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Infectious diseases represent the greatest threats to endangered species, and transmission from humans to wildlife under increased anthropogenic pressure has been always stated as a major risk of habituation.
To evaluate the impact of close contact with humans on the occurrence of potentially zoonotic protists in great apes, one hundred mountain gorillas (Gorilla beringei beringei) from seven groups habituated either for tourism or for research in Volcanoes National Park, Rwanda were screened for the presence of microsporidia, Cryptosporidium spp. and Giardia spp. using molecular diagnostics.
The most frequently detected parasites were Enterocytozoon bieneusi found in 18 samples (including genotype EbpA, D, C, gorilla 2 and five novel genotypes gorilla 4–8) and Encephalitozoon cuniculi with genotype II being more prevalent (10 cases) compared to genotype I (1 case). Cryptosporidium muris (2 cases) and C. meleagridis (2 cases) were documented in great apes for the first time. Cryptosporidium sp. infections were identified only in research groups and occurrence of E. cuniculi in research groups was significantly higher in comparison to tourist groups. No difference in prevalence of E. bieneusi was observed between research and tourist groups.
Although our data showed the presence and diversity of important opportunistic protists in Volcanoes gorillas, the source and the routes of the circulation remain unknown. Repeated individual sampling, broad sampling of other hosts sharing the habitat with gorillas and quantification of studied protists would be necessary to acquire more complex data.
Citation: Sak B, Petrželková KJ, Květoňová D, Mynářová A, Pomajbíková K, Modrý D, et al. (2014) Diversity of Microsporidia, Cryptosporidium and Giardia in Mountain Gorillas (Gorilla beringei beringei) in Volcanoes National Park, Rwanda. PLoS ONE 9(11): e109751. https://doi.org/10.1371/journal.pone.0109751
Editor: Yung-Fu Chang, Cornell University, United States of America
Received: July 1, 2014; Accepted: September 11, 2014; Published: November 11, 2014
Copyright: © 2014 Sak et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Data Availability: The authors confirm that all data underlying the findings are fully available without restriction. All relevant data are within the paper.
Funding: This work was supported by the project "CEITEC" - Central European Institute of Technology (CZ.1.05/1.1.00/02.0068) from the European Regional Development Fund, co-financed from the European Social Fund and the state budget of the Czech Republic (project OPVK CZ.1.07/2.3.00/20.0300), by a grant from the Grant Agency of the Czech Republic (206/09/0927), and by institutional support of Institute of Vertebrate Biology Academy of Sciences of the Czech Republic (RVO:68081766). K.J.P. was also supported by the Praemium Academiae award to Julius Lukes. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exists. The employment of authors does not alter adherence to PLOS ONE policies on sharing data and materials.
Microsporidia, Cryptosporidium spp. and Giardia spp. are unicellular parasites spread by the fecal-oral route by environmentally resistant stages and they can infect humans, livestock and wildlife animals including non-human primates [1]–[5]. The symptoms in immunocompetent hosts are usually mild and self-limiting. However, human cryptosporidiosis, microsporidiosis and giardiosis emerged as important opportunistic diseases when AIDS became pandemic [6], [7]. Thousands of HIV-infected patients with chronic diarrhea attributed to these organisms have been reported from all over the word [7], [8].
Recent studies surprisingly showed high prevalence of asymptomatic microsporidial infection in captive and habituated great apes [3], [5], while Cryptosporidium and Giardia spp. seems to be less frequent pathogen [1], [3], [9]. It has been suggested, that increased prevalence of these pathogens could be a result of frequent presence of humans and livestock in the ranges of primates [4], [9]–[11], rather than resulting from close contact with humans [12]. Moreover, as opportunistic pathogens they are not strictly host specific and hence able to cross the species transmission barrier [13]–[16]. However, only genotyping and subtyping of particular isolates can provide the essential information about epidemiology and zoonotic potential of these protists in primates [11], [17]–[19].
The mountain gorilla (Gorilla beringei beringei) is classified as an endangered species with only two isolated populations remaining in Bwindi Impenetrable National Park, Uganda (330 km2) and the Virunga Massif (450 km2) at the borders of Rwanda, Uganda, and the Democratic Republic of Congo [20], [21]. Observing habituated mountain gorillas has proven particularly popular since it began in the 1950s, and is one of the world's best-known wildlife experiences [22]. During the habituation process animals become accustomed to human presence and are thought eventually to accept a human observer as a neutral element in their environment [23]. However, the consequences of increasing human-gorilla contacts in habituated groups can have negative effects on animals [24]–[29]. An increased anthropogenic impact on primate populations may result in general changes in communities of their parasites, and also in a direct exchange of parasites between humans and primates [30], [31].
Since only limited work has been done to explore the molecular diversity of gastrointestinal parasites in mountain gorillas [1], [2], [32]–[34] we conducted a comprehensive molecular screening for Encephalitozoon spp., Enterocytozoon bieneusi, Cryptosporidium spp. and Giardia spp. in several groups of habituated mountain gorillas (Gorilla beringei beringei) in Volcanoes National Park, Rwanda.
