed of extracellular polymeric substances (EPS) and bacterial cells, and these filamentous structures are typically tethered at either one of both ends to surfaces. We studied evolution of streamers in different flow rates and identified a tangible link between hydrodynamic conditions and development of these filamentous structures. Our results show that hydrodynamic conditions not only determine the limit of the streamers formation, but also influence both temporal evolution and spatial organization of biofilm streamers.
Kumar, Aloke; Valiei, Amin; Mukherjee, Partha; Liu, Yang; Thundat, Thomas
Brominated Furanones Inhibit Biofilm Formation by Salmonella enterica Serovar Typhimurium ?
Salmonella enterica serovar Typhimurium is a main cause of bacterial food-borne diseases. As Salmonella can form biofilms in which it is better protected against antimicrobial agents on a wide diversity of surfaces, it is of interest to explore ways to inhibit biofilm formation. Brominated furanones, originally extracted from the marine alga Delisea pulchra, are known to interfere with biofilm formation in several pathogens. In this study, we have synthesized a small focused library of brominated furanones and tested their activity against S. enterica serovar Typhimurium biofilm formation. We show that several furanones inhibit Salmonella biofilm formation at non-growth-inhibiting concentrations. The most interesting compounds are (Z)-4-bromo-5-(bromomethylene)-3-alkyl-2(5H)-furanones with chain lengths of two to six carbon atoms. A microarray study was performed to analyze the gene expression profiles of Salmonella in the presence of (Z)-4-bromo-5-(bromomethylene)-3-ethyl-2(5H)-furanone. The induced genes include genes that are involved in metabolism, stress response, and drug sensitivity. Most of the repressed genes are involved in metabolism, the type III secretion system, and flagellar biosynthesis. Follow-up experiments confirmed that this furanone interferes with the synthesis of flagella by Salmonella. No evidence was found that furanones act on the currently known quorum-sensing systems in Salmonella. Interestingly, pretreatment with furanones rendered Salmonella biofilms more susceptible to antibiotic treatment. Conclusively, this work demonstrates that particular brominated furanones have potential in the prevention of biofilm formation by Salmonella serovar Typhimurium.
Janssens, Joost C. A.; Steenackers, Hans; Robijns, Stijn; Gellens, Edith; Levin, Jeremy; Zhao, Hui; Hermans, Kim; De Coster, David; Verhoeven, Tine L.; Marchal, Kathleen; Vanderleyden, Jos; De Vos, Dirk E.; De Keersmaecker, Sigrid C. J.
Cattle tick-associated bacteria exert anti-biofilm and anti-Tritrichomonas foetus activities.
Research on microbiota in cattle tick and the evaluation of its activity against other microorganisms can contribute to identify new molecules potentially useful to control infections caused by bacteria and protozoa. Biofilms pose increasing problems worldwide, mainly due to their resistance to antimicrobial therapies and host immune response. In this study we investigate the ability Rhipicephalus (Boophilus) microplus-associated bacteria may exhibit to produce anti-biofilm and trichomonicidal compounds. Gut, ovary, salivary glands, and Gen� organ were collected from engorged R. microplus female. Homogenates of each tissue were inoculated onto 15 distinct culture media. Anti-biofilm and trichomonicidal activities were analyzed by culturing each bacterium isolated in a liquid medium. Results showed that R. microplus cattle tick microflora varies for different tissues. Bacteria belonging to different genera (Aeromonas, Bacillus, Brevibacillus, Castelaniella, Comamonas, Kocuria, and Microbacterium) were identified. Interestingly, all bacterial species found displayed pronounced activity against Staphylococcus epidermidis and Pseudomonas aeruginosa biofilms, and also against the cattle pathogen Tritrichomonas foetus, confirming the hypothesis that cattle tick could be a source of bacteria active against pathogens. This is the first study showing that bacteria isolated from a tick exert anti-biofilm and trichomonicidal activities. PMID:23434012
Zimmer, K R; Seixas, A; Concei��o, J M; Zvoboda, D A; Barros, M P; Tasca, T; Macedo, A J; Termignoni, C
Enzymatic catalysis of mercury methylation by planktonic and biofilm cultures of sulfate- reducing bacteria
NASA Astrophysics Data System (ADS)
While biofilms are now known to be the predominant form of microbial growth in nature, little is known about their role in environmental mercury (Hg) methylation. Due to its long-range atmospheric transport, Hg contamination of food chains is a worldwide problem, impacting even pristine areas. Among different forms of mercury species, methylmercury (MeHg) is an extremely neurotoxic and biomagnification-prone compound that can lead to severely adverse health effects on wildlife and humans. Considerable studies have shown that in the aquatic environment the external supply of MeHg is not sufficient to account for MeHg accumulation in biota and in situ biological MeHg formation plays a critical role in determining the amount of MeHg in food webs; moreover, sulfate-reducing bacteria (SRB) has been identified as the principal Hg-methylating organisms in nature. In a wide range of aquatic systems wetlands are considered important sites for Hg methylation mostly because of the environmental factors that promote microbial activity within, and biofilms are especially important in wetland ecosystems due to large amount of submerged surfaces. Although recent work has focused on the environmental factors that control MeHg production and the conditions that affect the availability of inorganic Hg to SRB, much remains to be understood about the biochemical mechanism of the Hg methylation process in SRB, especially in the biofilm-growth of these microbes. Data from our previous study with SRB strains isolated from a coastal wetland suggested that the specific Hg methylation rate found was approximately an order of magnitude higher in biofilm cells than in planktonic cells. In order to investigate possible reasons for this observed difference, and to test if this phenomenon is observed in other strains, we conducted chloroform, fluroacetate and molybdate inhibition assays in both complete and incomplete-oxidizing SRB species (Desulfovibrio desulfuricans M8, Desulfococcus sp. Desulfobactor sp. BG8) grown in planktonic and biofilm form, as the acetyl- coenzyme A pathway involved with cobalamin has been hypothesized to be the pathway for Hg methylation. The purpose of this study was to probe whether differences in the enzymatically catalyzed process caused differential methylation rates between the species and also between the different forms of culture growth. Any attempts to control the environmentally undesirable Hg methylation process would benefit from a better understanding of the biochemical mechanism involved.
Lin, C.; Kampalath, R.; Jay, J.
Resistance of Pseudomonas aeruginosa to liquid disinfectants on contaminated surfaces before formation of biofilms.
A comparison was made of the effectiveness of popular disinfectants (Cavicide, Cidexplus, Clorox, Exspor, Lysol, Renalin, and Wavicide) under conditions prescribed for disinfection in the respective product labels on Pseudomonas aeruginosa either in suspension or deposited onto surfaces of metallic or polymeric plastic devices. The testing also included 7 nonformulated germicidal agents (glutaraldehyde, formaldehyde, peracetic acid, hydrogen peroxide, sodium hypochlorite, phenol, and cupric ascorbate) commonly used in disinfection and decontamination. Results showed that P. aeruginosa is on average 300-fold more resistant when present on contaminated surfaces than in suspension. This increase in resistance agrees with results reported in studies of biofilms, but unexpectedly, it precedes biofilm formation. The surface to which bacteria are attached can influence the effectiveness of disinfectants. Viable bacteria attached to devices may require dislodging through more than a one-step method for detection. The data, obtained with a sensitive and quantitative test, suggest that disinfectants are less effective on contaminated surfaces than generally acknowledged. PMID:11128146
Sagripanti, J L; Bonifacino, A
Fibronectin-binding proteins are required for biofilm formation by community-associated methicillin-resistant Staphylococcus aureus strain LAC.
Community-associated methicillin-resistant Staphylococcus aureus of the USA300 lineage is emerging as an important cause of medical device-related infection. However, few factors required for biofilm accumulation by USA300 strains have been identified, and the processes involved are poorly understood. Here, we identify S.�aureus proteins required for the USA300 isolate LAC to form biofilm. A mutant with a deletion of the fnbA and fnbB genes did not express the fibronectin-binding proteins FnBPA and FnBPB and lacked the ability to adhere to fibronectin or to form biofilm. Biofilm formation by the mutant LAC?fnbAfnbB could be restored by expression of FnBPA or FnBPB from a plasmid demonstrating that both of these proteins can mediate biofilm formation when expressed by LAC. Expression of FnBPA and FnBPB increased bacterial aggregation suggesting that fibronectin-binding proteins can promote the accumulation phase of biofilm. Loss of fibronectin-binding proteins reduced the initial adherence of bacteria, indicating that these proteins are also involved in primary attachment. In summary, these findings improve our understanding of biofilm formation by the USA300 strain LAC by demonstrating that the fibronectin-binding proteins are required. PMID:24628034
McCourt, Jennifer; O'Halloran, Dara P; McCarthy, Hannah; O'Gara, James P; Geoghegan, Joan A
Bap, a Staphylococcus aureus Surface Protein Involved in Biofilm Formation
Identification of new genes involved in biofilm formation is needed to understand the molecular basis of strain variation and the pathogenic mechanisms implicated in chronic staphylococcal infections. A biofilm-producing Staphylococcus aureus isolate was used to generate biofilm-negative transposon (Tn917) insertion mutants. Two mutants were found with a significant decrease in attachment to inert surfaces (early adherence), intercellular adhesion, and biofilm formation. The transposon was inserted at the same locus in both mutants. This locus (bap [for biofilm associated protein]) encodes a novel cell wall associated protein of 2,276 amino acids (Bap), which shows global organizational similarities to surface proteins of gram-negative (Pseudomonas aeruginosa and Salmonella enterica serovar Typhi) and gram-positive (Enteroccocus faecalis) microorganisms. Bap's core region represents 52% of the protein and consists of 13 successive nearly identical repeats, each containing 86 amino acids. bap was present in a small fraction of bovine mastitis isolates (5% of the 350 S. aureus isolates tested), but it was absent from the 75 clinical human S. aureus isolates analyzed. All staphylococcal isolates harboring bap were highly adherent and strong biofilm producers. In a mouse infection model bap was involved in pathogenesis, causing a persistent infection.
Cucarella, Carme; Solano, Cristina; Valle, Jaione; Amorena, Beatriz; Lasa, Inigo; Penades, Jose R.
A Differential Effect of E. coli Toxin-Antitoxin Systems on Cell Death in Liquid Media and Biofilm Formation
Toxin-antitoxin (TA) modules are gene pairs specifying for a toxin and its antitoxin and are found on the chromosomes of many bacteria including pathogens. Here we report how each of five such TA systems in E. coli affect bacterial cell death differently in liquid media and during biofilm formation. Of all these systems, only the TA system mazEF mediated cell
Ilana Kolodkin-Gal; Reut Verdiger; Ayalla Shlosberg-Fedida; Hanna Engelberg-Kulka; Stefan Bereswill
Effects of Carbon Source, Carbon Concentration, and Chlorination on Growth Related Parameters of Heterotrophic Biofilm Bacteria
To investigate growth of heterotrophic biofilm bacteria, a model biofilm reactor was developed to simulate a drinking water\\u000a distribution system. Controlled addition of three different carbon sources (amino acids, carbohydrates, and humics) at three\\u000a different concentrations (500, 1,000, and 2,000 ppb carbon) in the presence and absence of chlorine were used in separate\\u000a experiments. An additional experiment was run with
B. D. Ellis; P. Butterfield; W. L. Jones; G. A. McFeters; A. K. Camper
Virulence regulator PrfA is essential for biofilm formation in Listeria monocytogenes but not in Listeria innocua.
The ability of the foodborne pathogen Listeria monocytogenes to develop biofilm in food-processing environment is a major concern for the food safety, because biofilms allow bacteria to better resist environmental stresses. PrfA is a key transcriptional activator that positively regulates most of the known listerial virulence gene expression. In order to explore the role of PrfA on Listeria biofilm development, we compared the abilities of biofilm formation by L. monocytogenes wild type strains (EGD and EGDe) and their prfA deletion mutants (EGD?prfA and EGDe?prfA), nonpathogenic Listeria innocua, as well as the recombinant strains that express constitutively active mutant PrfA (PrfA*) in L. innocua (LI-pERL3-prfA*) and in EGDe?prfA (EGDe?prfA-pERL3-prfA*) at 37�C in brain heart infusion (BHI) medium using the polyvinyl chloride (PVC) microtiter plate assay and microscopic examination. Our results showed that the wild types of L. monocytogenes had strong abilities to develop biofilm with meshwork of bacterial aggregates, while biofilm with sparse small clumps were observed in L. innocua. The biofilm production of strains EGD?prfA and EGDe?prfA that lack funtional PrfA was reduced and could be recovered by the introduction of the PrfA*, however, the PrfA* had no impact on the biofilm forming ability of L. innocua. Our results suggest that PrfA plays a significant role in biofilm formation in L. monocytogenes but not in L. innocua, thus may reflect differences in the molecular mechanisms of biofilm formation by these two closely related species. PMID:21656247
Zhou, Qingchun; Feng, Feifei; Wang, Li; Feng, Xiaoqin; Yin, Xiaojiao; Luo, Qin
Biofilm formation by the yeast Rhodotorula mucilaginosa: process, repeatability and cell attachment in a continuous biofilm reactor.
Yeast biofilms contribute to quality impairment of industrial processes and also play an important role in clinical infections. Little is known about biofilm formation and their treatment. The aim of this study was to establish a multi-layer yeast biofilm model using a modified 3.7 l bench-top bioreactor operated in continuous mode (D = 0.12 h(-1)). The repeatability of biofilm formation was tested by comparing five bioprocesses with Rhodotorula mucilaginosa, a strain isolated from washing machines. The amount of biofilm formed after 6 days post inoculation was 83 ?g cm(-2) protein, 197 ?g cm(-2) polysaccharide and 6.9 � 10(6) CFU cm(-2) on smooth polypropylene surfaces. Roughening the surface doubled the amount of biofilm but also increased its spatial variability. Plasma modification of polypropylene significantly reduced the hydrophobicity but did not enhance cell attachment. The biofilm formed on polypropylene coupons could be used for sanitation studies. PMID:21981116
Gattlen, Jasmin; Zinn, Manfred; Guimond, S�bastien; K�rner, Enrico; Amberg, Caroline; Mauclaire, Laurie
The impact of influent total ammonium nitrogen concentration on nitrite-oxidizing bacteria inhibition in moving bed biofilm reactor.
The application of nitrification-denitrification over nitrite (nitritation-denitritation) with municipal (i.e. diluted and cold (or low-temperature)) wastewater can substantially improve the energy balance of municipal wastewater treatment plants. For the accumulation of nitrite, it is crucial to inhibit nitrite-oxidizing bacteria (NOB) with simultaneous proliferation of ammonium-oxidizing bacteria (AOB). The present study describes the effect of the influent total ammonium nitrogen (TAN) concentration on AOB and NOB activity in two moving bed biofilm reactors operated as sequencing batch reactors (SBR) at 15 �C (SBR I) and 21 �C (SBR II). The reactors were fed with diluted reject water containing 600, 300, 150 and 75 mg TAN L(-1). The only factor limiting NOB activity in these reactors was the high concentrations of free ammonia and/or free nitrous acid (FNA) during the SBR cycles. Nitrite accumulation was observed with influents containing 600, 300 and 150 mg TAN L(-1) in SBR I and 600 and 300 in SBR II. Once nitrate production established in the reactors, the increase of influent TAN concentration up to the original 600 mg TAN L(-1) did not limit NOB activity. This was due to the massive development of NOB clusters throughout the biofilm that were able to cope with faster formation of FNA. The results of the fluorescence in situ hybridization analysis preliminarily showed the stratification of bacteria in the biofilm. PMID:24647188
Kouba, Vojtech; Catrysse, Michael; Stryjova, Hana; Jonatova, Ivana; Volcke, Eveline I P; Svehla, Pavel; Bartacek, Jan
Newly-synthesized chalcones-inhibition of adherence and biofilm formation of methicillin-resistant Staphylococcus aureus
Biofilm formation and adherence of bacteria to host tissue are one of the most important virulence factors of methicillin-resistant strains of Staphylococcus aureus (MRSA). The number of resistant strains is seriously increasing during the past years and bacteria have become resistant, not only to methicillin, but also to other commonly used antistaphylococcal antibiotics. There is a great need for discovering a novel antimicrobial agent for the treatment of staphylococcal infections. One of the most promising groups of compounds appears to be chalcones. In present study we evaluated the in vitro effect of three newly synthesized chalcones: 1,3- Bis-(2-hydroxy-phenyl)-propenone, 3-(3-Hydroxy-phenyl)-1-(2-hydroxy-phenyl)-propenone and 3-(4-Hydroxy-phenyl)-1-(2-hydroxy-phenyl)-propenone on glycocalyx production, biofilm formation and adherence to human fibronectin of clinical isolates and laboratory control strain of MRSA (ATCC 43300). Subinhibitory concentrations of the tested compounds reduced the production of glycocalyx, biofilm formation and adherence to human fibronectin of all MRSA strains. Inhibition of biofilm formation was dose dependent and the most effective was 1,3- Bis-(2-hydroxy-phenyl)-propenone. In our study we demonstrated that three newly-synthesized chalcones exhibited significant effect on adherence and biofilm formation of MRSA strains. Chalcones may be considered as promising new antimicrobial agents that can be used for prevention of staphylococcal infections or as adjunct to antibiotics in conventional therapy.
Bozic, Dragana D.; Milenkovic, Marina; Ivkovic, Branka; Cirkovic, Ivana
Newly-synthesized chalcones-inhibition of adherence and biofilm formation of methicillin-resistant Staphylococcus aureus.
Biofilm formation and adherence of bacteria to host tissue are one of the most important virulence factors of methicillin-resistant strains of Staphylococcus aureus (MRSA). The number of resistant strains is seriously increasing during the past years and bacteria have become resistant, not only to methicillin, but also to other commonly used antistaphylococcal antibiotics. There is a great need for discovering a novel antimicrobial agent for the treatment of staphylococcal infections. One of the most promising groups of compounds appears to be chalcones. In present study we evaluated the in vitro effect of three newly synthesized chalcones: 1,3- Bis-(2-hydroxy-phenyl)-propenone, 3-(3-Hydroxy-phenyl)-1-(2-hydroxy-phenyl)-propenone and 3-(4-Hydroxy-phenyl)-1-(2-hydroxy-phenyl)-propenone on glycocalyx production, biofilm formation and adherence to human fibronectin of clinical isolates and laboratory control strain of MRSA (ATCC 43300). Subinhibitory concentrations of the tested compounds reduced the production of glycocalyx, biofilm formation and adherence to human fibronectin of all MRSA strains. Inhibition of biofilm formation was dose dependent and the most effective was 1,3- Bis-(2-hydroxy-phenyl)-propenone. In our study we demonstrated that three newly-synthesized chalcones exhibited significant effect on adherence and biofilm formation of MRSA strains. Chalcones may be considered as promising new antimicrobial agents that can be used for prevention of staphylococcal infections or as adjunct to antibiotics in conventional therapy. PMID:24948943
Bozic, Dragana D; Milenkovic, Marina; Ivkovic, Branka; Cirkovic, Ivana
Biofilm formation on abiotic and biotic surfaces during Spanish style green table olive fermentation.
