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Sample records for cryptosporidium source tracking

  1. Cryptosporidium Source Tracking in the Potomac River Watershed

    EPA Science Inventory

    To better characterize the presence of Cryptosporidium in the Potomac River watershed, a PCR-based genotyping tool was used to analyze 64 base-flow and 28 storm-flow samples from five sites within the watershed. These sites included two water treatment plant intakes as well as t...

  2. Cryptosporidium source tracking in the Potomac River watershed - MCEARD

    EPA Science Inventory

    To better characterize Cryptosporidium in the Potomac River watershed, a PCR-based genotyping tool was used to analyze 64 base-flow and 28 storm-flow samples from five sites within the watershed. These sites included two water treatment plant intakes as well as three upstream si...

  3. Assessment of Giardia and Cryptosporidium spp. as a Microbial Source Tracking Tool for Surface Water: Application in a Mixed-Use Watershed

    PubMed Central

    Huck, Peter M.; Schreier, Hans; Isaac-Renton, Judith L.

    2014-01-01

    Knowledge of host specificity, combined with genomic sequencing of Giardia and Cryptosporidium spp., has demonstrated a microbial source tracking (MST) utility for these common waterborne microbes. To explore the source attribution potential of these pathogens, water samples were collected in a mixed rural-urban watershed in the Township of Langley, in southwestern British Columbia (BC), Canada, over a 2-year period. Cryptosporidium was detected in 63% of surface water samples at concentrations ranging from no positive detection (NPD) to 20,600 oocysts per 100 liters. Giardia was detected in 86% of surface water samples at concentrations ranging from NPD to 3,800 cysts per 100 liters of water. Sequencing at the 18S rRNA locus revealed that 50% of Cryptosporidium samples and 98% of Giardia samples contained species/genotypes (Cryptosporidium) or assemblages (Giardia) that are capable of infecting humans, based on current knowledge of host specificity and taxonomy. Cryptosporidium genotyping data were more promising for source tracking potential, due to the greater number of host-adapted (i.e., narrow-host-range) species/genotypes compared to Giardia, since 98% of Giardia isolates were zoonotic and the potential host could not be predicted. This report highlights the benefits of parasite genomic sequencing to complement Method 1623 (U.S. Environmental Protection Agency) and shows that Cryptosporidium subtyping for MST purposes is superior to the use of Giardia subtyping, based on better detection limits for Cryptosporidium-positive samples than for Giardia-positive samples and on greater host specificity among Cryptosporidium species. These additional tools could be used for risk assessment in public health and watershed management decisions. PMID:24463970

  4. Assessment of Giardia and Cryptosporidium spp. as a microbial source tracking tool for surface water: application in a mixed-use watershed.

    PubMed

    Prystajecky, Natalie; Huck, Peter M; Schreier, Hans; Isaac-Renton, Judith L

    2014-04-01

    Knowledge of host specificity, combined with genomic sequencing of Giardia and Cryptosporidium spp., has demonstrated a microbial source tracking (MST) utility for these common waterborne microbes. To explore the source attribution potential of these pathogens, water samples were collected in a mixed rural-urban watershed in the Township of Langley, in southwestern British Columbia (BC), Canada, over a 2-year period. Cryptosporidium was detected in 63% of surface water samples at concentrations ranging from no positive detection (NPD) to 20,600 oocysts per 100 liters. Giardia was detected in 86% of surface water samples at concentrations ranging from NPD to 3,800 cysts per 100 liters of water. Sequencing at the 18S rRNA locus revealed that 50% of Cryptosporidium samples and 98% of Giardia samples contained species/genotypes (Cryptosporidium) or assemblages (Giardia) that are capable of infecting humans, based on current knowledge of host specificity and taxonomy. Cryptosporidium genotyping data were more promising for source tracking potential, due to the greater number of host-adapted (i.e., narrow-host-range) species/genotypes compared to Giardia, since 98% of Giardia isolates were zoonotic and the potential host could not be predicted. This report highlights the benefits of parasite genomic sequencing to complement Method 1623 (U.S. Environmental Protection Agency) and shows that Cryptosporidium subtyping for MST purposes is superior to the use of Giardia subtyping, based on better detection limits for Cryptosporidium-positive samples than for Giardia-positive samples and on greater host specificity among Cryptosporidium species. These additional tools could be used for risk assessment in public health and watershed management decisions.

  5. Development and Evaluation of Three Real-Time PCR Assays for Genotyping and Source Tracking Cryptosporidium spp. in Water.

    PubMed

    Li, Na; Neumann, Norman F; Ruecker, Norma; Alderisio, Kerri A; Sturbaum, Gregory D; Villegas, Eric N; Chalmers, Rachel; Monis, Paul; Feng, Yaoyu; Xiao, Lihua

    2015-09-01

    The occurrence of Cryptosporidium oocysts in drinking source water can present a serious public health risk. To rapidly and effectively assess the source and human-infective potential of Cryptosporidium oocysts in water, sensitive detection and correct identification of oocysts to the species level (genotyping) are essential. In this study, we developed three real-time PCR genotyping assays, two targeting the small-subunit (SSU) rRNA gene (18S-LC1 and 18S-LC2 assays) and one targeting the 90-kDa heat shock protein (hsp90) gene (hsp90 assay), and evaluated the sensitivity and Cryptosporidium species detection range of these assays. Using fluorescence resonance energy transfer probes and melt curve analysis, the 18S-LC1 and hsp90 assays could differentiate common human-pathogenic species (C. parvum, C. hominis, and C. meleagridis), while the 18S-LC2 assay was able to differentiate nonpathogenic species (such as C. andersoni) from human-pathogenic ones commonly found in source water. In sensitivity evaluations, the 18S-LC2 and hsp90 genotyping assays could detect as few as 1 Cryptosporidium oocyst per sample. Thus, the 18S-LC2 and hsp90 genotyping assays might be used in environmental monitoring, whereas the 18S-LC1 genotyping assay could be useful for genotyping Cryptosporidium spp. in clinical specimens or wastewater samples.

  6. Source tracking identifies deer and geese as vectors of human-infectious Cryptosporidium genotypes in an urban/suburban watershed.

    PubMed

    Jellison, Kristen L; Lynch, Amy E; Ziemann, Joseph M

    2009-06-15

    This study identified Cryptosporidium genotypes in the Wissahickon watershed from May 2005 to April 2008. We analyzed 129 samples from Wissahickon Creek, 83 effluent samples from wastewater treatment plants (WWTPs), and 240 fecal droppings. Genotyping was based on the hypervariable region of the 18S rRNA gene. Oocysts were detected year-round, independent of wet weather events, in 22% of Wissahickon Creek samples, 5% of WWTP effluents, and 7% of fecal samples. Of the genotypes detected, 67% were human-infectious: 30% C. hominis or C. hominis-like, 12% C. parvum, 14% cervine genotype, 9% skunk genotype, and 1% chipmunk I genotype. Similar genotype profiles were detected in Wissahickon Creek each year, and human-infectious genotypes were detected year-round. Unusual genotypes detected in a deer (a C. hominis-like genotype) and geese (C. hominis-like genotypes, C. parvum, and muskrat genotype I) show that these animals are vectors of human-infectious genotypes in this watershed. Results suggest that deer, geese, and WWTPs are appropriate targets for source water protection in the Wissahickon watershed.

  7. GENOTYPING CRYPTOSPORIDIUM OOCYSTS IN WATER SAMPLES AS A TOOL FOR THE IDENTIFICATION OF CONTAMINATION SOURCES

    EPA Science Inventory

    The identification of Cryptosporidium oocysts in environmental samples is largely made by the use of an immunofluorescent assay (IFA). Because IFA detects oocysts from all Cryptosporidium parasites, the species distribution and source of Cryptosporidium parasites in environmental...

  8. ANIMAL SOURCE IDENTIFICATION USING A CRYPTOSPORIDIUM DNA CHARACTERIZATION TECHNIQUE

    EPA Science Inventory

    This document summarizes the application of a particular molecular method to improve detection and differentiation of species and genotypes of Cryptosporidium oocysts found in environmental samples. Of particular interest is the method's potential for determining the source anim...

  9. CRYPTOSPORIDIUM SOURCE TRACKING TO ENHANCE SOURCE WATER PROTECTION IMPLEMENTATION IN THE POTOMAC RIVER WATERSHED: A REGIONAL APPLIED RESEARCH EFFORTS (RARE) PROJECT

    EPA Science Inventory

    The Potomac River watershed is a critical drinking water supply for the Washington DC metropolitan area. In 2004, the Drinking Water Source Protection Partnership (DWSPP) was formed to help coordinate efforts by local drinking water utilities and government agencies to protect th...

  10. DETECTION OF CRYPTOSPORIDIUM OOCYSTS IN SOURCE AND FINISHED WATERS

    EPA Science Inventory

    Numerous waterborne outbreaks of cryptosporidiosis have occurred with the most notable being the 1993 episode in Milwaukee. As a result, the past decade has seen a massive effort expended on the development of methods to detect Cryptosporidium parvum oocysts in source and finish...

  11. IDENTIFICATION OF SPECIES AND SOURCES OF CRYPTOSPORIDIUM OOCYSTS IN STORM WATERS BY A SMALL SUBUNIT RRNA-BASED DIAGNOSTIC AND GENOTYPING TOOL

    EPA Science Inventory

    The identification of Cryptosporidium oocysts in environmental samples is largely made by the use of immunofluorescent assay (IFA). because IFA detects oocysts from all Cryptosporidium parasites, the species distribution and source of Cryptosporidium parasites in environmental sa...

  12. IDENTIFICATION OF CRYPTOSPORIDIUM SPECIES AND SOURCES IN RAW WASTEWATER USING A SMALL SUBUNIT RRNA-BASED PCR-RFLP TOOL

    EPA Science Inventory

    The species composition and source of Cryptosporidium oocysts in wastewater have never been determined, even though it is widely assumed that these oocysts are from human sewage. Recent molecular characterizations of Cryptosporidium parasites make it possible to differentiate hum...

  13. Application of leftover sample material from waterborne protozoa monitoring for the molecular detection of Bacteroidales and fecal source tracking markers

    EPA Science Inventory

    In this study, we examined the potential for detecting fecal bacteria and microbial source tracking markers in samples discarded during the concentration of Cryptosporidium and Giardia using USEPA Method 1623. Recovery rates for different fecal bacteria were determined using sp...

  14. SEROLOGICAL RESPONSES TO CRYPTOSPORIDIUM ANTIGENS AMONG USERS OF SURFACE VERSUS UNDERGROUND DRINKING WATER SOURCES

    EPA Science Inventory

    Cryptosporidium oocysts have been detected in source and treated drinking waters in the United States and elsewhere. Enhanced enteric disease surveillance, initiated following detection of oocysts, has not often detected elevated rates of infection or of symptoms compatible with...

  15. Cryptosporidium: Treatment

    MedlinePlus

    ... Cryptosporidium Tracking in the United States CryptoNet Healthy Water Sites Healthy Water Drinking Water Healthy Swimming Global ... and HIV-Infected Children [PDF - 384 pages] Healthy Water Sites Healthy Water Drinking Water Healthy Swimming Global ...

  16. Changes in Escherichia coli to Cryptosporidium ratios for various fecal pollution sources and drinking water intakes.

    PubMed

    Lalancette, Cindy; Papineau, Isabelle; Payment, Pierre; Dorner, Sarah; Servais, Pierre; Barbeau, Benoit; Di Giovanni, George D; Prévost, Michèle

    2014-05-15

    Assessing the presence of human pathogenic Cryptosporidium oocysts in surface water remains a significant water treatment and public health challenge. Most drinking water suppliers rely on fecal indicators, such as the well-established Escherichia coli (E. coli), to avoid costly Cryptosporidium assays. However, the use of E. coli has significant limitations in predicting the concentration, the removal and the transport of Cryptosporidium. This study presents a meta-analysis of E. coli to Cryptosporidium concentration paired ratios to compare their complex relationships in eight municipal wastewater sources, five agricultural fecal pollution sources and at 13 drinking water intakes (DWI) to a risk threshold based on US Environmental Protection Agency (USEPA) regulations. Ratios lower than the USEPA risk threshold suggested higher concentrations of oocysts in relation to E. coli concentrations, revealing an underestimed risk for Cryptosporidium based on E. coli measurements. In raw sewage (RS), high ratios proved E. coli (or fecal coliforms) concentrations were a conservative indicator of Cryptosporidium concentrations, which was also typically true for secondary treated wastewater (TWW). Removals of fecal indicator bacteria (FIB) and parasites were quantified in WWTPs and their differences are put forward as a plausible explanation of the sporadic ratio shift. Ratios measured from agricultural runoff surface water were typically lower than the USEPA risk threshold and within the range of risk misinterpretation. Indeed, heavy precipitation events in the agricultural watershed led to high oocyst concentrations but not to E. coli or enterococci concentrations. More importantly, ratios established in variously impacted DWI from 13 Canadian drinking water plants were found to be related to dominant fecal pollution sources, namely municipal sewage. In most cases, when DWIs were mainly influenced by municipal sewage, E. coli or fecal coliforms concentrations agreed with

  17. 40 CFR 141.211 - Special notice for repeated failure to conduct monitoring of the source water for Cryptosporidium...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... determine whether water treatment at the (treatment plant name) is sufficient to adequately remove... source of your drinking water for Cryptosporidium in order to determine by (date) whether water treatment... conduct monitoring of the source water for Cryptosporidium and for failure to determine bin...

  18. 40 CFR 141.211 - Special notice for repeated failure to conduct monitoring of the source water for Cryptosporidium...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... determine whether water treatment at the (treatment plant name) is sufficient to adequately remove... source of your drinking water for Cryptosporidium in order to determine by (date) whether water treatment... conduct monitoring of the source water for Cryptosporidium and for failure to determine bin...

  19. 40 CFR 141.211 - Special notice for repeated failure to conduct monitoring of the source water for Cryptosporidium...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... determine whether water treatment at the (treatment plant name) is sufficient to adequately remove... source of your drinking water for Cryptosporidium in order to determine by (date) whether water treatment... conduct monitoring of the source water for Cryptosporidium and for failure to determine bin...

  20. 40 CFR 141.211 - Special notice for repeated failure to conduct monitoring of the source water for Cryptosporidium...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... determine whether water treatment at the (treatment plant name) is sufficient to adequately remove... source of your drinking water for Cryptosporidium in order to determine by (date) whether water treatment... conduct monitoring of the source water for Cryptosporidium and for failure to determine bin...

  1. 40 CFR 141.211 - Special notice for repeated failure to conduct monitoring of the source water for Cryptosporidium...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... determine whether water treatment at the (treatment plant name) is sufficient to adequately remove... source of your drinking water for Cryptosporidium in order to determine by (date) whether water treatment... conduct monitoring of the source water for Cryptosporidium and for failure to determine bin...

  2. Investigating source water Cryptosporidium concentration, species and infectivity rates during rainfall-runoff in a multi-use catchment.

    PubMed

    Swaffer, Brooke A; Vial, Hayley M; King, Brendon J; Daly, Robert; Frizenschaf, Jacqueline; Monis, Paul T

    2014-12-15

    Protozoan pathogens present a significant human health concern, and prevention of contamination into potable networks remains a key focus for drinking water providers. Here, we monitored the change in Cryptosporidium concentration in source water during high flow events in a multi-use catchment. Furthermore, we investigated the diversity of Cryptosporidium species/genotypes present in the source water, and delivered an oocyst infectivity fraction. There was a positive and significant correlation between Cryptosporidium concentration and flow (ρ = 0.756) and turbidity (ρ = 0.631) for all rainfall-runoff events, despite variable source water pathogen concentrations. Cell culture assays measured oocyst infectivity and suggested an overall source water infectious fraction of 3.1%. No infectious Cryptosporidium parvum or Cryptosporidium hominis were detected, although molecular testing detected C. parvum in 7% of the samples analysed using PCR-based molecular techniques. Twelve Cryptosporidium species/genotypes were identified using molecular techniques, and were reflective of the host animals typically found in remnant vegetation and agricultural areas. The inclusion of molecular approaches to identify Cryptosporidium species and genotypes highlighted the diversity of pathogens in water, which originated from various sources across the catchment. We suggest this mixing of runoff water from a range of landuses containing diverse Cryptosporidium hosts is a key explanation for the often-cited difficulty forming strong pathogen-indicator relationships.

  3. IDENTIFICATION OF CRYPTOSPORIDIUM SPECIES AND THE SOURCES IN RAW WASTEWATER USING A SMALL SUBUNIT RRNA-BASED PCR-RFLP TOOL

    EPA Science Inventory

    The species composition and source of Cryptosporidium oocysts in wastewater have never been determined, even though it is widely assumed that these oocysts are from human sewage. Recent molecular characterizations of Cryptosporidium parasites make it possible to differentiate hum...

  4. Prevalence of Cryptosporidium species in recreational versus non-recreational water sources.

    PubMed

    Loganthan, Sasdekumar; Yang, Rongchang; Bath, Andrew; Gordon, Cameron; Ryan, Una

    2012-08-01

    Cryptosporidiosis, caused by the protozoan parasite Cryptosporidium, represents the major public health concern of water utilities in developed nations due to its small size, resistance to disinfection and ability to be shed in large numbers in faeces. In Australia, recreational access is not allowed on direct supply sources, however, in Western Australia, limited recreational access to drinking water catchments has been allowed, although only in the outer catchment. Recreational activities within 2 km of the drinking water body is prohibited. The present study compared the amount, prevalence and species of Cryptosporidium in recreational versus non-recreational water catchments in the south west of Western Australia (WA). Recreational water catchments, which allowed swimming and camping had a higher prevalence of Cryptosporidium and the majority of samples were the human-associated C. hominis. Non-recreational catchments had a lower prevalence and all the samples genotyped were C. parvum. Risk analysis identified increasing population as strongly correlated with an increase in the prevalence of Cryptosporidium in recreational catchments. This suggests that recreational access to drinking water catchments is a serious public health risk and government policy limiting activities to the outer catchment should be supported.

  5. MICROBIAL SOURCE TRACKING

    EPA Science Inventory

    Fecal contamination of waters used for recreation, drinking water, and aquaculture is an environmental problem and poses significant human health risks. The problem is often difficult to correct because the source of the contamination cannot be determined with certainty. Run-of...

  6. Sources and Species of Cryptosporidium Oocysts in the Wachusett Reservoir Watershed

    PubMed Central

    Jellison, Kristen L.; Hemond, Harold F.; Schauer, David B.

    2002-01-01

    Understanding the behavior of Cryptosporidium oocysts in the environment is critical to developing improved watershed management practices for protection of the public from waterborne cryptosporidiosis. Analytical methods of improved specificity and sensitivity are essential to this task. We developed a nested PCR-restriction fragment length polymorphism assay that allows detection of a single oocyst in environmental samples and differentiates the human pathogen Cryptosporidium parvum from other Cryptosporidium species. We tested our method on surface water and animal fecal samples from the Wachusett Reservoir watershed in central Massachusetts. We also directly compared results from our method with those from the immunofluorescence microscopy assay recommended in the Information Collection Rule. Our results suggest that immunofluorescence microscopy may not be a reliable indicator of public health risk for waterborne cryptosporidiosis. Molecular and environmental data identify both wildlife and dairy farms as sources of oocysts in the watershed, implicate times of cold water temperatures as high-risk periods for oocyst contamination of surface waters, and suggest that not all oocysts in the environment pose a threat to public health. PMID:11823192

  7. Serological responses to Cryptosporidium antigens among users of surface- vs. ground-water sources.

    PubMed Central

    Frost, F. J.; Kunde, T. R.; Muller, T. B.; Craun, G. F.; Katz, L. M.; Hibbard, A. J.; Calderon, R. L.

    2003-01-01

    Cryptosporidium oocysts are commonly detected in surface-derived drinking water. However, the public health significance of these findings is unclear. This study compared serological responses to two Cryptosporidium antigen groups for blood donors and college students using chlorinated and filtered river water vs. ground-water sources. The surface water received agricultural and domestic sewage discharges upstream. Participants from the surface-water city had a higher relative prevalence (RP) of a serological response to the 15/17-kDa antigen group (72.3 vs. 52.4%, RP = 1.36, P < 0.001) and to the 27-kDa antigen group (82.6 vs. 72.5%, RP = 1.14, P < 0.02). Multivariate logistic regression analysis found that the people with a shorter duration of residence or drinking bottled water also had a lower seropositivity for each marker. Use of private wells was associated with a higher prevalence of response to the 15/17-kDa markers. Seroconversion to the 15/17-kDa antigen group was more common in the residents of the city using surface water. These findings are consistent with an increased risk of Cryptosporidium infection for users of surface-derived drinking water compared with users of municipal ground-water-derived drinking water. Users of private well water may also have an increased risk of infection. PMID:14959781

  8. Cryptosporidium: Infection - Immunocompromised Persons

    MedlinePlus

    ... Cryptosporidium Tracking in the United States CryptoNet Healthy Water Sites Healthy Water Drinking Water Healthy Swimming Global ... getting cryptosporidiosis? See Prevention & Control - Immunocompromised Persons. Healthy Water Sites Healthy Water Drinking Water Healthy Swimming Global ...

  9. Cryptosporidium: Prevention - Immunocompromised Persons

    MedlinePlus

    ... Cryptosporidium Tracking in the United States CryptoNet Healthy Water Sites Healthy Water Drinking Water Healthy Swimming Global ... diarrhea, have it tested for Crypto. Avoid swallowing water when swimming in the ocean, lakes, rivers, or ...

  10. Occurrence, source, and human infection potential of Cryptosporidium and Enterocytozoon bieneusi in drinking source water in Shanghai, China, during a pig carcass disposal incident.

    PubMed

    Hu, Yue; Feng, Yaoyu; Huang, Chengchen; Xiao, Lihua

    2014-12-16

    In March 2013, thousands of domestic pig carcasses were found floating in the Huangpu River, a drinking source water in Shanghai, China. To investigate the impact of the pig carcass incident on microbial water quality, 178 river water samples were collected from the upper Huangpu River from March 2013 to March 2014. Samples were concentrated by calcium carbonate flocculation and examined for host-adapted Cryptosporidium and Enterocytozoon bieneusi by ploymerase chain reaction (PCR). Positive PCR products were sequenced to determine Cryptosporidium species and E. bieneusi genotypes. A total of 67 (37.6%) and 56 (31.5%) samples were PCR-positive for Cryptosporidium and E. bieneusi, respectively. The occurrence rates of Cryptosporidium and E. bieneusi in March 2013 (83.3%; 41.7%) and May 2013 (73.5%; 44.1%) were significantly higher than rates in later sampling times. Among the 13 Cryptosporidium species/genotypes identified, C. andersoni and C. suis were the most common species, being found in 38 and 27 samples, respectively. Seventeen E. bieneusi genotypes were found, belonging to 11 established genotypes (EbpC, EbpA, D, CS-8, PtEb IX, Peru 8, Peru 11, PigEBITS4, EbpB, G, O) and six new ones (RWSH1 to RWSH6), most of which belonged to pig-adapted Groups 1d and 1e. EbpC was the most common genotype, being found in 37 samples. The distribution of Cryptosporidium species and E. bieneusi genotypes suggest that dead pigs contributed significantly to Cryptosporidium and E. bieneusi contamination in the Huangpu River. Although most Cryptosporidium species found in river water were not major human pathogens, the majority of E. bieneusi genotypes detected were endemic in China. Data from this study should be useful in the development of strategies in addressing future contamination events in drinking water supplies.

  11. Identifying host sources, human health risk and indicators of Cryptosporidium and Giardia in a Canadian watershed influenced by urban and rural activities.

    PubMed

    Van Dyke, Michele I; Ong, Corinne S L; Prystajecky, Natalie A; Isaac-Renton, Judith L; Huck, Peter M

    2012-06-01

    Cryptosporidium and Giardia were characterized in a watershed in southern Ontario, Canada, over a 2½ year period. River samples were collected every two weeks, primarily near a municipal drinking water treatment plant intake. Cryptosporidium and Giardia were frequently detected with an overall occurrence rate of 88 and 97%, respectively. Giardia concentrations were higher than Cryptosporidium, with median values of 80 cysts 100 L(-1) and 12 oocysts 100 L(-1), respectively. Although pathogens rarely show a significant relationship with fecal or water quality indicators, this study determined that Cryptosporidium, but not Giardia, was significantly correlated with Escherichia coli, turbidity and river flow. There was no correlation between the two types of protozoa, and only Giardia showed a seasonal trend with higher concentrations at cold water temperatures. Cryptosporidium genotyping of all samples found that farm animals and wildlife were an important contributor of oocysts in the watershed, and that Cryptosporidium strains/genotypes of medium to high risk for human infection (C. hominis, C. parvum and C. ubiquitum) were detected in 16% of samples. This study was able to identify Cryptosporidium host sources and human health risk, and to identify differences between Cryptosporidium and Giardia occurrence in the watershed.

  12. Occurrence of Cryptosporidium and Giardia on beef farms and water sources within the vicinity of the farms on Prince Edward Island, Canada.

    PubMed

    Budu-Amoako, Ebo; Greenwood, Spencer J; Dixon, Brent R; Barkema, Herman W; McClure, J T

    2012-02-28

    The objectives of this study were to determine the prevalence and assemblages of Giardia and species of Cryptosporidium on beef farms in Prince Edward Island (PEI), Canada, including the water sources associated with the farms, and to determine risk factors for infection of cattle with these parasites. Twenty beef farms were selected based on the presence of surface water< 500 m from the barn. Prevalence was determined by direct immunofluorescence microscopy, while genotyping and species determination were performed by nested-PCR and DNA sequencing. Giardia was detected in 42% (95% CI: 38-46%) of fecal samples from 100% farms while Cryptosporidium was detected in 17% (95% CI: 14-19%) of fecal samples from 80% of farms. The most predominant Giardia assemblage isolated was the livestock specific assemblage E (89%). The zoonotic assemblages A and B were found in 4 and 7% of the Giardia isolates that were genotyped, respectively. The Giardia assemblages were detected equally between the cows and calves examined. Overall, the most common Cryptosporidium species detected in this study was Cryptosporidium andersoni (49%), predominantly found in cattle > 6 mo of age, while most Cryptosporidium bovis and Cryptosporidium pestis (previously Cryptosporidium parvum 'bovine genotype') isolates were detected in calves ≤ 6 mo of age. All Cryptosporidium ryanae isolates (four) were found in calves. Giardia cysts and Cryptosporidium oocysts were detected in 14 and 93% of surface water samples of 14 farms, respectively. Cryptosporidium oocysts were detected in three (15%) ground water samples of 20 farms. One Cryptosporidium-positive water sample, which was the only surface water sample amenable to genotyping, contained C. parvum. The farm-level risk factors investigated in this study, age of animals and location of the farm, were not associated with the risk of infection in cattle with either Cryptosporidium spp. or Giardia duodenalis. We conclude that beef cattle are a potential

  13. THE FUTURE OF MICROBIAL SOURCE TRACKING STUDIES

    EPA Science Inventory

    Microbial source tracking (MST) is differentiated from traditional microbial water quality efforts by the need to identify the host species from which the bacteria originate, rather than necessarily identifying an individual point source. Despite recent advances in the developmen...

  14. Estimation of pathogen concentrations in a drinking water source using hydrodynamic modelling and microbial source tracking.

    PubMed

    Sokolova, Ekaterina; Aström, Johan; Pettersson, Thomas J R; Bergstedt, Olof; Hermansson, Malte

    2012-09-01

    The faecal contamination of drinking water sources can lead to waterborne disease outbreaks. To estimate a potential risk for waterborne infections caused by faecal contamination of drinking water sources, knowledge of the pathogen concentrations in raw water is required. We suggest a novel approach to estimate pathogen concentrations in a drinking water source by using microbial source tracking data and fate and transport modelling. First, the pathogen (norovirus, Cryptosporidium, Escherichia coli O157/H7) concentrations in faecal contamination sources around the drinking water source Lake Rådasjön in Sweden were estimated for endemic and epidemic conditions using measured concentrations of faecal indicators (E. coli and Bacteroidales genetic markers). Afterwards, the fate and transport of pathogens within the lake were simulated using a three-dimensional coupled hydrodynamic and microbiological model. This approach provided information on the contribution from different contamination sources to the pathogen concentrations at the water intake of a drinking water treatment plant. This approach addresses the limitations of monitoring and provides data for quantitative microbial risk assessment (QMRA) and risk management in the context of faecal contamination of surface drinking water sources.

  15. Cryptosporidium and Giardia as foodborne zoonoses.

    PubMed

    Smith, H V; Cacciò, S M; Cook, N; Nichols, R A B; Tait, A

    2007-10-21

    Cryptosporidium and Giardia are major causes of diarrhoeal disease in humans, worldwide and are major causes of protozoan waterborne diseases. Both Cryptosporidium and Giardia have life cycles which are suited to waterborne and foodborne transmission. There are 16 'valid'Cryptosporidium species and a further 33+ genotypes described. Parasites which infect humans belong to the Giardia duodenalis "type", and at least seven G. duodenalis assemblages are recognised. Cryptosporidium parvum is the major zoonotic Cryptosporidium species, while G. duodenalis assemblages A and B have been found in humans and most mammalian orders. In depth studies to determine the role of non-human hosts in the transmission of Cryptosporidium and Giardia to humans are required. The use of harmonised methodology and standardised and validated molecular markers, together with sampling strategies that provide sufficient information about all contributors to the environmental (oo)cyst pool that cause contamination of food and water, are recommended. Standardised methods for detecting (oo)cysts in water are available, as are optimised, validated methods for detecting Cryptosporidium in soft fruit and salad vegetables. These provide valuable data on (oo)cyst occurrence, and can be used for species and subspecies typing using appropriate molecular tools. Given the zoonotic potential of these organisms, epidemiological, source and disease tracking investigations involve multidisciplinary teams. Here, the role of the veterinarian is paramount, particularly in understanding the requirement for adopting comprehensive sampling strategies for analysing both sporadic and outbreak samples from all potential non-human contributors. Comprehensive sampling strategies increase our understanding of parasite population biology and structure and this knowledge can be used to determine what level of discrimination is required between isolates. Genetic exchange is frequent in C. parvum populations, leading to

  16. Cryptosporidium (Crypto) Disease: Diagnosis & Detection

    MedlinePlus

    ... Cryptosporidium Tracking in the United States CryptoNet Healthy Water Sites Healthy Water Drinking Water Healthy Swimming Global ... Cryptosporidiosis [DPDx] Diagnostic Procedures: Stool Specimens [DPDx] Healthy Water Sites Healthy Water Drinking Water Healthy Swimming Global ...

  17. Modeling Cryptosporidium and Giardia in Ground and Surface Water Sources in Rural India: Associations with Latrines, Livestock, Damaged Wells, and Rainfall Patterns.

    PubMed

    Daniels, Miles E; Smith, Woutrina A; Schmidt, Wolf-Peter; Clasen, Thomas; Jenkins, Marion W

    2016-07-19

    Surface and groundwater contamination with fecal pathogens is a public health concern especially in low-income settings where these sources are used untreated. We modeled observed Cryptosporidium and Giardia contamination in community ponds (n = 94; 79% contaminated), deep tubewells (DTWs) (n = 107; 17%), and shallow tubewells (STWs) (n = 96; 19%) during the 2012 and 2013 monsoon seasons (June-August) in 60 villages in Puri District, India to understand sources and processes of contamination. Detection of Cryptosporidium and/or Giardia in a tubewell was positively associated with damage to the well pad for DTWs, the amount of human loading into pour-flush latrine pits nearby (≤15 m) for STWs, and the village literacy rate (for Giardia in STWs). Pond concentration levels were positively associated with the number of people practicing open defecation within 50 m and the sheep population for Cryptosporidium, and with the village illiteracy rate for Giardia. Recent rainfall increased the risk of Cryptosporidium in STWs (an extreme event) and ponds (any), while increasing seasonal rainfall decreased the risk of Giardia in STWs and ponds. Full latrine coverage in this setting is expected to marginally reduce pond Cryptosporidium contamination (16%) while increasing local groundwater protozoal contamination (87-306%), with the largest increases predicted for Cryptosporidium in STWs.

  18. Modeling Cryptosporidium and Giardia in Ground and Surface Water Sources in Rural India: Associations with Latrines, Livestock, Damaged Wells, and Rainfall Patterns

    PubMed Central

    2016-01-01

    Surface and groundwater contamination with fecal pathogens is a public health concern especially in low-income settings where these sources are used untreated. We modeled observed Cryptosporidium and Giardia contamination in community ponds (n = 94; 79% contaminated), deep tubewells (DTWs) (n = 107; 17%), and shallow tubewells (STWs) (n = 96; 19%) during the 2012 and 2013 monsoon seasons (June–August) in 60 villages in Puri District, India to understand sources and processes of contamination. Detection of Cryptosporidium and/or Giardia in a tubewell was positively associated with damage to the well pad for DTWs, the amount of human loading into pour-flush latrine pits nearby (≤15 m) for STWs, and the village literacy rate (for Giardia in STWs). Pond concentration levels were positively associated with the number of people practicing open defecation within 50 m and the sheep population for Cryptosporidium, and with the village illiteracy rate for Giardia. Recent rainfall increased the risk of Cryptosporidium in STWs (an extreme event) and ponds (any), while increasing seasonal rainfall decreased the risk of Giardia in STWs and ponds. Full latrine coverage in this setting is expected to marginally reduce pond Cryptosporidium contamination (16%) while increasing local groundwater protozoal contamination (87–306%), with the largest increases predicted for Cryptosporidium in STWs. PMID:27310009

  19. MICROBIAL SOURCE TRACKING: DIFFERENT USES AND APPROACHES

    EPA Science Inventory

    Microbial Source Tracking (MST) methods are used to determine the origin of fecal pollution impacting natural water systems. Several methods require the isolation of pure cultures in order to develop phenotypic or genotypic fingerprint libraries of both source and water bacterial...

  20. Cryptosporidium enteritis

    MedlinePlus

    Proper sanitation and hygiene, including handwashing, are important measures for preventing this illness. Certain water filters can also reduce risk by filtering out the cryptosporidium eggs. However, ...

  1. MICROBIAL SOURCE TRACKING GUIDE DOCUMENT

    EPA Science Inventory

    Approximately 13% of surface waters in the United States do not meet designated use criteria as determined by high densities of fecal indicator bacteria. Although some of the contamination is attributed to point sources such as confined animal feeding operation (CAFO) and wastew...

  2. Foodborne illness associated with Cryptosporidium and Giardia from livestock.

    PubMed

    Budu-Amoako, Ebo; Greenwood, Spencer J; Dixon, Brent R; Barkema, Herman W; McClure, J T

    2011-11-01

    Waterborne outbreaks caused by Cryptosporidium and Giardia are well documented, while the public health implications for foodborne illness from these parasites have not been adequately considered. Cryptosporidium and Giardia are common in domestic livestock, where young animals can have a high prevalence of infection, shedding large numbers of oocysts and cysts. Molecular epidemiological studies have advanced our knowledge on the distribution of Cryptosporidium and Giardia species and genotypes in specific livestock. This has enabled better source tracking of contaminated foods. Livestock generate large volumes of fecal waste, which can contaminate the environment with (oo)cysts. Evidence suggests that livestock, particularly cattle, play a significant role in food contamination, leading to outbreaks of cryptosporidiosis. However, foodborne giardiasis seems to originate primarily from anthroponotic sources. Foodborne cryptosporidiosis and giardiasis are underreported because of the limited knowledge of the zoonotic potential and public health implications. Methods more sensitive and cheaper are needed to detect the often-low numbers of (oo)cysts in contaminated food and water. As the environmental burden of Cryptosporidium oocysts and Giardia cysts from livestock waste increases with the projected increase in animal agriculture, public health is further compromised. Contamination of food by livestock feces containing Cryptosporidium oocysts and Giardia cysts could occur via routes that span the entire food production continuum. Intervention strategies aimed at preventing food contamination with Cryptosporidium and Giardia will require an integrated approach based on knowledge of the potential points of entry for these parasites into the food chain. This review examines the potential for foodborne illness from Cryptosporidium and Giardia from livestock sources and discusses possible mechanisms for prevention and control.

  3. Cryptosporidium and Giardia in Humans, Domestic Animals, and Village Water Sources in Rural India

    PubMed Central

    Daniels, Miles E.; Shrivastava, Arpit; Smith, Woutrina A.; Sahu, Priyadarshi; Odagiri, Mitsunori; Misra, Pravas R.; Panigrahi, Pinaki; Suar, Mrutyunjay; Clasen, Thomas; Jenkins, Marion W.

    2015-01-01

    Cryptosporidium parvum and Giardia lamblia are zoonotic enteric protozoa of significant health concern where sanitation, hygiene, and water supplies are inadequate. We examined 85 stool samples from diarrhea patients, 111 pooled fecal samples by species across seven domestic animal types, and water from tube wells (N = 207) and ponds (N = 94) across 60 villages in coastal Odisha, India, for Cryptosporidium oocysts and Giardia cysts to measure occurrence, concentration/shedding, and environmental loading rates. Oocysts/cysts were detected in 12% of diarrhea patients. Detection ranged from 0% to 35% for Cryptosporidium and 0% to 67% for Giardia across animal hosts. Animal loading estimates indicate the greatest contributors of environmental oocysts/cysts in the study region are cattle. Ponds were contaminated with both protozoa (oocysts: 37%, cysts: 74%), as were tube wells (oocysts: 10%, cysts: 14%). Future research should address the public health concern highlighted from these findings and investigate the role of domestic animals in diarrheal disease transmission in this and similar settings. PMID:26123963

  4. Transport of Cryptosporidium, Giardia, Source-specific Indicator Organisms, and Standard Water Quality Constituents During Storm Events

    NASA Astrophysics Data System (ADS)

    Sturdevant-Rees, P. L.; Bourdeau, D.; Baker, R.; Long, S. C.; Barten, P. K.

    2004-05-01

    Microbial and water-quality measurements are collected during storm events under a variety of meteorological and land-use conditions in order to 1) identify risk of Cryptosporidium oocysts, Giardia cysts and other constituents, including microbial indicator organisms, entering surface waters from various land uses during periods of surface runoff; 2) optimize storm sampling procedures for these parameters; and 3) optimize strategies for accurate determination of constituent loads. The investigation is focused on four isolated land uses: forested with free ranging wildlife, beaver influenced forested with free ranging wildlife, residential/commercial, and dairy farm grazing/pastureland using an upstream and downstream sampling strategy. Traditional water-quality analyses include pH, temperature, turbidity, conductivity, total suspended solids, total phosphorus, total Kjeldahl-nitrogen, and ammonia nitrogen, Giardia cysts and Cryptosporidium oocysts. Total coliforms and fecal coliforms are measured as industry standard microbial analyses. Sorbitol-fermenting Bifidobacteria, Rhodococcus coprophilus, Clostridium perfringens spores, and Somatic and F-specific coliphages are measured at select sites as potential alternative source-specific indicator organisms. Upon completion of the project, the final database will consist of wet weather transport data for a set of parameters during twenty-four distinct storm-events in addition to monthly baseline data. A subset of the results to date will be presented, with focus placed on demonstrating the impact of beaver on constituent loadings over a variety of hydrologic and meteorological conditions.

  5. Source water assessment and nonpoint sources of acutely toxic contaminants: A review of research related to survival and transport of Cryptosporidium parvum

    NASA Astrophysics Data System (ADS)

    Walker, Mark J.; Montemagno, Carlo D.; Jenkins, Michael B.

    1998-12-01

    Amendments to the Safe Drinking Water Act (PL-930123) in 1996 required that public water supply managers identify potential sources of contamination within contributing areas. Nonpoint sources of acutely toxic microbial contaminants, such as Cryptosporidium parvum, challenge current approaches to source identification and management as a first step toward developing management plans for public water supply protection. Little may be known about survival and transport in the field environment, prescribed practices may not be designed to manage such substances, and infective stages may be present in vast numbers and may resist water treatment and disinfection processes. This review summarizes research related to survival and transport of C. parvum oocysts, as an example of an acutely toxic contaminant with nonpoint sources in animal agriculture. It discusses ∥1) significance of infected domesticated animals as potential sources of C. parvum, (2) laboratory and field studies of survival and transport, and (3) approaches to source control in the context of public health protection.

  6. Seasonal variation and potential sources of Cryptosporidium contamination in surface waters of Chao Phraya River and Bang Pu Nature Reserve pier, Thailand.

    PubMed

    Koompapong, Khuanchai; Sukthana, Yaowalark

    2012-07-01

    Using molecular techniques, a longitudinal study was conducted with the aims at identifying the seasonal difference of Cryptosporidium contamination in surface water as well as analyzing the potential sources based on species information. One hundred forty-four water samples were collected, 72 samples from the Chao Phraya River, Thailand, collected in the summer, rainy and cool seasons and 72 samples from sea water at Bang Pu Nature Reserve pier, collected before, during and after the presence of migratory seagulls. Total prevalence of Cryptosporidium contamination in river and sea water locations was 11% and 6%, respectively. The highest prevalence was observed at the end of rainy season continuing into the cool season in river water (29%) and in sea water (12%). During the rainy season, prevalence of Cryptosporidium was 4% in river and sea water samples, but none in summer season. All positive samples from the river was C. parvum, while C. meleagridis (1), and C. serpentis (1) were obtained from sea water. To the best of our knowledge, this is the first genetic study in Thailand of Cryptosporidium spp contamination in river and sea water locations and the first report of C. serpentis, suggesting that humans, household pets, farm animals, wildlife and migratory birds may be the potential sources of the parasites. The findings are of use for implementing preventive measures to reduce the transmission of cryptosporidiosis to both humans and animals.

  7. Photometer for tracking a moving light source

    NASA Technical Reports Server (NTRS)

    Strawa, Anthony W. (Inventor)

    2009-01-01

    A photometer that tracks a path of a moving light source with little or no motion of the photometer components. The system includes a non-moving, truncated paraboloid of revolution, having a paraboloid axis, a paraboloid axis, a small entrance aperture, a larger exit aperture and a light-reflecting inner surface, that receives and reflects light in a direction substantially parallel to the paraboloid axis. The system also includes a light processing filter to receive and process the redirected light, and to issue the processed, redirected light as processed light, and an array of light receiving elements, at least one of which receives and measures an associated intensity of a portion of the processed light. The system tracks a light source moving along a path and produces a corresponding curvilinear image of the light source path on the array of light receiving elements. Undesired light wavelengths from the light source may be removed by coating a selected portion of the reflecting inner surface or another light receiving surface with a coating that absorbs incident light in the undesired wavelength range.

  8. Cryptosporidium sp. infections in green turtles, Chelonia mydas, as a potential source of marine waterborne oocysts in the Hawaiian Islands

    USGS Publications Warehouse

    Graczyk, T.K.; Balazs, G.H.; Work, T.M.; Aguirre, A.A.; Ellis, D.M.; Murakawa, Shawn K. K.; Morris, Robert

    1997-01-01

    For the first time, Cryptosporidium sp. oocysts were identified in fecal and intestinal samples from free-ranging marine turtles, Chelonia mydas, from the Hawaiian Islands. The oocysts produced positive reactions with commercial test kits recommended for the detection of human-infectious waterborne oocysts of Cryptosporidium parvum.

  9. Cryptosporidium: A Guide to Water Filters

    MedlinePlus

    ... Cryptosporidium Tracking in the United States CryptoNet Healthy Water Sites Healthy Water Drinking Water Healthy Swimming Global ... Button What's this? Parasites Home A Guide to Water Filters Recommend on Facebook Tweet Share Compartir Filtering ...

  10. FECAL POLLUTION, PUBLIC HEALTH AND MICROBIAL SOURCE TRACKING

    EPA Science Inventory

    Microbial source tracking (MST) seeks to provide information about sources of fecal water contamination. Without knowledge of sources, it is difficult to accurately model risk assessments, choose effective remediation strategies, or bring chronically polluted waters into complian...

  11. WATER FROM DIFFERENT SOURCES USED FOR THE IRRIGATION OF VEGETABLES TO BE MARKETED: RESEARCH ON Cryptosporidium spp., Giardia spp., AND COLIFORMS IN PARANA, BRAZIL.

    PubMed

    Tiyo, Rogerio; de Souza, Carla Zangari; Nishi, Letícia; Brustolin, Camila Fernanda; Ratti, Bianca Altrão; Falavigna Guilherme, Ana Lucia

    2015-01-01

    The aim of this work was to compare, from a parasitological ( Cryptosporidium spp. and Giardia duodenalis), bacteriological (total and thermotolerants coliforms) and physicochemical perspective, water sources used for drinking and irrigation of vegetables intended to be sold for human consumption. From January 2010 to May 2011, samples of different water sources from vegetable producing properties were collected; 100 liters for parasitological analysis, 200 mL for bacteriological analysis, and five liters for physicochemical analysis. Water samples were filtered under vacuum with a kit containing a cellulose acetate membrane filter, 1.2 µm (Millipore(r), Barueri, SP, Brazil). The material retained on the membrane was mechanically extracted and analyzed by direct immunofluorescence (Merifluor(r)kit). From 20 rural properties investigated, 10 had artesian wells (40 samples), 10 had common wells (40 samples), and one had a mine (four samples), the latter contaminated by Cryptosporidium spp. In samples from artesian wells, 90 to 130 meters depth, 42.5% were positive for total coliforms and 5.0% were identified to have abnormal coloration. From the samples of common wells, 14 to 37 meters depth, 87.5% were contaminated with total coliforms, 82.5% were positive for thermotolerant coliforms, and 12.5% had color abnormalities. We did not detect the presence of Giardia spp. or Cryptosporidium spp. in artesian and common wells. The use of artesian or common wells is an important step in the control of the spreading of zoonoses, particularly Cryptosporidium spp. and Giardia spp., as well as artesian wells for coliform control in local production of vegetables to be marketed.

  12. Tracking earthquake source evolution in 3-D

    NASA Astrophysics Data System (ADS)

    Kennett, B. L. N.; Gorbatov, A.; Spiliopoulos, S.

    2014-08-01

    Starting from the hypocentre, the point of initiation of seismic energy, we seek to estimate the subsequent trajectory of the points of emission of high-frequency energy in 3-D, which we term the `evocentres'. We track these evocentres as a function of time by energy stacking for putative points on a 3-D grid around the hypocentre that is expanded as time progresses, selecting the location of maximum energy release as a function of time. The spatial resolution in the neighbourhood of a target point can be simply estimated by spatial mapping using the properties of isochrons from the stations. The mapping of a seismogram segment to space is by inverse slowness, and thus more distant stations have a broader spatial contribution. As in hypocentral estimation, the inclusion of a wide azimuthal distribution of stations significantly enhances 3-D capability. We illustrate this approach to tracking source evolution in 3-D by considering two major earthquakes, the 2007 Mw 8.1 Solomons islands event that ruptured across a plate boundary and the 2013 Mw 8.3 event 610 km beneath the Sea of Okhotsk. In each case we are able to provide estimates of the evolution of high-frequency energy that tally well with alternative schemes, but also to provide information on the 3-D characteristics that is not available from backprojection from distant networks. We are able to demonstrate that the major characteristics of event rupture can be captured using just a few azimuthally distributed stations, which opens the opportunity for the approach to be used in a rapid mode immediately after a major event to provide guidance for, for example tsunami warning for megathrust events.

  13. Microbial Source Tracking in Adjacent Karst Springs.

    PubMed

    Ohad, Shoshanit; Vaizel-Ohayon, Dalit; Rom, Meir; Guttman, Joseph; Berger, Diego; Kravitz, Valeria; Pilo, Shlomo; Huberman, Zohar; Kashi, Yechezkel; Rorman, Efrat

    2015-08-01

    Modern man-made environments, including urban, agricultural, and industrial environments, have complex ecological interactions among themselves and with the natural surroundings. Microbial source tracking (MST) offers advanced tools to resolve the host source of fecal contamination beyond indicator monitoring. This study was intended to assess karst spring susceptibilities to different fecal sources using MST quantitative PCR (qPCR) assays targeting human, bovine, and swine markers. It involved a dual-time monitoring frame: (i) monthly throughout the calendar year and (ii) daily during a rainfall event. Data integration was taken from both monthly and daily MST profile monitoring and improved identification of spring susceptibility to host fecal contamination; three springs located in close geographic proximity revealed different MST profiles. The Giach spring showed moderate fluctuations of MST marker quantities amid wet and dry samplings, while the Zuf spring had the highest rise of the GenBac3 marker during the wet event, which was mirrored in other markers as well. The revelation of human fecal contamination during the dry season not connected to incidents of raining leachates suggests a continuous and direct exposure to septic systems. Pigpens were identified in the watersheds of Zuf, Shefa, and Giach springs and on the border of the Gaaton spring watershed. Their impact was correlated with partial detection of the Pig-2-Bac marker in Gaaton spring, which was lower than detection levels in all three of the other springs. Ruminant and swine markers were detected intermittently, and their contamination potential during the wet samplings was exposed. These results emphasized the importance of sampling design to utilize the MST approach to delineate subtleties of fecal contamination in the environment.

  14. A study of two U.S. Army installation drinking water sources and treatment systems for the removal of Giardia and Cryptosporidium

    SciTech Connect

    Kelley, M.B.; Brokaw, J.K.; Brokaw, J.K.; Warrier, P.K.

    1996-11-01

    This paper provides the results of a study of two U.S. Army installation drinking water sources and treatment systems for the removal of Giardia and Cryptosporidium. Sampling was conducted monthly for one year commencing in late March 1994 and concluding at the end of February 1995. Results of this detailed study include examination of turbidity, particle counts, and total and fecal coliforms as well as the enumeration of Giardia cysts and Cryptosporidium oocysts. Our goal, in addition to determining the levels of these pathogens in both raw and product waters, was to determine if typical operating parameters would be helpful in identifying either elevated raw water protozoa or breakthrough of either pathogen in the product water from the treatment facilities. A data summary for the results of the protozoa enumeration is a Table 1. Our results indicate frequent contamination of the raw waters at both sites by either or both pathogens. Further, we observed sporadic breakthrough of low levels of Cryptosporidium in the filtered waters of both sites. The method employed to concentrate, purify and enumerate the pathogenic protozoa is also discussed and comparisons are made to the proposed Information Collection Rule (ICR) method for detection of these microorganisms.

  15. Analysis of Solar Two Heliostat Tracking Error Sources

    SciTech Connect

    Jones, S.A.; Stone, K.W.

    1999-01-28

    This paper explores the geometrical errors that reduce heliostat tracking accuracy at Solar Two. The basic heliostat control architecture is described. Then, the three dominant error sources are described and their effect on heliostat tracking is visually illustrated. The strategy currently used to minimize, but not truly correct, these error sources is also shown. Finally, a novel approach to minimizing error is presented.

  16. Microbial source tracking: a forensic technique for microbial source identification?

    PubMed

    Stapleton, Carl M; Wyer, Mark D; Kay, David; Crowther, John; McDonald, Adrian T; Walters, Martin; Gawler, Andrew; Hindle, Terry

    2007-05-01

    As the requirements of the Water Framework Directive (WFD) and the US Clean Water Act (USCWA) for the maintenance of microbiological water quality in 'protected areas' highlight, there is a growing recognition that integrated management of point and diffuse sources of microbial pollution is essential. New information on catchment microbial dynamics and, in particular, the sources of faecal indicator bacteria found in bathing and shellfish harvesting waters is a pre-requisite for the design of any 'programme of measures' at the drainage basin scale to secure and maintain compliance with existing and new health-based microbiological standards. This paper reports on a catchment-scale microbial source tracking (MST) study in the Leven Estuary drainage basin, northwest England, an area for which quantitative faecal indicator source apportionment empirical data and land use information were also collected. Since previous MST studies have been based on laboratory trials using 'manufactured' samples or analyses of spot environmental samples without the contextual microbial flux data (under high and low flow conditions) and source information, such background data are needed to evaluate the utility of MST in USCWA total maximum daily load (TMDL) assessments or WFD 'Programmes of Measures'. Thus, the operational utility of MST remains in some doubt. The results of this investigation, using genotyping of Bacteroidetes using polymerase chain reaction (PCR) and male-specific ribonucleic acid coliphage (F + RNA coliphage) using hybridisation, suggest some discrimination is possible between livestock- and human-derived faecal indicator concentrations but, in inter-grade areas, the degree to which the tracer picture reflected the land use pattern and probable faecal indicator loading were less distinct. Interestingly, the MST data was more reliable on high flow samples when much of the faecal indicator flux from catchment systems occurs. Whilst a useful supplementary tool, the MST

  17. Comparison of diagnostic techniques for the detection of Cryptosporidium oocysts in animal samples.

    PubMed

    Ezzaty Mirhashemi, Marzieh; Zintl, Annetta; Grant, Tim; Lucy, Frances E; Mulcahy, Grace; De Waal, Theo

    2015-01-01

    While a large number of laboratory methods for the detection of Cryptosporidium oocysts in faecal samples are now available, their efficacy for identifying asymptomatic cases of cryptosporidiosis is poorly understood. This study was carried out to determine a reliable screening test for epidemiological studies in livestock. In addition, three molecular tests were compared to identify Cryptosporidium species responsible for the infection in cattle, sheep and horses. A variety of diagnostic tests including microscopic (Kinyoun's staining), immunological (Direct Fluorescence Antibody tests or DFAT), enzyme-linked immunosorbent assay (ELISA), and molecular methods (nested PCR) were compared to assess their ability to detect Cryptosporidium in cattle, horse and sheep faecal samples. The results indicate that the sensitivity and specificity of each test is highly dependent on the input samples; while Kinyoun's and DFAT proved to be reliable screening tools for cattle samples, DFAT and PCR analysis (targeted at the 18S rRNA gene fragment) were more sensitive for screening sheep and horse samples. Finally different PCR primer sets targetedat the same region resulted in the preferential amplification of certain Cryptosporidium species when multiple species were present in the sample. Therefore, for identification of Cryptosporidium spp. in the event of asymptomatic cryptosporidiosis, the combination of different 18S rRNA nested PCR primer sets is recommended for further epidemiological applications and also tracking the sources of infection.

  18. Comparison of diagnostic techniques for the detection of Cryptosporidium oocysts in animal samples

    PubMed Central

    Mirhashemi, Marzieh Ezzaty; Zintl, Annetta; Grant, Tim; Lucy, Frances E.; Mulcahy, Grace; De Waal, Theo

    2015-01-01

    While a large number of laboratory methods for the detection of Cryptosporidium oocysts in faecal samples are now available, their efficacy for identifying asymptomatic cases of cryptosporidiosis is poorly understood. This study was carried out to determine a reliable screening test for epidemiological studies in livestock. In addition, three molecular tests were compared to identify Cryptosporidium species responsible for the infection in cattle, sheep and horses. A variety of diagnostic tests including microscopic (Kinyoun's staining), immunological (Direct Fluorescence Antibody tests or DFAT), enzyme-linked immunosorbent assay (ELISA), and molecular methods (nested PCR) were compared to assess their ability to detect Cryptosporidium in cattle, horse and sheep faecal samples. The results indicate that the sensitivity and specificity of each test is highly dependent on the input samples; while Kinyoun's and DFAT proved to be reliable screening tools for cattle samples, DFAT and PCR analysis (targeted at the 18S rRNA gene fragment) were more sensitive for screening sheep and horse samples. Finally different PCR primer sets targeted at the same region resulted in the preferential amplification of certain Cryptosporidium species when multiple species were present in the sample. Therefore, for identification of Cryptosporidium spp. in the event of asymptomatic cryptosporidiosis, the combination of different 18S rRNA nested PCR primer sets is recommended for further epidemiological applications and also tracking the sources of infection. PMID:25662435

  19. Tracking Moving Acoustic Sources With a Network of Sensors

    DTIC Science & Technology

    2002-10-01

    Tracking Moving Acoustic Sources With a Network of Sensors by Richard J. Kozick and Brian M. Sadler ARL-TR-2750 October 2002 Approved for public...October 2002 Tracking Moving Acoustic Sources With a Network of Sensors Richard J. Kozick Bucknell University, Electrical Engineering Department Brian M...Model for a Nonmoving Source 4 2.1 Cramér-Rao Bound (CRB) . . . . . . . . . . . . . . . . . . . . 6 2.2 Examples

  20. Cryptosporidium parvum and Cyclospora cayetanensis: a review of laboratory methods for detection of these waterborne parasites.

    PubMed

    Quintero-Betancourt, Walter; Peele, Emily R; Rose, Joan B

    2002-05-01

    Cryptosporidium and Cyclospora are obligate, intracellular, coccidian protozoan parasites that infest the gastrointestinal tract of humans and animals causing severe diarrhea illness. In this paper, we present an overview of the conventional and more novel techniques that are currently available to detect Cryptosporidium and Cyclospora in water. Conventional techniques and new immunological and genetic/molecular methods make it possible to assess the occurrence, prevalence, virulence (to a lesser extent), viability, levels, and sources of waterborne protozoa. Concentration, purification, and detection are the three key steps in all methods that have been approved for routine monitoring of waterborne oocysts. These steps have been optimized to such an extent that low levels of naturally occurring Cryptosporidium oocysts can be efficiently recovered from water. The filtration systems developed in the US and Europe trap oocysts more effectively and are part of the standard methodologies for environmental monitoring of Cryptosporidium oocysts in source and treated water. Purification techniques such as immunomagnetic separation and flow cytometry with fluorescent activated cell sorting impart high capture efficiency and selective separation of oocysts from sample debris. Monoclonal antibodies with higher avidity and specificity to oocysts in water concentrates have significantly improved the detection and enumeration steps. To date, PCR-based detection methods allow us to differentiate the human pathogenic Cryptosporidium parasites from those that do not infect humans, and to track the source of oocyst contamination in the environment. Cell culture techniques are now used to examine oocyst viability. While fewer studies have focused on Cyclospora cayetanensis, the parasite has been successfully detected in drinking water and wastewater using current methods to recover Cryptosporidium oocysts. More research is needed for monitoring of Cyclospora in the environment

  1. Multi-source energy harvester for wildlife tracking

    NASA Astrophysics Data System (ADS)

    Wu, You; Zuo, Lei; Zhou, Wanlu; Liang, Changwei; McCabe, Michael

    2014-03-01

    Sufficient power supply to run GPS machinery and transmit data on a long-term basis remains to be the key challenge for wildlife tracking technology. Traditional way of replacing battery periodically is not only time and money consuming but also dangerous to live-trapping wild animals. In this paper, an innovative wildlife tracking device with multi-source energy harvester with advantage of high efficiency and reliability is investigated and developed. This multi-source energy harvester entails a solar energy harvester and an innovative rotational electromagnetic energy harvester is mounted on the "wildlife tracking collar" which will remarkably extend the duration of wild life tracking. A feedforward and feedback control of DC-DC converter circuit is adopted to passively realize the Maximum Power Point Tracking (MPPT) logic for the solar energy harvester. The rotational electromagnetic energy harvester can mechanically rectify the irregular bidirectional motion into unidirectional motion has been modeled and demonstrated.

  2. Sediment Source Tracking in the Georgia Piedmont

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Preliminary results on the source of suspended sediment in a southern Piedmont stream are presented. Nuclear fallout radionuclide 137Cs and three other natural tracers were used to estimate the relative contribution of bank and upland sediment sources. Tracer concentrations were determined in potent...

  3. THE EPA MICROBIAL SOURCE TRACKING DOCUMENT

    EPA Science Inventory

    Beach closures or violations of total maximum daily loads of fecal organisms in watersheds frequently generate a need to identify the major sources of contamination or, at least, determine whether the source is human or animal. A few years ago E. coli ribotyping was the only met...

  4. Use of Biological and Non-biological Surrogates for Evaluating Cryptosporidium Removal by Filtration

    EPA Science Inventory

    Water treatment plants are currently facing increasing challenges in monitoring Cryptosporidium in source and treated water because of complex analytical techniques and associated health risks. Surrogates may be easier to analyze than Cryptosporidium, but they must also be reliab...

  5. Geoacoustic and source tracking using particle filtering: experimental results.

    PubMed

    Yardim, Caglar; Gerstoft, Peter; Hodgkiss, William S

    2010-07-01

    A particle filtering (PF) approach is presented for performing sequential geoacoustic inversion of a complex ocean acoustic environment using a moving acoustic source. This approach treats both the environmental parameters [e.g., water column sound speed profile (SSP), water depth, sediment and bottom parameters] at the source location and the source parameters (e.g., source depth, range and speed) as unknown random variables that evolve as the source moves. This allows real-time updating of the environment and accurate tracking of the moving source. As a sequential Monte Carlo technique that operates on nonlinear systems with non-Gaussian probability densities, the PF is an ideal algorithm to perform tracking of environmental and source parameters, and their uncertainties via the evolving posterior probability densities. The approach is demonstrated on both simulated data in a shallow water environment with a sloping bottom and experimental data collected during the SWellEx-96 experiment.

