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Methods and compositions for removing perchlorate and/or nitrate from contaminated material utilizing perc1ace bacteria under anaerobic conditions. Perc1ace is a gram-negative, curved rod, facultative anaerobe which is deposited with the American Type Culture Collection under ATCC No. 202172. Perc1ace may be used as a substitute for anaerobic bacteria which are presently being used in biological systems for removing perchlorate and/or nitrate from water and other contaminated materials, such as soil.
1. A method for removing perchlorate from water, said method comprising the step of contacting water comprising perchlorate at a first concentration with a bacteria under anaerobic conditions for a sufficient time to remove perchlorate from said water to thereby provide water comprising a second concentration of perchlorate which is lower than said first concentration of perchlorate, wherein said bacteria is identified as percl ace bacteria which is deposited at the American Type Culture Collection under ATCC No. 202172.
2. A method for removing perchlorate from water according to claim 1 wherein said first concentration of perchlorate is below about 1,000 and above 18 parts per billion.
3. A method for removing perchlorate from water according to claim 1 wherein said second concentration of perchlorate is below 18 part per billion.
4. A method for removing perchlorate from water according to claim 1 wherein the pH of said water is between about 7.0 and 7.2.
5. A method for removing perchlorate from water according to claim 1 wherein said anaerobic conditions comprise a carbon source selected from the group consisting of organic acids and proteins.
6. A method for removing perchlorate from water according to claim 1 wherein said water further comprises nitrate at a first concentration and wherein said perc1ace bacteria is contacted with said water under anaerobic conditions for a sufficient time to remove nitrate from said water to thereby provide water comprising a second concentration of nitrate which is lower than said first concentration of nitrate.
7. A method for removing perchlorate from water according to claim 1 which includes the additional step of separating said perc1ace bacteria from the water having said second concentration of perchlorate.
8. In a method for removing perchlorate from a material which is contaminated with perchlorate, said method comprising the step of treating said material with a bacteria under anaerobic conditions to remove said perchlorate, the improvement comprising using a bacteria that is identified as perc1ace, which is deposited at the American Tyne Culture Collection under ATCC No. 202172, as the bacteria in said treatment.
9. An improved method according to claim 8 wherein said material is water that is contaminated with perchlorate which is present at a first concentration.
10. An improved method according to claim 9 wherein said water has a pH of between about 7.0 and 7.2.
11. An improved method according to claim 9 wherein said first concentration of perchlorate is below about 1,000 and above 18 parts per billion.
12. An improved method according to claim 8 wherein said material is also contaminated with nitrate.
13. A biologically pure composition comprising a bacteria that is identified as perc1ace which is deposited at the American Type Culture Collection under ATCC No. 202172.
14. A biologically pure composition according to claim 13 which further comprises growth media for said perc1ace bacteria.
15. A composition for use in treating contaminated material under anaerobic conditions to remove perchlorate and/or nitrate therefrom, said composition comprising a bacteria that is identified as perc1ace which is deposited at the American Type Culture Collection under ATCC No. 202172, growth media for said perc1ace bacteria and a carbon source selected from the group consisting of organic acids and proteins.
16. A system wherein perchlorate contaminants are removed from water, said system comprising water which is contaminated with perchlorate, a bacteria that is identified as perc1ace which is deposited at the American Type Culture Collection under ATCC No. 202172, growth media for said perc1ace bacteria and a carbon source selected from the group consisting of organic acids and proteins.
17. A system according to claim 16 wherein said water has a pH of between about 7.0 and 7.2.
18. A system according to claim 17 wherein the temperature of said water is between about 25� C. and about 30� C.
1. Field of the Invention
The present invention relates generally to methods for removing contaminants from water and soil. More particularly, the present invention involves the treatment of groundwater and/or soil with bacteria under anerobic conditions to remove perchlorates and/or nitrates.
2. Description of Related Art
The publications, patents and other reference materials referred to herein to describe the background of the invention and to provide additional detail regarding its practice are hereby incorporated by reference. For convenience, the reference materials are numerically referenced and grouped in the appended bibliography.
Perchlorate (ClO4) is an oxyanion that has been used extensively in the chemical and aerospace industries because it can act as a strong oxidizing agent. Ammonium perchlorate has been used in solid rocket fuel, explosives and pyrotechnics. The mishandling of perchlorate at aerospace-related industrial sites is the likely source of perchlorate that has recently been discovered in surface and ground waters. The evident persistence of perchlorate in the environment and its toxicity to humans at sufficient concentrations has raised concern over drinking water quality standards and possible environmental impacts. Perchlorate is not currently regulated under the Safe Drinking Water Act, although the California Department of Health Services has established an action level of 18 μg L-1 based on an evaluation of toxicity data by the Environmental Protection Agency. Perchlorate has been found in certain drinking water wells in California and Nevada at concentrations that exceed this action level. In response, the California Department of Health Services has advised that water from these wells should not be used as a source of drinking water.
The development of effective and efficient strategies for the remediation of the perchlorate found in groundwater is an area of intense interest (7). Remediation strategies for the removal of perchlorate based on adsorption by activated carbon have not proven to be highly efficient due to rapid saturation of perchlorate adsorption sites. Other advanced procedures for the removal of perchlorate include reverse osmosis and ion exchange. However, these purification processes are very expensive. In addition, the preceding systems are limited due to the fact that once the perchlorate has been removed from groundwater a second disposal strategy is required to deal with the perchlorate that has been collected.
There also has been intense interest in utilizing microorganisms to remove perchlorate from water by biological processes. The degradation of perchlorate utilizing biological systems provides an attractive remediation strategy for a variety of reasons. For example, microorganisms which degrade perchlorate can completely transform perchlorate into an innocuous end-product, namely, chloride (1, 10, 13). Further, biological treatment can be used for the simultaneous treatment of perchlorate and nitrate. Nitrate pollution of groundwater can occur in agricultural regions, and the allowable concentration in potable water sources is strictly regulated. Finally, biological treatment processes can be quite cost effective when compared to the more expensive physical and chemical processes.
The transformation of oxyanions, such as perchlorate and chlorate, occurs in the absence of oxygen as the result of anaerobic respiration. Microbial respiration couples the oxidation of an organic substrate, such as glucose or acetate, to the reduction of a final electron acceptor, usually oxygen. Under anaerobic conditions, the oxidation of organic compounds requires the use of an alternative electron acceptor in place of oxygen, such as nitrate (NO3), manganese (Mn[IV]), iron (Fe[III]), or sulfate (SO4). Bacteria capable of anaerobic respiration are common to soil and sediment environments where anaerobic conditions are prevalent and natural sources of alternate electron acceptors are common. As a highly oxidized compound (+7 oxidation state), perchlorate has a high potential for utilization as an alternate electron acceptor. Perchlorate-reducing bacteria which have been described in scientific literature include Vibrio dechloraticus Cuznesove B-1168 (8, 14), Wolinella succinogenes HAP-1 (21), and a proteobacteria (strain GR-1) described by Rikken et al. (13). Bacteria capable of reducing chlorate are also known. Examples include Ideonella dechloratans (11) and an Acinetobacter sp. (16).
The proposed pathway of anaerobic perchlorate reduction is as follows (13). ##STR1##
Several patents have described microbial-mediated reduction of perchlorate as a means of removing perchlorate from industrial waste water (2, 8, 23). A United States patent issued to Yakovlev et al. (23) describes the use of unaerated sewage sludge for the treatment of certain oxygen-containing inorganic chlorine and metal compounds, including perchlorate, chlorate and chromate. Domestic sewage sludge is mixed with contaminated wastewater and placed in a large tank. In the absence of aeration, microbial utilization of organic material within the sludge will rapidly deplete the available oxygen. Under anaerobic conditions, the reduction of oxygen-containing inorganic compounds occurs with the oxidation of organic compounds. Following the anaerobic phase, a second stage in the process removes the sludge from the water by precipitation. It is important in this process to supply an excess quantity of organic material, as measured by biochemical oxygen demand (BOD), in order to ensure the creation of an anaerobic environment. The patent also states that the BOD must be greater than the amount of oxygen in the form of inorganic oxygen-containing contaminants present in the wastewater.
Several later patents have improved on the above basic process by enhancing the rate and extent of perchlorate reduction and enabling the treatment of higher concentration of perchlorate in the wastewater. Korenkov (8) described a method of reducing perchlorate and chlorate under anaerobic conditions utilizing the bacterium, Vibrio dechloraticans Cuznesove B-1168. This organism is capable of reducing perchlorate and chlorate when grown anaerobically on acetate or ethanol as a carbon source (14). The authors reported reduction rates of perchlorate as high as 70 mg ClO4 - per hour per gram biomass solids (dry weight), and the ability to treat perchlorate concentrations as high as 3 mM (about 300 mg L-1).
Another United States patent (2) describes a process in which contaminated water is added to an anaerobic bioreactor and spiked with a mixed bacterial culture. The bacterial culture contains a specific bacterium, Wolinella succinogenes, which was isolated from domestic sewage sludge for its ability to reduce very high concentrations (>7000 mg/L) of perchlorate (1, 19, 21). High protein organic nutrients were found to support perchlorate reduction. The source of this oxidizable organic matter in the anaerobic bioreactor could be in the form of aged brewers yeast, cottonseed protein or whey powder. A second stage in the process removes nutrients and organic matter to improve the quality of the water for discharge. One advantage of this system is that it does not utilize sewage sludge, and therefore eliminates problems associated with the presence of pathogens. The bacterium isolated was capable of reducing perchlorate concentrations 26-fold greater than in previous reports, and was reported to have a specific perchlorate degradation rate of at least 1492 mg ClO4 - per hour per gram biomass (dry weight). Through the use of a specific isolate and by optimizing nutrient and environment conditions, the anaerobic reactor was capable of greater perchlorate reduction rates than previously reported.
Wallace et al. (19) described the use of an up-flow anaerobic fixed-bed reactor containing a consortium of facultative anaerobic microorganisms including W. succinogenes HAP-1. The 1.2 m (length)�7.6 cm (internal diameter) reactor could reduce 1500 mg per liter perchlorate to less than 100 mg per liter at a rate of 1 g perchlorate per hour per liter. Bioreactor systems have been optimized to treat high perchlorate concentrations (grams per L range), but there is currently little information about treating levels of perchlorate that are less than 1 mg per liter which can occur in subsurface water supplies. The primary objective of a groundwater treatment system is the removal of perchlorate to less than detectable levels.
The focus of the above research has been the remediation of perchlorate at the very high concentrations which are associated with wastewater generated in industrial situations, such as the manufacturing of solid rocket propellant. An additional problem which is of current concern is the presence of perchlorate at moderate to very low concentrations in subsurface water supplies used as a source of drinking water. To be an effective remediation strategy, biological transformation should ensure the reduction of perchlorate to concentrations less than the current action level of 18 mg L-1, and preferably, to below the limit of detection.
Nitrate is another groundwater contaminant that is commonly found in ground waters of agricultural regions. The U.S. Environmental Protection Agency (EPA) has set a limit of 10 mg NO3 - -N per liter for potable water. Nitrate reduction can occur under the same conditions as perchlorate reduction, and the treatment of nitrate contaminated water using anaerobic bioreactor systems is a well established technology (3, 4, 15). Commercialscale bioreactor systems in operation in Europe provide an efficient removal of nitrate and can operate at a high capacity, with some systems able to treat up to 400 m3 per hour (5, 12). It would be desirable to apply similar technologies to the removal of perchlorate from groundwater. The simultaneous removal of perchlorate and nitrate within the same bioreactor would be a desirable feature of any groundwater treatment system.