In this work, the establishment of polymicrobial communities on the surfaces which come into contact with the brine during Spanish style Gordal cv. green olive fermentation when subjected to spontaneous or controlled processes (inoculated with Lactobacillus pentosus LPCO10 or 128/2) was studied. Scanning electron microscopy showed that L. pentosus and yeast populations were able to form mixed biofilms throughout the fermentation process on both abiotic (glass slide) and biotic (olive skin) surfaces. The biofilm architectures in both supports were completely different: on the glass slides only aggregates of L. pentosus and yeasts without any polymeric matrix surrounding them were found while on the skin of the fruits, true mature biofilms were observed. During fermentation, the lactic acid bacteria (LAB) population on the olives remained similar while that of yeasts increased progressively to reach similar levels at the end of the process (8-9 log CFU/cm(2)). Molecular analysis showed that different populations of L. pentosus and yeasts were the only microbial members of the biofilm formed during fermentation, regardless of inoculation. Hence, the green olive surface provides an appropriate environmental condition for the suitable development and formation of complex biofilms during controlled or natural table olive processing. PMID:22656327
Dom�nguez-Manzano, Jes�s; Le�n-Romero, �ngela; Olmo-Ruiz, Carmen; Bautista-Gallego, Joaqu�n; Arroyo-L�pez, Francisco No�; Garrido-Fern�ndez, Antonio; Jim�nez-D�az, Rufino
Heparin-binding motifs and biofilm formation by Candida albicans.
Candida albicans is a leading pathogen in infections of central venous catheters, which are frequently infused with heparin. Binding of C. albicans to medically relevant concentrations of soluble and plate-bound heparin was demonstrable by confocal microscopy and enzyme-linked immunosorbent assay (ELISA). A sequence-based search identified 34 C. albicans surface proteins containing ?1 match to linear heparin-binding motifs. The virulence factor Int1 contained the most putative heparin-binding motifs (n = 5); peptides encompassing 2 of 5 motifs bound to heparin-Sepharose. Alanine substitution of lysine residues K805/K806 in 804QKKHQIHK811 (motif 1 of Int1) markedly attenuated biofilm formation in central venous catheters in rats, whereas alanine substitution of K1595/R1596 in 1593FKKRFFKL1600 (motif 4 of Int1) did not impair biofilm formation. Affinity-purified immunoglobulin G (IgG) recognizing motif 1 abolished biofilm formation in central venous catheters; preimmune IgG had no effect. After heparin treatment of C. albicans, soluble peptides from multiple C. albicans surface proteins were detected, such as Eno1, Pgk1, Tdh3, and Ssa1/2 but not Int1, suggesting that heparin changes candidal surface structures and may modify some antigens critical for immune recognition. These studies define a new mechanism of biofilm formation for C. albicans and a novel strategy for inhibiting catheter-associated biofilms. PMID:23904295
Green, Julianne V; Orsborn, Kris I; Zhang, Minlu; Tan, Queenie K G; Greis, Kenneth D; Porollo, Alexey; Andes, David R; Long Lu, Jason; Hostetter, Margaret K
Inhibition of biofilm formation and antibacterial properties of a silver nano-coating on human dentine.
The survival of pathogenic bacteria in the oral cavity depends on their successful adhesion to dental surfaces and their ability to develop into biofilms, known as dental plaque. Bacteria from the dental plaque are responsible for the development of dental caries, gingivitis, periodontitis, stomatitis and peri-implantitis. Certain metal nanoparticles have been suggested for infection control and the management of the oral biofilm. Here, it is shown that application of a silver nano-coating directly on dentine can successfully prevent the biofilm formation on dentine surfaces as well as inhibit bacterial growth in the surrounding media. This silver nano-coating was found to be stable (>98.8%) and to maintain its integrity in biological fluids. Its antibacterial activity was compared to silver nitrate and the widely used clinical antiseptic, chlorhexidine. The bacterial growth and cell viability were quantitatively assessed by measuring the turbidity, proportion of live and dead cells and lactate production. All three bioassays showed that silver nanoparticles and silver nitrate dentine coatings were equally highly bactericidal (>99.5%), while inhibiting bacterial adhesion. However, the latter caused significant dentine discolouration (?E* = 50.3). The chlorhexidine coating showed no antibacterial effect. Thus, silver nanoparticles may be a viable alternative to both chlorhexidine and silver nitrate, protecting from dental plaque and secondary caries when applied as a dentine coating, while they may provide the platform for creating anti-biofilm surfaces in medical devices and other biomedical applications. PMID:23875717
Besinis, Alexandros; De Peralta, Tracy; Handy, Richard D
BpsR Modulates Bordetella Biofilm Formation by Negatively Regulating the Expression of the Bps Polysaccharide
Bordetella bacteria are Gram-negative respiratory pathogens of animals, birds, and humans. A hallmark feature of some Bordetella species is their ability to efficiently survive in the respiratory tract even after vaccination. Bordetella bronchiseptica and Bordetella pertussis form biofilms on abiotic surfaces and in the mouse respiratory tract. The Bps exopolysaccharide is one of the critical determinants for biofilm formation and the survival of Bordetella in the murine respiratory tract. In order to gain a better understanding of regulation of biofilm formation, we sought to study the mechanism by which Bps expression is controlled in Bordetella. Expression of bpsABCD (bpsA-D) is elevated in biofilms compared with levels in planktonically grown cells. We found that bpsA-D is expressed independently of BvgAS. Subsequently, we identified an open reading frame (ORF), BB1771 (designated here bpsR), that is located upstream of and in the opposite orientation to the bpsA-D locus. BpsR is homologous to the MarR family of transcriptional regulators. Measurement of bpsA and bpsD transcripts and the Bps polysaccharide levels from the wild-type and the ?bpsR strains suggested that BpsR functions as a repressor. Consistent with enhanced production of Bps, the bpsR mutant displayed considerably more structured biofilms. We mapped the bpsA-D promoter region and showed that purified BpsR protein specifically bound to the bpsA-D promoter. Our results provide mechanistic insights into the regulatory strategy employed by Bordetella for control of the production of the Bps polysaccharide and biofilm formation.
Conover, Matt S.; Redfern, Crystal J.; Ganguly, Tridib; Sukumar, Neelima; Sloan, Gina; Mishra, Meenu
Spatial and temporal dynamics of cellulose degradation and biofilm formation by Caldicellulosiruptor obsidiansis and Clostridium thermocellum Caldicellulosiruptor obsidiansis
Cellulose degradation is one of the major bottlenecks of a consolidated bioprocess that employs cellulolytic bacterial cells as catalysts to produce biofuels from cellulosic biomass. In this study, we investigated the spatial and temporal dynamics of cellulose degradation by Caldicellulosiruptor obsidiansis, which does not produce cellulosomes, and Clostridium thermocellum, which does produce cellulosomes. Results showed that the degradation of either regenerated or natural cellulose was synchronized with biofilm formation, a process characterized by the formation and fusion of numerous crater-like depressions on the cellulose surface. In addition, the dynamics of biofilm formation were similar in both bacteria, regardless of cellulosome production. Only the areas of cellulose surface colonized by microbes were significantly degraded, highlighting the essential role of the cellulolytic biofilm in cellulose utilization. After initial attachment, the microbial biofilm structure remained thin, uniform and dense throughout the experiment. A cellular automaton model, constructed under the assumption that the attached cells divide and produce daughter cells that contribute to the hydrolysis of the adjacent cellulose, can largely simulate the observed process of biofilm formation and cellulose degradation. This study presents a model, based on direct observation, correlating cellulolytic biofilm formation with cellulose degradation.
Wang, Zhiwu [ORNL; Lee, Sueng-Hwan [National Institute of Advanced Industrial Science and Technology, Japan; Elkins, James G [ORNL; Morrell-Falvey, Jennifer L [ORNL
The biocide chlorine dioxide stimulates biofilm formation in Bacillus subtilis by activation of the histidine kinase KinC.
Bacillus subtilis forms biofilms in response to signals that remain poorly defined. We report that biofilm formation is stimulated by sublethal doses of chlorine dioxide (ClO(2)), an extremely effective and fast-acting biocide. ClO(2) accelerated biofilm formation in B. subtilis as well as in other bacteria, suggesting that biofilm formation is a widely conserved response to sublethal doses of the agent. Biofilm formation depends on the synthesis of an extracellular matrix that holds the constituent cells together. We show that the transcription of the major operons responsible for the matrix production in B. subtilis, epsA-epsO and yqxM-sipW-tasA, was enhanced by ClO(2), in a manner that depended on the membrane-bound kinase KinC. Activation of KinC appeared to be due to the ability of ClO(2) to collapse the membrane potential. Importantly, strains unable to make a matrix were hypersensitive to ClO(2), indicating that biofilm formation is a defensive response that helps protect cells from the toxic effects of the biocide. PMID:20971918
Shemesh, Moshe; Kolter, Roberto; Losick, Richard
Probiotic-mediated competition, exclusion and displacement in biofilm formation by food-borne pathogens.
The objective of this study was to examine the inhibitory effect of probiotic strains on pathogenic biofilm formation in terms of competition, exclusion and displacement. Probiotic strains (Lactobacillus acidophilus KACC 12419, Lact.�casei KACC 12413, Lact.�paracasei KACC 12427 and Lact.�rhamnosus KACC 11953) and pathogens (Salmonella Typhimurium KCCM 40253 and Listeria monocytogenes KACC 12671) were used to evaluate the auto-aggregation, hydrophobicity and biofilm formation inhibition. The highest auto-aggregation abilities were observed in Lact.�rhamnosus (17�5%), Lact.�casei (17�2%) and Lact.�acidophilus (15�1%). Salm.�Typhimurium had the highest affinity to xylene, showing the hydrophobicity of 53�7%. The numbers of L.�monocytogenes biofilm cells during the competition, exclusion and displacement assays were effectively reduced by more than 3 log when co-cultured with Lact.�paracasei and Lact.�rhamnosus. The results suggest that probiotic strains can be used as alternative way to effectively reduce the biofilm formation in pathogenic bacteria through competition, exclusion and displacement. PMID:23362863
Woo, J; Ahn, J
Control of pathogen growth and biofilm formation using a urinary catheter that releases antimicrobial nitrogen oxides.
Antibacterial nitrogen oxides including nitric oxide are formed from nitrite under acidic conditions. In a continuous-flow model of the urinary bladder we used the retention cuff of an all-silicone Foley catheter as a depot for export of nitrogen oxides. The cuff was filled with sodium nitrite (50mM) and an acidic buffer solution (pH 3.6) and the growth of nine common uropathogens in the surrounding artificial urine was measured along with biofilm formation on the catheter surface. In experiments with control catheters (NaCl) bacteria grew readily and biofilm developed within hours in five of nine strains. In contrast, with test catheters bacterial counts were markedly reduced and biofilm formation by Pseudomonas aeruginosa, Klebsiella pneumoniae, and Enterobacter cloace was prevented, whereas Escherichia coli and Staphylococcus aureus were unaffected. We conclude that antibacterial nitrogen oxides generated in the retention cuff of a urinary catheter diffuse into urine and prevent the growth of urinary pathogens and biofilm formation. Although promising, future studies will reveal if this novel approach can be clinically useful for the prevention of catheter-associated urinary tract infections. PMID:24084579
Kishikawa, Hiroaki; Ebberyd, Anette; R�mling, Ute; Brauner, Annelie; L�thje, Petra; Lundberg, Jon O; Weitzberg, Eddie
Aggregatibacter actinomycetemcomitans arcB influences hydrophobic properties, biofilm formation and adhesion to hydroxyapatite
The regulation of gene expression in the oral pathogen Aggregatibacter actinomycetemcomitans is still not fully elucidated. ArcAB is a two-component system which allows facultative anaerobic bacteria to sense various respiratory growth conditions and adapt their gene expression accordingly.This study investigated in A. actinomycetemcomitans the role of ArcB on the regulation of biofilm formation, adhesion to saliva coated hydroxyapatite (SHA) and the hydrophobic properties of the cell. These phenotypic traits were determined for an A. actinomycetemcomitans arcB deficient type and a wild type strain. Differences in hydrophobic properties were shown at early and late exponential growth phases under microaerobic incubation and at late exponential phase under anaerobiosis.The arcB mutant formed less biofilm than the wild type strain when grown under anaerobic incubation, but displayed higher biofilm formation activity under microaerobic conditions. The adherence to SHA was significantly lower in the mutant when compared with the wild type strain. These results suggest that the transmembrane sensor kinase ArcB, in A. actinomycetemcomitans, senses redox growth conditions and regulates the expression of surface components of the bacterial cell related to biofilm formation and adhesion to saliva coated surfaces.
Longo, PL; Ota-Tsuzuki, C; Nunes, ACR; Fernandes, BL; Mintz, K; Fives-Taylor, P; Mayer, MPA
Antibacterial activity of moxifloxacin on bacteria associated with periodontitis within a biofilm.
The activity of moxifloxacin was compared with ofloxacin and doxycycline against bacteria associated with periodontitis within a biofilm (single strain and mixed population) in vitro. MICs and minimal bactericidal concentrations (MBCs) of moxifloxacin, ofloxacin and doxycyline were determined against single strains and mixed populations in a planktonic state. Single-species biofilms of two Porphyromonas gingivalis and two Aggregatibacter actinomycetemcomitans strains and a multispecies biofilm consisting of 12 species were formed for 3 days. The minimal biofilm eradication concentrations (MBECs) were determined after exposing the biofilms to the antibacterials (0.002-512 �g ml(-1)) for 18 h, addition of nutrient broth for 3 days and subsequent subcultivation. Photographs were taken using confocal laser-scanning microscopy and scanning electron microscopy. The MICs and MBCs did not differ between ofloxacin and moxifloxacin against A. actinomycetemcomitans, whilst moxifloxacin was more active than the other tested antibacterials against anaerobes and the mixed population. The single-species biofilms were eradicated by moderate concentrations of the antibacterials, and the lowest MBECs were always found for moxifloxacin (2-8 �g ml(-1)). MBECs against the multispecies biofilms were 128, >512 and >512 �g ml(-1) for moxifloxacin, ofloxacin and doxycycline, respectively. In summary, moxifloxacin in a topical formulation may have potential as an adjunct to mechanical removal of the biofilms. PMID:24217128
Tsaousoglou, Phoebus; Nietzsche, Sandor; Cachovan, Georg; Sculean, Anton; Eick, Sigrun
Characterization of biofilm formation by Riemerella anatipestifer
Riemerella anatipestifer (RA) causes epizootics of infectious disease in poultry and results in serious economic losses, especially for the duck industry. The present study focuses on understanding the biofilm-producing ability of RA strains in attempt to explain the intriguing persistence of RA post-infection on duck farms. Four RA serotype reference strains and 39 field RA isolates were measured for the
Qinghai Hu; Xiangan Han; Xiaojin Zhou; Siyu Ding; Chan Ding; Shengqing Yu
Chemically Specific Cellular Imaging of Biofilm Formation.
National Technical Information Service (NTIS)
This document and the accompanying manuscripts summarize the technical accomplishments for our one-year LDRD-ER effort. Biofilm forming microbes have existed on this planet for billions of years and make up 60% of the biological mass on earth. Such microb...
J. L. Herberg C. Schaldach J. Horn E. Gjersing R. Maxwell
Biofilm formation during the start-up period of an anaerobic biofilm reactor�Impact of nutrient complementation
This study proposes an approach to understand biofilm formation in polyphasic anaerobic digestion systems. The objective was to better control biofilm formation in order to reduce the time of colonization during the start-up phase of an anaerobic high-rate biofilm reactor.A three-phase (liquid�gas�solid) inverse turbulent bed reactor (ITBR), with low-density particles used as biomass carrier, was started by stepwise increase of
R. Cresson; H. Carr�re; J. P. Delgen�s; N. Bernet
Role of Rhizobium endoglucanase CelC2 in cellulose biosynthesis and biofilm formation on plant roots and abiotic surfaces
Background The synthesis of cellulose is among the most important but poorly understood biochemical processes, especially in bacteria, due to its complexity and high degree of regulation. In this study, we analyzed both the production of cellulose by all known members of the Rhizobiaceae and the diversity of Rhizobium celABC operon predicted to be involved in cellulose biosynthesis. We also investigated the involvement in cellulose production and biofilm formation of celC gene encoding an endoglucanase (CelC2) that is required for canonical symbiotic root hair infection by Rhizobium leguminosarum bv. trifolii. Results ANU843 celC mutants lacking (ANU843?C2) or overproducing cellulase (ANU843C2+) produced greatly increased or reduced amounts of external cellulose micro fibrils, respectively. Calcofluor-stained cellulose micro fibrils were considerably longer when formed by ANU843?C2 bacteria rather than by the wild-type strain, in correlation with a significant increase in their flocculation in batch culture. In contrast, neither calcofluor-stained extracellular micro fibrils nor flocculation was detectable in ANU843C2+ cells. To clarify the role of cellulose synthesis in Rhizobium cell aggregation and attachment, we analyzed the ability of these mutants to produce biofilms on different surfaces. Alteration of wild-type CelC2 levels resulted in a reduced ability of bacteria to form biofilms both in abiotic surfaces and in planta. Conclusions Our results support a key role of the CelC2 cellulase in cellulose biosynthesis by modulating the length of the cellulose fibrils that mediate firm adhesion among Rhizobium bacteria leading to biofilm formation. Rhizobium cellulose is an essential component of the biofilm polysaccharidic matrix architecture and either an excess or a defect of this �building material� seem to collapse the biofilm structure. These results position cellulose hydrolytic enzymes as excellent anti-biofilm candidates.
Biofilm Formation by Mycobacterium bovis: Influence of Surface Kind and Temperatures of Sanitizer Treatments on Biofilm Control
Mycobacterium bovis causes classic bovine tuberculosis, a zoonosis which is still a concern in Africa. Biofilm forming ability of two Mycobacterium bovis strains was assessed on coupons of cement, ceramic, or stainless steel in three different microbiological media at 37�C with agitation for 2, 3, or 4 weeks to determine the medium that promotes biofilm. Biofilm mass accumulated on coupons was treated with 2 sanitizers (sanitizer A (5.5?mg?L?1 active iodine) and sanitizer B (170.6?g1 alkyl dimethylbenzyl ammonium chloride, 78?g?1 didecyldimethyl ammonium chloride, 107.25?g?L?1 glutaraldehyde, 146.25?g?L?1 isopropanol, and 20?g?L?1 pine oil) at 28 and 45�C and in hot water at 85�C for 5?min. Residual biofilms on treated coupons were quantified using crystal violet binding assay. The two strains had a similar ability to form biofilms on the three surfaces. More biofilms were developed in media containing 5% liver extract. Biofilm mass increased as incubation time increased till the 3rd week. More biofilms were formed on cement than on ceramic and stainless steel surfaces. Treatment with hot water at 85�C reduced biofilm mass, however, sanitizing treatments at 45�C removed more biofilms than at 28�C. However, neither treatment completely eliminated the biofilms. The choice of processing surface and temperatures used for sanitizing treatments had an impact on biofilm formation and its removal from solid surfaces.