  6. Cryptosporidium and Giardia removal by secondary and tertiary wastewater treatment.

    PubMed

    Taran-Benshoshan, Marina; Ofer, Naomi; Dalit, Vaizel-Ohayon; Aharoni, Avi; Revhun, Menahem; Nitzan, Yeshayahu; Nasser, Abidelfatah M

    2015-01-01

    Wastewater disposal may be a source of environmental contamination by Cryptosporidium and Giardia. This study was conducted to evaluate the prevalence of Cryptosporidium oocysts and Giardia cysts in raw and treated wastewater effluents. A prevalence of 100% was demonstrated for Giardia cysts in raw wastewater, at a concentration range of 10 to 12,225 cysts L(-1), whereas the concentration of Cryptosporidium oocysts in raw wastewater was 4 to 125 oocysts L(-1). The removal of Giardia cysts by secondary and tertiary treatment processes was greater than those observed for Cryptosporidium oocysts and turbidity. Cryptosporidium and Giardia were present in 68.5% and 76% of the tertiary effluent samples, respectively, at an average concentration of 0.93 cysts L(-1) and 9.94 oocysts L(-1). A higher detection limit of Cryptosporidium oocysts in wastewater was observed for nested PCR as compared to immune fluorescent assay (IFA). C. hominis was found to be the dominant genotype in wastewater effluents followed by C. parvum and C. andersoni or C. muris. Giardia was more prevalent than Cryptosporidium in the studied community and treatment processes were more efficient for the removal of Giardia than Cryptosporidium. Zoonotic genotypes of Cryptosporidium were also present in the human community. To assess the public health significance of Cryptosporidium oocysts present in tertiary effluent, viability (infectivity) needs to be assessed.

  7. Tracking the sources of tropospheric ozone

    NASA Astrophysics Data System (ADS)

    Butler, T. M.; Churkina, G.; Coates, J.; Grote, R.; Mar, K.; von Schneidemesser, E.; Zhu, S.

    2013-12-01

    Tropospheric ozone is a harmful pollutant with adverse effects on human health and ecosystems. As well as these effects, tropospheric ozone is also a powerful greenhouse gas, with an anthropogenic radiative forcing one quarter of that of CO2. Along with methane and atmospheric aerosol, tropospheric ozone belongs to the so-called Short Lived Climate forcing Pollutants, or SLCP. Recent work has shown that efforts to reduce concentrations of SLCP in the atmosphere have the potential to slow the rate of near-term climate change, while simultaneously improving public health and reducing crop losses. Unlike many other SLCP, tropospehric ozone is not directly emitted, but is instead influenced by two distinct sources: transport of air from the ozone-rich stratosphere; and photochemical production in the troposphere from the emitted precursors NOx (oxides of nitrogen), CO (Carbon Monoxide), and VOC (volatile organic compounds, including methane). Better understanding of the relationship between ozone production and the emissions of its precursors is essential for the development of targeted emission reduction strategies. Several modeling methods have been employed to relate the production of tropospheric ozone to emissions of its precursors; emissions perturbation, tagging, and adjoint sensitivity methods all deliver complementary information about modelled ozone production. Most studies using tagging methods have focused on attribution of tropospheric ozone production to emissions of NOx, even though perturbation methods have suggested that tropospheric ozone is also sensitive to VOC, particularly methane. In this set of studies we examine the attribution of tropospheric ozone to emissions of VOC using a tagging approach, whereby each VOC oxidation intermediate in model chemical mechanisms is tagged with the identity of its primary emitted compound, allowing modelled ozone production to be directly attributed to all emitted VOCs in the model. Using a global model we

  8. 10 CFR 32.201 - Serialization of nationally tracked sources.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 1 2010-01-01 2010-01-01 false Serialization of nationally tracked sources. 32.201 Section 32.201 Energy NUCLEAR REGULATORY COMMISSION SPECIFIC DOMESTIC LICENSES TO MANUFACTURE OR TRANSFER CERTAIN ITEMS CONTAINING BYPRODUCT MATERIAL Specifically Licensed Items § 32.201 Serialization...

  9. Coherence among Different Microbial Source Tracking Markers in a Small Agricultural Stream with or without Livestock Exclusion Practices

    PubMed Central

    Wilkes, Graham; Brassard, Julie; Edge, Thomas A.; Gannon, Victor; Jokinen, Cassandra C.; Jones, Tineke H.; Marti, Romain; Neumann, Norman F.; Ruecker, Norma J.; Sunohara, Mark; Topp, Edward

    2013-01-01

    Over 1,400 water samples were collected biweekly over 6 years from an intermittent stream protected and unprotected from pasturing cattle. The samples were monitored for host-specific Bacteroidales markers, Cryptosporidium species/genotypes, viruses and coliphages associated with humans or animals, and bacterial zoonotic pathogens. Ruminant Bacteroidales markers did not increase within the restricted cattle access reach of the stream, whereas the ruminant Bacteroidales marker increased significantly in the unrestricted cattle access reach. Human Bacteroidales markers significantly increased downstream of homes where septic issues were documented. Wildlife Bacteroidales markers were detected downstream of the cattle exclusion practice where stream and riparian habitat was protected, but detections decreased after the unrestricted pasture, where the stream and riparian zone was unprotected from livestock. Detection of a large number of human viruses was shown to increase downstream of homes, and similar trends were observed for the human Bacteroidales marker. There was considerable interplay among biomarkers with stream flow, season, and the cattle exclusion practices. There were no to very weak associations with Bacteroidales markers and bacterial, viral, and parasitic pathogens. Overall, discrete sample-by-sample coherence among the different microbial source tracking markers that expressed a similar microbial source was minimal, but spatial trends were physically meaningful in terms of land use (e.g., beneficial management practice) effects on sources of fecal pollution. PMID:23913430

  10. Coherence among different microbial source tracking markers in a small agricultural stream with or without livestock exclusion practices.

    PubMed

    Wilkes, Graham; Brassard, Julie; Edge, Thomas A; Gannon, Victor; Jokinen, Cassandra C; Jones, Tineke H; Marti, Romain; Neumann, Norman F; Ruecker, Norma J; Sunohara, Mark; Topp, Edward; Lapen, David R

    2013-10-01

    Over 1,400 water samples were collected biweekly over 6 years from an intermittent stream protected and unprotected from pasturing cattle. The samples were monitored for host-specific Bacteroidales markers, Cryptosporidium species/genotypes, viruses and coliphages associated with humans or animals, and bacterial zoonotic pathogens. Ruminant Bacteroidales markers did not increase within the restricted cattle access reach of the stream, whereas the ruminant Bacteroidales marker increased significantly in the unrestricted cattle access reach. Human Bacteroidales markers significantly increased downstream of homes where septic issues were documented. Wildlife Bacteroidales markers were detected downstream of the cattle exclusion practice where stream and riparian habitat was protected, but detections decreased after the unrestricted pasture, where the stream and riparian zone was unprotected from livestock. Detection of a large number of human viruses was shown to increase downstream of homes, and similar trends were observed for the human Bacteroidales marker. There was considerable interplay among biomarkers with stream flow, season, and the cattle exclusion practices. There were no to very weak associations with Bacteroidales markers and bacterial, viral, and parasitic pathogens. Overall, discrete sample-by-sample coherence among the different microbial source tracking markers that expressed a similar microbial source was minimal, but spatial trends were physically meaningful in terms of land use (e.g., beneficial management practice) effects on sources of fecal pollution.

  11. Fecal Sterol and Runoff Analysis for Nonpoint Source Tracking.

    PubMed

    Fahrenfeld, N L; Del Monaco, N; Coates, J T; Elzerman, A W

    2016-01-01

    Fecal pollution source identification is needed to quantify risk, target installation of source controls, and assess performance of best management practices in impaired surface waters. Sterol analysis is a chemical method for fecal source tracking that allows for differentiation between several fecal pollution sources. The objectives of this study were to use these chemical tracers for quantifying human fecal inputs in a mixed-land-use watershed without point sources of pollution and to determine the relationship between land use and sterol ratios. Fecal sterol analysis was performed on bed and suspended sediment from impaired streams. Human fecal signatures were found at sites with sewer overflow and septic inputs. Different sterol ratios used to indicate human fecal pollution varied in their sensitivity. Next, geospatial data was used to determine the runoff volumes associated with each land-use category in the watersheds. Fecal sterol ratios were compared between sampling locations and correlations were tested between ratio values and percentage of runoff for a given land-use category. Correlation was not observed between percentage of runoff from developed land and any of the five tested human-indicating sterol ratios in streambed sediments, confirming that human fecal inputs were not evenly distributed across the urban landscape. Several practical considerations for adopting this chemical method for microbial source tracking in small watersheds are discussed. Results indicate that sterol analysis is useful for identifying the location of human fecal nonpoint-source inputs.

  12. Molecular-Based Detection Systems for Cryptosporidium Oocysts

    EPA Science Inventory

    The presentation describes on-going studies in collaboration with US EPA Region 2, 3, and the CDC on identifying sources of Cryptosporidium oocyst contamination in source waters using conventional and real-time PCR approaches.

  13. Multiple approaches to microbial source tracking in tropical northern Australia

    PubMed Central

    Neave, Matthew; Luter, Heidi; Padovan, Anna; Townsend, Simon; Schobben, Xavier; Gibb, Karen

    2014-01-01

    Microbial source tracking is an area of research in which multiple approaches are used to identify the sources of elevated bacterial concentrations in recreational lakes and beaches. At our study location in Darwin, northern Australia, water quality in the harbor is generally good, however dry-season beach closures due to elevated Escherichia coli and enterococci counts are a cause for concern. The sources of these high bacteria counts are currently unknown. To address this, we sampled sewage outfalls, other potential inputs, such as urban rivers and drains, and surrounding beaches, and used genetic fingerprints from E. coli and enterococci communities, fecal markers and 454 pyrosequencing to track contamination sources. A sewage effluent outfall (Larrakeyah discharge) was a source of bacteria, including fecal bacteria that impacted nearby beaches. Two other treated effluent discharges did not appear to influence sites other than those directly adjacent. Several beaches contained fecal indicator bacteria that likely originated from urban rivers and creeks within the catchment. Generally, connectivity between the sites was observed within distinct geographical locations and it appeared that most of the bacterial contamination on Darwin beaches was confined to local sources. PMID:25224738

  14. Multiple approaches to microbial source tracking in tropical northern Australia.

    PubMed

    Neave, Matthew; Luter, Heidi; Padovan, Anna; Townsend, Simon; Schobben, Xavier; Gibb, Karen

    2014-12-01

    Microbial source tracking is an area of research in which multiple approaches are used to identify the sources of elevated bacterial concentrations in recreational lakes and beaches. At our study location in Darwin, northern Australia, water quality in the harbor is generally good, however dry-season beach closures due to elevated Escherichia coli and enterococci counts are a cause for concern. The sources of these high bacteria counts are currently unknown. To address this, we sampled sewage outfalls, other potential inputs, such as urban rivers and drains, and surrounding beaches, and used genetic fingerprints from E. coli and enterococci communities, fecal markers and 454 pyrosequencing to track contamination sources. A sewage effluent outfall (Larrakeyah discharge) was a source of bacteria, including fecal bacteria that impacted nearby beaches. Two other treated effluent discharges did not appear to influence sites other than those directly adjacent. Several beaches contained fecal indicator bacteria that likely originated from urban rivers and creeks within the catchment. Generally, connectivity between the sites was observed within distinct geographical locations and it appeared that most of the bacterial contamination on Darwin beaches was confined to local sources.

  15. Molecular characterization of Cryptosporidium and Giardia occurring in natural water bodies in Poland.

    PubMed

    Adamska, Małgorzata

    2015-02-01

    Cryptosporidium and Giardia protozoa are zoonotic parasites that cause human gastroenteritis and can be transmitted to human through the fecal-oral route and water or food. Several species belong to these genera and their resistant forms occur in water, but only some of them are infectious to human. Health risk depends on the occurrence of infectious Cryptosporidium and Giardia species and genotypes in water, and only molecular techniques allow detecting them, as well as enable to identify the contamination source. In this work, genotyping and phylogenetic analysis have been performed on the basis of 18S rDNA and ß-giardin genes sequences of Cryptosporidium and Giardia, respectively, in order to provide the molecular characterization of these parasites detected earlier in five natural water bodies in Poland and to track possible sources of their (oo)cysts in water. Genotyping revealed a high similarity (over 99 up to 100 %) of analyzed sequences to cattle genotype of C. parvum isolated from cattle and human and to G. intestinalis assemblage B isolated from human. The sequences obtained by others originated from patients with clinical symptoms of cryptosporidiosis or giardiasis and/or with the infection confirmed by different methods. The contamination of three examined lakes is probably human-originated, while the sources of contamination of two remaining lakes are wild and domestic animals. Obtained phylogenetic trees support suggestions of other authors that the bovine genotype of C. parvum should be a separate species, as well as A and B assemblages of G. intestinalis.

  16. An open source mobile platform for psychophysiological self tracking.

    PubMed

    Gaggioli, Andrea; Cipresso, Pietro; Serino, Silvia; Pioggia, Giovanni; Tartarisco, Gennaro; Baldus, Giovanni; Corda, Daniele; Riva, Giuseppe

    2012-01-01

    Self tracking is a recent trend in e-health that refers to the collection, elaboration and visualization of personal health data through ubiquitous computing tools such as mobile devices and wearable sensors. Here, we describe the design of a mobile self-tracking platform that has been specifically designed for clinical and research applications in the field of mental health. The smartphone-based application allows collecting a) self-reported feelings and activities from pre-programmed questionnaires; b) electrocardiographic (ECG) data from a wireless sensor platform worn by the user; c) movement activity information obtained from a tri-axis accelerometer embedded in the wearable platform. Physiological signals are further processed by the application and stored on the smartphone's memory. The mobile data collection platform is free and released under an open source licence to allow wider adoption by the research community (download at: http://sourceforge.net/projects/psychlog/).

  17. Source Tracking of Nitrous Oxide using A Quantum Cascade ...

    EPA Pesticide Factsheets

    Nitrous oxide is an important greenhouse gas and ozone depleting substance. Nitrification and denitrification are two major biological pathways that are responsible for soil emissions of N2O. However, source tracking of in-situ or laboratory N2O production is still challenging to soil scientists. The objective of this study was to introduce the use of a new technology, quantum cascade laser (QCL) spectroscopy, which allows for significantly improved accuracy and precision to continuously measure real-time N2O for source tracking. This data provides important emission inventory information to air quality and atmospheric chemistry models. The task demonstrated that QCL spectroscopy can measure the flux of nitrous oxide at ambient and well as elevated concentrations in real time. The fractionation of the nitrous oxide produced by microbial processing of nitrate can be measured and characterized as isotopic signatures related to the nitrifying or denitrifying state of the microbial communities. This has important implications for monitoring trace gases in the atmosphere. The data produced by this system will provide clients including the air quality and climate change communities with needed information on the sources and strengths of N2O emissions for modeling and research into mitigation strategies to reduce overall GHG emissions in agricultural systems.

  18. Molecular characterization of bacteriophages for microbial source tracking in Korea.

    PubMed

    Lee, Jung Eun; Lim, Mi Young; Kim, Sei Yoon; Lee, Sunghee; Lee, Heetae; Oh, Hyun-Myung; Hur, Hor-Gil; Ko, Gwangpyo

    2009-11-01

    We investigated coliphages from various fecal sources, including humans and animals, for microbial source tracking in South Korea. Both somatic and F+-specific coliphages were isolated from 43 fecal samples from farms, wild animal habitats, and human wastewater plants. Somatic coliphages were more prevalent and abundant than F+ coliphages in all of the tested fecal samples. We further characterized 311 F+ coliphage isolates using RNase sensitivity assays, PCR and reverse transcription-PCR, and nucleic acid sequencing. Phylogenetic analyses were performed based on the partial nucleic acid sequences of 311 F+ coliphages from various sources. F+ RNA coliphages were most prevalent among geese (95%) and were least prevalent in cows (5%). Among the genogroups of F+ RNA coliphages, most F+ coliphages isolated from animal fecal sources belonged to either group I or group IV, and most from human wastewater sources were in group II or III. Some of the group I coliphages were present in both human and animal source samples. F+ RNA coliphages isolated from various sources were divided into two main clusters. All F+ RNA coliphages isolated from human wastewater were grouped with Qbeta-like phages, while phages isolated from most animal sources were grouped with MS2-like phages. UniFrac significance statistical analyses revealed significant differences between human and animal bacteriophages. In the principal coordinate analysis (PCoA), F+ RNA coliphages isolated from human waste were distinctively separate from those isolated from other animal sources. However, F+ DNA coliphages were not significantly different or separate in the PCoA. These results demonstrate that proper analysis of F+ RNA coliphages can effectively distinguish fecal sources.

  19. PAIRED CITY CRYPTOSPORIDIUM SEROSURVEY

    EPA Science Inventory

    In 1996, serological responses to two Cryptosporidium antigens were determined for 200 Las Vegas (LV), Nevada, and 200 Albuquerque, New Mexico, blood donors to evaluate associations between endemic infections, water exposures, and other risk factors. LV uses chlorinated filtered...

  20. METHODS FOR DETECTION OF CRYPTOSPORIDIUM SP. AND GIARDIA SP.

    EPA Science Inventory

    There have been several waterborne outbreaks of giardiasis caused by infection with Giardia lamblia, and cryptosporidiosis, caused by infection with Cryptosporidium parvum. These outbreaks have created a need to detect these organisms in source and finished drinking water. The pr...

  1. Cryptosporidium Pathogenicity and Virulence

    PubMed Central

    Bouzid, Maha; Chalmers, Rachel M.; Tyler, Kevin M.

    2013-01-01

    Cryptosporidium is a protozoan parasite of medical and veterinary importance that causes gastroenteritis in a variety of vertebrate hosts. Several studies have reported different degrees of pathogenicity and virulence among Cryptosporidium species and isolates of the same species as well as evidence of variation in host susceptibility to infection. The identification and validation of Cryptosporidium virulence factors have been hindered by the renowned difficulties pertaining to the in vitro culture and genetic manipulation of this parasite. Nevertheless, substantial progress has been made in identifying putative virulence factors for Cryptosporidium. This progress has been accelerated since the publication of the Cryptosporidium parvum and C. hominis genomes, with the characterization of over 25 putative virulence factors identified by using a variety of immunological and molecular techniques and which are proposed to be involved in aspects of host-pathogen interactions from adhesion and locomotion to invasion and proliferation. Progress has also been made in the contribution of host factors that are associated with variations in both the severity and risk of infection. Here we provide a review comprised of the current state of knowledge on Cryptosporidium infectivity, pathogenesis, and transmissibility in light of our contemporary understanding of microbial virulence. PMID:23297262

  2. Fecal source tracking in water using a mitochondrial DNA microarray.

    PubMed

    Vuong, Nguyet-Minh; Villemur, Richard; Payment, Pierre; Brousseau, Roland; Topp, Edward; Masson, Luke

    2013-01-01

    A mitochondrial-based microarray (mitoArray) was developed for rapid identification of the presence of 28 animals and one family (cervidae) potentially implicated in fecal pollution in mixed activity watersheds. Oligonucleotide probes for genus or subfamily-level identification were targeted within the 12S rRNA - Val tRNA - 16S rRNA region in the mitochondrial genome. This region, called MI-50, was selected based on three criteria: 1) the ability to be amplified by universal primers 2) these universal primer sequences are present in most commercial and domestic animals of interest in source tracking, and 3) that sufficient sequence variation exists within this region to meet the minimal requirements for microarray probe discrimination. To quantify the overall level of mitochondrial DNA (mtDNA) in samples, a quantitative-PCR (Q-PCR) universal primer pair was also developed. Probe validation was performed using DNA extracted from animal tissues and, for many cases, animal-specific fecal samples. To reduce the amplification of potentially interfering fish mtDNA sequences during the MI-50 enrichment step, a clamping PCR method was designed using a fish-specific peptide nucleic acid. DNA extracted from 19 water samples were subjected to both array and independent PCR analyses. Our results confirm that the mitochondrial microarray approach method could accurately detect the dominant animals present in water samples emphasizing the potential for this methodology in the parallel scanning of a large variety of animals normally monitored in fecal source tracking.

  3. [Microbial source tracking of water fecal pollution: a review].

    PubMed

    Feng, Guan-da; Deng, Ming-rong; Zhu, Hong-hui; Guo, Jun; Zhang, Xi; Zhu, Chang-xiong; Liang, Hao-liang

    2010-12-01

    Livestock feces and domestic sewage are the one of the main factors inducing water pollution, while the identification of the pollution source is particularly important in pollution control and management. Because of this, microbial source tracking (MST) has recently been paid more and more attention by the related researchers around the world. In this paper, the research progress of two types of MST methods, their advantages and disadvantages, and existing problems in application were reviewed and discussed. It was considered that in the library- and culture-dependent MST methods, PCR genotyping based on repetitive sequences was most practicable, while in the library- and culture-independent MST methods, PCR-DGGE based on the gene of specificity in Escherichia coli had a very glaring sight. Future researches should be more focused on the library- and culture-independent MST, and the combination of library- and culture-dependent MST with library- and culture-independent MST could make the tracking results more credible.

  4. Bacteriophages infecting Bacteroides as a marker for microbial source tracking.

    PubMed

    Jofre, Joan; Blanch, Anicet R; Lucena, Francisco; Muniesa, Maite

    2014-05-15

    Bacteriophages infecting certain strains of Bacteroides are amid the numerous procedures proposed for tracking the source of faecal pollution. These bacteriophages fulfil reasonably well most of the requirements identified as appropriate for a suitable marker of faecal sources. Thus, different host strains are available that detect bacteriophages preferably in water contaminated with faecal wastes corresponding to different animal species. For phages found preferably in human faecal wastes, which are the ones that have been more extensively studied, the amounts of phages found in waters contaminated with human fecal samples is reasonably high; these amounts are invariable through the time; their resistance to natural and anthropogenic stressors is comparable to that of other relatively resistant indicator of faecal pollution such us coliphages; the abundance ratios of somatic coliphages and bacteriophages infecting Bacteroides thetaiotaomicron GA17 are unvarying in recent and aged contamination; and standardised detection methods exist. These methods are easy, cost effective and provide data susceptible of numerical analysis. In contrast, there are some uncertainties regarding their geographical stability, and consequently suitable hosts need to be isolated for different geographical areas. However, a feasible method has been described to isolate suitable hosts in a given geographical area. In summary, phages infecting Bacteroides are a marker of faecal sources that in our opinion merits being included in the "toolbox" for microbial source tracking. However, further research is still needed in order to make clear some uncertainties regarding some of their characteristics and behaviour, to compare their suitability to the one of emerging methods such us targeting Bacteroidetes by qPCR assays; or settling molecular methods for their determination.

  5. Mechanical transmission of Cryptosporidium parvum oocysts by flies.

    PubMed

    Graczyk, Thaddeus K; Grimes, Barbara H; Knight, Ronald; Szostakowska, Beata; Kruminis-Lozowska, Wiesława; Racewicz, Maria; Tamang, Leena; Dasilva, Alexandre J; Myjak, Przemysław

    2004-01-01

    Long term field studies and laboratory experiments demonstrated that synanthropic filth flies can mechanically transmit infectious oocysts of Cryptosporidium parvum, an anthropozoonotic protozoan parasite which significantly contributes to the mortality of immunocompromised or immunosuppressed people. C. parvum oocysts are acquired from unhygienic sources, and can pass trough fly gastrointestinal track without alteration of their infectivity and can be subsequently deposited on visited surfaces. Transmission of the oocysts by adult flies occurs via: (1) mechanical dislodgement from the exoskeleton; (2) fecal deposition; and (3) regurgitation, i.e., vomits. Filth flies can cause human or animal cryptosporidiosis via deposition of infectious oocysts on the visited foodstuf, and the biology and ecology of synanthropic filth flies indicate that their potential for mechanical transmission of C. parvum is high.

  6. Global modelling of Cryptosporidium in surface water

    NASA Astrophysics Data System (ADS)

    Vermeulen, Lucie; Hofstra, Nynke

    2016-04-01

    Introduction Waterborne pathogens that cause diarrhoea, such as Cryptosporidium, pose a health risk all over the world. In many regions quantitative information on pathogens in surface water is unavailable. Our main objective is to model Cryptosporidium concentrations in surface waters worldwide. We present the GloWPa-Crypto model and use the model in a scenario analysis. A first exploration of global Cryptosporidium emissions to surface waters has been published by Hofstra et al. (2013). Further work has focused on modelling emissions of Cryptosporidium and Rotavirus to surface waters from human sources (Vermeulen et al 2015, Kiulia et al 2015). A global waterborne pathogen model can provide valuable insights by (1) providing quantitative information on pathogen levels in data-sparse regions, (2) identifying pathogen hotspots, (3) enabling future projections under global change scenarios and (4) supporting decision making. Material and Methods GloWPa-Crypto runs on a monthly time step and represents conditions for approximately the year 2010. The spatial resolution is a 0.5 x 0.5 degree latitude x longitude grid for the world. We use livestock maps (http://livestock.geo-wiki.org/) combined with literature estimates to calculate spatially explicit livestock Cryptosporidium emissions. For human Cryptosporidium emissions, we use UN population estimates, the WHO/UNICEF JMP sanitation country data and literature estimates of wastewater treatment. We combine our emissions model with a river routing model and data from the VIC hydrological model (http://vic.readthedocs.org/en/master/) to calculate concentrations in surface water. Cryptosporidium survival during transport depends on UV radiation and water temperature. We explore pathogen emissions and concentrations in 2050 with the new Shared Socio-economic Pathways (SSPs) 1 and 3. These scenarios describe plausible future trends in demographics, economic development and the degree of global integration. Results and

  7. Occurrence and potential health risk of Cryptosporidium and Giardia in different water catchments in Belgium.

    PubMed

    Ehsan, Amimul; Geurden, Thomas; Casaert, Stijn; Paulussen, Jef; De Coster, Lut; Schoemaker, Toon; Chalmers, Rachel; Grit, Grietje; Vercruysse, Jozef; Claerebout, Edwin

    2015-02-01

    Human wastewater and livestock can contribute to contamination of surface water with Cryptosporidium and Giardia. In countries where a substantial proportion of drinking water is produced from surface water, e.g., Belgium, this poses a constant threat on drinking water safety. Our objective was to monitor the presence of Cryptosporidium and Giardia in different water catchment sites in Belgium and to discriminate between (oo)cysts from human or animal origin using genotyping. Monthly samples were collected from raw water and purified drinking water at four catchment sites. Cryptosporidium and Giardia were detected using USEPA method 1623 and positive samples were genotyped. No contamination was found in purified water at any site. In three catchments, only low numbers of (oo)cysts were recovered from raw water samples (<1/liter), but raw water samples from one catchment site were frequently contaminated with Giardia (92 %) and Cryptosporidium (96 %), especially in winter and spring. Genotyping of Giardia in 38 water samples identified the presence of Giardia duodenalis assemblage AI, AII, BIV, BIV-like, and E. Cryptosporidium andersoni, Cryptosporidium suis, Cryptosporidium horse genotype, Cryptosporidium parvum, and Cryptosporidium hominis were detected. The genotyping results suggest that agriculture may be a more important source of surface water contamination than human waste in this catchment. In catchment sites with contaminated surface water, such as the Blankaart, continuous monitoring of treated water for the presence of Cryptosporidium and Giardia would be justified and (point) sources of surface water contamination should be identified.

  8. Using Microbial Source Tracking to Enhance Environmental Stewardship of Agriculture

    NASA Astrophysics Data System (ADS)

    Martin, Sherry; Rose, Joan; Flood, Matthew; Aw, Tiong; Hyndman, David

    2016-04-01

    Large scale agriculture relies on the application of chemical fertilizers and animal manure. It is well known that nutrients in excess of a plant's uptake and soil retention capacity can travel to nearby waterways via surface run-off and groundwater pathways, indirectly fertilizing these aquatic ecosystems. It has not yet been possible to distinguish water quality impacts of fertilizer from those derived from human and animal waste sources. However, new microbial source tracking (MST) tools allow specific identification of fecal pollution. Our objective was to examine pollution risks at the regional scale using MST, mapping and classification and regression tree analysis. We present results Bovine M2 genetic marker data from three flow regimes (baseflow, snow melt, and post-planting rain). Key landscape characteristics were related to the presence of the bovine markers and appear to be related to fate and transport. Impacts at this regional watershed scale will be discussed. Our research aims to identify the impacts of agricultural management practices on water quality by linking nutrient concentrations with fecal pollution sources. We hope that our research will provide guidance that will help improve water quality through agricultural best management practices to reduce pathogen contamination.

  9. Recommendations following a multi-laboratory comparison of microbial source tracking methods

    EPA Science Inventory

    Microbial source tracking (MST) methods are under development to provide resource managers with tools to identify sources of fecal contamination in water. Some of the most promising methods currently under development were recently evaluated in the Source Identification Protocol ...

  10. New developments in Cryptosporidium research.

    PubMed

    Ryan, Una; Hijjawi, Nawal

    2015-05-01

    Cryptosporidium is an enteric parasite that is considered the second greatest cause of diarrhoea and death in children after rotavirus. Currently, 27 species are recognised as valid and of these, Cryptosporidium hominis and Cryptosporidium parvum are responsible for the majority of infections in humans. Molecular and biological studies indicate that Cryptosporidium is more closely related to gregarine parasites rather than to coccidians. The identification of gregarine-like gamont stages and the ability of Cryptosporidium to complete its life cycle in the absence of host cells further confirm its relationship with gregarines. This opens new avenues into the investigation of pathogenesis, epidemiology, treatment and control of Cryptosporidium. Effective drug treatments and vaccines are not yet available due, in part, to the technical challenges of working on Cryptosporidium in the laboratory. Whole genome sequencing and metabolomics have expanded our understanding of the biochemical requirements of this organism and have identified new drug targets. To effectively combat this important pathogen, increased funding is essential.

  11. Bayesian source tracking via focalization and marginalization in an uncertain Mediterranean Sea environment.

    PubMed

    Dosso, Stan E; Wilmut, Michael J; Nielsen, Peter L

    2010-07-01

    This paper applies Bayesian source tracking in an uncertain environment to Mediterranean Sea data, and investigates the resulting tracks and track uncertainties as a function of data information content (number of data time-segments, number of frequencies, and signal-to-noise ratio) and of prior information (environmental uncertainties and source-velocity constraints). To track low-level sources, acoustic data recorded for multiple time segments (corresponding to multiple source positions along the track) are inverted simultaneously. Environmental uncertainty is addressed by including unknown water-column and seabed properties as nuisance parameters in an augmented inversion. Two approaches are considered: Focalization-tracking maximizes the posterior probability density (PPD) over the unknown source and environmental parameters. Marginalization-tracking integrates the PPD over environmental parameters to obtain a sequence of joint marginal probability distributions over source coordinates, from which the most-probable track and track uncertainties can be extracted. Both approaches apply track constraints on the maximum allowable vertical and radial source velocity. The two approaches are applied for towed-source acoustic data recorded at a vertical line array at a shallow-water test site in the Mediterranean Sea where previous geoacoustic studies have been carried out.

  12. Cryptosporidium infections: molecular advances.

    PubMed

    Lendner, Matthias; Daugschies, Arwid

    2014-09-01

    Cryptosporidium host cell interaction remains fairly obscure compared with other apicomplexans such as Plasmodium or Toxoplasma. The reason for this is probably the inability of this parasite to complete its life cycle in vitro and the lack of a system to genetically modify Cryptosporidium. However, there is a substantial set of data about the molecules involved in attachment and invasion and about the host cell pathways involved in actin arrangement that are altered by the parasite. Here we summarize the recent advances in research on host cell infection regarding the excystation process, attachment and invasion, survival in the cell, egress and the available data on omics.

  13. Occurrence of Cryptosporidium sp. oocysts in fecal and water samples in Austria.

    PubMed

    Hassl, A; Benyr, G; Sommer, R

    2001-10-22

    Oocysts of Cryptosporidium spp. were detected and differentiated by a modular arranged gene amplification procedure in various samples, mostly human stool, feces of herpetotaxa, and water, in different locations of South and Eastern Austria. Cryptosporidium parvum was found in stool samples of immunocompromised persons, in reptile feces, and in water samples. The presence of Cryptosporidium in an area is probably associated with high human population densities since water from protected sources in sparsely inhabited areas is rarely contaminated.

  14. A Customized DNA Microarray for Microbial Source Tracking ...

    EPA Pesticide Factsheets

    It is estimated that more than 160, 000 miles of rivers and streams in the United States are impaired due to the presence of waterborne pathogens. These pathogens typically originate from human and other animal fecal pollution sources; therefore, a rapid microbial source tracking (MST) method is needed to facilitate water quality assessment and impaired water remediation. We report a novel qualitative DNA microarray technology consisting of 453 probes for the detection of general fecal and host-associated bacteria, viruses, antibiotic resistance, and other environmentally relevant genetic indicators. A novel data normalization and reduction approach is also presented to help alleviate false positives often associated with high-density microarray applications. To evaluate the performance of the approach, DNA and cDNA was isolated from swine, cattle, duck, goose and gull fecal reference samples, as well as soiled poultry liter and raw municipal sewage. Based on nonmetric multidimensional scaling analysis of results, findings suggest that the novel microarray approach may be useful for pathogen detection and identification of fecal contamination in recreational waters. The ability to simultaneously detect a large collection of environmentally important genetic indicators in a single test has the potential to provide water quality managers with a wide range of information in a short period of time. Future research is warranted to measure microarray performance i

  15. Alternative estimate of source distribution in microbial source tracking using posterior probabilities.

    PubMed

    Greenberg, Joshua; Price, Bertram; Ware, Adam

    2010-04-01

    Microbial source tracking (MST) is a procedure used to determine the relative contributions of humans and animals to fecal microbial contamination of surface waters in a given watershed. Studies of MST methodology have focused on optimizing sampling, laboratory, and statistical analysis methods in order to improve the reliability of determining which sources contributed most to surface water fecal contaminant. The usual approach for estimating a source distribution of microbial contamination is to classify water sample microbial isolates into discrete source categories and calculate the proportion of these isolates in each source category. The set of proportions is an estimate of the contaminant source distribution. In this paper we propose and compare an alternative method for estimating a source distribution-averaging posterior probabilities of source identity across isolates. We conducted a Monte Carlo simulation covering a wide variety of watershed scenarios to compare the two methods. The results show that averaging source posterior probabilities across isolates leads to more accurate source distribution estimates than proportions that follow classification.

  16. Identifying fecal sources in a selected catchment reach using multiple source-tracking tools

    USGS Publications Warehouse

    Vogel, J.R.; Stoeckel, D.M.; Lamendella, R.; Zelt, R.B.; Santo, Domingo J.W.; Walker, S.R.; Oerther, D.B.

    2007-01-01

    Given known limitations of current microbial source-tracking (MST) tools, emphasis on small, simple study areas may enhance interpretations of fecal contamination sources in streams. In this study, three MST tools - Escherichia coli repetitive element polymerase chain reaction (rep-PCR), coliphage typing, and Bacteroidales 16S rDNA host-associated markers - were evaluated in a selected reach of Plum Creek in sooth-central Nebraska. Water-quality samples were collected from six sites. One reach was selected for MST evaluation based on observed patterns of E. coli contamination. Despite high E. coli concentrations, coliphages were detected only once among water samples, precluding their use as a MST tool in this setting. Rep-PCR classification of E. coli isolates from both water and sediment samples supported the hypothesis that cattle and wildlife were dominant sources of fecal contamination, with minor contributions by horses and humans. Conversely, neither ruminant nor human sources were detected by Bacteroidales markers in most water samples. In bed sediment, ruminant- and human-associated Bacteroidales markers were detected throughout the interval from 0 to 0.3 m, with detections independent of E. coli concentrations in the sediment. Although results by E. coli-based and Bacteroidales-based MST methods led to similar interpretations, detection of Bacteroidales markers in sediment more commonly than in water indicates that different tools to track fecal contamination (in this case, tools based on Bacteroidales DNA and E. coli isolates) may have varying relevance to the more specific goal of tracking the sources of E. coli in watersheds. This is the first report of simultaneous, toolbox approach application of a library-based and marker-based MST analyses to lowing surface water. ?? ASA, CSSA, SSSA.

  17. Microbial source tracking: a tool for identifying sources of microbial contamination in the food chain.

    PubMed

    Fu, Ling-Lin; Li, Jian-Rong

    2014-01-01

    The ability to trace fecal indicators and food-borne pathogens to the point of origin has major ramifications for food industry, food regulatory agencies, and public health. Such information would enable food producers and processors to better understand sources of contamination and thereby take corrective actions to prevent transmission. Microbial source tracking (MST), which currently is largely focused on determining sources of fecal contamination in waterways, is also providing the scientific community tools for tracking both fecal bacteria and food-borne pathogens contamination in the food chain. Approaches to MST are commonly classified as library-dependent methods (LDMs) or library-independent methods (LIMs). These tools will have widespread applications, including the use for regulatory compliance, pollution remediation, and risk assessment. These tools will reduce the incidence of illness associated with food and water. Our aim in this review is to highlight the use of molecular MST methods in application to understanding the source and transmission of food-borne pathogens. Moreover, the future directions of MST research are also discussed.

  18. Considering the risk of infection by cryptosporidium via consumption of municipally treated drinking water from a surface water source in a Southwestern Ontario community.

    PubMed

    Pintar, K D M; Fazil, A; Pollari, F; Waltner-Toews, D; Charron, D F; McEwen, S A; Walton, T

    2012-07-01

    Through the use of case-control analyses and quantitative microbial risk assessment (QMRA), relative risks of transmission of cryptosporidiosis have been evaluated (recreational water exposure vs. drinking water consumption) for a Canadian community with higher than national rates of cryptosporidiosis. A QMRA was developed to assess the risk of Cryptosporidium infection through the consumption of municipally treated drinking water. Simulations were based on site-specific surface water contamination levels and drinking water treatment log₁₀ reduction capacity for Cryptosporidium. Results suggested that the risk of Cryptosporidium infection via drinking water in the study community, assuming routine operation of the water treatment plant, was negligible (6 infections per 10¹³ persons per day--5th percentile: 2 infections per 10¹⁵ persons per day; 95th percentile: 3 infections per 10¹² persons per day). The risk is essentially nonexistent during optimized, routine treatment operations. The study community achieves between 7 and 9 log₁₀ Cryptosporidium oocyst reduction through routine water treatment processes. Although these results do not preclude the need for constant vigilance by both water treatment and public health professionals in this community, they suggest that the cause of higher rates of cryptosporidiosis are more likely due to recreational water contact, or perhaps direct animal contact. QMRA can be successfully applied at the community level to identify data gaps, rank relative public health risks, and forecast future risk scenarios. It is most useful when performed in a collaborative way with local stakeholders, from beginning to end of the risk analysis paradigm.

  19. Application of enteric viruses for fecal pollution source tracking in environmental waters

    EPA Science Inventory

    Microbial source tracking (MST) tools are used to identify sources of fecal pollution for accurately assessing public health risk and implementing best management practices (BMPs). This review focuses on the potential of enteric viruses for MST applications. Following host infect...

  20. How can the UK statutory Cryptosporidium monitoring be used for Quantitative Risk Assessment of Cryptosporidium in drinking water?

    PubMed

    Smeets, P W M H; van Dijk, J C; Stanfield, G; Rietveld, L C; Medema, G J

    2007-01-01

    Quantitative Microbiological Risk Assessment (QMRA) is increasingly being used to complement traditional verification of drinking water safety through the absence of indicator bacteria. However, the full benefit of QMRA is often not achieved because of a lack of appropriate data on the fate and behaviour of pathogens. In the UK, statutory monitoring for Cryptosporidium has provided a unique dataset of pathogens directly measured in large volumes of treated drinking water. Using this data a QMRA was performed to determine the benefits and limitations of such state-of-the-art monitoring for risk assessment. Estimates of the risk of infection at the 216 assessed treatment sites ranged from 10(-6.5) to 10(-2.5) person(-1) d(-1). In addition, Cryptosporidium monitoring data in source water was collected at eight treatment sites to determine how Cryptosporidium removal could be quantified for QMRA purposes. Cryptosporidium removal varied from 1.8 to 5.2 log units and appeared to be related to source water Cryptosporidium concentration. Application of general removal credits can either over- or underestimate Cryptosporidium removal by full-scale sedimentation and filtration. State-of-the-art pathogen monitoring can identify poorly performing systems, although it is ineffective to verify drinking water safety to the level of 10(-4) infections person(-1) yr(-1).

  1. Tracking the Sources of Fecal Contaminations: an Interdisciplinary Toolbox

    NASA Astrophysics Data System (ADS)

    Jeanneau, L.; Jarde, E.; Derrien, M.; Gruau, G.; Solecki, O.; Pourcher, A.; Marti, R.; Wéry, N.; Caprais, M.; Gourmelon, M.; Mieszkin, S.; Jadas-Hécart, A.; Communal, P.

    2011-12-01

    Fecal contaminations of inland and coastal waters induce risks to human health and economic losses. In order to improve water management, it is necessary to identify the sources of contamination, which implies the development of specific markers. In order to be considered as a valuable host-specific marker, one must (1) be source specific, (2) occur in high concentration in polluting matrices, (3) exhibit extra-intestinal persistence similar to fecal indicator bacteria (FIB) and (4) not grow out of the host. However, up to day no single marker has fulfilled all those criteria. Thus, it has been suggested to use a combination of markers in order to generate more reliable data. This has lead to the development of a Microbial Source Tracking (MST) toolbox including FIB and microbial and chemical specific markers in order to differentiate between human, bovine and porcine fecal contaminations. Those specific markers are, (1) genotypes of F-specific RNA bacteriophages, (2) bacterial markers belonging to the Bacteroidales (human-specific HF183, ruminant-specific Rum-2-Bac and pig-specific Pig-2-Bac markers), to the Bifidobacterium (Bifidobacterium adolescentis) and pig-specific Lactobacillus amylovorus, (3) fecal stanols and (4) caffeine. The development of this MST toolbox was composed of four steps, from the molecular scale to the watershed scale. At the molecular scale, the specificity and the concentration of those markers were studied in cattle and pig manures and in waste water treatment plant (WWTP) effluents and influents. At the microcosm scale, the transfer of bovine and porcine specific markers was investigated by rainfall simulations on agricultural plots amended with cattle or pig manure. Moreover, the relative persistence of FIB and human, porcine and bovine specific markers was investigated in freshwater and seawater microcosms inoculated with a WWTP influent, pig manure and cow manure. Finally, the aforementioned MST toolbox has been validated at the

  2. Microbial source tracking and transfer hydrodynamics in rural catchments.

    NASA Astrophysics Data System (ADS)

    Murphy, Sinead; Bhreathnach, Niamh; O'Flaherty, Vincent; Jordan, Philip; Wuertz, Stefan

    2013-04-01

    In Ireland, bacterial pathogens from continual point source pollution and intermittent pollution from diffuse sources can impact both drinking water supplies and recreational waters. This poses a serious public health threat. Observing and establishing the source of faecal pollution is imperative for the protection of water quality and human health. Traditional culture methods to detect such pollution via faecal indicator bacteria have been widely utilised but do not decipher the source of pollution. To combat this, microbial source tracking, an important emerging molecular tool, is applied to detect host-specific markers in faecally contaminated waters. The aim of this study is to target ruminant and human-specific faecal Bacteroidales and Bacteroides 16S rRNA genes within rural river catchments in Ireland and investigate hydrological transfer dependencies. During storm events and non-storm periods, 1L untreated water samples, taken every 2 hours over a 48-hour time period at the spring (Cregduff) or outlet (Dunleer), and large (5-20L) untreated water samples were collected from two catchment sites. Cregduff is a spring emergence under a grassland karst landscape in Co. Mayo (west coast of Ireland) and Dunleer is a mixed landuse over till soils in Co. Louth (east coast). From a risk assessment point of view, the catchments are very different. Samples were filtered through 0.2µm nitrocellulose filters to concentrate bacterial cells which then underwent chemical extraction of total nucleic acids. Animal and human stool samples were also collected from the catchments to determine assay sensitivity and specificity following nucleic acid extraction. Aquifer response to seasonal events was assessed by monitoring coliforms and E. coli occurrence using the IDEXX Colisure® Quanti Tray®/2000 system in conjunction with chemical and hydrological parameters. Autoanalysers deployed at each catchment monitor multiple water parameters every 10 min such as phosphorus, nitrogen

  3. A novel microbial source tracking microarray for pathogen detection and fecal source identification in environmental systems.

    PubMed

    Li, Xiang; Harwood, Valerie J; Nayak, Bina; Staley, Christopher; Sadowsky, Michael J; Weidhaas, Jennifer

    2015-06-16

    Pathogen detection and the identification of fecal contamination sources are challenging in environmental waters. Factors including pathogen diversity and ubiquity of fecal indicator bacteria hamper risk assessment and remediation of contamination sources. A custom microarray targeting pathogens (viruses, bacteria, protozoa), microbial source tracking (MST) markers, and antibiotic resistance genes was tested against DNA obtained from whole genome amplification (WGA) of RNA and DNA from sewage and animal (avian, cattle, poultry, and swine) feces. Perfect and mismatch probes established the specificity of the microarray in sewage, and fluorescence decrease of positive probes over a 1:10 dilution series demonstrated semiquantitative measurement. Pathogens, including norovirus, Campylobacter fetus, Helicobacter pylori, Salmonella enterica, and Giardia lamblia were detected in sewage, as well as MST markers and resistance genes to aminoglycosides, beta-lactams, and tetracycline. Sensitivity (percentage true positives) of MST results in sewage and animal waste samples (21-33%) was lower than specificity (83-90%, percentage of true negatives). Next generation DNA sequencing revealed two dominant bacterial families that were common to all sample types: Ruminococcaceae and Lachnospiraceae. Five dominant phyla and 15 dominant families comprised 97% and 74%, respectively, of sequences from all fecal sources. Phyla and families not represented on the microarray are possible candidates for inclusion in subsequent array designs.

  4. Tracking Coherent Structures and Source Localization in Geophysical Flows

    NASA Astrophysics Data System (ADS)

    Forgoston, Eric; Hsieh, Ani; Schwartz, Ira; Yecko, Philip

    There has been a steady increase in the deployment of autonomous underwater and surface vehicles for applications such as ocean monitoring, tracking of marine processes, and forecasting contaminant transport. The underwater environment poses unique challenges since robots must operate in a communication and localization-limited environment where their dynamics are tightly coupled with the environmental dynamics. This work presents current efforts in understanding the impact of geophysical fluid dynamics on underwater vehicle control and autonomy. The focus of the talk is on the use of collaborative vehicles to track Lagrangian coherent structures and to localize contaminant spills. Research supported by the National Science Foundation and the Office of Naval Research.

  5. Evaluating Cryptosporidium and Giardia concentrations in combined sewer overflow.

    PubMed

    Arnone, Russell D; Walling, Joyce Perdek

    2006-06-01

    Since the first identified Cryptosporidium outbreaks occurred in the 1980s and the massive 1993 Milwaukee, WI outbreak affected more than 400,000 people, the concern over the public health risks linked to protozoan pathogens Cryptosporidium and Giardia has grown. Cryptosporidium and Giardia, found in streams, rivers, groundwater, and soil, form hardy, disinfection-resistant oocysts and cysts. Both organisms are recognized causative agents of gastrointestinal illnesses linked to the consumption of contaminated surface or groundwater. This study, the first in a planned series to estimate the urban contribution to the total Cryptosporidium and Giardia receiving-water loads, focused on combined sewer overflow (CSO). CSOs are discharges of mixed untreated sewage and stormwater released directly into receiving waters during rainfall. This engineered relief is necessary to accommodate hydraulic strain when the combined rain and sanitary flows exceed the system capacity. Limited comprehensive data are available assessing the CSO discharge contribution as a source of these two pathogens. Works by States et al. and Gibson et al. each found Cryptosporidium and much greater Giardia concentrations in CSOs draining parts of Pittsburgh, PA. This project estimated the relative detection frequency and concentration of Cryptosporidium and Giardia in CSO. Analytical results were obtained using a modification of Method 1623, originally developed for much cleaner environmental samples. These data are useful for drinking water treatment plants located downstream of CSOs. It is also significant in determining the potential concentrations of parasites at treatment plant intakes and for assessing health risks for water contact and fishing activities. Commonly monitored indicator organisms (total coliform, fecal coliform, E. coli, Enterococcus, and fecal streptococcus), endospores, and selected physical and chemical parameters were analyzed to further describe the samples. CSO from urban

  6. Source tracking fecal bacteria in water: a critical review of current methods.

    PubMed

    Meays, Cynthia L; Broersma, Klaas; Nordin, Rick; Mazumder, Asit

    2004-10-01

    Many molecular and biochemical methods and techniques are being developed to track sources of bacteria in water and food. Currently, there is no standard method proposed for source tracking. This manuscript is a critical evaluation of the various methods used in watersheds, and highlights some of the advantages and disadvantages of each method. Making a decision on a single or combination of methods to use under a particular situation will depend on a number of factors including: question(s) to be answered, scale of identification (broad scale versus specific species identification), available expertise, cost of analysis, turnaround time, and access to facilities. This manuscript reviews several source tracking methodologies which are in current use for source tracking fecal bacteria in the environment including: ribotyping, pulse-field gel electrophoresis, denaturing-gradient gel electrophoresis, repetitive DNA sequences (Rep-PCR), host-specific 16S rDNA genetic markers, and antibiotic resistance analysis.

  7. Techniques for Tracking, Evaluating, and Reporting the Implementation of Nonpoint Source Control Measures - Forestry

    EPA Pesticide Factsheets

    This guidance is intended to assist state, regional, and local environmental professionals in tracking the implementation of best management practices (BMPs) used to control nonpoint source pollution generated by forestry practices.

  8. System and method for tracking a signal source. [employing feedback control

    NASA Technical Reports Server (NTRS)

    Mogavero, L. N.; Johnson, E. G.; Evans, J. M., Jr.; Albus, J. S. (Inventor)

    1978-01-01

    A system for tracking moving signal sources is disclosed which is particularly adaptable for use in tracking stage performers. A miniature transmitter is attached to the person or object to be tracked and emits a detectable signal of a predetermined frequency. A plurality of detectors positioned in a preset pattern sense the signal and supply output information to a phase detector which applies signals representing the angular orientation of the transmitter to a computer. The computer provides command signals to a servo network which drives a device such as a motor driven mirror reflecting the beam of a spotlight, to track the moving transmitter.

  9. Method and apparatus for acquisition and tracking of light sources in a transient event rich environment

    NASA Technical Reports Server (NTRS)

    Kissh, Frank (Inventor); Flynn, David (Inventor); Fowski, Walter (Inventor); Abreu, Rene (Inventor); Miklus, Kenneth (Inventor); Bolin, Kenneth (Inventor)

    1993-01-01

    A method and apparatus for tracking a light source in a transient event rich environment locks on to a light source incident on a field-of-view 1 of a charge-coupled-device (CCD) array 6, validates the permanence of said light source and transmits data relating to the brilliance and location of said light source if said light source is determined to be permanent.

  10. TrackNTrace: A simple and extendable open-source framework for developing single-molecule localization and tracking algorithms.

    PubMed

    Stein, Simon Christoph; Thiart, Jan

    2016-11-25

    Super-resolution localization microscopy and single particle tracking are important tools for fluorescence microscopy. Both rely on detecting, and tracking, a large number of fluorescent markers using increasingly sophisticated computer algorithms. However, this rise in complexity makes it difficult to fine-tune parameters and detect inconsistencies, improve existing routines, or develop new approaches founded on established principles. We present an open-source MATLAB framework for single molecule localization, tracking and super-resolution applications. The purpose of this software is to facilitate the development, distribution, and comparison of methods in the community by providing a unique, easily extendable plugin-based system and combining it with a novel visualization system. This graphical interface incorporates possibilities for quick inspection of localization and tracking results, giving direct feedback of the quality achieved with the chosen algorithms and parameter values, as well as possible sources for errors. This is of great importance in practical applications and even more so when developing new techniques. The plugin system greatly simplifies the development of new methods as well as adapting and tailoring routines towards any research problem's individual requirements. We demonstrate its high speed and accuracy with plugins implementing state-of-the-art algorithms and show two biological applications.

  11. TrackNTrace: A simple and extendable open-source framework for developing single-molecule localization and tracking algorithms

    PubMed Central

    Stein, Simon Christoph; Thiart, Jan

    2016-01-01

    Super-resolution localization microscopy and single particle tracking are important tools for fluorescence microscopy. Both rely on detecting, and tracking, a large number of fluorescent markers using increasingly sophisticated computer algorithms. However, this rise in complexity makes it difficult to fine-tune parameters and detect inconsistencies, improve existing routines, or develop new approaches founded on established principles. We present an open-source MATLAB framework for single molecule localization, tracking and super-resolution applications. The purpose of this software is to facilitate the development, distribution, and comparison of methods in the community by providing a unique, easily extendable plugin-based system and combining it with a novel visualization system. This graphical interface incorporates possibilities for quick inspection of localization and tracking results, giving direct feedback of the quality achieved with the chosen algorithms and parameter values, as well as possible sources for errors. This is of great importance in practical applications and even more so when developing new techniques. The plugin system greatly simplifies the development of new methods as well as adapting and tailoring routines towards any research problem’s individual requirements. We demonstrate its high speed and accuracy with plugins implementing state-of-the-art algorithms and show two biological applications. PMID:27885259

  12. Zoonotic Cryptosporidium spp. and Enterocytozoon bieneusi in pet chinchillas (Chinchilla lanigera) in China.

    PubMed

    Qi, Meng; Luo, Nannan; Wang, Haiyan; Yu, Fuchang; Wang, Rongjun; Huang, Jianying; Zhang, Longxian

    2015-10-01

    Cryptosporidium and Enterocytozoon bieneusi are the most prevalent protist pathogens responsible for inducing human and animal diseases worldwide. The aim of the present work was to determine the occurrence of Cryptosporidium spp. and E. bieneusi in pet chinchillas in China. One hundred forty fecal samples were collected from four cities: Beijing, Zhengzhou, Anyang and Guiyang. They were then examined with PCR amplification of the small subunit ribosomal RNA (SSU rRNA) of Cryptosporidium spp. and the internal transcribed spacer (ITS) of the ribosomal RNA of E. bieneusi. The infection rates for Cryptosporidium spp. and E. bieneusi were 10.0% and 3.6%, respectively. Sequence analysis of SSU rRNA gene products identified two Cryptosporidium spp., Cryptosporidium ubiquitum (n=13) and Cryptosporidium parvum (n=1). Subtyping with the 60-kDa glycoprotein (gp60) gene showed that all C. ubiquitum isolates belonged to zoonotic subtype family XIId, while the subtype of the C. parvum isolate could not be identified. Two E. bieneusi genotypes were identified in five samples, zoonotic genotypes BEB6 (n=3) and D (n=2). This is the first report of C. ubiquitum and C. parvum, and E. bieneusi in chinchillas. This result indicates that pet chinchillas may be a potential source of human infection with Cryptosporidium spp. and E. bieneusi.

  13. Detection of Cryptosporidium sp. Oocyst and Giardia sp. cyst in faucet water samples from cattle and goat farms in Taiwan.

    PubMed

    Watanabe, Yuko; Kimura, Kenji; Yang, Cheng-Hsiung; Ooi, Hong-Kean

    2005-12-01

    A survey on the presence of Cryptosporidium oocyst and Giardia cyst in livestock drinking water as well as the urban tap water throughout Taiwan was carried out. Water examination for the presence of the protozoa was conducted by filtering through a PTFE membrane followed by immunomagnetic separation (IMS) and immunostaining the sediment with commercially available monoclonal antibody against Cryptosporidium and Giardia. Of the 55 different water samples from various sources examined, 2 were found to contain both of Cryptosporidium oocyst and Giardia cyst, 1 was found to contain Cryptosporidium oocyst only. These protozoa-positive water samples, originating from underground well and from the mountain spring, were also used as drinking water for livestock. However, no Cryptosporidium oocyst was found in the city tap water. This is the first report of Cryptosporidium oocyst and Giardia cyst being found in the drinking water for livestock.

  14. Exploring global Cryptosporidium emissions to surface water.

    PubMed

    Hofstra, N; Bouwman, A F; Beusen, A H W; Medema, G J

    2013-01-01

    The protozoan parasite Cryptosporidium is a major cause of diarrhoea worldwide. This paper presents the first model-based inventory with 0.5 by 0.5 degree resolution of global Cryptosporidium emissions for the year 2000 from humans and animals to surface water. The model is based on nutrient distribution modelling, because the sources and transport of oocysts and nutrients to the surface water are comparable. Total emissions consist of point source emissions from wastewater and nonpoint source emissions by runoff of oocysts in manure from agricultural lands. Results indicate a global emission of 3 × 10(17) oocysts per year, with comparable contributions from point and nonpoint sources. Hot-spot areas for point sources are big cities in China, India and Latin America, while the area with the largest nonpoint source emissions is in China. Uncertainties in the model are large. Main areas for further study are (i) excretion rates of oocysts by humans and animals, (ii) emissions of humans not connected to sewage systems, and (iii) retention of oocysts to determine surface water pathogen concentrations rather than emissions. Our results are useful to health organisations to identify priority areas for further study and intervention.

  15. BACTERIA SOURCE TRACKING AND HOST SPECIES SPECIFICITY ANALYSIS

    EPA Science Inventory

    Point and non-point pollution sources of fecal pollution on a watershed adversely impact the quality of drinking source waters and recreational waters. States are required to develop total maximum daily loads (TMDLs) and devise best management practices (BMPs) to reduce the pollu...

  16. DNA BASED MOLECULAR METHODS FOR BACTERIAL SOURCE TRACKING IN WATERSHEDS

    EPA Science Inventory

    Point and non-point pollution sources of fecal pollution on a watershed adversely impact the quality of drinking source waters and recreational waters. States are required to develop total maximum daily loads (TMDLs) and devise best management practices (BMPs) to reduce the po...

  17. Tracking nonpoint source nitrogen pollution in human-impacted watersheds

    USGS Publications Warehouse

    Kaushal, Sujay S.; Groffman, Peter M; Band, Lawrence; Elliott, Emily M.; Shields, Catherine A.; Kendall, Carol

    2011-01-01

    Nonpoint source nitrogen (N) pollution is a leading contributor to U.S. water quality impairments. We combined watershed N mass balances and stable isotopes to investigate fate and transport of nonpoint N in forest, agricultural, and urbanized watersheds at the Baltimore Long-Term Ecological Research site. Annual N retention was 55%, 68%, and 82% for agricultural, suburban, and forest watersheds, respectively. Analysis of δ15N-NO3–, and δ18O-NO3– indicated wastewater was an important nitrate source in urbanized streams during baseflow. Negative correlations between δ15N-NO3– and δ18O-NO3– in urban watersheds indicated mixing between atmospheric deposition and wastewater, and N source contributions changed with storm magnitude (atmospheric sources contributed ∼50% at peak storm N loads). Positive correlations between δ15N-NO3– and δ18O-NO3– in watersheds suggested denitrification was removing septic system and agriculturally derived N, but N from belowground leaking sewers was less susceptible to denitrification. N transformations were also observed in a storm drain (no natural drainage network) potentially due to organic carbon inputs. Overall, nonpoint sources such as atmospheric deposition, wastewater, and fertilizer showed different susceptibility to watershed N export. There were large changes in nitrate sources as a function of runoff, and anticipating source changes in response to climate and storms will be critical for managing nonpoint N pollution.