Materials and Methods
Ethics Statement
The research complied with the legal requirements of Rwanda and adhered to the research protocol submitted to conservation authorities. The permission to collect the fecal samples was obtained from Rwanda's Office of Tourism and National Parks. Since the collection of fecal samples from gorillas was noninvasive and did not cause any observable distress to the animals, no animal ethic committee was consulted regarding our study. The sampling was performed during routine health checks by Mountain Gorilla Veterinary Project and Dian Fossey Gorilla Fund employees. No interaction with animals was conducted for this study.
Study site
The study was conducted in Volcanoes National Park in North western Rwanda, a montane rain forest that ranges in altitude from 2300 to 4500 m and supports distinct vegetation communities at different elevations [35], [36]. The area is surrounded by some of the highest rural human population densities in the world, up to 820 people per km2 [37]. High densities of humans can have negative impact on conservation of wildlife in the Park [38]. Virunga subpopulation of mountain gorillas have suffered numerous threats such as habitat destruction for firewood and agriculture, illegal cattle grazing, illegal logging and illegal poaching [39]. Although G. beringei beringei was affected by war and instability, currently the park has undergone through significant regeneration, with investment into tourism, improvement of social infrastructure and safety [40], [41].
Studied gorilla groups and sample collection
In September 2007, we collected 100 individual fecal samples from night nests of seven gorilla groups that were habituated and used either for tourism (5) or for research (2) (Table 1). Usually four trackers and up to four researchers visit the research groups for approximately four hours every day. The tourist groups are visited by four local trackers, two local guides and up to eight tourists for an hour daily at high booking seasons. People following both tourist and research groups are accompanied by military men and porters who stay approximately 100 yards away from gorillas for the safety reasons. In 2007, Volcanoes NP Rwanda has reported 18,000 tourists visiting the Park [42].
To prevent the repeated sampling of same individuals, which could distort the parasite prevalence estimation, each group was sampled only once. All fecal samples were immediately preserved in 96% ethanol and transported to the Institute of Parasitology, Biology Centre of Academy of Sciences of the Czech Republic.
DNA extraction, PCR amplification, sequencing and genotyping
The suspension of each fecal sample in alcohol was evaporated overnight at 60°C. A total of 200 mg of fecal samples were homogenized by bead disruption using 0.5 mm glass beads (Biospec Products, Inc., Bartlesville, OK, USA) in a FastPrep-24 Instrument (MP Biomedicals, Santa Ana, CA, USA) at a speed of 5 m/s for 1 min followed by isolation/purification using the QIAamp DNA Stool Mini Kit in accordance with the manufacturer's instructions (Qiagen, Hilden, Germany). Purified DNA was stored at −20°C prior to use in PCR. All DNA samples obtained for the study were analyzed by polymerase chain reaction (PCR) using sets of specific primers. A nested PCR approach was used to amplify a region of the internal transcribed spacer (ITS) of Enterocytozoon bieneusi (∼390 bp) [43], the small ribosomal subunit rRNA (SSU) gene of Cryptosporidium spp. (∼830 bp) [44], and the triosephosphate isomerase (TPI) gene of Giardia intestinalis (also called Giardia lamblia, Giardia duodenalis) (∼500 bp) [45]. The following primers sets were used to amplify Encephalitozoon spp.: the int580f and int580r primer set for primary PCR analysis [46] and the MSP3 and MSP4 primer set for secondary PCR (∼320 bp) [47]. Secondary PCR products were run on a 2% agarose gel containing 0.2 µg/ml ethidium bromide in 1× TAE buffer at 75 volts for approximately 1 hour. Bands of the predicted size were visualized using a UV light source, cut from the gel, and then extracted using QIAquick Gel Extraction Kit (Qiagen, Hilden, Germany). Gel purified secondary products were sequenced in both directions with an ABI 3130 genetic analyzer (Applied Biosystems, Foster City, CA, USA) using the secondary PCR primers and the BigDye1 Terminator V3.1 cycle sequencing kit (Applied Biosystems, Foster City, CA, USA) in 20 µl reactions.
Positive and negative controls were included in each analysis. DNA from E. intestinalis spores grown in vitro in the Laboratory of Veterinary and Medical Protistology at the Institute of Parasitology of ASCR, from E. bieneusi spores of genotype S6 originally isolated from a eastern house mice, from Cryptosporidium serpentis originated from corn snake, and from Giardia intestinalis assemblage E originated from the domestic goat were used as positive control for appropriate PCR. All samples were analyzed in duplicate. In case of positive detection, the sample was newly re-isolated and previous finding was independently verified.
Phylogenetic analyses
The nucleotide sequences of each gene obtained in this study were edited using the software ChromasPro 1.5 (Technelysium, Pty, Ltd., Brisbane, Australia) and were aligned with each other and with reference sequences from GenBank using ClustalX 2.0.6. Alignment adjustments were made manually to remove artificial gaps using BioEdit. Phylogenetic analyses were performed using the software MEGA5 [48]. Neighbour joining (NJ) trees were constructed. All ambiguous positions were removed for each sequence pair. The reliability of branches in trees was assessed using the bootstrap analysis with 1000 pseudo-replicates, with values above 50% reported. Phylograms were drawn using the MEGA5 and were manually adjusted using CorelDrawX5 (Ottawa, Canada). ITS and SSU sequences have been deposited in GenBank under the accession numbers KJ469967–KJ469979, respectively.