Adetunji, Victoria O.; Kehinde, Aderemi O.; Bolatito, Olayemi K.; Chen, Jinru
Biofilm formation by coagulase-negative staphylococci: Impact on the efficacy of antimicrobials and disinfectants commonly used on dairy farms.
Coagulase-negative staphylococci (CNS) have traditionally been considered minor mastitis pathogens and are the bacteria most frequently isolated from intramammary infection. Previously, our laboratory demonstrated that a majority of CNS isolated from Canadian milk were able to form biofilm and this was strongly and positively associated with days in milk. Biofilms offer protection against antibiotics and disinfectants, and the presence of CNS biofilms near the end of the lactation cycle could have an impact on the prevention and recurrence of CNS infections in the next lactation cycle. The objective of this study was to investigate the effect of biofilm formation on efficacy of commonly used antibiotics and disinfectants against CNS. The minimal inhibitory concentration (MIC) and minimal biofilm eradication concentration (MBEC) of several CNS isolates were determined using microdilution method and the MBEC device, respectively. Biofilm cells were more resistant to a penicillin G/novobiocin combination and to ceftiofur than their planktonic counterparts and the increase in resistance ranged from 4� to 2048�. For the disinfectants, we determined the minimum contact time required for different teat disinfectants to eradicated planktonic cells and biofilms. The chlorhexidine-based teat disinfectants eradicated planktonic cells and biofilms within 30s. For iodine-based teat disinfectants, it took 2-10� longer to eradicate the biofilms than planktonic cells. In conclusion, CNS biofilms were less susceptible to antibiotics; however, chlorhexidine-based teat disinfectants were still effective against CNS biofilms. This reinforces the use of post-milking teat disinfectants as a preventive measure of intramammary infections. PMID:24984943
Tremblay, Yannick D N; Caron, Vincent; Blondeau, Andr�anne; Messier, Serge; Jacques, Mario
Studies to control biofilm formation by coupling ultrasonication of natural waters and anodization of titanium.
The main objective of this study was to investigate the combined effect of ultrasonication of natural waters and anodization of titanium on microbial density and biofilm formation tendency on titanium surfaces. Application of 24 kHz, 400 W high power ultrasound through a 14 mm horn type SS (stainless steel) Sonicator with medium amplitude of 60% for 30 min brought about three order decrease in total bacterial density of laboratory tap water, cooling tower water and reservoir water and two order decrease in seawater. Studies on the effect of ultrasonication on dilute pure cultures of Gram-negative and Gram-positive bacteria showed five order and three order decrease for Pseudomonas sp. and Flavobacterium sp. respectively and two order and less than one order decrease for Bacillus sp. and Micrococcus sp. respectively. Ultrasonication increased lag phase and reduced logarithmic population increase and specific growth rate of Gram-negative bacteria whereas for Gram-positive bacteria specific growth rate increased. Studies on the biofilm formation tendency of these ultrasonicated mediums on titanium surface showed one order reduction under all conditions. Detailed biofilm imaging by advanced microscopic techniques like AFM, SEM and epifluorescence microscopy clearly visualized the lysed/damaged cells and membrane perforations due to ultrasonication. Combination of ultrasonication and anodization brought about maximum decrease in bacterial density and biofilm formation with greater than two order decrease in seawater, two order decrease in Bacillus sp. culture and more than four order decrease in Flavobacterium sp. culture establishing the synergistic effect of anodization and ultrasonication in this study. PMID:23871547
Nithila, S D Ruth; Anandkumar, B; Vanithakumari, S C; George, R P; Mudali, U Kamachi; Dayal, R K
Purpurin Suppresses Candida albicans Biofilm Formation and Hyphal Development
A striking and clinically relevant virulence trait of the human fungal pathogen Candida albicans is its ability to grow and switch reversibly among different morphological forms. Inhibition of yeast-to-hypha transition in C. albicans represents a new paradigm for antifungal intervention. We have previously demonstrated the novel antifungal activity of purpurin against Candida fungi. In this study, we extended our investigation by examining the in vitro effect of purpurin on C. albicans morphogenesis and biofilms. The susceptibility of C. albicans biofilms to purpurin was examined quantitatively by 2,3-bis(2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide reduction assay. Hyphal formation and biofilm ultrastructure were examined qualitatively by scanning electron microscopy (SEM). Quantitative reverse transcription-PCR (qRT-PCR) was used to evaluate the expression of hypha-specific genes and hyphal regulator in purpurin-treated fungal cells. The results showed that, at sub-lethal concentration (3 �g/ml), purpurin blocked the yeast-to-hypha transition under hypha-inducing conditions. Purpurin also inhibited C. albicans biofilm formation and reduced the metabolic activity of mature biofilms in a concentration-dependent manner. SEM images showed that purpurin-treated C. albicans biofilms were scanty and exclusively consisted of aggregates of blastospores. qRT-PCR analyses indicated that purpurin downregulated the expression of hypha-specific genes (ALS3, ECE1, HWP1, HYR1) and the hyphal regulator RAS1. The data strongly suggested that purpurin suppressed C. albicans morphogenesis and caused distorted biofilm formation. By virtue of the ability to block these two virulence traits in C. albicans, purpurin may represent a potential candidate that deserves further investigations in the development of antifungal strategies against this notorious human fungal pathogen in vivo.
Tsang, Paul Wai-Kei; Bandara, H. M. H. N.; Fong, Wing-Ping
Biofilm formation by oral clinical isolates of Candida species.
We have conducted a longitudinal study to quantify biofilms in oral clinical isolates of Candida species (spp.) from adults with local and systemic predisposing factors for candidiasis. A total of 69 yeast isolates from 63 Mexican patients were evaluated. These isolates (39 C. albicans, 15 C. tropicalis, 7 C. glabrata, 4 C. krusei, 1 C. lusitaniae, 1 C. kefyr, 1 C. guilliermondii and 1 C. pulcherrima) were obtained from two clinical sites: 62.3% (n=43) from the oral mucosa of totally and partially edentulous patients, and 37.7% (n=26) from the oral mucosa of diabetics. In addition, Candida ATCC strains were used as controls for each experiment. The kinetics of biofilm formation were measured by 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide [XTT] reduction; each isolate was tested at 6, 12 and 24h. Biofilm formation is dependent on the Candida spp. and its clinical origin. On average, the oral isolates of C. glabrata are strong biofilm producers, whereas C. albicans and C. tropicalis are moderate producers. The most common species in our population was C. albicans. While the kinetics of C. albicans biofilm formation varies between oral isolates, it generally maintains steady growth from 2 to 48h, when it reaches its maximum growth. PMID:23849353
S�nchez-Vargas, Luis Octavio; Estrada-Barraza, Deyanira; Pozos-Guillen, Amaury J; Rivas-Caceres, Raimundo
Effect of cyclic bis(3?-5?)diguanylic acid and its analogs on bacterial biofilm formation
Cyclic bis(3?�5?)diguanylic acid (cyclic-di-GMP) functions as a second messenger in diverse species of bacteria to trigger wide-ranging physiological changes. We measured cyclic-di-GMP and its structural analogs such as cyclic bis(3?�5?)guanylic/adenylic acid (cyclic-GpAp), cyclic bis(3?�5?)guanylic/inosinic acid (cyclic-GpIp) and monophosphorothioic acid of cyclic-di-GMP (cyclic-GpGps) for effects on the biofilm formation of Staphylococcus aureus and Pseudomonas aeruginosa. We constructed a knockout mutant of SA0701, which is a GGDEF motif protein relevant to diguanylate cyclase from S. aureus 2507. We confirmed that the biofilm formation of this mutant (MS2507?SA0701) was reduced. Cyclic-di-GMP corresponding to physiological intracellular levels given in the culture recovered the biofilm formation of MS2507?SA0701, whereas its analogs did not, indicating that unlike a previous suggestion, cyclic-di-GMP was involved in the positive regulation of the biofilm formation of S. aureus and its action was structurally specific. At a high concentration (200 ?M), cyclic-di-GMP and its analogs showed suppression effects on the biofilm formation of S. aureus and P. aeruginosa, and according to the quantification study using costat analysis, the suppression potential was in the order of cyclic-di-GMP, cyclic-GpGps, cyclic-GpAp and cyclic-GpIp, suggesting that the suppression effect was not strictly specific and the change of base structure quantitatively affected the suppression activity.
Ishihara, Yuka; Hyodo, Mamoru; Hayakawa, Yoshihiro; Kamegaya, Taichi; Yamada, Keiko; Okamoto, Akira; Hasegawa, Tadao; Ohta, Michio
Systematic analysis of diguanylate cyclases that promote biofilm formation by Pseudomonas fluorescens Pf0-1.
Cyclic di-GMP (c-di-GMP) is a broadly conserved, intracellular second-messenger molecule that regulates biofilm formation by many bacteria. The synthesis of c-di-GMP is catalyzed by diguanylate cyclases (DGCs) containing the GGDEF domain, while its degradation is achieved through the phosphodiesterase activities of EAL and HD-GYP domains. c-di-GMP controls biofilm formation by Pseudomonas fluorescens Pf0-1 by promoting the cell surface localization of a large adhesive protein, LapA. LapA localization is regulated posttranslationally by a c-di-GMP effector system consisting of LapD and LapG, which senses cytoplasmic c-di-GMP and modifies the LapA protein in the outer membrane. Despite the apparent requirement for c-di-GMP for biofilm formation by P. fluorescens Pf0-1, no DGCs from this strain have been characterized to date. In this study, we undertook a systematic mutagenesis of 30 predicted DGCs and found that mutations in just 4 cause reductions in biofilm formation by P. fluorescens Pf0-1 under the conditions tested. These DGCs were characterized genetically and biochemically to corroborate the hypothesis that they function to produce c-di-GMP in vivo. The effects of DGC gene mutations on phenotypes associated with biofilm formation were analyzed. One DGC preferentially affects LapA localization, another DGC mainly controls swimming motility, while a third DGC affects both LapA and motility. Our data support the conclusion that different c-di-GMP-regulated outputs can be specifically controlled by distinct DGCs. PMID:21764921
Newell, Peter D; Yoshioka, Shiro; Hvorecny, Kelli L; Monds, Russell D; O'Toole, George A
Atomic force microscopy of bacteria from periodontal subgingival biofilm: Preliminary study results
Objective: Atomic force microscope (AFM) is a technology that allows analysis of the nanoscale morphology of bacteria within biofilm and provides details that may be better useful for understanding the role of bacterial interactions in the periodontal disease. Material and Methods: Five patients with periodontal ?5 mm pockets diagnosed as generalized periodontitis and five patients with slight gingivitis were selected for the investigation. Bacteria biofilms were collected and morphologically investigated by AFM application. Results: The investigation revealed how periodontitis bacteria are characterized by specific morphologic features of the cell wall. The major representative species of bacteria causing periodontal diseases have been reproduced by a three-dimensional reconstruction showing the bacteria surface details. Conclusions: The presence of complex glycocalyx structures, bacteriophage-like vesicles, spirochetes (classic and cystic morphology) and bacterial co-aggregation has been identified by the AFM analysis. The results suggest that AFM is a reliable technique for studying bacterial morphology and for examining microbial interactions in dental plaque.
Germano, Francesco; Bramanti, Ennio; Arcuri, Claudio; Cecchetti, Francesco; Cicciu, Marco
Bacteriophage Cocktail for the Prevention of Biofilm Formation by Pseudomonas aeruginosa on Catheters in an In Vitro Model System?
Microorganisms develop biofilms on indwelling medical devices and are associated with device-related infections, resulting in substantial morbidity and mortality. This study investigated the effect of pretreating hydrogel-coated catheters with Pseudomonas aeruginosa bacteriophages on biofilm formation by P. aeruginosa in an in vitro model. Hydrogel-coated catheters were exposed to a 10 log10 PFU ml?1 lysate of P. aeruginosa phage M4 for 2 h at 37�C prior to bacterial inoculation. The mean viable biofilm count on untreated catheters was 6.87 log10 CFU cm?2 after 24 h. The pretreatment of catheters with phage reduced this value to 4.03 log10 CFU cm?2 (P < 0.001). Phage treatment immediately following bacterial inoculation also reduced biofilm viable counts (4.37 log10 CFU cm?2 reduction; P < 0.001). The regrowth of biofilms on phage-treated catheters occurred between 24 and 48 h, but supplemental treatment with phage at 24 h significantly reduced biofilm regrowth (P < 0.001). Biofilm isolates resistant to phage M4 were recovered from catheters pretreated with phage. The phage susceptibility profiles of these isolates were used to guide the development of a five-phage cocktail from a larger library of P. aeruginosa phages. The pretreatment of catheters with this cocktail reduced the 48-h mean biofilm cell density by 99.9% (from 7.13 to 4.13 log10 CFU cm?2; P < 0.001), but fewer biofilm isolates were resistant to these phages. These results suggest the potential of applying phages, especially phage cocktails, to the surfaces of indwelling medical devices for mitigating biofilm formation by clinically relevant bacteria.
Fu, Weiling; Forster, Terri; Mayer, Oren; Curtin, John J.; Lehman, Susan M.; Donlan, Rodney M.
Biofilm formation, host-cell adherence, and virulence genes regulation of Streptococcus suis in response to autoinducer-2 signaling.
Autoinducer-2 (AI-2) is a universal signal molecule mediating intra- and interspecies communication among bacteria. AI-2 is a byproduct of the LuxS enzyme during the catabolism of S-adenosylhomocysteine and plays critical roles in regulating various behaviors of bacteria. In our previous study, the function of LuxS in AI-2 production was verified in Streptococcus suis (SS). Decreased levels of SS biofilm formation and host-cell adherence as well as the inability to produce AI-2 were observed in SS having a luxS mutant gene. In this study, exogenous addition of a low concentration of AI-2 synthesized in vitro was found to promote biofilm formation and host-cell adherence. However, higher concentrations of AI-2 inhibited SS biofilm formation and host-cell adherence. Real-time PCR results showed that the mRNA level of virulence factors of SS biofilm, gdh, cps2, sly, and mrp increased and ef, fbps, and gapdh decreased with increasing AI-2 concentrations. These findings demonstrated that AI-2 supplemented exogenously acted as a concentration-dependent signaling molecule to regulate SS biofilm formation, host-cell adherence, and transcription levels of many virulence genes. PMID:24370626
Wang, Yang; Yi, Li; Zhang, Zhicheng; Fan, Hongjie; Cheng, Xiangchao; Lu, Chengping
Inhibitory effects of lactoferrin on growth and biofilm formation of Porphyromonas gingivalis and Prevotella intermedia.
Lactoferrin (LF) is an iron-binding antimicrobial protein present in saliva and gingival crevicular fluids, and it is possibly associated with host defense against oral pathogens, including periodontopathic bacteria. In the present study, we evaluated the in vitro effects of LF-related agents on the growth and biofilm formation of two periodontopathic bacteria, Porphyromonas gingivalis and Prevotella intermedia, which reside as biofilms in the subgingival plaque. The planktonic growth of P. gingivalis and P. intermedia was suppressed for up to 5 h by incubation with >or=130 microg/ml of human LF (hLF), iron-free and iron-saturated bovine LF (apo-bLF and holo-bLF, respectively), and >or=6 microg/ml of bLF-derived antimicrobial peptide lactoferricin B (LFcin B); but those effects were weak after 8 h. The biofilm formation of P. gingivalis and P. intermedia over 24 h was effectively inhibited by lower concentrations (>or=8 microg/ml) of various iron-bound forms (the apo, native, and holo forms) of bLF and hLF but not LFcin B. A preformed biofilm of P. gingivalis and P. intermedia was also reduced by incubation with various iron-bound bLFs, hLF, and LFcin B for 5 h. In an examination of the effectiveness of native bLF when it was used in combination with four antibiotics, it was found that treatment with ciprofloxacin, clarithromycin, and minocycline in combination with native bLF for 24 h reduced the amount of a preformed biofilm of P. gingivalis compared with the level of reduction achieved with each agent alone. These results demonstrate the antibiofilm activity of LF with lower iron dependency against P. gingivalis and P. intermedia and the potential usefulness of LF for the prevention and treatment of periodontal diseases and as adjunct therapy for periodontal diseases. PMID:19451301
Wakabayashi, Hiroyuki; Yamauchi, Koji; Kobayashi, Tetsuo; Yaeshima, Tomoko; Iwatsuki, Keiji; Yoshie, Hiromasa
EmbRS a new two-component system that inhibits biofilm formation and saves Rubrivivax gelatinosus from sinking
Photosynthetic bacteria can switch from planktonic lifestyle to phototrophic biofilm in mats in response to environmental changes. The mechanisms of phototrophic biofilm formation are, however, not characterized. Herein, we report a two-component system EmbRS that controls the biofilm formation in a photosynthetic member of the Burkholderiales order, the purple bacterium Rubrivivax gelatinosus. EmbRS inactivation results in cells that form conspicuous bacterial veils and fast-sinking aggregates in liquid. Biofilm analyses indicated that EmbRS represses the production of an extracellular matrix and biofilm formation. Mapping of transposon mutants that partially or completely restore the wild-type (WT) phenotype allowed the identification of two gene clusters involved in polysaccharide synthesis, one fully conserved only in Thauera sp., a floc-forming wastewater bacterium. A second two-component system BmfRS and a putative diguanylate cyclase BdcA were also identified in this screen suggesting their involvement in biofilm formation in this bacterium. The role of polysaccharides in sinking of microorganisms and organic matter, as well as the importance and the evolution of such regulatory system in phototrophic microorganisms are discussed.
Steunou, Anne Soisig; Liotenberg, Sylviane; Soler, Marie-Noelle; Briandet, Romain; Barbe, Valerie; Astier, Chantal; Ouchane, Soufian
Colorimetric Method for Identifying Plant Essential Oil Components That Affect Biofilm Formation and Structure
The specific biofilm formation (SBF) assay, a technique based on crystal violet staining, was developed to locate plant essential oils and their components that affect biofilm formation. SBF analysis determined that cinnamon, cassia, and citronella oils differentially affected growth-normalized biofilm formation by Escherichia coli. Examination of the corresponding essential oil principal components by the SBF assay revealed that cinnamaldehyde decreased
C. Niu; E. S. Gilbert
Production of Antibacterial Compounds and Biofilm Formation by Roseobacter Species Are Influenced by Culture Conditions? �
Bacterial communities associated with marine algae are often dominated by members of the Roseobacter clade, and in the present study, we describe Roseobacter phenotypes that may provide this group of bacteria with selective advantages when colonizing this niche. Nine of 14 members of the Roseobacter clade, of which half were isolated from cultures of the dinoflagellate Pfiesteria piscicida, produced antibacterial compounds. Many non-Roseobacter marine bacteria were inhibited by sterile filtered supernatants of Silicibacter sp. TM1040 and Phaeobacter (formerly Roseobacter) strain 27-4, which had the highest production of antibacterial compound. In contrast, Roseobacter strains were susceptible only when exposed to concentrated compound. The production of antibacterial compound was influenced by the growth conditions, as production was most pronounced when bacteria were grown in liquid medium under static conditions. Under these conditions, Silicibacter sp. TM1040 cells attached to one another, forming rosettes, as has previously been reported for Phaeobacter 27-4. A spontaneous Phaeobacter 27-4 mutant unable to form rosettes was also defective in biofilm formation and the production of antibacterial compound, indicating a possible link between these phenotypes. Rosette formation was observed in 8 of 14 Roseobacter clade strains examined and was very pronounced under static growth in 5 of these strains. Attachment to surfaces and biofilm formation at the air-liquid interface by these five strains was greatly facilitated by growth conditions that favored rosette formation, and rosette-forming strains were 13 to 30 times more efficient in attaching to glass compared to strains under conditions where rosette formation was not pronounced. We hypothesize that the ability to produce antibacterial compounds that principally inhibit non-Roseobacter species, combined with an enhancement in biofilm formation, may give members of the Roseobacter clade a selective advantage and help to explain the dominance of members of this clade in association with marine algal microbiota.