As is apparent from the above, there is a present and continuing need to develop new processes and systems for removing perchlorate and nitrate from water.
In accordance with the present invention, a gram-negative, curved rod, facultative anaerobe has been isolated which is especially well-suited for use in the removal of perchlorate and/or nitrate from contaminated water and other materials. The bacteria was isolated from activated sewage sludge and has been given the name perc1ace.
The perc1ace bacteria is well suited for use in a wide variety of anaerobic bioreactors which are presently in use for removing contaminants from ground water. As a feature of the present invention, perc1ace may be substituted into any of the existing contaminant removal processes wherein a bacteria is utilized under anaerobic conditions to convert perchlorate to chloride. The perc1ace bacteria can be used in treating contaminated water wherein the initial perchlorate concentration is relatively low. In addition, the perc1ace bacteria is capable of reducing perchlorate levels to less than 0.005 mg per liter. In addition to biologically pure compositions of perc1ace bacteria, the present invention includes methods for using the perc1ace bacteria and the systems in which the bacteria is utilized.
The above discussed and many other features and attendant advantages of the present invention will become better understood by reference to the following detailed description when taken in conjunction with the accompanying drawings.
FIG. 1 depicts results of tests conducted to show production of chloride from perchlorate using perc1ace bacteria. Perchlorate reduction (), chloride formation (▪) and biomass (◯). Each point represents the mean (� standard deviation) of 3 replicates.
FIGS. 2A-2D depict results of tests conducted to show simultaneous reduction of perchlorate and nitrate by perc1ace bacteria. The figures compare perchlorate reduction with () and without (▪) the presence of nitrate and also show nitrate reduction (◯) for the treatments in which nitrate and perchlorate were both present. Each point represents the mean (� standard deviation) of 3 replicates
FIGS. 3A and 3B depict results of tests conducted to show simultaneous reduction of perchlorate and nitrate under saturated-flow conditions. FIG. 3A shows perchlorate and chloride in eluent when 100 mg per liter perchlorate was loaded into the column. FIG. 3B shows perchlorate and nitrate in the eluent when 0.130 mg per liter perchlorate was loaded into the column. The first arrow indicated the point at which 125 mg per liter nitrate was added to the influent. The level of nitrate was then reduced to 20 mg per liter (second arrow).
The perchlorate-reducing isolate, perc1ace, is a gram-negative, curved rod, facultative anaerobe which can grow aerobically on acetate, and in the absence of oxygen, will reduce perchlorate and nitrate. Perclace is capable of growth on perchlorate between 0.1 and 1000 mg per liter, and can reduce perchlorate to levels <0.005 mg per liter. Identification of perc1ace was attempted using 16S-rRNA gene sequence homology (MIDI labs, Newark, Del.).
There was no similarity match strong enough to identify the isolate. Perc1ace had the highest sequence homology (between 90% and 92% similarity) with several members of the beta subclass of the Proteobacteria, indicating that perc1ace can be considered a member of the beta subclass.
Perc1ace joins three other bacteria known to reduce perchlorate (8, 21, 13) as well as two chlorate reducing bacteria (11, 16). Perc1ace shares some similarities to another beta-proteobacteria strain GR-1 isolated in Europe by Rikken et al. (13). Both are gram negative, rod-shaped, facultative anaerobes which can utilize simple organic acids, such as acetate, for the reduction of perchlorate and nitrate under anaerobic conditions. One distinguishing difference is that GR-1 was found to reduce Mn(IV) while perc1ace did not. The Perc1ace anaerobe was deposited with the American Type Culture Collection (Manassas, Va.) on Sep. 17, 1998 and given ATCC No. 202172.
The optimum environmental conditions for perchlorate reduction were investigated using batch studies. The rate of perchlorate reduction was examined over 5� C. intervals between 20 and 40� C. Perc1ace could reduce perchlorate between 20 and 40� C., with optimum activity between 25 and 30� C. Perchlorate reduction was also determined over a pH range between 5.5 to 8.5. Perc1ace was found to reduce perchlorate over a pH range between 6.5 and 8.5 with optimum activity being observed between 7.0 and 7.2.
The ability of perc1ace to utilize electron acceptors other than perchlorate under anaerobic growth conditions was investigated using acetate as the electron donor, and NO3 -, Fe(III), Mn(IV), or SO4 2- as possible electron acceptors. Perc1ace was found to utilize perchlorate and nitrate only, and showed no capacity to reduce Fe(III), Mn(IV), and SO4 2-.
Perc1ace can grow on acetate using three different electron acceptors, namely oxygen, nitrate and perchlorate. We investigated whether growth conditions could alter the ability of perc1ace to reduce perchlorate. Perc1ace was grown on acetate aerobically and anaerobically in the presence of nitrate or perchlorate. After one week the cells from each incubation treatment were washed in sterile water and then transferred into a perchlorate medium. We found that there was no difference in the rate of perchlorate reduction whether perc1ace had previously been grown aerobically, or anaerobically on nitrate compared to cells cultured on perchlorate (data not shown).
Denitrification by perc1ace was evaluated using bubble production, as described by Tiedje (17). A mineral salts media (35 mL) containing acetate (1000 mg L-1) was transferred into 40 mL screw-cap vials. Triplicate vials received either nitrate (1000 mg L-1 as NaNO3) or nitrous oxide (N2 O). In the case of N2 O, the headspace of the vials was evacuated under vacuum and then replaced with H2 O gas. The vials were then inoculated with perc1ace (1% v/v) and sealed air-tight. The vials contained an inverted glass tube to capture nitrogen gas bubbles formed by the reduction of nitrate and nitrous oxide. Within 3 days the media was turbid and gas bubbles had formed within the inner tube, which provides a presumptive positive for the production of nitrogen gas from the reduction of nitrate and nitrous oxide. As a control, a second set of vials contained perchlorate as the electron acceptor. Turbidity was also evident in the perchlorate vials, but no gas bubbles were formed.
A variety of carbon sources were examined to determine the range of compounds that could be utilized as electron donors in the promotion of perchlorate reduction (Table 1). Perc1ace utilized a variety of commercially available protein sources, including TSB (Difco, Detroit, Mich.) and nutrient broth (Difco), and could also utilize certain organic acids. Sugars and alcohols were not utilized.
TABLE 1______________________________________Carbon source utilizationCarbon source Perchlorate reduction No perchlorate reduction______________________________________Carboxylic acids Acetate Citrate Fumarate Formate Propionate SuccinateSugars Glucose Lactose Sucrose Fructose StarchProtein sources Casamino acids Nutrient broth Peptone Tryptic Soy Broth Yeast extractAlcohols Methanol Ethanol______________________________________
The production of chloride from perchlorate reduction is shown in FIG. 1. Perc1ace reduced 580 mg per liter ClO4 - over a 72 hour period with the contaminant release of chloride, and the production of biomass, as indicated by an increase in optical density. After 72 hours, the amount of chloride produced represented between 97 and 105% of what had been added as perchlorate, indicating that perchlorate was completely transformed.
Perc1ace is capable of reducing both perchlorate and nitrate under similar growth conditions. FIG. 2 explores the effect of nitrate on perchlorate reduction by holding nitrate levels at 1 mM (62 mg per liter) and varying perchlorate between approximately 1 mM to 0.001 mM. With all treatment combinations it is evident that when perchlorate and nitrate are present together, nitrate was reduced at a faster rate than perchlorate. However, both nitrate and perchlorate were reduced within 48 hours. At almost equal molar concentrations of nitrate and perchlorate (FIG. 2A), there was little difference between perchlorate reduction in the presence or absence of nitrate. When perchlorate levels were reduced 10-, 100-, and 1000-fold compared to nitrate, the presence of nitrate did decrease the rate of perchlorate reduction, as compared to perchlorate reduction in the absence of nitrate. For example, the reduction of 0.089 mg per liter perchlorate reduction (FIG. 2D) to <0.005 mg per liter required less than 24 hours in the absence of nitrate but required 36 hours in the presence of 62 mg per liter of nitrate. It is interesting to note that perchlorate also affected the rate of nitrate reduction. At equal molar concentrations of nitrate and perchlorate (FIG. 2A), nitrate reduction required 48 hours. When the perchlorate concentration was reduced to 0.089 mg per liter (FIG. 2D), nitrate reduction required less than 24 hours. We also monitored for the presence of the nitrate reduction intermediate, nitrate (NO2 -), and found that there was no detectable levels of NO2 - produced during NO3 - reduction.
It should be noted that the complete transformation of ClO.sub.- to Cl- was not affected by the presence of nitrate. Chloride production was monitored when 122 mg per liter perchlorate was reduced (FIG. 2A) and revealed the complete transformation of ClO4 - to cl- in the presence and absence of nitrate.
A column study was used to determine if the simultaneous reduction of perchlorate and nitrate would be maintained under saturated-flow conditions. The sand-packed column was first inoculated with perc1ace and then treated with a high perchlorate concentration to establish a perchlorate-reducing population. FIG. 3A shows that within one day, perchlorate concentrations were reduced from 100 mg per liter in the influent to <1 mg per liter in the eluent. Chloride determination within the eluent revealed complete transformation of perchlorate to chloride. The influent perchlorate concentration was then reduced to 0.130 mg per liter. After approximately 6 days (51 pore 25 volumes), 125 mg per liter nitrate was added to the influent for the next 40 pore volumes. Following the addition of nitrate, perchlorate levels in the eluent initially increased to 0.099 mg per liter. But within 5 pore volumes, perchlorate levels had declined to <0.005 mg per liter, and remained below detection for the next 35 pore volumes. Eluent nitrate concentration ranged from 1.0 to 3.5 mg per liter in the first 5 pore volumes, and then decreased to <0.5 mg per liter. Nitrite was detected in the eluent and ranged from 2 to 43 mg per liter for the first 16 pore volumes, and then was reduced to <0.5 mg per liter for the remainder of the study (data not shown). At pore volume 91, the nitrate level in the influent was reduced from 125 mg per liter to 20 mg per liter for the next 38 pore volumes. The reduction in nitrate level was followed by a period of perchlorate breakthrough. Perchlorate levels between 0.009 and 0.026 mg per liter were present in the eluent for 7 pore volumes, and then decreased to <0.005 mg per liter for the remaining 31 pore volumes. Although two samples (pore volumes 110 and 120) showed perchlorate levels of 0.0045 mg per liter. Nitrate and nitrite in the eluent remained <0.5 mg per liter.
The perc1ace bacteria may be used in place of any of the other bacteria which have been used to treat water and other materials to remove perchlorate and/or nitrate contaminants. For example, perc1ace can be used in the systems described in References 1-5, 8, 12-15, 19, 23. The term "biologically pure" when used herein refers to the perc1ace bacteria once it has been separated from its naturally occurring surroundings, i.e. sewage sludge. The perc1ace bacteria may be grown and cultured using conventional bacterial culture media which are well-known in the art. Exemplary growth media and conditions are as follows: perc1ace bacteria can be grown in a mineral salts media with acetate as the sole carbon source. The mineral salts media used contained K2 HPO4 (225 mg L-1), KH2 PO4 (225 mg L-1), (NH4)2 SO4 (225 mg L-1), MgSO4 7H2 O (50 mg L-1), CaCO3 (5mg L-1), FeCl2 (5 mg L-1), plus a mixture of trace metals. The acetate can be replaced with other organic acids, including fumarate, propionate and succinate, or the acetate can be replaced with protein sources, such as casamino acids, peptone or yeast extract. Perchlorate reduction requires the absence of oxygen, as well as moderate temperature (between 20 and 40� C.) and pH (between 6.5 and 8.5) conditions.
Perc1ace is especially well-suited for removing low levels perchlorate from water where the initial or first concentration of perchlorate is below about 1000 μg L-1, but above 18 μg L-1. When perc1ace is used to treat water having such relatively low contamination levels, the resulting treated water will have a final or second concentration of perchlorate which is below the mandated limit of 18 μg L-1.
Details regarding the isolation and examination of perc1ace are as follows:
Isolation of the perc1ace bacterium. Enrichment for a perchlorate reducing bacterial isolate began with the transfer of 10 mL of activated sewage sludge (Water Quality Control Plant, Riverside, Calif.) into a 125 mL Erlenmeyer flask containing 125 mL mineral salts medium with 1000 mg per liter acetate and 500 mg per liter perchlorate (NaClO4, Aldrich, Milwaukee, Wis.). The headspace of the Erlenmeyer flasks were purged with nitrogen gas and sealed air-tight with screw-cap stoppers. The mineral salts medium, FTW, modified from Losi and Frankenberger (9), contained K2 HPO4 (225 mg per liter), KH2 PO4 (225 mg per liter), (NH4)2 SO4 (225 mg per liter), MgSO4 7H2 O (50 mg per liter), CaCO3 (5 mg per liter), FeCl2 (5 mg per liter), plus a mixture of trace metals. Perchlorate levels within the flask were monitored using a perchlorate specific electrode (described below) and the loss of perchlorate was detected after more than 1 month. The perchlorate-reducing enrichment culture was then transferred to fresh medium, and has been maintained by monthly transfers to fresh medium. The perchlorate-reducing bacterium was isolated by selective plating using agar-hardened FTW containing 500 mg per liter perchlorate and 1000 mg per liter acetate. The plates were streaked with the perchlorate-reducing enrichment culture and incubated in a BBL GasPack anaerobic jar (Becton Dickinson, Cockeysville, Md.). After two weeks incubation, the dominant colony type was transferred twice more to fresh plates to ensure that a single species was isolated. The isolate was named perc1ace and was deposited with the American Type Culture Collection (ATCC) on Sep. 17, 1998 as deposit No. 202172.
The ability of perc1ace to utilize a variety of electron acceptors other than perchlorate was examined. With acetate as the electron donor, growth of the bacteria under anaerobic conditions was determined in 125 mL of FTW containing 1000 mg per liter acetate with NO3 -, Fe(III), Mn(IV), or SO4 2- as electron acceptors, and a 1% (v/v) inoculum of perchlorate-grown perc1ace. Five mM of NO3 - (as NaNO3) or SO4 2- (as Na2 SO4) were used. The Fe(III) and Mn(IV) were prepared as oxide slurries following the procedures outlined by Ghiorse (6), and 5 mM suspensions in FTW were prepared based on the dry weight of the slurries assuming that Fe(III) or Mn(I) were dominant. The ability of the bacteria to utilize each electron acceptor was evaluated by biomass production (turbidity) as well as color change indicators. SO2- reduction was indicated by the formation of a black precipitate due to a reaction of sulfide with the iron in FTW. Iron reduction was indicated by the conversion of red-brown Fe(III) oxide to a black precipitate, while the reduction of dark brown Mn(IV) formed a white precipitate. Nitrate loss was determined by ion chromatography, as described below.
Perchlorate, nitrate, nitrite, and chloride analysis. Perchlorate concentrations were determined using an ion selective probe (model 93-81, Orion Research, Boston, Mass.) for perchlorate concentrations between 1 and 1000 mg per liter. Detection of perchlorate to 0.005 mg per liter was performed by ion chromatography (IC) (Dionex, Sunnyvale, Calif.). Initially an IonPac AS5 column was used, but we later switched to an LonPac AS11 column and followed the method described in Dionex application note #121 and by Wirt et al. (22). We used a 100 mM NaOH eluent at a flow rate of 1 mL per minute with conductivity detection. Eluent suppression was performed using ASRS-II (4 mm) operated at 300 mA with water as the regenerate (10 mL per minute). The sampling loop was 0.740 mL.
Nitrate and nitrite concentration to <0.5 mg per liter were determined by ion chromatography using an IonPac AS14 column following the standard procedure provided by Dionex. Chloride was determined using a HBI digital chloridometer.
Perchlorate-reduction batch studies. Batch studies were performed using 50 mL Erlenmeyer flasks sealed with rubber stoppers (size #2). The flasks contained 50 mL of FTW with 1000 mg per liter acetate and the appropriate amount of perchlorate, depending on the study. The inoculum was prepared by washing perchlorate grown perc1ace in sterile water and then adding 1% (v/v) aliquot.
A batch study was used to demonstrate perchlorate reduction, chloride formation, and biomass production for perc1ace. The formation of chloride and the production of biomass from the reduction of 580 mg per liter perchlorate was monitored after 12, 24, 36, 48, 56, 65, and 72 hours. A high concentration of perchlorate was used such that chloride formation could be detected above background levels. At each sampling time, 3 flasks were sacrificed, and samples for perchlorate and chloride determination were centrifuged and then filtered (Supor-450 membranes, 0.45 μM, Gelman Sciences, Ann Arbor, Mich.). Perchlorate loss and chloride formation were monitored using the Orion electrode and the chloridometer, respectively, while biomass production was determined by an increase in optical density (OD600).
Groundwater contaminated with perchlorate often contains appreciable amounts of nitrate. The effect of nitrate on perchlorate reduction by perc1ace was examined in a series of batch experiments in which the NO3 - concentration was maintained at 62 mg per liter (1.0 mM, added as NaNO3) while ClO4 - levels were 122, 12.0, 0.92, and 0.089 mg per liter (1.2, 0.12, 0.01, and 0.001 mM). Perchlorate reduction was monitored in the presence and absence of nitrate, with samples usually taken every 12 hours over a 48 hour period. The perchlorate and/or nitrate in 50 mL of FTW with 1000 mg per liter acetate and 1% (v/v) inoculum of perchlorate-grown perc1ace was incubated in 50 mL Erlenmeyer flasks sealed with rubber stoppers. In the experiments with perchlorate at 122 and 12.0 mg per liter, perchlorate removal to <1 mg per liter was monitored using the perchlorate selective electrode. Ion chromatography was used to monitor the loss of perchlorate in the remaining experiments. In the case of 0.92% mg per liter perchlorate, the FTW medium was found to interfere with perchlorate analysis and limited the detection of perchlorate to 0.1 mg per liter. In the case of 0.089 mg per liter perchlorate, a change was made to a less concentrated mineral salts formulation. The new mineral salts medium, referred to as BMS, was composed of NaH2 PO4 �H2 O (36 mg per liter), Na2 HPO4 (104 mg per liter), NH4 Cl (60 mg per liter), MgSO4 �7H2 O (10 mg per liter), CaCl2 �2H2 O (10 mg per liter), and FeCl2 �4H2 O (2 mg per liter). This formulation provided nutrients essential to maintaining perchlorate reduction but interfered less with perchlorate detection, and perchlorate loss could be detected to 0.005 mg per liter. Perchlorate and/or nitrate in BMS (40 mL) with 300 mg per liter acetate was incubated in 40 mL EPA vials (Fisher Scientific) sealed with teflon-lined screw-caps.
Perchlorate-reduction column study. A glass column (2.8 cm internal diameter and 14 cm length) packed with sterilized, oven-dried sand (Type #30, Ogleby Norton Industrial Sands, San Juan Capistrano, Calif., 40-70 mesh size was used). The pore volume within the column was 36 mL, and a 3 hour retention time was maintained using a 0.2 mL per minute flow rate (model P-3 peristaltic pump, Pharmacia, Piscataway, N.J.). The column was saturated for 5 days from the bottom-up with a 5 mM NaCl solution, and then inoculated by loading the column for 24 hours with a suspension of perc1ace. The cell suspension was prepared using perchlorate-grown cells washed and suspended in sterile FTW to a final optical density (OD600) of 0.05. The column was then loaded with a solution of 100 mg per liter perchlorate and 1000 mg per liter acetate in FTW for 6 days. The high perchlorate influent was used to establish a viable and active perchlorate-reducing population within the column. Perchlorate concentration in the influent and eluent were determined daily using the perchlorate specific electrode, and the transformation of perchlorate was determined by monitoring chloride levels in the eluent. Following the establishment of a perchlorate-reducing population, the column study then proceeded in two stages. The objective of the first stage was to show perchlorate reduction using a level of perchlorate representative of contaminated groundwater. The column was loaded with 0.130 mg per liter perchlorate and 300 mg per liter acetate in BMS medium for 6 days. The influent medium had been autoclaved, and was purged constantly under a nitrogen stream in order to maintain low oxygen concentrations. In the second stage of the column study, nitrate was added to the influent in order to determine if perc1ace could maintain perchlorate reduction in the presence of an alternate electron acceptor. Two different levels of nitrate were used to represent groundwater contaminated with a relatively high level of nitrate (125 mg per liter) and a more moderate level of nitrate (20 mg per liter). The column influent was 0.130 mg per liter perchlorate with 300 mg per liter acetate plus 125 mg per liter nitrate for five days, followed by 20 mg per liter nitrate for five days. The column eluent was collected using a fraction collector, and selected fractions were filtered and then analyzed for perchlorate and nitrate using ion chromatography.
Having thus described exemplary embodiments of the present invention, it should be noted by those skilled in the art that the within disclosures are exemplary only and that various other alternatives, adaptations, and modifications may be made within the scope of the present invention. Accordingly, the present invention is not limited to the specific embodiments as illustrated herein, but is only limited by the following claims.
1. Attaway, H., and M. Smith. 1993. Reduction of perchlorate by an anaerobic enrichment culture. J. Ind. Microbiol. 12:408-412.
2. Attaway, H., and M. Smith. 1994. Propellant wastewater treatment process. U.S. Pat. No. 5,302,285.
3. Chen, S-D., C.-Y. Chen, Y.-C. Shen, C.-M. Chiu and H.-J. Cheng. 1996. Treatment of high-strength nitrate wastewater by biological methods operational characteristics study. Wat. Sci. Tech 34:269-276.
4. Csikor, Zs., L. Czako, P. Mihaltz, and J. Hollo. 1996. Complete nitrogen removal from waste and drinking water in a fluidized-bed reactor. Food Science and Technology International 2:165-171.
5. Gayle, B. P., G. D. Boardman, J. H. Sherrard, and R. E. Benoit. 1989. Biological denitrification of water. J. Environ. Engineering 115:930-943.
6. Ghiorse, W. C. 1994. Iron and manganese oxidation and reduction In Methods of Soil Analysis, Part 2--Microbiological and Biochemical Properties. Soil Science Society of America, Book Series 5. SSSA, Madison, Wis. pp. 1079-1096.
7. Herman, D. C., and W. T. Frankenberger, Jr. 1998. Microbial-mediated reduction of perchlorate in groundwater. J. Environ. Qual. 27:750-754.
8. Korenkov, V. N., V. I. Romanenko, S. I. Kuznetsov, and J. J. V. Voronov. 1976. Process for purification of industrial waste waters from perchlorates and chlorates. U.S. Pat. No. 3,943,055.
9. Losi, M. E., and W. T. Frankenberger, Jr. 1997. Reduction of selenium oxyanions by Enterobacter cloacae SLD1a--a: isolation and growth of the bacterium and its expulsion of selenium particles. Appl. Environ. Microb. 63:3079-3084.
10. Malmqvist, A., T. Welander, and L. Gunnarsson. 1991. Anaerobic growth of microorganisms with chlorate as an electron acceptor. Appl. Environ. Microbiol. 57:2229-2232.
11. Malmqvist, A., T. Welander, E. Moore, A. Ternstrom, G. Molin, and I.-M. Stenstrom. 1994. Ideonella dechloratans gen. nov., sp. nov., a new bacterium capable of growing anaerobically with chlorate as an electron acceptor. System. Appl. Microbiol. 17:58-64.
12. Mateju, V., S. Cizinska, J. Krejci, and T. Janoch. 1992. Biological water denitrification--a review. Enzyme Microbiol. Technol. 14:170-183.
13. Rikken, G. B., A. G. M. Kroon, C. G. van Ginkel. 1996. Transformation of (per)chlorate into chloride by a newly isolated bacterium: reduction and dismutation. Appl. Microbiol. Biotechnol. 45:420-426.
14. Romanenko, V. I., V. N. Korenkov, and S. I. Kuznetsov. 1976. Bacterial decomposition of ammonium perchlorate. Mikrobiologiya 45:204-209.
15. Semon, J., T. Sadick, D. Palumbo, M. Santoro, and P. Keenan. 1997. Biological upflow fluidized bed denitrification reactor demonstration project--Stamford, Conn., USA. Wat. Sci. Tech. 36:139-146.
16. Stepanyuk, V. V., Smirnova, G. F., Klyushnikova, T. M., Kanyuk, N. I., Panchenko, L. P., Nogina, T. M., and Prima, V. I., 1992. New species of the Acinetobacter genus, Acinetobacter thermotoleranticus sp. Nov. Mikrobiologiya 61:347-356.
17. Tiedje, J. M. 1994. Denitrifiers. In Methods of Soil Analysis, Part 2--Microbiological and Biochemical Properties. Soil Science Society of America, Book Series 5. SSSA, Madison, Wis. pp. 245-267.
18. van Ginkel, C. G., G. B. Rikken, A. G. M. Kroon, and S. W. M. Kengen. 1996. Purification and characterization of chlorite dismutase: a novel oxygen-generating enzyme. Arch Microbiol. 166:321-326.
19. Wallace, W., S. Beschear, D. Williams, S. Hospadar, and M. Owens. 1998. Perchlorate reduction by a mixed culture in an up-flow anaerobic fixed bed reactor. J. Ind. Microbiol. Bioteck. 20:126-131.
20. Wallace, W., S. Beschear, S. Wyatt, and S. Baxley. 1995. Purification of the chlorite reductase from Wolinella succinogenes HAP-1, Abstract O-53-, p. 377. In Abstracts of the 95th General Meeting of the American Society for Microbiology, American Society for Microbiology, Washington, D.C.
21. Wallace, W., T. Ward, A. Breen, and H. Attaway. 1996. Identification of an anaerobic bacterium which reduces perchlorate and chlorate as Wolinella succinogenes. J. Ind. Microbiol. 16:68-72.
22. Wirt, K., M. Laikhtman, J. Rohrer, P. E. Jackson. 1998. Low-level perchlorate analysis in drinking water and ground water by ion chromatography. American Environmental Laboratory 10(3):4-5.
23. Yakovlev, S. V., J. V. Voronov, V. N. Korenkov, A. B. Nevsky, V. A. Bobrikova, T. A. Karjurkhina, I. N. Churbanova, and J. M. Laskov, J. M. 1973. Method for biochemical treatment of industrial waste water. U.S. Pat. No. 3,755,156.
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SACRAMENTO, Calif. (AP) — California bank to pay $4 million to Democratic politicians defrauded by campaign treasurer.
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Man arrested; deputies say he aimed banana at them
GRAND JUNCTION, Colo. (AP) — A man is facing a felony menacing charge after two western Colorado sheriff's deputies say he pointed a banana at them and they thought it was a gun. Full Story
Garcia Marquez family explains archive decision
MEXICO CITY (AP) — The family of Nobel laureate Gabriel Garcia Marquez says the late Colombian novelist's archive was offered only to the University of Texas, which will be the materials' repository. Full Story
(Adrian Sanchez-Gonzalez/Getty Images)
Bette Midler to Ariana Grande: Clean up your act
The veteran entertainer complains in an interview that young singers rely too much on selling sex instead of talent.