Statistical analyses
To analyze the differences in microsporidia (Enterocytozoon spp, Encephalitozoon spp.) infections between groups habituated for tourism and research we fitted the generalized linear mixed model (GLMM) with binomial distributions. Samples were classified according to the "group" (Pablo, Shinda, Amahoro, Umubano, Sabinyo, Susa, Kwitonda) and "habituation" (research, tourist). The random factor "group" was nested into the fixed factor "habituation". Statistical analyses were conducted in R 2.13.1. [49].
Out of the total of 100 examined gorilla samples 33 were positive for tested parasites. The most frequently detected parasites were E. bieneusi in 18 and Encephalitozoon cuniculi in 11 samples. Cryptosporidium spp. was identified in 4 samples and Giardia spp. in none of them (Table 1).
The alignment of the obtained microsporidial ITS sequences with reference sequences showed 100% homology with GenBank-listed species and their genotypes as follows: 10 E. cuniculi genotype II (GQ422153) and 1 E. cuniculi genotype I (AF338410) (Table 1). A phylogenetic analysis of all ITS sequences performed on a multiple alignment that included representatives of E. bieneusi genotypes accessible in GenBank revealed the presence of sequences matching with previously described human pathogenic E. bieneusi genotype D (JF927954) in three cases, seven EpbA (AF135833) and two E. bieneusi genotype C (AF101199). Moreover, one genotype previously described from western lowland gorillas, gorilla 2 (JQ837794) was identified in one animal. Furthermore, five sequences of our isolates belonged to five novel genotypes, named gorilla 4–8. While two new genotypes gorilla 7 and gorilla 8 clustered closely to genotype EpbA and C, respectively, genotypes gorilla 4–6 formed a group with isolates from humans and pigs. The global topology of the tree is shown in Fig. 1.
The host is listed for each sample. Values on branches are percent bootstrapping using 1 000 replicates. The bootstrap proportions greater than 50% are shown at each branch. Nucleotide sequences generated from this study are underlined and are deposited in the GenBank under Accession Numbers KJ469967–KJ469979.
Phylogenetic analyses based on SSU sequences showed that Cryptosporidium originating from the Pablo group were 100% similar to C. meleagridis (AF112574) and those from Shinda group to C. muris (AB089284) (Table 1).
Single-species infection was detected in most animals with the exception of three individuals co-infected with C. meleagridis and E. cuniculi genotype II, C muris and E. bieneusi genotype D and E. cuniculi genotype II and E. bieneusi genotype gorilla 2, respectively (Table 1).
Cryptosporidium infections were found only in animals from research groups. We did not detect any differences in the occurrence of Enterocytozoon bienusi between research and tourist groups (GLMM, treatment contrasts given: z = –0.086 p = 0.932), but the Encephalitozoon cuniculi was more prevalent in research groups compared to tourist ones (GLMM, treatment contrasts given: z = –2.742, p = 0.006). However, the four of five novel genotypes of E. bieneusi were found in research groups (Table 1).
To our knowledge, this is the first study providing detailed molecular information about Cryptosporidium spp., Encephalitozoon cuniculi and Enterocytozoon bieneusi infections in wild mountain gorillas in the Virunga Volcanoes region.
While our results show that microsporidia, especially various E. bieneusi genotypes reached a prevalence of 29%, results of previous studies revealed that 11% of gorilla samples from Bwindi NP contained Cryptosporidium parvum compared to E. intestinalis in 3% and G. intestinalis in 2% of samples [1]–[3], [33]. On the contrary, E. cuniculi was detected as predominant species among western lowland gorillas in the Dzanga-Sangha Protected Areas in Central African Republic [12].
We did not detect any species of protists previously identified in mountain gorillas, namely Giardia intestinalis, Cryptosporidium parvum and Encephalitozoon intestinalis [1]–[3], [32]–[34]. Although most of microsporidia species detected in our study (with exception of the novel genotypes) were already detected in western lowland gorillas [12], C. muris and C. meleagridis were identified in gorillas for the first time.
We detected 9 different genotypes including E. bieneusi genotype EbpA, D, and C described from humans, non-human primates from zoo's and domestic animals including pigs and cattle [5], [50]–[54]. In previous study only four distinct genotypes of E. bieneusi, including genotype D and gorilla 1–3, have been described from western lowland gorillas with genotype D as predominant [12].
All five new E. bieneusi genotypes from this study (gorilla 4–8), together with genotype gorilla 2 reported in western lowland gorillas, were genetically related to genotype D and belonged to group 1, as identified by Thellier and Breton [55]. Genotypes of this group are known to have zoonotic potential and they are of public health significance [55]. However, the potential role of humans and domestic animals as a source of infection for mountain gorillas remains speculative, since no current data about microsporidia in hosts sharing habitat with gorillas were available.
We identified E. cuniculi genotypes I and II with genotype II as the most prevalent. The same genotypes of E. cuniculi have also been detected in both free-ranging western lowland gorillas and captive chimpanzees from several zoos and sanctuaries, though the E. cuniculi genotype I was the most prevalent one in previous studies [5], [12]. Since E. cuniculi genotypes are not host-specific and have been identified in a variety of animal hosts such as rodents, lagomorphs, carnivores, horses, human and nonhuman primates, and birds [12], [53], [56]–[60], detection of the exact origin of E. cuniculi infections in gorillas is difficult.