Bruhn, Jesper Bartholin; Gram, Lone; Belas, Robert
Retrocyclin inhibits Gardnerella vaginalis biofilm formation and toxin activity
Background Retrocyclins are cyclic antimicrobial peptides that have been shown to be both broadly active and safe in animal models. RC-101, a synthetic retrocyclin, targets important human pathogens and is a candidate vaginal microbicide. Its activity against microbes associated with bacterial vaginosis is unknown. Methods We investigated the effect of RC-101 on toxin activity, bacterial growth and biofilm formation of Gardnerella vaginalis in vitro. Results RC-101 potently inhibits the cytolytic activity of vaginolysin, the Gardnerella vaginalis toxin, on both erythrocytes and nucleated cells. RC-101 lacks inhibitory activity against planktonic G. vaginalis but markedly decreases biofilm formation. Conclusions These dual properties, toxin inhibition and biofilm retardation, justify further exploration of RC-101 as a candidate agent for bacterial vaginosis prevention.
Hooven, Thomas A.; Randis, Tara M.; Hymes, Saul R.; Rampersaud, Ryan; Ratner, Adam J.
Electroactivity of phototrophic river biofilms and constitutive cultivable bacteria.
Electroactivity is a property of microorganisms assembled in biofilms that has been highlighted in a variety of environments. This characteristic was assessed for phototrophic river biofilms at the community scale and at the bacterial population scale. At the community scale, electroactivity was evaluated on stainless steel and copper alloy coupons used both as biofilm colonization supports and as working electrodes. At the population scale, the ability of environmental bacterial strains to catalyze oxygen reduction was assessed by cyclic voltammetry. Our data demonstrate that phototrophic river biofilm development on the electrodes, measured by dry mass and chlorophyll a content, resulted in significant increases of the recorded potentials, with potentials of up to +120 mV/saturated calomel electrode (SCE) on stainless steel electrodes and +60 mV/SCE on copper electrodes. Thirty-two bacterial strains isolated from natural phototrophic river biofilms were tested by cyclic voltammetry. Twenty-five were able to catalyze oxygen reduction, with shifts of potential ranging from 0.06 to 0.23 V, cathodic peak potentials ranging from -0.36 to -0.76 V/SCE, and peak amplitudes ranging from -9.5 to -19.4 ?A. These isolates were diversified phylogenetically (Actinobacteria, Firmicutes, Bacteroidetes, and Alpha-, Beta-, and Gammaproteobacteria) and exhibited various phenotypic properties (Gram stain, oxidase, and catalase characteristics). These data suggest that phototrophic river biofilm communities and/or most of their constitutive bacterial populations present the ability to promote electronic exchange with a metallic electrode, supporting the following possibilities: (i) development of electrochemistry-based sensors allowing in situ phototrophic river biofilm detection and (ii) production of microbial fuel cell inocula under oligotrophic conditions. PMID:21642402
Lyautey, Emilie; Cournet, Amandine; Morin, Soizic; Boul�treau, St�phanie; Etcheverry, Luc; Charcosset, Jean-Yves; Delmas, Fran�ois; Bergel, Alain; Garabetian, Fr�d�ric
Electroactivity of Phototrophic River Biofilms and Constitutive Cultivable Bacteria ? �
Electroactivity is a property of microorganisms assembled in biofilms that has been highlighted in a variety of environments. This characteristic was assessed for phototrophic river biofilms at the community scale and at the bacterial population scale. At the community scale, electroactivity was evaluated on stainless steel and copper alloy coupons used both as biofilm colonization supports and as working electrodes. At the population scale, the ability of environmental bacterial strains to catalyze oxygen reduction was assessed by cyclic voltammetry. Our data demonstrate that phototrophic river biofilm development on the electrodes, measured by dry mass and chlorophyll a content, resulted in significant increases of the recorded potentials, with potentials of up to +120 mV/saturated calomel electrode (SCE) on stainless steel electrodes and +60 mV/SCE on copper electrodes. Thirty-two bacterial strains isolated from natural phototrophic river biofilms were tested by cyclic voltammetry. Twenty-five were able to catalyze oxygen reduction, with shifts of potential ranging from 0.06 to 0.23 V, cathodic peak potentials ranging from ?0.36 to ?0.76 V/SCE, and peak amplitudes ranging from ?9.5 to ?19.4 ?A. These isolates were diversified phylogenetically (Actinobacteria, Firmicutes, Bacteroidetes, and Alpha-, Beta-, and Gammaproteobacteria) and exhibited various phenotypic properties (Gram stain, oxidase, and catalase characteristics). These data suggest that phototrophic river biofilm communities and/or most of their constitutive bacterial populations present the ability to promote electronic exchange with a metallic electrode, supporting the following possibilities: (i) development of electrochemistry-based sensors allowing in situ phototrophic river biofilm detection and (ii) production of microbial fuel cell inocula under oligotrophic conditions.
Lyautey, Emilie; Cournet, Amandine; Morin, Soizic; Bouletreau, Stephanie; Etcheverry, Luc; Charcosset, Jean-Yves; Delmas, Francois; Bergel, Alain; Garabetian, Frederic
Impact of TiO2 Nanoparticles on Growth, Biofilm Formation, and Flavin Secretion in Shewanella oneidensis
Understanding of nanoparticle impacts on critical bacteria functions allows us to gain a mechanistic understanding of toxicity and guides us towards design rules for creating safe nanomaterials. Herein and using analytical techniques, biofilm formation, a general bacteria function, and riboflavin secretion, a species-specific function, were monitored in Shewanella oneidensis, a metal reducing bacterium, following exposure to a variety of TiO2 nanoparticle types (synthesized, Aeroxide P25, and T-Eco). TEM images show that dosed nanoparticles are in close proximity to the bacteria but they are not internalized. Using quartz crystal microbalance (QCM), it was revealed that S. oneidensis biofilm formation is slowed in the presence of nanoparticles. Though S. oneidensis grows more slowly in the presence of TiO2 nanoparticles, riboflavin secretion, a function related to the S. oneidensis metal reducing capacity, was increased significantly in a nanoparticle dose-dependent manner. Both changes in biofilm formation and riboflavin secretion are supported by changes in gene expression in nanoparticle-exposed S. oneidensis. This broad study of bacterial nanotoxicity, including use of sensitive analytical tools for functional assessments of biofilm formation, riboflavin secretion, and gene expression has implications for total ecosystem health as the use of engineered nanoparticles grows.
Maurer-Jones, Melissa A.; Gunsolus, Ian L.; Meyer, Ben M.; Christenson, Cole J.; Haynes, Christy L.
Capillary Isoelectric Focusing � Useful tool for detection of the biofilm formation in Staphylococcus epidermidis
The biofilm formation is an important factor of S. epidermidis virulence. Biofilm-positive strains might be clinically more important than biofilm-negative ones. Unlike biofilm-negative staphylococci, biofilm-positive staphylococci are surrounded with an extracellular polysaccharide substance. The presence of this substance on the surface can affect physico-chemical properties of the bacterial cell, including surface charge. 73 S. epidermidis strains were examined for the
Filip Ruzicka; Marie Horka; Veronika Hola; Miroslav Votava
Diversity of bacteria and mycobacteria in biofilms of two urban drinking water distribution systems.
In this study, to give insight into the bacterial diversity of biofilms from full-scale drinking water distribution systems (DWDSs), the bacterial community compositions of biofilms from two urban DWDSs (Guangzhou and Beijing, China) were determined using a 16S rRNA gene library technique. Meanwhile, the occurrence and diversity of mycobacteria were also analyzed by a Mycobacterium -specific hsp gene assay. The biofilms from the full-scale DWDSs have complex bacterial populations. Proteobacteria was the common and predominant group in all biofilm samples, in agreement with previous reports. The community structures of bacteria at the three sites in Guangzhou DWDS were significantly different, despite the similar physicochemical properties of portable water. Some abundant and peculiar bacterial phylotypes were noteworthy, including Methylophilus , Massilia, and Planomicrobium , members of which are rarely found in DWDSs and their roles in DWDS biofilms are still unclear. The diversity of Mycobacterium species in biofilm samples was rather low. Mycobacterium arupense and Mycobacterium gordonae were the primary Mycobacterium species in Guangzhou and Beijing biofilms, respectively, indicating that M. arupense may be more resistant to chloride than M. gordonae. PMID:22356469
Liu, Ruyin; Yu, Zhisheng; Zhang, Hongxun; Yang, Min; Shi, Baoyou; Liu, Xinchun
SarA Positively Controls Bap-Dependent Biofilm Formation in Staphylococcus aureus
The biofilm-associated protein Bap is a staphylococcal surface protein involved in biofilm formation. We investigated the influence of the global regulatory locus sarA on bap expression and Bap-dependent biofilm formation in three unrelated Staphylococcus aureus strains. The results showed that Bap-dependent biofilm formation was diminished in the sarA mutants by an agr-independent mechanism. Complementation studies using a sarA clone confirmed
M. P. Trotonda; A. C. Manna; A. L. Cheung; I. Lasa; J. R. Penades
Biofilm formation in clinical Candida isolates and its association with virulence
Biofilm formation, an important virulence trait of Candida species was measured in 107 Candida isolates from 32 candidemic patients by XTT [2,3-bis (2-methoxy-4nitro-5-sulfo-phenyl)-2H-tetra-zolium-5-carboxanilide] activity and compared to biofilm formation of Candida isolates from oropharyngeal lesions of 19 AIDS patients. Biofilm formation by XTT varied among species and C. albicans; C. lusitaniae and C. krusei produced more biofilm than the other
Fahmi Hasan; Immaculata Xess; Xiabo Wang; Neena Jain; Bettina C. Fries
Electroactive mixed culture biofilms in microbial bioelectrochemical systems: the role of temperature for biofilm formation and performance.
In this paper we investigate the temperature dependence and temperature limits of waste water derived anodic microbial biofilms. We demonstrate that these biofilms are active in a temperature range between 5�C and 45�C. Elevated temperatures during initial biofilm growth not only accelerate the biofilm formation process, they also influence the bioelectrocatalytic performance of these biofilms when measured at identical operation temperatures. For example, the time required for biofilm formation decreases from above 40 days at 15�C to 3.5 days at 35�C. Biofilms grown at elevated temperatures are more electrochemically active at these temperatures than those grown at lower incubation temperature. Thus, at 30�C current densities of 520 ?A cm(-2) and 881 ?A cm(-2) are achieved by biofilms grown at 22�C and 35�C, respectively. Vice versa, and of great practical relevance for waste water treatment plants in areas of moderate climate, at low operation temperatures, biofilms grown at lower temperatures outperform those grown at higher temperatures. We further demonstrate that all biofilms possess similar lower (0�C) and upper (50�C) temperature limits--defining the operational limits of a respective microbial fuel cell or microbial biosensor--as well as similar electrochemical electron transfer characteristics. PMID:20630740
Patil, Sunil A; Harnisch, Falk; Kapadnis, Balasaheb; Schr�der, Uwe
Molecular basis of in-vivo biofilm formation by bacterial pathogens
Summary Bacterial biofilms are involved in a multitude of serious chronic infections. In recent years, modeling biofilm infection in vitro led to the identification of microbial determinants governing biofilm development. However, we lack information as to whether biofilm formation mechanisms identified in vitro have relevance for biofilm-associated infection. Here, we discuss the molecular basis of biofilm formation using staphylococci and Pseudomonas aeruginosa to illustrate key points, as their biofilm development process is well-studied. We will focus on in-vivo findings such as obtained in animal infection models, and critically evaluate in-vivo relevance of in-vitro findings. Although results on the role of quorum-sensing in biofilm formation have been conflicting, we now argue that integration of in-vitro and in-vivo studies allows a differentiated view of this mechanism as it relates to biofilm infection.
Joo, Hwang-Soo; Otto, Michael
Although biofilm formation is widely documented on Earth, it has not been demonstrated in the absence of gravity. To explore this possibility, Pseudomonas aeruginosa, suspended in sterile buffer, was flown in a commercial payload on space shuttle flight STS-95. During earth or...
Spatial & Temporal Geophysical Monitoring of Microbial Growth and Biofilm Formation
Previous studies have examined the effect of biogenic gases and biomineralization on the acoustic properties of porous media. In this study, we investigated the spatiotemporal effect of microbial growth and biofilm formation on compressional waves and complex conductivity in sand columns. A control column (non-biostimulated) and a biostimulated column were studied in a 2D acoustic scanning apparatus, and a second
C. A. Davis; L. J. Pyrak-Nolte; E. A. Atekwana; D. D. Werkema; M. E. Haugen
Biofilm Formation by Campylobacter jejuni Is Increased under Aerobic Conditions
The microaerophilic human pathogen Campylobacter jejuni is the leading cause of food-borne bacterial gastroenteritis in the developed world. During transmission through the food chain and the environment, the organism must survive stressful environmental conditions, particularly high oxygen levels. Biofilm formation has been suggested to play a role in the environmental survival of this organism. In this work we show that
Mark Reuter; Arthur Mallett; Bruce M. Pearson; Arnoud H. M. van Vliet
Effect of Honey on Streptococcus mutans Growth and Biofilm Formation
Because of the tradition of using honey as an antimicrobial medicament, we investigated the effect of natural honey (NH) on Streptococcus mutans growth, viability, and biofilm formation compared to that of an artificial honey (AH). AH contained the sugars at the concentrations reported for NH. NH and AH concentrations were obtained by serial dilution with tryptic soy broth (TSB). Several concentrations of NH and AH were tested for inhibition of bacterial growth, viability, and biofilm formation after inoculation with S. mutans UA159 in 96-well microtiter plates to obtain absorbance and CFU values. Overall, NH supported significantly less (P < 0.05) bacterial growth than AH at 25 and 12.5% concentrations. At 50 and 25% concentrations, both honey groups provided significantly less bacterial growth and biofilm formation than the TSB control. For bacterial viability, the results for all honey concentrations except 50% NH were not significantly different from those for the TSB control. NH was able to decrease the maximum velocity of S. mutans growth compared to AH. In summary, NH demonstrated more inhibition of bacterial growth, viability, and biofilm formation than AH. This study highlights the potential antibacterial properties of NH and could suggest that the antimicrobial mechanism of NH is not solely due to its high sugar content.
Li, Mingyun
Effects of Pipe Materials on Chlorine-resistant Biofilm Formation Under Long-term High Chlorine Level.
Drinking water distribution systems are composed of various pipe materials and may harbor biofilms even in the continuous presence of disinfectants. Biofilms formation on five pipe materials (copper (Cu), polyethylene (PE), stainless steel (STS), cast iron (CI), and concrete-coated polycarbonate (CP)) within drinking water containing 1.20�mg/L free chlorine, was investigated by flow cytometry, heterotrophic plate counts, and denaturing gradient gel electrophoresis analysis. Results showed that the biofilms formation varied in pipe materials. The biofilm formed on CP initially emerged the highest biomass in 12�days, but CI presented the significantly highest biomass after 28�days, and Cu showed the lowest bacterial numbers before 120�days, while STS expressed the lowest bacterial numbers after 159�days. In the biofilm community structure, Moraxella osloensis and Sphingomonas sp. were observed in all the pipe materials while Bacillus sp. was detected except in the CP pipe and Stenotrophomonas maltophila was found from three pipe materials (Cu, PE, and STS). Other bacteria were only found from one or two pipe materials. It is noteworthy that there are 11 opportunistic pathogens in the 17 classified bacterial strains. This research has afforded crucial information regarding the influence of pipe materials on chlorine-resistant biofilm formation. PMID:24828580
Zhu, Zebing; Wu, Chenguang; Zhong, Dan; Yuan, Yixing; Shan, Lili; Zhang, Jie
Biofilm formation and antibiotic production in Ruegeria mobilis are influenced by intracellular concentrations of cyclic dimeric guanosinmonophosphate.
In many species of the marine Roseobacter clade, periods of attached life, in association with phytoplankton or particles, are interspersed with planktonic phases. The purpose of this study was to determine whether shifts between motile and sessile life in the globally abundant Roseobacter clade species Ruegeria mobilis are associated with intracellular concentrations of the signal compound cyclic dimeric guanosinmonophosphate (c-di-GMP), which in bacteria regulates transitions between motile and sessile life stages. Genes for diguanylate cyclases and phosphodiesterases, which are involved in c-di-GMP signalling, were found in the genome of R.?mobilis strain F1926. Ion pair chromatography-tandem mass spectrometry revealed 20-fold higher c-di-GMP concentrations per cell in biofilm-containing cultures than in planktonic cells. An introduced diguanylate cyclase gene increased c-di-GMP and enhanced biofilm formation and production of the potent antibiotic tropodithietic acid (TDA). An introduced phosphodiesterase gene decreased c-di-GMP and reduced biofilm formation and TDA production. tdaC, a key gene for TDA biosynthesis, was expressed only in attached or biofilm-forming cells, and expression was induced immediately after initial attachment. In conclusion, c-di-GMP signalling controls biofilm formation and biofilm-associated traits in R.?mobilis and, as suggested by presence of GGDEF and EAL domain protein genes, also in other Roseobacter clade species. PMID:24118907
D'Alvise, Paul W; Magdenoska, Olivera; Melchiorsen, Jette; Nielsen, Kristian F; Gram, Lone
Quorum quenching activity in cell-free lysate of endophytic bacteria isolated from Pterocarpus santalinus Linn., and its effect on quorum sensing regulated biofilm in Pseudomonas aeruginosa PAO1.
Quorum sensing mechanism allows the microorganisms to resist the antibiotic treatment by forming biofilms. Quorum quenching is one of the mechanisms to control the development of drug resistance in microbes. Endophyte bacteria are beneficial to plant growth as they support the immune system against the pathogen attack. The endophytic bacteria present in Pterocarpus santalinus were screened for the presence of N-acyl homoserine lactones (AHLs) degrading bacteria using biosensor strains and further confirmed by quantifying the violacein production. Cell-free lysate of endophytic bacteria, Bacillus firmus PT18 and Enterobacter asburiae PT39 exhibited potent AHL degrading ability by inhibiting about 80% violacein production in biosensor strain. Furthermore, when the cell-free lysate was applied to Pseudomonas aeruginosa PAO1 and PAO1-JP2 biofilm it resulted in significant (p<0.01) inhibition of biofilm formation. The biofilm inhibition was confirmed by visualization of biofilm slides under fluorescence microscopy, which showed decrease in total biomass formation in treated slides. Isolation and amplification of the gene (aiiA) indicated that the presence of AHL lactonase in cell-free lysate and sequence alignment indicated that AiiA contains a "HXHXDH" zinc-binding motif that is being conserved in several groups of metallohydrolases. Therefore, the study shows the potential of AHLs degradation by AHL lactonase present in cell-free lysate of isolated endophytic bacteria and inhibition of quorum sensing regulated biofilm formation in P. aeruginosa PAO1. PMID:24268182
Rajesh, P S; Ravishankar Rai, V
Effect of CaCO? particles and suspended bacteria on biofilm components and activity in the model recirculating cooling water system.