  18. Tracking nonpoint source nitrogen pollution in human-impacted watersheds.

    PubMed

    Kaushal, Sujay S; Groffman, Peter M; Band, Lawrence E; Elliott, Emily M; Shields, Catherine A; Kendall, Carol

    2011-10-01

    Nonpoint source nitrogen (N) pollution is a leading contributor to U.S. water quality impairments. We combined watershed N mass balances and stable isotopes to investigate fate and transport of nonpoint N in forest, agricultural, and urbanized watersheds at the Baltimore Long-Term Ecological Research site. Annual N retention was 55%, 68%, and 82% for agricultural, suburban, and forest watersheds, respectively. Analysis of δ(15)N-NO(3)(-), and δ(18)O-NO(3)(-) indicated wastewater was an important nitrate source in urbanized streams during baseflow. Negative correlations between δ(15)N-NO(3)(-) and δ(18)O-NO(3)(-) in urban watersheds indicated mixing between atmospheric deposition and wastewater, and N source contributions changed with storm magnitude (atmospheric sources contributed ∼50% at peak storm N loads). Positive correlations between δ(15)N-NO(3)(-) and δ(18)O-NO(3)(-) in watersheds suggested denitrification was removing septic system and agriculturally derived N, but N from belowground leaking sewers was less susceptible to denitrification. N transformations were also observed in a storm drain (no natural drainage network) potentially due to organic carbon inputs. Overall, nonpoint sources such as atmospheric deposition, wastewater, and fertilizer showed different susceptibility to watershed N export. There were large changes in nitrate sources as a function of runoff, and anticipating source changes in response to climate and storms will be critical for managing nonpoint N pollution.

  19. Surveillance Systems for Waterborne Cryptosporidium: US EPA method 1523 and Beyond

    EPA Science Inventory

    Waterborne cryptosporidiosis remains a significant public health concern in countries around the world. Many species and genotypes of Cryptosporidium contaminate drinking water sources, but C. parvum and C. hominis remain the two predominant species known to cause waterborne dis...

  20. FECAL SOURCE TRACKING BY ANTIBIOTIC RESISTANCE ANALYSIS ON A WATERSHED EXHIBITING LOW RESISTANCE

    EPA Science Inventory

    The ongoing development of microbial source tracking has made it possible to identify contamination sources with varying accuracy, depending on the method used. The purpose of this study was done to test the efficiency of the antibiotic resistance analysis (ARA) method under low ...

  1. A Customized DNA Microarray for Microbial Source Tracking in Environmental Systems

    EPA Science Inventory

    It is estimated that more than 160, 000 miles of rivers and streams in the United States are impaired due to the presence of waterborne pathogens. These pathogens typically originate from human and other animal fecal pollution sources; therefore, a rapid microbial source tracking...

  2. A COMPARISON OF ENUMERATION TECHNIQUES FOR CRYPTOSPORIDIUM PARVUM OOCYSTS

    EPA Science Inventory

    A variety of methods have been used to enumerate Cryptosporidium parvum oocysts from source or drinking waters. The reliability of these counting methods varies, in part, with suspension density, sample purity, and other factors. Frequently, the method of determination of suspens...

  3. UTILITY OF SURROGATES FOR MEASURING CRYPTOSPORIDIUM OOCYST INFECTIVITY

    EPA Science Inventory

    The water industry must assess whether Cryptosporidium oocysts detected in source and finished water are viable and/or infectious. Initial approaches measuring the infectious nature of C. parvum oocysts have focused on in vitro excystation and in vitro vital dye staining. Recen...

  4. UTILITY OF SURROGATES FOR MEASURING CRYPTOSPORIDIUM OOCYSTS INFECTIVITY

    EPA Science Inventory

    The water industry must assess whether Cryptosporidium oocysts detected in source and finished water are viable and/or infectious. Initial approaches measuring the infectious nature of C. parvum oocysts have focused on in vitro excystation and in vitro vital dye staining. Recen...

  5. Molecular characterisation of Cryptosporidium and Giardia in cats (Felis catus) in Western Australia.

    PubMed

    Yang, Rongchang; Ying, Joyce Lau Jie; Monis, Paul; Ryan, Una

    2015-08-01

    Little is known of the prevalence of Cryptosporidium and Giardia in domestic cats in Western Australia and their potential role as zoonotic reservoirs for human infection. In the present study, a total of 345 faecal samples from four different sources were screened for the presence of Cryptosporidium and Giardia by PCR and genotyped by sequence analysis. Oocyst numbers and cyst numbers for Cryptosporidium and Giardia respectively were also determined using quantitative PCR assays. Cryptosporidium and Giardia were detected in 9.9% (95% CI 6.7-13.0) and 10.1% (95% CI 7.0-13.3) of cats in Western Australia respectively. Sequence analysis at the 18S rRNA locus identified five Cryptosporidium species/genotypes; C. felis (n = 8), C. muris (n = 1), C. ryanae (n = 1), Cryptosporidium rat genotype III (n = 5) and a novel genotype most closely related to Cryptosporidium rat genotype III in one isolate. This is the first report of C. ryanae and Cryptosporidium rat genotype III in cats. For Giardia, assemblage F the most commonly identified species, while only 1 assemblage sequence was detected. Since most human cases of cryptosporidiosis are caused by C. parvum and C. hominis and human cases of giardiasis are caused by G. duodenalis assemblage A and B, the domestic cats in the present study are likely to be of low zoonotic risk to pet owners in Perth. Risk analyses identified that elderly cats (more than 6 years) were more prone to Cryptosporidium and Giardia infections than kittens (less than 6 months) (P = 0.009). Clinical symptoms were not associated with the prevalence of Cryptosporidium and Giardia infections in cats.

  6. RadSTraM: Radiological Source Tracking and Monitoring, Phase II Final Report

    SciTech Connect

    Warren, Tracy A; Walker, Randy M; Hill, David E; Gross, Ian G; Smith, Cyrus M; Abercrombie, Robert K

    2008-12-01

    This report focuses on the technical information gained from the Radiological Source Tracking and Monitoring (RadSTraM) Phase II investigation and its implications. The intent of the RadSTraM project was to determine the feasibility of tracking radioactive materials in commerce, particularly International Atomic Energy Agency (IAEA) Category 3 and 4 materials. Specifically, Phase II of the project addressed tracking radiological medical isotopes in commerce. These categories of materials are susceptible to loss or theft but the problem is not being addressed by other agencies.

  7. Molecular Epidemiologic Source Tracking of Orally Transmitted Chagas Disease, Venezuela

    PubMed Central

    Segovia, Maikell; Martínez, Clara E.; Messenger, Louisa A.; Nessi, Anaibeth; Londoño, Juan C.; Espinosa, Raul; Martínez, Cinda; Alfredo, Mijares; Bonfante-Cabarcas, Rafael; Lewis, Michael D.; de Noya, Belkisyolé A.; Miles, Michael A.; Llewellyn, Martin S.

    2013-01-01

    Oral outbreaks of Chagas disease are increasingly reported in Latin America. The transitory presence of Trypanosoma cruzi parasites within contaminated foods, and the rapid consumption of those foods, precludes precise identification of outbreak origin. We report source attribution for 2 peri-urban oral outbreaks of Chagas disease in Venezuela via high resolution microsatellite typing. PMID:23768982

  8. Sparsity-driven Passive Tracking of Underwater Acoustic Sources

    DTIC Science & Technology

    2015-08-01

    bend ✴ Temperature , pressure and salinity D epth Range Source localization map (SLM) 2 Shallow water sound -speed profile Modeling...Difficult due to complexities of the propagation environment • Ocean behaves as an acoustic waveguide • Varying sound -speed causes acoustic signals to

  9. Molecular epidemiologic source tracking of orally transmitted Chagas disease, Venezuela.

    PubMed

    Segovia, Maikell; Carrasco, Hernán J; Martínez, Clara E; Messenger, Louisa A; Nessi, Anaibeth; Londoño, Juan C; Espinosa, Raul; Martínez, Cinda; Alfredo, Mijares; Bonfante-Cabarcas, Rafael; Lewis, Michael D; de Noya, Belkisyolé A; Miles, Michael A; Llewellyn, Martin S

    2013-07-01

    Oral outbreaks of Chagas disease are increasingly reported in Latin America. The transitory presence of Trypanosoma cruzi parasites within contaminated foods, and the rapid consumption of those foods, precludes precise identification of outbreak origin. We report source attribution for 2 peri-urban oral outbreaks of Chagas disease in Venezuela via high resolution microsatellite typing.

  10. The Cryptosporidium parvum Kinome

    PubMed Central

    2011-01-01

    Background Hundreds of millions of people are infected with cryptosporidiosis annually, with immunocompromised individuals suffering debilitating symptoms and children in socioeconomically challenged regions at risk of repeated infections. There is currently no effective drug available. In order to facilitate the pursuit of anti-cryptosporidiosis targets and compounds, our study spans the classification of the Cryptosporidium parvum kinome and the structural and biochemical characterization of representatives from the CDPK family and a MAP kinase. Results The C. parvum kinome comprises over 70 members, some of which may be promising drug targets. These C. parvum protein kinases include members in the AGC, Atypical, CaMK, CK1, CMGC, and TKL groups; however, almost 35% could only be classified as OPK (other protein kinases). In addition, about 25% of the kinases identified did not have any known orthologues outside of Cryptosporidium spp. Comparison of specific kinases with their Plasmodium falciparum and Toxoplasma gondii orthologues revealed some distinct characteristics within the C. parvum kinome, including potential targets and opportunities for drug design. Structural and biochemical analysis of 4 representatives of the CaMK group and a MAP kinase confirms features that may be exploited in inhibitor design. Indeed, screening CpCDPK1 against a library of kinase inhibitors yielded a set of the pyrazolopyrimidine derivatives (PP1-derivatives) with IC50 values of < 10 nM. The binding of a PP1-derivative is further described by an inhibitor-bound crystal structure of CpCDPK1. In addition, structural analysis of CpCDPK4 identified an unprecedented Zn-finger within the CDPK kinase domain that may have implications for its regulation. Conclusions Identification and comparison of the C. parvum protein kinases against other parasitic kinases shows how orthologue- and family-based research can be used to facilitate characterization of promising drug targets and the search

  11. Degradation and Isotope Source Tracking of Glyphosate and Aminomethylphosphonic Acid.

    PubMed

    Li, Hui; Joshi, Sunendra R; Jaisi, Deb P

    2016-01-27

    Glyphosate [N-(phosphonomethyl) glycine], an active ingredient of the herbicide Roundup, and its main metabolite, aminomethylphosphonic acid (AMPA), have been frequently reported to be present in soils and other environments and thus have heightened public concerns on their potential adverse effects. Understanding the fate of these compounds and differentiating them from other naturally occurring compounds require a toolbox of methods that can go beyond conventional methods. Here, we applied individual isotope labeling technique whereby each compound or mineral involved in the glyphosate and AMPA degradation reaction was either synthesized or chosen to have distinct (18)O/(16)O ratios so that the source of incorporated oxygen in the orthophosphate generated and corresponding isotope effect during C-P bond cleavage could be identified. Furthermore, we measured original isotope signatures of a few commercial glyphosate sources to identify their source-specific isotope signatures. Our degradation kinetics results showed that the rate of glyphosate degradation was higher than that of AMPA in all experimental conditions, and both the rate and extent of degradation were lowest under anoxic conditions. Oxygen isotope ratios (δ(18)OP) of orthophosphate generated from glyphosate and AMPA degradation suggested that one external oxygen atom from ambient water, not from dissolved oxygen or mineral, was incorporated into orthophosphate with the other three oxygen atoms inherited from the parent molecule. Interestingly, δ(18)OP values of all commercial glyphosate products studied were found to be the lightest among all orthophosphates known so far. Furthermore, isotope composition was found to be unaffected due to variable degradation kinetics, light/dark, and oxic/anoxic conditions. These results highlight the importance of phosphate oxygen isotope ratios as a nonconventional tool to potentially distinguish glyphosate sources and products from other organophosphorus compounds

  12. Prevalence of Cryptosporidium and Giardia in the water resources of the Kuang River catchment, Northern Thailand.

    PubMed

    Chuah, C Joon; Mukhaidin, Nabila; Choy, Seow Huey; Smith, Gavin J D; Mendenhall, Ian H; Lim, Yvonne A L; Ziegler, Alan D

    2016-08-15

    A catchment-scale investigation of the prevalence of Cryptosporidium and Giardia in the Kuang River Basin was carried out during the dry and rainy seasons. Water samples were collected from the Kuang River and its tributaries as well as a major irrigation canal at the study site. We also investigated the prevalence of gastrointestinal parasitic infection among dairy and beef cattle hosts. Cryptosporidium and/or Giardia were detected in all the rivers considered for this study, reflecting their ubiquity within the Kuang River Basin. The high prevalence of Cryptosporidium/Giardia in the upper Kuang River and Lai River is of a particular concern as both drain into the Mae Kuang Reservoir, a vital source of drinking-water to many local towns and villages at the research area. We did not, however, detected neither Cryptosporidium nor Giardia were in the irrigation canal. The frequency of Cryptosporidium/Giardia detection nearly doubled during the rainy season compared to the dry season, highlighting the importance of water as an agent of transport. In addition to the overland transport of these protozoa from their land sources (e.g. cattle manure, cess pits), Cryptosporidium/Giardia may also be re-suspended from the streambeds (a potentially important repository) into the water column of rivers during storm events. Faecal samples from dairy and beef cattle showed high infection rates from various intestinal parasites - 97% and 94%, respectively. However, Cryptosporidium and Giardia were only detected in beef cattle. The difference in management style between beef (freeranging) and dairy cattle (confined) may account for this disparity. Finally, phylogenetic analyses revealed that the Cryptosporidium/Giardia-positive samples contained C. ryanae (non-zoonotic) as well as Giardia intestinalis assemblages B (zoonotic) and E (non-zoonotic). With only basic water treatment facilities afforded to them, the communities of the rural area relying on these water supplies are

  13. LT2 Cryptosporidium data: what do they tell us about Cryptosporidium in surface water in the United States?

    PubMed

    Ongerth, Jerry E

    2013-05-07

    Beginning in 2006 a United States Federal regulation required public water suppliers using surface water serving more than 10,000 population to analyze for Cryptosporidium in at least 24 consecutive monthly samples from each surface water source. In July 2012, the U.S. EPA released the resulting data consisting of ca. 45,000 records. No Cryptosporidium were found in 93% of samples and no Cryptosporidium were found in any samples analyzed from over half of 1670 locations sampled. Nevertheless, at 250 locations representing every region of the U.S., Cryptosporidium were found in sufficient numbers of samples to provide a picture of their occurrence nationwide. Data from about 100 sites reporting the highest numbers were examined in detail. Although analysis of matrix spikes was required for quality control, the results do not permit estimating organism concentrations. The data reported at each of the individual sample locations were analyzed in the form of cumulative probability distributions to describe key risk-related features of median level and variability. Taken as a whole, the data describe a spectrum of median Cryptosporidium occurrence in surface waters of the U.S. ranging from ca. 0.005 to ca. 0.5 oocysts/L. The variability at individual sites ranged from ca. 1 to 15 r.s.d. Based on the LT2 positive data, comparison to measurements of other water quality parameters, and independent means of estimating organism production from watersheds reported in the literature, the hypothesis is offered that Cryptosporidium may be found in surface water anywhere worldwide continuously and within the spectrum defined above.

  14. Detection and Molecular Characterization of Cryptosporidium spp. from Wild Rodents and Insectivores in South Korea.

    PubMed

    Song, Juha; Kim, C-Yoon; Chang, Seo-Na; Abdelkader, Tamer Said; Han, Juhee; Kim, Tae-Hyun; Oh, Hanseul; Lee, Ji Min; Kim, Dong-Su; Kim, Jong-Taek; Oh, Hong-Shik; Hur, Moonsuk; Suh, Jae-Hwa; Park, Jae-Hak

    2015-12-01

    In order to examine the prevalence of Cryptosporidium infection in wild rodents and insectivores of South Korea and to assess their potential role as a source of human cryptosporidiosis, a total of 199 wild rodents and insectivore specimens were collected from 10 regions of South Korea and screened for Cryptosporidium infection over a period of 2 years (2012-2013). A nested-PCR amplification of Cryptosporidium oocyst wall protein (COWP) gene fragment revealed an overall prevalence of 34.2% (68/199). The sequence analysis of 18S rRNA gene locus of Cryptosporidium was performed from the fecal and cecum samples that tested positive by COWP amplification PCR. As a result, we identified 4 species/genotypes; chipmunk genotype I, cervine genotype I, C. muris, and a new genotype which is closely related to the bear genotype. The new genotype isolated from 12 Apodemus agrarius and 2 Apodemus chejuensis was not previously identified as known species or genotype, and therefore, it is supposed to be a novel genotype. In addition, the host spectrum of Cryptosporidium was extended to A. agrarius and Crosidura lasiura, which had not been reported before. In this study, we found that the Korean wild rodents and insectivores were infected with various Cryptosporidium spp. with large intra-genotypic variationa, indicating that they may function as potential reservoirs transmitting zoonotic Cryptosporidium to livestock and humans.

  15. Detection and Molecular Characterization of Cryptosporidium spp. from Wild Rodents and Insectivores in South Korea

    PubMed Central

    Song, Juha; Kim, C-Yoon; Chang, Seo-Na; Abdelkader, Tamer Said; Han, Juhee; Kim, Tae-Hyun; Oh, Hanseul; Lee, Ji Min; Kim, Dong-Su; Kim, Jong-Taek; Oh, Hong-Shik; Hur, Moonsuk; Suh, Jae-Hwa; Park, Jae-Hak

    2015-01-01

    In order to examine the prevalence of Cryptosporidium infection in wild rodents and insectivores of South Korea and to assess their potential role as a source of human cryptosporidiosis, a total of 199 wild rodents and insectivore specimens were collected from 10 regions of South Korea and screened for Cryptosporidium infection over a period of 2 years (2012-2013). A nested-PCR amplification of Cryptosporidium oocyst wall protein (COWP) gene fragment revealed an overall prevalence of 34.2% (68/199). The sequence analysis of 18S rRNA gene locus of Cryptosporidium was performed from the fecal and cecum samples that tested positive by COWP amplification PCR. As a result, we identified 4 species/genotypes; chipmunk genotype I, cervine genotype I, C. muris, and a new genotype which is closely related to the bear genotype. The new genotype isolated from 12 Apodemus agrarius and 2 Apodemus chejuensis was not previously identified as known species or genotype, and therefore, it is supposed to be a novel genotype. In addition, the host spectrum of Cryptosporidium was extended to A. agrarius and Crosidura lasiura, which had not been reported before. In this study, we found that the Korean wild rodents and insectivores were infected with various Cryptosporidium spp. with large intra-genotypic variationa, indicating that they may function as potential reservoirs transmitting zoonotic Cryptosporidium to livestock and humans. PMID:26797442

  16. RadTrac : A system for detecting, localizing, and tracking radioactive sources in real time.

    SciTech Connect

    Vilim, R.; Klann, R.; Nuclear Engineering Division

    2009-10-01

    Within the homeland security and emergency response communities, there is a need for a low-profile system to detect and locate radioactive sources. RadTrac has been developed at Argonne National Laboratory as an integrated system for the detection, localization, identification, and tracking of radioactive sources in real time. The system is based on a network of radiation detectors and advanced signal-processing algorithms. Features include video surveillance, automated tracking, easy setup, and logging of all data and images. This paper describes the advanced algorithms that were developed and implemented for source detection, localization, and tracking in real time. In the physio-spatial integration approach to source localization, counts from multiple detectors are processed according to the underlying physics linking these counts to obtain the probability that a source is present at any point in space. This information is depicted in a probability density function map. This type of depiction allows the results to be presented in a simple, easy-to-understand manner. It also allows for many different complicated factors to be accounted for in a single image as each factor is computed as a probability density in space. These factors include spatial limitations, variable shielding, directional detectors, moving detectors, and different detector sizes and orientations. The utility and versatility of this approach is described in further detail. Advanced signal-processing algorithms have also been incorporated to improve real-time tracking and to increase signal-to-noise ratios including temporal linking and energy binning. Measurements aimed at demonstrating the sensitivity improvements through the use of advanced signal-processing techniques were performed and are presented. Results of tracking weak sources (<100 {micro}Ci {sup 137}Cs) using four fixed-position detectors are presented.

  17. Performance of forty-one microbial source tracking methods: A twenty-seven lab evaluation study

    EPA Science Inventory

    The last decade has seen development of numerous new microbial source tracking (MST) methodologies, but many of these have been tested in just a few laboratories with a limited number of fecal samples. This method evaluation study examined the specificity and sensitivity of 43 ...

  18. Signal simulator model A-111. [signal source for Doppler tracking high data rate receivers

    NASA Technical Reports Server (NTRS)

    Flattau, T.; Mellars, J.

    1974-01-01

    A signal simulator designed to function as a signal source for Doppler tracking high data rate receivers is described. The simulator produces modulated signals whose carrier frequency can be varied between 200 and 900 MHz at rates greater than 20 MHz/sec. The modulation is phase shift keying with data rate up to 300 megabits per second.

  19. MICROBIAL SOURCE TRACKING - WHERE ARE WE NOW AND WHERE ARE WE GOING?

    EPA Science Inventory

    Microbial Source Tracking ? Where Are We Now and Where Are We Going?

    J. W. Santo-Domingo1, J. M. Simpson1, G. Scott2 and D. J. Reasoner1
    1U.S. Environmental Protection Agency, Cincinnati, OH 45268
    2National Oceanic and Atmospheric Administration, Charleston, SC 29412...

  20. Linear track estimation using double pulse sources for near-field underwater moving target

    NASA Astrophysics Data System (ADS)

    Chen, Zhifei; Hou, Hong; Yang, Jianhua; Sun, Jincai; Wang, Qian

    2013-06-01

    The double pulse sources (DPS) method is presented for linear track estimation in this work. In the field of noise identification of underwater moving target, the Doppler will distort the frequency and amplitude of the radiated noise. To eliminate this, the track estimation is necessary. In the DPS method, we first estimate bearings of two sinusoidal pulse sources installed in the moving target through baseline positioning method. Meanwhile, the emitted and recorded time of each pulse are also acquired. Then the linear track parameters will be achieved based on the geometry pattern with the help of double sources spacing. The simulated results confirm that the DPS improves the performance of the previous double source spacing method. The simulated experiments were carried out using a moving battery car to further evaluate its performance. When the target is 40-60m away, the experiment results show that biases of track azimuth and abeam distance of DPS are under 0.6o and 3.4m, respectively. And the average deviation of estimated velocity is around 0.25m/s.

  1. New Performance Metrics for Quantitative Polymerase Chain Reaction-Based Microbial Source Tracking Methods

    EPA Science Inventory

    Binary sensitivity and specificity metrics are not adequate to describe the performance of quantitative microbial source tracking methods because the estimates depend on the amount of material tested and limit of detection. We introduce a new framework to compare the performance ...

  2. Decay of Fecal Indicator Bacteria and Microbial Source Tracking Markers in Cattle Feces

    EPA Science Inventory

    The survival of fecal indicator bacteria (FIB) and microbial source tracking (MST) markers in water microcosms and manure amended soils has been well documented; however, little is known about the survival of MST markers in bovine feces deposited on pastures. We conducted a study...

  3. Active microcoil tracking in the lungs using a semisolid rubber as signal source.

    PubMed

    Alt, Stefan; Homagk, Ann-Kathrin; Umathum, Reiner; Semmler, Wolfhard; Bock, Michael

    2010-07-01

    A new method to localize and track medical devices in air-filled body cavities is proposed that uses active microcoils with a semisolid filling. In air spaces, e.g., the lung, microcoils require an independent signal source, which should be made of a biocompatible, solid and sterilizable material with a long shelf time. In a measurement of the T(1) and T*(2) and the relative spin density of several semisolid materials, latex was identified as a suitable material from which a prototype catheter was constructed with a microcoil at its tip. In a dual-echo tracking pulse sequence, the very short T*(2) of the rubber material allowed suppressing the background signal from surrounding tissue with a subtraction technique and additional dephasing gradients. With a roadmapping reconstruction, the microcoil's trajectory could be visualized on a previously acquired reference image set with a tracking rate of up to 60 Hz at a spatial resolution of better than 2mm. In a real-time tracking implementation, an image update rate of 4 Hz was achieved by combining the tracking with a fast real-time imaging sequence. Both methods were successfully applied in vivo to track the catheter in the lung of a pig.

  4. Mobile-phone based visible light communication using region-grow light source tracking for unstable light source.

    PubMed

    Liang, Kevin; Chow, Chi-Wai; Liu, Yang

    2016-07-25

    In order to increase the data rate of the camera-based visible light communication (VLC) system, using rolling shutter effect has been demonstrated successfully, in which the pixel rows of the complementary-metal-oxide-semiconductor (CMOS) image sensor are activated sequentially. Previous camera-based VLCs focused on using a stable LED light source, and its illumination area is positioned at the center of an image frame. In this work, we investigate the performance of a camera-based VLC with light source at different parts of an image frame. We propose and demonstrate using region-grow algorithm to track the light source. We also evaluate and discuss different scenarios when the light source is moved. Besides, a recorded > 5 kbit/s net data rate can be achieved by using only a single phosphor-based white-light LED source. Here, we demonstrate that 4.502 pixel/bit can be achieved.

  5. Global Radiological Source Sorting, Tracking, and Monitoring Project: Phase I Final Report

    SciTech Connect

    Walker, Randy M; Hill, David E; Gorman, Bryan L

    2010-09-01

    As a proof of concept tested in an operational context, the Global Radiological Source Sorting, Tracking, and Monitoring (GRadSSTraM) Project successfully demonstrated that radio frequency identification (RFID) and Web 2.0* technologies can be deployed to track controlled shipments between the United States and the European Union. Between November 2009 and May 2010, a total of 19 shipments were successfully shipped from Oak Ridge National Laboratory (ORNL) by the U.S. Postal Service (USPS) and tracked to their delivery at England's National Physical Laboratory (NPL) by the United Kingdom Royal Mail. However, the project can only be viewed as a qualified success as notable shortcomings were observed. Although the origin and terminus of all RFID-enabled shipments were recorded and no shipments were lost, not all the waypoints between ORNL and NPL were incorporated into the pilot. Given limited resources, the project team was able to install RFID listeners/actuators at three waypoints between the two endpoints. Although it is likely that all shipments followed the same route between ORNL and NPL, it cannot be determined beyond question that all 19 shipments were routed on identical itineraries past the same three waypoints. The pilot also raises the distinct possibility that unattended RFID tracking alone, without positive confirmation that a tagged item has been properly recorded by an RFID reader, does not meet a rigorous standard for shipping controlled items. Indeed, the proof of concept test strongly suggests that a multifaceted approach to tracking may be called for, including tracking methods that are capable of reading and accepting multiple inputs for individual items [e.g., carrier-provided tracking numbers, Universal Product Codes (UPCs), and RFID tags]. For controlled items, another apparent requirement is a confirmation feature, human or otherwise, which can certify that an item's RFID tag, UPC, or tracking number has been recorded.

  6. Beamforming with a volumetric array of massless laser spark sources--Application in reflection tracking.

    PubMed

    Eskelinen, Joona; Hæggström, Edward; Delikaris-Manias, Symeon; Bolaños, Javier Gómez; Pulkki, Ville

    2015-06-01

    A volumetric array of laser-induced air breakdown sparks is used to produce a directional and steerable acoustic source. The laser breakdown array element is broadband, point-like, and massless. It produces an impulse-like waveform in midair, thus generating accurate spatio-temporal information for acoustic beamforming. A laser-spark scanning setup and the concept of a massless steerable source are presented and evaluated with a cubic array by using an off-line far field delay-and-sum beamforming method. This virtual acoustic array with minimal source influence can, for instance, produce narrow transmission beams to obtain localized and directional impulse response information by reflection tracking.

  7. Occurrence of Cryptosporidium oocysts in Wrinkled Hornbill and other birds in the Kuala Lumpur National Zoo.

    PubMed

    Rohela, M; Lim, Y A L; Jamaiah, I; Khadijah, P Y Y; Laang, S T; Nazri, M H Mohd; Nurulhuda, Z

    2005-01-01

    The occurrence of a coccidian parasite, Cryptosporidium, among birds in the Kuala Lumpur National Zoo was investigated in this study. A hundred bird fecal samples were taken from various locations of the zoo. Fecal smears prepared using direct smear and formalin ethyl acetate concentration technique were stained with modified Ziehl-Neelsen stain. Samples positive for Cryptosporidium with Ziehl-Neelsen stain were later confirmed using the immunofluorescence technique and viewed under the epifluorescence microscope. Six species of bird feces were confirmed positive with Cryptosporidium oocysts. They included Wrinkled Hornbill (Aceros corrugatus), Great Argus Pheasant (Argusianus argus), Black Swan (Cygnus atratus), Swan Goose (Anser cygnoides), Marabou Stork (Leptoptilos crumeniferus), and Moluccan Cockatoo (Cacatua moluccencis). These birds were located in the aviary and lake, with the Moluccan Cockatoo routinely used as a show bird. Results obtained in this study indicated that animal sanctuaries like zoos and bird parks are important sources of Cryptosporidium infection to humans, especially children and other animals.

  8. Quantitative detection of Cryptosporidium oocyst in water source based on 18S rRNA by alternately binding probe competitive reverse transcription polymerase chain reaction (ABC-RT-PCR).

    PubMed

    Kishida, Naohiro; Miyata, Ryo; Furuta, Atsushi; Izumiyama, Shinji; Tsuneda, Satoshi; Sekiguchi, Yuji; Noda, Naohiro; Akiba, Michihiro

    2012-01-01

    We describe an assay for simple and cost-effective quantification of Cryptosporidium oocysts in water samples using a recently developed quantification method named alternately binding probe competitive PCR (ABC-PCR). The assay is based on the detection of 18S rRNA specific for Cryptosporidium oocysts. The standard curve of the ABC-PCR assay had a good fitting to a rectangular hyperbola with a correlation coefficient (R) of 0.9997. Concentrations of Cryptosporidium oocysts in real river water samples were successfully quantified by the ABC-reverse transcription (RT)-PCR assay. The quantified values by the ABC-RT-PCR assay very closely resemble those by the real-time RT-PCR assay. In addition, the quantified concentration in most water samples by the ABC-RT-PCR assay was comparable to that by conventional microscopic observation. Thus, Cryptosporidium oocysts in water samples can be accurately and specifically determined by the ABC-RT-PCR assay. As the only equipment that is needed for this end-point fluorescence assay is a simple fluorometer and a relatively inexpensive thermal cycler, this method can markedly reduce time and cost to quantify Cryptosporidium oocysts and other health-related water microorganisms.

  9. Twelve years of tracking 52-Hz whale calls from a unique source in the North Pacific

    NASA Astrophysics Data System (ADS)

    Watkins, William A.; Daher, Mary Ann; George, Joseph E.; Rodriguez, David

    2004-12-01

    A unique whale call with 50-52 Hz emphasis from a single source has been tracked over 12 years in the central and eastern North Pacific. These calls, referred to as 52-Hz calls, were monitored and analyzed from acoustic data recorded by hydrophones of the US Navy Sound Surveillance System (SOSUS) and other arrays. The calls were noticed first in 1989, and have been detected and tracked since 1992. No other calls with similar characteristics have been identified in the acoustic data from any hydrophone system in the North Pacific basin. Only one series of these 52-Hz calls has been recorded at a time, with no call overlap, suggesting that a single whale produced the calls. The calls were recorded from August to February with most in December and January. The species producing these calls is unknown. The tracks of the 52-Hz whale were different each year, and varied in length from 708 to 11,062 km with travel speeds ranging from 0.7 to 3.8 km/h. Tracks included (A) meandering over short ranges, (B) predominantly west-to-east movement, and (C) mostly north-to-south travel. These tracks consistently appeared to be unrelated to the presence or movement of other whale species (blue, fin and humpback) monitored year-round with the same hydrophones.

  10. Sea ice drift from Sentinel-1 SAR imagery using open source feature tracking

    NASA Astrophysics Data System (ADS)

    Muckenhuber, S.; Korosov, A.; Sandven, S.

    2015-12-01

    A computational efficient, open source feature tracking algorithm, called ORB, is adopted and tuned for sea ice drift retrieval from Sentinel-1 SAR images. The best suitable setting and parameter values have been found using four representative Sentinel-1 image pairs. A new quality measure for feature tracking algorithms is introduced utilising the distribution of the resulting vector field. The performance of the algorithm is compared with two other feature tracking algorithms (SIFT and SURF). Applied on a test image pair acquired over Fram Strait, the tuned ORB algorithm produces the highest number of vectors (6920, SIFT: 1585 and SURF: 518) while being computational most efficient (66 s, SIFT: 182 s and SURF: 99 s using a 2,7 GHz processor with 8 GB memory). For validation purpose, 350 manually drawn vectors have been compared with the closest calculated vectors and the resulting root mean square distance is 609.9 m (equivalent to 7.5 pixel). All test image pairs show a significant better performance of the HV channel. On average, around 4 times more vectors have been found using HV polarisation. All software requirements necessary for applying the presented feature tracking algorithm are open source to ensure a free and easy implementation.

  11. Sea ice drift from Sentinel-1 SAR imagery using open source feature tracking

    NASA Astrophysics Data System (ADS)

    Muckenhuber, Stefan; Korosov, Anton; Sandven, Stein

    2016-04-01

    A computational efficient, open source feature tracking algorithm, called ORB, is adopted and tuned for sea ice drift retrieval from Sentinel-1 SAR images. The best suitable setting and parameter values have been found using four representative Sentinel-1 image pairs. A new quality measure for feature tracking algorithms is introduced utilising the distribution of the resulting vector field. The performance of the algorithm is compared with two other feature tracking algorithms (SIFT and SURF). Applied on a test image pair acquired over Fram Strait, the tuned ORB algorithm produces the highest number of vectors (6920, SIFT: 1585 and SURF: 518) while being computational most efficient (66 s, SIFT: 182 s and SURF: 99 s using a 2,7 GHz processor with 8 GB memory). For validation purpose, 350 manually drawn vectors have been compared with the closest calculated vectors and the resulting root mean square distance is 609.9 m (equivalent to 7.5 pixel). All test image pairs show a significant better performance of the HV channel. On average, around 4 times more vectors have been found using HV polarisation. All software requirements necessary for applying the presented feature tracking algorithm are open source to ensure a free and easy implementation.

  12. Zoonotic Cryptosporidium Species in Animals Inhabiting Sydney Water Catchments

    PubMed Central

    Zahedi, Alireza; Monis, Paul; Aucote, Sarah; King, Brendon; Paparini, Andrea; Jian, Fuchun; Yang, Rongchang; Oskam, Charlotte; Ball, Andrew; Robertson, Ian; Ryan, Una

    2016-01-01

    Cryptosporidium is one of the most common zoonotic waterborne parasitic diseases worldwide and represents a major public health concern of water utilities in developed nations. As animals in catchments can shed human-infectious Cryptosporidium oocysts, determining the potential role of animals in dissemination of zoonotic Cryptosporidium to drinking water sources is crucial. In the present study, a total of 952 animal faecal samples from four dominant species (kangaroos, rabbits, cattle and sheep) inhabiting Sydney’s drinking water catchments were screened for the presence of Cryptosporidium using a quantitative PCR (qPCR) and positives sequenced at multiple loci. Cryptosporidium species were detected in 3.6% (21/576) of kangaroos, 7.0% (10/142) of cattle, 2.3% (3/128) of sheep and 13.2% (14/106) of rabbit samples screened. Sequence analysis of a region of the 18S rRNA locus identified C. macropodum and C. hominis in 4 and 17 isolates from kangaroos respectively, C. hominis and C. parvum in 6 and 4 isolates respectively each from cattle, C. ubiquitum in 3 isolates from sheep and C. cuniculus in 14 isolates from rabbits. All the Cryptosporidium species identified were zoonotic species with the exception of C. macropodum. Subtyping using the 5’ half of gp60 identified C. hominis IbA10G2 (n = 12) and IdA15G1 (n = 2) in kangaroo faecal samples; C. hominis IbA10G2 (n = 4) and C. parvum IIaA18G3R1 (n = 4) in cattle faecal samples, C. ubiquitum subtype XIIa (n = 1) in sheep and C. cuniculus VbA23 (n = 9) in rabbits. Additional analysis of a subset of samples using primers targeting conserved regions of the MIC1 gene and the 3’ end of gp60 suggests that the C. hominis detected in these animals represent substantial variants that failed to amplify as expected. The significance of this finding requires further investigation but might be reflective of the ability of this C. hominis variant to infect animals. The finding of zoonotic Cryptosporidium species in these

  13. Application of oligonucleotide microarrays for bacterial source tracking of environmental Enterococcus sp. isolates.

    PubMed

    Indest, Karl J; Betts, Kelley; Furey, John S

    2005-04-01

    In an effort towards adapting new and defensible methods for assessing and managing the risk posed by microbial pollution, we evaluated the utility of oligonucleotide microarrays for bacterial source tracking (BST) of environmental Enterococcus sp. isolates derived from various host sources. Current bacterial source tracking approaches rely on various phenotypic and genotypic methods to identify sources of bacterial contamination resulting from point or non-point pollution. For this study Enterococcus sp. isolates originating from deer, bovine, gull, and human sources were examined using microarrays. Isolates were subjected to Box PCR amplification and the resulting amplification products labeled with Cy5. Fluorescent-labeled templates were hybridized to in-house constructed nonamer oligonucleotide microarrays consisting of 198 probes. Microarray hybridization profiles were obtained using the ArrayPro image analysis software. Principal Components Analysis (PCA) and Hierarchical Cluster Analysis (HCA) were compared for their ability to visually cluster microarray hybridization profiles based on the environmental source from which the Enterococcus sp. isolates originated. The PCA was visually superior at separating origin-specific clusters, even for as few as 3 factors. A Soft Independent Modeling (SIM) classification confirmed the PCA, resulting in zero misclassifications using 5 factors for each class. The implication of these results for the application of random oligonucleotide microarrays for BST is that, given the reproducibility issues, factor-based variable selection such as in PCA and SIM greatly outperforms dendrogram-based similarity measures such as in HCA and K-Nearest Neighbor KNN.

  14. Application of Oligonucleotide Microarrays for Bacterial Source Tracking of Environmental Enterococcus sp. Isolates

    PubMed Central

    Indest, Karl J.; Betts, Kelley; Furey, John S.

    2005-01-01

    In an effort towards adapting new and defensible methods for assessing and managing the risk posed by microbial pollution, we evaluated the utility of oligonucleotide microarrays for bacterial source tracking (BST) of environmental Enterococcus sp. isolates derived from various host sources. Current bacterial source tracking approaches rely on various phenotypic and genotypic methods to identify sources of bacterial contamination resulting from point or non-point pollution. For this study Enterococcus sp. isolates originating from deer, bovine, gull, and human sources were examined using microarrays. Isolates were subjected to Box PCR amplification and the resulting amplification products labeled with Cy5. Fluorescent-labeled templates were hybridized to in-house constructed nonamer oligonucleotide microarrays consisting of 198 probes. Microarray hybridization profiles were obtained using the ArrayPro image analysis software. Principal Components Analysis (PCA) and Hierarchical Cluster Analysis (HCA) were compared for their ability to visually cluster microarray hybridization profiles based on the environmental source from which the Enterococcus sp. isolates originated. The PCA was visually superior at separating origin-specific clusters, even for as few as 3 factors. A Soft Independent Modeling (SIM) classification confirmed the PCA, resulting in zero misclassifications using 5 factors for each class. The implication of these results for the application of random oligonucleotide microarrays for BST is that, given the reproducibility issues, factor-based variable selection such as in PCA and SIM greatly outperforms dendrogram-based similarity measures such as in HCA and K-Nearest Neighbor KNN. PMID:16705816

  15. CONCENTRATION AND DETECTION OF CRYPTOSPORIDIUM OOCYSTS IN SURFACE WATER SAMPLES BY METHOD 1622 USING ULTRAFILTRATION AND CAPSULE FILTRATION

    EPA Science Inventory

    The protozoan parasite cryptosporidium parvum is known to occur widely in both source and drinking water and has caused waterborne outbreaks of gastroenteritis. To improve monitoring, the USEPA developed Method 1622 for isolation and detection of cryptosporidim oocysts in water. ...

  16. Biology of Cryptosporidium from marsupial hosts.

    PubMed

    Power, Michelle L

    2010-01-01

    The majority of biological data on Cryptosporidium has been collected from humans and domestic animal hosts which creates a bias in knowledge on the biodiversity and evolution of this parasite genus. Further to understanding Cryptosporidium biology are studies encompassing broad hosts that represent diverse taxa sampled across wide geographic ranges. Marsupials represent a group of wildlife hosts from which limited information on Cryptosporidium is available. As marsupial hosts are an ancient mammalian lineage they represent an important group for studying parasite evolution. This review summarises information of the biology, epidemiology and evolution of Cryptosporidium in marsupial hosts, and discusses the importance of further understanding interactions in this parasite-host system.

  17. Enterococcus and Escherichia coli fecal source apportionment with microbial source tracking genetic markers - is it feasible?

    EPA Science Inventory

    Fecal pollution is measured in surface waters using culture-based measurements of enterococci and Escherichia coli bacteria. Source apportionment of these two fecal indicator bacteria is an urgent need for prioritizing remediation efforts and quantifying health risks associated...

  18. Source tracking using microbial community fingerprints: Method comparison with hydrodynamic modelling.

    PubMed

    McCarthy, D T; Jovanovic, D; Lintern, A; Teakle, I; Barnes, M; Deletic, A; Coleman, R; Rooney, G; Prosser, T; Coutts, S; Hipsey, M R; Bruce, L C; Henry, R

    2017-02-01

    Urban estuaries around the world are experiencing contamination from diffuse and point sources, which increases risks to public health. To mitigate and manage risks posed by elevated levels of contamination in urban waterways, it is critical to identify the primary water sources of contamination within catchments. Source tracking using microbial community fingerprints is one tool that can be used to identify sources. However, results derived from this approach have not yet been evaluated using independent datasets. As such, the key objectives of this investigation were: (1) to identify the major sources of water responsible for bacterial loadings within an urban estuary using microbial source tracking (MST) using microbial communities; and (2) to evaluate this method using a 3-dimensional hydrodynamic model. The Yarra River estuary, which flows through the city of Melbourne in South-East Australia was the focus of this study. We found that the water sources contributing to the bacterial community in the Yarra River estuary varied temporally depending on the estuary's hydrodynamic conditions. The water source apportionment determined using microbial community MST correlated to those determined using a 3-dimensional hydrodynamic model of the transport and mixing of a tracer in the estuary. While there were some discrepancies between the two methods, this investigation demonstrated that MST using bacterial community fingerprints can identify the primary water sources of microorganisms in an estuarine environment. As such, with further optimization and improvements, microbial community MST has the potential to become a powerful tool that could be practically applied in the mitigation of contaminated aquatic systems.

  19. Human-, Ovine-, and Bovine-Specific Viral Source Tracking Tools to Discriminate Between the Major Fecal Sources in Agricultural Waters.

    PubMed

    Rusiñol, Marta; Moriarty, Elaine; Lin, Susan; Bofill-Mas, Sílvia; Gilpin, Brent

    2016-03-01

    This study evaluated the sources of fecal contamination in different river catchments, using a combination of microbial source tracking tools, for human, ruminant, ovine and bovine livestock, in order to define appropriate water management strategies. Every source of waterway pollution was evaluated in river water samples from one urban river catchment and two important farming regions in New Zealand. Fecal pollution was initially measured by testing Escherichia coli and evaluating the presence of human- and ruminant-associated DNA markers of Bacteroidales (BiAdo, BacHum-UCD, BacH, and BacR) and human and ruminant fecal sterols/stanols ratios. Then specific fecal pollution sources were assessed with previously reported quantitative PCR assays targeting human-, bovine-, and ovine-specific viruses: human adenoviruses (HAdV), human JC polyomaviruses, bovine polyomaviruses (BPyV), and ovine polyomaviruses (OPyV). High level of ruminant fecal contamination was detected all over the farming areas, whereas no ruminant sources were identified in the urban river sampling sites. BacR was the most frequently observed ruminant marker and OPyV and BPyV allowed the identification of ovine and bovine fecal sources. The human fecal viral marker (HAdV) was the most frequently observed human marker, highly abundant in the urban sites, and also present in farming areas. This is the first study using simultaneously the ovine and the bovine viral markers to identify and quantify both bovine and ovine fecal pollution.

  20. Biology, persistence and detection of Cryptosporidium parvum and Cryptosporidium hominis oocyst.

    PubMed

    Carey, C M; Lee, H; Trevors, J T

    2004-02-01

    Cryptosporidium parvum and Cryptosporidium hominis are obligate enteric protozoan parasites which infect the gastrointestinal tract of animals and humans. The mechanism(s) by which these parasites cause gastrointestinal distress in their hosts is not well understood. The risk of waterborne transmission of Cryptosporidium is a serious global issue in drinking water safety. Oocysts from these organisms are extremely robust, prevalent in source water supplies and capable of surviving in the environment for extended periods of time. Resistance to conventional water treatment by chlorination, lack of correlation with biological indicator microorganisms and the absence of adequate methods to detect the presence of infectious oocysts necessitates the development of consistent and effective means of parasite removal from the water supply. Additional research into improving water treatment and sewage treatment practices is needed, particularly in testing the efficiency of ozone in oocyst inactivation. Timely and efficient detection of infectious C. parvum and C. hominis oocysts in environmental samples requires the development of rapid and sensitive techniques for the concentration, purification and detection of these parasites. A major factor confounding proper detection remains the inability to adequately and efficiently concentrate oocysts from environmental samples, while limiting the presence of extraneous materials. Molecular-based techniques are the most promising methods for the sensitive and accurate detection of C. parvum and C. hominis. With the availability of numerous target sequences, RT-PCR will likely emerge as an important method to assess oocyst viability. In addition, a multiplex PCR for the simultaneous detection of C. parvum, C. hominis and other waterborne pathogens such as Giardia lamblia would greatly benefit the water industry and protect human health.

  1. Watershed Assessment with Beach Microbial Source Tracking and Outcomes of Resulting Gull Management.

    PubMed

    Goodwin, Kelly D; Gruber, Steve; Vondrak, Mary; Crumpacker, Andrea

    2016-09-20

    Total maximum daily load (TMDL) implementation at a southern California beach involved ultraviolet treatment of watershed drainage that provided >97% reduction in fecal indicator bacteria (FIB) concentrations. However, this pollutant control measure did not provide sufficient improvement of beach water quality, prompting further assessment. Investigation included microbial source tracking (MST) for human, gull, and canine fecal sources, monitoring of enterococci and fecal coliform, and measurement of chemical and physical water quality parameters for samples collected from watershed, groundwater, and beach sites, including a beach scour pond and tidal creek. FIB variability remained poorly modeled in regression analysis. However, MST revealed correlations between FIB and gull source tracking markers, leading to recommendations to manage gulls as a pollutant source. Beach conditions were followed for three years after implementation of a best management practice (BMP) to abate gulls using a falconry program for the beach and an upland landfill. The gull abatement BMP was associated with improved beach water quality, and this appears to be the first report of falconry in the context of TMDL implementation. Overall, MST data enabled management action despite an inability to fully model FIB dynamics in the coupled watershed-beach system.

  2. Microbial source tracking in impaired watersheds using PhyloChip and machine-learning classification.

    PubMed

    Dubinsky, Eric A; Butkus, Steven R; Andersen, Gary L

    2016-11-15

    Sources of fecal indicator bacteria are difficult to identify in watersheds that are impacted by a variety of non-point sources. We developed a molecular source tracking test using the PhyloChip microarray that detects and distinguishes fecal bacteria from humans, birds, ruminants, horses, pigs and dogs with a single test. The multiplexed assay targets 9001 different 25-mer fragments of 16S rRNA genes that are common to the bacterial community of each source type. Both random forests and SourceTracker were tested as discrimination tools, with SourceTracker classification producing superior specificity and sensitivity for all source types. Validation with 12 different mammalian sources in mixtures found 100% correct identification of the dominant source and 84-100% specificity. The test was applied to identify sources of fecal indicator bacteria in the Russian River watershed in California. We found widespread contamination by human sources during the wet season proximal to settlements with antiquated septic infrastructure and during the dry season at beaches during intense recreational activity. The test was more sensitive than common fecal indicator tests that failed to identify potential risks at these sites. Conversely, upstream beaches and numerous creeks with less reliance on onsite wastewater treatment contained no fecal signal from humans or other animals; however these waters did contain high counts of fecal indicator bacteria after rain. Microbial community analysis revealed that increased E. coli and enterococci at these locations did not co-occur with common fecal bacteria, but rather co-varied with copiotrophic bacteria that are common in freshwaters with high nutrient and carbon loading, suggesting runoff likely promoted the growth of environmental strains of E. coli and enterococci. These results indicate that machine-learning classification of PhyloChip microarray data can outperform conventional single marker tests that are used to assess health

  3. Adaptive Environmental Source Localization and Tracking with Unknown Permittivity and Path Loss Coefficients †

    PubMed Central

    Fidan, Barış; Umay, Ilknur

    2015-01-01

    Accurate signal-source and signal-reflector target localization tasks via mobile sensory units and wireless sensor networks (WSNs), including those for environmental monitoring via sensory UAVs, require precise knowledge of specific signal propagation properties of the environment, which are permittivity and path loss coefficients for the electromagnetic signal case. Thus, accurate estimation of these coefficients has significant importance for the accuracy of location estimates. In this paper, we propose a geometric cooperative technique to instantaneously estimate such coefficients, with details provided for received signal strength (RSS) and time-of-flight (TOF)-based range sensors. The proposed technique is integrated to a recursive least squares (RLS)-based adaptive localization scheme and an adaptive motion control law, to construct adaptive target localization and adaptive target tracking algorithms, respectively, that are robust to uncertainties in aforementioned environmental signal propagation coefficients. The efficiency of the proposed adaptive localization and tracking techniques are both mathematically analysed and verified via simulation experiments. PMID:26690441

  4. COMPARATIVE DIVERSITY OF FECAL BACTERIA IN AGRICULTURALLY SIGNIFICANT ANIMALS TO IDENTIFY ALTERNATIVE TARGETS FOR MICROBIAL SOURCE TRACKING

    EPA Science Inventory

    Animals of agricultural significance contribute a large percentage of fecal pollution to waterways via runoff contamination. The premise of microbial source tracking is to utilize fecal bacteria to identify target populations which are directly correlated to specific animal feces...

  5. Use of Enterococcus, BST and sterols as indicators for poultry pollution source tracking in surface and groundwater

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study has applied Enterococcus, Bacterial Source Tracking (BST) and sterol analysis for pollution source identification from poultry sources. Fecal contamination was detected in 100% of surface water and 15% of groundwater sites tested. E. faecium was the dominant species in aged litter sampl...

  6. Genetic Diversity of Cryptosporidium spp. in Captive Reptiles

    PubMed Central

    Xiao, Lihua; Ryan, Una M.; Graczyk, Thaddeus K.; Limor, Josef; Li, Lixia; Kombert, Mark; Junge, Randy; Sulaiman, Irshad M.; Zhou, Ling; Arrowood, Michael J.; Koudela, Břetislav; Modrý, David; Lal, Altaf A.

    2004-01-01

    The genetic diversity of Cryptosporidium in reptiles was analyzed by PCR-restriction fragment length polymorphism and sequence analysis of the small subunit rRNA gene. A total of 123 samples were analyzed, of which 48 snake samples, 24 lizard samples, and 3 tortoise samples were positive for Cryptosporidium. Nine different types of Cryptosporidium were found, including Cryptosporidium serpentis, Cryptosporidium desert monitor genotype, Cryptosporidium muris, Cryptosporidium parvum bovine and mouse genotypes, one C. serpentis-like parasite in a lizard, two new Cryptosporidium spp. in snakes, and one new Cryptosporidium sp. in tortoises. C. serpentis and the desert monitor genotype were the most common parasites and were found in both snakes and lizards, whereas the C. muris and C. parvum parasites detected were probably the result of ingestion of infected rodents. Sequence and biologic characterizations indicated that the desert monitor genotype was Cryptosporidium saurophilum. Two host-adapted C. serpentis genotypes were found in snakes and lizards. PMID:14766569

  7. Genetic diversity of Cryptosporidium spp. in captive reptiles.

    PubMed

    Xiao, Lihua; Ryan, Una M; Graczyk, Thaddeus K; Limor, Josef; Li, Lixia; Kombert, Mark; Junge, Randy; Sulaiman, Irshad M; Zhou, Ling; Arrowood, Michael J; Koudela, Bretislav; Modrý, David; Lal, Altaf A

    2004-02-01

    The genetic diversity of Cryptosporidium in reptiles was analyzed by PCR-restriction fragment length polymorphism and sequence analysis of the small subunit rRNA gene. A total of 123 samples were analyzed, of which 48 snake samples, 24 lizard samples, and 3 tortoise samples were positive for Cryptosporidium: Nine different types of Cryptosporidium were found, including Cryptosporidium serpentis, Cryptosporidium desert monitor genotype, Cryptosporidium muris, Cryptosporidium parvum bovine and mouse genotypes, one C. serpentis-like parasite in a lizard, two new Cryptosporidium spp. in snakes, and one new Cryptosporidium sp. in tortoises. C. serpentis and the desert monitor genotype were the most common parasites and were found in both snakes and lizards, whereas the C. muris and C. parvum parasites detected were probably the result of ingestion of infected rodents. Sequence and biologic characterizations indicated that the desert monitor genotype was Cryptosporidium saurophilum. Two host-adapted C. serpentis genotypes were found in snakes and lizards.

  8. Modelling Cryptosporidium oocysts transport in small ungauged agricultural catchments.

    PubMed

    Tang, Jialiang; McDonald, Stephen; Peng, Xinhua; Samadder, Sukha R; Murphy, Thomas M; Holden, Nicholas M

    2011-06-01

    Cryptosporidium is an environmentally robust pathogen that has caused severe waterborne disease outbreaks worldwide. The main source of zoonotic Cryptosporidium parvum oocysts in human drinking water is likely to be from farm animals via catchment pathways with water as the main transport vector. The vast majority of small agricultural catchments are ungauged therefore it is difficult to use a process model to predict and understand the mechanisms and activities that regulate the risk of surface water contamination from agricultural areas. For this study, two ungauged agricultural catchments in Ireland were used to model Cryptosporidium oocyst transport using SWAT2005 on a daily basis with reference data from adjacent catchment gauging stations. The results indicated that SWAT2005 could simulate stream flow with good agreement between prediction and observation on a monthly basis (R(2) from 0.94 to 0.83 and E (efficiency) from 0.92 to 0.66), but Cryptosporidium oocyst concentration results were less reliable (R(2) from 0.20 to 0.37, P < 0.05; with poor E -0.37 to -2.57). A sensitivity analysis using independent parameter perturbation indicated that temperature was the most important parameter regulating oocyst transport in the study catchments and that the timing of manure application relative to the occurrence of water runoff event was critical. The results also showed that grazing management had little influence on predicted oocyst transport while fields fertilized with manure were the key critical source areas for microbial contaminations in the study catchments. It was concluded that the approach presented could be used to assist with understanding the epidemiology of waterborne cryptosporidiosis outbreaks and to improve catchment management for the safety of the general public health.

  9. A perspective on Cryptosporidium and Giardia, with an emphasis on bovines and recent epidemiological findings.

    PubMed

    Abeywardena, Harshanie; Jex, Aaron R; Gasser, Robin B

    2015-04-01

    Cryptosporidium and Giardia are two common aetiological agents of infectious enteritis in humans and animals worldwide. These parasitic protists are usually transmitted by the faecal-oral route, following the ingestion of infective stages (oocysts or cysts). An essential component of the control of these parasitic infections, from a public health perspective, is an understanding of the sources and routes of transmission in different geographical regions. Bovines are considered potential sources of infection for humans, because species and genotypes of Cryptosporidium and Giardia infecting humans have also been isolated from cattle in molecular parasitological studies. However, species and genotypes of Cryptosporidium and Giardia of bovids, and the extent of zoonotic transmission in different geographical regions in the world, are still relatively poorly understood. The purpose of this article is to (1) provide a brief background on Cryptosporidium and Giardia, (2) review some key aspects of the molecular epidemiology of cryptosporidiosis and giardiasis in animals, with an emphasis on bovines, (3) summarize research of Cryptosporidium and Giardia from cattle and water buffaloes in parts of Australasia and Sri Lanka, considering public health aspects and (4) provide a perspective on future avenues of study. Recent studies reinforce that bovines harbour Cryptosporidium and Giardia that likely pose a human health risk and highlight the need for future investigations of the biology, population genetics and transmission dynamics of Cryptosporidium and Giardia in cattle, water buffaloes and other ruminants in different geographical regions, the fate and transport of infective stages following their release into the environment, as well as for improved strategies for the control and prevention of cryptosporidiosis and giardiasis, guided by molecular epidemiological studies.

  10. Classification tree method for bacterial source tracking with antibiotic resistance analysis data.

    PubMed

    Price, Bertram; Venso, Elichia A; Frana, Mark F; Greenberg, Joshua; Ware, Adam; Currey, Lee

    2006-05-01

    Various statistical classification methods, including discriminant analysis, logistic regression, and cluster analysis, have been used with antibiotic resistance analysis (ARA) data to construct models for bacterial source tracking (BST). We applied the statistical method known as classification trees to build a model for BST for the Anacostia Watershed in Maryland. Classification trees have more flexibility than other statistical classification approaches based on standard statistical methods to accommodate complex interactions among ARA variables. This article describes the use of classification trees for BST and includes discussion of its principal parameters and features. Anacostia Watershed ARA data are used to illustrate the application of classification trees, and we report the BST results for the watershed.

  11. Molecular epidemiology of Cryptosporidium in livestock animals and humans in the Ismailia province of Egypt.

    PubMed

    Helmy, Yosra A; Krücken, Jürgen; Nöckler, Karsten; von Samson-Himmelstjerna, Georg; Zessin, Karl-H

    2013-03-31

    The zoonotic potential of Cryptosporidium was studied in one of the most densely populated provinces of Egypt regarding livestock and people. In a representative survey, faecal samples from cattle, buffalo and stool samples from diarrhoeic children (<10 years) were investigated. Parameters assumed to be related to cryptosporidiosis were recorded for animals and children. Animal samples (804) were examined by the Copro-antigen RIDA(®)QUICK test, followed by PCRs targeting the 18S rDNA and gp60 genes for antigen-positive and 10% randomly selected negative samples. All 165 human samples were tested by both methods. The overall estimated prevalence of Cryptosporidium in ruminants was 32.2%, without significant difference between animal species. PCR identified 65.7% Cryptosporidium parvum, 11.8% Cryptosporidium ryanae, 4.1% Cryptosporidium bovis, and combinations of C. parvum plus C. ryanae (11.2%), C. parvum plus C. bovis (5.3%) and of C. parvum plus Cryptosporidium andersoni (1.8%), also without significant differences in species occurrence between cattle and buffalos. The human Cryptosporidium spp. prevalence was 49.1%, of which 60.5% were Cryptosporidium hominis, 38.2% C. parvum and 1.2% C. parvum plus C. bovis. Analysis of gp60 variants allocated C. parvum found in animals to the zoonotic subtype family IIa (18.9%, subtype IIaA15G1R1 only) and to IId (81.1%, mostly IIdA20G1). In humans 50% were classified as subtype family IIa (IIaA15G1R1 and IIaA15G2R1) and 50% were IIdA20G1. C. andersoni occurred only in cattle older than 1 year. In contrast, mono-infections with one of the three single Cryptosporidium species and the three combinations with C. parvum were more prevalent in cattle and buffaloes younger than 1 year, particularly in those younger than 3 months, and were predominantly subtype family IId. In human samples no Cryptosporidium were identified in children younger than 7 months. Neither place of residence nor the source of drinking-water had measurable

  12. WATER FROM DIFFERENT SOURCES USED FOR THE IRRIGATION OF VEGETABLES TO BE MARKETED: RESEARCH ON Cryptosporidium spp., Giardia spp., AND COLIFORMS IN PARANA, BRAZIL

    PubMed Central

    TIYO, Rogerio; de SOUZA, Carla Zangari; NISHI, Letícia; BRUSTOLIN, Camila Fernanda; RATTI, Bianca Altrão; FALAVIGNA GUILHERME, Ana Lucia

    2015-01-01

    SUMMARY The aim of this work was to compare, from a parasitological ( Cryptosporidiumspp. and Giardia duodenalis), bacteriological (total and thermotolerants coliforms) and physicochemical perspective, water sources used for drinking and irrigation of vegetables intended to be sold for human consumption. From January 2010 to May 2011, samples of different water sources from vegetable producing properties were collected; 100 liters for parasitological analysis, 200 mL for bacteriological analysis, and five liters for physicochemical analysis. Water samples were filtered under vacuum with a kit containing a cellulose acetate membrane filter, 1.2 µm (Millipore(r), Barueri, SP, Brazil). The material retained on the membrane was mechanically extracted and analyzed by direct immunofluorescence (Merifluor(r)kit). From 20 rural properties investigated, 10 had artesian wells (40 samples), 10 had common wells (40 samples), and one had a mine (four samples), the latter contaminated by Cryptosporidiumspp. In samples from artesian wells, 90 to 130 meters depth, 42.5% were positive for total coliforms and 5.0% were identified to have abnormal coloration. From the samples of common wells, 14 to 37 meters depth, 87.5% were contaminated with total coliforms, 82.5% were positive for thermotolerant coliforms, and 12.5% had color abnormalities. We did not detect the presence of Giardiaspp. or Cryptosporidiumspp. in artesian and common wells. The use of artesian or common wells is an important step in the control of the spreading of zoonoses, particularly Cryptosporidiumspp. and Giardiaspp., as well as artesian wells for coliform control in local production of vegetables to be marketed. PMID:26422158

  13. Identification and tracking of harmonic sources in a power system using a Kalman filter

    SciTech Connect

    Ma, H.; Girgis, A.A.