Surprisingly we did not detect any E. intestinalis in our sample set, although it is the only microsporidium species that has been previously identified in mountain gorillas and people who shared habitats with them in Uganda [2]. However, we cannot exclude that gorillas we studied do not have also concurrent E. intestinalis infection as diagnosis of mixed infections is limited by amplification of the predominant species over the others as it was observed in Cryptosporidium spp. mixed infections [61]. Additionally, our results are hampered by the one-shot sampling methodology, which could not provide precise estimation of prevalence due to intermittent shedding of infectious stages of parasites into the environment, especially in the case of Giardia sp. and microsporidia. A similar situation has been described in other hosts including humans [62]–[64].
We did not amplify any of Cryptosporidium species/genotypes previously detected in gorillas. While Graczyk et al. [33] identified zoonotic Cryptosporidium parvum genotype 2 in mountain gorillas, and Sak et al. [12] reported C. bovis in western lowland gorillas, we detected C. meleagridis and C. muris in two of examined gorillas. Typical host spectrum of C. meleagridis includes birds from orders Galliformes, Columbiformes, Psittaciformes and Passeriformes [65]–[67], while C. muris (in several genotypes) parasitizes rodents and artiodactyls [68].
No case of non-human primate infection by C. meleagridis has been reported so far and C. muris-like infection has been previously reported only in stomach of cynomolgus monkeys (Macaca fascicularis) [69]. However, both these cryptosporidia species are known to infect "non-typical" hosts [70], [71], being reported also from immunocompetent and immunodecificient humans. The source of cryptosporidial infection in mountain gorillas reported in this study is unknown, however, it is possible that under certain conditions this host specific cryptosporidia can be transmitted among species [70], [71], especially because both detected species has been previously reported to be human pathogenic in both immunocompetent and immunodecificient human patients. C. meleagridis is the third most common causative agents of human cryptosporidiosis in the world [72], whereas C. muris was detected only occasionally [73]–[77].
Our results imply that gorillas habituated and used for research were significantly more parasitized with E. cuniculi compared to gorillas habituated and used for tourism. Also Cryptosporidium sp. infections were found only in research groups. It might imply that research activities could pose greater risk than tourism, for example animals from research groups might be more stressed (and thus immunosuppressed) because perhaps people spend longer period with them (four hours per day) in comparison to tourist ones (one hour per day), but preliminary studies did not show any difference in fecal cortisol levels between. Noteworthy, Pablo and Shinda (as well as Susa, which is a tourist group) ranged nearby the southern edge of the Park, where more illegal activities occur [78]. This can also lead to increased stress and possibly increased risk of pathogen transmissions due to more frequent uncontrolled human presence in gorilla habitats. A closer look to our results revealed that these three groups are infected by E. cuniculi II while the others harbor genotype I; also the novel genotypes of E. bieneusi occurred only in Susa, Pablo and Shinda (Table 1). The overlapping habitat also raises the question of other animals (e.g. buffalo or elephants) or water sources, which can serve as reservoirs, but there are no apparent differences between the areas where these groups range and the rest of the Park (Cranfield, pers. obs.).
Our data showed the presence and diversity of important opportunistic protists in Volcanoes gorillas. However, the source, the routes of the circulation and the importance of these pathogens for health of gorillas remain in mist. Answering the persisting questions apparently requires bigger effort, involving: (i) broad sampling of other hosts sharing the habitat with gorillas, (ii) repeated sampling of individuals and (iii) quantification of studied protists and associating the results with actual health parameters of sampled individuals. Having all these data together is crucial for development of well-managed ecotourism and research activities with minimal impact on the animal health.
We would like to thank Antoine Mudakikwa and the management of the Rwanda Development Board for allowing this study to take place. The trackers of Karisoke Research Center and the Rwanda Development board aided with the collection of the samples. Gorilla Doctors veterinarians helped with the collection and processing of the samples. The authors thank Chrispher Whittier from Cummings School of Veterinary Medicine at Tufts University, for his helpful and constructive comments on the manuscript.
Author Contributions
Conceived and designed the experiments: KJP DM BS MC KP A. Mudakikwa. Performed the experiments: BS MK A. Mynarova DK. Analyzed the data: MK BS KJP. Contributed reagents/materials/analysis tools: MK BS KJP KP. Contributed to the writing of the manuscript: BS KJP MK MC DM.
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text:
Price Beat Guarantee*
Find a lower comparable hotel price online and we'll beat it by 10%. Find out more.
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Property Details
Property Location
When you stay at The Frutta Boutique Patong Beach in Patong, you'll be by the sea, within a 5-minute drive of Bangla Road and Jungceylon Shopping Center. This hotel is 1.6 mi (2.7 km) from Patong Beach and 4.4 mi (7.1 km) from Karon Beach.Rooms
Make yourself at home in one of the 60 air-conditioned rooms featuring refrigerators and LCD televisions. Complimentary wireless Internet access keeps you connected, and satellite programming is available for your entertainment. Private bathrooms with showers feature rainfall showerheads and hair dryers. Conveniences include phones, as well as safes and minibars.Amenities
Enjoy recreation amenities such as an outdoor pool or take in the view from a terrace. Additional features at this hotel include complimentary wireless Internet access, concierge services, and babysitting/childcare (surcharge). Guests can catch a ride to nearby destinations on the area shuttle (surcharge).Dining
Enjoy a meal at the restaurant or snacks in the hotel's coffee shop/café. Wrap up your day with a drink at the bar/lounge. Continental breakfasts are available daily from 8 AM to 10 AM for a fee.Business, Other Amenities
Featured amenities include express check-in, dry cleaning/laundry services, and a 24-hour front desk. Guests may use a roundtrip airport shuttle for a surcharge, and free self parking is available onsite.Attractions
Distances are displayed to the nearest 0.1 mile and kilometer.