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Explorer Map 157 Marlborough and Savernake Forest
With its exceptional level of detail and clarity, the Explorer series of OS Maps is a favourite of walkers, climbers, mountain bikers, outdoor professionals and anyone else who needs to know exactly where they are! The 1:25,000 scale allows plenty of detail, and the squares of the National Grid allow every bit of Great Britain to be given its own unique reference number.Masses of features are displayed, including areas of interest, picnic spots, campsites and National Trails, along with National Park and Rights of Way information for England and Wales.
FREE STANDARD UK DELIVERY 20 - 23 November 2014
Product Features
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Explorer Map OL7 The English Lakes - South Eastern Area
Explorer Map 151 Cardiff and Bridgend
Explorer Map OL17 Snowdon - Conwy Valley
Explorer Map OL6 The English Lakes - South Western Area
Explorer Map OL24 The Peak District - White Peak Area
Explorer Map 206 Edge Hill and Fenny Compton
Landranger Map 182 Weston-Super-Mare
Explorer Map OL1 The Peak District - Dark Park Area
Explorer Map OL4 The English Lakes - North Western Area
Explorer Map OL5 The English Lakes - North Eastern Area


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Latest News
Joe Giudice: I'll Worry About Deportation 'When The Time Comes'
With everything on his plate these days, getting kicked out of the country is the last thing on Joe Giudice's mind.
In a preview of the second part of Watch Wha… READ MORE
Kim DePaola: Teresa & Joe Giudice Believe 'The Show Brought This Out'...
Teresa Giudice's bestie Kim DePaola, speaking about the reality star's upcoming 15-month prison sentence in her fraud conviction, said that the "underl… READ MORE
Former Bankruptcy Attorney For Giudice Family Defends Teresa's Claims...
Teresa Giudice told viewers that she "didn't understand" that her plea deal would mean prison time during her Watch What Happens Live interview, but a sign… READ MORE
Teresa Giudice Admits, 'I Wish I Could Do Everything Differently'
It goes without saying, but Teresa Giudice is in a very bad place these days.
The Real Housewives of New Jersey star, set to surrender herself to authorities … READ MORE
Tom Cruise's Former Bodyguard, Sean Ringgold, Cleared Of Rape Charges
A year after a court indicted former celebrity bodyguard and soap star Sean Ringgold for rape and sexual assault, all charges against him have been dismiss… READ MORE
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"It is not a scandal; it is a sex crime," Jennifer Lawrence says of the hundreds of naked pics that hackers leaked of her and dozens of other celebs last month, … READ MORE
NYPD Probes Actor Stephen Collins In Sex Crimes Toward 3 Girls: Report...
Actor Stephen Collins, who played a family man and minister on the CW TV hit 7th Heaven, admitted to molesting at least three girls in a 2012 therapy session w… READ MORE
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Teresa Giudice's sentence to 15 months in prison "was not [an] unexpected" outcome for the Real Housewives of New Jersey star in her sentencing hear… READ MORE
Giudices' Lawyer Blames Bravo, Death Of Joe's Dad For Couple's Issues...
Real Housewives of New Jersey stars Teresa and Joe Giudice were scrambling down to the wire in the hopes of winning leniency in their fraud sentencing today… READ MORE
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Joe Giudice received a 41-month sentence, while Teresa Giudice was awaiting her sentencing as The Real Housewives of New Jersey stars heard their punishm… READ MORE
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Posted Sep 16, 2012By
The 14th Annual Port of Los Angeles Lobster Festival took place this weekend at the Port O'Call Village in San Pedro.
[insertSlideshowjava xml="http://director-lang-ieng.medianewsgroup.com/images.php?album=1727" api="metro"]
People rally outside the Los Angeles Metropolitan Detention Center Thursday night, November 20, 2014 to hear Presid
Cal State L.A. President William Covino and LA County Supervisor Gloria Molina Thursday, launched a new incubator,
Students at Mira Costa High School returned to school Thursday to added security after the internet threats posted
Members of the San Bernardino County Sheriff and Fire Departments investigate an explosion that occurred on the 330
Thousands of people attended the Manhattan Beach Pier lighting ceremony on Wednesday night. The holiday lights were
The 6th Annual Day of Discovery at the U.S. Coast Guard Base in San Pedro. Local high school students explore caree