    1996-07-01

    In this paper, two problems have been addressed on harmonic sources identification: the optimal locations of a limited number of harmonic meters and the optimal dynamic estimates of harmonic source locations and their injections in unbalanced three-phase power systems. A Kalman filtering is used to attack these problems. System error covariance analysis by the Kalman filter associated with a harmonic injection estimate determines the optimal arrangement of limited harmonic meters. Based on the optimally-arranged harmonic metering locations, the Kalman filter then yields the optimal dynamic estimates of harmonic injections with a few noisy harmonic measurements. The method is dynamic and has the capability of identifying, analyzing and tracking each harmonic injection at all buses in unbalanced three-phase power systems. Actual recorded harmonic measurements and simulated data in a power distribution system are provided to prove the efficiency of this approach.

  14. Microbial source tracking markers at three inland recreational lakes in Ohio, 2011

    USGS Publications Warehouse

    Francy, Donna S.; Stelzer, Erin A.

    2012-01-01

    During the 2011 recreational season, samples were collected for E. coli and microbial source tracking (MST) marker concentrations to begin to understand potential sources of fecal contamination at three inland recreational lakes in Ohio - Buckeye, Atwood, and Tappan Lakes. The results from 32 regular samples, 4 field blanks, and 7 field replicates collected at 5 sites are presented in this report. At the three lakes, the ruminant-associated marker was found most often (57-73 percent of samples) but at estimated quantities, followed by the dog-associated marker (30-43 percent of samples). The human-associated marker was found in 14 and 50 percent of samples from Atwood and Tappan Lakes, respectively, but was not found in any samples from the two Buckeye Lake sites. The gull-associated marker was detected in only two samples, both from Tappan Lake.

  15. Sensivitity improvement in low-profile distributed detector systems for tracking sources in transit.

    SciTech Connect

    Vilim, R. B.; Klann, R.; Campos, C.; Medley, T.; Nuclear Engineering Division

    2007-01-01

    The RadTrac real-time detection and tracking software runs on a laptop computer networked to gamma-radiation detectors. A probabilistic estimate for source position is generated by combining measured count rate data with a first-principles stochastic model for the space and time dependence of count rates and knowledge of detector intrinsic efficiency. Recent development work has focused on improving RadTrac sensitivity in low-count rate situations. A method has been developed for processing count rates by energy according to that part of the energy spectrum with the greatest signal-to-noise ratio. In addition a method has been developed that places constraints on the solution that are physically appropriate when count rates approach background. In both instances experiments with a weak source confirmed the uncertainty in estimated position is reduced.

  16. Source tracking swine fecal waste in surface water proximal to swine concentrated animal feeding operations.

    PubMed

    Heaney, Christopher D; Myers, Kevin; Wing, Steve; Hall, Devon; Baron, Dothula; Stewart, Jill R

    2015-04-01

    Swine farming has gone through many changes in the last few decades, resulting in operations with a high animal density known as confined animal feeding operations (CAFOs). These operations produce a large quantity of fecal waste whose environmental impacts are not well understood. The purpose of this study was to investigate microbial water quality in surface waters proximal to swine CAFOs including microbial source tracking of fecal microbes specific to swine. For one year, surface water samples at up- and downstream sites proximal to swine CAFO lagoon waste land application sites were tested for fecal indicator bacteria (fecal coliforms, Escherichia coli and Enterococcus) and candidate swine-specific microbial source-tracking (MST) markers (Bacteroidales Pig-1-Bac, Pig-2-Bac, and Pig-Bac-2, and methanogen P23-2). Testing of 187 samples showed high fecal indicator bacteria concentrations at both up- and downstream sites. Overall, 40%, 23%, and 61% of samples exceeded state and federal recreational water quality guidelines for fecal coliforms, E. coli, and Enterococcus, respectively. Pig-1-Bac and Pig-2-Bac showed the highest specificity to swine fecal wastes and were 2.47 (95% confidence interval [CI]=1.03, 5.94) and 2.30 times (95% CI=0.90, 5.88) as prevalent proximal down- than proximal upstream of swine CAFOs, respectively. Pig-1-Bac and Pig-2-Bac were also 2.87 (95% CI=1.21, 6.80) and 3.36 (95% CI=1.34, 8.41) times as prevalent when 48 hour antecedent rainfall was greater than versus less than the mean, respectively. Results suggest diffuse and overall poor sanitary quality of surface waters where swine CAFO density is high. Pig-1-Bac and Pig-2-Bac are useful for tracking off-site conveyance of swine fecal wastes into surface waters proximal to and downstream of swine CAFOs and during rain events.

  17. Source tracking swine fecal waste in surface water proximal to swine concentrated animal feeding operations

    PubMed Central

    Heaney, Christopher D.; Myers, Kevin; Wing, Steve; Hall, Devon; Baron, Dothula; Stewart, Jill R.

    2015-01-01

    Swine farming has gone through many changes in the last few decades, resulting in operations with a high animal density known as confined animal feeding operations (CAFOs). These operations produce a large quantity of fecal waste whose environmental impacts are not well understood. The purpose of this study was to investigate microbial water quality in surface waters proximal to swine CAFOs including microbial source tracking of fecal microbes specific to swine. For one year, surface water samples at up- and downstream sites proximal to swine CAFO lagoon waste land application sites were tested for fecal indicator bacteria (fecal coliforms, Escherichia coli and Enterococcus) and candidate swine-specific microbial source-tracking (MST) markers (Bacteroidales Pig-1-Bac, Pig-2-Bac, and Pig-Bac-2, and methanogen P23-2). Testing of 187 samples showed high fecal indicator bacteria concentrations at both up- and downstream sites. Overall, 40%, 23%, and 61% of samples exceeded state and federal recreational water quality guidelines for fecal coliforms, E. coli, and Enterococcus, respectively. Pig-1-Bac and Pig-2-Bac showed the highest specificity to swine fecal wastes and were 2.47 (95% confidence interval [CI] = 1.03, 5.94) and 2.30 times (95% CI = 0.90, 5.88) as prevalent proximal down- than proximal upstream of swine CAFOs, respectively. Pig-1-Bac and Pig-2-Bac were also 2.87 (95% CI = 1.21, 6.80) and 3.36 (95% CI = 1.34, 8.41) times as prevalent when 48 hour antecedent rainfall was greater than versus less than the mean, respectively. Results suggest diffuse and overall poor sanitary quality of surface waters where swine CAFO density is high. Pig-1-Bac and Pig-2-Bac are useful for tracking off-site conveyance of swine fecal wastes into surface waters proximal to and downstream of swine CAFOs and during rain events. PMID:25600418

  18. Radiological source tracking in oil/gas, medical and other industries: requirements and specifications for passive RFID technology

    SciTech Connect

    Dowla, Farid U.

    2016-01-01

    Subsurface sensors that employ radioisotopes, such 241Am-Be and 137Cs, for reservoir characterization must be tracked for safety and security reasons. Other radiological sources are also widely used in medicine. The radiological source containers, in both applications, are small, mobile and used widely worldwide. The nuclear sources pose radiological dispersal device (RDD) security risks. Security concerns with the industrial use of radionuclide sources is in fact quite high as it is estimated that each year hundreds of sealed sources go missing, either lost or stolen. Risk mitigation efforts include enhanced regulations, source-use guidelines, research and development on electronic tracking of sources. This report summarizes the major elements of the requirements and operational concepts of nuclear sources with the goal of developing automated electronic tagging and locating systems.

  19. Cryptosporidium parvum is not transmissible to fish, amphibians, or reptiles.

    PubMed

    Graczyk, T K; Fayer, R; Cranfield, M R

    1996-10-01

    A recent report suggested that an isolate of Cryptosporidium parvum had established infections in fish, amphibians, and reptiles and raises concern that animals other than mammals might be a potential source of waterborne Cryptosporidium oocysts. To test this possibility, viable C. parvum oocysts, infectious for neonatal BALB/c mice, were delivered by gastric intubation to bluegill sunfish, poison-dart frogs, African clawed frogs, bearded dragon lizards, and corn snakes. Histological sections of the stomach, jejunum, ileum, and cloaca prepared from tissues collected on days 7 and 14 postinoculation (PI) were negative for Cryptosporidium developmental stages. However, inoculum-derived oocysts were detectable by fluorescein-labeled monoclonal antibody in feces of inoculated animals from day 1 to day 12 PI in fish and frogs, and up to day 14 PI in lizards. Snakes did not defecate for 14 days PI. Impression smears taken at necropsy on days 7 and 14 PI revealed C. parvum oocysts in the lumen of the cloaca of 2 fish and 1 lizard on day 7 PI only. Because tissue stages of the pathogen were not found, it appears that C. parvum was not heterologously transmitted to lower vertebrates. Under certain circumstances, however, such as after the ingestion of C. parvum-infected prey, lower vertebrates may disseminate C. parvum oocysts in the environment.

  20. Chemical Source Tracking of Bacterial Contamination Using Micropollutants - A Karst Aquifer Case Study

    NASA Astrophysics Data System (ADS)

    Zirlewagen, Johannes; Hillebrand, Olav; Nödler, Karsten; Licha, Tobias; Schiperski, Ferry; Stange, Claudia; Tiehm, Andreas; Scheytt, Traugott

    2015-04-01

    Karst aquifers are important drinking water resources in many parts of the world, though they are well known for their high vulnerability to contamination. Rainfall and snowmelt often trigger temporary contamination of karst water resources. Free-range animal breeding and application of manure on the one hand and sewage leakage or spillage on the other hand are usually regarded as main sources for fecal contamination. But distinction of their respective contributions is difficult. This study investigates the feasibility to track the origin of fecal contamination from the occurrences of indicator bacteria and chemical source indicators in karst spring water. The study site is the 45 km² rural catchment of the perennial karst spring Gallusquelle in SW-Germany (mean discharge: 0.5 m³/s). Overflow events of a stormwater detention basin (combined sewer system) are known to impact water quality at the spring. There is no free-range animal breeding in the catchment but intense application of manure. Following two heavy rainfall events with overflow of the stormwater detention basin, spring water was sampled over several days. Samples were analysed for indicator bacteria (total Coliform, E. coli, Enterococci) and 57 micropollutants, among them cyclamate and metazachlor. For the Gallusquelle catchment the artificial sweetener cyclamate and the herbicide metazachlor have been established as source specific indicators, the former for the sewer system and the latter for cropland. Though recharge in the Gallusquelle catchment is predominantly diffuse, there is a significant portion of direct recharge reflected by distinct breakthrough curves for cyclamate and metazachlor. The breakthrough of indicator bacteria coincides very well with the occurrence of both, cyclamate and metazachlor. However, indicator bacteria cannot be unambiguously tracked back to a specific source.

  1. Data Fusion for a Vision-Radiological System for Source Tracking and Discovery

    SciTech Connect

    Enqvist, Andreas; Koppal, Sanjeev

    2015-07-01

    A multidisciplinary approach to allow the tracking of the movement of radioactive sources by fusing data from multiple radiological and visual sensors is under development. The goal is to improve the ability to detect, locate, track and identify nuclear/radiological threats. The key concept is that such widely available visual and depth sensors can impact radiological detection, since the intensity fall-off in the count rate can be correlated to movement in three dimensions. To enable this, we pose an important question; what is the right combination of sensing modalities and vision algorithms that can best compliment a radiological sensor, for the purpose of detection and tracking of radioactive material? Similarly what is the best radiation detection methods and unfolding algorithms suited for data fusion with tracking data? Data fusion of multi-sensor data for radiation detection have seen some interesting developments lately. Significant examples include intelligent radiation sensor systems (IRSS), which are based on larger numbers of distributed similar or identical radiation sensors coupled with position data for network capable to detect and locate radiation source. Other developments are gamma-ray imaging systems based on Compton scatter in segmented detector arrays. Similar developments using coded apertures or scatter cameras for neutrons have recently occurred. The main limitation of such systems is not so much in their capability but rather in their complexity and cost which is prohibitive for large scale deployment. Presented here is a fusion system based on simple, low-cost computer vision and radiological sensors for tracking of multiple objects and identifying potential radiological materials being transported or shipped. The main focus of this work is the development on two separate calibration algorithms for characterizing the fused sensor system. The deviation from a simple inverse square-root fall-off of radiation intensity is explored and

  2. First molecular characterization of Cryptosporidium in Yemen.

    PubMed

    Alyousefi, N A; Mahdy, M A K; Lim, Y A L; Xiao, L; Mahmud, R

    2013-05-01

    Cryptosporidium is a protozoan parasite of humans and animals and has a worldwide distribution. The parasite has a unique epidemiology in Middle Eastern countries where the IId subtype family of Cryptosporidium parvum dominates. However, there has been no information on Cryptosporidium species in Yemen. Thus, this study was conducted in Yemen to examine the distribution of Cryptosporidium species and subtype families. Fecal samples were collected from 335 patients who attended hospitals in Sana'a city. Cryptosporidium species were determined by PCR and sequence analysis of the 18 s rRNA gene. Cryptosporidium parvum and C. hominis subtypes were identified based on sequence analysis of the 60 kDa glycoprotein (gp60) gene. Out of 335 samples, 33 (9.9%) were positive for Cryptosporidium. Of them, 97% were identified as C. parvum whilst 1 case (3%) was caused by C. hominis. All 7 C. parvum isolates subtyped belonged to the IIaA15G2R1 subtype. The common occurrence of the zoonotic IIa subtype family of C. parvum highlights the potential occurrence of zoonotic transmission of cryptosporidiosis in Yemen. However, this postulation needs confirmation with future molecular epidemiological studies of cryptosporidiosis in both humans and animals in Yemen.

  3. Distribution of Cryptosporidium Genotypes in Storm Event Water Samples from Three Watersheds in New York

    PubMed Central

    Jiang, Jianlin; Alderisio, Kerri A.; Xiao, Lihua

    2005-01-01

    To assess the source and public health significance of Cryptosporidium oocyst contamination in storm runoff, a PCR-restriction fragment length polymorphism technique based on the small-subunit rRNA gene was used in the analysis of 94 storm water samples collected from the Malcolm Brook and N5 stream basins in New York over a 3-year period. The distribution of Cryptosporidium in this study was compared with the data obtained from 27 storm water samples from the Ashokan Brook in a previous study. These three watersheds represented different levels of human activity. Among the total of 121 samples analyzed from the three watersheds, 107 were PCR positive, 101 of which (94.4%) were linked to animal sources. In addition, C. hominis (W14) was detected in six samples collected from the Malcolm Brook over a 2-week period. Altogether, 22 Cryptosporidium species or genotypes were found in storm water samples from these three watersheds, only 11 of which could be attributed to known species/groups of animals. Several Cryptosporidium spp. were commonly found in these three watersheds, including the W1 genotype from an unknown animal source, the W4 genotype from deer, and the W7 genotype from muskrats. Some genotypes were found only in a particular watershed. Aliquots of 113 samples were also analyzed by the Environmental Protection Agency (EPA) Method 1623; 63 samples (55.7%) were positive for Cryptosporidium by microscopy, and 39 (78%) of the 50 microscopy-negative samples were positive by PCR. Results of this study demonstrate that molecular techniques can complement traditional detection methods by providing information on the source of contamination and the human-infective potential of Cryptosporidium oocysts found in water. PMID:16085835

  4. Selection and application of microbial source tracking tools for water-quality investigations

    USGS Publications Warehouse

    Stoeckel, Donald M.

    2005-01-01

    Microbial source tracking (MST) is a complex process that includes many decision-making steps. Once a contamination problem has been defined, the potential user of MST tools must thoroughly consider study objectives before deciding upon a source identifier, a detection method, and an analytical approach to apply to the problem. Regardless of which MST protocol is chosen, underlying assumptions can affect the results and interpretation. It is crucial to incorporate tests of those assumptions in the study quality-control plan to help validate results and facilitate interpretation. Detailed descriptions of MST objectives, protocols, and assumptions are provided in this report to assist in selection and application of MST tools for water-quality investigations. Several case studies illustrate real-world applications of MST protocols over a range of settings, spatial scales, and types of contamination. Technical details of many available source identifiers and detection methods are included as appendixes. By use of this information, researchers should be able to formulate realistic expectations for the information that MST tools can provide and, where possible, successfully execute investigations to characterize sources of fecal contamination to resource waters.

  5. Results From a Microbial Source-Tracking Study at Villa Angela Beach, Cleveland, Ohio, 2007

    USGS Publications Warehouse

    Bushon, Rebecca N.; Stelzer, Erin A.; Stoeckel, Donald M.

    2009-01-01

    During the 2007 recreational season at Villa Angela Beach in Cleveland, Ohio, scientists with the U.S. Geological Survey (USGS) and the Northeast Ohio Regional Sewer District (NEORSD) found high Escherichia coli (E. coli) concentrations that were not easily explained by results obtained to date in ongoing investigations of recreational water quality at the beach. To help understand the sources behind these elevated E. coli concentrations, the USGS and NEORSD sampled beach-area water for Bacteroides DNA markers. Bacteroides are a group of enteric bacteria that are being used in microbial source tracking, in hope that host-associated DNA markers could be used to indicate potential sources of E. coli in the Villa Angela environment. The USGS Ohio Water Microbiology Laboratory analyzed a total of 13 source samples (sewage and waterfowl feces) and 33 beach-area water and sand samples for three Bacteroides DNA markers. This report lists the results of those analyses, along with environmental conditions at Villa Angela on the dates that samples were collected.

  6. F+ RNA coliphage-based microbial source tracking in water resources of South Korea.

    PubMed

    Lee, Jung Eun; Lee, Heetae; Cho, You-Hee; Hur, Hor-Gil; Ko, GwangPyo

    2011-12-15

    We previously demonstrated that genotyping followed by proper statistical analyses of F plus (F+)-specific RNA coliphages can effectively represent fecal origins of either humans or animals. Here, we performed microbial source tracking (MST) using F+ RNA coliphages as a target MST microorganism for identifying fecal sources contaminating ground and surface water in metropolitan Seoul and Gyeonggi Province in South Korea. In total, 71 groundwater and 5 surface water samples were collected and screened for the presence of F+ RNA coliphages. More than 124 F+ coliphages were isolated from six groundwater and five surface water samples by the single agar layer method. F+ RNA coliphages were predominant in both waters (100% and 91%, respectively). Genotyping of 118 F+ RNA coliphages revealed that most (51/60) of the groundwater F+ RNA coliphages belonged to group I, whereas both groups I (25/58) and IV (31/58) were predominantly observed in surface water. Further comparison of phage isolates from human and animal (pig, cow, goose, and chicken) fecal sources using nucleic acid sequencing and principal coordinate analysis showed that groundwater samples formed clusters associated with cow feces, whereas surface waters formed clusters related to chicken and human feces. These results indicate the potential of the F+ RNA coliphage-based MST for identifying fecal contamination sources, which may be further exploited and validated in different geographical regions of the world.

  7. Microbial Source Tracking Markers for Detection of Fecal Contamination in Environmental Waters: Relationships Between Pathogens and Human Health Outcomes

    EPA Science Inventory

    Microbial source tracking (MST) describes a suite of methods and an investigative strategy designed to identify the dominant sources of fecal pollution in environmental waters. The methods rely on the close association of certain fecal microorganisms with a particular host speci...

  8. Cryptosporidium testudinis sp. n., Cryptosporidium ducismarci Traversa, 2010 and Cryptosporidium tortoise genotype III (Apicomplexa: Cryptosporidiidae) in tortoises.

    PubMed

    Jezkova, Jana; Horcickova, Michaela; Hlaskova, Lenka; Sak, Bohumil; Kvetonova, Dana; Novak, Jan; Hofmannova, Lada; McEvoy, John; Kvac, Martin

    2016-10-14

    Understanding of the diversity of species of Cryptosporidium Tyzzer, 1910 in tortoises remains incomplete due to the limited number of studies on these hosts. The aim of the present study was to characterise the genetic diversity and biology of cryptosporidia in tortoises of the family Testudinidae Batsch. Faecal samples were individually collected immediately after defecation and were screened for presence of cryptosporidia by microscopy using aniline-carbol-methyl violet staining, and by PCR amplification and sequence analysis targeting the small subunit rRNA (SSU), Cryptosporidium oocyst wall protein (COWP) and actin genes. Out of 387 faecal samples from 16 tortoise species belonging to 11 genera, 10 and 46 were positive for cryptosporidia by microscopy and PCR, respectively. All samples positive by microscopy were also PCR positive. Sequence analysis of amplified genes revealed the presence of the Cryptosporidium tortoise genotype I (n = 22), C. ducismarci Traversa, 2010 (n = 23) and tortoise genotype III (n = 1). Phylogenetic analyses of SSU, COWP and actin gene sequences revealed that Cryptosporidium tortoise genotype I and C. ducismarci are genetically distinct from previously described species of Cryptosporidium. Oocysts of Cryptosporidium tortoise genotype I, measuring 5.8-6.9 µm × 5.3-6.5 µm, are morphologically distinguishable from C. ducismarci, measuring 4.4-5.4 µm × 4.3-5.3 µm. Oocysts of Cryptosporidium tortoise genotype I and C. ducismarci obtained from naturally infected Russian tortoises (Testudo horsfieldii Gray) were infectious for the same tortoise but not for Reeve's turtles (Mauremys reevesii [Gray]), common garter snake (Thamnophis sirtalis [Linnaeus]), zebra finches (Taeniopygia guttata [Vieillot]) and SCID mice (Mus musculus Linnaeus). The prepatent period was 11 and 6 days post infection (DPI) for Cryptosporidium tortoise genotype I and C. ducismarci, respectively; the patent period was longer than 200 days for both cryptosporidia

  9. Prevalence and Diversity of Cryptosporidium and Giardia Identified Among Feral Pigs in Texas.

    PubMed

    Rodriguez-Rivera, Lorraine D; Cummings, Kevin J; McNeely, Isaac; Suchodolski, Jan S; Scorza, Andrea V; Lappin, Michael R; Mesenbrink, Brian T; Leland, Bruce R; Bodenchuk, Michael J

    2016-12-01

    The population size and geographic range of feral pigs in the United States are rapidly expanding. Nevertheless, the role of this invasive species in the ecology and transmission of zoonotic enteric pathogens is poorly understood. Our objectives were to describe the prevalence and diversity of Cryptosporidium and Giardia shedding among feral pigs throughout Texas and to identify risk factors for infection. Fecal samples were collected from feral pigs in Texas from February 2014 through May 2015. Cryptosporidium oocysts and Giardia cysts were detected using a direct immunofluorescence assay, and genotyping of positive samples was performed. The prevalence of Cryptosporidium shedding was 1.6% (6/370), and C. scrofarum and C. suis were identified. The prevalence of Giardia shedding was 4.3% (16/370), and assemblages A and E were identified. Cryptosporidium shedding was significantly more common among juvenile and subadult pigs than among adult pigs, but age group was not associated with Giardia shedding status. Feral pigs may serve as a source of Cryptosporidium and Giardia transmission to humans and livestock.

  10. Cryptosporidium erinacei and C. parvum in a group of overwintering hedgehogs.

    PubMed

    Hofmannová, Lada; Hauptman, Karel; Huclová, Kristýna; Květoňová, Dana; Sak, Bohumil; Kváč, Martin

    2016-10-01

    This study describes cryptosporidiosis in an overwintering group of 15 European hedgehogs (Erinaceus europaeus), comprising 3 adults and 12 juveniles. Four juvenile hedgehogs were hospitalised with anorexia, malodorous diarrhoea and dehydration. Immediate parasitological examinations revealed the presence of Cryptosporidium sp. in these animals and also in 5 other juveniles. All hedgehogs were coproscopically monitored for 4 months over the winter season. Shedding of Cryptosporidium oocysts persisted from 6 to 70 days. Repeated shedding of Cryptosporidium oocysts occurred in 3 animals after 4 months subsequent to the first outbreak. Clinical signs were observed only at the beginning of the outbreak (apathy, anorexia, general weakness, mild dehydration, and malodorous faeces with changed consistence - soft/diarrhoea) in the 4 hospitalised juveniles. Overall 11 hedgehogs were Cryptosporidium-positive, both microscopically and by PCR methods. Sequence analyses of SSU rRNA and gp60 genes revealed the presence of C. parvum IIdA18G1 subtype in all positive hedgehogs. Moreover, 3 hedgehogs had a mixed infection of the zoonotic C. parvum and C. erinacei XIIIaA19R13 subtype. Cryptosporidium infections can be rapidly spread among debilitated animals and the positive hedgehogs released back into the wild can be a source of the infection for individuals weakened after hibernation.

  11. Molecular Investigations of Bacteroides as Microbial Source Tracking Tools in Southeast Louisiana Watersheds

    NASA Astrophysics Data System (ADS)

    Schulz, C. J.; Childers, G. W.; Engel, A. S.

    2006-12-01

    Microbial Source Tracking (MST) is a developing field that is gaining increased attention. MST refers to a host of techniques that discriminates among the origins of fecal material found in natural waters from different sources (e.g. human, livestock, and wildlife) by using microbial indicator species with specificity to only certain host organisms. The development of species-specific molecular markers would allow for better evaluation of public health risks and tracking of nutrient sources impacting a watershed. Although several MST methods have been reported with varying levels of success, few offer general applicability for natural waters due to spatial and temporal constraints associated with these methods. One group of molecular MST markers that show promise for broad environmental applications are molecular 16S rDNA probes for Bacteroides. This method is based on 16S rDNA detection directly from environmental samples without the need for a preliminary cultivation step. In this study we have expanded previous sampling efforts to compile a database of over 1000 partial 16S rRNA Bacteroides genes retrieved from the fecal material of 15 different host species (human, cat, dog, pig, kangaroo). To characterize survival of Bacteroides outside of the host, survival time of the Bacteroides marker was compared to that of E.coli under varying natural environmental conditions (temperature and salinity). Bacteroides displayed a survival curve with shouldering and tailing similar to that of E.coli, but log reduction times differed with treatment. In summary, MST marker stability was identified within host species and the overall Bacteroides community structure correlated to host diet, suggesting that detection of a Bacteroides community could confidently identify fecal contamination point sources. Natural water samples from southeast Louisiana were collected for MST including the Tangipahoa River watershed where the source of fecal contamination has been hotly debated. The

  12. Classification Tree Method for Bacterial Source Tracking with Antibiotic Resistance Analysis Data

    PubMed Central

    Price, Bertram; Venso, Elichia A.; Frana, Mark F.; Greenberg, Joshua; Ware, Adam; Currey, Lee

    2006-01-01

    Various statistical classification methods, including discriminant analysis, logistic regression, and cluster analysis, have been used with antibiotic resistance analysis (ARA) data to construct models for bacterial source tracking (BST). We applied the statistical method known as classification trees to build a model for BST for the Anacostia Watershed in Maryland. Classification trees have more flexibility than other statistical classification approaches based on standard statistical methods to accommodate complex interactions among ARA variables. This article describes the use of classification trees for BST and includes discussion of its principal parameters and features. Anacostia Watershed ARA data are used to illustrate the application of classification trees, and we report the BST results for the watershed. PMID:16672492

  13. Tracking sources of bacterial contamination in stormwater discharges to Mission Bay, California.

    PubMed

    Schiff, K; Kinney, P

    2001-01-01

    Sources of the indicator bacteria total coliform, fecal coliform, and enterococcus were investigated in stormwater flows discharging to Mission Bay, a heavily used aquatic park in San Diego, California. Stormwater flows were targeted because long-term receiving water monitoring of the bay indicated that wet weather discharges were the predominant source of bacterial contamination. Exceedences in water quality objectives for body contact recreation established by the State of California most often occurred in the east bay, where the least amount of circulation and largest quantities of stormwater discharges occur. Unlike the wet weather results, almost all of the 89 storm drains that discharge to the bay either did not have flowing fresh water or did not contain exceedingly high bacteria densities during dry weather. Upstream tracking during multiple storm events on two of the largest watersheds draining to the bay showed that sources of indicator bacteria were diffuse and widespread. Densities were as high at the head of each watershed as they were at the mouth, where both discharged to the bay. Every reach in each creek exceeded State of California water quality objectives and had densities similar to surface flows measured before they entered the separate municipal storm sewer system from urban land uses, such as residential, commercial, and industrial, as well as open lands.

  14. Particle filtering for arrival time tracking in space and source localization.

    PubMed

    Michalopoulou, Zoi-Heleni; Jain, Rashi

    2012-11-01

    Locating and tracking a source in an ocean environment and estimating environmental parameters of a sound propagation medium are critical tasks in ocean acoustics. Many approaches for both are based on full field calculations which are computationally intensive and sensitive to assumptions on the structure of the environment. Alternative methods that use only select features of the acoustic field for localization and environmental parameter estimation have been proposed. The focus of this paper is the development of a method that extracts arrival times and amplitudes of distinct paths from measured acoustic time-series using sequential Bayesian filtering, namely, particle filtering. These quantities, along with complete posterior probability density functions, also extracted by filtering, are employed in source localization and bathymetry estimation. Aspects of the filtering methodology are presented and studied in terms of their impact on the uncertainty in the arrival time estimates. Using the posterior probability densities of arrival times, source localization and water depth estimation are performed for the Haro Strait Primer experiment; the results are compared to those of conventional methods. The comparison demonstrates a significant advantage in the proposed approach.

  15. Analysis of human and animal fecal microbiota for microbial source tracking.

    PubMed

    Lee, Jung Eun; Lee, Sunghee; Sung, Joohon; Ko, GwangPyo

    2011-02-01

    Microbial compositions of human and animal feces from South Korea were analyzed and characterized. In total, 38 fecal samples (14 healthy adult humans, 6 chickens, 6 cows, 6 pigs and 6 geese) were analyzed by 454 pyrosequencing of the V2 region of the 16S rRNA gene. Four major phyla, Actinobacteria, Proteobacteria, Firmicutes and Bacteroidetes, were identified in the samples. Principal coordinate analysis suggested that microbiota from the same host species generally clustered, with the exception of those from humans, which exhibited sample-specific compositions. A network-based analysis revealed that several operational taxonomic units (OTUs), such as Lactobacillus sp., Clostridium sp. and Prevotella sp., were commonly identified in all fecal sources. Other OTUs were present only in fecal samples from a single organism. For example, Yania sp. and Bifidobacterium sp. were identified specifically in chicken and human fecal samples, respectively. These specific OTUs or their respective biological markers could be useful for identifying the sources of fecal contamination in water by microbial source tracking.

  16. Identifying the origin of waterbird carcasses in Lake Michigan using a neural network source tracking model

    USGS Publications Warehouse

    Kenow, Kevin P.; Ge, Zhongfu; Fara, Luke J.; Houdek, Steven C.; Lubinski, Brian R.

    2016-01-01

    Avian botulism type E is responsible for extensive waterbird mortality on the Great Lakes, yet the actual site of toxin exposure remains unclear. Beached carcasses are often used to describe the spatial aspects of botulism mortality outbreaks, but lack specificity of offshore toxin source locations. We detail methodology for developing a neural network model used for predicting waterbird carcass motions in response to wind, wave, and current forcing, in lieu of a complex analytical relationship. This empirically trained model uses current velocity, wind velocity, significant wave height, and wave peak period in Lake Michigan simulated by the Great Lakes Coastal Forecasting System. A detailed procedure is further developed to use the model for back-tracing waterbird carcasses found on beaches in various parts of Lake Michigan, which was validated using drift data for radiomarked common loon (Gavia immer) carcasses deployed at a variety of locations in northern Lake Michigan during September and October of 2013. The back-tracing model was further used on 22 non-radiomarked common loon carcasses found along the shoreline of northern Lake Michigan in October and November of 2012. The model-estimated origins of those cases pointed to some common source locations offshore that coincide with concentrations of common loons observed during aerial surveys. The neural network source tracking model provides a promising approach for identifying locations of botulinum neurotoxin type E intoxication and, in turn, contributes to developing an understanding of the dynamics of toxin production and possible trophic transfer pathways.

  17. DETECTION OF CRYPTOSPORIDIUM OOCYSTS IN WATER MATRICES

    EPA Science Inventory

    Since the advent and recognition of waterborne outbreaks of cryptosporidiosis great effort has been expended on development of methods for detecting Cryptosporidium oocysts in water. Oocysts recovery rates using a method originally developed for detecting Giardia cysts ranged fr...

  18. Cryptosporidium in birds, fish and amphibians.

    PubMed

    Ryan, Una

    2010-01-01

    Whilst considerable information is available for avian cryptosporidiosis, scant information is available for Cryptosporidium infections in fish and amphibians. The present review details recent studies in avian cryptosporidiosis and our current knowledge of piscine and amphibian infections.

  19. INACTIVATION OF CRYPTOSPORIDIUM PARVUM OOCYSTS WITH OZONE

    EPA Science Inventory

    Ozone inactivation rates for Cryptosporidium parvum (C. parvum) oocysts were determined with an in-vitro excystation method based on excysted sporozoite counts. Results were consistent with published animal infectivity data for the same C. parvum strain. The inactivation kinetics...

  20. AN EVALUATION OF CRYPTOSPORIDIUM PARVUM GENOTYPING

    EPA Science Inventory

    We evaluated the specificity and sensitivity of 11 previously described species differentiation and genotyping PCR protocols for detection of Cryptosporidium parasites. Genomic DNA from three species of Crytosporidium parasites (genotype 1 and genotype 2 of C. parvum, C. muris, a...

  1. Source tracking of prokaryotic communities in fermented grain of Chinese strong-flavor liquor.

    PubMed

    Wang, Xueshan; Du, Hai; Xu, Yan

    2017-03-06

    The fermentation process of Chinese strong-flavor liquor involves numerous microbes originating from Daqu and pit mud. Daqu is the starter of fermentation, and pit mud acts as another source of inoculum of microbes in the liquor-making process. However, the contribution of microbes in pit mud and Daqu to fermented grain, and the sources of microbes in fermented grain are still waiting to be defined clearly. In this study, prokaryotic communities in fermented grain, pit mud and Daqu were identified via next generation sequencing of the V4 region of 16S rRNA gene. Principal-coordinate analysis indicated that Daqu had stronger influence on the prokaryotic communities in fermented grain at the prophase of fermentation, but pit mud influenced the fermented grain continuously during the whole fermentation process. Totally, 299 genera were detected in all fermented grain, pit mud and Daqu samples. Among them, 204 genera were detected in 3days' fermented grain. Ten genera (Lactobacillus, Leuconostoc, Staphylococcus, Gluconobacter, Acetobacter, Petrimonas, Clostridium, Ruminococcus, Methanobacterium and Methanobrevibacter) were dominant, and accounted for 84.31%-87.13% relative abundance of the total prokaryotic community in fermented grain. Venn analysis indicated Daqu was the main source of strict aerobes and facultative aerobes, which took up over 74% of prokaryotic communities in fermented grain. Conversely, pit mud was the sustained-release source of anaerobes, which accounted for over 14% of prokaryotic communities in fermented grain. In addition, part of anaerobes originated from both Daqu and pit mud. This study could help track the source of prokaryotic communities in fermented grain, and improve the quality and controllability in liquor production.

  2. Use of Bacteroidales microbial source tracking to monitor fecal contamination in fresh produce production.

    PubMed

    Ravaliya, Kruti; Gentry-Shields, Jennifer; Garcia, Santos; Heredia, Norma; Fabiszewski de Aceituno, Anna; Bartz, Faith E; Leon, Juan S; Jaykus, Lee-Ann

    2014-01-01

    In recent decades, fresh and minimally processed produce items have been associated with an increasing proportion of food-borne illnesses. Most pathogens associated with fresh produce are enteric (fecal) in origin, and contamination can occur anywhere along the farm-to-fork chain. Microbial source tracking (MST) is a tool developed in the environmental microbiology field to identify and quantify the dominant source(s) of fecal contamination. This study investigated the utility of an MST method based on Bacteroidales 16S rRNA gene sequences as a means of identifying potential fecal contamination, and its source, in the fresh produce production environment. The method was applied to rinses of fresh produce, source and irrigation waters, and harvester hand rinses collected over the course of 1 year from nine farms (growing tomatoes, jalapeño peppers, and cantaloupe) in Northern Mexico. Of 174 samples, 39% were positive for a universal Bacteroidales marker (AllBac), including 66% of samples from cantaloupe farms (3.6 log10 genome equivalence copies [GEC]/100 ml), 31% of samples from tomato farms (1.7 log10 GEC/100 ml), and 18% of samples from jalapeño farms (1.5 log10 GEC/100 ml). Of 68 AllBac-positive samples, 46% were positive for one of three human-specific markers, and none were positive for a bovine-specific marker. There was no statistically significant correlation between Bacteroidales and generic Escherichia coli across all samples. This study provides evidence that Bacteroidales markers may serve as alternative indicators for fecal contamination in fresh produce production, allowing for determination of both general contamination and that derived from the human host.

  3. Microbial Source Tracking as a Tool for TMDL Development, Little Blue River in Independence, Missouri

    USGS Publications Warehouse

    Christensen, Eric D.; Bushon, Rebecca N.; Brady, Amie M.G.

    2013-01-01

    The Little Blue River in Jackson County, Missouri has been listed by the Missouri Department of Natural Resources as impaired by bacteria for the protection of aquatic life and contact recreation from urban point and nonpoint sources. The Clean Water Act requires that a total maximum daily load (TMDL) for Escherichia coli (E. coli) be developed. Over a 5-year period, 108 base-flow, 87 stormflow, 48 fecal source, and 12 sewage influent samples were collected and analyzed for E. coli and Bacteroides general and host-associated microbial source tracking (MST) genetic markers. Less than half of the main-stem base-flow samples exceeded the E. coli state standard, whereas, all of the stormflow samples exceeded the standard during the recreation season (April through October). Both E. coli and MST markers were detected more frequently and at higher concentrations in stormflow samples. Only 14 percent of samples with E. coli detections greater than the Missouri state standard of 206 colonies per 100 milliliters had the human-associated Bacteroides marker as the only identified marker; therefore, Little Blue River TMDL development may require a broader scope beyond the municipal separate storm sewer system if bacteria sources are to be identified and addressed. Fecal samples showed a greater specificity with the human-associated marker than the dog- or ruminant-associated Bacteroides markers; however, false positives were at least one order of magnitude lower than true positives. MST data may be a useful tool for identifying probable sources of contamination and directing TMDL strategies.

  4. Targeted sampling protocol as prelude to bacterial source tracking with Enterococcus faecalis.

    PubMed

    Kuntz, Robin L; Hartel, Peter G; Godfrey, Dominique G; McDonald, Jennifer L; Gates, Keith W; Segars, William I

    2003-01-01

    Recent studies suggest that host origin databases for bacterial source tracking (BST) must contain a large number of isolates because bacterial subspecies change with geography and time. A new targeted sampling protocol was developed as a prelude to BST to minimize these changes. The research was conducted on the Sapelo River, a tidal river on the Georgia coast. A general sampling of the river showed fecal enterococcal numbers ranging from <10 (below the limit of detection) to 990 colony-forming units (CFU) per 100 mL. Locations with high enterococcal numbers were combined with local knowledge to determine targeted sampling sites. Fecal enterococcal numbers around one site ranged from <10 to 24,000 CFU per 100 mL. Bacterial source tracking was conducted to determine if a wastewater treatment facility at the site was responsible for this contamination. The fecal indicator bacterium was Enterococcus faecalis. Ribotyping, automated with a RiboPrinter (DuPont Qualicon, Wilmington, DE), was the BST method. Thirty-seven ribotypes were observed among 83 Ent. faecalis isolates obtained from the Sapelo River and the wastewater lagoon. Sixteen ribotypes were associated with either the river or the lagoon, and only five ribotypes (14%) were shared. Nevertheless, these five ribotypes represented 39 of the 83 Ent. faecalis isolates, almost a majority (47%). These results suggest that the fecal contamination in the river came from the wastewater treatment facility. As a prelude to BST, targeted sampling minimized subspecies changes with geography and time, and eliminated the need for a permanent host origin database by restricting BST to a small geographic area and requiring sampling to be completed in one day.

  5. Prevalence of Microsporidia, Cryptosporidium spp., and Giardia spp. in beavers (Castor canadensis) in Massachusetts

    USGS Publications Warehouse

    Fayer, R.; Santin, M.; Trout, J.M.; DeStefano, S.; Koenen, K.; Kaur, T.

    2006-01-01

    Feces from 62 beavers (Castor canadensis) in Massachusetts were examined by fluorescence microscopy (IFA) and polymerase chain reaction (PCR) for Microsporidia species, Cryptosporidium spp., and Giardia spp. between January 2002 and December 2004. PCR-positive specimens were further examined by gene sequencing. Protist parasites were detected in 6.4% of the beavers. All were subadults and kits. Microsporidia species were not detected. Giardia spp. was detected by IFA from four beavers; Cryptosporidium spp. was also detected by IFA from two of these beavers. However, gene sequence data for the ssrRNA gene from these two Cryptosporidium spp.-positive beavers were inconclusive in identifying the species. Nucleotide sequences of the TPI, ssrRNA, and ??-giardin genes for Giardia spp. (deposited in GenBank) indicated that the four beavers were excreting Giardia duodenalis Assemblage B, the zoonotic genotype representing a potential source of waterborne Giardia spp. cysts. Copyright 2006 by American Association of Zoo Veterinarians.

  6. Prevalence of Microsporidia, Cryptosporidium spp., and Giardia spp. in beavers (Castor canadensis) in Massachusetts.

    PubMed

    Fayer, Ronald; Santín, Mónica; Trout, James M; DeStefano, Stephen; Koenen, Kiana; Kaur, Taranjit

    2006-12-01

    Feces from 62 beavers (Castor canadensis) in Massachusetts were examined by fluorescence microscopy (IFA) and polymerase chain reaction (PCR) for Microsporidia species, Cryptosporidium spp., and Giardia spp. between January 2002 and December 2004. PCR-positive specimens were further examined by gene sequencing. Protist parasites were detected in 6.4% of the beavers. All were subadults and kits. Microsporidia species were not detected. Giardia spp. was detected by IFA from four beavers; Cryptosporidium spp. was also detected by IFA from two of these beavers. However, gene sequence data for the ssrRNA gene from these two Cryptosporidium spp.-positive beavers were inconclusive in identifying the species. Nucleotide sequences of the TPI, ssrRNA, and beta-giardin genes for Giardia spp. (deposited in GenBank) indicated that the four beavers were excreting Giardia duodenalis Assemblage B, the zoonotic genotype representing a potential source of waterborne Giardia spp. cysts.

  7. Spinacia oleracea L. leaf stomata harboring Cryptosporidium parvum oocysts: a potential threat to food safety.

    PubMed

    Macarisin, Dumitru; Bauchan, Gary; Fayer, Ronald

    2010-01-01

    Cryptosporidium parvum is a cosmopolitan microscopic protozoan parasite that causes severe diarrheal disease (cryptosporidiosis) in mammals, including humans and livestock. There is growing evidence of Cryptosporidium persistence in fresh produce that may result in food-borne infection, including sporadic cases as well as outbreaks. However, drinking and recreational waters are still considered the major sources of Cryptosporidium infection in humans, which has resulted in prioritization of studies of parasite etiology in aquatic environments, while the mechanisms of transmission and parasite persistence on edible plants remain poorly understood. Using laser scanning confocal microscopy together with fluorescein-labeled monoclonal antibodies, C. parvum oocysts were found to strongly adhere to spinach plants after contact with contaminated water, to infiltrate through the stomatal openings in spinach leaves, and to persist at the mesophyll level. These findings and the fact that this pathogenic parasite resists washing and disinfection raise concerns regarding food safety.

  8. Quantitative health risk assessment of Cryptosporidium in rivers of southern China based on continuous monitoring.

    PubMed

    An, Wei; Zhang, Dongqing; Xiao, Shumin; Yu, Jianwei; Yang, Min

    2011-06-01

    The concentrations of Cryptosporidium in the source water of several cities of Zhejiang Province, China were determined to be in the range of 0-17 oocysts/10 L in the rainy season in 2008, with a mean value of 7 oocysts/10 L. Based on the investigation data, comprehensive risk assessment of Cryptosporidium infection was performed by considering different water intake routes as well as water consumption. Intakes of unboiled tapwater (including drinking and tooth-brushing and food and dish washing) and source water (through swimming in rivers) were estimated to be 2.59-25.9 and 0.32-0.74 L/year-person, respectively. The mortality due to Cryptosporidium infection for people in this region, excluding HIV-infected patients, was calculated as 0-0.0146 per 10(5) persons using a conditional probability formula. Disability-adjusted life years (DALYs) were used to quantify the risk of Cryptosporidium infection, for which uncertainty was analyzed. For people who consumed conventionally treated water, the DALYs due to Cryptosporidium infection were 6.51 per 10(5) (95% CI: 2.16 × 10(-5)-22.35 × 10(-5)) persons, which were higher than a risk judged acceptable by some (1.97 × 10(-5) DALYs per year), and the risk for those consuming ozone-treated water became 0.0689 × 10(-5) DALYs per year. The major risk of infection resulted from swimming in the river. This study provides a method to establish the risk of Cryptosporidium infection and optimize the scheme for reducing the risk effectively, which is useful for the modification of water quality standards based on cost utility analysis given use of DALYs.

  9. Cryptosporidium Propidium Monoazide-PCR, a Molecular Biology-Based Technique for Genotyping Viable Cryptosporidium Oocysts

    EPA Science Inventory

    Cryptosporidium is an important waterborne protozoan parasite that can cause severe diarrhea and death in the immunocompromised. Current methods to monitor for Cryptosporidium oocysts in water are microscopy-based USEPA Methods 1622 and 1623. These methods assess total levels o...

  10. Inorganic nitrogen, sterols and bacterial source tracking as tools to characterize water quality and possible contamination sources in surface water.

    PubMed

    Furtula, Vesna; Osachoff, Heather; Derksen, George; Juahir, Hafizan; Colodey, Al; Chambers, Patricia

    2012-03-15

    The effects of agricultural activities on stream water quality were assessed by nitrogen analysis, further investigated by gas chromatography mass spectrometry (GC-MS) sterol analysis (including chemometric analysis), and characterized by bacterial source tracking (BST). Surface water samples were collected from five sites, throughout the agriculturally-influenced Nathan Creek watershed, British Columbia, Canada and a nearby control site between October 2005 and March 2006. From a total of 48 samples, Canadian Water Quality Guidelines were exceeded nineteen times for nitrate (NO3-; guideline value: 2.94 mg/L N) and four times for un-ionized ammonia (NH3; guideline value 0.019 mg/L N). Gas chromatography mass spectrometry single ion monitoring (GC-MS SIM) analysis of 18 sterols showed that five fecal sterols (coprostanol, episoprostanol, cholesterol, cholestanol, desmosterol) were detected at all sites except the control site (where only cholesterol, cholestanol and desmosterol were detected). Three phytosterols (campesterol, stigmasterol and β-sitosterol) were also detected at all sites while the hormone estrone was present at one site on two occasions at concentrations of 0.01 and 0.04 μg/L. Chemometric analysis (principal component analysis and cluster analysis) grouped sites based on their similarities in sterol composition. Analysis of ten sterol ratios (seven for identifying human fecal contamination and four for differentiating sources of fecal contamination) showed multiple instances of human and animal contamination for every site but the control site. Application of a Bacteroides-BST method confirmed contamination from ruminant animals, pigs and dogs in varying combinations at all impact sites. Together, these results confirmed the impact of agricultural activities on the Nathan Creek watershed and support a need for better land management practices to protect water quality and aquatic life.

  11. Response of TLD-albedo and nuclear track dosimeters exposed to plutonium sources

    SciTech Connect

    Brackenbush, L.W.; Baumgartner, W.V.; Fix, J.J.

    1991-12-01

    Neutron dosimetry has been extensively studied at Hanford since the mid-1940s. At the present time, Hanford contractors use thermoluminescent dosimeter (TLD)-albedo dosimeters to record the neutron dose equivalent received by workers. The energy dependence of the TLD-albedo dosimeter has been recognized and documented since introduced at Hanford in 1964 and numerous studies have helped assure the accuracy of dosimeters. With the recent change in Hanford`s mission, there has been a significant decrease in the handling of plutonium tetrafluoride, and an increase in the handling of plutonium metal and plutonium oxide sources. This study was initiated to document the performance of the current Hanford TLD-albedo dosimeter under the low scatter conditions of the calibration laboratory and under the high scatter conditions in the work place under carefully controlled conditions at the Plutonium Finishing Plant (PFP). The neutron fields at the PFP facility were measured using a variety of instruments, including a multisphere spectrometer, tissue equivalent proportional counters, and specially calibrated rem meters. Various algorithms were used to evaluate the TLD-albedo dosimeters, and the results are given in this report. Using current algorithms, the dose equivalents evaluated for bare sources and sources with less than 2.5 cm (1 in.) of acrylic plastic shielding in high scatter conditions typical of glove box operations are reasonably accurate. Recently developed CR-39 track etch dosimeters (TEDs) were also exposed in the calibration laboratory and at the PFP. The results indicate that the TED dosimeters are quite accurate for both bare and moderated neutron sources. Until personnel dosimeter is available that incorporates a direct measure of the neutron dose to a person, technical uncertainties in the accuracy of the recorded data will continue.

  12. Response of TLD-albedo and nuclear track dosimeters exposed to plutonium sources

    SciTech Connect

    Brackenbush, L.W.; Baumgartner, W.V.; Fix, J.J.

    1991-12-01

    Neutron dosimetry has been extensively studied at Hanford since the mid-1940s. At the present time, Hanford contractors use thermoluminescent dosimeter (TLD)-albedo dosimeters to record the neutron dose equivalent received by workers. The energy dependence of the TLD-albedo dosimeter has been recognized and documented since introduced at Hanford in 1964 and numerous studies have helped assure the accuracy of dosimeters. With the recent change in Hanford's mission, there has been a significant decrease in the handling of plutonium tetrafluoride, and an increase in the handling of plutonium metal and plutonium oxide sources. This study was initiated to document the performance of the current Hanford TLD-albedo dosimeter under the low scatter conditions of the calibration laboratory and under the high scatter conditions in the work place under carefully controlled conditions at the Plutonium Finishing Plant (PFP). The neutron fields at the PFP facility were measured using a variety of instruments, including a multisphere spectrometer, tissue equivalent proportional counters, and specially calibrated rem meters. Various algorithms were used to evaluate the TLD-albedo dosimeters, and the results are given in this report. Using current algorithms, the dose equivalents evaluated for bare sources and sources with less than 2.5 cm (1 in.) of acrylic plastic shielding in high scatter conditions typical of glove box operations are reasonably accurate. Recently developed CR-39 track etch dosimeters (TEDs) were also exposed in the calibration laboratory and at the PFP. The results indicate that the TED dosimeters are quite accurate for both bare and moderated neutron sources. Until personnel dosimeter is available that incorporates a direct measure of the neutron dose to a person, technical uncertainties in the accuracy of the recorded data will continue.

  13. Cryptosporidium ubiquitum n.sp. in animals and humans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A new species, Cryptosporidium ubiquitum, previously identified as the Cryptosporidium cervine genotype is described. In published studies the cervine genotype was reported in wild and domesticated ruminants, rodents, carnivores, and primates including humans. Molecular data for C. ubiquitum have b...

  14. Examining Signal Decomposition in Ge Tracking Detectors through Source-Based Coincidence Measurements

    NASA Astrophysics Data System (ADS)

    Cromaz, M.; Campbell, C. M.; Clark, R. M.; Crawford, H. L.; Fallon, P.; Lee, I. Y.; Macchiavelli, A. O.; Wiens, A.; Riley, L.; Taniuchi, R.

    2016-03-01

    The performance of a gamma-ray tracking detector, such as those used in the GRETINA spectrometer, is dependent on its ability to accurately locate multiple interaction points in the Ge crystal. Interactions are located by observing both net and induced charge as a function of time on the detector's segmented contact. As multiple interactions are likely, linear combinations of basis signals, a set of simulated signals with unit charge deposited on a grid that spans the detector volume, are fit against the observed signal yielding the interaction positions. While the location of the primary interaction point was found to be good (σpos <= 2 mm) the location of secondary, lower energy interactions appear less reliable. To investigate this issue, we carried out a series of source-based coincidence measurements. These employed a collimated source and a secondary detector by which we could select single interaction events. Given these events originate from known positions, we can take them in combination to directly test the efficacy of the signal decomposition procedure. We will present a description of the method and preliminary results with a GRETINA quad detector. This work is supported by the U.S. Department of Energy under Contract No. DE-AC02-05CHI1231.

  15. Performance Evaluation of Block Acquisition and Tracking Algorithms Using an Open Source GPS Receiver Platform

    NASA Technical Reports Server (NTRS)

    Ramachandran, Ganesh K.; Akopian, David; Heckler, Gregory W.; Winternitz, Luke B.

    2011-01-01

    Location technologies have many applications in wireless communications, military and space missions, etc. US Global Positioning System (GPS) and other existing and emerging Global Navigation Satellite Systems (GNSS) are expected to provide accurate location information to enable such applications. While GNSS systems perform very well in strong signal conditions, their operation in many urban, indoor, and space applications is not robust or even impossible due to weak signals and strong distortions. The search for less costly, faster and more sensitive receivers is still in progress. As the research community addresses more and more complicated phenomena there exists a demand on flexible multimode reference receivers, associated SDKs, and development platforms which may accelerate and facilitate the research. One of such concepts is the software GPS/GNSS receiver (GPS SDR) which permits a facilitated access to algorithmic libraries and a possibility to integrate more advanced algorithms without hardware and essential software updates. The GNU-SDR and GPS-SDR open source receiver platforms are such popular examples. This paper evaluates the performance of recently proposed block-corelator techniques for acquisition and tracking of GPS signals using open source GPS-SDR platform.

  16. Pinpointing Production: A Particle Tracking Simulator Identifies Source Regions of Escaping Planetary O+

    NASA Astrophysics Data System (ADS)

    Johnson, B. C.

    2015-12-01

    As observations of current Martian atmospheric erosion rates become increasingly precise, a complete understanding of the time evolution of the Martian atmosphere will be limited not by incomplete knowledge of current loss rates, but by an incomplete understanding of how this loss rate changes over time. Making progress on this front necessarily requires detailed understanding of the various acceleration mechanisms that lead to escape. With this in mind, we note that one open question which cannot be answered by in situ measurements of escaping ions is the altitude from which these escaping ions originate. A 3-D kinetic model, the Adaptive Mesh Particle Simulator (AMPS), is used to track oxygen ions from their source. AMPS takes collisions into account, allowing for the examination of the entire upper atmosphere, from beneath the ionospheric peak, through the transition region in which collisions become rare, all the way out to several Mars radii, making AMPS an effective tool for studying the source regions associated with the escape of O+ from the atmosphere of Mars.

  17. The EnzymeTracker: an open-source laboratory information management system for sample tracking

    PubMed Central

    2012-01-01

    Background In many laboratories, researchers store experimental data on their own workstation using spreadsheets. However, this approach poses a number of problems, ranging from sharing issues to inefficient data-mining. Standard spreadsheets are also error-prone, as data do not undergo any validation process. To overcome spreadsheets inherent limitations, a number of proprietary systems have been developed, which laboratories need to pay expensive license fees for. Those costs are usually prohibitive for most laboratories and prevent scientists from benefiting from more sophisticated data management systems. Results In this paper, we propose the EnzymeTracker, a web-based laboratory information management system for sample tracking, as an open-source and flexible alternative that aims at facilitating entry, mining and sharing of experimental biological data. The EnzymeTracker features online spreadsheets and tools for monitoring numerous experiments conducted by several collaborators to identify and characterize samples. It also provides libraries of shared data such as protocols, and administration tools for data access control using OpenID and user/team management. Our system relies on a database management system for efficient data indexing and management and a user-friendly AJAX interface that can be accessed over the Internet. The EnzymeTracker facilitates data entry by dynamically suggesting entries and providing smart data-mining tools to effectively retrieve data. Our system features a number of tools to visualize and annotate experimental data, and export highly customizable reports. It also supports QR matrix barcoding to facilitate sample tracking. Conclusions The EnzymeTracker was designed to be easy to use and offers many benefits over spreadsheets, thus presenting the characteristics required to facilitate acceptance by the scientific community. It has been successfully used for 20 months on a daily basis by over 50 scientists. The EnzymeTracker is

  18. Mussels (Perna perna) as bioindicator of environmental contamination by Cryptosporidium species with zoonotic potential

    PubMed Central

    Mariné Oliveira, Geisi Ferreira; do Couto, Melissa Carvalho Machado; de Freitas Lima, Marcelo; do Bomfim, Teresa Cristina Bergamo

    2016-01-01

    Sources of contamination such as animal feces runoff, organic fertilizer application, and the release of partially treated or untreated sewage can lead to the contamination of aquatic environments by Cryptosporidium spp. The quality of mussels as food is closely related to the sanitary conditions of the marine environment where these bivalves are found. Marine mollusks are filter feeders that are able to retain Cryptosporidium oocysts in their tissue, thus functioning as bioindicators. A total of 72 pooled mussel samples of the species Perna perna were collected at two sites (A and B) in the municipality of Mangaratiba, Rio de Janeiro State, Brazil. Sampling involved removal of 30 mussels, from each collection site every month for one year. The 30 mussels from each sampling were then allocated into three groups of 10. Two Cryptosporidium spp. genes (18S and GP60) were targeted for DNA amplification from the samples obtained. After purification, all of the products obtained were sequenced and phylogenetic analyses were performed. Of the 72 samples analyzed using the nested-PCR for the 18S gene target, 29.2% were positive for the presence of Cryptosporidium spp. Of these samples, 52.4% were collected at site A (ie 11/21) and 47.6% at site B (ie 10/21). The 18S genes of all the samples considered positive for Cryptosporidium spp. were sequenced, and the following three species were identified: Cryptosporidium parvum, C. meleagridis, and C. andersoni. Three distinct C. parvum subtypes (IIaA19G2R2; IIaA20G2R2; IIaA20G3R2) were identified using the GP60 gene. More studies to evaluate the zoonotic potential of this species should be performed as both sampling locations contain human and/or animal fecal contaminants. PMID:26977402

  19. Microbial source tracking in shellfish harvesting waters in the Gulf of Nicoya, Costa Rica.

    PubMed

    Symonds, E M; Young, S; Verbyla, M E; McQuaig-Ulrich, S M; Ross, E; Jiménez, J A; Harwood, V J; Breitbart, M

    2017-03-15

    Current microbial water quality monitoring is generally limited to culture-based measurements of fecal indicator bacteria (FIB). Given the many possible sources of fecal pollution within a watershed and extra-intestinal FIB reservoirs, it is important to determine source(s) of fecal pollution as a means to improve water quality and protect public health. The principal objective of this investigation was to characterize the microbial water quality of shellfish harvesting areas in the Gulf of Nicoya, Costa Rica during 2015. In order to achieve this objective, the specificity and sensitivity of 11 existing microbial source tracking (MST) PCR assays, associated with cows (BacCow), dogs (BacCan, DogBac), domestic wastewater (PMMoV), general avian (GFD), gulls (Gull2), horses (HorseBac, HoF), humans (HF183, HPyV), and pigs (PF), were evaluated using domestic wastewater and animal fecal samples collected from the region. The sensitivity of animal-associated assays ranged from 13 to 100%, while assay specificity ranged from 38 to 100%. The specificity of pepper mild mottle virus (PMMoV) and human polyomavirus (HPyV) was 100% for domestic wastewater, as compared to 94% specificity of the HF183 Bacteroidales marker. PMMoV was identified as a useful domestic wastewater-associated marker, with concentrations as high as 1.1 × 10(5) copies/ml and 100% sensitivity and specificity. Monthly surface water samples collected from four shellfish harvesting areas were analyzed using culture-based methods for Escherichia coli as well as molecular methods for FIB and a suite of MST markers, which were selected for their specificity in the region. While culturable E. coli results suggested possible fecal pollution during the monitoring period, the absence of human/domestic wastewater-associated markers and low FIB concentrations determined using molecular methods indicated sufficient microbial water quality for shellfish harvesting. This is the first study to our knowledge to test the

  20. Source Tracking of Nitrous Oxide using A Quantum Cascade Laser System in the Field and Laboratory Environments

    EPA Science Inventory

    Nitrous oxide is an important greenhouse gas and ozone depleting substance. Nitrification and denitrification are two major biological pathways that are responsible for soil emissions of N2O. However, source tracking of in-situ or laboratory N2O production is still challenging to...