Patong Beach - 1.5 km / 0.9 mi
Patong Boxing Stadium - 1.6 km / 1 mi
Royal Paradise Night Market - 1.6 km / 1 mi
Kalim Beach - 1.6 km / 1 mi
Bangla Boxing Stadium - 1.7 km / 1 mi
Bangla Road - 1.8 km / 1.1 mi
Banzaan Fresh Market - 1.9 km / 1.2 mi
Jungceylon Shopping Center - 2.1 km / 1.3 mi
Patong Go-Kart Speedway - 3.2 km / 2 mi
Tiger Kingdom Phuket - 3.2 km / 2 mi
Simon Cabaret - 3.6 km / 2.2 mi
Phuket Cable Ski and Phuket Wake Boarding - 5.7 km / 3.6 mi
Loch Palm Golf Club - 6.2 km / 3.9 mi
Freedom Beach - 6.4 km / 4 mi
Phuket Country Club - 6.5 km / 4 mi
The preferred airport for The Frutta Boutique Patong Beach is Phuket (HKT-Phuket Intl.) - 37.6 km / 23.3 mi
Property Policies
Extra-person charges may apply and vary depending on property policy.
Government-issued photo identification and a credit card, debit card, or cash deposit are required at check-in for incidental charges.
Special requests are subject to availability upon check-in and may incur additional charges. Special requests cannot be guaranteed.
- Please note that cultural norms and guest policies may differ by country and by property. The policies listed are provided by the property.
Additional fees of THB 300 apply to children ages 4-11 when sharing existing bedding with an adult. For more details, please contact the property using the information on the reservation confirmation received after booking.
Important Information
Please note, there is important information about this property to know before you arrive.
The following fees and deposits are charged by the property at time of service, check-in, or check-out.
- Fee for continental breakfast: THB 150 per person (approximately)
- Rollaway beds are available for an additional fee
The above list may not be comprehensive. Fees and deposits may not include tax and are subject to change.
- -In-room accessibility
- -Television in common areas
- -Bar/lounge
- -Breakfast available (surcharge)
- -Coffee shop or café
- -Safe-deposit box at front desk
- -24-hour front desk
- -Concierge services
- -Porter/bellhop
- -Smoke-free property
- -ATM/banking
- -Terrace
- -Grocery/convenience store
- -Free WiFi
- -Airport transportation
- -Outdoor pool
- -Laundry facilities
- -Dry cleaning/laundry service
- -Free self parking
- -Area shuttle (surcharge)
- -Tours/ticket assistance
- -Babysitting or childcare (surcharge)
- -Parking (limited spaces)
- -Restaurant
- -Beach towels
- -Elevator
- -Coffee/tea in common areas
- -Shopping center shuttle (surcharge)
- -Luggage storage
- -Airport transportation (surcharge)
- -Express check-in
- -Wheelchair accessible path of travel
- -Wheelchair accessible parking

subset: commoncrawl


text:
Elements of a Suitable Wellness Center
You will learn that experts will time and again play a major role in our quest to lead a healthier, stress-free and inspired life. It is evident that a wellness center will time and again be worth considering in this pursuit. They usually have all the right professionals to ensure that you get the services and results that you need. Nevertheless, selecting such wellness facilities can be a great challenge to come people. We have certain factors that you are expected to consider in this respect. Such will time and again entail the following.
Seek to ensure that you clearly spell out your goals in this pursuit. You will note that an ideal wellness center will often deal with so many health issues. This means that you need to be certain of what you want so as to pick the best. There are certain wellness centers that are more prominent in particular services. Always ensure that you opt for a wellness center that is ready to offer you what you are in need of. It is valuable for you to opt for a center that has the capacity to offer a variety of services under one roof. This is due to the kind of convenience that it comes with. Make sure that you are aware of the time that you will have to spend at the center. You need to find it quite comfortable. Ensure that you choose a duration that will assure you of wholesome revitalizing. It is through this that you will be assured of better returns on your investment.
Seek to ensure that you are aware of the staff that the center has. Make sure that you pick a center that has a wider range of specialists. You will be assured of better results from persons that are adequately skilled. This staff must be known for enriched professionalism as well as communication skills. This will often mirror the kind of comfort to expect in the long run. It is also important for you to consider the location of the center. You will appreciate a location that you can reach with much ease. The atmosphere within which it is set is expected to be quite peaceful. Save yourself from locations that are likely to compromise your comfort while undergoing the wellness program.
Cost often matters a lot in various issues. As such, getting the best price will be ideal for you. Take time to compare the rates offered by various providers to ensure you get an affordable deal.
The Beginner's Guide to Resources


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No arrests have been made.