Biofilms are a serious problem in industrial recirculating cooling water systems. Biofilm formation and properties are affected by many factors, such as inorganic particles and suspended bacteria. In this research a laboratory model recirculating cooling water system was applied to investigate the effects of CaCO(3) concentration and suspended bacterial count on extracellular polymeric substances (EPS) content and dehydrogenase activity (DHA) in the attached biofilms. In addition, nutrient level was also the key factor when investigating the effect of suspended bacterial count. The results showed that EPS content and DHA first increased and then decreased with the increase of CaCO(3) concentration from 0 to 200 mg/l. At the low nutrient level, with the increase of suspended bacterial count from 4.04 to 5.78 log(10) c.f.u./ml, biofilm EPS content decreased firstly and then increased. However, biofilm DHA always gradually increased. At the medium nutrient level, biofilm EPS content increased firstly and then decreased and DHA always gradually decreased when suspended bacterial count ranged from 4.04 to 5.78 log(10) c.f.u./ml. At the high nutrient level, biofilm EPS content and DHA both showed the increasing trend with the increase of suspended bacterial count. This work provides the basis and reference for management strategies in actual recirculating cooling water systems. PMID:22805808
Liu, Fang; Dong, Wenwen; Yang, Fei; Lu, Lei; Wang, Yongqiang; Yin, Xiaochen; Zhao, Chaocheng
Tannins Possessing Bacteriostatic Effect Impair Pseudomonas aeruginosa Adhesion and Biofilm Formation
Plants produce many compounds that are biologically active, either as part of their normal program of growth and development or in response to pathogen attack or stress. Traditionally, Anadenanthera colubrina, Commiphora leptophloeos and Myracrodruon urundeuva have been used by communities in the Brazilian Caatinga to treat several infectious diseases. The ability to impair bacterial adhesion represents an ideal strategy to combat bacterial pathogenesis, because of its importance in the early stages of the infectious process; thus, the search for anti-adherent compounds in plants is a very promising alternative. This study investigated the ability of stem-bark extracts from these three species to control the growth and prevent biofilm formation of Pseudomonas aeruginosa, an important opportunistic pathogen that adheres to surfaces and forms protective biofilms. A kinetic study (0�72 h) demonstrated that the growth of extract-treated bacteria was inhibited up to 9 h after incubation, suggesting a bacteriostatic activity. Transmission electron microscopy and fluorescence microscopy showed both viable and nonviable cells, indicating bacterial membrane damage; crystal violet assay and scanning electron microscopy demonstrated that treatment strongly inhibited biofilm formation during 6 and 24 h and that matrix production remained impaired even after growth was restored, at 24 and 48 h of incubation. Herein, we propose that the identified (condensed and hydrolyzable) tannins are able to inhibit biofilm formation via bacteriostatic properties, damaging the bacterial membrane and hindering matrix production. Our findings demonstrate the importance of this abundant class of Natural Products in higher plants against one of the most challenging issues in the hospital setting: biofilm resilience.
Trentin, Danielle S.; Silva, Denise B.; Amaral, Matheus W.; Zimmer, Karine R.; Silva, Marcia V.; Lopes, Norberto P.; Giordani, Raquel B.; Macedo, Alexandre J.
Tannins possessing bacteriostatic effect impair Pseudomonas aeruginosa adhesion and biofilm formation.
Plants produce many compounds that are biologically active, either as part of their normal program of growth and development or in response to pathogen attack or stress. Traditionally, Anadenanthera colubrina, Commiphora leptophloeos and Myracrodruon urundeuva have been used by communities in the Brazilian Caatinga to treat several infectious diseases. The ability to impair bacterial adhesion represents an ideal strategy to combat bacterial pathogenesis, because of its importance in the early stages of the infectious process; thus, the search for anti-adherent compounds in plants is a very promising alternative. This study investigated the ability of stem-bark extracts from these three species to control the growth and prevent biofilm formation of Pseudomonas aeruginosa, an important opportunistic pathogen that adheres to surfaces and forms protective biofilms. A kinetic study (0-72 h) demonstrated that the growth of extract-treated bacteria was inhibited up to 9 h after incubation, suggesting a bacteriostatic activity. Transmission electron microscopy and fluorescence microscopy showed both viable and nonviable cells, indicating bacterial membrane damage; crystal violet assay and scanning electron microscopy demonstrated that treatment strongly inhibited biofilm formation during 6 and 24 h and that matrix production remained impaired even after growth was restored, at 24 and 48 h of incubation. Herein, we propose that the identified (condensed and hydrolyzable) tannins are able to inhibit biofilm formation via bacteriostatic properties, damaging the bacterial membrane and hindering matrix production. Our findings demonstrate the importance of this abundant class of Natural Products in higher plants against one of the most challenging issues in the hospital setting: biofilm resilience. PMID:23776646
Trentin, Danielle S; Silva, Denise B; Amaral, Matheus W; Zimmer, Karine R; Silva, M�rcia V; Lopes, Norberto P; Giordani, Raquel B; Macedo, Alexandre J
Characterising the flux of carbon between calcium carbonate substrata, aqueous fluids, bacteria and a biofilm matrix
NASA Astrophysics Data System (ADS)
Quantification of the flux of elements between minerals, biofilms and aqueous solutions is essential in order to fully elucidate the role of microorganisms in mass transfer processes. Bacteria play a fundamental role in nearly all life and environmental processes and are by far the most abundant organisms on the planet. Their main mode of growth is in the form of biofilms growing on surfaces and although extensive study has been carried out into the problems biofilms cause to humans, the effects of biofilms in the environment are still poorly understood. Much of the published research describing biofilm growth on geological substrata emphasises the species present rather than their effect on the substratum. Although previous studies have shown that the production of organic or mineral acids by microbial biofilms can cause dissolution of mineral substrata, there is little quantitative data on the specific flux of elements between minerals, microbial cells, biofilm matrix and aqueous solutions. With growing evidence that microbial life occurs in abundance in the subsurface, biofilm activity within buried rocks and sediments may have important implications for global geochemical cycling of specific elements. Results will be presented as part of an ongoing laboratory analog experiment which has been designed to quantify the flux of carbon between all matrices in a laboratory substratum-biofilm-aqueous system. Water collected from a limestone cave system (pH 7.5-8; viable count 4.7�10^3 - 2.0�10^4 cfu.ml-1{) is used as innoculum and nutrient source for growing mixed consortium biofilms on a natural calcite (CaCO_3) substratum in a flow-through reaction cell (24 days; 25^oC; 1ml.min^{-1). Portions of the biofilm are periodically removed for microbiological analysis and the mineral surface microscopically examined for extent of alteration. A parallel experiment utilises an isotopically labelled synthetic 13C-CaCO_3 substratum. This enables carbon to be traced from the substratum, through the biofilm and into the aqueous and gas phase by analysing 13C/12C isotopic ratios in all system components. Through this mass-balance approach it is possible to quantify, for the first time, the carbon fluxes into the biofilm and the aqueous solution from the carbonate mineral substrate.
Rankin, S. C.; Cooke, D. A.; Handley, P. S.; Merrifield, C. M.; Wogelius, R. A.
Inhibition of biofilm formation by esomeprazole in Pseudomonas aeruginosa and Staphylococcus aureus.
Staphylococcus aureus and Pseudomonas aeruginosa are common nosocomial pathogens responsible for biofilm-associated infections. Proton pump inhibitors (PPI), such as esomeprazole, may have novel antimicrobial properties. The objective of this study was to assess whether esomeprazole prevents sessile bacterial growth and biofilm formation and whether it may have synergistic killing effects with standard antibiotics. The antibiofilm activity of esomeprazole at 0.25 mM was tested against two strains each of S. aureus and P. aeruginosa. Bacterial biofilms were prepared using a commercially available 96-peg-plate Calgary biofilm device. Sessile bacterial CFU counts and biomass were assessed during 72 hours of esomeprazole exposure. The killing activities after an additional 24 hours of vancomycin (against S. aureus) and meropenem (against P. aeruginosa) treatment with or without preexposure to esomeprazole were also assessed by CFU and biomass analyses. P. aeruginosa and S. aureus strains exposed to esomeprazole displayed decreased sessile bacterial growth and biomass (P < 0.001, each parameter). After 72 h of exposure, there was a 1-log(10) decrease in the CFU/ml of esomeprazole-exposed P. aeruginosa and S. aureus strains compared to controls (P < 0.001). After 72 h of exposure, measured absorbance was 100% greater in P. aeruginosa control strains than in esomeprazole-exposed strains (P < 0.001). Increased killing and decreased biomass were observed for esomeprazole-treated bacteria compared to untreated controls exposed to conventional antibiotics (P < 0.001, each parameter). Reduced biofilm growth after 24 h was visibly apparent by light micrographs for P. aeruginosa and S. aureus isolates exposed to esomeprazole compared to untreated controls. In conclusion, esomeprazole demonstrated an antibiofilm effect against biofilm-producing S. aureus and P. aeruginosa. PMID:22664967
Singh, Vandana; Arora, Vaneet; Alam, M Jahangir; Garey, Kevin W
Inhibition of Biofilm Formation by Esomeprazole in Pseudomonas aeruginosa and Staphylococcus aureus
Staphylococcus aureus and Pseudomonas aeruginosa are common nosocomial pathogens responsible for biofilm-associated infections. Proton pump inhibitors (PPI), such as esomeprazole, may have novel antimicrobial properties. The objective of this study was to assess whether esomeprazole prevents sessile bacterial growth and biofilm formation and whether it may have synergistic killing effects with standard antibiotics. The antibiofilm activity of esomeprazole at 0.25 mM was tested against two strains each of S. aureus and P. aeruginosa. Bacterial biofilms were prepared using a commercially available 96-peg-plate Calgary biofilm device. Sessile bacterial CFU counts and biomass were assessed during 72 hours of esomeprazole exposure. The killing activities after an additional 24 hours of vancomycin (against S. aureus) and meropenem (against P. aeruginosa) treatment with or without preexposure to esomeprazole were also assessed by CFU and biomass analyses. P. aeruginosa and S. aureus strains exposed to esomeprazole displayed decreased sessile bacterial growth and biomass (P < 0.001, each parameter). After 72 h of exposure, there was a 1-log10 decrease in the CFU/ml of esomeprazole-exposed P. aeruginosa and S. aureus strains compared to controls (P < 0.001). After 72 h of exposure, measured absorbance was 100% greater in P. aeruginosa control strains than in esomeprazole-exposed strains (P < 0.001). Increased killing and decreased biomass were observed for esomeprazole-treated bacteria compared to untreated controls exposed to conventional antibiotics (P < 0.001, each parameter). Reduced biofilm growth after 24 h was visibly apparent by light micrographs for P. aeruginosa and S. aureus isolates exposed to esomeprazole compared to untreated controls. In conclusion, esomeprazole demonstrated an antibiofilm effect against biofilm-producing S. aureus and P. aeruginosa.
Singh, Vandana; Arora, Vaneet; Alam, M. Jahangir
Triclosan causes toxic effects to algae in marine biofilms, but does not inhibit the metabolic activity of marine biofilm bacteria.
Effects of the antimicrobial agent triclosan to natural periphyton communities (biofilms, comprising primarily microalgae and bacteria) were assessed in two independent experiments during spring and summer. For that purpose a semi-static test system was used in which periphyton was exposed to a concentration range of 5-9054nmol/L triclosan. Effects on algae were analyzed as content and composition of photosynthetic pigments. The corresponding EC50 values were 39.25 and 302.45nmol/L for the spring and summer experiment, respectively. Effects on periphytic bacteria were assessed as effects on carbon utilization patterns, using Biolog Ecoplates. No inhibition of either total carbon utilization or functional diversity was observed, indicating a pronounced triclosan tolerance of the marine bacteria. In contrast, a small stimulation of the total carbon utilization was observed at triclosan concentrations exceeding 100nmol/L. PMID:24928457
Johansson, C Henrik; Janmar, Lisa; Backhaus, Thomas
Role of Extracellular DNA during Biofilm Formation by Listeria monocytogenes? �
Listeria monocytogenes is a food-borne pathogen that is capable of living in harsh environments. It is believed to do this by forming biofilms, which are surface-associated multicellular structures encased in a self-produced matrix. In this paper we show that in L. monocytogenes extracellular DNA (eDNA) may be the only central component of the biofilm matrix and that it is necessary for both initial attachment and early biofilm formation for 41 L. monocytogenes strains that were tested. DNase I treatment resulted in dispersal of biofilms, not only in microtiter tray assays but also in flow cell biofilm assays. However, it was also demonstrated that in a culture without eDNA, neither Listeria genomic DNA nor salmon sperm DNA by itself could restore the capacity to adhere. A search for additional necessary components revealed that peptidoglycan (PG), specifically N-acetylglucosamine (NAG), interacted with the DNA in a manner which restored adhesion. If a short DNA fragment (less than approximately 500 bp long) was added to an eDNA-free culture prior to addition of genomic or salmon sperm DNA, adhesion was prevented, indicating that high-molecular-weight DNA is required for adhesion and that the number of attachment sites on the cell surface can be saturated.
Harmsen, Morten; Lappann, Martin; Kn?chel, Susanne; Molin, S?ren
Potentials of Mouthwashes in Disinfecting Cariogenic Bacteria and Biofilms Leading to Inhibition of Caries
Objectives: The aim of this study was to compare the effects of certain commercially available mouthwashes on cariogenic bacteria and biofilms, following the acquisition of inhibition potentials of caries. Materials and Methods: Mouthwashes containing I) chlorhexidine gluconate (CHG; 0.0005% w/v), II) benzethonium chloride (BTC; 0.01% w/v), III) an essential oil (Listerine), and IV) povidone-iodine (PVP-I; 0.035% w/v) were tested on planktonic cariogenic bacteria, biofilms, and an ex vivo caries model. Bacterial aliquots were inoculated with each solution separately and vortexed for 10 seconds at room temperature. Bacterial viability was subsequently investigated by fluorescence microscopy (FM) after staining with a BacLight viability kit and the number of colony-forming units (CFUs) was counted. Similarly, mouthwash solutions were applied to artificial cariogenic biofilms, and bacterial viability of the biofilms was investigated as stated above. Inhibition potentials of two selected mouthwashes of carious lesions were investigated using biofilm-induced caries and a secondary caries model. In all steps, a phosphate-buffered saline (PBS) solution was included as a control. Results: Planktonic cariogenic bacteria and bacteria embedded in biofilms were killed in remarkably large numbers with Listerine and PVP-I treatment compared to PBS and other gargles. CFU counts also showed significant reduction after treatment with Listerine and PVP-I compared to other solutions (P<0.05). Listerine also displayed significant (P<0.05) inhibition effects in preventing the progression of demineralization. Conclusion: Bactericidal potencies of the mouthwashes varied significantly, suggesting that mouthwashes like Listerine can be useful for the prevention of caries and secondary caries.
Oyanagi, Takehiro; Tagami, Junji; Matin, Khairul
Combination of Tigecycline and N-Acetylcysteine Reduces Biofilm-Embedded Bacteria on Vascular Catheters?
To assess the efficacy of an antibiofilm/antimicrobial agent combination, we incubated catheter segments colonized with one of six studied bacterial organisms in N-acetylcysteine, tigecycline, N-acetylcysteine-tigecycline, or saline. Segments were washed, sonicated, and cultured. N-acetylcysteine-tigecycline significantly decreased all viable biofilm-associated bacteria and was synergistic for methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis.
Aslam, Saima; Trautner, Barbara W.; Ramanathan, Venkat; Darouiche, Rabih O.
Thermophilic and mesophilic bacteria in biofilms associated with corrosion in a heat exchanger
Biofilm development on AISI-1020 carbon steel coupons installed at the outlet of a heat exchanger was evaluated at the thirtieth and the sixtieth days of exposure. Water temperature varied between 41 and 60?�C. The most probable number technique (MPN) was applied to quantify mesophilic and thermophilic species of aerobic, anaerobic, and sulphate-reducing bacteria (SRB) in planktonic and sessile phases. The
M. A. N. Almeida; F. P. de Fran�a
Capillary isoelectric focusing--useful tool for detection of the biofilm formation in Staphylococcus epidermidis.
The biofilm formation is an important factor of S. epidermidis virulence. Biofilm-positive strains might be clinically more important than biofilm-negative ones. Unlike biofilm-negative staphylococci, biofilm-positive staphylococci are surrounded with an extracellular polysaccharide substance. The presence of this substance on the surface can affect physico-chemical properties of the bacterial cell, including surface charge. 73 S. epidermidis strains were examined for the presence of ica operon, for the ability to form biofilm by Christensen test tube method and for the production of slime by Congo red agar method. Isoelectric points (pI) of these strains were determined by means of Capillary Isoelectric Focusing. The biofilm negative strains focused near pI value 2.3, while the pI values of the biofilm positive strains were near 2.6. Isoelectric point is a useful criterion for the differentiation between biofilm-positive and biofilm-negative S. epidermidis strains. PMID:17157942
Ruzicka, Filip; Horka, Marie; Hola, Veronika; Votava, Miroslav
Synthetic quorum-sensing circuit to control consortial biofilm formation and dispersal in a microfluidic device
To utilize biofilms for chemical transformations in biorefineries they need to be controlled and replaced. Previously, we engineered the global regulator Hha and cyclic diguanylate-binding BdcA to create proteins that enable biofilm dispersal. Here we report a biofilm circuit that utilizes these two dispersal proteins along with a population-driven quorum-sensing switch. With this synthetic circuit, in a novel microfluidic device, we form an initial colonizer biofilm, introduce a second cell type (dispersers) into this existing biofilm, form a robust dual-species biofilm and displace the initial colonizer cells in the biofilm with an extracellular signal from the disperser cells. We also remove the disperser biofilm with a chemically induced switch, and the consortial population could tune. Therefore, for the first time, cells have been engineered that are able to displace an existing biofilm and then be removed on command allowing one to control consortial biofilm formation for various applications.
Hong, Seok Hoon; Hegde, Manjunath; Kim, Jeongyun; Wang, Xiaoxue; Jayaraman, Arul; Wood, Thomas K.
Synthetic quorum-sensing circuit to control consortial biofilm formation and dispersal in a microfluidic device.