  1. Evaluation of the repeatability and reproducibility of a suite of qPCR based microbial source tracking methods

    EPA Science Inventory

    Many PCR-based methods for microbial source tracking (MST) have been developed and validated within individual research laboratories. Inter-laboratory validation of these methods, however, has been minimal, and the effects of protocol standardization regimes have not been thor...

  2. Decay of Fecal Indicator Bacterial Populations and Bovine-Associated Source-Tracking Markers in Freshly Deposited Cow Pats

    EPA Science Inventory

    Understanding the survival of fecal indicator bacteria (FIB) and microbial source-tracking (MST) markers is critical to developing pathogen fate and transport models. Although pathogen survival in water microcosms and manure-amended soils is well documented, little is known about...

  3. Fecal pollution source tracking toolbox for identification, evaluation and characterization of fecal contamination in receiving urban surface waters and groundwater.

    PubMed

    Tran, Ngoc Han; Gin, Karina Yew-Hoong; Ngo, Huu Hao

    2015-12-15

    The quality of surface waters/groundwater of a geographical region can be affected by anthropogenic activities, land use patterns and fecal pollution sources from humans and animals. Therefore, the development of an efficient fecal pollution source tracking toolbox for identifying the origin of the fecal pollution sources in surface waters/groundwater is especially helpful for improving management efforts and remediation actions of water resources in a more cost-effective and efficient manner. This review summarizes the updated knowledge on the use of fecal pollution source tracking markers for detecting, evaluating and characterizing fecal pollution sources in receiving surface waters and groundwater. The suitability of using chemical markers (i.e. fecal sterols, fluorescent whitening agents, pharmaceuticals and personal care products, and artificial sweeteners) and/or microbial markers (e.g. F+RNA coliphages, enteric viruses, and host-specific anaerobic bacterial 16S rDNA genetic markers) for tracking fecal pollution sources in receiving water bodies is discussed. In addition, this review also provides a comprehensive approach, which is based on the detection ratios (DR), detection frequencies (DF), and fate of potential microbial and chemical markers. DR and DF are considered as the key criteria for selecting appropriate markers for identifying and evaluating the impacts of fecal contamination in surface waters/groundwater.

  4. A coupled modeling and molecular biology approach to microbial source tracking at Cowell Beach, Santa Cruz, CA, United States.

    PubMed

    Russell, Todd L; Sassoubre, Lauren M; Wang, Dan; Masuda, Shelly; Chen, Helen; Soetjipto, Cherrie; Hassaballah, Abdulrahman; Boehm, Alexandria B

    2013-09-17

    Consistently high levels of bacterial indicators of fecal pollution rank Cowell Beach as the most polluted beach in California. High levels of fecal indicator bacteria (FIB), E. coli and enterococci, are measured throughout the summer, resulting in beach advisories with social and economic consequences. The source of FIB, however, is unknown. Speculations have been made that the wrack accumulating on the beach is a major source of FIB to the surf zone. The present study uses spatial and temporal sampling coupled with process-modeling to investigate potential FIB sources and the relative contributions of those sources. Temporal sampling showed consistently high FIB concentrations in the surf zone, sand, and wrack at Cowell Beach, and ruled out the storm drain, the river, the harbor, and the adjacent wharf as the sources of the high concentrations observed in the surf zone. Spatial sampling confirmed that the source of FIB to the beach is terrestrial rather than marine. Modeling results showed two dominant FIB sources to the surf zone: sand for enterococci and groundwater for E. coli. FIB from wrack represented a minor contribution to bacterial levels in the water. Molecular source tracking methods indicate the FIB at the beach is of human and bird origin. The microbial source tracking (MST) approach presented here provides a framework for future efforts.

  5. Oxygen isotope and salinity relationships from SE Alaska track freshwater sources in the NE Pacific

    NASA Astrophysics Data System (ADS)

    Mix, A. C.; Prahl, F. G.; Bruce, F.

    2006-12-01

    Water column d18O and salinity data (n=330) from hydrocasts at 31 sites occupied off southeast Alaska in August and September of 2004 constrain regional freshwater sources to the ocean. At open-ocean sites, d18O at the zero-salinity intercept (DF) decreases systematically from near the Canadian border (55 degN, DF = -14 permil VSMOW) to Hinchinbrook Entrance (~61 degN, DF = -18 permil VSMOW), and closely tracks estimates of d18O of local precipitation at sealevel (based on the OICP database, perhaps with significant contributions from the Copper River in the north). A similar result is obtained for fjord sites, suggesting relatively little contribution of melting glacier ice. This finding includes sites where floating ice is clearly melting, such as Yakutat and Disenchantment Bays (~60 degN). Here, DF is -18 permil VSMOW, consistent with regional sealevel precipitation (-17 to -18 permil, OICP database), and not consistent with substantial contributions from ice (melting bergs, likely from Hubbard Glacier were measured with d18O of -25.7 permil VSMOW). An implication is that the freshwater budget in the marine system off SE Alaska is dominated today by local precipitation, rather than melting ice, and that regional paleoceanographic estimates of paleosalinity based on stable isotope data will primarily reflect this process.

  6. Relationship of human-associated microbial source tracking markers with Enterococci in Gulf of Mexico waters.

    PubMed

    Gordon, Katrina V; Brownell, Miriam; Wang, Shiao Y; Lepo, Joe Eugene; Mott, Joanna; Nathaniel, Rajkumar; Kilgen, Marilyn; Hellein, Kristen N; Kennedy, Elizabeth; Harwood, Valerie J

    2013-03-01

    Human and ecosystem health can be damaged by fecal contamination of recreational waters. Microbial source tracking (MST) can be used to specifically detect domestic sewage containing human waste, thereby informing both risk assessment and remediation strategies. Previously, an inter-laboratory collaboration developed standardized PCR methods for a bacterial, an archaeal, and a viral indicator of human sewage. Here we present results for two subsequent years of field testing in fresh and salt water by five laboratories across the U.S. Gulf Coast (two in Florida and one each in Mississippi, Louisiana and Texas) using common standard operating procedures (SOPs) developed previously. Culturable enterococci were enumerated by membrane filtration, and PCR was used to detect three MST markers targeting domestic sewage: human-associated Bacteroides (HF183), Methanobrevibacter smithii and human polyomaviruses BK and JC (HPyVs). Detection of sewage markers in surface waters was significantly associated with higher enterococci levels and with exceedance of the recreational water quality standard in four or three regions, respectively. Sewage markers were frequently co-detected in single samples, e.g., M. smithii and HF183 were co-detected in 81% of Louisiana samples, and HPyVs and M. smithii were co-detected in over 40% of southwest Florida and Mississippi samples. This study demonstrates the robustness and inter-laboratory transferability of these three markers for the detection of pollution from domestic sewage in the waters impacting the Gulf of Mexico over a coastal range of over 1000 miles.

  7. Occurrence of Cryptosporidium oocysts and Giardia cysts in effluent from sewage treatment plant from eastern Poland.

    PubMed

    Sroka, Jacek; Stojecki, Krzysztof; Zdybel, Jolanta; Karamon, Jacek; Cencek, Tomasz; Dutkiewicz, Jacek

    2013-01-01

    Cryptosporidium spp. and Giardia lamblia (synonyms: Giardia duodenalis, Giardia intestinalis) are emerging protozoa causing disease in humans and animals worldwide. These parasites can pose a serious threat to immunocompromised people, for whom the symptoms are more severe and may include abdominal pain, watery diarrhoea, nausea, headaches, malaise, and fever. One of the sources of these parasites can be treated wastewater from wastewater treatment plants (WTPs). Samples of treated wastewater (effluent), each of 10 L volume, were collected from 13 municipal WTPs located in eastern Poland. Cryptosporidium oocysts and Giardia cysts were separated by the immunomagnetic method. The presence and/or concentration of protozoan (oo)cysts in effluent samples were determined by direct immunofluorescent microscopy, nested PCR and Real Time PCR. Viability of (oo)cysts was determined by double-staining with the use of Live/Dead BacLight kit (Invitrogen). Cryptosporidium spp. oocysts were detected in 8 WTPs (61.5%) and Giardia spp. cysts in 11 WTPs (84.6%) by microscopic analysis. Both pathogens were detected in samples from 7 WTPs. Median concentrations of Cryptosporidium and Giardia (oo)cysts in 13 examined samples were 2.2/L and 6.6/L, respectively, while mean concentrations were 28.5/L and 113.6/L, respectively. In positive samples, Cryptosporidium oocysts concentrations ranged from 0.4 - 154.1 oocysts per litre, and Giardia cysts concentrations ranged from 0.7 - 660 cysts per litre. By nested PCR, Giardia DNA was detected in 4 samples of the 13 examined, (30.8%) while Cryptosporidium DNA was never detected. In Real Time PCR, positive results for Giardia were obtained in 5 samples (38.5%) and in none of the samples for Cryptosporidium, with the exception of one equivocal result. Viable (oo)cysts of Cryptosporidium and Giardia were detected in 3 out of 4 samples examined, in the ranges of 12.5 - 60% and 50 - 100% of total (oo)cysts, respectively. In view of our preliminary

  8. Variability in susceptibility of voles (Arvicolinae) to experimental infection with Cryptosporidium muris and Cryptosporidium andersoni.

    PubMed

    Modrý, David; Hofmannová, Lada; Antalová, Zuzana; Sak, Bohumil; Kváč, Martin

    2012-07-01

    The infectivity of Cryptosporidium muris and Cryptosporidium andersoni in various species of voles was studied using experimental infections. None of the experimental voles inoculated with 1 × 10(5) oocysts of Cryptosporidium spp. shed any oocysts during 40 DPI, except Brandt's vole (Lasiopodomys brandtii), which was susceptible to C. muris infection. Experiments confirmed the resistance of voles of the genus Microtus sensu stricto to infection with mammalian gastric cryptosporidia, which provides a new study model with prospects to more fully understand the processes involved in the phenomenon of host specificity of this group of protists.

  9. Cost-effective method for microbial source tracking using specific human and animal viruses.

    PubMed

    Bofill-Mas, Silvia; Hundesa, Ayalkibet; Calgua, Byron; Rusiñol, Marta; Maluquer de Motes, Carlos; Girones, Rosina

    2011-12-03

    Microbial contamination of the environment represents a significant health risk. Classical bacterial fecal indicators have shown to have significant limitations, viruses are more resistant to many inactivation processes and standard fecal indicators do not inform on the source of contamination. The development of cost-effective methods for the concentration of viruses from water and molecular assays facilitates the applicability of viruses as indicators of fecal contamination and as microbial source tracking (MST) tools. Adenoviruses and polyomaviruses are DNA viruses infecting specific vertebrate species including humans and are persistently excreted in feces and/or urine in all geographical areas studied. In previous studies, we suggested the quantification of human adenoviruses (HAdV) and JC polyomaviruses (JCPyV) by quantitative PCR (qPCR) as an index of human fecal contamination. Recently, we have developed qPCR assays for the specific quantification of porcine adenoviruses (PAdV) and bovine polyomaviruses (BPyV) as animal fecal markers of contamination with sensitivities of 1-10 genome copies per test tube. In this study, we present the procedure to be followed to identify the source of contamination in water samples using these tools. As example of representative results, analysis of viruses in ground water presenting high levels of nitrates is shown. Detection of viruses in low or moderately polluted waters requires the concentration of the viruses from at least several liters of water into a much smaller volume, a procedure that usually includes two concentration steps in series. This somewhat cumbersome procedure and the variability observed in viral recoveries significantly hamper the simultaneous processing of a large number of water samples. In order to eliminate the bottleneck caused by the two-step procedures we have applied a one-step protocol developed in previous studies and applicable to a diversity of water matrices. The procedure includes

  10. Cost-effective Method for Microbial Source Tracking Using Specific Human and Animal Viruses

    PubMed Central

    Bofill-Mas, Sílvia; Hundesa, Ayalkibet; Calgua, Byron; Rusiñol, Marta; Maluquer de Motes, Carlos; Girones, Rosina

    2011-01-01

    Microbial contamination of the environment represents a significant health risk. Classical bacterial fecal indicators have shown to have significant limitations, viruses are more resistant to many inactivation processes and standard fecal indicators do not inform on the source of contamination. The development of cost-effective methods for the concentration of viruses from water and molecular assays facilitates the applicability of viruses as indicators of fecal contamination and as microbial source tracking (MST) tools. Adenoviruses and polyomaviruses are DNA viruses infecting specific vertebrate species including humans and are persistently excreted in feces and/or urine in all geographical areas studied. In previous studies, we suggested the quantification of human adenoviruses (HAdV) and JC polyomaviruses (JCPyV) by quantitative PCR (qPCR) as an index of human fecal contamination. Recently, we have developed qPCR assays for the specific quantification of porcine adenoviruses (PAdV) and bovine polyomaviruses (BPyV) as animal fecal markers of contamination with sensitivities of 1-10 genome copies per test tube. In this study, we present the procedure to be followed to identify the source of contamination in water samples using these tools. As example of representative results, analysis of viruses in ground water presenting high levels of nitrates is shown. Detection of viruses in low or moderately polluted waters requires the concentration of the viruses from at least several liters of water into a much smaller volume, a procedure that usually includes two concentration steps in series. This somewhat cumbersome procedure and the variability observed in viral recoveries significantly hamper the simultaneous processing of a large number of water samples. In order to eliminate the bottleneck caused by the two-step procedures we have applied a one-step protocol developed in previous studies and applicable to a diversity of water matrices. The procedure includes

  11. Cryptosporidium infections: a laboratory survey.

    PubMed Central

    Elsser, K A; Moricz, M; Proctor, E M

    1986-01-01

    Between Oct. 1, 1983, and June 30, 1985, Cryptosporidium oocysts were identified in stool specimens from 74 patients who presented with gastrointestinal symptoms to their physicians. Questionnaires prepared to determine travel history, symptoms, duration of illness and epidemiologic characteristics of the infection were completed for 67 (90%) of the patients by their physicians; the information on the other 7 patients was obtained from the requisitions accompanying the specimens. Of the 67, 35 (52%) had recently been to Mexico. The infection was likely transmitted through contaminated water, food and, possibly, milk. The infections in patients who had not travelled were thought to be due to contact with infected pets or farm animals or with infected children attending daycare centres. Diarrhea, vomiting, fever and nausea usually lasted for 1 to 2 weeks, except in those with immune deficiency, in whom the symptoms persisted for up to 6 months. The condition was diagnosed by identification of oocysts in stool specimens that underwent formalin-ether sedimentation and modified cold Kinyoun staining. PMID:3730980

  12. Detection of Cryptosporidium and Giardia in agricultural and water environments in the Qinghai area of China by IFT and PCR.

    PubMed

    Ma, Liqing; Sotiriadou, Isaia; Cai, Qigang; Karanis, Gabriele; Wang, Geping; Wang, Guanghua; Lu, Yan; Li, Xiuping; Karanis, Panagiotis

    2014-09-01

    Qinghai Province in northwest China is strongly influenced by agricultural activities and is an important source of food and drinking water. Here, we present findings regarding the occurrence and molecular epidemiology of Cryptosporidium and Giardia species based on a large-scale investigation of areas of Qinghai Province. The diagnosis and molecular detection of Cryptosporidium oocysts and Giardia cysts was carried out using immunofluorescence microscopy (IFT), whereas nested polymerase chain reaction (PCR) in fecal smears and water samples was used for the detection and molecular characterization of the species. In total, 561 samples (260 water samples and 301 fecal samples from animals) were collected and analyzed. Of the 260 water samples, 66 samples were Cryptosporidium-positive by IFT and 71 samples were positive by nested PCR; in addition, 39 samples were Giardia-positive by IFT and 40 samples were positive by nested PCR. Of the 301 fecal samples from animals, 98 samples were Cryptosporidium-positive by IFT and 61 samples were positive by nested PCR, whereas 52 samples were Giardia-positive by IFT and 31 samples were positive by nested PCR. We showed that the water supplies and animals investigated contained Cryptosporidium and Giardia (oo)cysts. Thus, we recommend that the Chinese Government and Chinese health authorities undertake control measures to protect the food and drinking water sources in Qinghai from these pathogenic protozoa.

  13. Pathogenic Mechanisms of Cryptosporidium and Giardia.

    PubMed

    Certad, Gabriela; Viscogliosi, Eric; Chabé, Magali; Cacciò, Simone M

    2017-03-20

    Intestinal protozoa are important etiological agents of diarrhea, particularly in children, yet the public health risk they pose is often neglected. Results from the Global Enteric Multicenter Study (GEMS) showed that Cryptosporidium is among the leading causes of moderate to severe diarrhea in children under 2 years. Likewise, Giardia infects approximately 200 million individuals worldwide, and causes acute diarrhea in children under 5 years. Despite this recognized role as pathogens, the question is why and how these parasites cause disease in some individuals but not in others. This review focuses on known pathogenic mechanisms of Cryptosporidium and Giardia, and infection progress towards disease.

  14. 40 CFR 141.716 - Source toolbox components.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... undertake to reduce source water Cryptosporidium levels. The plan must explain how the actions are expected... will reduce source water Cryptosporidium levels. (4) If the State does not respond to a system.... If any change is likely to reduce the level of source water protection, the system must also list...

  15. 40 CFR 141.716 - Source toolbox components.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... undertake to reduce source water Cryptosporidium levels. The plan must explain how the actions are expected... will reduce source water Cryptosporidium levels. (4) If the State does not respond to a system.... If any change is likely to reduce the level of source water protection, the system must also list...

  16. Molecular identification of Enterocytozoon bieneusi, Cryptosporidium, and Giardia in Brazilian captive birds.

    PubMed

    da Cunha, Maria Júlia Rodrigues; Cury, Márcia Cristina; Santín, Mónica

    2017-02-01

    A total of 85 fecal samples from captive birds collected from October 2013 to September 2014 in Uberlândia and Belo Horizonte in the state of Minas Gerais (Brazil) were evaluated for the presence of Enterocytozoon bieneusi, Cryptosporidium, and Giardia by PCR. Of these, three birds were found positive for E. bieneusi (3.5%), two for Cryptosporidium (2.3%), and one for Giardia (1.2%). Two genotypes of E. bieneusi were detected by nucleotide sequence analysis of the ITS region, genotypes D and Peru 6 in a swan goose and in two rock pigeons, respectively. For Cryptosporidium and Giardia, nucleotide sequence analysis of the SSU rRNA identified Cryptosporidium baileyi and Duck genotype in a swan goose and a mandarin duck, respectively, and Giardia duodenalis assemblage A in a toco toucon. Our results demonstrate that human-pathogenic E. bieneusi genotypes D and Peru6 and G. duodenalis assemblage A are present in captive birds in Brazil, corroborating their potential role as a source of human infection and environmental contamination.

  17. Occurrence and molecular characterization of Cryptosporidium spp. genotypes in European hedgehogs (Erinaceus europaeus L.) in Germany.

    PubMed

    Dyachenko, V; Kuhnert, Y; Schmaeschke, R; Etzold, M; Pantchev, N; Daugschies, A

    2010-02-01

    Juvenile hedgehogs having insufficient body weight are often brought for overwintering to hedgehog rehabilitation centres. Faecal samples of juvenile hedgehogs and overwintering hedgehogs (n=188) collected prior to releasing them back into the wilderness were examined for the presence of Cryptosporidium coproantigen and oocysts. Altogether 56 (29.8%) submitted samples were positive for coproantigen. Forty-five (39.5%, n=114) of the positive samples originated from newly rescued hedgehogs, while 11 (14.8%, n=74) positive samples were from animals that spent several months at the station. Fifteen samples subjected to PCR-RFLP analysis on the partial 18S rRNA locus suggested the presence of C. parvum. Multilocus sequence typing on partial 60 kDa glycoprotein gene, 18S rRNA, actin gene, 70 kDa heat shock protein gene sequences revealed 3 different subtype families: IIa, IIc and a new, proposed as VIIa subtype family. Cryptosporidium sp. genotype belonging to VIIa subtype family is closely related to C. parvum but is genetically distinct being probably a hedgehog-specific Cryptosporidium sp. genotype with unknown zoonotical potential. Hedgehogs excreting Cryptosporidium oocysts represent a potential source for human infections, but also an anthroponotic nature of the IIc subtype family should be reviewed.

  18. Cryptosporidium parvum scavenges LDL-derived cholesterol and micellar cholesterol internalized into enterocytes

    PubMed Central

    Ehrenman, Karen; Wanyiri, Jane W.; Bhat, Najma; Ward, Honorine D.; Coppens, Isabelle

    2013-01-01

    Cryptosporidium spp. are responsible for devastating diarrhea in immunodeficient individuals. In the intestinal tract, the developmental stages of the parasite are confined to the apical surfaces of epithelial cells. Upon invasion, Cryptosporidium incorporates the microvillous membrane of the enterocyte to form the parasitophorous vacuole (PV) and sequesters itself from the host cytoplasm by rearranging the host cytoskeleton. Cryptosporidium parvum has minimal anabolic capabilities and relies on transporters and salvage pathways to meet its basic metabolic requirements. The cholesterol salvage pathway is crucial for the development of protozoan parasites. In this study, we have examined the sources of cholesterol from C. parvum infecting enterocytes. We illustrated that the intracellular stages of Cryptosporidium as well as the oocysts shed by the host, contain cholesterol. Incubation of infected enterocytes in lipoprotein-free medium impairs parasite development and results in substantial decrease in cholesterol content associated with the PV. Among lipoproteins, LDL constitutes an important source of cholesterol for Cryptosporidium. Dietary cholesterol incorporated into micelles is internalized into enterocytes by the NPC1L1 transporter. We showed that C. parvum also obtains cholesterol from micelles in enterocytes. Pharmacological blockade of NPC1L1 function by ezetimibe or moderate down-regulation of NPC1L1 expression decreases parasite infectivity. These observations indicate that, despite its dual sequestration from the intestinal lumen and the host cytoplasm, C. parvum can, in fact, obtain cholesterol both from the gut’s lumen and the host cell. This study highlights the evolutionary advantages for epicellular pathogens to access to nutrients from the outside and inside of the host cell. PMID:23311949

  19. Cryptosporidium suis and Cryptosporidium scrofarum in Eurasian wild boars (Sus scrofa) in Central Europe.

    PubMed

    Němejc, Karel; Sak, Bohumil; Květoňová, Dana; Hanzal, Vladimír; Janiszewski, Paweł; Forejtek, Pavel; Rajský, Dušan; Ravaszová, Petra; McEvoy, John; Kváč, Martin

    2013-11-08

    From 2011 to 2012, to identify Cryptosporidium spp. occurrence in Eurasian wild boars (Sus scrofa) 29 randomly selected localities (both forest areas and enclosures) across the Central European countries of Austria, the Czech Republic, Poland, and the Slovak Republic were investigated. Cryptosporidium oocysts were microscopicaly detected in 11 out of 460 faecal samples examined using aniline-carbol-methyl violet staining. Sixty-one Cryptosporidium infections, including the 11 infections that were detected by microscopy, were detected using genus- or species-specific nested PCR amplification of SSU rDNA. This represents a 5.5 fold greater sensitivity for PCR relative to microscopy. Combining genus- and species-specific PCR tools significantly changes the perspective on the occurrence of Cryptosporidium spp. in wild boars. While RFLP and direct sequencing of genus specific PCR-amplified products revealed 56 C. suis (20) and C. scrofarum (36) monoinfections and only 5 mixed infections of these species, species-specific molecular tools showed 44 monoinfections and 17 mixed infections with these species. PCR analysis of the gp60 gene did not reveal any other Cryptosporidium infections. Similar to domestic pigs, C. scrofarum was detected as a dominant species infecting adult Eurasian wild boars (Sus scrofa). Cryptosporidium infected wild boars did not show signs of clinical disease. This report is perhaps the most comprehensive survey of cryptosporidial infection in wild boars.

  20. Utility of Microbial Source-Tracking Markers for Assessing Fecal Contamination in the Portage River Watershed, Northwestern Ohio, 2008

    USGS Publications Warehouse

    Kephart, Christopher M.; Bushon, Rebecca N.

    2010-01-01

    An influx of concentrated animal feeding operations in northwest Ohio has prompted local agencies to examine the effects of these industrial farms on water quality in the upper Portage River watershed. The utility of microbial source-tracking (MST) tools as a means of characterizing sources of fecal contamination in the watershed was evaluated. From 2007 to 2008, scientists with the U.S. Geological Survey, Bowling Green State University, and the Wood County Health Department collected and analyzed 17 environmental samples and 13 fecal source samples for Bacteroides-based host-associated DNA markers. At many of the environmental sites tested, MST marker results corroborated the presumptive fecal contamination sources. Results from this demonstration study support the utility of using MST with host-specific molecular markers to characterize the sources of fecal contamination in the Portage River watershed.

  1. A new high-resolution central and western Saharan summertime dust source map from automated satellite dust plume tracking

    NASA Astrophysics Data System (ADS)

    Ashpole, Ian; Washington, Richard

    2013-07-01

    In this paper, we outline a new objective dust source detection method for the central and western Sahara (CWS), based on the automated tracking of individual dust plumes in data from the Spinning Enhanced Visible and Infrared Imager, available every 15 mins. at ~0.03° spatial resolution. The method is used to map the origin of summertime dust storms in the CWS for June - August 2004 - 2010. It reveals the sources of these events in unprecedented detail, allowing for the identification of specific, highly active source areas. The study of collocated surface features reveals that many of the dominant sources are likely associated with paleolakes and outwash plains, many in close proximity to the Saharan mountains. Extensive nonsource areas are associated with low albedo and elevated terrain, pointing to the mountainous regions of the Sahara. Additionally, sand seas are not identified as important source areas, but their margins sometimes are. The automated tracking method also facilitates analysis of the transport direction of dust plumes from key source regions and the inference of emission mechanisms. It is found that there are two broad domains within the CWS: one in southwest Algeria and northwest Mali, characterized primarily by transport toward the southwest and very likely dominated by low-level jets embedded in the northeasterly Harmattan winds; and a second in southern Algeria, northwest Niger, and northeast Mali where there is no preferred transport direction and a strong potential association between dust events and deep convection, pointing toward cold pool outflows as the likely deflation mechanism.

  2. Marine Animal Sound Database. Twelve Years of Tracking 52-Hz Whale Calls from a Unique Source in the North Pacific

    DTIC Science & Technology

    2007-11-02

    THIS PAGE ABSTRACT OF William A. Watkins PAGESUnclass Unclass Unclass UL 19b. TELEPHONE NUMBER Ilnclude area code) (508) 289-3258 (Mary Ann Daher ...ABSTRACTS: In Press: Watkins, W.A., Daher , M.A. and George, J.E. 2004. Twelve years of tracking 52-Hz whale calls from a unique source in the North...Watkins, Mary Ann Daher *, Joseph E. George, David Rodriguez Woodv Hole Oceanographic Institution. MS 36, Woodv Hole, MA 02543, USA Received 2 March 2004

  3. Cryptosporidium Oocysts in a Water Supply Associated with a Cryptosporidiosis Outbreak

    PubMed Central

    Howe, Andrew D.; Forster, Sue; Morton, Stephen; Marshall, Roberta; Osborn, Keith S.; Wright, Peter

    2002-01-01

    An outbreak of cryptosporidiosis occurred in and around Clitheroe, Lancashire, in northwest England, during March 2000. Fifty-eight cases of diarrhea with Cryptosporidium identified in stool specimens were reported. Cryptosporidium oocysts were identified in samples from the water treatment works as well as domestic taps. Descriptive epidemiology suggested that drinking unboiled tap water in a single water zone was the common factor linking cases. Environmental investigation suggested that contamination with animal feces was the likely source of the outbreak. This outbreak was unusual in that hydrodynamic modeling was used to give a good estimate of the peak oocyst count at the time of the contamination incident. The oocysts’ persistence in the water distribution system after switching to another water source was also unusual. This persistence may have been due to oocysts being entrapped within biofilm. Despite the continued presence of oocysts, epidemiologic evidence suggested that no one became ill after the water source was changed. PMID:12023920

  4. Molecular Genotyping of Viable Cryptosporidium Oocysts

    EPA Science Inventory

    Cryptosporidium is a chlorination-resistant protozoan parasite that causes a self-limiting diarrheal disease in the immunocompetent or severe chronic diarrhea in the immunocompromised. Two species, C. parvum and C. hominis, cause most cases of cryptosporidiosis in humans, while C...

  5. RELIABILITY OF SURROGATES FOR DETERMINING CRYPTOSPORIDIUM REMOVAL

    EPA Science Inventory

    Testing of field-scale bag filtration systems yielded results indicating that 4-6-um polystyrene microspheres can be used as a reliable surrogate for determining Cryptosporidium oocyst removal in bag filtration process. A nearly perfect linear correlation was observed between log...

  6. Identification of Cryptosporidium oocysts in river water.

    PubMed Central

    Ongerth, J E; Stibbs, H H

    1987-01-01

    Water samples were collected from four rivers in Washington State and two rivers in California and examined for the presence of Cryptosporidium oocysts. Oocyst-sized particles were concentrated from 20-liter samples of water by membrane filtration, centrifugation, and differential sedimentation. The particle concentrate was then deposited on a 25-mm-diameter membrane filter for oocyst identification by indirect immunofluorescence assay. The identification procedure had a limit of detection of about five oocysts per liter. Cryptosporidium oocysts were found in each of 11 river water samples examined. Concentrations ranged from 2 to 112 oocysts per liter. The finding of Cryptosporidium oocysts in all samples examined from six western rivers is noteworthy in light of recent reports indicating that Cryptosporidium sp. is a significant agent of human and animal disease. This finding suggests that waterborne oocysts of this parasite are more important than was previously recognized. More detailed studies are needed to define geographical and temporal distribution, to assess the viability of waterborne oocysts, and to determine the importance of water as a means of transmission. Images PMID:3579275

  7. 10 CFR 20.2207 - Reports of transactions involving nationally tracked sources.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... radioactive material in the source; (5) The initial source strength in becquerels (curies) at the time of... the source; (6) The initial or current source strength in becquerels (curies); (7) The date for which... in becquerels (curies); (7) The date for which the source strength is reported; (8) The date...

  8. 10 CFR 20.2207 - Reports of transactions involving nationally tracked sources.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... radioactive material in the source; (5) The initial source strength in becquerels (curies) at the time of... the source; (6) The initial or current source strength in becquerels (curies); (7) The date for which... in becquerels (curies); (7) The date for which the source strength is reported; (8) The date...

  9. 10 CFR 20.2207 - Reports of transactions involving nationally tracked sources.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... radioactive material in the source; (5) The initial source strength in becquerels (curies) at the time of... the source; (6) The initial or current source strength in becquerels (curies); (7) The date for which... in becquerels (curies); (7) The date for which the source strength is reported; (8) The date...

  10. 10 CFR 20.2207 - Reports of transactions involving nationally tracked sources.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... radioactive material in the source; (5) The initial source strength in becquerels (curies) at the time of... the source; (6) The initial or current source strength in becquerels (curies); (7) The date for which... in becquerels (curies); (7) The date for which the source strength is reported; (8) The date...

  11. 10 CFR 20.2207 - Reports of transactions involving nationally tracked sources.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... radioactive material in the source; (5) The initial source strength in becquerels (curies) at the time of... the source; (6) The initial or current source strength in becquerels (curies); (7) The date for which... in becquerels (curies); (7) The date for which the source strength is reported; (8) The date...

  12. Can antibiotic resistance analysis be a useful tool for tracking population sources of contamination in Yucatan groundwater?

    NASA Astrophysics Data System (ADS)

    McLain, J. E.; Lenczewski, M.; Leal-Bautista, R. M.

    2013-05-01

    Antibiotic resistance patterns have been widely used in scientific studies conducted to identify sources of water contamination. However, the methods of resistance determination have not been standardized; therefore, the data on antibiotic resistance in the environment come from studies that have used a range of media types, antibiotic concentrations, and incubation periods, making it difficult to compare results between environments. Over two years, we assessed antibiotic sensitivity of Gram-positive and Gram-negative bacteria isolated from Yucatan water sources, to identify unique resistance patterns and assess the potential for antibiotic resistance analysis as a tool to discriminate between fecal pollution from two population sources (tourist and local). Though resistance to erythromycin, streptomycin, and ciprofloxacin showed some differences between populations, natural bacterial resistance (in isolates from pristine sources) was very high and confounded the research findings. This study highlights the need among the research community involved in tracking of environmental antibiotic resistance to develop a standardized and rigorously validated suite of methods that address background resistance and that can be used across environments, to accurately inform source tracking studies.

  13. The first report on Cryptosporidium suis and Cryptosporidium pig genotype II in Eurasian wild boars (Sus scrofa) (Czech Republic).

    PubMed

    Němejc, Karel; Sak, Bohumil; Květoňová, Dana; Hanzal, Vladimír; Jeníková, Martina; Kváč, Martin

    2012-03-23

    A total of 193 faecal samples of adult Eurasian wild boars were collected at 12 enclosures across the Czech Republic and examined for Cryptosporidium infection using both microscopic and molecular tools. Cryptosporidium oocysts were not detected in any of the 193 faecal samples examined using the aniline-carbol-methyl violet staining method. Thirty-two positive cases of Cryptosporidium infection were detected using either genus- or species-specific nested PCR. Mono-infection with Cryptosporidium suis and Cryptosporidium pig genotype II were found in 13 and 7 cases, respectively. Five mixed infections of C. suis and Cryptosporidium pig genotype II were detected using PCR/RFLP with genus specific primers. The number of detected mixed infections increased 2.4 fold when a species-specific PCR was employed. No other Cryptosporidium spp. was detected. Unlike cryptosporidiosis of domestic pigs, C. suis was detected as a dominant species infecting adult Eurasian wild boars. There was no association between diarrhoea and the presence of Cryptosporidium infection in the Eurasian wild boars studied. This is the first report on the Cryptosporidium infection caused by C. suis and Cryptosporidium pig genotype II in Eurasian wild boars (Sus scrofa).

  14. Household Socioeconomic and Demographic Correlates of Cryptosporidium Seropositivity in the United States

    PubMed Central

    Becker, Daniel J.; Oloya, James; Ezeamama, Amara E.

    2015-01-01

    Background Cryptosporidium are parasitic protozoa that infect humans, domestic animals, and wildlife globally. In the United States, cryptosporidiosis occurs in an estimated 750,000 persons annually, and is primarily caused by either of the Cryptosporidium parvum genotypes 1 and 2, exposure to which occurs through ingestion of food or water contaminated with oocytes shed from infected hosts. Although most cryptosporidiosis cases are caused by genotype 1 and are of human origin, the zoonotic sources of genotype 2, such as livestock, are increasingly recognized as important for understanding human disease patterns. Social inequality could mediate patterns of human exposure and infection by placing individuals in environments where food or water contamination and livestock contact is high or through reducing the availability of educational and sanitary resources required to avoid exposure. Methodology/Principal Findings We here analyzed data from the National Health and Nutritional Examination Survey (NHANES) between 1999 and 2000, and related seropositivity to Cryptosporidium parvum to correlates of social inequality at the household and individual scale. After accounting for the complex sampling design of NHANES and confounding by individual demographics and household conditions, we found impaired household food adequacy was associated with greater odds of Cryptosporidium seropositivity. Additionally, we identified individuals of non-white race and ethnicity and those born outside the United States as having significantly greater risk than white, domestic-born counterparts. Furthermore, we provide suggestive evidence for direct effects of family wealth on Cryptosporidium seropositivity, in that persons from low-income households and from families close to the poverty threshold had elevated odds of seropositivity relative to those in high-income families and in households far above the poverty line. Conclusions/Significance These results refute assertions that

  15. Comparing passive source localization and tracking approaches with a towed horizontal receiver array in an ocean waveguide.

    PubMed

    Gong, Zheng; Tran, Duong D; Ratilal, Purnima

    2013-11-01

    Approaches for instantaneous passive source localization using a towed horizontal receiver array in a random range-dependent ocean waveguide are examined. They include: (1) Moving array triangulation, (2) array invariant, (3) bearings-only target motion analysis in modified polar coordinates via the extended Kalman filter, and (4) bearings-migration minimum mean-square error. These methods are applied to localize and track a vertical source array deployed in the far-field of a towed horizontal receiver array during the Gulf of Maine 2006 Experiment. The source transmitted intermittent broadband pulses in the 300 to 1200 Hz frequency range. A nonlinear matched-filter kernel designed to replicate the acoustic signal measured by the receiver array is applied to enhance the signal-to-noise ratio. The source localization accuracy is found to be highly dependent on source-receiver geometry and the localization approach. For a relatively stationary source drifting at speeds much slower than the receiver array tow-speed, the mean source position can be estimated by moving array triangulation with less than 3% error near broadside direction. For a moving source, the Kalman filter method gives the best performance with 5.5% error. The array invariant is the best approach for localizing sources within the endfire beam of the receiver array with 7% error.

  16. Noncoherent Tracking of a Source of a Data-Modulated Signal

    NASA Technical Reports Server (NTRS)

    Arndt, G. Dickey; Ngo, Phong; Chen, Henry; Phan, Chau T.; Hill, Brent; Bourgeois, Brian; Dusl, John

    2003-01-01

    A proposed tracking receiver system containing three suitably positioned antenna elements and special signal-processing equipment would determine the direction of incidence of a microwave signal containing spread-spectrum digital data modulation. If the system were to contain two sets of antenna elements separated by a known baseline, it could determine the location of the transmitter as the intersection of the lines of incidence on the two antennas. Such systems could be used for diverse purposes in outer space and on Earth, including tracking astronauts and small robotic spacecraft working outside a spacecraft or space station, and locating cellular telephones from which distress calls have been made. The principle of operation does not require the transmission of a special identifying or distress signal by the cellular telephone or other transmitter to be tracked; instead, the system could utilize the data signal routinely sent by the transmitter, provided that the signal had the characteristics needed for processing.

  17. Dual wavelength multiple-angle light scattering system for cryptosporidium detection

    NASA Astrophysics Data System (ADS)

    Buaprathoom, S.; Pedley, S.; Sweeney, S. J.

    2012-06-01

    A simple, dual wavelength, multiple-angle, light scattering system has been developed for detecting cryptosporidium suspended in water. Cryptosporidium is a coccidial protozoan parasite causing cryptosporidiosis; a diarrheal disease of varying severity. The parasite is transmitted by ingestion of contaminated water, particularly drinking-water, but also accidental ingestion of bathing-water, including swimming pools. It is therefore important to be able to detect these parasites quickly, so that remedial action can be taken to reduce the risk of infection. The proposed system combines multiple-angle scattering detection of a single and two wavelengths, to collect relative wavelength angle-resolved scattering phase functions from tested suspension, and multivariate data analysis techniques to obtain characterizing information of samples under investigation. The system was designed to be simple, portable and inexpensive. It employs two diode lasers (violet InGaN-based and red AlGaInP-based) as light sources and silicon photodiodes as detectors and optical components, all of which are readily available. The measured scattering patterns using the dual wavelength system showed that the relative wavelength angle-resolved scattering pattern of cryptosporidium oocysts was significantly different from other particles (e.g. polystyrene latex sphere, E.coli). The single wavelength set up was applied for cryptosporidium oocysts'size and relative refractive index measurement and differential measurement of the concentration of cryptosporidium oocysts suspended in water and mixed polystyrene latex sphere suspension. The measurement results showed good agreement with the control reference values. These results indicate that the proposed method could potentially be applied to online detection in a water quality control system.

  18. Identification of Cryptosporidium Species in Fish from Lake Geneva (Lac Léman) in France.

    PubMed

    Certad, Gabriela; Dupouy-Camet, Jean; Gantois, Nausicaa; Hammouma-Ghelboun, Ourida; Pottier, Muriel; Guyot, Karine; Benamrouz, Sadia; Osman, Marwan; Delaire, Baptiste; Creusy, Colette; Viscogliosi, Eric; Dei-Cas, Eduardo; Aliouat-Denis, Cecile Marie; Follet, Jérôme

    2015-01-01

    Cryptosporidium, a protozoan parasite that can cause severe diarrhea in a wide range of vertebrates including humans, is increasingly recognized as a parasite of a diverse range of wildlife species. However, little data are available regarding the identification of Cryptosporidium species and genotypes in wild aquatic environments, and more particularly in edible freshwater fish. To evaluate the prevalence of Cryptosporidiumspp. in fish from Lake Geneva (Lac Léman) in France, 41 entire fish and 100 fillets (cuts of fish flesh) were collected from fishery suppliers around the lake. Nested PCR using degenerate primers followed by sequence analysis was used. Five fish species were identified as potential hosts of Cryptosporidium: Salvelinus alpinus, Esox lucius, Coregonus lavaretus, Perca fluviatilis, and Rutilus rutilus. The presence of Cryptosporidium spp. was found in 15 out of 41 fish (37%), distributed as follows: 13 (87%) C. parvum, 1 (7%) C. molnari, and 1 (7%) mixed infection (C. parvum and C. molnari). C. molnari was identified in the stomach, while C. parvum was found in the stomach and intestine. C. molnari was also detected in 1 out of 100 analyzed fillets. In order to identify Cryptosporidium subtypes, sequencing of the highly polymorphic 60-kDa glycoprotein (gp60) was performed. Among the C. parvum positive samples, three gp60 subtypes were identified: IIaA15G2R1, IIaA16G2R1, and IIaA17G2R1. Histological examination confirmed the presence of potential developmental stages of C. parvum within digestive epithelial cells. These observations suggest that C. parvum is infecting fish, rather than being passively carried. Since C. parvum is a zoonotic species, fish potentially contaminated by the same subtypes found in terrestrial mammals would be an additional source of infection for humans and animals, and may also contribute to the contamination of the environment with this parasite. Moreover, the risk of human transmission is strengthened by the

  19. Identification of Cryptosporidium Species in Fish from Lake Geneva (Lac Léman) in France

    PubMed Central

    Certad, Gabriela; Dupouy-Camet, Jean; Gantois, Nausicaa; Hammouma-Ghelboun, Ourida; Pottier, Muriel; Guyot, Karine; Benamrouz, Sadia; Osman, Marwan; Delaire, Baptiste; Creusy, Colette; Viscogliosi, Eric; Aliouat-Denis, Cecile Marie; Follet, Jérôme

    2015-01-01

    Cryptosporidium, a protozoan parasite that can cause severe diarrhea in a wide range of vertebrates including humans, is increasingly recognized as a parasite of a diverse range of wildlife species. However, little data are available regarding the identification of Cryptosporidium species and genotypes in wild aquatic environments, and more particularly in edible freshwater fish. To evaluate the prevalence of Cryptosporidiumspp. in fish from Lake Geneva (Lac Léman) in France, 41 entire fish and 100 fillets (cuts of fish flesh) were collected from fishery suppliers around the lake. Nested PCR using degenerate primers followed by sequence analysis was used. Five fish species were identified as potential hosts of Cryptosporidium: Salvelinus alpinus, Esox lucius, Coregonus lavaretus, Perca fluviatilis, and Rutilus rutilus. The presence of Cryptosporidium spp. was found in 15 out of 41 fish (37%), distributed as follows: 13 (87%) C. parvum, 1 (7%) C. molnari, and 1 (7%) mixed infection (C. parvum and C. molnari). C. molnari was identified in the stomach, while C. parvum was found in the stomach and intestine. C. molnari was also detected in 1 out of 100 analyzed fillets. In order to identify Cryptosporidium subtypes, sequencing of the highly polymorphic 60-kDa glycoprotein (gp60) was performed. Among the C. parvum positive samples, three gp60 subtypes were identified: IIaA15G2R1, IIaA16G2R1, and IIaA17G2R1. Histological examination confirmed the presence of potential developmental stages of C. parvum within digestive epithelial cells. These observations suggest that C. parvum is infecting fish, rather than being passively carried. Since C. parvum is a zoonotic species, fish potentially contaminated by the same subtypes found in terrestrial mammals would be an additional source of infection for humans and animals, and may also contribute to the contamination of the environment with this parasite. Moreover, the risk of human transmission is strengthened by the

  20. Prevalence and molecular characterization of Cryptosporidium in goats across four provincial level areas in China.

    PubMed

    Mi, Rongsheng; Wang, Xiaojuan; Huang, Yan; Zhou, Peng; Liu, Yuxuan; Chen, Yongjun; Chen, Jun; Zhu, Wei; Chen, Zhaoguo

    2014-01-01

    This study assessed the prevalence, species and subtypes of Cryptosporidium in goats from Guangdong Province, Hubei Province, Shandong Province, and Shanghai City of China. Six hundred and four fecal samples were collected from twelve goat farms, and the overall infection rate was 11.4% (69/604). Goats infected with Cryptosporidium were found in eleven farms across four provincial areas, and the infection rate ranged from 2.9% (1/35) to 25.0% (9/36). Three Cryptosporidium species were identified. Cryptosporidium xiaoi (45/69, 65.2%) was the dominant species, followed by C. parvum (14/69, 20.3%) and C. ubiquitum (10/69, 14.5%). The infection rate of Cryptosporidium spp. was varied with host age and goat kids were more susceptible to be infected than adult goats. Subtyping C. parvum and C. ubiquitum positive samples revealed C. parvum subtype IIdA19G1 and C. ubiquitum subtype XIIa were the most common subtypes. Other C. parvum subtypes were detected as well, such as IIaA14G2R1, IIaA15G1R1, IIaA15G2R1 and IIaA17G2R1. All of these subtypes have also been detected in humans, suggesting goats may be a potential source of zoonotic cryptosporidiosis. This was the first report of C. parvum subtypes IIaA14G2R1, IIaA15G1R1 and IIaA17G2R1 infecting in goats and the first molecular identification of C. parvum and its subtypes in Chinese goats.

  1. Serological responses to Cryptosporidium antigens in inhabitants of Hungary using conventionally filtered surface water and riverbank filtered drinking water.

    PubMed

    Farkas, K; Plutzer, J; Moltchanova, E; Török, A; Varró, M J; Domokos, K; Frost, F; Hunter, P R

    2015-10-01

    In this study the putative protective seroprevalence (PPS) of IgG antibodies to the 27-kDa and 15/17-kDa Cryptosporidium antigens in sera of healthy participants who were and were not exposed to Cryptosporidium oocysts via surface water-derived drinking water was compared. The participants completed a questionnaire regarding risk factors that have been shown to be associated with infection. The PPS was significantly greater (49-61%) in settlements where the drinking water originated from surface water, than in the control city where riverbank filtration was used (21% and 23%). Logistic regression analysis on the risk factors showed an association between bathing/swimming in outdoor pools and antibody responses to the 15/17-kDa antigen complex. Hence the elevated responses were most likely due to the use of contaminated water. Results indicate that waterborne Cryptosporidium infections occur more frequently than reported but may derive from multiple sources.

  2. Equine cryptosporidial infection associated with Cryptosporidium hedgehog genotype in Algeria.

    PubMed

    Laatamna, Abd Elkarim; Wagnerová, Pavla; Sak, Bohumil; Květoňová, Dana; Aissi, Miriem; Rost, Michael; Kváč, Martin

    2013-10-18

    Faecal samples from two horse farms in Algeria keeping Arabian, Thoroughbred, and Barb horses were examined for the presence of Cryptosporidium in 2010-2011. A total of 138 faecal samples (16 from a farm keeping 50 animals and 122 from a farm with 267 horses) were screened for Cryptosporidium spp. infection using molecular tools. DNA was extracted from all samples. Nested PCR was performed to amplify fragments of the SSU rDNA and gp60 genes to determine the presence of Cryptosporidium species and genotypes. Sequence analyses of SSU and gp60 genes revealed four animals positive for the presence of subtype XIIIa A22R9 of the Cryptosporidium hedgehog genotype. The infections were not associated with diarrhoea. This study reports, for the first time, the occurrence of Cryptosporidium in Algeria and the first occurrence of the hedgehog genotype in horses. These findings support the potential role of infected horses in sylvatic-domestic transmission of Cryptosporidium.

  3. Cryptosporidium species in Australian wildlife and domestic animals.

    PubMed

    Ryan, Una; Power, Michelle

    2012-11-01

    Cryptosporidium is an important enteric parasite that is transmitted via the fecal-oral route, water and food. Humans, wildlife and domestic livestock all potentially contribute Cryptosporidium to surface waters. Most species of Cryptosporidium are morphologically indistinguishable and can only be identified using molecular tools. Over 24 species have been identified and of these, 7 Cryptosporidium species/genotypes are responsible for most human cryptosporidiosis cases. In Australia, relatively few genotyping studies have been conducted. Six Cryptosporidium species (C. hominis, C. parvum, C. meleagridis, C. fayeri, C. andersoni and C. bovis) have been identified in humans in Australia. However, little is known about the contribution of animal hosts to human pathogenic strains of Cryptosporidium in drinking water catchments. In this review, we focus on the available genotyping data for native, feral and domestic animals inhabiting drinking water catchments in Australia to provide an improved understanding of the public health implications and to identify key research gaps.

  4. Open-source feature-tracking algorithm for sea ice drift retrieval from Sentinel-1 SAR imagery

    NASA Astrophysics Data System (ADS)

    Muckenhuber, Stefan; Andreevich Korosov, Anton; Sandven, Stein

    2016-04-01

    A computationally efficient, open-source feature-tracking algorithm, called ORB, is adopted and tuned for sea ice drift retrieval from Sentinel-1 SAR (Synthetic Aperture Radar) images. The most suitable setting and parameter values have been found using four Sentinel-1 image pairs representative of sea ice conditions between Greenland and Severnaya Zemlya during winter and spring. The performance of the algorithm is compared to two other feature-tracking algorithms, namely SIFT (Scale-Invariant Feature Transform) and SURF (Speeded-Up Robust Features). Having been applied to 43 test image pairs acquired over Fram Strait and the north-east of Greenland, the tuned ORB (Oriented FAST and Rotated BRIEF) algorithm produces the highest number of vectors (177 513, SIFT: 43 260 and SURF: 25 113), while being computationally most efficient (66 s, SIFT: 182 s and SURF: 99 s per image pair using a 2.7 GHz processor with 8 GB memory). For validation purposes, 314 manually drawn vectors have been compared with the closest calculated vectors, and the resulting root mean square error of ice drift is 563 m. All test image pairs show a significantly better performance of the HV (horizontal transmit, vertical receive) channel due to higher informativeness. On average, around four times as many vectors have been found using HV polarization. All software requirements necessary for applying the presented feature-tracking algorithm are open source to ensure a free and easy implementation.

  5. Tracking the MSL-SAM methane detection source location Through Mars Regional Atmospheric Modeling System (MRAMS)

    NASA Astrophysics Data System (ADS)

    Pla-García, Jorge

    2016-04-01

    olivine indicate could be the case, then it might explain the observed fast destruction of methane [7]. In an effort to better address the potential mixing and remaining questions, atmospheric circulation studies of Gale Crater were performed with the Mars Re-gional Atmospheric Modeling System (MRAMS). The model was focused on rover locations using nested grids with a spacing of 330 meters on the in-nermost grid that is centered over the landing [8, 9]. MRAMS is ideally suited for this investigation; the model is explicitly designed to simulate Mars' at-mospheric circulations at the mesoscale and smaller with realistic, high-resolution surface properties [10, 11]. In order to characterize seasonal mixing changes throughout the Martian year, simulations were con-ducted at Ls 0, 90, 180 and 270. Two additional sim-ulations at Ls 225 and 315 were explored to better understand the unique meteorological setting cen-tered around Ls 270. Ls 270 was shown to be an anomalous season when air within and outside the crater was well mixed by strong, flushing, northerly flow and large amplitude breaking mountain waves: air flowing downslope at night is cold enough to penetrate all the way to the surface. At other seasons, the air in the crater is more isolated -but not com-pletely- from the surrounding environment: mesoscale simulations indicate that the air flowing down the crater rims does not easily make it to the crater floor. Instead, the air encounters very cold and stable air pooled in the bottom of the crater, which forces the air to glide right over the colder, more dense air below. Thus, the mixing of near-surface crater air with the external environment is potentially more limited than around Ls 270. 2. Tracking methane source location The rise in concentration was reported to start around sol 300 (˜Ls 336), peaked shortly after sol 520 (˜Ls 82), and then dropped to background val-ues prior to sol 575 (˜Ls 103). Two scenarios are considered in the context of the

  6. Die-off rates of Cryptosporidium parvum oocysts in a swine lagoon and in a spray field

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Because of several large-scale outbreaks of cryptosporidiosis in humans, Cryptosporidium has become a public health concern. Commercial swine operations apply large volumes of effluent from lagoons to spray fields as a waste management practice. This effluent is a source of Cryptosporidi...

  7. Quantification of hsp70 mRNA from the Cryptosporidium parvum in soil by reverse transcription real-time PCR

    EPA Science Inventory

    As one of the leading causes of waterborne enteric disease, Cryptosporidium parvum poses significant threat to public health. Besides water, soil can also become an important environmental source of C. parvum once polluted. Detection of viable C. parvum in soil is a key issue whe...

  8. Characterization of sources and loadings of fecal pollutants using microbial source tracking assays in urban and rural areas of the Grand River Watershed, Southwestern Ontario.

    PubMed

    Lee, Dae-Young; Lee, Hung; Trevors, Jack T; Weir, Susan C; Thomas, Janis L; Habash, Marc

    2014-04-15

    Sources of fecal water pollution were assessed in the Grand River and two of its tributaries (Ontario, Canada) using total and host-specific (human and bovine) Bacteroidales genetic markers in conjunction with reference information, such as land use and weather. In-stream levels of the markers and culturable Escherichia coli were also monitored during multiple rain events to gain information on fecal loadings to catchment from diffuse sources. Elevated human-specific marker levels were accurately identified in river water impacted by a municipal wastewater treatment plant (WWTP) effluent and at a downstream site in the Grand River. In contrast, the bovine-specific marker showed high levels of cattle fecal pollution in two tributaries, both of which are characterized as intensely farmed areas. The bovine-specific Bacteroidales marker increased with rainfall in the agricultural tributaries, indicating enhanced loading of cattle-derived fecal pollutants to river from non-point sources following rain events. However, rain-triggered fecal loading was not substantiated in urban settings, indicating continuous inputs of human-originated fecal pollutants from point sources, such as WWTP effluent. This study demonstrated that the Bacteroidales source tracking assays, in combination with land use information and hydrological data, may provide additional insight into the spatial and temporal distribution of source-specific fecal contamination in streams impacted by varying land uses. Using the approach described in this study may help to characterize impacted water sources and to design targeted land use management plans in other watersheds in the future.

  9. Sources of variation in assessing left atrial functions by 2D speckle-tracking echocardiography.

    PubMed

    Rimbaş, Roxana Cristina; Mihăilă, Sorina; Vinereanu, Dragoş

    2016-03-01

    Left atrial (LA) strain and strain rate, determined by speckle-tracking echocardiography (STE), are reproducible indices to assess LA function. Different normal ranges for LA phasic functions have been reported. We investigated the role of the reference point (P- and R-wave), gain, and region of interest (ROI), as the major sources of variation when assessing LA function. 52 subjects were evaluated for LA conventional and STE analysis. 45 of them (46 ± 14 years, 26 men) were feasible for concomitant LA deformation, and LA phasic volumes and ejection fractions (LAEF) evaluation. First, we compared the P- and R-wave methods, for the evaluation of the LA functions. We used diastolic mitral profile to clearly delineate the time intervals for each LA function. For the P-wave method, active function was assessed from negative global strain as a difference between the strain at pre-atrial contraction and strain just before mitral valve closure (GSA-), and late diastolic strain rate (GSRL); passive function from positive strain at MVO (GSA+), and from early negative diastolic strain rate (GSRE); reservoir function from the sum of GSA- and GSA+ (TGSA), and positive strain rate at the beginning of LV systole (GSR+). For the R-wave method we used the same SR parameters. The active function was evaluated by late positive global strain (GSAC), the reservoir by positive peak before the opening of the mitral valve (TGSA), and conduit function by the difference between TGSA and GSAC (GSA+). Then, by using P-wave method, we measured all previously described parameters for different gains-minimum (G0), medium (G12), and maximum (G24), and for different ROIs-minimum (ROI0), step 1 (ROI1), and 2 (ROI2). Feasibility of the LA strain measurements was 87 %. Active LA function was similar in the absolute value (GSAC and GSA-), whereas passive and reservoir functions were significantly higher (GSA+, TGSA) with the R-wave method. Active LAEF correlated with GSA- measured by the P-wave (r

  10. Concentrations, viability, and distribution of Cryptosporidium genotypes in lagoons of swine facilities in the Southern Piedmont and in coastal plain watersheds of Georgia.

    PubMed

    Jenkins, Michael B; Liotta, Janice L; Lucio-Forster, Araceli; Bowman, Dwight D

    2010-09-01

    Waste lagoons of swine operations are a source of Cryptosporidium oocysts. Few studies, however, have reported on oocyst concentrations in swine waste lagoons; none have reported on oocyst viability status, nor has there been a systematic assessment of species/genotype distributions across different types of swine facilities. Ten swine waste lagoons associated with farrowing, nursery, finishing, and gestation operations were each sampled once a month for a year. Oocysts were extracted from triplicate 900-ml effluent samples, enumerated by microscopy, and assessed for viability by dye exclusion/vital stain assay. DNA was extracted from processed samples, and 18S ribosomal DNA (rDNA) genes were amplified by PCR and sequenced for species and genotype identification. Oocysts were observed at each sampling time at each lagoon. Annual mean concentrations of total oocysts and viable oocysts ranged between 24 and 51 and between 0.6 and 12 oocysts ml(-1) effluent, respectively. The species and genotype distributions were dominated (95 to 100%) by Cryptosporidium suis and Cryptosporidium pig genotype II, the latter of which was found at eight of the lagoons. The lagoon at the gestation facility was dominated by Cryptosporidium muris (90%), and one farrowing facility showed a mix of pig genotypes, Cryptosporidium muris, and various genotypes of C. parvum. The zoonotic C. parvum bovine genotype was observed five times out of 407 18S rDNA sequences analyzed. Our results indicate that pigs can have mixed Cryptosporidium infections, but infection with C. suis is likely to be dominant.

  11. Prevalence of Cryptosporidium species and genotypes in mature dairy cattle on farms in eastern United States compared with younger cattle from the same locations.

    PubMed

    Fayer, Ronald; Santin, Monica; Trout, James M

    2007-04-30

    Feces collected from 541 milking cows on two dairy farms each in Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida were examined for the presence of Cryptosporidium oocysts. Oocysts were concentrated from 15 g of feces from each cow and DNA was extracted. A two-step nested PCR protocol was used to amplify an 830 base pair fragment of the SSUrRNA gene. PCR-positive products were purified and sequenced. PCR-positive findings were obtained from cows in all seven states and from 11 of 14 farms. Cryptosporidium parvum, Cryptosporidium bovis, and Cryptosporidium andersoni were found on 2, 6, and 8 farms, and infected 0.4, 1.7, and 3.7% of the 541 cows, respectively. The overall lower prevalence of Cryptosporidium in these cows was very highly significant (p< or =0.0001) compared with younger cattle and the relative prevalence of each species of Cryptosporidium also differed when compared with younger cattle previously examined on most of these same farms. The very low level of infection with C. parvum, the major species pathogenic to both cattle and humans, suggests that mature dairy cattle are a relatively low risk source of infection for humans.