She was the 16th trans woman to be killed in the U.S. this year.
The attack follows a recent spike in homophobic incidents.
The school is accused of ignoring pleas for help.
The incident highlights need for robust hate crime protections
A new report has seen a steady increase under Trump.
Police are investigating, though they aren't calling the attack as a hate crime.


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Hasanabadi, T., Habibi, D., Khalaj, H. (2016). Influence of Zeolite and Biological Fertilizer under Different Irrigation Regime on Quantitative and Qualitative Traits of Sugar beet (Beta Vulgaris L.).
Journal of Crop Nutrition Science
, 2(Issue 1-2), 20-31.
Tahereh Hasanabadi; Davood Habibi; Hamideh Khalaj. "Influence of Zeolite and Biological Fertilizer under Different Irrigation Regime on Quantitative and Qualitative Traits of Sugar beet (Beta Vulgaris L.)".
Journal of Crop Nutrition Science
, 2, Issue 1-2, 2016, 20-31.
Hasanabadi, T., Habibi, D., Khalaj, H. (2016). 'Influence of Zeolite and Biological Fertilizer under Different Irrigation Regime on Quantitative and Qualitative Traits of Sugar beet (Beta Vulgaris L.)',
Journal of Crop Nutrition Science
, 2(Issue 1-2), pp. 20-31.
Hasanabadi, T., Habibi, D., Khalaj, H. Influence of Zeolite and Biological Fertilizer under Different Irrigation Regime on Quantitative and Qualitative Traits of Sugar beet (Beta Vulgaris L.).
Journal of Crop Nutrition Science
, 2016; 2(Issue 1-2): 20-31.
Influence of Zeolite and Biological Fertilizer under Different Irrigation Regime on Quantitative and Qualitative Traits of Sugar beet (Beta Vulgaris L.)
Article 3
Volume 2, Issue 1-2
, Spring 2016
, Page
Tahereh Hasanabadi
Davood Habibi
Hamideh Khalaj
Young Researchers and Elites Club, Tehran North Branch, Islamic Azad University, Tehran, Iran.
Department of Agronomy and Plant Breeding, Karaj Branch, Islamic Azad University, Karaj, Iran.
Department of Agriculture, Payame-Noor University, P.O. Box: 19395-3697, Tehran, Iran.
In order to study the effect of zeolite and biologic fertilizers application under different irrigation regime on yield and quantitative traits in sugar beet, a research project was conducted according split-split plot experiment based on randomized complete block design with four replicates. The main factor included irrigation regime at two levels (Normal and stress), sub factor included zeolite application at two levels (with and without application) and biological fertilizers at four levels [1- Non application of mycorrhiza, 2-Application of mycorrhiza 3- Non -inoculation of bacteria 4- Inoculation of bacteria (
Pseudomonas, Azotobacter, Azospirillum
)] belonged to sub-sub factor. Analysis of variance results showed that interaction effect of zeolite and mycorrhiza and non-application of bacteria under normal irrigation regime on all measured traits (Instead potassium and nitrogen content) was significant. Mean comparison of treatments indicated that NZ
treatment (Zeolite and mycorrhiza application and non-inoculation of bacteria under normal irrigation regime) had highest amount of root yield (73340 kg.ha
), white sugar content (11.88%) and white sugar content (16.47%) but treatments of NM
(Mycorrhiza application and non-inoculation of bacteria under normal irrigation regime), DM
(Non-application of mycorrhiza and bacteria under stress irrigation regime) and DZ
(Non-application of zeolite, mycorrhiza and bacterial under stress irrigation regime) had highest amount of potassium content (4.45 meq.100g
sugar), amino-nitrogen (2.09 meq.100g
sugar) and sodium content (11.95 meq.100g
sugar), respectively. According to results of this research mycorrhiza inoculation and use of zeolite under water deficient conditions, caused improving sugar yield and consequently decreasing negative elements (Na, K, and N) under drought stress condition.
root yield
Sugar content
Abo-El-Goud, S. M. M. 2000.
Agronomic studies on fodder beet. Ph.D. Thesis. Faculty of Agriculture. Mansoura University. Egypt. 98 pp.
Amin, G. A., A. E. Badr. and M. H. M. Afifi. 2013.
Root Yield and quality of sugar beet (
Beta vulgaris
L.) in response to bio-fertilizer and foliar application with micronutrients. World App. Sci. J. 27(11): 1385-1389.
Brown, K. F, A. B. Messem, R. J. Dunham. and P. V. Biscoe. 1987.
Effect of drought on growth and water use of sugar beet. J. Agric. Sci. 109: 421-435.
Cattivelli, L., F. Rizza, F. W. Badeck, E. Mazzucotelli, A. M. Mastrangelo, E. Francia, C. Mare, A. Tondelli. and A. M. Stanca. 2008
. Drought tolerance improvement in crop plants: An integrated view from breeding to genomics. Field Crops Res. 105: 1-14.
Clover, G., H. Smith. and K. Jaggard. 1998.
The crop under stress. British Sugar Beet Review. 66(3): 17-19.
Clover, G. R. G., G. H. Smith, S. N. Azam-Ali. and K. W. Jaggard. 1999
. The effects of drought on sugar beet growth in isolation and in combination with beet yellows virus infection. J. Agri. Sci. Cambridge. 133: 251-261.