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for veterans day
edit: wow went to work had a **** shift, burned myself and didnt get home till midnight. All these thumbs and all these feels made my day so much better.
plus lost front page virginity, and tops.
Top Rated Controversial Best Lowest Rated Newest Per page:Order:
What do you think? Give us your opinion. Anonymous comments allowed.
#48 - savemeelle (11/12/2011) [+] (15 replies)
#38 - ICanThePowerr (11/12/2011) [+] (3 replies)
#16 - Womens Study Major (11/12/2011) [+] (2 replies)
lol that shadow makes no sense
#78 - hunterxwitch (11/12/2011) [-]
this gave me a huge feel ;-; that shadow is so cool!
ps: THANK YOU VETERANS FOR SERVING MY NATION AND COMUNITY
ps: THANK YOU VETERANS FOR SERVING MY NATION AND COMUNITY
#137 - alpachinoderp (11/12/2011) [+] (2 replies)
**alpachinoderp rolls 40** i shall roll dubs for veterans day
#117 - randomawesomfacetr (11/12/2011) [+] (3 replies)**randomawesomfacetr rolled a random image** mfw i see a lonely old man
#32 - samuraicrack (11/12/2011) [+] (3 replies)
WW1 and WW2 were real wars fought to protect people. the vietnam war and the current middle eastern wars are just a waste of time, money and lifes
#45 - chinnerz (11/12/2011) [+] (2 replies)
It's nice to see things like this on the internet. Showing we can all appreciate veterans who have fought for us and ones who still do, whether they're from our country or not.
#13 - erwinrommell (11/12/2011) [+] (1 reply)
Nothing sadder then a brother fighting a brother, or a father killing his own son. **** happened in Eastern Europe - especially the Baltics. Read about it, it is truly a tragedy.
Nothing sadder then a brother fighting a brother, or a father killing his own son. **** happened in Eastern Europe - especially the Baltics. Read about it, it is truly a tragedy.
#36 - icedman (11/12/2011) [-]
#126 - yokiewartooth (11/12/2011) [-]
#1 - puggles (11/11/2011) [+] (2 replies)
something worth think about. i try to give old guys who are annoying me a little more slack, figuring they probably were drafted into the military and gave up a few years of their lives to defend my country. of course they may have just stayed home on a trumped up medical deferment and banged all the military wives that were left at home. either way, thumbs up for them.
#139 - Abra (11/12/2011) [+] (1 reply)
It's not just Veterans day, it's also Remembrance day
#118 - mogwaves (11/12/2011) [+] (2 replies)**mogwaves rolled a random image** This is actuqally what my black ops emblem is, or, was