To utilize biofilms for chemical transformations in biorefineries they need to be controlled and replaced. Previously, we engineered the global regulator Hha and cyclic diguanylate-binding BdcA to create proteins that enable biofilm dispersal. Here we report a biofilm circuit that utilizes these two dispersal proteins along with a population-driven quorum-sensing switch. With this synthetic circuit, in a novel microfluidic device, we form an initial colonizer biofilm, introduce a second cell type (dispersers) into this existing biofilm, form a robust dual-species biofilm and displace the initial colonizer cells in the biofilm with an extracellular signal from the disperser cells. We also remove the disperser biofilm with a chemically induced switch, and the consortial population could tune. Therefore, for the first time, cells have been engineered that are able to displace an existing biofilm and then be removed on command allowing one to control consortial biofilm formation for various applications. PMID:22215088
Hong, Seok Hoon; Hegde, Manjunath; Kim, Jeongyun; Wang, Xiaoxue; Jayaraman, Arul; Wood, Thomas K
Air-liquid interface biofilms of Bacillus cereus: formation, sporulation, and dispersion
Biofilm formation by Bacillus cereus was assessed using 56 strains of B. cereus, including the two sequenced strains, ATCC 14579 and ATCC 10987. Biofilm production in microtiter plates was found to be strongly dependent on incubation time, temperature, and medium, as well as the strain used, with some strains showing biofilm formation within 24 h and subsequent dispersion within the
Janneke G. E. Wijman; Leeuw de P. P. L. A; Roy Moezelaar; Marcel H. Zwietering; Tjakko Abee
Spontaneous switch to PIA-independent biofilm formation in an ica-positive Staphylococcus epidermidis isolate
The ability to form biofilms on abiotic surfaces is considered a major step in Staphylococcus epidermidis pathogenesis. In the majority of isolates, biofilm formation is mediated by the production of the polysaccharide intercellular adhesin PIA which is synthesized by enzymes encoded by the ica operon. Here, we report on a spontaneous switch to proteinaceous biofilm formation in an S. epidermidis
Susanne Hennig; Sun Nyunt Wai; Wilma Ziebuhr
Hypoxic Adaptation by Efg1 Regulates Biofilm Formation by Candida albicans
Hypoxia is encountered frequently by Candida albicans during systemic infection of the human host. We tested if hypoxia allows biofilm formation by C. albicans, which is a major cause of perseverance and antifungal resistance in C. albicans infections. Using an in vitro biofilm system, we unexpectedly discovered that several positive regulators of biofilm formation during normoxia, including Tec1, Ace2, Czf1,
Catrin Stichternoth; Joachim F. Ernst
Short communication Biofilm formation and the survival of Salmonella Typhimurium on parsley
Although several studies provide evidence that the formation of biofilms by human pathogens on plant tissue is possible, to date there is no direct evidence that biofilms enhance the resistance of plant-associated pathogens to disinfectants or biocides. We hypothesized that biofilm formation would enhance the adhesion and survival of Salmonella on leafy vegetables. To test our hypothesis, we compared the
Anat Lapidot; Ute Romling; Sima Yaron
Patterns of biofilm formation in intermittent and permanent streams: analysis of biofilm structure and metabolism
NASA Astrophysics Data System (ADS)
The development and functioning of benthic microbial communities in streams is largely dependent on the hydrological conditions. Climate change projections predict that the hydrological characteristics will probably be affected because of the rainfall regime. Hence, rivers from the Mediterranean region will become more similar to those draining arid or desert regions, while temperate streams will suffer of higher water flow fluctuations. In this study, we compared the process of biofilm formation between an intermittent (the Fuirosos, Mediterranean) and a permanent (the Walzbach, Central European) stream. Specifically, we analyzed the succession of bacterial and algal populations in the biofilm through bacterial rDNA sequences analysis (16S rDNA and 16S-23S intergenic sequence) and diatom taxa identification over a 60-days colonization experiment. Moreover, changes in biofilm structural (microbial biomass and extracellular polysaccharide content) and metabolic (extracellular enzyme activities) parameters were also analyzed. The successional patterns of microbial populations in the Fuirosos showed clear discontinutities coinciding with flood episodes while at the Walzbach the time sequence was more gradual. Although both study sites were forested, greater microbial biomass standing stock (algal and bacterial) and greater species biodiversity was detected during biofilm development at the Mediterranean site. The higher bacterial biodiversity may be related to the potential effect of flooding episodes in reducing biological interactions in complex microbial communities, such as the competitive exclusion of species. Moreover, the presence of rapid colonizing diatom species might be an adaptation to hydrological changes. In contrast, species competition could define the more stable environments, such as that observed in the Central European stream. Overall, the hystorical evolutionary pressure from the different bioclimatic regions could be also affecting the microbial community composition. Further, the analysis of the whole biofilm colonization sequence showed a greater EPS development and a higher potential extracellular enzyme activity rates in the Mediterranean stream. The higher EPS reflected a functional response of the biofilms to avoid detachment during high flow episodes and the higher enzyme activities were according to the low water nutrient concentration available for microbes. However, the higher nutrient conditions (higher nitrate and phosphate concentrations) at the Central European stream was not reflected on higher biomass standing stock in biofilms. Likely, the biomass accrual of biofilms was related to the differences in climatic conditions between study sites (e. g. daily insulation, temperature oscillation). The results suggest that microbial community development and functioning is primarily related to the physical characteristics of the different sites (specially the stream hydrology, but also the light regime).
Artigas, J.; Schwartz, T.; Kirchen, S.; Roman�, A. M.; Fund, K.; Obst, U.; Sabater, S.
Structural Basis for Biofilm Formation via the Vibrio cholerae Matrix Protein RbmA
During the transition from a free-swimming, single-cell lifestyle to a sessile, multicellular state called a biofilm, bacteria produce and secrete an extracellular matrix comprised of nucleic acids, exopolysaccharides, and adhesion proteins. The Vibrio cholerae biofilm matrix contains three major protein components, RbmA, Bap1, and RbmC, which are unique to Vibrio cholerae and appear to support biofilm formation at particular steps in the process. Here, we focus on RbmA, a structural protein with an unknown fold. RbmA participates in the early cell-cell adhesion events and is found throughout the biofilm where it localizes to cell-cell contact sites. We determined crystal structures of RbmA and revealed that the protein folds into tandem fibronectin type III (FnIII) folds. The protein is dimeric in solution and in crystals, with the dimer interface displaying a surface groove that is lined with several positively charged residues. Structure-guided mutagenesis studies establish a crucial role for this surface patch for RbmA function. On the basis of the structure, we hypothesize that RbmA serves as a tether by maintaining flexible linkages between cells and the extracellular matrix.
Giglio, Krista M.; Fong, Jiunn C.
High Levels of Genetic Recombination during Nasopharyngeal Carriage and Biofilm Formation in Streptococcus pneumoniae
ABSTRACT Transformation of genetic material between bacteria was first observed in the 1920s using Streptococcus pneumoniae as a model organism. Since then, the mechanism of competence induction and transformation has been well characterized, mainly using planktonic bacteria or septic infection models. However, epidemiological evidence suggests that genetic exchange occurs primarily during pneumococcal nasopharyngeal carriage, which we have recently shown is associated with biofilm growth, and is associated with cocolonization with multiple strains. However, no studies to date have comprehensively investigated genetic exchange during cocolonization in vitro and in vivo or the role of the nasopharyngeal environment in these processes. In this study, we show that genetic exchange during dual-strain carriage in vivo is extremely efficient (10?2) and approximately 10,000,000-fold higher than that measured during septic infection (10?9). This high transformation efficiency was associated with environmental conditions exclusive to the nasopharynx, including the lower temperature of the nasopharynx (32 to 34�C), limited nutrient availability, and interactions with epithelial cells, which were modeled in a novel biofilm model in vitro that showed similarly high transformation efficiencies. The nasopharyngeal environmental factors, combined, were critical for biofilm formation and induced constitutive upregulation of competence genes and downregulation of capsule that promoted transformation. In addition, we show that dual-strain carriage in vivo and biofilms formed in vitro can be transformed during colonization to increase their pneumococcal fitness and also, importantly, that bacteria with lower colonization ability can be protected by strains with higher colonization efficiency, a process unrelated to genetic exchange.
Marks, Laura R.; Reddinger, Ryan M.; Hakansson, Anders P.
Early microbial biofilm formation on marine plastic debris.
An important aspect of the global problem of plastic debris pollution is plastic buoyancy. There is some evidence that buoyancy is influenced by attached biofilms but as yet this is poorly understood. We submerged polyethylene plastic in seawater and sampled weekly for 3 weeks in order to study early stage processes. Microbial biofilms developed rapidly on the plastic and coincided with significant changes in the physicochemical properties of the plastic. Submerged plastic became less hydrophobic and more neutrally buoyant during the experiment. Bacteria readily colonised the plastic but there was no indication that plastic-degrading microorganisms were present. This study contributes to improved understanding of the fate of plastic debris in the marine environment. PMID:21093883
Lobelle, Delphine; Cunliffe, Michael
High-Throughput Screening of Multispecies Biofilm Formation and Quantitative PCR-Based Assessment of Individual Species Proportions, Useful for Exploring Interspecific Bacterial Interactions.
Multispecies biofilms are predominant in almost all natural environments, where myriads of resident microorganisms interact with each other in both synergistic and antagonistic manners. The interspecies interactions among different bacteria are, despite the ubiquity of these communities, still poorly understood. Here, we report a rapid, reproducible and sensitive approach for quantitative screening of biofilm formation by bacteria when cultivated as mono- and multispecies biofilms, based on the Nunc-TSP lid system and crystal violet staining. The relative proportion of the individual species in a four-species biofilm was assessed using quantitative PCR based on SYBR Green I fluorescence with specific primers. The results indicated strong synergistic interactions in a four-species biofilm model community with a more than 3-fold increase in biofilm formation and demonstrated the strong dominance of two strains, Xanthomonas retroflexus and Paenibacillus amylolyticus. The developed approach can be used as a standard procedure for evaluating interspecies interactions in defined microbial communities. This will be of significant value in the quantitative study of the microbial composition of multispecies biofilms both in natural environments and infectious diseases to increase our understanding of the mechanisms that underlie cooperation, competition and fitness of individual species in mixed-species biofilms. PMID:24337804
Ren, Dawei; Madsen, Jonas Stenl�kke; de la Cruz-Perera, Claudia I; Bergmark, Lasse; S�rensen, S�ren J; Burm�lle, Mette
The Role of Antimicrobial Peptides in Preventing Multidrug-Resistant Bacterial Infections and Biofilm Formation
Over the last decade, decreasing effectiveness of conventional antimicrobial-drugs has caused serious problems due to the rapid emergence of multidrug-resistant pathogens. Furthermore, biofilms, which are microbial communities that cause serious chronic infections and dental plaque, form environments that enhance antimicrobial resistance. As a result, there is a continuous search to overcome or control such problems, which has resulted in antimicrobial peptides being considered as an alternative to conventional drugs. Antimicrobial peptides are ancient host defense effector molecules in living organisms. These peptides have been identified in diverse organisms and synthetically developed by using peptidomimic techniques. This review was conducted to demonstrate the mode of action by which antimicrobial peptides combat multidrug-resistant bacteria and prevent biofilm formation and to introduce clinical uses of these compounds for chronic disease, medical devices, and oral health. In addition, combinations of antimicrobial peptides and conventional drugs were considered due to their synergetic effects and low cost for therapeutic treatment.
Park, Seong-Cheol; Park, Yoonkyung; Hahm, Kyung-Soo
A different path: revealing the function of staphylococcal proteins in biofilm formation.
Staphylococcus aureus and Staphylococcus epidermidis cause dangerous and difficult to treat medical device-related infections through their ability to form biofilms. Extracellular poly-N-acetylglucosamine (PNAG) facilitates biofilm formation and is a vaccination target, yet details of its biosynthesis by the icaADBC gene products is limited. IcaC is the proposed transporter for PNAG export, however a comparison of the Ica proteins to homologous exo-polysaccharide synthases suggests that the common IcaAD protein components both synthesise and transport the PNAG. The limited distribution of icaC to the Staphylococcaceae and its membership of a family of membrane-bound acyltransferases, leads us to suggest that IcaC is responsible for the known O-succinylation of PNAG that occurs in staphylococci, identifying a potentially new therapeutic target specific for these bacteria. PMID:24735724
Atkin, Kate E; MacDonald, Sandy J; Brentnall, Andrew S; Potts, Jennifer R; Thomas, Gavin H
Role of ClpP in biofilm formation and virulence of Staphylococcus epidermidis
Infections caused by the leading nosocomial pathogen Staphylococcus epidermidis are characterized by biofilm formation on implanted medical devices. However, the molecular basis of biofilm formation and its regulation are not completely understood. Here, we describe an important role of the ClpP protease in biofilm development and virulence of S. epidermidis. We constructed an isogenic clpP mutant strain of a biofilm-forming
Chongzhen Wang; Min Li; Dandan Dong; Jianping Wang; Jun Ren; Michael Otto; Qian Gao
Cholate-Stimulated Biofilm Formation by Lactococcus lactis Cells ? �
Bile acid resistance by Lactococcus lactis depends on the ABC-type multidrug transporter LmrCD. Upon deletion of the lmrCD genes, cells can reacquire bile acid resistance upon prolonged exposure to cholate, yielding the ?lmrCDr strain. The resistance mechanism in this strain is non-transporter based. Instead, cells show a high tendency to flocculate, suggesting cell surface alterations. Contact angle measurements demonstrate that the ?lmrCDr cells are equipped with an increased cell surface hydrophilicity compared to those of the parental and wild-type strains, while the surface hydrophilicity is reduced in the presence of cholate. ?lmrCDr cells are poor in biofilm formation on a hydrophobic polystyrene surface, but in the presence of subinhibitory concentrations of cholate, biofilm formation is strongly stimulated. Biofilm cells show an enhanced extracellular polymeric substance production and are highly resistant to bile acids. These data suggest that non-transporter-based cholate resistance in L. lactis is due to alterations in the cell surface that stimulate cells to form resistant biofilms.
Zaidi, Arsalan Haseeb; Bakkes, Patrick J.; Krom, Bastiaan P.; van der Mei, Henny C.; Driessen, Arnold J. M.
Staphylococcal biofilm formation as affected by type acidulant.
Staphylococcal growth and biofilm formation in culture medium where pH was lowered with weak organic (acetic and lactic) or strong inorganic (hydrochloric) acids were studied. The effects were evaluated by biomass measurements, cell-surface hydrophobicity, scanning electron microscopy (SEM), and confocal laser scanning microscopy (CLSM). The results demonstrated that the inhibition was related to type of acidulant and pH value. At pH 5.0, the antibacterial effect was more pronounced in the presence of acetic acid (58-60% growth reduction) compared with that in the presence of lactic (7-16% growth reduction) and hydrochloric acids (23-24% reduction). The biofilm biomass of Staphylococcus aureus and Staphylococcus epidermidis was reduced by 92, 85, 63, and 93, 87, 81% after exposition to acetic, lactic, and hydrochloric acids, respectively. Increasing the pH from 5.0 to 6.0 resulted in a noticeable reduction in the effectiveness of acids. A minor cells hydrophobic character was also documented. The SEM and CLSM revealed a poorly structured and thinner biofilm compared with the dense and multilayered control. Acidic environment could have important implications for food-processing system to prevent bacterial colonization and control biofilm formation. The findings of this study lead to consider the rational use of the type of acid to achieve acidic environments. PMID:24320800
Nostro, Antonia; Cellini, Luigina; Ginestra, Giovanna; D'Arrigo, Manuela; di Giulio, Mara; Marino, Andreana; Blanco, Anna Rita; Favaloro, Angelo; Bisignano, Giuseppe
Characterization of biofilm formation by Borrelia burgdorferi in vitro.
Borrelia burgdorferi, the causative agent of Lyme disease, has long been known to be capable of forming aggregates and colonies. It was recently demonstrated that Borrelia burgdorferi aggregate formation dramatically changes the in vitro response to hostile environments by this pathogen. In this study, we investigated the hypothesis that these aggregates are indeed biofilms, structures whose resistance to unfavorable conditions are well documented. We studied Borrelia burgdorferi for several known hallmark features of biofilm, including structural rearrangements in the aggregates, variations in development on various substrate matrices and secretion of a protective extracellular polymeric substance (EPS) matrix using several modes of microscopic, cell and molecular biology techniques. The atomic force microscopic results provided evidence that multilevel rearrangements take place at different stages of aggregate development, producing a complex, continuously rearranging structure. Our results also demonstrated that Borrelia burgdorferi is capable of developing aggregates on different abiotic and biotic substrates, and is also capable of forming floating aggregates. Analyzing the extracellular substance of the aggregates for potential exopolysaccharides revealed the existence of both sulfated and non-sulfated/carboxylated substrates, predominately composed of an alginate with calcium and extracellular DNA present. In summary, we have found substantial evidence that Borrelia burgdorferi is capable of forming biofilm in vitro. Biofilm formation by Borrelia species might play an important role in their survival in diverse environmental conditions by providing refuge to individual cells. PMID:23110225
Sapi, Eva; Bastian, Scott L; Mpoy, Cedric M; Scott, Shernea; Rattelle, Amy; Pabbati, Namrata; Poruri, Akhila; Burugu, Divya; Theophilus, Priyanka A S; Pham, Truc V; Datar, Akshita; Dhaliwal, Navroop K; MacDonald, Alan; Rossi, Michael J; Sinha, Saion K; Luecke, David F
Capsaicin inhibits Porphyromonas gingivalis growth, biofilm formation, gingivomucosal inflammatory cytokine secretion, and in vitro osteoclastogenesis.
The prevention and treatment of periodontitis requires not only the control of causative pathogens, especially Porphyromonas gingivalis, but also the regulation of inflammatory immune response. Investigating auxiliary drugs for periodontitis during conventional treatments is, thus, quite important. Capsaicin, an agonist for the vanilloid receptor subtype 1 (TRPV1), due to its bacteriostatic activity against Gram-negative bacteria and anti-inflammatory effects, appears to be a promising drug. In this work, the antimicrobial activity of capsaicin against P. gingivalis and biofilm formation, inflammatory cytokine levels in experimental periodontitis, osteoclast precursor proliferation, and osteoclastogenesis in vitro were fully investigated. The results showed that capsaicin inhibited P. gingivalis growth with a minimum inhibitory concentration (MIC) and a minimum bactericidal concentration (MBC) of 16 and 64 mg/l, respectively. Capsaicin also inhibited P. gingivalis biofilm formation, with minimum biofilm inhibition concentrations MBIC50 and MBIC90 of 16 and 32 mg/l, respectively, and reduced pre-formed biofilms' viability with a minimum biofilm reduction concentration MBRC50 of 64 mg/l, as demonstrated by confocal laser scanning microscopy. In experimental periodontitis, except for IL-10, TNF-?, IL-1?, IL-6, IL-12, and iNOS were depressed after capsaicin treatment. Moreover, capsaicin also suppressed osteoclast precursor proliferation and osteoclastogenesis, as demonstrated by NF-?B p65. However, this favorable effect was attenuated by the TRPV1 antagonist, camphor. It, thus, suggests that capsaicin is a potential drug for the auxiliary treatment of periodontitis. TRPV1 activation may involve in beneficial roles of capsaicin on periodontitis. PMID:23955115
Zhou, Y; Guan, X; Zhu, W; Liu, Z; Wang, X; Yu, H; Wang, H
Effect of nitrofurans and NO generators on biofilm formation by Pseudomonas aeruginosa PAO1 and Burkholderia cenocepacia 370.