  12. Fecal pollution source tracking in waters intended for human supply based on archaeal and bacterial genetic markers.

    PubMed

    Bianco, Kayo; Barreto, Camila; Oliveira, Samara Sant'Anna; Pinto, Leonardo Henriques; Albano, Rodolpho Mattos; Miranda, Catia Chaia; Clementino, Maysa Mandetta

    2015-12-01

    The determination of fecal pollution sources in aquatic ecosystems is essential to estimate associated health risks. In this study, we evaluate eight microbial source tracking (MST) markers including host-specific Bacteroidales and Methanobrevibacter spp. for discrimination between human, bovine, equine, and swine fecal contamination in waters intended for human supply. Overall, the novel host-specific archaeal and bacterial primers proposed in this study demonstrated high sensitivity and specificity. Markers for the Archaea domain were more prevalent in the fecal and water samples studied. We conclude that the investigations regarding the sources of fecal pollution in public water supplies can contribute to improve the quality of human health. To our knowledge, this is the first analysis using both archaeal and bacterial fecal MST markers on tropical water bodies of Rio de Janeiro city, Brazil.

  13. Stardust Interstellar Preliminary Examination II: Curating the Interstellar Dust Collector, Picokeystones, and Sources of Impact Tracks

    NASA Technical Reports Server (NTRS)

    Frank, David R.; Westphal, Andrew J.; Zolensky, Michael E.; Gainsforth, Zack; Butterworth, Anna L.; Bastien, Ronald K.; Allen, Carlton; Anderson, David; Bechtel, Hans A.; Sandford, Scott A.

    2013-01-01

    We discuss the inherent difficulties that arise during "ground truth" characterization of the Stardust interstellar dust collector. The challenge of identifying contemporary interstellar dust impact tracks in aerogel is described within the context of background spacecraft secondaries and possible interplanetary dust particles and beta-meteoroids. In addition, the extraction of microscopic dust embedded in aerogel is technically challenging. Specifically, we provide a detailed description of the sample preparation techniques developed to address the unique goals and restrictions of the Interstellar Preliminary Exam. These sample preparation requirements and the scarcity of candidate interstellar impact tracks exacerbate the difficulties. We also illustrate the role of initial optical imaging with critically important examples, and summarize the overall processing of the collection to date.

  14. Cryptosporidium spp. and Giardia spp. in surface water supply of Campinas, southeast Brazil.

    PubMed

    Neto, Romeu Cantusio; dos Santos, Luciana Urbano; Sato, Maria Ines Zanoli; Franco, Regina Maura Bueno

    2010-01-01

    Surface water contaminated by domestic sewage discharges is a potential source of pathogens, including protozoa. During 2005-2006, the source water (Atibaia River) of the Surface Water Treatment Plant (WTP) of Campinas city, São Paulo, Brazil was sampled to obtain an assessment of Cryptosporidium oocyst and Giardia cyst concentrations. Calcium carbonate flocculation (CCF) and membrane filtration (MF) concentration techniques, with and without purification by immunomagnetic separation (IMS) were evaluated. The cysts and oocysts were detected by immunofluorescence assay (IFA) and confirmed by differential interference contrast (DIC). Membrane filtration method generally produced higher recovery efficiency. Giardia spp. was detected in 87.5% of the water samples analyzed with densities ranging from 2.5 to 120 cysts per L. Cryptosporidium spp were detected in 62.5% and the concentrations ranged from 15 to 60 oocysts per L. Cryptosporidium oocyst and Giardia cyst concentrations detected in this study were elevated and are associated with discharge of untreated sewage in Atibaia River. Measures should be taken to protect surface water from sources of contamination.

  15. Comparison of Microbial and Chemical Source Tracking Markers To Identify Fecal Contamination Sources in the Humber River (Toronto, Ontario, Canada) and Associated Storm Water Outfalls.

    PubMed

    Staley, Zachery R; Grabuski, Josey; Sverko, Ed; Edge, Thomas A

    2016-11-01

    Storm water runoff is a major source of pollution, and understanding the components of storm water discharge is essential to remediation efforts and proper assessment of risks to human and ecosystem health. In this study, culturable Escherichia coli and ampicillin-resistant E. coli levels were quantified and microbial source tracking (MST) markers (including markers for general Bacteroidales spp., human, ruminant/cow, gull, and dog) were detected in storm water outfalls and sites along the Humber River in Toronto, Ontario, Canada, and enumerated via endpoint PCR and quantitative PCR (qPCR). Additionally, chemical source tracking (CST) markers specific for human wastewater (caffeine, carbamazepine, codeine, cotinine, acetaminophen, and acesulfame) were quantified. Human and gull fecal sources were detected at all sites, although concentrations of the human fecal marker were higher, particularly in outfalls (mean outfall concentrations of 4.22 log10 copies, expressed as copy numbers [CN]/100 milliliters for human and 0.46 log10 CN/100 milliliters for gull). Higher concentrations of caffeine, acetaminophen, acesulfame, E. coli, and the human fecal marker were indicative of greater raw sewage contamination at several sites (maximum concentrations of 34,800 ng/liter, 5,120 ng/liter, 9,720 ng/liter, 5.26 log10 CFU/100 ml, and 7.65 log10 CN/100 ml, respectively). These results indicate pervasive sewage contamination at storm water outfalls and throughout the Humber River, with multiple lines of evidence identifying Black Creek and two storm water outfalls with prominent sewage cross-connection problems requiring remediation. Limited data are available on specific sources of pollution in storm water, though our results indicate the value of using both MST and CST methodologies to more reliably assess sewage contamination in impacted watersheds.

  16. High sensitive gas detection and isotopic measurement for the applications of industrial emission online monitoring and air pollution source tracking

    NASA Astrophysics Data System (ADS)

    Dong, Fengzhong; Zhang, Zhirong; Xia, Hua; Cui, Xiaojuan; Pang, Tao; Wu, Bian; Chen, Weidong; Sigrist, Markus

    2015-04-01

    High sensitive gas detection and isotopic measurements have been widely employed in the industrial and safety production. The recent progress made by our group on high sensitive gas detection with technologies of TDLAS, off-axis integrated cavity output spectroscopy (OA-ICOS) and cavity ring-down spectroscopy (CRDS) will be briefly summarized in this report. Some works for field applications of industrial emission online monitoring and gas leakage detection in oil tank farm with TDLAS are first presented, and then part of our most recent researches on isotopic gas detection with OA-ICOS and CRDS for tracking of pollution sources are also introduced.

  17. The Applicability of TaqMan-Based Quantitative Real-Time PCR Assays for Detecting and Enumerating Cryptosporidium spp. Oocysts in the Environment

    PubMed Central

    Staggs, Sarah E.; Beckman, Erin M.; Keely, Scott P.; Mackwan, Reena; Ware, Michael W.; Moyer, Alan P.; Ferretti, James A.; Sayed, Abu; Xiao, Lihua; Villegas, Eric N.

    2013-01-01

    Quantitative real-time polymerase chain reaction (qPCR) assays to detect Cryptosporidium oocysts in clinical samples are increasingly being used to diagnose human cryptosporidiosis, but a parallel approach for detecting and identifying Cryptosporidium oocyst contamination in surface water sources has yet to be established for current drinking water quality monitoring practices. It has been proposed that Cryptosporidium qPCR-based assays could be used as viable alternatives to current microscopic-based detection methods to quantify levels of oocysts in drinking water sources; however, data on specificity, analytical sensitivity, and the ability to accurately quantify low levels of oocysts are limited. The purpose of this study was to provide a comprehensive evaluation of TaqMan-based qPCR assays, which were developed for either clinical or environmental investigations, for detecting Cryptosporidium oocyst contamination in water. Ten different qPCR assays, six previously published and four developed in this study were analyzed for specificity and analytical sensitivity. Specificity varied between all ten assays, and in one particular assay, which targeted the Cryptosporidium 18S rRNA gene, successfully detected all Cryptosporidium spp. tested, but also cross-amplified T. gondii, fungi, algae, and dinoflagellates. When evaluating the analytical sensitivity of these qPCR assays, results showed that eight of the assays could reliably detect ten flow-sorted oocysts in reagent water or environmental matrix. This study revealed that while a qPCR-based detection assay can be useful for detecting and differentiating different Cryptosporidium species in environmental samples, it cannot accurately measure low levels of oocysts that are typically found in drinking water sources. PMID:23805235

  18. ICR SS protozoan data site-by-site: a picture of Cryptosporidium and Giardia in U.S. surface water.

    PubMed

    Ongerth, Jerry E

    2013-09-17

    The U.S. Environmental Protection Agency (USEPA) Information Collection Rule Supplemental Survey (ICR SS) required analysis of Cryptosporidium and Giardia in 10 L surface water samples twice a week for a year by USEPA Method 1623 at 80 representative U.S. public water systems (PWS). The resulting data are examined site-by-site in relation to objectives of the Federal drinking water regulation, The Long-Term (2) Enhanced Surface Water Treatment Rule (LT2), currently under formal 6-year review by the USEPA. The data describe Cryptosporidium and Giardia in watersheds nation-wide over a single annual cycle. Due to limited recovery efficiency measurement results are not fully quantified. In the required sample volumes of 10 L no Cryptosporidium were found in 86% of samples and no Giardia were found in 67% of samples. Yet, organisms were found in enough samples at 34 of 80 sites to detail a specrtum of occurrence and variability for both organisms. The data are shown to describe indivudual site risk essential for guidance of watershed and water treatment management by PWSs. The span of median occurrence for both organisms was about 2 orders of magnitude above the limit of detection (LD), ca. 0.05 raw no's/L for Cryptosporidium and ca. 0.10 raw no's/L for Giardia. Data analysis illustrates key features of Cryptosporidium and Giardia in surface water: presence is continuous not intermittent; zeros indicate presence below the LD; occurrence level and variations depend on watershed sources; risk depends on both magnitude and variability of concentration; accurate estimation of risk requires routine measurement of recovery efficiency and calculation of concentration. The data and analysis illustrate features of Cryptosporidium and Giardia occurrence in surface water relevant to their effective regulation for public health protection.

  19. Human and Animal Fecal Contamination of Community Water Sources, Stored Drinking Water and Hands in Rural India Measured with Validated Microbial Source Tracking Assays

    PubMed Central

    Schriewer, Alexander; Odagiri, Mitsunori; Wuertz, Stefan; Misra, Pravas R.; Panigrahi, Pinaki; Clasen, Thomas; Jenkins, Marion W.

    2015-01-01

    We examined pathways of exposure to fecal contamination of human and animal origin in 24 villages in Odisha, India. In a cross-sectional study during the monsoon season, fecal exposure via community water sources (N = 123) and in the home (N = 137) was assessed using human- and nonhuman-associated Bacteroidales microbial source tracking (MST) markers and fecal coliforms (FCs). Detection rates and marker concentrations were examined to pinpoint pathways of human fecal exposure in the public and domestic domains of disease transmission in study communities. Human fecal markers were detected much more frequently in the domestic domain (45% of households) than in public domain sources (8% of ponds; 4% of groundwater drinking sources). Animal fecal markers were widely detected in both domains (74% of ponds, 96% of households, 10% of groundwater drinking sources), indicating ubiquitous risks of exposure to animal feces and zoonotic pathogens. This study confirms an often suggested contamination link from hands to stored water in the home in developing countries separately for mothers' and children's hands and both human and animal fecal contamination. In contrast to MST markers, FCs provided a poor metric to assess risks of exposure to fecal contamination of human origin in this rural setting. PMID:26149868

  20. Human and Animal Fecal Contamination of Community Water Sources, Stored Drinking Water and Hands in Rural India Measured with Validated Microbial Source Tracking Assays.

    PubMed

    Schriewer, Alexander; Odagiri, Mitsunori; Wuertz, Stefan; Misra, Pravas R; Panigrahi, Pinaki; Clasen, Thomas; Jenkins, Marion W

    2015-09-01

    We examined pathways of exposure to fecal contamination of human and animal origin in 24 villages in Odisha, India. In a cross-sectional study during the monsoon season, fecal exposure via community water sources (N = 123) and in the home (N = 137) was assessed using human- and nonhuman-associated Bacteroidales microbial source tracking (MST) markers and fecal coliforms (FCs). Detection rates and marker concentrations were examined to pinpoint pathways of human fecal exposure in the public and domestic domains of disease transmission in study communities. Human fecal markers were detected much more frequently in the domestic domain (45% of households) than in public domain sources (8% of ponds; 4% of groundwater drinking sources). Animal fecal markers were widely detected in both domains (74% of ponds, 96% of households, 10% of groundwater drinking sources), indicating ubiquitous risks of exposure to animal feces and zoonotic pathogens. This study confirms an often suggested contamination link from hands to stored water in the home in developing countries separately for mothers' and children's hands and both human and animal fecal contamination. In contrast to MST markers, FCs provided a poor metric to assess risks of exposure to fecal contamination of human origin in this rural setting.

  1. Acute enterocolitis in a human being infected with the protozoan Cryptosporidium.

    PubMed

    Nime, F A; Burek, J D; Page, D L; Holscher, M A; Yardley, J H

    1976-04-01

    A 3-year-old child with severe acute self-limited enterocolitis was found on rectal biopsy to be infected with the protozoal parasite Cryptosporidium. This organism is known to infect a variety of vertebrates, but this is the first report of infection by Cryptosporidium in a human being. Both light and electron microscopic findings in the rectal biopsy are reported. It is suggested, on the basis of the severity of the clinical symptoms, and on the pathological changes in the rectum, that the organism in this case is likely to have been the cause of the enterocolitis and thus to have been a pathogen rather than a commensal. The source of the infection in this child could not be established. The value of signoidoscopy and biopsies is noted in this condition and as a general method for determining the etiology of a gastrointestinal infection in cases where other studies are negative.

  2. EVALUATING CRYPTOSPORIDIUM AND GIARDIA CONCENTRATIONS IN COMBINED SEWER OVERFLOW

    EPA Science Inventory

    Since the first identified Cryptosporidium outbreaks occurred in the 1980s and the massive 1993 Milwaukee, WI outbreak affected more than 400,000 people (Fox & Lytle 1996), the concern over the public health risks linked to protozoan pathogens Cryptosporidium and Giardia has grow...

  3. Tracking sources of Staphylococcus aureus hand contamination in food handlers by spa typing.

    PubMed

    Ho, Jeffery; Boost, Maureen V; O'Donoghue, Margaret M

    2015-07-01

    We aimed to identify the source of Staphylococcus aureus contaminating hands of food handlers. Nasal samples and direct fingertip imprints were collected on 2 occasions from food handlers and characterized to determine likely sources of hand contamination. Most hand contamination was attributable to nasal isolates of persistently colonized coworkers who had presumably contaminated the environment. Regular handwashing should be supplemented by effective environmental disinfection.

  4. DEVELOPMENT OF HOST-SPECIFIC METAGENOMIC MARKERS FOR MICROBIAL SOURCE TRACKING USING A NOVEL METAGENOMIC APPROACH

    EPA Science Inventory

    Fecal contamination of source waters is an important issue to the drinking water industry. Improper disposal of animal waste, leaky septic tanks, storm runoff, and wildlife can all be responsible for spreading enteric pathogens into source waters. As a result, methods that can pi...

  5. FECAL BACTERIA SOURCE TRACKING AND BACTEROIDES SPP. HOST SPECIES SPECIFICITY ANALYSIS

    EPA Science Inventory

    Point and non-point pollution sources of fecal pollution on a watershed adversely impact the quality of drinking source waters and recreational waters. States are required to develop total maximum daily loads (TMDLs) and devise best management practices (BMPs) to reduce the po...

  6. Application of iron and zinc isotopes to track the sources and mechanisms of metal loading in a mountain watershed

    USGS Publications Warehouse

    Borrok, D.M.; Wanty, R.B.; Ian, Ridley W.; Lamothe, P.J.; Kimball, B.A.; Verplanck, P.L.; Runkel, R.L.

    2009-01-01

    Here the hydrogeochemical constraints of a tracer dilution study are combined with Fe and Zn isotopic measurements to pinpoint metal loading sources and attenuation mechanisms in an alpine watershed impacted by acid mine drainage. In the tested mountain catchment, ??56Fe and ??66Zn isotopic signatures of filtered stream water samples varied by ???3.5??? and 0.4???, respectively. The inherent differences in the aqueous geochemistry of Fe and Zn provided complimentary isotopic information. For example, variations in ??56Fe were linked to redox and precipitation reactions occurring in the stream, while changes in ??66Zn were indicative of conservative mixing of different Zn sources. Fen environments contributed distinctively light dissolved Fe (<-2.0???) and isotopically heavy suspended Fe precipitates to the watershed, while Zn from the fen was isotopically heavy (>+0.4???). Acidic drainage from mine wastes contributed heavier dissolved Fe (???+0.5???) and lighter Zn (???+0.2???) isotopes relative to the fen. Upwelling of Fe-rich groundwater near the mouth of the catchment was the major source of Fe (??56Fe ??? 0???) leaving the watershed in surface flow, while runoff from mining wastes was the major source of Zn. The results suggest that given a strong framework for interpretation, Fe and Zn isotopes are useful tools for identifying and tracking metal sources and attenuation mechanisms in mountain watersheds. ?? 2009 Elsevier Ltd.

  7. Occurrence and potential health risk of Cryptosporidium and Giardia in the Three Gorges Reservoir, China.

    PubMed

    Xiao, Guosheng; Qiu, Zhiqun; Qi, Junsheng; Chen, Ji-an; Liu, Fengdan; Liu, Wenyi; Luo, Jiaohua; Shu, Weiqun

    2013-05-01

    The Three Gorges Reservoir (TGR) is the biggest lake in the world and a major water source in China. There is no information about occurrence and impact of Cryptosporidium and Giardia on the aquatic ecosystem. 61 surface water samples from 23 monitoring sites and 5 treated effluent samples were collected and analyzed. Cryptosporidium oocysts and Giardia cysts were found, respectively, in 86.4% and 65.2% of a total of 66 water samples, with high concentrations in treated effluent. The mean percent recovery was 29.14% for oocysts and 34.86% for cysts. A seasonal pattern was observed, with positive samples for Cryptosporidium more frequent in flood period and positive samples for Giardia more frequent in impounding period. Counts of enterococci, fecal coliforms and total coliforms, and turbidity were significantly associated with Cryptosporidium concentration in backwater (water in a main river which is backed up by the Three Gorges Dam) areas of tributaries but not Giardia. High associations were also found between oocyst and cyst in backwater areas of tributaries and cities. The risks of infection and illness due to water consumption in four different exposure routes were estimated. The results showed that swimming in the TGR has the highest infection risk with 1.39 × 10(-3) per time (95% confidence interval (CI): 0.05-600.3 × 10(-5)) for Cryptosporidium and 2.08 × 10(-4) per time (95% CI: 0.05-878.87 × 10(-6)) for Giardia, while directly drinking unboiled tap water treated with the conventional process has the highest morbidity with 524.98 per 100,000 population per year (95% CI: 10.35-2040.26) for Cryptosporidium and 5.89 per 100,000 population per year (95% CI: 0.08-22.67) for Giardia. This study provides new useful information for drinking water plants, health care workers and managers to improve the safety of tap water and deduce the risk of surface water contamination in China.

  8. Quantitative risk assessment of Cryptosporidium in tap water in Ireland.

    PubMed

    Cummins, E; Kennedy, R; Cormican, M

    2010-01-15

    Cryptosporidium species are protozoan parasites associated with gastro-intestinal illness. Following a number of high profile outbreaks worldwide, it has emerged as a parasite of major public health concern. A quantitative Monte Carlo simulation model was developed to evaluate the annual risk of infection from Cryptosporidium in tap water in Ireland. The assessment considers the potential initial contamination levels in raw water, oocyst removal and decontamination events following various process stages, including coagulation/flocculation, sedimentation, filtration and disinfection. A number of scenarios were analysed to represent potential risks from public water supplies, group water schemes and private wells. Where surface water is used additional physical and chemical water treatment is important in terms of reducing the risk to consumers. The simulated annual risk of illness for immunocompetent individuals was below 1 x 10(-4) per year (as set by the US EPA) except under extreme contamination events. The risk for immunocompromised individuals was 2-3 orders of magnitude greater for the scenarios analysed. The model indicates a reduced risk of infection from tap water that has undergone microfiltration, as this treatment is more robust in the event of high contamination loads. The sensitivity analysis highlighted the importance of watershed protection and the importance of adequate coagulation/flocculation in conventional treatment. The frequency of failure of the treatment process is the most important parameter influencing human risk in conventional treatment. The model developed in this study may be useful for local authorities, government agencies and other stakeholders to evaluate the likely risk of infection given some basic input data on source water and treatment processes used.

  9. Surveillance Systems for Waterborne Protozoa Past, Present and Future

    EPA Science Inventory

    OVERVIEW I. Brief introduction to waterborne Cryptosporidium Historical perspective on detecting Cryptosporidium Current detection methodologies II. US EPA’s waterborne protozoan research program Detecting, typing, and tracking sources of Cryptosporidium contami...

  10. Incorporating expert judgments in utility evaluation of bacteroidales qPCR assays for microbial source tracking in a drinking water source.

    PubMed

    Åström, Johan; Pettersson, Thomas J R; Reischer, Georg H; Norberg, Tommy; Hermansson, Malte

    2015-02-03

    Several assays for the detection of host-specific genetic markers of the order Bacteroidales have been developed and used for microbial source tracking (MST) in environmental waters. It is recognized that the source-sensitivity and source-specificity are unknown and variable when introducing these assays in new geographic regions, which reduces their reliability and use. A Bayesian approach was developed to incorporate expert judgments with regional assay sensitivity and specificity assessments in a utility evaluation of a human and a ruminant-specific qPCR assay for MST in a drinking water source. Water samples from Lake Rådasjön were analyzed for E. coli, intestinal enterococci and somatic coliphages through cultivation and for human (BacH) and ruminant-specific (BacR) markers through qPCR assays. Expert judgments were collected regarding the probability of human and ruminant fecal contamination based on fecal indicator organism data and subjective information. Using Bayes formula, the conditional probability of a true human or ruminant fecal contamination given the presence of BacH or BacR was determined stochastically from expert judgments and regional qPCR assay performance, using Beta distributions to represent uncertainties. A web-based computational tool was developed for the procedure, which provides a measure of confidence to findings of host-specific markers and demonstrates the information value from these assays.

  11. Cryptosporidium galli and novel Cryptosporidium avian genotype VI in North American red-winged blackbirds (Agelaius phoeniceus).

    PubMed

    Chelladurai, Jeba Jesudoss; Clark, Mark E; Kváč, Martin; Holubová, Nikola; Khan, Eakalak; Stenger, Brianna L S; Giddings, Catherine W; McEvoy, John

    2016-05-01

    Proventriculus and intestinal samples from 70 North American red-winged blackbirds (Agelaius phoeniceus; order Passeriformes) were examined for the presence of Cryptosporidium by PCR amplification and sequence analysis of the 18S ribosomal RNA (18S rRNA), actin, and 70-kDa heat shock protein (HSP70) genes. Twelve birds (17.1 %) were positive for the Cryptosporidium 18S rRNA gene: six birds were positive at the proventriculus site only and six birds were positive at the proventriculus and intestinal sites. Sequence analysis of the 18S rRNA, actin and HSP70 genes showed the presence of the gastric species Cryptosporidium galli in a single proventriculus sample and a closely related genotype, which we have named Cryptosporidium avian genotype VI, in all other positive samples. These findings contribute to our understanding of Cryptosporidium diversification in passerines, the largest avian order.

  12. Microbial source tracking markers for detection of fecal contamination in environmental waters: relationships between pathogens and human health outcomes.

    PubMed

    Harwood, Valerie J; Staley, Christopher; Badgley, Brian D; Borges, Kim; Korajkic, Asja

    2014-01-01

    Microbial source tracking (MST) describes a suite of methods and an investigative strategy for determination of fecal pollution sources in environmental waters that rely on the association of certain fecal microorganisms with a particular host. MST is used to assess recreational water quality and associated human health risk, and total maximum daily load allocations. Many methods rely on signature molecules (markers) such as DNA sequences of host-associated microorganisms. Human sewage pollution is among the greatest concerns for human health due to (1) the known risk of exposure to human waste and (2) the public and regulatory will to reduce sewage pollution; however, methods to identify animal sources are receiving increasing attention as our understanding of zoonotic disease potential improves. Here, we review the performance of MST methods in initial reports and field studies, with particular emphasis on quantitative PCR (qPCR). Relationships among human-associated MST markers, fecal indicator bacteria, pathogens, and human health outcomes are presented along with recommendations for future research. An integrated understanding of the advantages and drawbacks of the many MST methods targeting human sources advanced over the past several decades will benefit managers, regulators, researchers, and other users of this rapidly growing area of environmental microbiology.

  13. Signal information available for plume source tracking with and without surface waves and learning by undergraduates assisting with the research

    NASA Astrophysics Data System (ADS)

    Wiley, Megan Beth

    Autonomous vehicles have had limited success in locating point sources of pollutants, chemicals, and other passive scalars. However, animals such as stomatopods, a mantis shrimp, track odor plumes easily for food, mates, and habitat. Laboratory experiments using Planar Laser Induced Fluorescence measured odor concentration downstream of a diffusive source with and without live stomatopods to investigate their source-tracking strategies in unidirectional and "wave-affected" (surface waves with a mean current) flows. Despite the dearth of signal, extreme temporal variation, and meandering plume centerline, the stomatopods were able to locate the source, especially in the wave-affected flow. Differences in the two plumes far from the source (>160 cm) appeared to help the animals in the wave-affected flow position themselves closer to the source (<70 cm) at times with relatively large amounts of odor and plume filaments of high concentration. At the height of the animals' antennules, the site of their primary chemosensors, the time-averaged Reynolds stresses in the two flows were approximately the same. The temporal variation in stresses over the wave cycle may be responsible for differences in the two plumes. The antennule height falls between a region of large peaks in Reynolds stress in phase with peaks in streamwise acceleration, and a lower region with a smaller Reynolds stress peak in phase with maximum shear during flow reversal. Six undergraduate students assisted with the research. We documented their daily activities and ideas on plume dispersion using open-ended interviews. Most of their time was spent on tasks that required no understanding of fluid mechanics, and there was little evidence of learning by participation in the RAship. One RA's conceptions of turbulence did change, but a group workshop seemed to support this learning more than the RAship. The documented conceptions could aid in curriculum design, since situating new information within current

  14. Biomarkers in sedimentary sequences: Indicators to track sediment sources over decadal timescales

    NASA Astrophysics Data System (ADS)

    Chen, F. X.; Fang, N. F.; Wang, Y. X.; Tong, L. S.; Shi, Z. H.

    2017-02-01

    Long-term sedimentary sequence research can reveal how human activities and climate interact to affect catchment vegetation, flooding, soil erosion, and sediment sources. In this study, a biomarker sediment fingerprinting technique based on n-alkanes was used to identify long timescale (decadal) sediment sources in a small agricultural catchment. However, the highly saline carbonate environment and bacterial and algal activities elevated the levels of even-chain n-alkanes in the sediments, leading to an obvious even-over-odd predominance of short and middle components (C15-C26). Therefore, by analyzing three odd, long-chain n-alkanes (C27, C29 and C31) in 27 source samples from cropland, gully, and steep slope areas and one sediment sequence (one cultivated horizon and 47 flood couplets), a composite fingerprinting method and genetic algorithm optimization were applied to find the optimal source contributions to sediments. The biomarker fingerprinting results demonstrated that the primary sediment source is gullies, followed by cropland and steep slope areas. The average median source contributions associated with 47 flood couples collected from sediment core samples ranged from 0 ± 0.1% to 91.9 ± 0.4% with an average of 45.0% for gullies, 0 ± 0.4% to 95.6 ± 1.6% with an average of 38.2% for cropland, and 0 ± 2.1% to 60.7 ± 0.4% with an average of 16.8% for steep slopes. However, because farmers were highly motivated to manage the cropland after the 1980s, over half the sediments were derived from cropland in the 1980s. Biomarkers have significant advantages in the identification of sediments derived from different landscape units (e.g., gully and steep slope areas), and n-alkanes have considerable potential in high-resolution research of environmental change based on soil erosion in the hilly Loess Plateau region.

  15. Multicentric Evaluation of a New Real-Time PCR Assay for Quantification of Cryptosporidium spp. and Identification of Cryptosporidium parvum and Cryptosporidium hominis

    PubMed Central

    Chapey, E.; Dutoit, E.; Guyot, K.; Hasseine, L.; Jeddi, F.; Menotti, J.; Paraud, C.; Pomares, C.; Rabodonirina, M.; Rieux, A.; Derouin, F.

    2013-01-01

    Cryptosporidium is a protozoan parasite responsible for gastroenteritis, especially in immunocompromised patients. Laboratory diagnosis of cryptosporidiosis relies on microscopy, antigen detection, and nucleic acid detection and analysis. Among the numerous molecular targets available, the 18S rRNA gene displays the best sensitivity and sequence variations between species and can be used for molecular typing assays. This paper presents a new real-time PCR assay for the detection and quantification of all Cryptosporidium species associated with the identification of Cryptosporidium hominis and Cryptosporidium parvum. The sensitivity and specificity of this new PCR assay were assessed on a multicentric basis, using well-characterized Cryptosporidium-positive and -negative human stool samples, and the efficiencies of nine extraction methods were comparatively assessed using Cryptosporidium-seeded stool samples and phosphate-buffered saline samples. A comparison of extraction yields showed that the most efficient extraction method was the Boom technique in association with mechanical grinding, and column extraction showed higher binding capacity than extraction methods based on magnetic silica. Our PCR assay was able to quantify at least 300 oocysts per gram of stool. Satisfactory reproducibility between laboratories was observed. The two main species causing human disease, Cryptosporidium hominis and Cryptosporidium parvum, were identified using a duplex real-time PCR assay with specific TaqMan minor-groove-binding ligand (MGB) probes for the same amplicon. To conclude, this one-step quantitative PCR is well suited to the routine diagnosis of cryptosporidiosis since practical conditions, including DNA extraction, quantification using well-defined standards, and identification of the two main species infecting humans, have been positively assessed. PMID:23720792

  16. Effectiveness of water treatment for the removal of Cryptosporidium and Giardia spp.

    PubMed

    Bajer, A; Toczylowska, B; Bednarska, M; Sinski, E

    2012-11-01

    Cryptosporidium and Giardia are intestinal parasites of humans and of many other species of animals. Water constitutes an important route of transmission for human infections in both developed and developing countries. In Poland, contamination of water sources with oocysts/cysts is not routinely monitored and scientific research in this field is scarce. Our aim was to compare the contamination of surface and treated water and thus the success of water treatment processes. Water samples (n=94) of between 30 l (surface water) to over 1000 l for tap water, were taken in the period of 2008-2009 using specially constructed equipment with cartridge filtration (Filta-Max; IDEXX, USA). Immunofluorescent assay, and nested polymerase chain reaction were used for the detection of parasites. Cryptosporidium oocysts were found in 85% of surface water and in 59% of raw (intake) water samples. Oocysts were also detected in treated water (16%) but were absent in samples of swimming pool water. The highest mean number of Cryptosporidium oocysts [geometric mean (GM)=61/10 l] was found in samples of rinsing water. Giardia cysts were observed in 61% of surface water samples, in 6% of raw water and in 19% of treated water, with the highest number of cysts noted in rinsing water samples (GM=70 cysts/10 l). Our study highlights the frequent occurrence of parasites in surface waters in Poland and the effectiveness of water treatment for the removal of parasites from drinking water.

  17. The concentration of Cryptosporidium and Giardia in water--the role and importance of recovery efficiency.

    PubMed

    Ongerth, Jerry E

    2013-05-01

    The concentration of Cryptosporidium and of Giardia in surface water is a subject of importance to public health and public water supply. The term concentration is a fundamental property of any water quality parameter having a classical definition as used in chemistry and biology. Analytical methods for measuring the occurrence of Cryptosporidium and Giardia in water find only a fraction of the organisms actually present. This paper collects recently available data to examine the role and importance of recovery efficiency measurement to description of the concentrations of these organisms. Data from Australian sources graphically illustrate the variability of recovery efficiency at individual sites over relatively short time scales. Additional data on replicated recovery measurements establish their reproducibility. The recently released USEPA LT2 data along with those from Australia illustrate the independent variation of Cryptosporidium and Giardia occurrence and recovery efficiency at individual sampling locations. Calculation of concentration from paired raw numbers and recovery efficiency measurements clearly shows the magnitude and importance of taking recovery into account in expressing the concentration of these organisms.

  18. Cryptosporidium spp. in drinking water. Samples from rural sites in Switzerland.

    PubMed

    Füchslin, Hans Peter; Kötzsch, Stefan; Egli, Thomas

    2012-10-04

    In most rural areas and small communities in Switzerland the drinking water is supplied to the consumers after a minimum or even no treatment at all. However, it is just in these areas where drinking water from sources of agricultural activities can be contaminated by liquid manure and faeces of pasturing animals. The Swiss drinking water regulations are limited to the monitoring of E. coli, Enterococcus spp. and total plate counts only. Hence, resistant pathogens, as for example Cryptosporidium spp., remain unnoticed. During a drinking water survey, which lasted from June 2003 to December 2004, water samples were collected from 3 selected rural sites in Switzerland. The drinking water was investigated for Cryptosporidium spp., E. coli, Enterococcus spp., Clostridium perfringens and other parameters. In all samples oocysts of Cryptosporidium spp. were detected at elevated concentrations of up to 0.18 oocysts/l. Between 28% and 75% of the oocysts were found to be vital by the excystation method. Sampled oocysts collected from the three sites were subjected to genotyping and in one case the isolate was found to belong to the genotype of C. parvum. No evidence for increased incidents of diarrhoea in the past years was noted by local authorities.

  19. Source Tracking and Succession of Kimchi Lactic Acid Bacteria during Fermentation.

    PubMed

    Lee, Se Hee; Jung, Ji Young; Jeon, Che Ok

    2015-08-01

    This study aimed at evaluating raw materials as potential lactic acid bacteria (LAB) sources for kimchi fermentation and investigating LAB successions during fermentation. The bacterial abundances and communities of five different sets of raw materials were investigated using plate-counting and pyrosequencing. LAB were found to be highly abundant in all garlic samples, suggesting that garlic may be a major LAB source for kimchi fermentation. LAB were observed in three and two out of five ginger and leek samples, respectively, indicating that they can also be potential important LAB sources. LAB were identified in only one cabbage sample with low abundance, suggesting that cabbage may not be an important LAB source. Bacterial successions during fermentation in the five kimchi samples were investigated by community analysis using pyrosequencing. LAB communities in initial kimchi were similar to the combined LAB communities of individual raw materials, suggesting that kimchi LAB were derived from their raw materials. LAB community analyses showed that species in the genera Leuconostoc, Lactobacillus, and Weissella were key players in kimchi fermentation, but their successions during fermentation varied with the species, indicating that members of the key genera may have different acid tolerance or growth competitiveness depending on their respective species.

  20. ENVIRONMENTAL MONITORING OF POLLUTED WATERS USING SOURCE TRACKING MOLECULAR TOOLS: LESSONS LEARNED AND FUTURE NEEDS

    EPA Science Inventory

    Different approaches have been used to identify fecal pollution sources in water samples. Early on, the fecal coliforms - fecal streptococci ratio was proposed as a method that could discriminate between human and animal contamination. Several studies showed that the latter appro...

  1. Assessment of Equine Fecal Contamination: The Search for Alternative Bacterial Source-tracking Targets

    EPA Science Inventory

    16S rDNA clone libraries were evaluated for detection of fecal source-identifying bacteria from a collapsed equine manure pile. Libraries were constructed using universal eubacterial primers and Bacteroides-Prevotella group-specific primers. Eubacterial sequences indicat...

  2. DEVELOPMENT OF HOST-SPECIFIC METAGENOMIC MARKERS FOR MICROBIAL SOURCE TRACKING USING A NOVEL METAGENOMIC APPROACH

    EPA Science Inventory

    Fecal contamination of source water has always been an important issue to the drinking water industry. Improper disposal of animal waste, leaky septic tanks, storm runoff, and the abundance of wildlife in natural water systems can all be responsible for the spread of enteric path...

  3. Towards establishing a human fecal contamination index in microbial source tracking

    EPA Science Inventory

    There have been significant advances in development of PCR-based methods to detect source associated DNA sequences (markers), but method evaluation has focused on performance with individual challenge samples. Little attention has been given to integration of multiple samples fro...

  4. Stable Isotope Mixing Models as a Tool for Tracking Sources of Water and Water Pollutants

    EPA Science Inventory

    One goal of monitoring pollutants is to be able to trace the pollutant to its source. Here we review how mixing models using stable isotope information on water and water pollutants can help accomplish this goal. A number of elements exist in multiple stable (non-radioactive) i...

  5. Tracking the concentration of heterotrophic plate count bacteria from the source to the consumer's tap.

    PubMed

    Pepper, I L; Rusin, P; Quintanar, D R; Haney, C; Josephson, K L; Gerba, C P

    2004-05-01

    The goal of this project was to quantify the concentration of heterotrophic plate count (HPC) bacteria within water reaching consumer's taps, and from the sources used by a major utility serving the City of Tucson, AZ. With this information, the amounts and sources of HPC bacteria consumed at the tap could be determined. Samples of water were collected on a monthly basis from two well fields, the CAVSARP recovery well field and Southern Avra Valley well field which serves as one of the groundwater sources for Tucson, AZ, and the distribution system which serves the same homes from which tap water was also tested. The average concentration of HPC in source waters within Southern Avra Valley Wells was 56 CFU/ml (range 1-1995/ml). From the CAVSARP recovery well field, corresponding values were 38 CFU/ml (1 to 502 CFU/ml). Unblended groundwater in the chlorinated distribution system averaged 22 CFU/ml (range 1-794). Blended water at the chlorinated distribution site averaged 47 CFU/ml (range 10-158). There was a major shift in the percentage of gram negative to gram-positive bacteria from the wells to the distribution system, to the tap. In the surface CAP source water, 76% of the bacteria were gram-negative compared to 27% gram-negative in the CAVSARP recovery wells. In contrast, Avra Valley wells contained 17% gram-negative bacteria. In both the Tucson groundwater distribution sites and blended distribution sites, the corresponding number of gram negative bacteria was 12%. Finally at the tap, only 0.2% of the bacteria were gram-negative. The average number of bacteria in household taps averaged 3072 HPC/ml and was equal or greater than 500 ml 68% of the time. This study shows that the number of HPC bacteria increases dramatically from the distribution system to the consumers tap. Thus, the major source of bacteria ingested by the average consumer in Tucson originates from bacteria within the household distribution system or the household tap, rather than from source

  6. Tracking the sources and sinks of local marine debris in Hawai'i.

    PubMed

    Carson, Henry S; Lamson, Megan R; Nakashima, Davis; Toloumu, Derek; Hafner, Jan; Maximenko, Nikolai; McDermid, Karla J

    2013-03-01

    Plastic pollution has biological, chemical, and physical effects on marine environments and economic effects on coastal communities. These effects are acute on southeastern Hawai'i Island, where volunteers remove 16 metric tons of debris annually from a 15 km coastline. Although the majority is foreign-origin, a portion is locally-generated. We used floating debris-retention booms in two urban waterways to measure the input of debris from Hilo, the island's largest community, and released wooden drifters in nearby coastal waters to track the fate of that debris. In 205 days, 30 kilograms of debris (73.6% plastic) were retained from two watersheds comprising 10.2% of Hilo's developed land area. Of 851 wooden drifters released offshore of Hilo in four events, 23.3% were recovered locally, 1.4% at distant locations, and 6.5% on other islands. Comparisons with modeled surface currents and wind were mixed, indicating the importance of nearshore and tidal dynamics not included in the model. This study demonstrated that local pollutants can be retained nearby, contribute to the island's debris-accumulation area, and quickly contaminate other islands.

  7. Policy, practice and decision making for zoonotic disease management: water and Cryptosporidium.

    PubMed

    Austin, Zoë; Alcock, Ruth E; Christley, Robert M; Haygarth, Philip M; Heathwaite, A Louise; Latham, Sophia M; Mort, Maggie; Oliver, David M; Pickup, Roger; Wastling, Jonathan M; Wynne, Brian

    2012-04-01

    Decision making for zoonotic disease management should be based on many forms of appropriate data and sources of evidence. However, the criteria and timing for policy response and the resulting management decisions are often altered when a disease outbreak occurs and captures full media attention. In the case of waterborne disease, such as the robust protozoa, Cryptosporidium spp, exposure can cause significant human health risks and preventing exposure by maintaining high standards of biological and chemical water quality remains a priority for water companies in the UK. Little has been documented on how knowledge and information is translated between the many stakeholders involved in the management of Cryptosporidium, which is surprising given the different drivers that have shaped management decisions. Such information, coupled with the uncertainties that surround these data is essential for improving future management strategies that minimise disease outbreaks. Here, we examine the interplay between scientific information, the media, and emergent government and company policies to examine these issues using qualitative and quantitative data relating to Cryptosporidium management decisions by a water company in the North West of England. Our results show that political and media influences are powerful drivers of management decisions if fuelled by high profile outbreaks. Furthermore, the strength of the scientific evidence is often constrained by uncertainties in the data, and in the way knowledge is translated between policy levels during established risk management procedures. In particular, under or over-estimating risk during risk assessment procedures together with uncertainty regarding risk factors within the wider environment, was found to restrict the knowledge-base for decision-making in Cryptosporidium management. Our findings highlight some key current and future challenges facing the management of such diseases that are widely applicable to other

  8. Do oxygen stable isotopes track precipitation moisture source in vascular plant dominated peatlands?

    NASA Astrophysics Data System (ADS)

    Charman, D.; Amesbury, M. J.; Newnham, R.; Loader, N.; Goodrich, J. P.; Gallego-Sala, A. V.; Royles, J.; Keller, E. D.; Baisden, W. T.

    2014-12-01

    Variations in the isotopic composition of precipitation are determined by fractionation processes which occur during temperature and humidity dependent phase changes associated with evaporation and condensation. Oxygen stable isotope ratios have therefore been frequently used as a source of palaeoclimate data from a variety of proxy archives. Exploitation of this record from ombrotrophic peatlands, where the source water used in cellulose synthesis is derived solely from precipitation, has been mostly limited to Northern Hemisphere Sphagnum-dominated bogs, with limited application in the Southern Hemisphere (SH) or in peatlands dominated by vascular plants. Throughout New Zealand (NZ), the preserved root matrix of the restionaceous wire rush (Empodisma spp.) forms deep peat deposits. NZ provides an ideal location to undertake empirical research into oxygen isotope fractionation in vascular peatlands because sites are ideally suited to single taxon analysis, preserve potentially high resolution full Holocene palaeoclimate records and are situated in the climatically sensitive SH mid-latitudes. Crucially, large gradients exist in the mean isotopic composition of precipitation across NZ, caused primarily by the relative influence of different climate modes. We test the capacity for δ18O analysis of Empodisma alpha cellulose from ombrotrophic restiad peatlands in NZ to provide a methodology for developing palaeoclimate records. We took surface plant, water and precipitation samples over spatial (six sites spanning >10° latitude) and temporal (monthly measurements over one year) gradients. We found a strong link between the isotopic compositions of surface root water, the most likely source water for plant growth, and precipitation in both datasets. Back-trajectory modelling of precipitation moisture source for rain days prior to sampling showed clear seasonality in the temporal data that was reflected in surface root water. The link between source water and plant

  9. Molecular Identification of Cryptosporidium Species from Pet Snakes in Thailand

    PubMed Central

    Yimming, Benjarat; Pattanatanang, Khampee; Sanyathitiseree, Pornchai; Inpankaew, Tawin; Kamyingkird, Ketsarin; Pinyopanuwat, Nongnuch; Chimnoi, Wissanuwat; Phasuk, Jumnongjit

    2016-01-01

    Cryptosporidium is an important pathogen causing gastrointestinal disease in snakes and is distributed worldwide. The main objectives of this study were to detect and identify Cryptosporidium species in captive snakes from exotic pet shops and snake farms in Thailand. In total, 165 fecal samples were examined from 8 snake species, boa constrictor (Boa constrictor constrictor), corn snake (Elaphe guttata), ball python (Python regius), milk snake (Lampropeltis triangulum), king snake (Lampropeltis getula), rock python (Python sebae), rainbow boa (Epicrates cenchria), and carpet python (Morelia spilota). Cryptosporidium oocysts were examined using the dimethyl sulfoxide (DMSO)-modified acid-fast staining and a molecular method based on nested-PCR, PCR-RFLP analysis, and sequencing amplification of the SSU rRNA gene. DMSO-modified acid-fast staining revealed the presence of Cryptosporidium oocysts in 12 out of 165 (7.3%) samples, whereas PCR produced positive results in 40 (24.2%) samples. Molecular characterization indicated the presence of Cryptosporidium parvum (mouse genotype) as the most common species in 24 samples (60%) from 5 species of snake followed by Cryptosporidium serpentis in 9 samples (22.5%) from 2 species of snake and Cryptosporidium muris in 3 samples (7.5%) from P. regius. PMID:27658593

  10. Molecular Identification of Cryptosporidium Species from Pet Snakes in Thailand.

    PubMed

    Yimming, Benjarat; Pattanatanang, Khampee; Sanyathitiseree, Pornchai; Inpankaew, Tawin; Kamyingkird, Ketsarin; Pinyopanuwat, Nongnuch; Chimnoi, Wissanuwat; Phasuk, Jumnongjit

    2016-08-01

    Cryptosporidium is an important pathogen causing gastrointestinal disease in snakes and is distributed worldwide. The main objectives of this study were to detect and identify Cryptosporidium species in captive snakes from exotic pet shops and snake farms in Thailand. In total, 165 fecal samples were examined from 8 snake species, boa constrictor (Boa constrictor constrictor), corn snake (Elaphe guttata), ball python (Python regius), milk snake (Lampropeltis triangulum), king snake (Lampropeltis getula), rock python (Python sebae), rainbow boa (Epicrates cenchria), and carpet python (Morelia spilota). Cryptosporidium oocysts were examined using the dimethyl sulfoxide (DMSO)-modified acid-fast staining and a molecular method based on nested-PCR, PCR-RFLP analysis, and sequencing amplification of the SSU rRNA gene. DMSO-modified acid-fast staining revealed the presence of Cryptosporidium oocysts in 12 out of 165 (7.3%) samples, whereas PCR produced positive results in 40 (24.2%) samples. Molecular characterization indicated the presence of Cryptosporidium parvum (mouse genotype) as the most common species in 24 samples (60%) from 5 species of snake followed by Cryptosporidium serpentis in 9 samples (22.5%) from 2 species of snake and Cryptosporidium muris in 3 samples (7.5%) from P. regius.

  11. En face projection imaging of the human choroidal layers with tracking SLO and swept source OCT angiography methods

    NASA Astrophysics Data System (ADS)

    Gorczynska, Iwona; Migacz, Justin; Zawadzki, Robert J.; Sudheendran, Narendran; Jian, Yifan; Tiruveedhula, Pavan K.; Roorda, Austin; Werner, John S.

    2015-07-01

    We tested and compared the capability of multiple optical coherence tomography (OCT) angiography methods: phase variance, amplitude decorrelation and speckle variance, with application of the split spectrum technique, to image the choroiretinal complex of the human eye. To test the possibility of OCT imaging stability improvement we utilized a real-time tracking scanning laser ophthalmoscopy (TSLO) system combined with a swept source OCT setup. In addition, we implemented a post- processing volume averaging method for improved angiographic image quality and reduction of motion artifacts. The OCT system operated at the central wavelength of 1040nm to enable sufficient depth penetration into the choroid. Imaging was performed in the eyes of healthy volunteers and patients diagnosed with age-related macular degeneration.

  12. Cryptosporidium spp. and Giardia duodenalis as pathogenic contaminants of water in Galicia, Spain: the need for safe drinking water.

    PubMed

    Castro-Hermida, José Antonio; González-Warleta, Marta; Mezo, Mercedes

    2015-01-01

    The objectives of this cross-sectional study were to detect the presence of Cryptosporidium spp. and Giardia duodenalis in drinking water treatments plants (DWTPs) in Galicia (NW Spain) and to identify which species and genotype of these pathogenic protozoans are present in the water. Samples of untreated water (surface or ground water sources) and of treated drinking water (in total, 254 samples) were collected from 127 DWTPs and analysed by an immunofluorescence antibody test (IFAT) and by PCR. Considering the untreated water samples, Cryptosporidium spp. were detected in 69 samples (54.3%) by IFAT, and DNA of this parasite was detected in 57 samples (44.8%) by PCR, whereas G. duodenalis was detected in 76 samples (59.8%) by IFAT and in 56 samples (44.0%) by PCR. Considering the treated drinking water samples, Cryptosporidium spp. was detected in 52 samples (40.9%) by IFAT, and the parasite DNA was detected in 51 samples (40.1%) by PCR, whereas G. duodenalis was detected in 58 samples (45.6%) by IFAT and in 43 samples (33.8%) by PCR. The percentage viability of the (oo)cysts ranged between 90.0% and 95.0% in all samples analysed. Cryptosporidium andersoni, C. hominis, C. parvum and assemblages A-I, A-II, E of G. duodenalis were identified. The results indicate that Cryptosporidium spp. and G. duodenalis are widespread in the environment and that DWTPs are largely ineffective in reducing/inactivating these pathogens in drinking water destined for human and animal consumption in Galicia. In conclusion, the findings suggest the need for better monitoring of water quality and identification of sources of contamination.

  13. Rotifers ingest oocysts of Cryptosporidium parvum

    USGS Publications Warehouse

    Fayer, R.; Trout, J.M.; Walsh, E.; Cole, R.A.

    2000-01-01

    Six genera of rotifers including Philodina, Monostyla, Epiphanes, Euchlanis, Brachionus, and Asplanchna were exposed to oocysts of Cryptosporidium parvum cleaned of fecal debris. Unstained oocysts and those stained with fluorescein-conjugated monoclonal antibody were added to suspensions of viable rotifers and were examined by phase-contrast, differential interference contrast, and fluorescence microscopy. Rotifers of all six genera were observed ingesting oocysts. A maximum of 25 oocysts was observed in the stomachs of Euchlanis and Brachionus. Euchlanis and Epiphanes were observed excreting boluses containing up to eight oocysts. It was not determined whether rotifers digested or otherwise rendered oocysts nonviable.

  14. Persistence and microbial source tracking of Escherichia coli at a swimming beach at Lake of the Ozarks State Park, Missouri

    USGS Publications Warehouse

    Wilson, Jordan L.; Schumacher, John G.; Burken, Joel G.

    2016-01-01

    The Missouri Department of Natural Resources (MDNR) has closed or posted advisories at public beaches at Lake of the Ozarks State Park in Missouri because of Escherichia coli (E. coli) concentration exceedances in recent years. Spatial and temporal patterns of E. coliconcentrations, microbial source tracking, novel sampling techniques, and beach-use patterns were studied during the 2012 recreational season to identify possible sources, origins, and occurrence of E. coli contamination at Grand Glaize Beach (GGB). Results indicate an important source of E. coli contamination at GGB was E. coli released into the water column by bathers resuspending avian-contaminated sediments, especially during high-use days early in the recreational season. Escherichia coli concentrations in water, sediment, and resuspended sediment samples all decreased throughout the recreational season likely because of decreasing lake levels resulting in sampling locations receding away from the initial spring shoreline as well as natural decay and physical transport out of the cove. Weekly MDNR beach monitoring, based solely on E. coli concentrations, at GGB during this study inaccurately predicted E. coli exceedances, especially on weekends and holidays. Interestingly, E. coli of human origin were measured at concentrations indicative of raw sewage in runoff from an excavation of a nearby abandoned septic tank that had not been used for nearly two years.

  15. Tracking Type III Radio Burst Sources in the Solar Corona by Heliographic Means

    NASA Astrophysics Data System (ADS)

    Koval, A. A.; Stanislavsky, A. A.; Konovalenko, A. A.; Volvach, Ya. S.

    We present the preliminary results of heliographic measurements of solar type III radio bursts in the low-frequency range (16.5-33 MHz) using the UTR-2 radio heliograph. The radio astronomy tools permit us to obtain two-dimensional spatial structures of burst sources in dependence of frequency and time. Each heliogram consists of 40 pixels (beams) as a result of the serial sweep in UV-plane wherein signals of each beam are recorded in a dynamic spectrum with both high temporal (˜ 2.482 ms) and top spectral (˜ 4 kHz) resolutions. The rate of output heliograph is one image per 3 seconds. Over a session in April, 2013 many type III radio and IIIb-III bursts were observed. On the heliograms the source motion direction in the upper corona is clearly detectable. The heliogram features are discussed.

  16. Real-Time Localization of Moving Dipole Sources for Tracking Multiple Free-Swimming Weakly Electric Fish

    PubMed Central

    Jun, James Jaeyoon; Longtin, André; Maler, Leonard

    2013-01-01

    In order to survive, animals must quickly and accurately locate prey, predators, and conspecifics using the signals they generate. The signal source location can be estimated using multiple detectors and the inverse relationship between the received signal intensity (RSI) and the distance, but difficulty of the source localization increases if there is an additional dependence on the orientation of a signal source. In such cases, the signal source could be approximated as an ideal dipole for simplification. Based on a theoretical model, the RSI can be directly predicted from a known dipole location; but estimating a dipole location from RSIs has no direct analytical solution. Here, we propose an efficient solution to the dipole localization problem by using a lookup table (LUT) to store RSIs predicted by our theoretically derived dipole model at many possible dipole positions and orientations. For a given set of RSIs measured at multiple detectors, our algorithm found a dipole location having the closest matching normalized RSIs from the LUT, and further refined the location at higher resolution. Studying the natural behavior of weakly electric fish (WEF) requires efficiently computing their location and the temporal pattern of their electric signals over extended periods. Our dipole localization method was successfully applied to track single or multiple freely swimming WEF in shallow water in real-time, as each fish could be closely approximated by an ideal current dipole in two dimensions. Our optimized search algorithm found the animal’s positions, orientations, and tail-bending angles quickly and accurately under various conditions, without the need for calibrating individual-specific parameters. Our dipole localization method is directly applicable to studying the role of active sensing during spatial navigation, or social interactions between multiple WEF. Furthermore, our method could be extended to other application areas involving dipole source

  17. Real-Time Localization of Moving Dipole Sources for Tracking Multiple Free-Swimming Weakly Electric Fish.

    PubMed

    Jun, James Jaeyoon; Longtin, André; Maler, Leonard

    2013-01-01

    In order to survive, animals must quickly and accurately locate prey, predators, and conspecifics using the signals they generate. The signal source location can be estimated using multiple detectors and the inverse relationship between the received signal intensity (RSI) and the distance, but difficulty of the source localization increases if there is an additional dependence on the orientation of a signal source. In such cases, the signal source could be approximated as an ideal dipole for simplification. Based on a theoretical model, the RSI can be directly predicted from a known dipole location; but estimating a dipole location from RSIs has no direct analytical solution. Here, we propose an efficient solution to the dipole localization problem by using a lookup table (LUT) to store RSIs predicted by our theoretically derived dipole model at many possible dipole positions and orientations. For a given set of RSIs measured at multiple detectors, our algorithm found a dipole location having the closest matching normalized RSIs from the LUT, and further refined the location at higher resolution. Studying the natural behavior of weakly electric fish (WEF) requires efficiently computing their location and the temporal pattern of their electric signals over extended periods. Our dipole localization method was successfully applied to track single or multiple freely swimming WEF in shallow water in real-time, as each fish could be closely approximated by an ideal current dipole in two dimensions. Our optimized search algorithm found the animal's positions, orientations, and tail-bending angles quickly and accurately under various conditions, without the need for calibrating individual-specific parameters. Our dipole localization method is directly applicable to studying the role of active sensing during spatial navigation, or social interactions between multiple WEF. Furthermore, our method could be extended to other application areas involving dipole source

  18. Decay kinetics of microbial source tracking (MST) markers and human adenovirus under the effects of sunlight and salinity.

    PubMed

    L, Liang; S G, Goh; K Y H, Gin

    2017-01-01

    Artificial seawater and freshwater microcosms inoculated with raw sewage were set up to compare the persistence of microbial source tracking (MST) markers (i.e. Bacteroides thetaiotaomicron (B. theta), Methanobrevibacter smithii (M. smithii), human polyomaviruses JC and BK (HPyVs)) and human adenoviruses under different sunlight intensity and salinity. PMA pretreatment successfully eliminated the false-positive detection of dead bacterial cells in the model-development experiment. The results were then validated using real environmental matrices in microcosms inoculated with raw sewage. The genome concentrations of the targets followed a first-order decay pattern with 90% reduction of the initial amounts in <5days for both artificial and natural surface waters. Decay rate constant (k1) were developed microorganisms in artificial water matrices. Due to the different water environment conditions, improved decay rates (k2) incorporated with sunlight, TSS and TOC adjustment coefficients were used for validation of the natural water matrices. Based on the predictive squared correlation coefficient (Q(2)F) and root-mean-square error (RMSE) validation criteria, the improved k2 were able to provide better prediction on the survival of target microorganisms in environmental surface waters (Q(2)F>0.6 and RMSE ranged from 0.05 to 1.81). For microbial source tracking purposes, HPyVs are suggested to be better MST markers in freshwater, while B. theta is recommended for seawater based on the decay models developed in this study. The targeted DNA of M. smithii should only be used to indicate recent human faecal pollution in surface waters due to their faster decay than human adenoviruses.

  19. Track-a-worm, an open-source system for quantitative assessment of C. elegans locomotory and bending behavior.

    PubMed

    Wang, Sijie Jason; Wang, Zhao-Wen

    2013-01-01

    A major challenge of neuroscience is to understand the circuit and gene bases of behavior. C. elegans is commonly used as a model system to investigate how various gene products function at specific tissue, cellular, and synaptic foci to produce complicated locomotory and bending behavior. The investigation generally requires quantitative behavioral analyses using an automated single-worm tracker, which constantly records and analyzes the position and body shape of a freely moving worm at a high magnification. Many single-worm trackers have been developed to meet lab-specific needs, but none has been widely implemented for various reasons, such as hardware difficult to assemble, and software lacking sufficient functionality, having closed source code, or using a programming language that is not broadly accessible. The lack of a versatile system convenient for wide implementation makes data comparisons difficult and compels other labs to develop new worm trackers. Here we describe Track-A-Worm, a system rich in functionality, open in source code, and easy to use. The system includes plug-and-play hardware (a stereomicroscope, a digital camera and a motorized stage), custom software written to run with Matlab in Windows 7, and a detailed user manual. Grayscale images are automatically converted to binary images followed by head identification and placement of 13 markers along a deduced spline. The software can extract and quantify a variety of parameters, including distance traveled, average speed, distance/time/speed of forward and backward locomotion, frequency and amplitude of dominant bends, overall bending activities measured as root mean square, and sum of all bends. It also plots worm travel path, bend trace, and bend frequency spectrum. All functionality is performed through graphical user interfaces and data is exported to clearly-annotated and documented Excel files. These features make Track-A-Worm a good candidate for implementation in other labs.

  20. Physico-chemical characterization and source tracking of black carbon at a suburban site in Beijing.

    PubMed

    Wang, Hailin; Nie, Lei; Liu, Dan; Gao, Meiping; Wang, Minyan; Hao, Zhengping

    2015-07-01

    Particles from ambient air and combustion sources including vehicle emission, coal combustion and biomass burning were collected and chemically pretreated with the purpose of obtaining isolated BC (black carbon) samples. TEM (transmission electron microscopy) results indicate that BC from combustion sources shows various patterns, and airborne BC appears spherical and about 50 nm in diameter with a homogeneous surface and turbostratic structure. The BET (Barrett-Emmett-Teller) results suggest that the surface areas of these BC particles fall in the range of 3-23 m2/g, with a total pore volume of 0.03-0.05 cm3/g and a mean pore diameter of 7-53 nm. The nitrogen adsorption-desorption isotherms are indicative of the accumulation mode and uniform pore size. O2-TPO (temperature programmed oxidation) profiles suggest that the airborne BC oxidation could be classified as the oxidation of amorphous carbon, which falls in the range of 406-490°C with peaks at 418, 423 and 475°C, respectively. Generally, the BC characteristics and source analysis suggest that airborne BC most likely comes from diesel vehicle emission at this site.

  1. Probabilistic analysis showing that a combination of bacteroides and methanobrevibacter source tracking markers is effective for identifying waters contaminated by human fecal pollution

    USGS Publications Warehouse

    Johnston, Christopher; Byappanahalli, Muruleedhara N.; Gibson, Jacqueline MacDonald; Ufnar, Jennifer A.; Whitman, Richard L.; Stewart, Jill R.