text:
The Congress has promised citizens of Mangalore that it would not increase house tax and water tariff if it came back to power in the Mangalore City Corporation council.
This is among the key points of party's manifesto for the Corporation elections which will be released on Saturday. Congress's in-charge for MCC election B. Janardhana Poojary said here on Thursday that party workers would highlight these points during the campaign.
Mr. Poojary said in the 15-year rule in the MCC Council the Congress had never increased the house tax and water charges. Soon after coming to power, the BJP increased house tax by 10 times.
Keywords: house tax, Mangalore City Corporation council
Infrastructure trends in poll manifesto
More In: Mangaluru


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|Mon, 10-31-2011 - 9:56pm
Sorry I havent been around much for support and a lot of congratulations. I am still out of town at my parents house and didnt want them to see me posting. I am not temping or using opks this month because I am out of town but according to FF I should ovulate at the end of this week very beginning of next week. I am hoping thay with travel that ovulation may not happen until I get home since Im traveling. I guess we will see.


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You're leaving Wedding Paper Divas to see the Deal of the Day at our parent site, Tiny Prints.
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"I knew I wanted to get magnates for my save the dates, and I'm more of a simple person, so when I saw this design I knew it would be perfect. I am so happy with the way that everything came together. You're able to add your own touch when it comes to the design. I love how when you submit the order, a designer reviews your order and makes suggestions on where to place the text, I took their suggestion, and they turned out great! The delivery is surprisingly fast, the quality of the magnate is great! I was worried at first that the picture would come out grainy! But it turned out very clear! My friends and family love them, because its not something you throw away, they will be able to get it forever!"
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New York Public Library to Host All-Night Scavenger Hunt
In celebration of the New York Public Library's centennial festival weekend, game designer Jane McGonigal has crafted the "Find the Future" scavenger hunt.
500 players will join the "Write All Night" event on May 20th. Inside the Stephen A. Schwarzman Building, they will use laptops and smartphones to find 100 objects from the library's collection of treasures and perform a related-writing challenge.
The video embedded above features a promo clip for the event; it seems to mimic The Da Vinci Code's film trailer. If you want to participate, just answer this question: "In the year 2021, I will become the first person to __________." Submit your answer before 11:59 PM Pacific Time on April 21st.
Here's more from CNN: "When they find writer Charles Dickens' letter opener, which, bizarrely, is made out of his dead cat's taxidermied paw, they get a prompt that asks them to write an open letter to someone they love. When they find a draft copy of the Declaration of Independence, which has scribbling in the margins that suggest Thomas Jefferson was trying get the Founding Fathers to ban slavery, they're asked to write a declaration of their own."
(via The Gothamist)
- Ferguson Public Library Receives $175K in Donations
- The Contemporary Jewish Museum Hosts Exhibit On J. Otto Seibold
- Indies First & Gloria Steinem Get Booked
- The Grolier Club is Hosting the 'Quotations of Chairman Mao' Exhibit


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Quick Pork Dinner Recipes
Looking for more quick pork dinner recipes? Find many more quick pork dinner recipes, healthy quick pork dinner recipes, and quick pork dinner recipe ideas.
Recipe Spotlight
Quick Homemade Sweet-and-Sour Pork
Stir up a homemade sweet-and-sour sauce for this colorful combination of tender pork, crunchy vegetables and tangy pineapple. Serve it over hot rice, chow mein noodles or both. —Eleanor Dunbar, Peoria, IllinoisView Quick Homemade Sweet-and-Sour Pork Recipe


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Find records of Julie Howell
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Save to My Recipes
Peas and Pea Tendrils with Lemon Dressing
Servings: 6
- 1/4 cup olive oil
- 2 tablespoons fresh lemon juice
- 1 teaspoon sugar
- 2 cups shelled fresh peas or one 10-ounce package frozen petite peas, unthawed
- 8 ounces pea tendrils, cut into 4-inch lengths
PREPARATIONView Step-by-Step Directions
Whisk oil, lemon juice, and sugar in small bowl to blend; set dressing aside.
Bring large pot of salted water to boil. Add fresh or frozen peas and cook 3 minutes. Add pea tendrils; cook 1 minute. Drain well. Return vegetables to pot; add oil-lemon dressing and toss to coat. Season with salt and pepper.
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This is a yummy, healthy way to enjoy pizza from my Garden Cuisine cookbook by Paul Wenner. Below are the toppings that I chose however, you can use any of your own pizza favorites. I actually added some Hawaiian sausage to a couple of them for my meat-loving guests Enjoy!
Units: US | Metric
Serving Size: 1 (414 g)
Servings Per Recipe: 4
Make this season merry, bright and manageable with our easy ideas.


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SnapAV is branching out a bit. After years in the custom AV channel, the company is now planning to break into the security market.
The company just announced its WirePath Surveillance line, which includes eight cameras, three digital video recorder (DVR) options, and a free iPhone app (Android is coming soon).
In that camera line, you'll find both dome and bullet styles, offering four different levels of performance. There are also basic and advanced models for the DVR line. Last, but certainly not least, that WirePath Surveillance app allows users to view and control the cameras and DVRs from anywhere in the world. A few of the capabilities include watching real-time video, searching through and playing back recorded video, and taking screen captures.
SnapAV says that most of the WirePath Surveillance products are shipping now. Check the SnapAV website for more info on each of the WirePath products.
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Old Ocean Mold
December 12, 2012|
Fungi in 100 million year-old seafloor sediments could possess novel antibiotics.
December 12, 2012|
Fungi in 100 million year-old seafloor sediments could possess novel antibiotics.
December 10, 2012|
Long-term exposure to antibiotics from agricultural run off may encourage the evolution of soil bacteria that break down and consume the antibacterial agents.
December 6, 2012|
Insulin, long recognized as a primary regulator of blood glucose, is now also understood to play key roles in neuroplasticity, neuromodulation, and neurotrophism.
December 5, 2012|
Two species of songbirds pack their nests with scavenged cigarette butts that repel irksome parasites.
December 4, 2012|
Elderly people are worse at spotting untrustworthy faces, possibly due to decreased activity in the brain region associated with such perceptions.
December 1, 2012|
Meet some of the people featured in the December 2012 issue of The Scientist.
December 1, 2012|
The poxvirus stockpiles genes when it needs to adapt.
November 30, 2012|
Laurent Descarries, well known for his research on the brain's axon terminals, has passed away at the age of 73.
November 28, 2012|
In the largest microbial eukaryote genetic sequencing effort ever attempted, researchers are investigating the transcriptomes of 700 marine algae species.
November 28, 2012|
Rodents experience placebo-induced pain relief, providing a new model with which to investigate the phenomenon.


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This post was brought to you by Shout Factory who provided a media sample to facilitate my review. It was 1986. I was a Long Island girl with important things on my mind, like Duran Duran, purple eyeshadow and Stiff Stuff hairspray. Looming overheard? A move to Central Pennsylvania right before the start of my freshman… [read more]
The year was 1986… #PeeweesPlayhouse
The Jungle Book Diamond Edition: Now available! (and Printable Activities)
Cabin fever STILL in effect? Fear not – Today is the day that THE JUNGLE BOOK DIAMOND EDITION comes out on Blu-ray™, DVD and Digital. Once you grab a copy, keep the Disney fun going with printable activities, like a "Bare Necessities" quiz, character fun facts, "spot the difference" and more! Just click the image below to get them… [read more]
Stocking Stuffer Idea: The Littlest Pet Shop: Lights, Camera, Fashion (Now on DVD!)
I don't know what it is, but the little boys in my family all seem to looooove The Littlest Pet Shop! Lucky for Santa, there's a brand new DVD to bring this year, so our little guys are getting it (and I'm thinking their sisters won't be complaining. Could this be one DVD all… [read more]
Copy Kids Really DO Eat Fruits and Vegetables! DVD Giveaway {Ends 2/10}
Since the day I got that first pink line, my goal was to make sure my child was familiar with lots of different fruits and vegetables. I loved them as a kid, and I had high hopes that he would, too. And he always has. So, imagine my surprise the other night when he looked… [read more]


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(Updates with overseas alliances in the third paragraph.)
July 5 (Bloomberg) -- Nippon Life Insurance Co., Japan's biggest life insurer, will invest 500 million euros ($724 million) in a unit of Allianz SE as it expands overseas.
Nippon Life will buy bonds issued by the Allianz's unit that have rights to be converted into equities of the Munich- based parent company, Nippon Life said in a statement posted on its website today. The transaction, aimed at strengthening the two companies' capital ties, will be completed on July 7, the Japanese company said.
Nippon Life has been seeking alliances with overseas rivals to make up for slowing premium growth in the home market, where the average age is increasing at the world's fastest rate. The company, a mutual society owned by policyholders, agreed in March to buy a 26 percent stake in India's Reliance Life Insurance Co., marking the largest foreign direct investment in the South Asian nation's insurance industry.
Nippon Life, with head offices in Tokyo and Osaka, invested $250 million in Northwestern Mutual Life Insurance Co. last year and $500 million in Prudential Financial Inc. in 2009. Both companies are based in the U.S.
The investment in Northwestern Mutual was part of the two insurers' efforts to expand an alliance formed in July 2008 when Nippon Life bought a 5 percent stake in Russell Investments, a Northwestern subsidiary.
--Editors: Russell Ward, James Gunsalus
To contact the reporters on this story: Komaki Ito in Tokyo at email@example.com; Tomoko Yamazaki in Tokyo at firstname.lastname@example.com
To contact the editor responsible for this story: Andreea Papuc at email@example.com