Antibacterial drugs in the nitrofuran series, such as nitrofurazone, furazidin, nitrofurantoin and nifuroxazide, as well as the nitric oxide generators sodium nitroprusside and isosorbide mononitrate in concentrations that do not suppress bacterial growth, were shown to increase the capacity of pathogenic bacteria Pseudomonas aeruginosa PAO1 and Burkholderia cenocepacia 370 to form biofilms. At 25-100microg/ml, nitrofurans 2-2.5-fold enhanced biofilm formation of P. aeruginosa PAO1, and NO donors 3-6-fold. For B. cenocepacia 370, the enhancement was 2-5-fold (nitrofurans) and 4.5-fold (sodium nitroprusside), respectively. PMID:19460431
Zaitseva, Julia; Granik, Vladimir; Belik, Alexandr; Koksharova, Olga; Khmel, Inessa
A Role for Amyloid in Cell Aggregation and Biofilm Formation
Cell adhesion molecules in Saccharomyces cerevisiae and Candida albicans contain amyloid-forming sequences that are highly conserved. We have now used site-specific mutagenesis and specific peptide perturbants to explore amyloid-dependent activity in the Candida albicans adhesin Als5p. A V326N substitution in the amyloid-forming region conserved secondary structure and ligand binding, but abrogated formation of amyloid fibrils in soluble Als5p and reduced cell surface thioflavin T fluorescence. When displayed on the cell surface, Als5p with this substitution prevented formation of adhesion nanodomains and formation of large cellular aggregates and model biofilms. In addition, amyloid nanodomains were regulated by exogenous peptides. An amyloid-forming homologous peptide rescued aggregation and biofilm activity of Als5pV326N cells, and V326N substitution peptide inhibited aggregation and biofilm activity in Als5pWT cells. Therefore, specific site mutation, inhibition by anti-amyloid peturbants, and sequence-specificity of pro-amyloid and anti-amyloid peptides showed that amyloid formation is essential for nanodomain formation and activation.
Garcia, Melissa C.; Lee, Janis T.; Ramsook, Caleen B.; Alsteens, David; Dufrene, Yves F.; Lipke, Peter N.
Muramidases found in the foregut microbiome of the Tammar wallaby can direct cell aggregation and biofilm formation
We describe here the role of muramidases present in clones of metagenomic DNA that result in cell aggregation and biofilm formation by Escherichia coli. The metagenomic clones were obtained from uncultured Lachnospiraceae-affiliated bacteria resident in the foregut microbiome of the Tammar wallaby. One of these fosmid clones (p49C2) was chosen for more detailed studies and a variety of genetic methods were used to delimit the region responsible for the phenotype to an open reading frame of 1425?bp. Comparative sequence analysis with other fosmid clones giving rise to the same phenotype revealed the presence of muramidase homologues with the same modular composition. Phylogenetic analysis of the fosmid sequence data assigned these fosmid inserts to recently identified, but uncultured, phylogroups of Lachnospiraceae believed to be numerically dominant in the foregut microbiome of the Tammar wallaby. The muramidase is a modular protein containing putative N-acetylmuramoyl--alanine amidase and an endo-?-N-acetylglucosaminidase catalytic module, with a similar organization and functional properties to some Staphylococcal autolysins that also confer adhesive properties and biofilm formation. We also show here that the cloned muramidases result in the production of extracellular DNA, which appears to be the key for biofilm formation and autoaggregation. Collectively, these findings suggest that biofilm formation and cell aggregation in gut microbiomes might occur via the concerted action of carbohydrate-active enzymes and the production of extracellular DNA to serve as a biofilm scaffold.
Pope, Phillip B; Totsika, Makrina; Aguirre de Carcer, Daniel; Schembri, Mark A; Morrison, Mark
Fluid Flow Induces Biofilm Formation in Staphylococcus epidermidis Polysaccharide Intracellular Adhesin-Positive Clinical Isolates
Staphylococcus epidermidis is a common cause of catheter-related bloodstream infections, resulting in significant morbidity and mortality and increased hospital costs. The ability to form biofilms plays a crucial role in pathogenesis; however, not all clinical isolates form biofilms under normal in vitro conditions. Strains containing the ica operon can display significant phenotypic variation with respect to polysaccharide intracellular adhesin (PIA)-based biofilm formation, including the induction of biofilms upon environmental stress. Using a parallel microfluidic approach to investigate flow as an environmental signal for S. epidermidis biofilm formation, we demonstrate that fluid shear alone induces PIA-positive biofilms of certain clinical isolates and influences biofilm structure. These findings suggest an important role of the catheter microenvironment, particularly fluid flow, in the establishment of S. epidermidis infections by PIA-dependent biofilm formation.
Weaver, Westbrook M.; Miller, Jeff F.; Di Carlo, Dino
Efficacy of clarithromycin on biofilm formation of methicillin-resistant Staphylococcus pseudintermedius
Background Surgical site infections (SSIs) caused by biofilm-forming methicillin-resistant Staphylococcus pseudintermedius (MRSP) have emerged as the most common hospital-acquired infections in companion animals. No methods currently exist for the therapeutic remediation of SSIs caused by MRSP in biofilms. Clarithromycin (CLA) has been shown to prevent biofilm formation by Staphylococcus aureus. This study aims to assess the in vitro activity of CLA in eradicating MRSP biofilm formation on various materials. Results Quantitative assay results (P = 0.5126) suggest that CLA does not eradicate MRSP biofilm formation on polystyrene after 4 � 24 h growth periods. Scanning electron micrographs confirmed that CLA did not eradicate MRSP biofilm formed on orthopaedic implants. Conclusions By determining the in vitro characteristics and activities of MRSP isolates alone and against antibiotics, in vitro models of biofilm related infections can be made. In vitro data suggests that CLA does not effectively eradicate S. pseudintermedius biofilms in therapeutic doses.
Altered host cell-bacteria interaction due to nanoparticle interaction with a bacterial biofilm.
Nanoparticle (NP) use in everyday applications creates the potential for NPs to enter the environment where, in aquatic systems, they are likely to settle on substrates and interact with microbial communities. Legionella pneumophila biofilms are found as part of microbial communities in both natural and man-made environments, especially in man-made cooling systems. The bacterium is the causative agent of Legionnaires' disease. Legionella requires a host cell for replication in the environment, and amoebae commonly serve as this host cell. Our previous work demonstrated significant changes in Legionella biofilm morphology after exposure to 0.7 ?g/L gold NPs (AuNPs). Here, we investigate how these morphology changes alter host-bacteria interactions using Acanthamoeba polyphaga as a model. Host-bacteria-NP interactions are affected by NP characteristics. Biofilms exposed to 4- and 18-nm, citrate-capped, spherical AuNPs significantly altered the grazing ability of A. polyphaga, which was not observed in biofilms exposed to 24-nm polystyrene beads. Uptake and replication of NP-exposed planktonic L. pneumophila within A. polyphaga were not altered regardless of NP size or core chemistry. Nanomaterial effects on the interaction of benthic organisms and bacteria may be directly or, as shown here, indirectly dependent on bacterial morphology. NP contamination therefore may alter interactions in a normal ecosystem function. PMID:23052925
Raftery, Tara D; Lindler, Heidi; McNealy, Tamara L
Biofilm Formation and Sloughing in Serratia marcescens Are Controlled by Quorum Sensing and Nutrient Cues
We describe here a role for quorum sensing in the detachment, or sloughing, of Serratia marcescens filamentous biofilms, and we show that nutrient conditions affect the biofilm morphotype. Under reduced carbon or nitrogen conditions, S. marcescens formed a classical biofilm consisting of microcolonies. The filamentous biofilm could be converted to a microcolony-type biofilm by switching the medium after establishment of the biofilm. Similarly, when initially grown as a microcolony biofilm, S. marcescens could be converted back to a filamentous biofilm by increasing the nutrient composition. Under high-nutrient conditions, an N-acyl homoserine lactone quorum-sensing mutant formed biofilms that were indistinguishable from the wild-type biofilms. Similarly, other quorum-sensing-dependent behaviors, such as swarming motility, could be rendered quorum sensing independent by manipulating the growth medium. Quorum sensing was also found to be involved in the sloughing of the filamentous biofilm. The biofilm formed by the bacterium consistently sloughed from the substratum after approximately 75 to 80 h of development. The quorum-sensing mutant, when supplemented with exogenous signal, formed a wild-type filamentous biofilm and sloughed at the same time as the wild type, and this was independent of surfactant production. When we removed the signal from the quorum-sensing mutant prior to the time of sloughing, the biofilm did not undergo significant detachment. Together, the data suggest that biofilm formation by S. marcescens is a dynamic process that is controlled by both nutrient cues and the quorum-sensing system.
Rice, S. A.; Koh, K. S.; Queck, S. Y.; Labbate, M.; Lam, K. W.; Kjelleberg, S.
Human serum inhibits adhesion and biofilm formation in Candida albicans
Background Candida albicans can form biofilms on intravenous catheters; this process plays a key role in the pathogenesis of catheter infections. This study evaluated the effect of human serum (HS) on C. albicans biofilm formation and the expression of adhesion-related genes in vitro. A C. albicans laboratory strain (ATCC90028) and three clinical strains were grown for 24�h in RPMI 1640 supplemented with HS or RPMI 1640 alone (as a control). The growth of biofilm cells of four strains was monitored by a Live Cell Movie Analyzer, and by XTT reduction assay. The expression of the adhesion-related genes BCR1, ALS1, ALS3, HWP1 and ECE1 was analyzed by RT-PCR at three time points (60�min, 90�min, and 24�h). Results In the adhesion phase, C. albicans cells kept a Brownian movement in RPMI medium containing HS until a large number of germ tubes were formed. In the control group, C. albicans cells quickly adhered to the bottom of the reaction plate. Compared with RPMI 1640, medium supplemented with 3�50% HS caused a significant decrease in biofilm development (all p?0.001). However, the presence of HS had no significant inhibitory effect on the pre-adhered biofilms (all p?>?0.05). Biofilm formation was also inhibited by heat-inactivated and proteinase K pre-treated HS. The presence of 50% HS did not significantly affect the planktonic growth of C. albicans (p?>?0.05). At three time points, HS inhibited expression of the ALS1 and ALS3 genes and promoted expression of the HWP1 and ECE1 genes. Significant up-regulation of BCR1 was observed only at the 90-min point. Conclusions Human serum reduces biofilm formation by inhibiting the adhesion of C. albicans cells. This response may be associated with the down-regulation of adhesion-related genes ALS1, ALS3 and BCR1. The inhibitory serum component is protease-resistant and heat stable.
Biofilm comprising phototrophic, diazotrophic, and hydrocarbon-utilizing bacteria: a promising consortium in the bioremediation of aquatic hydrocarbon pollutants.
Biofilms harboring simultaneously anoxygenic and oxygenic phototrophic bacteria, diazotrophic bacteria, and hydrocarbon-utilizing bacteria were established on glass slides suspended in pristine and oily seawater. Via denaturing gradient gel electrophoresis analysis on PCR-amplified rRNA gene sequence fragments from the extracted DNA from biofilms, followed by band amplification, biofilm composition was determined. The biofilms contained anoxygenic phototrophs belonging to alphaproteobacteria; pico- and filamentous cyanobacteria (oxygenic phototrophs); two species of the diazotroph Azospirillum; and two hydrocarbon-utilizing gammaproteobacterial genera, Cycloclasticus and Oleibacter. The coexistence of all these microbial taxa with different physiologies in the biofilm makes the whole community nutritionally self-sufficient and adequately aerated, a condition quite suitable for the microbial biodegradation of aquatic pollutant hydrocarbons. PMID:23089957
Al-Bader, Dhia; Kansour, Mayada K; Rayan, Rehab; Radwan, Samir S
The influence of bacteria on struvite crystal habit and its importance in urinary stone formation
NASA Astrophysics Data System (ADS)
Infection-induced urinary stones form as a result of a urinary tract infection by urease-producing bacteria. These stones are not totally crystalline in nature but rather consist of an agglomeration of bacteria, organic matrix, and crystal of struvite (MgNH 4PO 4� 6H 2O). Crystal formation is related to the ability of the bacteria to effect an increase in the urine pH. Another equally important bacterial role lies in their formation of a 'biofilm' which later becomes the organic matrix constituent of the stone. Results of the present in vitro study indicate that crystals are formed more readily if produced within the bacterial biofilm than in the surrounding urine. It is proposed that supersaturation, due in part to a bacterial-induced pH increase and in part to the metal binding tendency of the biofilm, leads to crystal formation via a gel growth mechanism within the biofilm itself. In time further bacterial cell division, microcolony.
Clapham, L.; McLean, R. J. C.; Nickel, J. C.; Downey, J.; Costerton, J. W.
A coverslip-based technique for evaluating Staphylococcus aureus biofilm formation on human plasma
The ability of the opportunistic pathogen, Staphylococcus aureus, to form biofilms is increasingly being viewed as an important contributor to chronic infections. In vitro methods for analyzing S. aureus biofilm formation have focused on bacterial attachment and accumulation on abiotic surfaces, such as in microtiter plate and flow cell assays. Microtiter plates provide a rapid measure of relative biomass levels, while flow cells have limited experimental throughput but are superior for confocal microscopy biofilm visualization. Although these assays have proven effective at identifying mechanisms involved in cell attachment and biofilm accumulation, the significance of these assays in vivo remains unclear. Studies have shown that when medical devices are implanted they are coated with host factors, such as matrix proteins, that facilitate S. aureus attachment and biofilm formation. To address the challenge of integrating existing biofilm assay features with a biotic surface, we have established an in vitro biofilm technique utilizing UV-sterilized coverslips coated with human plasma. The substratum more closely resembles the in vivo state and provides a platform for S. aureus to establish a robust biofilm. Importantly, these coverslips are amenable to confocal microscopy imaging to provide a visual reference of the biofilm growth stage, effectively merging the benefits of the microtiter and flow cell assays. We confirmed the approach using clinical S. aureus isolates and mutants with known biofilm phenotypes. Altogether, this new biofilm assay can be used to assess the function of S. aureus virulence factors associated with biofilm formation and for monitoring the efficacy of biofilm treatment modalities.
Walker, Jennifer N.; Horswill, Alexander R.
Control of marine biofouling and medical biofilm formation with engineered topography
NASA Astrophysics Data System (ADS)
Biofouling is the unwanted accumulation and growth of cells and organisms on clean surfaces. This process occurs readily on unprotected surfaces in both the marine and physiological environments. Surface protection in both systems has typically relied upon toxic materials and biocides. Metallic paints, based on tin and copper, have been extremely successful as antifouling coatings for the hulls of ships by killing the majority of fouling species. Similarly, antibacterial medical coatings incorporate metal-containing compounds such as silver or antibiotics that kill the bacteria. The environmental concerns over the use of toxic paints and biocides in the ocean, the developed antibiotic resistance of bacterial biofilms, and the toxicity concerns with silver suggest the need for non-toxic and non-kill solutions for these systems. The manipulation of surface topography on non-toxic materials at the size scale of the fouling species or bacteria is one approach for the development of alternative coatings. These surfaces would function simply as a physical deterrent of settlement of fouling organisms or a physical obstacle for the adequate formation of a bacterial biofilm without the need to kill the targeted microorganisms. Species-specific topographical designs called engineered topographies have been designed, fabricated and evaluated for potential applications as antifouling marine coatings and material surfaces capable of reducing biofilm formation. Engineered topographies fabricated on the surface of a non-toxic, polydimethylsiloxane elastomer, or silicone, were shown to significantly reduce the attachment of zoospores of a common ship fouling green algae (Ulva) in standard bioassays versus a smooth substrate. Other engineered topographies were effective at significantly deterring the settlement of the cyprids of barnacles (Balanus amphitrite). These results indicate the potential use of engineered topography applied to non-toxic materials as an environmentally friendly coating for antifouling applications in the ocean. In addition, a biomaterial-grade silicone modified with a tailored engineered topography significantly inhibited the bacterial biofilm growth from Staphylococcus aureus for up to 14 days exposure without the use of bactericidal agents. Mature biofilms were present on equivalently exposed smooth silicone surfaces. Engineered surface topographies present a promising means of blocking biofilm development on medical surfaces and reducing the rate of related infections.
Schumacher, James Frederick
Physicochemical changes of microbe and solid surface properties during biofilm formation
NASA Astrophysics Data System (ADS)
Cell immobilization is a promising biotechnology process. For example, entrapment of bacteria cells on synthetic polymeric matrices such as biocarriers is widely used for wastewater treatment because they have strong mechanical strength and durability in contrast to natural polymers. This method is based on the formation of biofilm on the surface of the used carriers and combines two different processes; attached and suspended biomass in a hybrid system. Previous studies have shown that immobilized cell systems have the potential to degrade toxic chemicals faster than conventional wastewater treatment systems because high densities of specialized microorganisms are used in immobilized cell systems. The present study elucidates the surface charge and properties of activated sludge and their role in the formation of biofilm. This information can be used for the optimization of the formation of biofilms as well as for the study of the transport of microorganisms in different environments. The two types of biocarriers that were used in this study are polyvinyl alcohol (PVA)-gel beads and Moving Bed Biofilm Reactor (MBBR) carriers. The sludge samples that were investigated were taken from the aeration tank of the wastewater treatment plant of University of Patras (Greece). Measurements of the surface charge of the sludge, the biocarriers and the formed biofilm, were performed using potentiometric mass titrations with different kinds of electrolytes (e.g. NaCl, NaNO3) and at pH ranging from 3 to 11. The determination of pzc and surface charge of activated sludge and biocarriers is significant, because it can provide new valuable informations about the interaction mechanisms and the formation of biofilms. In each case, the point of zero charge (pzc) was identified as the common intersection point of the potentiometric curve of the blank solution of the electrolyte with the corresponding curves of each material. The pzc value for the biofilm was 6.1 to 6.7 and 6.6 to 6.9 for PVA gel and MBBR, respectively. These values differ both from the pzc values found for PVA biocarriers (pzc = 9.4; no pzc value was obtained for MBBR as expected based on its hydrophobic nature and the absence of surface groups with acid-base behavior) and the pzc value of activated sludge (activated sludge mixed liquor: pzc = 8.0 to 8.2, solid activated sludge: pzc = 7.2 to 7.3). These results lead us to the conclusion that the formed biofilms have different acid-base behavior and properties in relation to the activated sludge and the biocarriers. This fact is in accordance to previous studies, where biofilm-associated cells can be differentiated from their suspended counterparts due to the generation of an extracellular polymeric substance (EPS) matrix. One other possible explanation is that the complicated processes of the biofilm formation can alter the distribution of different cells in the sludge compared with the cell distribution in the suspended unsupported sludge.
Sfaelou, Stavroula; Vakros, John; Manariotis, Ioannis D.; Karapanagioti, Hrissi K.