Cooke, D. and R. Scott. 1993.
The Sugar Beet Crop: Science into Practice. Chapman and Hill. New York. 98 pp.
Doubkova, P., E. Vlasakova. and R. Sudova. 2013.
Arbuscular mycorrhizal symbiosis alleviates drought stress imposed on
Knautia arvensis
plants in serpentine soil. Plant Soil. J. DOI: 10.1007/ s11104-013-1610-7. Online publication date: 1 Jan 2013.
El-Fouly, M. M., A. L. Rezk, M. A. Omer, E. E. Shalaby. and A. B. El-Nasharty. 2005.
Effect of potassium and foliar application of micronutrients on the yield and quality of sugar beet grown in calcareous soil. Egyptian J. Agric. Res. 2(2): 775-784.
El-Ghareib, E. A., M. A. El-Hawary, A. M. A. El-Shafai. and Y. E. E. El-Rayess. 2012.
Effect of Farmyard Manure, Plant Density and Bio-Fertilizer Treatments on Growth and Yield of Sugar Beet. J. Plant Production. 3(7): 2173-2187.
Farnia, A. and Gh. Hashemi. 2015.
Correlation between yield and other treats in sugar beet (
Beta vulgaris
L.) under application of different bio-fertilizers and irrigation. Int. J. Bio. Sci. 6(3): 146-152.
Favilli, F., R. Pastorelli. and A. Gori. 1993.
Response of sugar beet to Azospirillum bacterization in field experiments. Agric. Mediterr. J. 123: 281-285.
Ghareib, H. S. and A. S. El-Henawy. 2011.
Response of sugar beet (
Beta vulgaris
L) to irrigation regime, nitrogen rate and micronutrients application. Alex. Sci. Exchange J. 32: 140-156.
Hashemi, Gh., A. Farnia, M. Rahnamaeian. and M. Shaban. 2014.
Changes in carbohydrates and sugar yield in sugar beet (
Beta Vulgaris
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Ilyas, N., A. Bano. and S. Iqbal. 2008
. Variation in
strains isolated from maize growing in arid and semiarid areas. Inter. J. Agri. Biol.10: 612-8.
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Kavoosi, M. 2007
.Effects of zeolite application on rice yield, nitrogen recovery, and nitrogen use efficiency. Communications in Soil Science and Plant Analysis. England. 38(2): 69-76.
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. Drought stress induced changes in the biochemical parameters and photosynthetic pigments of cotton (
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. The search for drought tolerance in sugar beet. British Sugar Beet Review. J. 69(1): 40-43.
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. Physiological response of soybean (
Glycine max
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Payne, J. H .1968.
Sugar cane factory analytical control. The official methods of Hawaiian sugar technologists. 5
revised Ed.; produced by the Factory Methods Committee of the Hawaiian Sugar Technologists. Elsevier Publications. 63 pp.
Rassam, Gh., M. Dashti, A. R. Dadkhah. and A. Khoshnood Yazdi. 2015.
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Effect of drought stress on antioxidant enzymes activities with zeolite and selenium application in canola cultivars. Research on Crops. J. 12(2): 388-392.
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I came across an interesting quote in a newsletter from 2000 by a member of the University of Florida campus NOW chapter during a reelection campaign:
Feminism doesn't mean "be nice," I would like more open debate. All these appeals to sisterhood-just because you don't agree with somebody doesn't mean you don't like the person. It's a thing that we need to work out. "Be nice" - I don't like that idea. We want to be a united front but we also don't want to crush each other with everyone trying to be nice to everybody.


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Can Weather Pressure Changes Trigger Migraines?
For some people living with migraines, sudden weather changes and pressure can trigger an episode. Learn more about weather pressure and migraines here.
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The last time I touched alcohol I got a migraine so bad that I wanted to die. It was absolutely excruciating. I was on vacation and didn't know where an ER so I ended up...
Nightshades (i.e., tomatoes, peppers, potatoes, eggplant, etc.) are known for causing inflammation. Yet, I keep seeing them listed as part of diets that treat inflammation...
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Locate the cartilage in this section of the epiglottis. The ideally generalized form of connective tissue, with unspecialized proportions of the various matrix components and cells, receives its own name: The characteristics of the ground substance determine the permeability of the connective tissue layer to solutes and proteins.
Connective Tissue Lab
This heat-producing function is especially significant for infants and other small mammals and may be related to metabolic rate in adult humans. Never let this bland familiarity cause you to lose sight of the existence and significance of ordinary connective tissue.
1 2 34 5 Read more →


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You can draw features, such as buildings, water lines, tanks, pits and tracks on your farm map. To make sure the map isn't too cluttered you can
display just the features you need
for the job at hand.
Watch this video for an overview of how to draw features on the FarmIQ map:
For the PDF click here
In FarmIQ
features are managed in feature groups. So, for example, if you want to mark all your troughs on the map, you can add a feature group and call it Troughs, then in that feature group, you draw and name (or number) all the troughs at the correct locations.
There are some default feature groups at Farm level. Use these groups if you are logging effluent runs, recording pivot irrigation or recording weather and climate data against a particular weather station.These groups have special characteristics:
pivot irrigators.