    2013-01-01

    Microbial source tracking assays to identify sources of waterborne contamination typically target genetic markers of host-specific microorganisms. However, no bacterial marker has been shown to be 100% host-specific, and cross-reactivity has been noted in studies evaluating known source samples. Using 485 challenge samples from 20 different human and animal fecal sources, this study evaluated microbial source tracking markers including the Bacteroides HF183 16S rRNA, M. smithii nifH, and Enterococcus esp gene targets that have been proposed as potential indicators of human fecal contamination. Bayes' Theorem was used to calculate the conditional probability that these markers or a combination of markers can correctly identify human sources of fecal pollution. All three human-associated markers were detected in 100% of the sewage samples analyzed. Bacteroides HF183 was the most effective marker for determining whether contamination was specifically from a human source, and greater than 98% certainty that contamination was from a human source was shown when both Bacteroides HF183 and M. smithii nifH markers were present. A high degree of certainty was attained even in cases where the prior probability of human fecal contamination was as low as 8.5%. The combination of Bacteroides HF183 and M. smithii nifH source tracking markers can help identify surface waters impacted by human fecal contamination, information useful for prioritizing restoration activities or assessing health risks from exposure to contaminated waters.

  2. UV Inactivation of Cryptosporidium hominis as Measured in Cell Culture

    PubMed Central

    Johnson, Anne M.; Linden, Karl; Ciociola, Kristina M.; De Leon, Ricardo; Widmer, Giovanni; Rochelle, Paul A.

    2005-01-01

    The Cryptosporidium spp. UV disinfection studies conducted to date have used Cryptosporidium parvum oocysts. However, Cryptosporidium hominis predominates in human cryptosporidiosis infections, so there is a critical need to assess the efficacy of UV disinfection of C. hominis. This study utilized cell culture-based methods to demonstrate that C. hominis oocysts displayed similar levels of infectivity and had the same sensitivity to UV light as C. parvum. Therefore, the water industry can be confident about extrapolating C. parvum UV disinfection data to C. hominis oocysts. PMID:15870378

  3. Cryptosporidium varanii infection in leopard geckos (Eublepharis macularius) in Argentina.

    PubMed

    Dellarupe, A; Unzaga, J M; Moré, G; Kienast, M; Larsen, A; Stiebel, C; Rambeaud, M; Venturini, M C

    2016-01-01

    Cryptosporidiosis is observed in reptiles with high morbidity and considerable mortality. The objective of this study was to achieve the molecular identification of Cryptosporidium spp. in pet leopard geckos (Eublepharis macularius) from a breeder colony in Buenos Aires, Argentina. Oocysts comparable to those of Cryptosporidium spp. were detected in three geckos with a history of diarrhea, anorexia and cachexia. Molecular identification methods confirmed the presence of Cryptosporidium varanii (syn. C. saurophilum). This agent was considered to be the primary cause of the observed clinical disease. This is the first description of C. varanii infection in pet reptiles in Argentina.

  4. Cryptosporidium varanii infection in leopard geckos (Eublepharis macularius) in Argentina

    PubMed Central

    Dellarupe, A.; Unzaga, J.M.; Moré, G.; Kienast, M.; Larsen, A.; Stiebel, C.; Rambeaud, M.; Venturini, M.C.

    2016-01-01

    Cryptosporidiosis is observed in reptiles with high morbidity and considerable mortality. The objective of this study was to achieve the molecular identification of Cryptosporidium spp. in pet leopard geckos (Eublepharis macularius) from a breeder colony in Buenos Aires, Argentina. Oocysts comparable to those of Cryptosporidium spp. were detected in three geckos with a history of diarrhea, anorexia and cachexia. Molecular identification methods confirmed the presence of Cryptosporidium varanii (syn. C. saurophilum). This agent was considered to be the primary cause of the observed clinical disease. This is the first description of C. varanii infection in pet reptiles in Argentina. PMID:27419102

  5. Tracking Nitrogen Sources in Urban Stormwater Runoff from an Urban Residential Neighborhood

    NASA Astrophysics Data System (ADS)

    Toor, G.

    2015-12-01

    The magnitude of nitrogen (N) flux from urban residential neighborhoods to water bodies is not known. We present a case study of N evolution in urban stormwater runoff from a residential neighborhood located in Florida where most runoff is generated by excess rainfall. This rainfall occurs during the 4-month wet season (June-September) that receives ~60 to 70% of 125 cm annual rainfall. The outlet pipe draining the residential neighborhood in Florida was instrumented with ISCO sampler. Stormwater runoff samples were collected every 5-minute following variable storm durations and frequencies during 2014 wet season. Mean concentration of total N in stormwater runoff ranged from 1 to 3 mg L-1; of which, nitrate-N was <25% and organic N was >60% in most storm events; suggesting different processes controlling N release and transport during variable storm events. Stormwater samples were analyzed for N and oxygen (O) isotopes of nitrate along with hydrogen (H) and O in water to understand the sources of N and water. This presentation will discuss our ongoing research aimed at better understanding the sources, processes, and unraveling the unknowns and uncertainty of N transport from urban coastal systems

  6. Use of Spatial Sampling and Microbial Source-Tracking Tools for Understanding Fecal Contamination at Two Lake Erie Beaches

    USGS Publications Warehouse

    Francy, Donna S.; Bertke, Erin E.; Finnegan, Dennis P.; Kephart, Christopher M.; Sheets, Rodney A.; Rhoades, John; Stumpe, Lester

    2006-01-01

    Source-tracking tools were used to identify potential sources of fecal contamination at two Lake Erie bathing beaches: an urban beach (Edgewater in Cleveland, Ohio) and a beach in a small city (Lakeshore in Ashtabula, Ohio). These tools included identifying spatial patterns of Escherichia coli (E. coli) concentrations in each area, determining weather patterns that caused elevated E. coli, and applying microbial source tracking (MST) techniques to specific sites. Three MST methods were used during this study: multiple antibiotic resistance (MAR) indexing of E. coli isolates and the presence of human-specific genetic markers within two types of bacteria, the genus Bacteroides and the species Enterococcus faecium. At Edgewater, sampling for E. coli was done during 2003-05 at bathing-area sites, at nearshore lake sites, and in shallow ground water in foreshore and backshore areas. Spatial sampling at nearshore lake sites showed that fecal contamination was most likely of local origin; E. coli concentrations near the mouths of rivers and outfalls remote to the beach were elevated (greater than 235 colony-forming units per 100 milliliters (CFU/100 mL)) but decreased along transport pathways to the beach. In addition, E. coli concentrations were generally highest in bathing-area samples collected at 1- and 2-foot water depths, midrange at 3-foot depths, and lowest in nearshore lake samples typically collected 150 feet from the shoreline. Elevated E. coli concentrations at bathing-area sites were generally associated with increased wave heights and rainfall, but not always. E. coli concentrations were often elevated in shallow ground-water samples, especially in samples collected less than 10 feet from the edge of water (near foreshore area). The interaction of shallow ground water and waves may be a mechanism of E. coli storage and accumulation in foreshore sands. Infiltration of bird feces through sand with surface water from rainfall and high waves may be concentrating

  7. Ultrafiltration and Microarray for Detection of Microbial Source Tracking Marker and Pathogen Genes in Riverine and Marine Systems.

    PubMed

    Li, Xiang; Harwood, Valerie J; Nayak, Bina; Weidhaas, Jennifer L

    2016-01-04

    Pathogen identification and microbial source tracking (MST) to identify sources of fecal pollution improve evaluation of water quality. They contribute to improved assessment of human health risks and remediation of pollution sources. An MST microarray was used to simultaneously detect genes for multiple pathogens and indicators of fecal pollution in freshwater, marine water, sewage-contaminated freshwater and marine water, and treated wastewater. Dead-end ultrafiltration (DEUF) was used to concentrate organisms from water samples, yielding a recovery efficiency of >95% for Escherichia coli and human polyomavirus. Whole-genome amplification (WGA) increased gene copies from ultrafiltered samples and increased the sensitivity of the microarray. Viruses (adenovirus, bocavirus, hepatitis A virus, and human polyomaviruses) were detected in sewage-contaminated samples. Pathogens such as Legionella pneumophila, Shigella flexneri, and Campylobacter fetus were detected along with genes conferring resistance to aminoglycosides, beta-lactams, and tetracycline. Nonmetric dimensional analysis of MST marker genes grouped sewage-spiked freshwater and marine samples with sewage and apart from other fecal sources. The sensitivity (percent true positives) of the microarray probes for gene targets anticipated in sewage was 51 to 57% and was lower than the specificity (percent true negatives; 79 to 81%). A linear relationship between gene copies determined by quantitative PCR and microarray fluorescence was found, indicating the semiquantitative nature of the MST microarray. These results indicate that ultrafiltration coupled with WGA provides sufficient nucleic acids for detection of viruses, bacteria, protozoa, and antibiotic resistance genes by the microarray in applications ranging from beach monitoring to risk assessment.

  8. Ultrafiltration and Microarray for Detection of Microbial Source Tracking Marker and Pathogen Genes in Riverine and Marine Systems

    PubMed Central

    Li, Xiang; Harwood, Valerie J.; Nayak, Bina

    2016-01-01

    Pathogen identification and microbial source tracking (MST) to identify sources of fecal pollution improve evaluation of water quality. They contribute to improved assessment of human health risks and remediation of pollution sources. An MST microarray was used to simultaneously detect genes for multiple pathogens and indicators of fecal pollution in freshwater, marine water, sewage-contaminated freshwater and marine water, and treated wastewater. Dead-end ultrafiltration (DEUF) was used to concentrate organisms from water samples, yielding a recovery efficiency of >95% for Escherichia coli and human polyomavirus. Whole-genome amplification (WGA) increased gene copies from ultrafiltered samples and increased the sensitivity of the microarray. Viruses (adenovirus, bocavirus, hepatitis A virus, and human polyomaviruses) were detected in sewage-contaminated samples. Pathogens such as Legionella pneumophila, Shigella flexneri, and Campylobacter fetus were detected along with genes conferring resistance to aminoglycosides, beta-lactams, and tetracycline. Nonmetric dimensional analysis of MST marker genes grouped sewage-spiked freshwater and marine samples with sewage and apart from other fecal sources. The sensitivity (percent true positives) of the microarray probes for gene targets anticipated in sewage was 51 to 57% and was lower than the specificity (percent true negatives; 79 to 81%). A linear relationship between gene copies determined by quantitative PCR and microarray fluorescence was found, indicating the semiquantitative nature of the MST microarray. These results indicate that ultrafiltration coupled with WGA provides sufficient nucleic acids for detection of viruses, bacteria, protozoa, and antibiotic resistance genes by the microarray in applications ranging from beach monitoring to risk assessment. PMID:26729716

  9. Cryptosporidium parvum Infection Following Contact with Livestock

    PubMed Central

    Suler, Denis; Mullins, David; Rudge, Travis; Ashurst, John

    2016-01-01

    Context: Scours, or calf diarrhea, is an infectious gastrointestinal disease commonly found in the calves of dairy farms. It primarily presents with diarrhea that can be life threatening to the animal and is also contagious and threatening to the other livestock. Cryptosporidium is one of the major causes of scours and can be transmitted to humans via fecal-oral route, resulting in diarrheal illnesses. Cryptosporidiosis infection usually occurs as a waterborne outbreak with the potential to affect many people at once. Case Report: We report a case of a 24-year-old female farmer who presented to the emergency department with diarrhea after taking care of ill cattle with similar symptoms. Fecal cultures were positive for Cryptosporidium parvum. Given the patient was immunocompetent, no further treatment was warranted. Conclusion: Confirmed cases should be reported, however, treatment is only recommended in children and immunocompromised adults. Clinicians should educate patients on the importance of proper hygiene and handling techniques in order to decrease transmission and recurrence of the protozoan infection. PMID:27583243

  10. Cryptosporidium infection in solid organ transplantation

    PubMed Central

    Florescu, Diana F; Sandkovsky, Uriel

    2016-01-01

    Diarrhea is a common complication in solid organ transplant (SOT) recipients and may be attributed to immunosuppressive drugs or infectious organisms such as bacteria, viruses or parasites. Cryptosporidium usually causes self-limited diarrhea in immunocompetent hosts. Although it is estimated that cryptosporidium is involved in about 12% of cases of infectious diarrhea in developing countries and causes approximately 748000 cases each year in the United States, it is still an under recognized and important cause of infectious diarrhea in SOT recipients. It may run a protracted course with severe diarrhea, fluid and electrolyte depletion and potential for organ failure. Although diagnostic methodologies have improved significantly, allowing for fast and accurate identification of the parasite, treatment of the disease is difficult because antiparasitic drugs have modest activity at best. Current management includes fluid and electrolyte replacement, reduction of immunosuppression and single therapy with Nitazoxanide or combination therapy with Nitazoxanide and other drugs. Future drug and vaccine development may add to the currently poor armamentarium to manage the disease. The current review highlights key epidemiological, diagnostic and management issues in the SOT population. PMID:27683627

  11. Association of fecal indicator bacteria with human viruses and microbial source tracking markers at coastal beaches impacted by nonpoint source pollution.

    PubMed

    McQuaig, Shannon; Griffith, John; Harwood, Valerie J

    2012-09-01

    Water quality was assessed at two marine beaches in California by measuring the concentrations of culturable fecal indicator bacteria (FIB) and by library-independent microbial source tracking (MST) methods targeting markers of human-associated microbes (human polyomavirus [HPyV] PCR and quantitative PCR, Methanobrevibacter smithii PCR, and Bacteroides sp. strain HF183 PCR) and a human pathogen (adenovirus by nested PCR). FIB levels periodically exceeded regulatory thresholds at Doheny and Avalon Beaches for enterococci (28.5% and 31.7% of samples, respectively) and fecal coliforms (20% and 5.8%, respectively). Adenoviruses were detected at four of five sites at Doheny Beach and were correlated with detection of HPyVs and human Bacteroides HF183; however, adenoviruses were not detected at Avalon Beach. The most frequently detected human source marker at both beaches was Bacteroides HF183, which was detected in 27% of samples. Correlations between FIBs and human markers were much more frequent at Doheny Beach than at Avalon Beach; e.g., adenovirus was correlated with HPyVs and HF183. Human sewage markers and adenoviruses were routinely detected in samples meeting FIB regulatory standards. The toolbox approach of FIB measurement coupled with analysis of several MST markers targeting human pathogens used here demonstrated that human sewage is at least partly responsible for the degradation of water quality, particularly at Doheny Beach, and resulted in a more definitive assessment of recreational water quality and human health risk than reliance on FIB concentrations alone could have provided.

  12. Fission-track Evidence for the Source of Brahmaputra River Sands Within the Eastern Himalayan Syntaxis: a Large Flux from a Tiny Source

    NASA Astrophysics Data System (ADS)

    Stewart, R. J.; Hallet, B.; Zeitler, P. K.

    2006-12-01

    We present new zircon and apatite fission-track results from river sands of the Brahmaputra system; they complement our prior results and add to diverse lines of evidence indicating that erosion in the core of the eastern Himalayan syntaxis has been and is exceptionally rapid. These new results improve definition of the bedrock source area for very young grains: a source of uncertainty in our original data set stemmed from our key downstream sampling site being at Pashigat, on the floodplain of the Brahmaputra, permitting drainages other than the Tsangpo/Siang (local names for the upper Brahmaputra) from being potential contributors of young grains. One important new sample was collected near Medoc, in the lower reaches of the Tsangpo gorge, allowing us to tightly bracket detrital contributions from this deep gorge through the geologically active Nanche Barwa-Gyala Peri massif, the likely source of very young cooling ages of less than 2 Ma. The second sample was collected from a small river draining the cirque glacier incising the NW side of Namche Barwa. We report analyses of 37 zircon grains and 66 apatite grains from the Medoc sample and 80 zircon grains from the cirque sample. Our new results are as follows (previous results from Pashigat are shown in parentheses). The youngest peak identified by BINOMFIT in detrital zircons from Medoc is 0.6 Ma (0.6 Ma), and significantly, it includes 51% (47%) of the entire sand-sized population. The youngest grains are ~ 0.1 Ma (0.1 Ma), and a significant subset has a peak age of 0.3 to 0.4 Ma (0.4 Ma). The youngest peak in apatite fission-track ages from the same samples is 0.5 Ma (0.4 Ma) and includes 58% (39%) of the grains. Zircons from the Namche Barwa cirque also yield a population of extremely young ages having a number of peaks, the youngest of which is 0.3 Ma and accounts for 35% of the grains; the oldest grain in this entire sample is 3.3 Ma. The age distributions from Medoc and Pasighat are very similar, giving us

  13. Prevalence and distribution of Cryptosporidium and Giardia in wastewater and the surface, drinking and ground waters in the Lower Rhine, Germany.

    PubMed

    Gallas-Lindemann, C; Sotiriadou, I; Plutzer, J; Karanis, P

    2013-01-01

    Samples from different water sources (n = 396) were collected during 2009 and 2011. Wastewater (2-5 l) was purified by aluminium sulphate flocculation. Surface, ground and drinking waters (400-6400 l) were collected by filtration. Cryptosporidium oocysts and Giardia cysts were further concentrated by sucrose centrifugation. (Oo)cysts were identified by IFT (immunofluorescence test), DAPI (4',6-diamidino-2-phenylindole) staining and DICM (difference interference contrast microscopy). Out of 206 wastewater samples, 134 (65·0%) were found to be positive for Giardia cysts and 64 (31·1%) for Cryptosporidium oocysts. Parasite numbers ranged from 0 to 2436 cysts/l and 0 to 1745 oocysts/l. Eight (4·2%) surface and drinking water samples (n = 190) were found to be positive for Giardia cysts (0-56000/100 l), and 18 (9·5%) for Cryptosporidium oocysts (2400/100 l). The purpose of this study was to establish the prevalence and concentrations of Giardia lamblia and Cryptosporidium spp. by detecting (oo)cysts from water samples. This study provides substantial evidence that G. lamblia cysts and Cryptosporidium spp. oocysts are able to enter and circulate in the aquatic environment with negative implications for public health.

  14. Questionable Specificity of Genetic Total Faecal Pollution Markers for Integrated Water Quality Monitoring and Source Tracking

    NASA Astrophysics Data System (ADS)

    Vierheilig, Julia; Reischer, Georg H.; Farnleitner, Andreas H.

    2010-05-01

    Characterisation of microbial faecal hazards in water is a fundamental aspect for target-orientated water resources management to achieve appropriate water quality for various purposes like water supply or agriculture and thus to minimize related health risks. Nowadays the management of water resources increasingly demands detailed knowledge on the extent and the origin of microbial pollution. Cultivation of standard faecal indicator bacteria, which has been used for over a century to test the microbiological water quality, cannot sufficiently meet these challenges. The abundant intestinal bacterial populations are very promising alternative targets for modern faecal indication systems. Numerous assays for the detection of genetic markers targeting source-specific populations of the phylum Bacteroidetes have been developed in recent years. In some cases markers for total faecal pollution were also proposed in order to relate source-specific marker concentrations to general faecal pollution levels. However, microbial populations in intestinal and non-intestinal systems exhibit a dazzling array of diversity and molecular analysis of microbial faecal pollution has been based on a fragmentary puzzle of very limited sequence information. The aim of this study was to test the available qPCR-based methods detecting genetic Bacteroidetes markers for total faecal pollution in terms of their value and specificity as indicators of faecal pollution. We applied the AllBac (Layton et al., 2006) the BacUni (Kildare et al., 2007) and the Bacteroidetes (Dick and Field, 2004) assays on soil DNA samples. Samples were collected in well characterised karst spring catchments in Austria's Eastern Calcareous Alps. They were at various levels of altitude between 800 and 1800 meters above sea level and from several different habitats (woodland, alpine pastures, krummholz). In addition we tried to choose sampling sites representing a presumptive gradient of faecal pollution levels. For

  15. Current and future trends in fecal source tracking and deployment in the Lake Taihu Region of China

    NASA Astrophysics Data System (ADS)

    Hagedorn, Charles; Liang, Xinqiang

    The emerging discipline of microbial and/or chemical source tracking (collectively termed fecal source tracking (FST)) is being used to identify origins of fecal contamination in polluted waters in many countries around the world. FST has developed rapidly because standard methods of measuring contamination in water by enumerating fecal indicator bacteria (FIB) such as fecal coliforms and enterococci do not identify the sources of the contamination. FST is an active area of research and development in both the academic and private sectors and includes: Developing and testing new microbial and chemical FST methods. Determining the geographic application and animal host ranges of existing and emerging FST techniques. Conducting experimental comparisons of FST techniques. Combining direct monitoring of human pathogens associated with waterborne outbreaks and zoonotic pathogens responsible for infections among people, wildlife, or domesticated animals with the use of FST techniques. Applying FST to watershed analysis and coastal environments. Designing appropriate statistical and probability analysis of FST data and developing models for mass loadings of host-specific fecal contamination. This paper includes a critical review of FST with emphasis on the extent to which methods have been tested (especially in comparison with other methods and/or with blind samples), which methods are applicable to different situations, their shortcomings, and their usefulness in predicting public health risk or pathogen occurrence. In addition, the paper addresses the broader question of whether FST and fecal indicator monitoring is the best approach to regulate water quality and protect human health. Many FST methods have only been tested against sewage or fecal samples or isolates in laboratory studies (proof of concept testing) and/or applied in field studies where the “real” answer is not known, so their comparative performance and accuracy cannot be assessed. For FST to be

  16. Cryptosporidium, chronic diarrhoea and the proximal small intestinal mucosa.

    PubMed Central

    Phillips, A D; Thomas, A G; Walker-Smith, J A

    1992-01-01

    The association between Cryptosporidium, chronic diarrhoea and a proximal small intestinal mucosal enteropathy was reviewed over a six and a half year period. One hundred and twenty three children with cryptosporidiosis and no clinical evidence of immune deficiency were identified. 50% of children excreting only Cryptosporidium had chronic diarrhoea. Most cases (63%) of chronic diarrhoea occurred in the first two years of life. A mild to moderate enteropathy was present in all nine children undergoing a small intestinal biopsy and seven showed the presence of Cryptosporidium adhering to villous epithelium. All patients eventually recovered spontaneously. Cryptosporidium is a cause of chronic diarrhoea and a proximal small intestinal mucosal enteropathy in children without immune deficiency. Screening for the parasite should be part of the investigative procedures in children with chronic diarrhoea. Images Figure 4 PMID:1398230

  17. Giardia and Cryptosporidium join the 'Neglected Diseases Initiative'.

    PubMed

    Savioli, L; Smith, H; Thompson, A

    2006-05-01

    Giardia and Cryptosporidium are ubiquitous enteric protozoan pathogens that infect humans, domestic animals and wildlife worldwide. Both pathogens are significant causes of diarrhea and nutritional disorders in institutional and community settings. They are also significant waterborne pathogens. In developing regions of the world, Giardia and Cryptosporidium constitute part of the complex group of parasitic, bacterial and viral diseases that impair the ability to achieve full potential and impair development and socio-economic improvements. All diseases included in the WHO Neglected Diseases Initiative have a common link with poverty and, as the current view is to take a comprehensive approach to all these diseases, both Giardia and Cryptosporidium were included in 2004. Our current state of knowledge of Giardia and Cryptosporidium is summarized here, and some important questions are raised that need to be addressed if control strategies are to be effective.

  18. Clinical and subclinical infections with Giardia and Cryptosporidium in animals

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Giardia and Cryptosporidium are frequent parasites of livestock, companion animals, and wildlife, raising questions about the clinical significance of such infections. Infections with both parasites have a wide spectrum of symptoms that can vary between asymptomatic infections to serious infection ...

  19. Molecular identification of Giardia and Cryptosporidium from dogs and cats.

    PubMed

    Sotiriadou, Isaia; Pantchev, Nikola; Gassmann, Doreen; Karanis, Panagiotis

    2013-01-01

    The aim of the present study was to diagnose the presence of Giardia cysts and Cryptosporidium oocysts in household animals using nested polymerase chain reaction (PCR) and sequence analysis. One hundred faecal samples obtained from 81 dogs and 19 cats were investigated. The Cryptosporidium genotypes were determined by sequencing a fragment of the small subunit (SSU) rRNA gene, while the Giardia Assemblages were determined through analysis of the glutamate dehydrogenase (GDH) locus. Isolates from five dogs and two cats were positive by PCR for the presence of Giardia, and their sequences matched the zoonotic Assemblage A of Giardia. Cryptosporidium spp. isolated from one dog and one cat were both found to be C. parvum. One dog isolate harboured a mixed infection of C. parvum and Giardia Assemblage A. These findings support the growing evidence that household animals are potential reservoirs of the zoonotic pathogens Giardia spp. and Cryptosporidium spp. for infections in humans.

  20. Interlaboratory comparison of three microbial source tracking quantitative polymerase chain reaction (qPCR) assays from fecal-source and environmental samples

    USGS Publications Warehouse

    Stelzer, Erin A.; Strickler, Kriston M.; Schill, William B.

    2012-01-01

    During summer and early fall 2010, 15 river samples and 6 fecal-source samples were collected in West Virginia. These samples were analyzed by three laboratories for three microbial source tracking (MST) markers: AllBac, a general fecal indicator; BacHum, a human-associated fecal indicator; and BoBac, a ruminant-associated fecal indicator. MST markers were analyzed by means of the quantitative polymerase chain reaction (qPCR) method. The aim was to assess interlaboratory precision when the three laboratories used the same MST marker and shared deoxyribonucleic acid (DNA) extracts of the samples, but different equipment, reagents, and analyst experience levels. The term assay refers to both the markers and the procedure differences listed above. Interlaboratory precision was best for all three MST assays when using the geometric mean absolute relative percent difference (ARPD) and Friedman's statistical test as a measure of interlaboratory precision. Adjustment factors (one for each MST assay) were calculated using results from fecal-source samples analyzed by all three laboratories and applied retrospectively to sample concentrations to account for differences in qPCR results among labs using different standards and procedures. Following the application of adjustment factors to qPCR results, ARPDs were lower; however, statistically significant differences between labs were still observed for the BacHum and BoBac assays. This was a small study and two of the MST assays had 52 percent of samples with concentrations at or below the limit of accurate quantification; hence, more testing could be done to determine if the adjustment factors would work better if the majority of sample concentrations were above the quantification limit.

  1. Tracking the source of mercury in coastal populations of California Cougars (puma concolor)

    NASA Astrophysics Data System (ADS)

    Weiss-Penzias, P. S.; Wilmers, C.; Yovovich, V.; Houghtaling, P.; Torregrosa, A.

    2015-12-01

    As part of a project on the cycling of mercury (Hg) from the ocean to fog and deposition to land in coastal California, the whiskers of pumas from coastal and inland populations in California were analyzed for total Hg (HgT). Previous studies have shown that fog water in coastal California contains enhanced concentrations of monomethyl Hg (MMHg) compared to rain water. The likely source of fog MMHg is from evasion and demethylation of dimethyl Hg (DMHg) from coastal ocean upwelling. The California coast receives seasonal inputs of fog drip, and we hypothesized that if fog water deposition of MMHg was making an impact, the observable effects might be seen in high trophic level predators of the terrestrial ecosystem. Puma whiskers from 88 individuals from the Santa Cruz Mountains, a sub-range of the California Coast Range, were obtained and compared with puma whiskers from 12 individuals from the foothills of the Sierra Nevada Mountains. Mean total Hg in puma whiskers from the coastal population is 1.0 ± 1.5 ug Hg / g whisker (ppm), whereas mean HgT from the inland puma population is 0.13 ± 0.09 ppm. The difference between these means is significant to the 95% confidence level. For the coastal puma population, the whiskers from 10 individuals had HgT concentrations > 2.0 ppm and 3 individuals had HgT > 4 ppm, which exceeds the U.S. EPA reference dose for humans (1 ppm) approaches a level of concern found for other large mammals such as polar bears (5 ppm). The study is ongoing and HgT concentrations will be determined in the fur and flesh of deer from the same locations as the puma whiskers, since deer comprise ~95% of the puma diet. Samples of plants that are likely fed upon by deer that span the coastal-inland transect will also be analyzed for HgT. Estimates of fog frequency spatial patterns, derived from weather satellite observations and topographic modeling, will be compared with the HgT content of plant and animal tissue in coastal California to quantify

  2. Cryptosporidium ryanae n. sp. (Apicomplexa: Cryptosporidiidae) in cattle (Bos taurus).

    PubMed

    Fayer, Ronald; Santín, Mónica; Trout, James M

    2008-10-01

    A new species, Cryptosporidium ryanae, is described from cattle. Oocysts of C. ryanae, previously identified as the Cryptosporidium deer-like genotype and recorded as such in GenBank (AY587166, EU203216, DQ182597, AY741309, and DQ871345), are similar to those of Cryptosporidium parvum and Cryptosporidium bovis but smaller. This genotype has been reported to be prevalent in cattle worldwide. Oocysts obtained from a calf for the present study are the smallest Cryptosporidium oocysts reported in mammals, measuring 2.94-4.41micromx2.94-3.68microm (mean=3.16micromx3.73microm) with a length/width shape index of 1.18 (n=40). The pre-patent period for two Cryptosporidium-naïve calves fed C. ryanae oocysts was 11 days and the patent period was 15-17 days. Oocysts were not infectious for BALB/c mice or lambs. Fragments of the SSU-rDNA, HSP-70, and actin genes amplified by PCR were purified and PCR products were sequenced. Multi-locus analysis of the three unlinked loci demonstrated the new species to be distinct from all other species and also demonstrated a lack of recombination, providing further evidence of species status. Based on morphological, molecular and biological data, this geographically widespread parasite found only in Bos taurus calves is recognized as a new species and is named C. ryanae.

  3. Cryptosporidium erinacei n. sp. (Apicomplexa: Cryptosporidiidae) in hedgehogs.

    PubMed

    Kváč, Martin; Hofmannová, Lada; Hlásková, Lenka; Květoňová, Dana; Vítovec, Jiří; McEvoy, John; Sak, Bohumil

    2014-03-17

    The morphological, biological, and molecular characteristics of Cryptosporidium hedgehog genotype are described, and the species name Cryptosporidium erinacei n. sp. is proposed to reflect its specificity for hedgehogs under natural and experimental conditions. Oocysts of C. erinacei are morphologically indistinguishable from Cryptosporidium parvum, measuring 4.5-5.8 μm (mean=4.9 μm) × 4.0-4.8 μm (mean=4.4 μm) with a length to width ratio of 1.13 (1.02-1.35) (n=100). Oocysts of C. erinacei obtained from a naturally infected European hedgehog (Erinaceus europaeus) were infectious for naïve 8-week-old four-toed hedgehogs (Atelerix albiventris); the prepatent period was 4-5 days post infection (DPI) and the patent period was longer than 20 days. C. erinacei was not infectious for 8-week-old SCID and BALB/c mice (Mus musculus), Mongolian gerbils (Meriones unguiculatus), or golden hamsters (Mesocricetus auratus). Phylogenetic analyses based on small subunit rRNA, 60 kDa glycoprotein, actin, Cryptosporidium oocyst wall protein, thrombospondin-related adhesive protein of Cryptosporidium-1, and heat shock protein 70 gene sequences revealed that C. erinacei is genetically distinct from previously described Cryptosporidium species.

  4. USE OF COMPOSITE DATA SETS FOR SOURCE-TRACKING ENTEROCCOCCI IN THE WATER COLUMN AND SHORELINE INTERSTITIAL WATERS ON PENSACOLA BEACH, FL

    EPA Science Inventory

    Genthner, Fred J., Joseph B. James, Diane F. Yates and Stephanie D. Friedman. Submitted. Use of Composite Data Sets for Source-Tracking Enterococci in the Water Column and Shoreline Interstitial Waters on Pensacola Beach Florida. Mar. Pollut. Bull. 33 p. (ERL,GB 1212).

    So...

  5. Performance of two quantitative PCR methods for microbial source tracking of human sewage and implications for microbial risk assessment in recreational waters

    EPA Science Inventory

    Before new, rapid quantitative PCR (qPCR) methods for recreational water quality assessment and microbial source tracking (MST) can be useful in a regulatory context, an understanding of the ability of the method to detect a DNA target (marker) when the contaminant soure has been...

  6. Ultra-Stable Beacon Source for Laboratory Testing of Optical Tracking

    NASA Technical Reports Server (NTRS)

    Aso, Yoichi; Marka, Szabolcs; Kovalik, Joseph

    2008-01-01

    The ultra-stable beacon source (USBS) provides a laser-beam output with a very low angular jitter and can be used as an absolute angular reference to simulate a beacon in the laboratory. The laser is mounted on the top of a very short (approximately equal to 1 m) inverted pendulum (IP) with its optical axis parallel to the carbon fiber pendulum leg. The 85-cm, carbon fiber rods making up the leg are very lightweight and rigid, and are supported by a flex-joint at the bottom (see figure). The gimbal-mounted laser is a weight-adjustable load of about 1.5 kg with its center of rotation co-located with the center of percussion of the inverted pendulum. This reduces the coupling of transverse motion at the base of the pendulum to angular motion of the laser at the top. The inverted pendulum is mounted on a gimbal with its center of rotation coinciding with the pivot position of the inverted pendulum flexure joint. This reduces coupling of ground tilt at the inverted pendulum base to motion of the laser mounted at the top. The mass of the top gimbal is adjusted to give the pendulum a very low resonant frequency (approximately equal to 10 mHz) that filters transverse seismic disturbances from the ground where the base is attached. The motion of the IP is monitored by an optical-lever sensor. The laser light is reflected by the mirror on the IP, and then is detected by a quadrant photo-detector (QPD). The position of the beam spot on the QPD corresponds to the tilt of the IP. Damping of this motion is provided by two coil and magnet pairs. The bottom gimbal mount consists of two plates. The IP is mounted on the second plate. The first plate is supported by two posts through needles and can be rotated about the axis connecting the tips of the needles. The second plate hangs from the first plate and can be rotated about the axis perpendicular to the first plate. As a result, the second plate acts as a two-axis rotation stage. Its center of rotation is located at the

  7. Pathogenic bacteria and microbial-source tracking markers in Brandywine Creek Basin, Pennsylvania and Delaware, 2009-10

    USGS Publications Warehouse

    Duris, Joseph W.; Reif, Andrew G.; Olson, Leif E.; Johnson, Heather E.

    2011-01-01

    virulent type of pathogenic E. coli, was found only in the West Branch and main stem at high flow but was not found in the East Branch under similar conditions. However, it must be noted that throughout the entire year of sampling there were occasions, during both high and normal flows, when both the East and West Branches were potential contributors of pathogen and microbial-source tracking markers to the main stem. Therefore, this study indicates that under selected conditions (high flow, October through March), West Branch Brandywine Creek Basin was the most likely source of elevated FIB densities in the main stem. These elevated densities are associated with more frequent detection of selected pathogenic E. coli markers (rfbO157 stx1) and are associated with MST markers of bovine source. However, during other times of the year, both the West Branch and East Branch Basins are acting as potential sources of FIB and fecally derived pathogens.

  8. Detection and characterisation of Giardia and Cryptosporidium in Hungarian raw, surface and sewage water samples by IFT, PCR and sequence analysis of the SSUrRNA and GDH genes.

    PubMed

    Plutzer, Judit; Karanis, Panagiotis; Domokos, Klarissza; Törökné, Andrea; Márialigeti, Károly

    2008-10-01

    We investigated the prevalence of Giardia and Cryptosporidium species and analysed the genotypes in 36 samples collected from different water sources and various geographic areas in Hungary. Samples were collected from drinking water and sewage treatment plants and from the recreation area of Lake Balaton. The (oo)cysts were purified according to the US EPA 1623 method and they were detected by immunofluorescence test (IFT). Genomic DNA was extracted from all samples and then the GDH target gene for Giardia and the SSUrDNA for both Giardia and for Cryptosporidium species were amplified by PCR. 24 out of 36 samples (67%) were Giardia positive and 15 (42%) were Cryptosporidium positive by IFT. PCR confirmed that 13 out of 36 samples (36%) were Giardia positive and 10 (28%) contained Cryptosporidium. Twelve Giardia and two Cryptosporidium PCR products were successfully sequenced. In seven samples G. lamblia Assemblage A and in one sample Assemblage B and in four cases Assemblages A and B have been found. In one sample C. parvum and in the other separate sample C. meleagridis were detected. Sequence analysis revealed a new subtype of G. duodenalis complex, clustered close to the Assemblage A group. This study provides the first report on simultaneous detection and genotyping of G. duodenalis and Cryptosporidium species from water supplies in Hungary.

  9. Tracking and Interception of Ground-Based RF Sources Using Autonomous Guided Munitions with Passive Bearings-Only Sensors and Tracking Algorithms

    DTIC Science & Technology

    2006-01-01

    manipulation of the measurement equation and has very good convergence properties for stationary targets (Aidala, Nardone ) [7]. However, the PLLS does lead to...Applications,” IEEE Transactions on Aerospace and Electronic Systems, vol. 15, no. 1, January 1979. 4. Nardone , S. C. and Aidala, V. J., “Observability...July 1978. 8. Aidala, V. J. and Nardone , S. C., “Biased Estimation Properties of the Pseudolinear Tracking Filter,” IEEE Transactions on Aerospace

  10. 40 CFR 141.713 - Schedule for compliance with Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Cryptosporidium treatment requirements. 141.713 Section 141.713 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Treatment Technique Requirements § 141.713 Schedule for compliance...

  11. Microscopic and Molecular Detection of Cryptosporidium andersoni and Cryptosporidium xiaoi in Wastewater Samples of Tehran Province, Iran

    PubMed Central

    HATAM-NAHAVANDI, Kareem; MOHEBALI, Mehdi; MAHVI, Amir-Hossein; KESHAVARZ, Hossein; NAJAFIAN, Hamid-Reza; MIRJALALI, Hamed; REZAEI, Sasan; REZAEIAN, Mostafa

    2016-01-01

    Background: As a waterborne pathogen, Cryptosporidium is one of the most common causes of gastroenteritis in human and hoofed livestock animals. This study aimed to investigate the distribution of Cryptosporidium spp. in human and livestock wastewaters in Iran, by the 18S rRNA sequence analysis. Methods: A total of 54 raw wastewater samples collected from three urban treatment plants and two slaughterhouses during 2014–2015 in Tehran, Iran. The presence of the Cryptosporidium oocysts was assessed by immunofluorescence with monoclonal antibodies. To characterize the oocysts at the molecular level, the 18S rRNA gene of Cryptosporidium was PCR amplified and sequenced. Results: Of the 54 wastewater samples examined, 34 (62.9%) were positive for Cryptosporidium oocysts using the IFA. Of these, 70.5% (24/34) were positive by PCR, that 91.6% (22/24) were successfully sequenced. The species of C. andersoni (95.4%) and C. xiaoi (4.6%) were detected in livestock wastewater samples. Conclusion: C. andersoni was the major Cryptosporidium sp. found in the aquatic environmental wastewater samples. The high rate of detection of C. andersoni in domestic wastewater was probably the result of the predominancy of this species in cattle herds in Iran. The current study is the first report of C. xiaoi in Iran. PMID:28127361

  12. Prospects for immunotherapy and vaccines against Cryptosporidium.

    PubMed

    Mead, Jan R

    2014-01-01

    Cryptosporidium spp is a ubiquitous parasite that has long been recognized as a frequent cause of protozoal diarrhea in humans. While infections in immunocompetent hosts are usually self-limiting, immunocompromised individuals can develop severe, chronic, and life-threatening illness. Vaccine development or immunotherapy that prevents disease or reduces the severity of infection is a relevant option since efficacious drug treatments are lacking. In particular, children in developing countries might benefit the most from a vaccine since cryptosporidiosis in early childhood has been reported to be associated with subsequent impairment in growth, physical fitness, and intellectual capacity. In this review, immunotherapies that have been used clinically are described as well as experimental vaccines and their evaluation in vivo.

  13. Evaluation of two spike-and-recovery controls for assessment of extraction efficiency in microbial source tracking studies

    USGS Publications Warehouse

    Stoeckel, D.M.; Stelzer, E.A.; Dick, L.K.

    2009-01-01

    Quantitative PCR (qPCR), applied to complex environmental samples such as water, wastewater, and feces, is susceptible to methodological and sample related biases. In this study, we evaluated two exogenous DNA spike-and-recovery controls as proxies for recovery efficiency of Bacteroidales 16S rDNA gene sequences (AllBac and qHF183) that are used for microbial source tracking (MST) in river water. Two controls-(1) the plant pathogen Pantoea stewartii, carrying the chromosomal target gene cpsD, and (2) Escherichia coli, carrying the plasmid-borne target gene DsRed2-were added to raw water samples immediately prior to concentration and DNA extraction for qPCR. When applied to samples processed in replicate, recovery of each control was positively correlated with the observed concentration of each MST marker. Adjustment of MST marker concentrations according to recovery efficiency reduced variability in replicate analyses when consistent processing and extraction methodologies were applied. Although the effects of this procedure on accuracy could not be tested due to uncertainties in control DNA concentrations, the observed reduction in variability should improve the strength of statistical comparisons. These findings suggest that either of the tested spike-and-recovery controls can be useful to measure efficiency of extraction and recovery in routine laboratory processing. ?? 2009 Elsevier Ltd.

  14. USE OF LONG ACTING STEROID METHYLPREDNISOLONE ACETATE FOR THE PROLONGATION OF CRYPTOSPORIDIUM SP. IN MICE

    EPA Science Inventory

    Current protocols for Cryptosporidium sp. propagation in mice vary according to the strain and age of mouse as well as the species of Cryptosporidium. Cryptosporidium muris is a natural parasite of mice, but only neonatal mice are susceptible to C. parvum infection. Published C...

  15. A new genotype of Cryptosporidium from giant panda (Ailuropoda melanoleuca) in China.

    PubMed

    Liu, Xuehan; He, Tingmei; Zhong, Zhijun; Zhang, Hemin; Wang, Rongjun; Dong, Haiju; Wang, Chengdong; Li, Desheng; Deng, Jiabo; Peng, Guangneng; Zhang, Longxian

    2013-10-01

    Fifty-seven fecal samples were collected from giant pandas (Ailuropoda melanoleuca) in the China Conservation and Research Centre for the Giant Panda (CCRCGP) in Sichuan and examined for Cryptosporidium oocysts by Sheather's sugar flotation technique. An 18-year-old male giant panda was Cryptosporidium positive, with oocysts of an average size of 4.60×3.99 μm (n=50). The isolate was genetically analyzed using the partial 18S rRNA, 70 kDa heat shock protein (HSP70), Cryptosporidium oocyst wall protein (COWP) and actin genes. Multi-locus genetic characterization indicated that the present isolate was different from known Cryptosporidium species and genotypes. The closest relative was the Cryptosporidium bear genotype, with 11, 10, and 6 nucleotide differences in the 18S rRNA, HSP70, and actin genes, respectively. Significant differences were also observed in the COWP gene compared to Cryptosporidium mongoose genotype. The homology to the bear genotype at the 18S rRNA locus was 98.6%, which is comparable to that between Cryptosporidium parvum and Cryptosporidium hominis (99.2%), or between Cryptosporidium muris and Cryptosporidium andersoni (99.4%). Therefore, the Cryptosporidium in giant pandas in this study is considered as a new genotype: the Cryptosporidium giant panda genotype.

  16. MICROBIAL SOURCE TRACKING GUIDE

    EPA Science Inventory

    Fecal contamination of surface waters used for recreation, drinking water and aquaculture are a continuous environmental problem and pose significant human health risks. An alarming amount of the United States rivers/streams (39%), lakes (45%), and estuaries (51%) are not safe f...

  17. BACTERIAL SOURCE TRACKING

    EPA Science Inventory

    Fecal contamination of surface waters used for recreation, drinking water and aquaculture are a continuous environmental problem and pose significant human health risks. An alarming amount of the United States rivers/streams (39%), lakes (45%), and estuaries (51%) are not safe f...

  18. BACTERIAL SOURCE TRACKING

    EPA Science Inventory

    Fecal contamination of surface waters used for recreation, drinking water and aquaculture are a continuous environmental problem and pose significant human health risks. An alarming amount of the US rivers/streams (39%) lakes (45%), and estuaries (51%) are not safe for fishing an...

  19. MICROBIAL SOURCE TRACKING - 101

    EPA Science Inventory

    Fecal contamination of surface waters used for recreation, drinking water and aquaculture are a continuous environmental problem and pose significant human health risks. Today, a large portion of the United States rivers/streams (39%), lakes (45%), and estuaries (51%) are not saf...

  20. MICROBIAL SOURCE TRACKING - 2005

    EPA Science Inventory

    Fecal contamination of surface waters used for recreation, drinking water and aquaculture are a continuous environmental problem and pose significant human health risks. An alarming amount of the United States rivers/streams (39%), lakes (45%), and estuaries (51%) are not safe f...

  1. Microbial Source Tracking

    EPA Science Inventory

    Bacterial indicators of fecal contamination provide the basis for assessing the microbial quality of environmental waters. While the indicator concept has overall helped reduce waterborne outbreaks in recreational waters, the public health value of currently used indicator bacter...

  2. Occurrence of Cryptosporidium and Giardia in recycled waters used for irrigation and first description of Cryptosporidium parvum and C. muris in Greece.

    PubMed

    Spanakos, Gregory; Biba, Anastasia; Mavridou, Athena; Karanis, Panagiotis

    2015-05-01

    Here, we present the first time findings regarding the occurrence of Cryptosporidium and Giardia in sewage waters and the first molecular characterization of Cryptosporidium species in Greece. Biological treatment plants from three regions in Greece have been investigated. The detection of Cryptosporidium oocysts was by modified Ziehl-Neelsen acid fast (MZN-AF) and by immunofluorescence microscopy (IFT) for Cryptosporidium and Giardia (oo)cysts, whereas nested PCR based on the SSU rDNA assay was used for molecular detection of Cryptosporidium followed by sequencing for the genetic characterization of the species. In total, 73 samples (37 raw sewage samples and 38 of treated water samples) were collected and analyzed. Of the 73 water samples, 4 samples were Cryptosporidium-positive by IFT and staining, 12 samples were Cryptosporidium-positive by nested PCR; 9 samples were Giardia-positive by IFT. We showed that Cryptosporidium cysts are found both in the input and the discharge of the biological treatment plants. Molecular characterization of Cryptosporidium based on the small subunit ribosomal DNA gene resulted in the determination of Cryptosporidium parvum and Cryptosporidium muris Greek isolates. This is the first report of Cryptosporidium and Giardia occurrence in wastewaters and the first molecular identification of Cryptosporidium species in Greek environments. As the treated water is used for irrigation, or it is discharged into the sea, our findings indicate that biological treatment facilities constitute a possible risk for public health because the related species are prevalent in humans; the results invite for further epidemiological investigations to evaluate the real public health risk in Greece.

  3. Cryptosporidium hominis gene catalog: a resource for the selection of novel Cryptosporidium vaccine candidates

    PubMed Central

    Ifeonu, Olukemi O.; Simon, Raphael; Tennant, Sharon M.; Sheoran, Abhineet S.; Daly, Maria C.; Felix, Victor; Kissinger, Jessica C.; Widmer, Giovanni; Levine, Myron M.; Tzipori, Saul; Silva, Joana C.

    2016-01-01

    Human cryptosporidiosis, caused primarily by Cryptosporidium hominis and a subset of Cryptosporidium parvum, is a major cause of moderate-to-severe diarrhea in children under 5 years of age in developing countries and can lead to nutritional stunting and death. Cryptosporidiosis is particularly severe and potentially lethal in immunocompromised hosts. Biological and technical challenges have impeded traditional vaccinology approaches to identify novel targets for the development of vaccines against C. hominis, the predominant species associated with human disease. We deemed that the existence of genomic resources for multiple species in the genus, including a much-improved genome assembly and annotation for C. hominis, makes a reverse vaccinology approach feasible. To this end, we sought to generate a searchable online resource, termed C. hominis gene catalog, which registers all C. hominis genes and their properties relevant for the identification and prioritization of candidate vaccine antigens, including physical attributes, properties related to antigenic potential and expression data. Using bioinformatic approaches, we identified ∼400 C. hominis genes containing properties typical of surface-exposed antigens, such as predicted glycosylphosphatidylinositol (GPI)-anchor motifs, multiple transmembrane motifs and/or signal peptides targeting the encoded protein to the secretory pathway. This set can be narrowed further, e.g. by focusing on potential GPI-anchored proteins lacking homologs in the human genome, but with homologs in the other Cryptosporidium species for which genomic data are available, and with low amino acid polymorphism. Additional selection criteria related to recombinant expression and purification include minimizing predicted post-translation modifications and potential disulfide bonds. Forty proteins satisfying these criteria were selected from 3745 proteins in the updated C. hominis annotation. The immunogenic potential of a few of these is

  4. Decay of Fecal Indicator Bacterial Populations and Bovine-Associated Source-Tracking Markers in Freshly Deposited Cow Pats

    PubMed Central

    Oladeinde, Adelumola; Bohrmann, Thomas; Wong, Kelvin; Purucker, S. T.; Bradshaw, Ken; Brown, Reid; Snyder, Blake

    2014-01-01

    Understanding the survival of fecal indicator bacteria (FIB) and microbial source-tracking (MST) markers is critical to developing pathogen fate and transport models. Although pathogen survival in water microcosms and manure-amended soils is well documented, little is known about their survival in intact cow pats deposited on pastures. We conducted a study to determine decay rates of fecal indicator bacteria (Escherichia coli and enterococci) and bovine-associated MST markers (CowM3, Rum-2-bac, and GenBac) in 18 freshly deposited cattle feces from three farms in northern Georgia. Samples were randomly assigned to shaded or unshaded treatment in order to determine the effects of sunlight, moisture, and temperature on decay rates. A general linear model (GLM) framework was used to determine decay rates. Shading significantly decreased the decay rate of the E. coli population (P < 0.0001), with a rate of −0.176 day−1 for the shaded treatment and −0.297 day−1 for the unshaded treatment. Shading had no significant effect on decay rates of enterococci, CowM3, Rum-2-bac, and GenBac (P > 0.05). In addition, E. coli populations showed a significant growth rate (0.881 day−1) in the unshaded samples during the first 5 days after deposition. UV-B was the most important parameter explaining the decay rate of E. coli populations. A comparison of the decay behaviors among all markers indicated that enterococcus concentrations exhibit a better correlation with the MST markers than E. coli concentrations. Our results indicate that bovine-associated MST markers can survive in cow pats for at least 1 month after excretion, and although their decay dynamic differs from the decay dynamic of E. coli populations, they seem to be reliable markers to use in combination with enterococci to monitor fecal pollution from pasture lands. PMID:24141130

  5. Childhood Cryptosporidium infection among aboriginal communities in Peninsular Malaysia

    PubMed Central

    Al-Mekhlafi, H M; Mahdy, M A K; ’Azlin, M Y; Fatmah, M S; Norhayati, M

    2011-01-01

    Cryptosporidium is a coccidian parasite that is prevalent worldwide, some species of which cause morbidity in both immunocompromised and immunocompetent individuals. The prevalence and predictors of Cryptosporidium infection, and its effect on nutritional status, have recently been explored among 276 children (141 boys and 135 girls, aged 2–15 years) in aboriginal (Orang Asli) villages in the Malaysian state of Selangor. Faecal smears were examined by the modified Ziehl–Neelsen staining technique while socio–economic data were collected using a standardized questionnaire. Nutritional status was assessed by anthropometric measurements. Cryptosporidium infection, which was detected in 7·2% of the aboriginal children, was found to be significantly associated with low birthweight (⩽2·5 kg), being part of a large household (with more than seven members) and prolonged breast feeding (>2 years). The output of a binary logistic regression confirmed that large household size was a significant predictor of Cryptosporidium infection (giving an odds ratio of 2·15, with a 95% confidence interval of 1·25–5·02). Cryptosporidium infection is clearly a public-health problem among the aboriginal children of Selangor, with person-to-person the most likely mode of transmission. PMID:21396250

  6. Giardia and Cryptosporidium in cetaceans on the European Atlantic coast.

    PubMed

    Reboredo-Fernández, Aurora; Ares-Mazás, Elvira; Martínez-Cedeira, José A; Romero-Suances, Rafael; Cacciò, Simone M; Gómez-Couso, Hipólito

    2015-02-01

    The occurrence of Giardia and Cryptosporidium was investigated in cetacean specimens stranded on the northwestern coast of Spain (European Atlantic coast) by analysis of 65 samples of large intestine from eight species. The parasites were identified by direct immunofluorescence antibody test (IFAT) and by PCR amplification of the β-giardin gene, the ITS1-5.8S-ITS2 region and the SSU-rDNA gene of Giardia and the SSU-rDNA gene of Cryptosporidium. Giardia and Cryptosporidium were detected in 7 (10.8 %) and 9 samples (13.8 %), respectively. In two samples, co-infection with both parasites was observed. Giardia duodenalis assemblages A, C, D and F, and Cryptosporidium parvum were identified. This is the first report of G. duodenalis in Balaenoptera acutorostrata, Kogia breviceps and Stenella coeruleoalba and also the first report of Cryptosporidium sp. in B. acutorostrata and of C. parvum in S. coeruleoalba and Tursiops truncatus. These results extend the known host range of these waterborne enteroparasites.

  7. Epidemiology of Cryptosporidium infection in cattle in China: a review

    PubMed Central

    Gong, Chao; Cao, Xue-Feng; Deng, Lei; Li, Wei; Huang, Xiang-Ming; Lan, Jing-Chao; Xiao, Qi-Cheng; Zhong, Zhi-Jun; Feng, Fan; Zhang, Yue; Wang, Wen-Bo; Guo, Ping; Wu, Kong-Ju; Peng, Guang-Neng

    2017-01-01

    The present review discusses the findings of cryptosporidiosis research conducted in cattle in China and highlights the currently available information on Cryptosporidium epidemiology, genetic diversity, and distribution in China, which is critical to understanding the economic and public health importance of cryptosporidiosis transmission in cattle. To date, 10 Cryptosporidium species have been detected in cattle in China, with an overall infection rate of 11.9%. The highest rate of infection (19.5%) was observed in preweaned calves, followed by that in juveniles (10.69%), postweaned juveniles (9.0%), and adult cattle (4.94%). The dominant species were C. parvum in preweaned calves and C. andersoni in postweaned, juvenile, and adult cattle. Zoonotic Cryptosporidium species (C. parvum and C. hominis) were found in cattle, indicating the possibility of transmission between humans and cattle. Different cattle breeds had significant differences in the prevalence rate and species of Cryptosporidium. This review demonstrates an age-associated, breed-associated, and geographic-related occurrence of Cryptosporidium and provides references for further understanding of the epidemiological characteristics, and for preventing and controlling the disease. PMID:28098070

  8. Characterization of a gamma-ray tracking array: A comparison of GRETINA and Gammasphere using a 60Co source

    NASA Astrophysics Data System (ADS)

    Lauritsen, T.; Korichi, A.; Zhu, S.; Wilson, A. N.; Weisshaar, D.; Dudouet, J.; Ayangeakaa, A. D.; Carpenter, M. P.; Campbell, C. M.; Clément, E.; Crawford, H. L.; Cromaz, M.; Fallon, P.; Greene, J. P.; Janssens, R. V. F.; Khoo, T. L.; Lalović, N.; Lee, I. Y.; Macchiavelli, A. O.; Perez-Vidal, R. M.; Pietri, S.; Radford, D. C.; Ralet, D.; Riley, L. A.; Seweryniak, D.; Stezowski, O.

    2016-11-01

    In this paper, we provide a formalism for the characterization of tracking arrays with emphasis on the proper corrections required to extract their photopeak efficiencies and peak-to-total ratios. The methods are first applied to Gammasphere, a well characterized 4π array based on the principle of Compton suppression, and subsequently to GRETINA. The tracking efficiencies are then discussed and some guidelines as to what clustering angle to use in the tracking algorithm are presented. It was possible, using GEANT4 simulations, to scale the measured efficiencies up to the expected values for the full 4π implementation of GRETA.

  9. Chasing the Black Smoke: Building Software for CALIPSO Satellite Data to Aid in Tracking and Identifying Sources of Aerosols and their Impact on the Earth's Climate

    NASA Astrophysics Data System (ADS)

    Mercer, G. A.

    2015-12-01

    The Cloud-Aerosol Lidar and Infrared Pathfinder Satellite Observation (CALIPSO) satellite is a NASA Earth observation that analyzes aerosol particles suspended in the Earth's atmosphere. Researchers use visualized CALIPSO data to track the global distribution, dispersion, and source of aerosols. There currently exists a tool for displaying CALIPSO data, but this tool does not support needed features for tracking aerosols such as selecting regions of data and sharing those selected regions, making tracking specific airborne objects difficult for researchers. Adding these necessary features to the current CALIPSO visualization tool is difficult, as the tool is written in Interactive Data Language (IDL), a proprietary and obscure language and writing additional features for the tool would require a specialized development team. This topic will focus on release of a new tool for visualization CALIPSO's atmospheric data, or the Visualization of CALIPSO (VOCAL) open source Python program. The talk will explain why VOCAL will serve as the successor to the current visualization tool for CALIPSO data, what new features VOCAL brings to the table for researchers, and how this new tool can further support the tracking and identification of aerosols in the Earth's atmosphere.

  10. Identifying human and livestock sources of fecal contamination in Kenya with host-specific Bacteroidales assays.

    PubMed

    Jenkins, Marion W; Tiwari, Sangam; Lorente, Mario; Gichaba, Charles Maina; Wuertz, Stefan

    2009-11-01

    Microbial source tracking to distinguish between human, livestock and wildlife fecal pollution using molecular techniques is a rapidly evolving approach in many developed countries, but has not previously been applied on the African continent. DNA extracts from cow, donkey, and human fecal specimens and raw domestic sewage samples collected in Kenya were tested against five existing quantitative PCR assays designed to detect universal (2), human-specific (2), and cow-specific (1) fecal Bacteroidales genetic markers. Water samples from the River Njoro in Kenya were evaluated using the five tested Bacteroidales markers and a multi-species assay for Cryptosporidium in a preliminary exploration of fecal pollution sources and health risks in this watershed. Diagnostic sensitivity on the validation set varied from 18 to 100% for the five assays while diagnostic specificity was 100%. Of the 2 universal assays, Total Bacteroidales [Dick, L.K, Field, K.G., 2004. Rapid estimation of numbers of fecal Bacteroidetes by use of a quantitative PCR assay for 16S rRNA genes. Appl. Environ. Microbiol. 70, 5695-5697] showed lower generic fecal diagnostic sensitivity, at 55%, than BacUni-UCD, at 100%, in detecting fecal markers on the 42-sample validation set. Human-specific assay HF183 demonstrated 65% sensitivity overall, and 80% on the human sewage samples, compared to 18% overall and 0% sewage for human-specific assay BacHum-UCD. Cow-specific assay BacCow-UCD had 94% sensitivity. Testing of 18 water samples indicates cows are a likely predominant source of fecal contamination in the Njoro Watershed (78% prevailing rate). Probabilistic assessment of human assay results indicates at most three of the river water samples contained human Bacteroidales. Cryptosporidium spp. markers were detected in samples from nine of the 12 sampling locations. Evidence suggesting widespread contamination by cow feces and Cryptosporidium in the Njoro watershed raises serious concerns for human and

  11. HuBac and nifH source tracking markers display a relationship to land use but not rainfall.

    PubMed

    Gentry-Shields, Jennifer; Rowny, Jakob G; Stewart, Jill R

    2012-11-15

    Identification of the source of fecal pollution is becoming a priority for states and territories in the U.S. in order to meet water quality standards and to develop and implement total maximum daily loads. The goal of this research was to relate microbial source tracking (MST) assay concentrations to land use and levels of impervious surfaces in order to gauge how increasing development is associated with human fecal contamination in inland watersheds. The concentrations of two proposed MST markers, targeting nifH of Methanobrevibacter smithii and HuBac of Bacteroides sp., were positively correlated with increasing anthropogenic development and impervious surfaces. Higher concentrations of these MST markers in more urbanized watersheds suggest that increasing development negatively affects water quality. Neither MST marker concentration was correlated with antecedent rainfall levels, and detection of markers did not differ between dry weather and rain events. Water samples were also analyzed for norovirus and enterovirus, but these enteric viruses were rarely detected. These MST results differ from previous studies that have found correlations between traditional fecal indicator bacteria (FIB) and antecedent rainfall. This difference suggests that the MST markers used in this study may be more specific for recent, land-based contamination events as opposed to resuspension of particle-associated organisms in waterways. HuBac was detected in 98% of samples, correlating with fecal coliform and Escherichia coli concentrations. The ubiquity of the HuBac marker in our samples suggests that this marker does not provide sufficiently different or additional information than FIB, and it is likely this marker was amplifying non-human targets. The nifH marker was detected in 30% of samples. Less than half of the nifH-positive samples contained levels of fecal coliforms or E. coli above regulatory thresholds, suggesting that nifH would be more useful when utilized

  12. Cloaca prolapse and cystitis in green iguana (Iguana iguana) caused by a novel Cryptosporidium species.

    PubMed

    Kik, Marja J L; van Asten, Alphons J A M; Lenstra, Johannes A; Kirpensteijn, Jolle

    2011-01-10

    Cryptosporidium infection was associated with colitis and cystitis in 2 green iguanas (Iguana iguana). The disease was characterized by a chronic clinical course of cloacal prolapses and cystitis. Histological examination of the gut and urinary bladder showed numerous Cryptosporidium developmental stages on the surface of the epithelium with mixed inflammatory response in the lamina propria. Cryptosporidium oocysts were visualised in a cytological preparation of the faeces. Based on the small subunit ribosomal RNA gene the cryptosporidia were characterized as belonging to the intestinal cryptosporidial lineage, but not to Cryptosporidium saurophilum or Cryptosporidium serpentis species.