D-amino acids trigger biofilm disassembly.
Bacteria form communities known as biofilms, which disassemble over time. In our studies outlined here, we found that, before biofilm disassembly, Bacillus subtilis produced a factor that prevented biofilm formation and could break down existing biofilms. The factor was shown to be a mixture of D-leucine, D-methionine, D-tyrosine, and D-tryptophan that could act at nanomolar concentrations. D-amino acid treatment caused the release of amyloid fibers that linked cells in the biofilm together. Mutants able to form biofilms in the presence of D-amino acids contained alterations in a protein (YqxM) required for the formation and anchoring of the fibers to the cell. D-amino acids also prevented biofilm formation by Staphylococcus aureus and Pseudomonas aeruginosa. D-amino acids are produced by many bacteria and, thus, may be a widespread signal for biofilm disassembly. PMID:20431016
Kolodkin-Gal, Ilana; Romero, Diego; Cao, Shugeng; Clardy, Jon; Kolter, Roberto; Losick, Richard
Antibacterial activity of Thymoquinone, an active principle of Nigella sativa and its potency to prevent bacterial biofilm formation
Background Thymoquinone is an active principle of Nigella sativa seed known as "Habbah Al-Sauda" in Arabic countries and "Sinouj" in Tunisia. Bacterial biofilms tend to exhibit significant tolerance to antimicrobials drugs during infections. Methods The antibacterial activity of Thymoquinone (TQ) and its biofilm inhibition potencies were investigated on 11 human pathogenic bacteria. The growth and development of the biofilm were assessed using the crystal violet (CV) and the 2, 3-bis [2-methyloxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide (XTT) reduction assay. Results TQ exhibited a significant bactericidal activity against the majority of the tested bacteria (MICs values ranged from 8 to 32 ?g/ml) especially Gram positive cocci (Staphylococcus aureus ATCC 25923 and Staphylococcus epidermidis CIP 106510). Crystal violet assay demonstrated that the minimum biofilm inhibition concentration (BIC50) was reached with 22 and 60 ?g/ml for Staphylococcus aureus ATCC 25923 and Staphylococcus epidermidis CIP 106510 respectively. In addition our data revealed that cells oxidative activity was influenced by TQ supplementation. In the same way, TQ prevented cell adhesion to glass slides surface. Conclusion The ability of TQ to prevent biofilm formation warrants further investigation to explore its use as bioactive substances with antibiofilm potential.
Secondary flow as a mechanism for the formation of biofilm streamers.
In most environments, such as natural aquatic systems, bacteria are found predominantly in self-organized sessile communities known as biofilms. In the presence of a significant flow, mature multispecies biofilms often develop into long filamentous structures called streamers, which can greatly influence ecosystem processes by increasing transient storage and cycling of nutrients. However, the interplay between hydrodynamic stresses and streamer formation is still unclear. Here, we show that suspended thread-like biofilms steadily develop in zigzag microchannels with different radii of curvature. Numerical simulations of a low-Reynolds-number flow around these corners indicate the presence of a secondary vortical motion whose intensity is related to the bending angle of the turn. We demonstrate that the formation of streamers is directly proportional to the intensity of the secondary flow around the corners. In addition, we show that a model of an elastic filament in a two-dimensional corner flow is able to explain how the streamers can cross fluid streamlines and connect corners located at the opposite sides of the channel. PMID:21402020
Rusconi, Roberto; Lecuyer, Sigolene; Autrusson, Nicolas; Guglielmini, Laura; Stone, Howard A
Secondary Flow as a Mechanism for the Formation of Biofilm Streamers
In most environments, such as natural aquatic systems, bacteria are found predominantly in self-organized sessile communities known as biofilms. In the presence of a significant flow, mature multispecies biofilms often develop into long filamentous structures called streamers, which can greatly influence ecosystem processes by increasing transient storage and cycling of nutrients. However, the interplay between hydrodynamic stresses and streamer formation is still unclear. Here, we show that suspended thread-like biofilms steadily develop in zigzag microchannels with different radii of curvature. Numerical simulations of a low-Reynolds-number flow around these corners indicate the presence of a secondary vortical motion whose intensity is related to the bending angle of the turn. We demonstrate that the formation of streamers is directly proportional to the intensity of the secondary flow around the corners. In addition, we show that a model of an elastic filament in a two-dimensional corner flow is able to explain how the streamers can cross fluid streamlines and connect corners located at the opposite sides of the channel.
Rusconi, Roberto; Lecuyer, Sigolene; Autrusson, Nicolas; Guglielmini, Laura; Stone, Howard A.
Medicinal plants extracts affect virulence factors expression and biofilm formation by the uropathogenic Escherichia coli.
Medicinal plants are an important source for the therapeutic remedies of various diseases including urinary tract infections. This prompted us to perform research in this area. We decided to focus on medicinal plants species used in urinary tract infections prevention. The aim of our study was to determine the influence of Betula pendula, Equisetum arvense, Herniaria glabra, Galium odoratum, Urtica dioica, and Vaccinium vitis-idaea extracts on bacterial survival and virulence factors involved in tissue colonization and biofilm formation of the uropathogenic Escherichia coli rods. Qualitative and quantitative analysis of plant extracts were performed. Antimicrobial assay relied on the estimation of the colony forming unit number. Hydrophobicity of cells was established by salt aggregation test. Using motility agar, the ability of bacteria to move was examined. The erythrocyte hemagglutination test was used for fimbriae P screening. Curli expression was determined using YESCA agar supplemented with congo red. Quantification of biofilm formation was carried out using a microtiter plate assay and a spectrophotometric method. The results of the study indicate significant differences between investigated extracts in their antimicrobial activities. The extracts of H. glabra and V. vitis-idaea showed the highest growth-inhibitory effects (p�<�0.05). Surface hydrophobicity of autoaggregating E. coli strain changed after exposure to all plant extracts, except V. vitis-idaea (p�>�0.05). The B. pendula and U. dioica extracts significantly reduced the motility of the E. coli rods (p�<�0.05). All the extracts exhibited the anti-biofilm activity. PMID:22915095
Wojnicz, Dorota; Kucharska, Alicja Z; Sok�?-??towska, Anna; Kicia, Marta; Tichaczek-Goska, Dorota
Inhibition of Escherichia coli and Proteus mirabilis adhesion and biofilm formation on medical grade silicone surface.
Silicone has been utilized extensively for biomedical devices due to its excellent biocompatibility and biodurability properties. However, its surface is easily colonized by bacteria which will increase the probability of nosocomial infection. In the present work, a hydrophilic antimicrobial carboxymethyl chitosan (CMCS) layer has been grafted on medical grade silicone surface pre-treated with polydopamine (PDA). The increase in hydrophilicity was confirmed from contact angle measurement. Bacterial adhesion tests showed that the PDA-CMCS coating reduced the adhesion of Escherichia coli and Proteus mirabilis by ? 90%. The anti-adhesion property was preserved even after the aging of the functionalized surfaces for 21 days in phosphate-buffered saline (PBS), and also after autoclaving at 121�C for 20 min. Both E. coli and P. mirabilis readily form biofilms on the pristine surface under static and flow conditions but with the PDA-CMCS layer, biofilm formation is inhibited. The flow experiments indicated that it is more difficult to inhibit biofilm formation by the highly motile P. mirabilis as compared to E. coli. No significant cytotoxicity of the modified substrates was observed with 3T3 fibroblasts. PMID:21956834
Wang, Rong; Neoh, Koon Gee; Shi, Zhilong; Kang, En-Tang; Tambyah, Paul Anantharajah; Chiong, Edmund
Inhibition of Candida albicans Biofilm Formation by Farnesol, a Quorum-Sensing Molecule
Farnesol is a quorum-sensing molecule that inhibits filamentation in Candida albicans. Both filamentation and quorum sensing are deemed to be important factors in C. albicans biofilm development. Here we examined the effect of farnesol on C. albicans biofilm formation. C. albicans adherent cell populations (after 0, 1, 2, and 4 h of adherence) and preformed biofilms (24 h) were treated
Gordon Ramage; Stephen P. Saville; Brian L. Wickes; J. L. Lopez-Ribot
Biofilm formation as a novel phenotypic feature of adherent-invasive Escherichia coli (AIEC)
BACKGROUND: Crohn's disease (CD) is a high morbidity chronic inflammatory disorder of unknown aetiology. Adherent-invasive Escherichia coli (AIEC) has been recently implicated in the origin and perpetuation of CD. Because bacterial biofilms in the gut mucosa are suspected to play a role in CD and biofilm formation is a feature of certain pathogenic E. coli strains, we compared the biofilm
Margarita Martinez-Medina; Pl�nio Naves; Jorge Blanco; Xavier Aldeguer; Jesus E Blanco; Miguel Blanco; Carmen Ponte; Francisco Soriano; Arlette Darfeuille-Michaud; L Jesus Garcia-Gil
Specific Antibody Can Prevent Fungal Biofilm Formation and This Effect Correlates with Protective Efficacy
One of the most troublesome medical problems today is infection of prosthetic devices with organisms that form polysaccharide biofilms. This combined with increasing antimicrobial drug resistance is making many infectious diseases incurable. Cryptococcus neoformans is a human-pathogenic fungus that has a polysaccharide capsule and can form biofilms in prosthetic medical devices. We developed a system to study cryptococcal biofilm formation
Luis R. Martinez; Arturo Casadevall
Genetic Control of Conventional and Pheromone-Stimulated Biofilm Formation in Candida albicans
Candida albicans can stochastically switch between two phenotypes, white and opaque. Opaque cells are the sexually competent form of C. albicans and therefore undergo efficient polarized growth and mating in the presence of pheromone. In contrast, white cells cannot mate, but are induced � under a specialized set of conditions � to form biofilms in response to pheromone. In this work, we compare the genetic regulation of such �pheromone-stimulated� biofilms with that of �conventional� C. albicans biofilms. In particular, we examined a network of six transcriptional regulators (Bcr1, Brg1, Efg1, Tec1, Ndt80, and Rob1) that mediate conventional biofilm formation for their potential roles in pheromone-stimulated biofilm formation. We show that four of the six transcription factors (Bcr1, Brg1, Rob1, and Tec1) promote formation of both conventional and pheromone-stimulated biofilms, indicating they play general roles in cell cohesion and biofilm development. In addition, we identify the master transcriptional regulator of pheromone-stimulated biofilms as C. albicans Cph1, ortholog of Saccharomyces cerevisiae Ste12. Cph1 regulates mating in C. albicans opaque cells, and here we show that Cph1 is also essential for pheromone-stimulated biofilm formation in white cells. In contrast, Cph1 is dispensable for the formation of conventional biofilms. The regulation of pheromone- stimulated biofilm formation was further investigated by transcriptional profiling and genetic analyses. These studies identified 196 genes that are induced by pheromone signaling during biofilm formation. One of these genes, HGC1, is shown to be required for both conventional and pheromone-stimulated biofilm formation. Taken together, these observations compare and contrast the regulation of conventional and pheromone-stimulated biofilm formation in C. albicans, and demonstrate that Cph1 is required for the latter, but not the former.
Lin, Ching-Hsuan; Kabrawala, Shail; Fox, Emily P.; Nobile, Clarissa J.; Johnson, Alexander D.; Bennett, Richard J.
Nitazoxanide inhibits biofilm formation by Staphylococcus epidermidis by blocking accumulation on surfaces.
Coagulase-negative species of Staphylococcus are often associated with opportunistic hospital-acquired infections that arise from the colonization of indwelling catheters. Here we show that the antiparasitic drug nitazoxanide (NTZ) and its active metabolite, tizoxanide (TIZ), are inhibitory to the growth of Staphylococcus epidermidis and other staphylococci, including methicillin-resistant Staphylococcus aureus strains, under aerobic and microaerobic conditions (MICs, 8 to 16 microg/ml). At sub-MIC levels, NTZ and TIZ also inhibited biofilm production under static conditions by strains of S. epidermidis and Staphylococcus haemolyticus with a 50% inhibitory concentration of approximately 2.5 microg/ml (8 microM). The 5-nitro group was required for biological activity, and a hydrophilic derivative of NTZ (AMIX) also inhibited biofilm formation. NTZ did not disperse the existing biofilm but did block further accumulation. Sub-MICs of NTZ had no effect on primary attachment to surfaces at either 4 or 37 degrees C. The inhibitory action of NTZ and TIZ, but not vancomycin, on biofilm production could be reversed by the addition of zinc salts (2.5 to 40 microM) but not other metals, suggesting that NTZ might target the zinc-dependent accumulation-associated protein (Aap) that mediates accumulation on surfaces. However, neither NTZ nor TIZ formed chelation complexes with zinc salts, based on spectrophotometric and nuclear magnetic resonance analyses, and addition of excess zinc to NTZ-grown bacteria (apo-Aap) did not restore the accumulation phenotype. Our studies suggest that sub-MIC levels of NTZ may affect the assembly or function of cell structures associated with the biofilm phenotype. PMID:20404119
Tchouaffi-Nana, Florence; Ballard, T Eric; Cary, Christine H; Macdonald, Timothy L; Sifri, Costi D; Hoffman, Paul S
Biofilm formation and persistence on abiotic surfaces in the context of food and medical environments.
The biofilm formation on abiotic surfaces in food and medical sectors constitutes a great public health concerns. In fact, biofilms present a persistent source for pathogens, such as Pseudomonas aeruginosa and Staphylococcus aureus, which lead to severe infections such as foodborne and nosocomial infections. Such biofilms are also a source of material deterioration and failure. The environmental conditions, commonly met in food and medical area, seem also to enhance the biofilm formation and their resistance to disinfectant agents. In this regard, this review highlights the effect of environmental conditions on bacterial adhesion and biofilm formation on abiotic surfaces in the context of food and medical environment. It also describes the current and emergent strategies used to study the biofilm formation and its eradication. The mechanisms of biofilm resistance to commercialized disinfectants are also discussed, since this phenomenon remains unclear to date. PMID:24744186
Abdallah, Marwan; Benoliel, Corinne; Drider, Djamel; Dhulster, Pascal; Chihib, Nour-Eddine
Analyses of Spatial Distributions of Sulfate-Reducing Bacteria and Their Activity in Aerobic Wastewater Biofilms
The vertical distribution of sulfate-reducing bacteria (SRB) in aerobic wastewater biofilms grown on rotating disk reactors was investigated by fluorescent in situ hybridization (FISH) with 16S rRNA-targeted oligonucleotide probes. To correlate the vertical distribution of SRB populations with their activity, the microprofiles of O2, H2S, NO2?, NO3?, NH4+, and pH were measured with microelectrodes. In addition, a cross-evaluation of the FISH and microelectrode analyses was performed by comparing them with culture-based approaches and biogeochemical measurements. In situ hybridization revealed that a relatively high abundance of the probe SRB385-stained cells (approximately 109 to 1010 cells per cm3 of biofilm) were evenly distributed throughout the biofilm, even in the oxic surface. The probe SRB660-stained Desulfobulbus spp. were found to be numerically important members of SRB populations (approximately 108 to 109 cells per cm3). The result of microelectrode measurements showed that a high sulfate-reducing activity was found in a narrow anaerobic zone located about 150 to 300 ?m below the biofilm surface and above which an intensive sulfide oxidation zone was found. The biogeochemical measurements showed that elemental sulfur (S0) was an important intermediate of the sulfide reoxidation in such thin wastewater biofilms (approximately 1,500 ?m), which accounted for about 75% of the total S pool in the biofilm. The contribution of an internal Fe-sulfur cycle to the overall sulfur cycle in aerobic wastewater biofilms was insignificant (less than 1%) due to the relatively high sulfate reduction rate.
Okabe, Satoshi; Itoh, Tsukasa; Satoh, Hisashi; Watanabe, Yoshimasa
A Novel Two-Component Response Regulator Links rpf with Biofilm Formation and Virulence of Xanthomonas axonopodis pv. citri
Citrus bacterial canker caused by Xanthomonas axonopodis pv. citri is a serious disease that impacts citrus production worldwide, and X. axonopodis pv. citri is listed as a quarantine pest in certain countries. Biofilm formation is important for the successful development of a pathogenic relationship between various bacteria and their host(s). To understand the mechanisms of biofilm formation by X. axonopodis pv. citri strain XW19, the strain was subjected to transposon mutagenesis. One mutant with a mutation in a two-component response regulator gene that was deficient in biofilm formation on a polystyrene microplate was selected for further study. The protein was designated as BfdR for biofilm formation defective regulator. BfdR from strain XW19 shares 100% amino acid sequence identity with XAC1284 of X. axonopodis pv. citri strain 306 and 30�100% identity with two-component response regulators in various pathogens and environmental microorganisms. The bfdR mutant strain exhibited significantly decreased biofilm formation on the leaf surfaces of Mexican lime compared with the wild type strain. The bfdR mutant was also compromised in its ability to cause canker lesions. The wild-type phenotype was restored by providing pbfdR in trans in the bfdR mutant. Our data indicated that BfdR did not regulate the production of virulence-related extracellular enzymes including amylase, lipase, protease, and lecithinase or the expression of hrpG, rfbC, and katE; however, BfdR controlled the expression of rpfF in XVM2 medium, which mimics cytoplasmic fluids in planta. In conclusion, biofilm formation on leaf surfaces of citrus is important for canker development in X. axonopodis pv. citri XW19. The process is controlled by the two-component response regulator BfdR via regulation of rpfF, which is required for the biosynthesis of a diffusible signal factor.
Huang, Tzu-Pi; Lu, Kuan-Min; Chen, Yu-Hsuan
Extracellular polysaccharides produced by cooling water tower biofilm bacteria and their possible degradation.
The extracellular polymers (EPS) of biofilm bacteria that can cause heat and mass transfer problems in cooling water towers in the petrochemical industry were investigated. In addition, these microorganisms were screened for their ability to grow and degrade their own EPS and the EPS of other species. Twelve bacteria producing the most EPS were isolated from cooling water towers and characterized biochemically by classic and commercial systems. These were species of Pseudomonas, Burkholderia, Aeromonas, Pasteurella, Pantoea, Alcaligenes and Sphingomonas. EPS of these species were obtained by propan-2-ol precipitation and centrifugation from bacterial cultures in media enriched with glucose, sucrose or galactose. EPS yields were of 1.68-4.95 g l(-1). These EPS materials were characterized for total sugar and protein contents. Their total sugar content ranged from 24 to 56% (g sugar g(-1) EPS), and their total protein content ranged from 10 to 28% (g protein g(-1) EPS). The monosaccharide compositions of EPS were determined by HPLC. Generally, these compositions were enriched in galactose and glucose, with lesser amounts of mannose, rhamnose, fructose and arabinose. All bacteria were investigated in terms of EPS degradation. Eight of the bacteria were able to utilize EPS from Burkholderia cepacia, seven of the bacteria were able to utilize EPS from Pseudomonas sp. and Sphingomonas paucimobilis. The greatest viscosity reduction of B. cepacia was obtained with Pseudomonas sp. The results show that the bacteria in this study are able to degrade EPS from biofilms in cooling towers. PMID:18256966
Ceyhan, Nur; Ozdemir, Guven