Once the feature group has been added, most features, including effluent lines, can be drawn using the procedure below. Irrigators and weather stations, however, require additional input so please see Irrigators or Weather stations for more information.
If you are recording irrigation for pivot irrigators, you can draw the irrigator on the farm map.
In FarmIQ
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You can change the colour of features in a group, but not the feature shape. You cannot change the feature group name.
If you delete the last feature in a group, the feature group is also deleted. Click To delete a feature, below, for how to do this.
You can edit and delete features you have drawn.
When you edit a feature you can change its name and position.


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AGV Sport Echelon Gloves
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The top of the AGV Sport glove line, Echelon's leather palm and Amara reinforcements help with protection. Vents on the palm and fingers keep your hands cool, while titanium finger tips offer extra protection.
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Date published: 2017-09-17
Rated 1 out of 5 by Ivan from Fantastic value for the price !! Bought a size M, gloves look and feel great at first. In about an hour into my first ride, gloves started to really get uncomfortable and actually painful, around the knuckles area, this really started to get worse as I used and squeezed on the clutch. Unfortunately not a good fit for me, I had to return them which is a real shame, they do look great and feel soft and comfortable, except for the Knuckles area a deal breaker for me.
Date published: 2019-11-18
Rated 4 out of 5 by Shawn from Nice gloves Purchased these as my current gauntlets we're finally ready to be replaced. I didn't have the $400 this time around to buy them again. These looked like a good option for the price and they seem to be great so far. The fit and finish is actually really good. I don't however like the small finger attached to the ring as I use my two middle fingers for my levers. It was an easy task to cut it loose though with no harm to the glove. They run cooler than expected with the ginger ventilation but that's good to me. They're also quite stiff which is normal but a few quick cycles through the oven to form them to my hand and they're perfect. So far a great looking and functioning globe for a great price.
Date published: 2019-09-15
Rated 5 out of 5 by Savy from Simply the Best I love this product. Just perfect in every way. Better than the $250.00 Alpine Star I still have. The fit and the softness of the leather is unbeatable. Top quality protection. Cant beat this price.
Date published: 2019-09-15
Rated 5 out of 5 by Savy from Excellent Value Just recently got these gloves. Absolutely terrific product. The quality is fantastic. Better than gloves more than twice the price. They feel great. They are a touch big. The chart size for me would have been large. No large has ever fit me before. I read the other review posted that recommended getting the next size up because they fit small.I did order the size bigger. I am happy with the fit so decided to keep them. Best gloves I have ever purchased. Its like I having nothing on. That's the best fit.I got XL. Excellent comfort. Protection is outstanding. Quality is top grade. After 7 pairs finally got the ones that are perfect in ever way including price.
Date published: 2019-09-12
For the fitments, is the size chart in inches? According to this size chart, I'd be so small it isn't on the chart. But I wear large for Five and medium for Icon. So I'm wondering what size am I for AGV?
The sizing chart is in inches.
Date published: 2019-11-26
How waterproof are these gloves? I'm looking for something that will keep me decently warm in the rain.
I would not use these as dedicated rain gloves. If you are out and about and it starts raining, you would be fine for a short period of time. However, the space between the fingers is vented and will soak up water. As for warmth, these gloves are not insulated which means they will not hold in any heat if they are exposed to cold/wet conditions.
Date published: 2019-08-30
Are this gloves track ready? Looking for a pair for potential track day.
Great track gloves
Date published: 2019-10-03


text:
Don Perlin Returns to Transformers Art… Sort Of
Don Perlin is responsible for a great deal of the early Marvel comics, and is regarded as one of the originators of the Transformers aesthetic, at least on the page. He's been out of the Transformers loop for several years, until now. As a commission for longtime Friend of the Allspark Tim Finn, Don provided a really fantastic battle piece featuring Prime, Megatron, King Grimlock, as well as some of our favorite humans from the Marvel comics series, The Mechanic, Robot Master, and Circuit Breaker. What's the significance, you might ask? Well, Don's not drawn anything Transformers-related in nearly 18 years, and his work can be seen in IDW's current Transformers: Generations reprints. You can read Tim's thread and view the piece here.
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text:
The Guardian Reports on the authentication of fakes and the total lack of regulation in the art market.
A fake Marc Chagall painting, owned by a Leeds businessman who had bought it for £100,000 in 1992, was ordered to be burned last month and an Istanbul art gallery closed down its Joan Miró exhibition last year after directors of the Spanish surrealist painter's estate said some of the works were forgeries.
The uncovering of fakes by committees comprising descendants of the artist is increasingly common and has prompted one of Britain's foremost art historians to condemn the methods used by scholars to authenticate works as a "professional disgrace".
Martin Kemp, emeritus professor of art history at Oxford University and a Leonardo expert, said many rely on "dubious data" and that a "chaotic" approach is used to attribute paintings. He said he was alarmed at the ease with which historical, visual and scientific evidence is manipulated to suit the overriding objective – enhancing academic reputations or boosting financial rewards in attributing paintings to particular masters.
"There are no standards. It's totally unregulated. It's a Wild West … We're trying very hard to establish standards, but there is no professional control. You can shop around for a scientific report that probably says essentially what you want. It's shocking."
Nicholas Eastaugh, scientist and technical art historian of Art Access & Research.
Read The rest of the article over at The Guardian