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I don't even know where to begin asking for help. We have purchased four 40ft shipping containers which we are planning to transform into a home. We have the containers, and we will soon have the welder here to weld them together. We thought it would be a simple matter of welding them together, cutting out interior walls, and finishing it out just like a standard house. Well, standard with modifications dealing with working with steel walls.
The welder's first concern was how we planned to support the frames after removing a lot of the interior walls. These units are set side by side, and the plan was to have as open a living space as possible with the resulting 40 x 32 ft shell. The only interior walls we want will be for the bedrooms and bathrooms. Our plan leaves an open space for kitchen/living/dining room of 32 x 26 ft.
So, my first question is how DO we cut out a majority of the steel walls and keep it structurally strong? Pillars every 20 feet? Extra top rails welded? Can leaving say 6 inches of the walls in place serve as beams to keep the shells from bucking or caving in?
My questions go on and on, but first I have to figure out how to cut out the walls and still maintain the structural integrity.
Any advice, websites, book suggestions are greatly appreciated!!
- Forum >
- Miscellaneous >
- Residential project using steel containers
Thanks, yes I've watched the clips as well as several that are posted on youtube. But I haven't seen one that addresses my specific question, maybe I just missed it?
Used Shingles to Pave Driveway
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Notice Board
Internship Reflection Series
Dates: September 15, 22, 29, October 6, 5 to 6 p.m.
Location: CCC, Wright Hall 013 (lower level)
The CCC and the Wurtele Center for Work and Life present the Fall 2014 Community Engagement Internship Reflection Series for any students who have recently completed internships in the U.S. or abroad. During the sessions, you will engage in reflection and conversation about meaningful learning moments and receive guidance in writing a short reflective essay (500-700 words). Students have made use of this kind of writing in graduate school applications, job interviews, and newsletters--and you will likely find additional uses for it. Now that you've unpacked your bags, unpack your experience and discover what you learned!
Registration REQUIRED: Click here to register.
"Be A Better Tutor" Training
Dates: September 15, 23, October 6, 21, 27, November 4
Location: Dewey Common Room
The CCC is offering the Fall 2014 "Be A Better Tutor" Program, to enhance the skills of Smith students working (or planning to work) as tutors/mentors in area schools and after-school programs. Local educators will offer six lunchtime workshops that will focus on tutoring strategies and techniques. Students are welcome to attend one or more of the workshops; lunch will be provided.
Registration REQUIRED: Click here to register.
New: Community Engagement Field Guide!
Look for it and connect to information about volunteer placement opportunities, reflection sessions, leadership trainings, and upcoming events.
Community Engagement and Social Change (CESC) Concentration
The CCC invites students to find out
about the concentration in "Community Engagement and Social Change." This
concentration will help you link what you are learning in the classroom to the
needs and interests of communities locally and around the globe, and make a
difference with your Smith education. Please inquire early!
For information: Refer to smith.edu/ccc/concentration


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This Thomas Frankland ,born 30 April 1840 in Asenby was my husband Don's Gt.Grandfather and son of another Thomas Frankland, a wheelwright in 1841 and later listed as a Timber Merchant on his son's marriage entry in Helmsley in 1863 to Frances Hannah Strickland of Bransdale. There are Franklands who lived on the North Moors but though I have searched and noted all Franklands in the Kirkbymoorside, Rosedale and Thirkleby parish registers and in the I.G.I., I have not found that line of Franklands there.
Gt.Gt G.father Thomas Frankland was married to Ann ? about 1838 perhaps and both were born from about 1811-1816 .
We would like to find out the following :Ann's surname, place of marriage and date and place of each one's birth. They had two other children that we know of; Jane Frankland born about 1839 (aged 2 years in the 1841 Census) and Anne born in 1843 also in Asenby. This village was in the Topcliffe Parish . If anyone can help with these leads and/or with descendants of Jane and Anne,we would be immensely grateful. I may be able to help with identifying other Franklands in the areas which we have researched.
Don and Anne Frankland .
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Launcher Update
Get the Desktop App for Battle.net Now
- All your games in 1 place
- Log in once
- Automatic game updates
There should be a pop up quest for those that hit 90, and have no clue where to go.
In the mail with the reward quest with instructions to fly.
(I'm sorry, I saw the shiny firework, closed the mailbox and shot it off so fast...so maybe a separate letter would suffice.)
Long story short...
I hit 90. Pandaria is confusing. I got lost, repeatedly, just trying to get to Dread Wastes...
Luckily, I have a nice guild member with a mount I could ride on. He dragged me to the correct location to buy Pandarian Flight.
However, nothing is stopping me from ignoring the quest that was supposed to take me there to begin with. Then again, nothing is helping me.
Threats of violence. We take these seriously and will alert the proper authorities.
Posts containing personal information about other players. This includes physical addresses, e-mail addresses, phone numbers, and inappropriate photos and/or videos.
Harassing or discriminatory language. This will not be tolerated.


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Hi I am working on a project and need to get the type of logon for each user on the domain. By type I mean if the user connected to our domain via their phone to check e-mail, or if they are on a laptop and connected through a VPN, or just the plain desktop authentication.
I need this because we have a policy here that a person who has not logged on in over 10 days must be disabled. I am not seeing anything obvious on the MSDN site, nor anything relevant in any posts.
any help would be greatly appreciated. I am looking to do this in C#, as I already have a few other class libraries in C# doing some other AD functions and want to just add this into that solution.


text:
A fire caused heavy damage to an apartment building in Greenfield Wednesday afternoon, but everyone who was inside the building is believed to have escaped without injury, a fire department official said.
One firefighter was treated for heat-related stress during the fire at the 24-unit building near S. 88th St. and W. Layton Ave., Greenfield Fire Department Interim Chief Jon Cohn said.
Eight of the units sustained heavy damage and at least four sustained secondary smoke and water damage, Cohn said.
The fire appeared to have started in the rear of the structure, however the cause of the blaze was under investigation, Cohn said.
Fifteen residents were displaced by the fire and the Red Cross was called to the scene to assist them, Cohn said.
No further information on the fire was available late Wednesday afternoon.
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- Hales Corners Eagle Scout's signs point the way at Whitnall Park


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Find records of Wanda Andriuskevic
Find birth, death, marriage, military, and other records of Wanda Andriuskevic at Ancestry.com.Search for records
Biography & Family History Add details
Wanda Andriuskevic (1908 - 1992) was born on January 10, 1908. She was born into the Andriuskevic family.
There are no known spouses or children on file for Wanda. Wanda died in February 1992 at 84 years old.
Wanda Andriuskevic's last known residence is at Chicago, Cook County, Illinois.
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The 2014 Arizona Musicfest continues through March 1. Artistic Director, Robert Moody, dropped by K-BACH to chat about some of the highlights with K-BACH's Sterling Beeaff.
Arts and Culture
K-BACH's Theater Critic, Chris Curcio reviews the Phoenix Theatre production of the stage version of "Mary Poppins."
A new manifesto on the creative process from a master of the impossible-- "Creativity: The Perfect Crime" by Philippe Petit, the man who walked a high wire between the Twin Towers of the World Trade Center.
Prokofiev's Grandson imports Classical Club series to NYC; Met asks Singers to cut fees; Fleming retiring in 3 yrs?


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Thursday, Nov. 27 and Friday, Nov. 28 for Thanksgiving.
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Was here for New Year 2012. This is nice hotel, nice workers and the most important rooms are very clean. Hotel is near Prater and near u ban station so it is very easy to reachcity center and all other museums and places what you should see when you come to Vienna.
- Reservation Options:
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- Also Known As:
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A train depot built in 1916 in Novato, Calif., will soon don a new shingle roof. The Sonoma Marin Area Rail Transit Authority contributed about $6,000 to reroof the historic building, which has for 15 years prevented leaks with plastic tarps.
View Full Article in:Contra Costa Times (Walnut Creek, Calif.)
Published in Brief:


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Maybelline • Lash Stiletto - Waterproof • Mascara
(240 reviews)
|Average Rating.
|Would buy this product again.
*TOP REVIEWER* joy2uandme on 10/25/2014 4:57:00 PM more reviews by joy2uandme
Age: 56 & Over Skin: Sensitive, Fair-Medium, Neutral Hair: Brown, Straight, Fine Eyes: Hazel
Very similar to Lancome Definicil in performance for duh cheap. I got volume and length and it wore well. Nice glossy black color. Was looking for something that performed for cheaper and this exceeded my expectations. Kicking myself for buying expensive mascara.
Was this review helpful to you? Yes No
choanna on 8/28/2014 8:07:00 PM more reviews by choanna
Age: 19-24 Skin: Acne-prone, Fair-Medium, Neutral Hair: Brown, Straight, Fine Eyes: Brown
This is a great, simple mascara for everyday.
Mascaras smudge and/or flake on me without fail. ALL OF THEM.
But Lash Stiletto for some reason doesn't! It has very minimal to no smudging which is incredible for me.
This mascara is nice and black. It separated my think, long eyelashes well, and it gives light to medium volume- which is exactly what I like.
Mascara, just like makeup in general, is completely personal preference.
But if you want clean, separated, lengthened, very shiny black, and slightly voluminous eyelashes, you might want to try Maybelline Lash Stiletto! :)
(btw the packaging is beautiful; I really like it!)
Now it's October 10th so about a month and a half and I want to update everyone on this mascara!
So far... I have to say since it started off as a pretty dry mascara (which I love) it's been a little bit too dry... but that's too be expected with drier mascaras.
It's still usable and it still does not smudge for flake off, so I have to give it credit for staying true to its claims even over time.
I still love this mascara, and I'll keep using it as long as I can brush through my eyelashes after application with a clean wand to even everything out!
Just please keep in mind that because this mascara is already dry in the beginning, it will seen to dry faster than wet mascaras
When this becomes unusable I'll go out and get a new one!
I'll update this review when it finally goes crumbly and unusable
1 out of 1 people found this review helpful. Was this review helpful to you? Yes No
prettyBOSSet on 7/1/2014 3:11:00 AM more reviews by prettyBOSSet
Age: 19-24 Skin: Normal, Medium Hair: Brown Eyes: Brown
I love this mascara. I naturally have thick, long lashes and I needed a product to help tame them and make them more defined. This product works wonders.
Was this review helpful to you? Yes No
makeuprubies on 6/7/2014 2:12:00 AM more reviews by makeuprubies
Age: 19-24 Skin: Combination, Fair, Cool Hair: Brown, Wavy, Fine Eyes: Green
I really like this mascara. I bought it for NZD $9 at a makeup sale which is a decent price for mascara. I was sceptical as the brush was smaller than ones id normally use (Big and fat for volume) but the brush actually gives a nice fanned look on my lashes, and seperates really well. Holds my curl and gives awesome length. Would repurchase :)
1 out of 1 people found this review helpful. Was this review helpful to you? Yes No
Laurynyumi on 5/23/2014 6:14:00 PM more reviews by Laurynyumi
Age: 30-35 Skin: Combination, Olive Hair: Black Eyes: Brown
Since you have to replace mascara so often I don't really understand why I need to buy a super expensive mascara everytime. I really like this mascara I have Asian eyes and not many lashes, the few I have this mascara really lengthens them. This is my fav mascara right now.
Was this review helpful to you? Yes No
misscynthia06 on 5/16/2014 4:49:00 AM more reviews by misscynthia06
Age: 25-29 Skin: Combination, Tan Hair: Brunette Eyes: Brown
This is my HG bottom lash mascara.
The brush is long and skinny and the formula not too wet, so perfect to use on my tiny bottom lashes.
This stuff is waterproof and does not budge on my watery eyes, yet is easy to remove with makeup remover. This is very exciting as some waterproof mascara is so so hard to remove.
I really don't use this product on my top lashes, the effect is fine, long, defined and separated lashes, I just really prefer a more voluminous look.
1 out of 1 people found this review helpful. Was this review helpful to you? Yes No
lilyde123321 on 5/11/2014 5:32:00 PM more reviews by lilyde123321
Age: 25-29 Skin: Oily, Olive Hair: Other Eyes: Other
This product is a miracle in a bottle! It noticeably lengthens and defines lashes, does not clump, does not smudge, and it is easy to remove. Best of all, you can find it at any drug store and it is cheap! Winning!
1 out of 1 people found this review helpful. Was this review helpful to you? Yes No
PatentlyPink on 3/4/2014 10:49:00 PM more reviews by PatentlyPink
Age: 25-29 Skin: Combination, Fair-Medium, Cool Hair: Brown, Straight, Coarse Eyes: Brown
Great drugstore mascara, my favorite at the moment. Easy to get volume, length and a great curl. Stayed well during a session of sweaty yoga. Cheap, cute name, nice packaging. Another maybelline winner!
Was this review helpful to you? Yes No
BP8099 on 2/23/2014 1:57:00 PM more reviews by BP8099
Age: 25-29 Skin: Combination, Fair Hair: Brown Eyes: Green
I love this mascara. it doesn't smudge or flake.
Was this review helpful to you? Yes No
cjhalfthai on 2/18/2014 2:28:00 AM more reviews by cjhalfthai
Age: 25-29 Skin: Normal, Fair-Medium Hair: Black, Wavy, Coarse Eyes: Brown
This is my favorite cheap drugstore mascara. I always purchase one tube after the other because I like the effect it gives on my lashes. It lengthens my lashes, it has fibers to extend my lashes. Unfortunately, these fibers fall out and leave my fave with fine fibers on my cheeks. But I really love how it makes my eyelashes look naturally long. It doesn't give you the spider-leg effect. ;)
Top Rated Mascara
- 132 reviews
Longcils Boncza Cake Mascara in Black39 reviews
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Lumene Blueberry curl mascara53 reviews


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The Tablas Creek Vineyard 2011 Full Circle Pinot Noir is Tablas Creek's second bottling of this renowned Burgundy grape from the small vineyard outside Robert Haas's family home in Templeton. We named the wine Full Circle because it reflects his career: from a start introducing America to the greatness of Burgundy, through decades focusing on grapes from the Rhone, he's now growing Pinot at home.
After importing our Châteauneuf du Pape clones, we brought in selections of a few other high quality (non-Rhone) clones as part of an effort to expand our nursery business, including Pinot Noir. Although we eventually decided that our nursery should remain focused solely on the Rhone grape varieties we grow, we planted two rows of Pinot Noir near our nursery to produce enough vine material for the 2.5 acre vineyard around founder Robert Haas's house in Templeton. The Templeton Gap has been long recognized for its marine influence and resulting microclimate that is the coolest in the Paso Robles AVA, and the Haas Vineyard is in one of the coolest pockets of Templeton, near Santa Rita Creek. This vineyard is farmed organically by the Tablas Creek Vineyard team.
The 2011 vintage was our second consecutive winter with healthy rainfall, but yields were dramatically reduced by two nights of frost on April 8th and 9th. Despite the low crop loads, ripening was slow due to a very cool summer, and harvest roughly three weeks later than normal, beginning in mid-September and not concluding until mid-November. Warm, sunny weather during harvest allowed the later-ripening varieties to reach full maturity. The long hangtime and low yields combined to produce fruit with notable richness balanced by higher than usual acidity. The Haas Vineyard Pinot Noir was harvested in two picks in late September and early October.
The grapes were fermented in one-ton microfermenters using native yeasts. After pressing, the wine was moved into year-old Marcel Cadet 60-gallon barrels, for a hint of oak. The wine stayed on its lees, stirred occasionally, for a year and a half before being blended and bottled in April 2013.
The 2011 Full Circle shows shows a classic, vibrant Pinot Noir aromas of sweet spices, black tea, plum and earth. The mouth leads with loamy minerality and follows with purple fruit, good acids and granular tannins. Dark cherry, chalky minerality and spice linger on the finish. Drink now or over the next decade.
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On December 6th, all VINsiders are invited to join Tablas Creek's owners and winemakers for our 10th annual En Primeur Tasting and Futures Offering. Taste the 2013 Esprit de Tablas and 2013 Panoplie, discuss the vintage, and enjoy a seasonal dish created to pair with the wines. The $30 cost to attend the event is refundable on any en primeur purchase. Register for one of two sessions (10:30am or 1:30pm) More information »
This holiday season, choose from 5 gift packages, all with shipping included in a gift box. We even have an epicurean pack including our organic estate olive oil that we can ship to all 50 states! Details & more gift ideas »
We were thrilled to receive such good reviews from this notoriously tough grader, including the 2012 Esprit de Tablas Blanc (92 points; "impeccably refined"), 2011 Panoplie (94 points; "pure elegance"), 2012 Patelin de Tablas (90 points; "a gorgeous wine and a fabulous value"), and the 2012 Esprit de Tablas (93-95 points; "a fascinating Esprit to follow over the coming years"). Read the complete review » More recent press »
November, 21 2014
Over recent weeks, I've received several questions from people wondering how to best preserve a partial bottle of wine for future consumption. Typically, they're wondering if they should invest in a system that replaces the air in the bottle with an inert gas, or in a vacuum system that removes the air from the bottle entirely... Read More »


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Originally published October 15, 2013 at 5:48 AM | Page modified October 15, 2013 at 6:54 AM
European Space Agency aims to 'print' spacecraft parts using new technology
The European Space Agency says it has developed a technology that allows metal parts for spacecraft and nuclear reactors to be "printed" as a single piece.
The European Space Agency says it has developed a technology that allows metal parts for spacecraft and nuclear reactors to be "printed" as a single piece.
In recent years three-dimensional printing has become commonplace in manufacturing. But so far this method has largely been limited to making plastic objects.
ESA says its metal-printing technology can be used to create more complex shapes than with traditional techniques and leaves almost no waste material.
The Paris-based space agency presented several sample objects capable of withstanding temperatures of up to 1,000 degrees Celsius (1,832 Fahrenheit) at an event in London on Tuesday.
It aims to make the parts even more durable in the coming years.
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