  13. Effect of climate change on runoff of Campylobacter and Cryptosporidium from land to surface water.

    PubMed

    Sterk, Ankie; Schijven, Jack; de Roda Husman, Ana Maria; de Nijs, Ton

    2016-05-15

    Faeces originating from wildlife, domestic animals or manure-fertilized fields, is considered an important source of zoonotic pathogens to which people may be exposed by, for instance, bathing or drinking-water consumption. An increase in runoff, and associated wash-off of animal faeces from fields, is assumed to contribute to the increase of disease outbreaks during periods of high precipitation. Climate change is expected to increase winter precipitation and extreme precipitation events during summer, but has simultaneously also other effects such as temperature rise and changes in evapotranspiration. The question is to what extent the combination of these effects influence the input of zoonotic pathogens to the surface waters. To quantitatively analyse the impacts of climate change on pathogen runoff, pathogen concentrations reaching surface waters through runoff were calculated by combining an input model for catchment pathogen loads with the Wageningen Lowland Runoff Simulator (WALRUS). Runoff of Cryptosporidium and Campylobacter was evaluated under different climate change scenarios and by applying different scenarios for sources of faecal pollution in the catchments, namely dairy cows and geese and manure fertilization. Model evaluation of these scenarios shows that climate change has little overall impact on runoff of Campylobacter and Cryptosporidium from land to the surface waters. Even though individual processes like runoff fluxes, pathogen release and dilution are affected, either positively or negatively, the net effect on the pathogen concentration in surface waters and consequently also on infection risks through recreation seems limited.

  14. Occurrence and molecular characterization of Cryptosporidium spp. in mammals and reptiles at the Lisbon Zoo.

    PubMed

    Alves, Margarida; Xiao, Lihua; Lemos, Vanessa; Zhou, Ling; Cama, Vitaliano; da Cunha, Margarida Barão; Matos, Olga; Antunes, Francisco

    2005-09-01

    The presence of Cryptosporidium parasites in mammals and reptiles kept at the Lisbon Zoo was investigated. A total of 274 stool samples were collected from 100 mammals and 29 reptiles. The species and genotype of the isolates identified by light microscopy were determined by nested PCR and sequence analysis of a fragment of the small subunit rRNA gene. Cryptosporidium oocysts were found in one black wildebeest (Connochaetes gnou), one Prairie bison (Bison bison bison) and in one Indian star tortoise (Geochelone elegans). The PCR and sequence analysis of these three isolates showed that those excreted by the Prairie bison were Cryptosporidium mouse genotype, those from the black wildebeest were from a new Cryptosporidium genotype and those infecting the Indian star tortoise were Cryptosporidium tortoise genotype. The present work reports a new Cryptosporidium genotype in a black wildebeest and the first finding of the Cryptosporidium mouse genotype in a ruminant.

  15. Molecular detection of Cryptosporidium spp. infections in water buffaloes from northeast Thailand.

    PubMed

    Inpankaew, Tawin; Jiyipong, Tawisa; Wongpanit, Kannika; Pinyopanuwat, Nongnuch; Chimnoi, Wissanuwat; Kengradomkij, Chanya; Xuan, Xuenan; Igarashi, Ikuo; Xiao, Lihua; Jittapalapong, Sathaporn

    2014-02-01

    The objectives of this study were to determine the individual and herd-level prevalence and genotype of Cryptosporidium and to identify putative risk factors associated with Cryptosporidium spp. infections in water buffaloes in northeast Thailand. Fecal samples from 600 water buffaloes of 287 farms in six provinces were collected and tested using DMSO-modified acid-fast staining and polymerase chain reaction. The overall prevalence of Cryptosporidium infections in buffaloes was 5.7 and 8.7% among individual animals and herds, respectively. The provinces with highest infected Cryptosporidium were located in the Sakon Nakhon Basin in the northern part of the region. In addition, higher herd prevalence was observed among farms with more than five buffaloes (30%) than those with five or less animals (16.2%). Thirty (88.2%) of the 34 Cryptosporidium-positive samples were Cryptosporidium parvum and four (11.8%) were Cryptosporidium ryanae.

  16. CryptoDB: a Cryptosporidium bioinformatics resource update.

    PubMed

    Heiges, Mark; Wang, Haiming; Robinson, Edward; Aurrecoechea, Cristina; Gao, Xin; Kaluskar, Nivedita; Rhodes, Philippa; Wang, Sammy; He, Cong-Zhou; Su, Yanqi; Miller, John; Kraemer, Eileen; Kissinger, Jessica C

    2006-01-01

    The database, CryptoDB (http://CryptoDB.org), is a community bioinformatics resource for the AIDS-related apicomplexan-parasite, Cryptosporidium. CryptoDB integrates whole genome sequence and annotation with expressed sequence tag and genome survey sequence data and provides supplemental bioinformatics analyses and data-mining tools. A simple, yet comprehensive web interface is available for mining and visualizing the data. CryptoDB is allied with the databases PlasmoDB and ToxoDB via ApiDB, an NIH/NIAID-fundedBioinformatics Resource Center. Recent updates to CryptoDB include the deposition of annotated genome sequences for Cryptosporidium parvum and Cryptosporidium hominis, migration to a relational database (GUS), a new query and visualization interface and the introduction of Web services.

  17. Detection of a novel genotype of Cryptosporidium in Antarctic pinnipeds.

    PubMed

    Rengifo-Herrera, Claudia; Ortega-Mora, Luis Miguel; Gómez-Bautista, Mercedes; García-Peña, Francisco Javier; García-Párraga, Daniel; Pedraza-Díaz, Susana

    2013-01-16

    A study was conducted to investigate the presence of Cryptosporidium and Giardia in Antarctic marine mammals. A total of 270 faecal samples from different species of pinnipeds from different locations in the South Shetland Islands and Antarctic Peninsula were analysed by immunofluorescence microscopy and PCR. Cryptosporidium was detected by PCR in three samples from Southern elephant seals (Mirounga leonina) and 2 Weddell seals (Leptonychotes weddellii). However, no oocysts were observed in any of the samples by immunofluorescence microscopy. Molecular characterisation of the isolates, using the 18S rDNA, the HSP70 and the COWP loci, revealed the presence of a Cryptosporidium sp., previously reported from an Antarctic Southern elephant seal, in the elephant seals and a novel genotype in Weddell seals. Giardia could not be detected in any of the samples analysed.

  18. Presence of microbial and chemical source tracking markers in roof-harvested rainwater and catchment systems for the detection of fecal contamination.

    PubMed

    Waso, M; Ndlovu, T; Dobrowsky, P H; Khan, S; Khan, W

    2016-09-01

    Microbial source tracking (MST) and chemical source tracking (CST) markers were utilized to identify fecal contamination in harvested rainwater and gutter debris samples. Throughout the sampling period, Bacteroides HF183 was detected in 57.5 % of the tank water samples and 95 % of the gutter debris samples, while adenovirus was detected in 42.5 and 52.5 % of the tank water and gutter debris samples, respectively. Human adenovirus was then detected at levels ranging from below the detection limit to 316 and 1253 genome copies/μL in the tank water and debris samples, respectively. Results for the CST markers showed that salicylic acid (average 4.62 μg/L) was the most prevalent marker (100 %) in the gutter debris samples, caffeine (average 18.0 μg/L) was the most prevalent in the tank water samples (100 %) and acetaminophen was detected sporadically throughout the study period. Bacteroides HF183 and salicylic acid (95 %) and Bacteroides HF183 and caffeine (80 %) yielded high concurrence frequencies in the gutter debris samples. In addition, the highest concurrence frequency in the tank water samples was observed for Bacteroides HF183 and caffeine (60 %). The current study thus indicates that Bacteroides HF183, salicylic acid and caffeine may potentially be applied as source tracking markers in rainwater catchment systems in order to supplement fecal indicator analyses.

  19. Evidence of Avian and Possum Fecal Contamination in Rainwater Tanks as Determined by Microbial Source Tracking Approaches

    PubMed Central

    Hamilton, K. A.; Gyawali, P.; Toze, S.; Haas, C. N.

    2016-01-01

    pathogens in rainwater tanks has been reported by several studies, supporting the necessity for the management of potential health risks. The sources of fecal pollution in rainwater tanks are unknown. However, the application of microbial source tracking (MST) markers has the potential to identify the sources of fecal contamination in a rainwater tank. In this study, we provide evidence of avian and possum fecal contamination in tank water samples using molecular markers. This study established a potential link between the degradation of the microbial quality of tank water and avian and possum feces. PMID:27208100

  20. Application of human and animal viral microbial source tracking tools in fresh and marine waters from five different geographical areas.

    PubMed

    Rusiñol, Marta; Fernandez-Cassi, Xavier; Hundesa, Ayalkibet; Vieira, Carmen; Kern, Anita; Eriksson, Irene; Ziros, Panos; Kay, David; Miagostovich, Marize; Vargha, Marta; Allard, Annika; Vantarakis, Apostolos; Wyn-Jones, Peter; Bofill-Mas, Sílvia; Girones, Rosina

    2014-08-01

    Integrated river basin management planning to mitigate the impacts of economic, demographic and climate change is an important issue for the future protection of water resources. Identifying sources of microbial contamination via the emerging science of Microbial Source Tracking (MST) plays a key role in risk assessment and the design of remediation strategies. Following an 18-month surveillance program within the EU-FP7-funded VIROCLIME project, specific MST tools were used to assess human markers such as adenoviruses (HAdV) and JC polyomaviruses (JCPyV) and porcine and bovine markers such as porcine adenoviruses (PAdV) and bovine polyomaviruses (BPyV) via quantification with real-time PCR to analyze surface water collected from five sites within different climatic zones: the Negro River (Brazil), Glafkos River (Greece), Tisza River (Hungary), Llobregat River (Spain) and Umeälven River (Sweden). The utility of the viral MST tools and the prevalence and abundance of specific human and animal viruses in the five river catchments and adjacent seawater, which is impacted by riverine contributions from the upstream catchments, were examined. In areas where no sanitation systems have been implemented, sewage can directly enter surface waters, and river water exhibited high viral loads; HAdV and JCPyV could be detected at mean concentrations of 10(5) and 10(4) Genome Copies/Liter (GC/L), respectively. In general, river water samples upstream of urban discharges presented lower human viral loads than downstream sampling sites, and those differences appeared to increase with urban populations but decrease in response to high river flow, as the elevated river water volume dilutes microbial loads. During dry seasons, river water flow decreases dramatically, and secondary effluents can represent the bulk of the riverine discharge. We also observed that ice cover that formed over the river during the winter in the studied areas in North Europe could preserve viral stability

  1. Contamination of Atlantic coast commercial shellfish with Cryptosporidium.

    PubMed

    Fayer, R; Trout, J M; Lewis, E J; Santin, M; Zhou, L; Lal, A A; Xiao, L

    2003-01-01

    Shellfish (oysters and/or clams) were obtained from 37 commercial harvesting sites in 13 Atlantic coast states from Maine to Florida and one site in New Brunswick, Canada. Gill washings from each of 25 shellfish at each site were examined by immunofluorescence microscopy (IFA) for oocysts of Cryptosporidium. Gill washings from another 25 shellfish at each site were grouped into five pools of five shellfish each. DNA from each pool was utilized for PCR and genotyping. Oocysts were found in 3.7% of 925 oysters and clams examined by IFA in shellfish from New Brunswick and 11 of 13 states. Cryptosporidium DNA was detected by PCR in 35.2% of 185 pools. Cryptosporidium parvum genotypes 1 and 2, and Cryptosporidium meleagridis,all of which have been identified in infected humans, were identified at 37.8% of the sites. Gill washings from every site were tested for the presence of infectious oocysts by biological assay in neonatal BALB/c mice but no mice were found infected, suggesting that either the oocysts were no longer infectious or infections in mice were below the level of detection. Collectively, these findings indicate that Cryptosporidium species, indicative of pollution from human and animal feces and potentially infectious for humans, were found in commercial shellfish from 64.9% of sites examined along the Atlantic coast by either microscopy or molecular testing. Previous reports link periods of high rainfall with the elevated numbers of pathogen contaminated shellfish. Because shellfish in the present study were examined during a period of exceptionally low precipitation, the data are thought to underestimate the number of Cryptosporidium contaminated shellfish likely to be found during periods of normal or above normal precipitation.

  2. Cryptosporidium and Giardia in recreational water in Belgium.

    PubMed

    Ehsan, Md Amimul; Casaert, Stijn; Levecke, Bruno; Van Rooy, Liesbet; Pelicaen, Joachim; Smis, Anne; De Backer, Joke; Vervaeke, Bart; De Smedt, Sandra; Schoonbaert, Filip; Lammens, Saskia; Warmoes, Thierry; Geurden, Thomas; Claerebout, Edwin

    2015-09-01

    The objective of this study was to investigate the presence of Cryptosporidium and Giardia in different recreational water bodies in Belgium and to estimate the infection risk associated with swimming and other recreational activities. Cryptosporidium oocysts and/or Giardia cysts were detected in three out of 37 swimming pools, seven out of 10 recreational lakes, two out of seven splash parks and four out of 16 water fountains. In the swimming pools no infection risk for Cryptosporidium could be calculated, since oocysts were only detected in filter backwash water. The risk of Giardia infection in the swimming pools varied from 1.13×10(-6) to 2.49×10(-6) per swim per person. In recreational lakes, the infection risk varied from 2.79×10(-5) to 5.74×10(-5) per swim per person for Cryptosporidium and from 7.04×10(-5) to 1.46×10(-4) for Giardia. For other outdoor water recreation activities the estimated infection risk was 5.71×10(-6) for Cryptosporidium and 1.47×10(-5) for Giardia. However, most positive samples in the recreational lakes belonged to species/genotypes that are either animal-specific or predominantly found in animals. No Cryptosporidium was found in splash parks and water fountains, but the presence of Giardia cysts suggests a risk for human infection. The infection risk of Giardia infection during a 3.5-minute visit to a splash park for children equalled 1.68×10(-4).

  3. Long-Term Transport of Cryptosporidium Parvum

    NASA Astrophysics Data System (ADS)

    Andrea, C.; Harter, T.; Hou, L.; Atwill, E. R.; Packman, A.; Woodrow-Mumford, K.; Maldonado, S.

    2005-12-01

    The protozoan pathogen Cryptosporidium parvum is a leading cause of waterborne disease. Subsurface transport and filtration in natural and artificial porous media are important components of the environmental pathway of this pathogen. It has been shown that the oocysts of C. parvum show distinct colloidal properties. We conducted a series of laboratory studies on sand columns (column length: 10 cm - 60 cm, flow rates: 0.7 m/d - 30 m/d, ionic strength: 0.01 - 100 mM, filter grain size: 0.2 - 2 mm, various solution chemistry). Breakthrough curves were measured over relatively long time-periods (hundreds to thousands of pore volumes). We show that classic colloid filtration theory is a reasonable tool for predicting the initial breakthrough, but it is inadequate to explain the significant tailing observed in the breakthrough of C. parvum oocyst through sand columns. We discuss the application of the Continuous Time Random Walk approach to account for the strong tailing that was observed in our experiments. The CTRW is generalized transport modeling framework, which includes the classic advection-dispersion equation (ADE), the fractional ADE, and the multi-rate mass transfer model as special cases. Within this conceptual framework, it is possible to distinguish between the contributions of pore-scale geometrical (physical) disorder and of pore-scale physico-chemical heterogeneities (e.g., of the filtration, sorption, desorption processes) to the transport of C. parvum oocysts.

  4. Diversity of Microsporidia, Cryptosporidium and Giardia in Mountain Gorillas (Gorilla beringei beringei) in Volcanoes National Park, Rwanda

    PubMed Central

    Sak, Bohumil; Petrželková, Klára J.; Květoňová, Dana; Mynářová, Anna; Pomajbíková, Kateřina; Modrý, David; Cranfield, Michael R.; Mudakikwa, Antoine; Kváč, Martin

    2014-01-01

    Background 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. Aims 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. Results 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. Conclusion 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. PMID:25386754

  5. Giardia duodenalis and Cryptosporidium occurrence in Australian sea lions (Neophoca cinerea) exposed to varied levels of human interaction.

    PubMed

    Delport, Tiffany C; Asher, Amy J; Beaumont, Linda J; Webster, Koa N; Harcourt, Robert G; Power, Michelle L

    2014-12-01

    Giardia and Cryptosporidium are amongst the most common protozoan parasites identified as causing enteric disease in pinnipeds. A number of Giardia assemblages and Cryptosporidium species and genotypes are common in humans and terrestrial mammals and have also been identified in marine mammals. To investigate the occurrence of these parasites in an endangered marine mammal, the Australian sea lion (Neophoca cinerea), genomic DNA was extracted from faecal samples collected from wild populations (n = 271) in Southern and Western Australia and three Australian captive populations (n = 19). These were screened using PCR targeting the 18S rRNA of Giardia and Cryptosporidium. Giardia duodenalis was detected in 28 wild sea lions and in seven captive individuals. Successful sequencing of the 18S rRNA gene assigned 27 Giardia isolates to assemblage B and one to assemblage A, both assemblages commonly found in humans. Subsequent screening at the gdh and β-giardin loci resulted in amplification of only one of the 35 18S rRNA positive samples at the β-giardin locus. Sequencing at the β-giardin locus assigned the assemblage B 18S rRNA confirmed isolate to assemblage AI. The geographic distribution of sea lion populations sampled in relation to human settlements indicated that Giardia presence in sea lions was highest in populations less than 25 km from humans. Cryptosporidium was not detected by PCR screening in either wild colonies or captive sea lion populations. These data suggest that the presence of G. duodenalis in the endangered Australian sea lion is likely the result of dispersal from human sources. Multilocus molecular analyses are essential for the determination of G. duodenalis assemblages and subsequent inferences on transmission routes to endangered marine mammal populations.

  6. A survey of Cryptosporidium oocysts in water supplies during a 10-year period (2000-2009) in Seoul.

    PubMed

    Lee, Mok Young; Cho, Eun Joo; Lee, Jin Hyo; Han, Sun Hee; Park, Yong Sang

    2010-09-01

    This study has been conducted to estimate the occurrence of Cryptosporidium oocysts in water supplies in the Metropolitan area of Seoul, South Korea, for 10 years from 2000 to 2009. Water samples were collected quarterly at 6 intakes in the Han River and its largest stream and 6 conventional Water Treatment Plants (WTPs) serving drinking water for 10 million people of Seoul. Cryptosporidium oocysts were found in 22.5% of intake water samples and arithmetic mean was 0.65 oocysts/10 L (range 0-22 oocysts/10 L). Although the annual mean of oocyst number was as low as 0.04-1.90 oocysts/10 L, 3 peaks in 2004 and 2007 were observed and the pollution level was a little higher in winter. The lowest density was observed at Paldang intake and the pollution level increased at Kuui and Jayang intakes. At the end of the largest stream, oocysts were found in 70% of collected samples (mean 5.71 oocysts/10 L) and it seemed that its joining the Han River resulted in the increase at Kuui intake and downstream. Oocyst removal by physical process exceeded 2.0-2.3 log and then all finished water samples collected at 6 WTPs were negative for Cryptosporidium in each 100 L sample for 10 years. These results suggested that domestic wastewater from the urban region could be a source of Cryptosporidium pollution and separating sewage systems adjacent to the intakes could be meaningful for some intakes having weakness related to parasitological water quality.

  7. Prevalence of cryptosporidium infection and characteristics of oocyst shedding in a breeding colony of leopard geckos (Eublepharis macularius).

    PubMed

    Deming, Clare; Greiner, Ellis; Uhl, Elizabeth W

    2008-12-01

    Cryptosporidiosis is an emerging problem in reptile medicine and has been associated with a wasting syndrome in leopard geckos (Eublepharis macularius). This study determined the prevalence of infection in a breeding colony of leopard geckos to be 9.8%. Two groups of 20 geckos, one that was fecal positive for oocysts of Cryptosporidium sp., and one, whose individuals were fecal negative at the inception of the study, were followed for 2 mo. Fecal samples were tested for oocysts every 2 wk, body weights were measured, and a body condition score was assigned for each gecko. Selected geckos from these two groups were euthanized and necropsied. There were statistically significant differences (P < 0.05) between the two groups for mean body weight, mean body condition score, and prevalence of infection. Cryptosporidium sp. infection is endemic in this breeding colony, and there were a large number of geckos with a subclinical or carrier state of infection. These animals continued to be infected with Cryptosporidium sp. but gained weight and remained in good body condition. Only one gecko in the entire group of 40 was confirmed to be negative for oocysts or developmental stages by repeated fecal exams and histopathology. An additional 37 severely emaciated geckos from the breeding colony were euthanized, and all were positive for Cryptosporidium sp. on histopathologic examination of the gastrointestinal tract. The results of this study indicate that although some animals can recover from a clinical infection, if a gecko is severely wasted, it should be euthanized because of the poor prognosis and possible source of infection to other geckos.

  8. Giardia duodenalis and Cryptosporidium occurrence in Australian sea lions (Neophoca cinerea) exposed to varied levels of human interaction

    PubMed Central

    Delport, Tiffany C.; Asher, Amy J.; Beaumont, Linda J.; Webster, Koa N.; Harcourt, Robert G.; Power, Michelle L.

    2014-01-01

    Giardia and Cryptosporidium are amongst the most common protozoan parasites identified as causing enteric disease in pinnipeds. A number of Giardia assemblages and Cryptosporidium species and genotypes are common in humans and terrestrial mammals and have also been identified in marine mammals. To investigate the occurrence of these parasites in an endangered marine mammal, the Australian sea lion (Neophoca cinerea), genomic DNA was extracted from faecal samples collected from wild populations (n = 271) in Southern and Western Australia and three Australian captive populations (n = 19). These were screened using PCR targeting the 18S rRNA of Giardia and Cryptosporidium. Giardia duodenalis was detected in 28 wild sea lions and in seven captive individuals. Successful sequencing of the 18S rRNA gene assigned 27 Giardia isolates to assemblage B and one to assemblage A, both assemblages commonly found in humans. Subsequent screening at the gdh and β-giardin loci resulted in amplification of only one of the 35 18S rRNA positive samples at the β-giardin locus. Sequencing at the β-giardin locus assigned the assemblage B 18S rRNA confirmed isolate to assemblage AI. The geographic distribution of sea lion populations sampled in relation to human settlements indicated that Giardia presence in sea lions was highest in populations less than 25 km from humans. Cryptosporidium was not detected by PCR screening in either wild colonies or captive sea lion populations. These data suggest that the presence of G. duodenalis in the endangered Australian sea lion is likely the result of dispersal from human sources. Multilocus molecular analyses are essential for the determination of G. duodenalis assemblages and subsequent inferences on transmission routes to endangered marine mammal populations. PMID:25426423

  9. Molecular epidemiology of Cryptosporidium spp. and Giardia spp. in mussels (Mytilus californianus) and California sea lions (Zalophus californianus) from Central California.

    PubMed

    Adell, A D; Smith, W A; Shapiro, K; Melli, A; Conrad, P A

    2014-12-01

    Cryptosporidium and Giardia are of public health importance, with recognized transmission through recreational waters. Therefore, both can contaminate marine waters and shellfish, with potential to infect marine mammals in nearshore ecosystems. A 2-year study was conducted to evaluate the presence of Cryptosporidium and Giardia in mussels located at two distinct coastal areas in California, namely, (i) land runoff plume sites and (ii) locations near sea lion haul-out sites, as well as in feces of California sea lions (CSL) (Zalophus californianus) by the use of direct fluorescent antibody (DFA) detection methods and PCR with sequence analysis. In this study, 961 individual mussel hemolymph samples, 54 aliquots of pooled mussel tissue, and 303 CSL fecal samples were screened. Giardia duodenalis assemblages B and D were detected in hemolymph from mussels collected near two land runoff plume sites (Santa Rosa Creek and Carmel River), and assemblages C and D were detected in hemolymph from mussels collected near a sea lion haul-out site (White Rock). These results suggest that mussels are being contaminated by protozoa carried in terrestrial runoff and/or shed in the feces of CSL. Furthermore, low numbers of oocysts and cysts morphologically similar to Cryptosporidium and Giardia, respectively, were detected in CSL fecal samples, suggesting that CSL could be a source and a host of protozoan parasites in coastal environments. The results of this study showed that Cryptosporidium and Giardia spp. from the feces of terrestrial animals and CSL can contaminate mussels and coastal environments.

  10. Modelling the impact of sanitation, population growth and urbanization on human emissions of Cryptosporidium to surface waters—a case study for Bangladesh and India

    NASA Astrophysics Data System (ADS)

    Vermeulen, Lucie C.; de Kraker, Jelske; Hofstra, Nynke; Kroeze, Carolien; Medema, Gertjan

    2015-09-01

    Cryptosporidium is a protozoan parasite that can cause diarrhoea. Human faeces are an important source of Cryptosporidium in surface waters. We present a model to study the impact of sanitation, urbanization and population growth on human emissions of Cryptosporidium to surface waters. We build on a global model by Hofstra et al (2013 Sci. Total Environ. 442 10-9) and zoom into Bangladesh and India as illustrative case studies. The model is most sensitive to changes in oocyst excretion and infection rate, and to assumptions on the share of faeces reaching the surface water for different sanitation types. We find urban centres to be hotspots of human Cryptosporidium emissions. We estimate that 53% (Bangladesh) and 91% (India) of total emissions come from urban areas. 50% of oocysts come from only 8% (Bangladesh) and 3% (India) of the country area. In the future, population growth and urbanization may further deteriorate water quality in Bangladesh and India, despite improved sanitation. Under our ‘business as usual’ (‘sanitation improvements’) scenario, oocyst emissions will increase by a factor 2.0 (1.2) for India and 2.9 (1.1) for Bangladesh between 2010 and 2050. Population growth, urbanization and sanitation development are important processes to consider for large scale water quality modelling.

  11. Molecular Epidemiology of Cryptosporidium spp. and Giardia spp. in Mussels (Mytilus californianus) and California Sea Lions (Zalophus californianus) from Central California

    PubMed Central

    Adell, A. D.; Shapiro, K.; Melli, A.; Conrad, P. A.

    2014-01-01

    Cryptosporidium and Giardia are of public health importance, with recognized transmission through recreational waters. Therefore, both can contaminate marine waters and shellfish, with potential to infect marine mammals in nearshore ecosystems. A 2-year study was conducted to evaluate the presence of Cryptosporidium and Giardia in mussels located at two distinct coastal areas in California, namely, (i) land runoff plume sites and (ii) locations near sea lion haul-out sites, as well as in feces of California sea lions (CSL) (Zalophus californianus) by the use of direct fluorescent antibody (DFA) detection methods and PCR with sequence analysis. In this study, 961 individual mussel hemolymph samples, 54 aliquots of pooled mussel tissue, and 303 CSL fecal samples were screened. Giardia duodenalis assemblages B and D were detected in hemolymph from mussels collected near two land runoff plume sites (Santa Rosa Creek and Carmel River), and assemblages C and D were detected in hemolymph from mussels collected near a sea lion haul-out site (White Rock). These results suggest that mussels are being contaminated by protozoa carried in terrestrial runoff and/or shed in the feces of CSL. Furthermore, low numbers of oocysts and cysts morphologically similar to Cryptosporidium and Giardia, respectively, were detected in CSL fecal samples, suggesting that CSL could be a source and a host of protozoan parasites in coastal environments. The results of this study showed that Cryptosporidium and Giardia spp. from the feces of terrestrial animals and CSL can contaminate mussels and coastal environments. PMID:25281384

  12. Maximum power point tracking algorithm based on sliding mode and fuzzy logic for photovoltaic sources under variable environmental conditions

    NASA Astrophysics Data System (ADS)

    Atik, L.; Petit, P.; Sawicki, J. P.; Ternifi, Z. T.; Bachir, G.; Della, M.; Aillerie, M.

    2017-02-01

    Solar panels have a nonlinear voltage-current characteristic, with a distinct maximum power point (MPP), which depends on the environmental factors, such as temperature and irradiation. In order to continuously harvest maximum power from the solar panels, they have to operate at their MPP despite the inevitable changes in the environment. Various methods for maximum power point tracking (MPPT) were developed and finally implemented in solar power electronic controllers to increase the efficiency in the electricity production originate from renewables. In this paper we compare using Matlab tools Simulink, two different MPP tracking methods, which are, fuzzy logic control (FL) and sliding mode control (SMC), considering their efficiency in solar energy production.

  13. The European rabbit (Oryctolagus cuniculus), a source of zoonotic cryptosporidiosis.

    PubMed

    Robinson, G; Chalmers, R M

    2010-12-01

    Cryptosporidium spp. have been found in the faeces of over 150 mammalian host species, but the risks to public health from wildlife are poorly understood. In summer 2008, the Cryptosporidium sp. rabbit genotype was identified as the aetiological agent in an outbreak of waterborne human cryptosporidiosis. The source was a wild rabbit that had entered a treated water tank. To establish current knowledge about Cryptosporidium spp. infecting lagomorphs, especially the host range and biological characteristics of the rabbit genotype, and the potential risks to public health that rabbits may pose in the transmission of zoonotic cryptosporidiosis, we undertook a literature and data review. The literature returned demonstrates that although the European rabbit (Oryctolagus cuniculus) has been the most widely studied lagomorph, few large scale studies were found. The prevalence of Cryptosporidium spp. in wild rabbit populations in the two large scale studies was 0.9% (95%CI 0.2-5.0) and 0.0% (95%CI 0.0-1.6). Neither study provided age nor sex profiles nor typing of Cryptosporidium isolates. The infecting Cryptosporidium species was confirmed in just four other studies of rabbits, all of which showed the rabbit genotype. Human-infectious Cryptosporidium species including Cryptosporidium parvum have caused experimental infections in rabbits and it is likely that this may also occur naturally. No published studies of the host range and biological features of the Cryptosporidium rabbit genotype were identified, but information was generated on the identification and differentiation of the rabbit genotype at various genetic loci. Both pet and wild rabbits are a potential source of human cryptosporidiosis and as such, good hygiene practices are recommended during and after handling rabbits or exposure to their faeces, or potentially contaminated surfaces. Water supplies should be protected against access by wildlife, including rabbits.

  14. VideoHacking: Automated Tracking and Quantification of Locomotor Behavior with Open Source Software and Off-the-Shelf Video Equipment.

    PubMed

    Conklin, Emily E; Lee, Kathyann L; Schlabach, Sadie A; Woods, Ian G

    2015-01-01

    Differences in nervous system function can result in differences in behavioral output. Measurements of animal locomotion enable the quantification of these differences. Automated tracking of animal movement is less labor-intensive and bias-prone than direct observation, and allows for simultaneous analysis of multiple animals, high spatial and temporal resolution, and data collection over extended periods of time. Here, we present a new video-tracking system built on Python-based software that is free, open source, and cross-platform, and that can analyze video input from widely available video capture devices such as smartphone cameras and webcams. We validated this software through four tests on a variety of animal species, including larval and adult zebrafish (Danio rerio), Siberian dwarf hamsters (Phodopus sungorus), and wild birds. These tests highlight the capacity of our software for long-term data acquisition, parallel analysis of multiple animals, and application to animal species of different sizes and movement patterns. We applied the software to an analysis of the effects of ethanol on thigmotaxis (wall-hugging) behavior on adult zebrafish, and found that acute ethanol treatment decreased thigmotaxis behaviors without affecting overall amounts of motion. The open source nature of our software enables flexibility, customization, and scalability in behavioral analyses. Moreover, our system presents a free alternative to commercial video-tracking systems and is thus broadly applicable to a wide variety of educational settings and research programs.

  15. North American tree squirrels and ground squirrels with overlapping ranges host different Cryptosporidium species and genotypes.

    PubMed

    Stenger, Brianna L S; Clark, Mark E; Kváč, Martin; Khan, Eakalak; Giddings, Catherine W; Prediger, Jitka; McEvoy, John M

    2015-12-01

    Wildlife-associated Cryptosporidium are an emerging cause of cryptosporidiosis in humans. The present study was undertaken to determine the extent to which North American tree squirrels and ground squirrels host zoonotic Cryptosporidium species and genotypes. Fragments of the Cryptosporidium 18S rRNA and actin genes were amplified and sequenced from fecal samples obtained from three tree squirrel and three ground squirrel species. In tree squirrels, Cryptosporidium was identified in 40.5% (17/42) of American red squirrels (Tamiasciurus hudsonicus), 40.4% (55/136) of eastern gray squirrels (Sciurus carolinensis), and 28.6% (2/7) of fox squirrels (Sciurus niger). Human-pathogenic Cryptosporidium ubiquitum and Cryptosporidium skunk genotype were the most prevalent species/genotypes in tree squirrels. Because tree squirrels live in close proximity to humans and are frequently infected with potentially zoonotic Cryptosporidium species/genotypes, they may be a significant reservoir of infection in humans. In ground squirrels, Cryptosporidium was detected in 70.2% (33/47) of 13-lined ground squirrels (Ictidomys tridecemlineatus), 35.1% (27/77) of black-tailed prairie dogs (Cynomys ludovicianus), and the only golden-mantled ground squirrel (Callospermophilus lateralis) that was sampled. Cryptosporidium rubeyi and ground squirrel genotypes I, II, and III were identified in isolates from these ground squirrel species. In contrast to the Cryptosporidium infecting tree squirrels, these species/genotypes appear to be specific for ground squirrels and are not associated with human disease.

  16. CRYPTOSPORIDIUM VIRULENCE DETERMINANTS-ARE WE THERE YET? (R828035)

    EPA Science Inventory

    Exposure to Cryptosporidium parvum in healthy individuals results in transient infection that may be asymptomatic or can result in self-limited diarrhoea. In contrast, acquired immune deficiency syndrome patients with cryptosporidiosis can experience severe manifestatio...

  17. Cryptosporidium hominis n. sp. (Apicomplexa: Cryptosporidiidae) from Homo sapiens.

    PubMed

    Morgan-Ryan, Una M; Fall, Abbie; Ward, Lucy A; Hijjawi, Nawal; Sulaiman, Irshad; Fayer, Ronald; Thompson, R C Andrew; Olson, M; Lal, Altaf; Xiao, Lihua

    2002-01-01

    The structure and infectivity of the oocysts of a new species of Cryptosporidium from the feces of humans are described. Oocysts are structurally indistinguishable from those of Cryptosporidium parvum. Oocysts of the new species are passed fully sporulated, lack sporocysts. and measure 4.4-5.4 microm (mean = 4.86) x 4.4-5.9 microm (mean = 5.2 microm) with a length to width ratio 1.0-1.09 (mean 1.07) (n = 100). Oocysts were not infectious for ARC Swiss mice, nude mice. Wistar rat pups, puppies, kittens or calves, but were infectious to neonatal gnotobiotic pigs. Pathogenicity studies in the gnotobiotic pig model revealed significant differences in parasite-associated lesion distribution (P = 0.005 to P = 0.02) and intensity of infection (P = 0.04) between C. parvum and this newly described species from humans. In vitro cultivation studies have also revealed growth differences between the two species. Multi-locus analysis of numerous unlinked loci, including a preliminary sequence scan of the entire genome demonstrated this species to be distinct from C. parvum and also demonstrated a lack of recombination, providing further support for its species status. Based on biological and molecular data, this Cryptosporidium infecting the intestine of humans is proposed to be a new species Cryptosporidium hominis n. sp.

  18. Prevalence of Cryptosporidium in small ruminants from Veracruz, Mexico

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryptosporidiosis is a zoonotic disease caused by the protozoan parasite Cryptosporidium spp. that can affect domestic animal and human populations. In newborn ruminants, cryptosporidiosis is characterized by outbreaks of diarrhea, which can result in high morbidity and economic impact. The aim of t...

  19. Subtyping Cryptosporidium ubiquitum, an emerging zoonotic pathogen in humans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryptosporidium ubiquitum is an emerging and important human pathogen with a broad range of mammalian hosts. Linkage between human and animal cases of C. ubiquitum has not been possible because of a lack of subtyping tools. In this study, we utilized the 60 kDa glycoprotein gene of C. ubiquitum to d...

  20. CONTROL OF CRYPTOSPORIDIUM OOCYSTS BY STEADY-STATE CONVENTIONAL TREATMENT

    EPA Science Inventory

    Pilot-scale experiments have been performed to assess the ability of conventional treatment to control Cryptosporidium oocysts under steady-state conditions. The work was performed with a pilot plant that was designed to minimize flow rates and, as a result, the number of oocyst...

  1. Genetic modification of the diarrhoeal pathogen Cryptosporidium parvum.

    PubMed

    Vinayak, Sumiti; Pawlowic, Mattie C; Sateriale, Adam; Brooks, Carrie F; Studstill, Caleb J; Bar-Peled, Yael; Cipriano, Michael J; Striepen, Boris

    2015-07-23

    Recent studies into the global causes of severe diarrhoea in young children have identified the protozoan parasite Cryptosporidium as the second most important diarrhoeal pathogen after rotavirus. Diarrhoeal disease is estimated to be responsible for 10.5% of overall child mortality. Cryptosporidium is also an opportunistic pathogen in the contexts of human immunodeficiency virus (HIV)-caused AIDS and organ transplantation. There is no vaccine and only a single approved drug that provides no benefit for those in gravest danger: malnourished children and immunocompromised patients. Cryptosporidiosis drug and vaccine development is limited by the poor tractability of the parasite, which includes a lack of systems for continuous culture, facile animal models, and molecular genetic tools. Here we describe an experimental framework to genetically modify this important human pathogen. We established and optimized transfection of C. parvum sporozoites in tissue culture. To isolate stable transgenics we developed a mouse model that delivers sporozoites directly into the intestine, a Cryptosporidium clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system, and in vivo selection for aminoglycoside resistance. We derived reporter parasites suitable for in vitro and in vivo drug screening, and we evaluated the basis of drug susceptibility by gene knockout. We anticipate that the ability to genetically engineer this parasite will be transformative for Cryptosporidium research. Genetic reporters will provide quantitative correlates for disease, cure and protection, and the role of parasite genes in these processes is now open to rigorous investigation.

  2. Genetic characterization of Cryptosporidium spp. among patients with gastrointestinal complaints

    PubMed Central

    Ranjbar, Reza; Baghaei, Kaveh; Nazemalhosseini Mojarad, Ehsan

    2016-01-01

    Aim: This study investigated subtypes of Cryptosporidium in patients with gastrointestinal complaints in Tehran, Iran. Background: Cryptosporidium, an intracellular protozean parasite, is among the major causative agents of gastroenteritis disorders in humans. It also causes water-borne and food-borne outbreaks of diarrheal diseases. Patients and methods: A total of 1685 fecal samples were collected from patients with gastrointestinal complaints who had been referred to clinical laboratories Tehran, Iran. The primary diagnosis was established by the detection of oocysts using the modified Ziehl-Neelsen staining method and following that, the positive microscopically samples were selected for sequence analysis of the partial 60 kDa glycoprotein (gp60) gene. Results: Out of 1685 collected samples, 7 (0.4 %) were positive for Cryptosporidium oocysts. Sequence analysis of gp60 gene in seven Cryptosporidium isolates revealed that two subtype families were identified, IIa and IId. Five (of 7) isolates belonged to the subtype family IIa and the remaining two isolates belonged to IId. Two sub-types were recognized within the subtype family II,a including IIaA16G2R1 (3/5), IIaA17G1R1 (2/5), while IIdA17G1d was the only subtype within IId subtype family. Conclusion: The predominance of zoonotic subtype families of C. parvum species (IIa, IId) in this study highlights the importance of zoonotic transmission of cryptosporidiosis in the country. PMID:27895856

  3. TESTING METHODS FOR DETECTION OF CRYPTOSPORIDIUM SPP. IN WATER SAMPLES

    EPA Science Inventory

    A large waterborne outbreak of cryptosporidiosis in Milwaukee, Wisconsin, U.S.A. in 1993 prompted a search for ways to prevent large-scale waterborne outbreaks of protozoan parasitoses. Methods for detecting Cryptosporidium parvum play an integral role in strategies that lead to...

  4. Identification of Cryptosporidium from Dairy Cattle in Pahang, Malaysia

    PubMed Central

    Hisamuddin, Nur Hazirah; Hashim, Najat; Soffian, Sharmeen Nellisa; Amin, Mohd Hishammfariz Mohd; Wahab, Ridhwan Abdul; Mohammad, Mardhiah; Isa, Muhammad Lokman Md; Yusof, Afzan Mat

    2016-01-01

    Cryptosporidium, a protozoan parasite, can cause cryptosporidiosis which is a gastrointestinal disease that can infect humans and livestock. Cattle are the most common livestock that can be infected with this protozoan. This study was carried out to determine the prevalence of Cryptosporidium infection in cattle in Kuantan, Pahang, Malaysia and to find out the association between the occurrence of infection and 3 different ages of cattle (calves less than 1 year, yearling, and adult cattle). The samples were processed by using formol-ether concentration technique and stained by modified Ziehl Neelsen. The results showed that 15.9% (24/151) of cattle were positive for Cryptosporidium oocysts. The occurrence of Cryptosporidium in calves less than 1 year was the highest with the percentage of 20.0% (11/55) followed by yearling and adult cattle, with the percentage occurrence of 15.6 % (7/45) and 11.8% (6/51), respectively. There was no significant association between the occurrence and age of cattle and presence of diarrhea. Good management practices and proper hygiene management must be taken in order to reduce the infection. It is highly important to control the infection since infected cattle may serve as potential reservoirs of the infection to other animals and humans, especially animal handlers. PMID:27180579

  5. Molecular detection of Cryptosporidium cuniculus in rabbits in Australia.

    PubMed

    Nolan, Matthew J; Jex, Aaron R; Haydon, Shane R; Stevens, Melita A; Gasser, Robin B

    2010-12-01

    In the United Kingdom, rabbits have been reported to harbour genotypes of Cryptosporidium (now recognized as C. cuniculus) identical to those from human patients exhibiting symptoms of cryptosporidiosis. The high density of rabbits in many regions of Australia, including both rural and urban as well as natural water catchments areas, and the absence of any information on Cryptosporidium from lagomorphs in this country stimulated the present study. We undertook an epidemiological study that genetically characterized Cryptosporidium from rabbits from four locations in Victoria by PCR-coupled sequencing and phylogenetic analysis of sequence data for loci within the small subunit of nuclear ribosomal RNA (SSU; for specific identification) and the 60kDa glycoprotein gene (gp60; for genotypic/subgenotypic identification). Cryptosporidium was detected in 12 (6.8%) of 176 individual faecal samples. For SSU, all 12 sequences were identical to each other and to that of C. cuniculus. For pgp60, all corresponding sequences matched the known genotype Vb, and were classified as subgenotype VbA23R3 (n=11) and VbA26R4 (n=1), which are both new records. Present evidence indicates that genotype Vb is limited to rabbits; however, it would be premature to conclude that this genotype is not zoonotic. Future studies should focus on the zoonotic potential of C. cuniculus from rabbits and a wide range of yet unstudied animals. (Nucleotide sequences reported in this paper are available in the GenBank database under accession nos. HM852431-HM852433).

  6. ENHANCED DAPI STAINING FOR CRYPTOSPORIDIUM IN WATER SAMPLES

    EPA Science Inventory

    The U.S. Environmental Protection Agency's Method 1623 is used to detect and quantify the presence of {ital Cryptosporidium} spp. oocysts in water. The protocol consists of concentrating a sample, staining this concentrate with a fluorescent antibody, and examining the sample mi...

  7. A sensitive method for detecting and genotyping Cryptosporidium parvum oocysts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryptosporidium parvum oocysts represent a considerable health risk to humans and animals because the parasite has a low infectious dose and usually exists in low numbers in environmental samples, which makes detection problematic. The purpose of this study was to evaluate Cryspovirus as a target f...

  8. Chlorine disinfection of recreational water for Cryptosporidium parvum.

    PubMed Central

    Carpenter, C.; Fayer, R.; Trout, J.; Beach, M. J.

    1999-01-01

    We examined the effects of chlorine on oocyst viability, under the conditions of controlled pH and elevated calcium concentrations required for most community swimming pools. We found that fecal material may alter the Ct values (chlorine concentration in mg/L, multiplied by time in minutes) needed to disinfect swimming pools or other recreational water for Cryptosporidium parvum. PMID:10458969

  9. Molecular epidemiology of Cryptosporidium and Giardia in cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryptosporidium spp. and Giardia duodenalis are enteric protozoan parasites that infect a wide range of vertebrate hosts including humans. Infections with both parasites are known as one of the most common causes of diarrhea in humans and livestock. The epidemiology of cryptosporidiosis and giardias...

  10. Prevalence and species of Cryptosporidium in human populations worldwide

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A high prevalence of cryptosporidiosis has been reported in children under 5 years of age in sub-Saharan Africa, especially, but not exclusively, the malnourished and those positive for HIV infection. Infection rates are highest during the rainy season. Cryptosporidium hominis, which is transmitted ...

  11. Presence of Cryptosporidium spp. and Giardia duodenalis through drinking water.

    PubMed

    Castro-Hermida, José Antonio; García-Presedo, Ignacio; Almeida, André; González-Warleta, Marta; Correia Da Costa, José Manuel; Mezo, Mercedes

    2008-11-01

    To evaluate the presence of Cryptosporidium spp. and Giardia duodenalis in the influent and final effluent of sixteen drinking water treatment plants located in a hydrographic basin in Galicia (NW Spain) - in which the principal river is recognised as a Site of Community Importance (SCI) - estimate the efficiency of treatment plants in removing these protozoans and determine the species and genotypes of the parasites by means of a molecular assay. All plant samples of influent and final effluent (50-100 l) were examined in the spring, summer, autumn and winter of 2007. A total of 128 samples were analysed by method 1623, developed by US Environmental Protection Agency for isolation and detection of both parasites. To identify the genotypes present the following genes were amplified and sequenced: 18S SSU rRNA (Cryptosporidium spp.) and b-giardina (G. duodenalis). The mean concentrations of parasites in the influent were 0.0-10.5 Cryptosporidium spp. oocysts per litre and 1.0-12.8 of G. duodenalis cysts per litre. In the final treated effluent, the mean concentration of parasites ranged from 0.0-3.0 oocysts per litre and 0.5-4.0 cysts per litre. The distribution of results by season revealed that in all plants, the highest numbers of (oo)cysts were recorded in spring and summer. Cryptosporidium parvum, C. andersoni, C. hominis and assemblages A-I, A-II, E of G. duodenalis were detected. Cryptosporidium spp. and G. duodenalis were consistently found at high concentrations in drinking water destined for human and animal consumption in the hydrographic basin under study, in Galicia (NW Spain). It is important that drinking water treatment authorities rethink the relevance of contamination levels of both parasites in drinking water and develop adequate countermeasures.

  12. A Modified EPA Method 1623 that Uses Tangential Flow Hollow-fiber Ultrafiltration and Heat Dissociation Steps to Detect Waterborne Cryptosporidium and Giardia spp.

    PubMed Central

    Rhodes, Eric R.; Villegas, Leah Fohl; Shaw, Nancy J.; Miller, Carrie; Villegas, Eric N.

    2012-01-01

    Cryptosporidium and Giardia species are two of the most prevalent protozoa that cause waterborne diarrheal disease outbreaks worldwide. To better characterize the prevalence of these pathogens, EPA Method 1623 was developed and used to monitor levels of these organisms in US drinking water supplies 12. The method has three main parts; the first is the sample concentration in which at least 10 L of raw surface water is filtered. The organisms and trapped debris are then eluted from the filter and centrifuged to further concentrate the sample. The second part of the method uses an immunomagnetic separation procedure where the concentrated water sample is applied to immunomagnetic beads that specifically bind to the Cryptosporidium oocysts and Giardia cysts allowing for specific removal of the parasites from the concentrated debris. These (oo)cysts are then detached from the magnetic beads by an acid dissociation procedure. The final part of the method is the immunofluorescence staining and enumeration where (oo)cysts are applied to a slide, stained, and enumerated by microscopy. Method 1623 has four listed sample concentration systems to capture Cryptosporidium oocysts and Giardia cysts in water: Envirochek filters (Pall Corporation, Ann Arbor, MI), Envirochek HV filters (Pall Corporation), Filta-Max filters (IDEXX, Westbrook, MA), or Continuous Flow Centrifugation (Haemonetics, Braintree, MA). However, Cryptosporidium and Giardia (oo)cyst recoveries have varied greatly depending on the source water matrix and filters used1,14. A new tangential flow hollow-fiber ultrafiltration (HFUF) system has recently been shown to be more efficient and more robust at recovering Cryptosporidium oocystsand Giardia cysts from various water matrices; moreover, it is less expensive than other capsule filter options and can concentrate multiple pathogens simultaneously1-3,5-8,10,11. In addition, previous studies by Hill and colleagues demonstrated that the HFUF significantly improved

  13. Identification and genotyping of Giardia spp. and Cryptosporidium spp. isolates in aquatic birds in the Salburua wetlands, Álava, Northern Spain.

    PubMed

    Cano, Lourdes; de Lucio, Aida; Bailo, Begoña; Cardona, Guillermo A; Muadica, Aly Salimo Omar; Lobo, Luis; Carmena, David

    2016-05-15

    Aquatic birds are known to be suitable hosts for a number of avian-specific species and genotypes of the enteric protozoan parasites Giardia and Cryptosporidium. Waterbirds have also been reported as sporadic carriers of species of both pathogens from human or domestic animal origin via environmental contamination. Because aquatic birds can shed substantial amounts of infective Giardia and Cryptosporidium (oo)cysts to the environment including surface waters intended for human consumption, this situation may pose a potential risk of waterborne zoonotic disease. A total of 265 waterbird faecal samples were collected from May 2014 to June 2015 at Salburua (Álava), one of the most valued continental wetlands in northern Spain. The detection of Giardia oocyst and Cryptosporidium oocysts was carried out by direct fluorescence microscopy and molecular (PCR and sequence analysis) methods targeting the small subunit ribosomal RNA gene of the parasites. Typing of Giardia duodenalis isolates at the sub-assemblage level was based on the specific amplification and sequencing of a partial fragment of the glutamate dehydrogenase gene. Overall, Giardia cysts and Cryptosporidium oocysts were detected in 22 (8.3%) and 6 (2.3%), respectively, of the 265 faecal samples analysed. The two only Giardia isolates characterized (one novel, one known) were assigned to the sub-assemblage BIV of G. duodenalis, none of them previously reported in Spanish human isolates. This finding raises doubts about the actual origin of the infection and whether waterbirds may serve as potential source of infective Giardia cysts to humans via waterborne transmission or through direct contact. The six Cryptosporidium isolates obtained were characterized as avian genotype III (n=4), duck genotype b (n=1), and goose genotype Id (n=1), all considered avian-specific and therefore of negligible risk of zoonotic infection.

  14. Detection of Cryptosporidium sp. in two new seal species, Phoca vitulina and Cystophora cristata, and a novel Cryptosporidium genotype in a third seal species, Pagophilus groenlandicus, from the Gulf of Maine.

    PubMed

    Bass, A L; Wallace, C C; Yund, P O; Ford, T E

    2012-04-01

    Data on the geographic distribution and host specificity of Cryptosporidium spp. are critical for developing an understanding of likely transmission patterns in nature. During a molecular-based survey of fecal samples from 293 terrestrial and aquatic animals in Maine, USA, we detected Cryptosporidium sp. in 11 harbor seals (Phoca vitulina), 1 hooded seal (Cystophora cristata), and 1 harp seal (Pagophilus groenlandicus). None of the terrestrial or freshwater mammal fecal samples or bird samples tested positive for Cryptosporidium sp. However, the sequencing results of the small subunit (ssu) rRNA gene indicate that the seals were infected with an undescribed species of Cryptosporidium , previously isolated only from ringed seals (Phoca hispida) in northern Quebec, Canada. In addition, the Cryptosporidium sp. detected in the harp seal is significantly different from the previously observed Cryptosporidium sp. in other seals. We confirmed the genetic distinctiveness of this Cryptosporidium genotype and the identity of the other Cryptosporidium sp. seal ssu rRNA sequences by using data from the 70-kDa heat shock protein gene. Based on phylogenetic reconstructions of both genes, it seems that either Cryptosporidium canis or C. felis are sister species to the seal associated Cryptosporidium spp. Our findings extend the range of " Cryptosporidium sp. seal" well south of the 55th parallel, add other species to the list of seals affected by Cryptosporidium sp., and highlight the presence of unrecognized population and potentially species level variation in Cryptosporidium.

  15. Cryptosporidium proliferans n. sp. (Apicomplexa: Cryptosporidiidae): Molecular and Biological Evidence of Cryptic Species within Gastric Cryptosporidium of Mammals

    PubMed Central

    Kváč, Martin; Havrdová, Nikola; Hlásková, Lenka; Daňková, Tereza; Kanděra, Jiří; Ježková, Jana; Vítovec, Jiří; Sak, Bohumil; Ortega, Ynes; Xiao, Lihua; Modrý, David; Chelladurai, Jeba Rose Jennifer Jesudoss; Prantlová, Veronika; McEvoy, John

    2016-01-01

    The morphological, biological, and molecular characteristics of Cryptosporidium muris strain TS03 are described, and the species name Cryptosporidium proliferans n. sp. is proposed. Cryptosporidium proliferans obtained from a naturally infected East African mole rat (Tachyoryctes splendens) in Kenya was propagated under laboratory conditions in rodents (SCID mice and southern multimammate mice, Mastomys coucha) and used in experiments to examine oocyst morphology and transmission. DNA from the propagated C. proliferans isolate, and C. proliferans DNA isolated from the feces of an African buffalo (Syncerus caffer) in Central African Republic, a donkey (Equus africanus) in Algeria, and a domestic horse (Equus caballus) in the Czech Republic were used for phylogenetic analyses. Oocysts of C. proliferans are morphologically distinguishable from C. parvum and C. muris HZ206, measuring 6.8–8.8 (mean = 7.7 μm) × 4.8–6.2 μm (mean = 5.3) with a length to width ratio of 1.48 (n = 100). Experimental studies using an isolate originated from T. splendens have shown that the course of C. proliferans infection in rodent hosts differs from that of C. muris and C. andersoni. The prepatent period of 18–21 days post infection (DPI) for C. proliferans in southern multimammate mice (Mastomys coucha) was similar to that of C. andersoni and longer than the 6–8 DPI prepatent period for C. muris RN66 and HZ206 in the same host. Histopatologicaly, stomach glands of southern multimammate mice infected with C. proliferans were markedly dilated and filled with necrotic material, mucus, and numerous Cryptosporidium developmental stages. Epithelial cells of infected glands were atrophic, exhibited cuboidal or squamous metaplasia, and significantly proliferated into the lumen of the stomach, forming papillary structures. The epithelial height and stomach weight were six-fold greater than in non-infected controls. Phylogenetic analyses based on small subunit rRNA, Cryptosporidium oocyst

  16. Cryptosporidium proliferans n. sp. (Apicomplexa: Cryptosporidiidae): Molecular and Biological Evidence of Cryptic Species within Gastric Cryptosporidium of Mammals.

    PubMed

    Kváč, Martin; Havrdová, Nikola; Hlásková, Lenka; Daňková, Tereza; Kanděra, Jiří; Ježková, Jana; Vítovec, Jiří; Sak, Bohumil; Ortega, Ynes; Xiao, Lihua; Modrý, David; Chelladurai, Jeba Rose Jennifer Jesudoss; Prantlová, Veronika; McEvoy, John

    2016-01-01

    The morphological, biological, and molecular characteristics of Cryptosporidium muris strain TS03 are described, and the species name Cryptosporidium proliferans n. sp. is proposed. Cryptosporidium proliferans obtained from a naturally infected East African mole rat (Tachyoryctes splendens) in Kenya was propagated under laboratory conditions in rodents (SCID mice and southern multimammate mice, Mastomys coucha) and used in experiments to examine oocyst morphology and transmission. DNA from the propagated C. proliferans isolate, and C. proliferans DNA isolated from the feces of an African buffalo (Syncerus caffer) in Central African Republic, a donkey (Equus africanus) in Algeria, and a domestic horse (Equus caballus) in the Czech Republic were used for phylogenetic analyses. Oocysts of C. proliferans are morphologically distinguishable from C. parvum and C. muris HZ206, measuring 6.8-8.8 (mean = 7.7 μm) × 4.8-6.2 μm (mean = 5.3) with a length to width ratio of 1.48 (n = 100). Experimental studies using an isolate originated from T. splendens have shown that the course of C. proliferans infection in rodent hosts differs from that of C. muris and C. andersoni. The prepatent period of 18-21 days post infection (DPI) for C. proliferans in southern multimammate mice (Mastomys coucha) was similar to that of C. andersoni and longer than the 6-8 DPI prepatent period for C. muris RN66 and HZ206 in the same host. Histopatologicaly, stomach glands of southern multimammate mice infected with C. proliferans were markedly dilated and filled with necrotic material, mucus, and numerous Cryptosporidium developmental stages. Epithelial cells of infected glands were atrophic, exhibited cuboidal or squamous metaplasia, and significantly proliferated into the lumen of the stomach, forming papillary structures. The epithelial height and stomach weight were six-fold greater than in non-infected controls. Phylogenetic analyses based on small subunit rRNA, Cryptosporidium oocyst wall

  17. Measurement of angular distribution of neutron flux for the 6MeV race-track microtron based pulsed neutron source.

    PubMed

    Patil, B J; Chavan, S T; Pethe, S N; Krishnan, R; Dhole, S D

    2010-09-01

    The 6MeV race track microtron based pulsed neutron source has been designed specifically for the elemental analysis of short lived activation products, where the low neutron flux requirement is desirable. Electrons impinges on a e-gamma target to generate bremsstrahlung radiations, which further produces neutrons by photonuclear reaction in gamma-n target. The optimisation of these targets along with their spectra were estimated using FLUKA code. The measurement of neutron flux was carried out by activation of vanadium at different scattering angles. Angular distribution of neutron flux indicates that the flux decreases with increase in the angle and are in good agreement with the FLUKA simulation.

  18. Cryptosporidium species and Cryptosporidium parvum subtypes in dairy calves and goat kids reared under traditional farming systems in Turkey.

    PubMed

    Taylan-Ozkan, Aysegul; Yasa-Duru, Sibel; Usluca, Selma; Lysen, Colleen; Ye, Jianbin; Roellig, Dawn M; Feng, Yaoyu; Xiao, Lihua

    2016-11-01

    Molecular characterizations of Cryptosporidium spp. in ruminants reared under traditional animal management systems are scarce and studies conducted thus far have revealed largely an absence of the pathogenic and zoonotic species Cryptosporidium parvum in pre-weaned animals. In this study, we examined Cryptosporidium species and subtype distribution in free-range pre-weaned dairy calves and goat kids with diarrhea. Cryptosporidium-positive specimens from pre-weaned calves on 10 farms and goat kids on 4 farms in Ankara, Balikesir, Corum, Kirikkale, and Kirsehir Provinces, Turkey were genotyped by PCR-restriction length polymorphism analysis of the small subunit rRNA gene, which identified C. parvum in 27 calves and 9 goat kids and Cryptosporidium ryanae in 1 calf. Among the C. parvum isolates successfully subtyped by DNA sequence analysis of the 60 kDa glycoprotein gene, three subtypes were detected in calves, including IIaA13G2R1 (20/23), IIdA18G1 (2/23), and IIdA20G1b (1/23), and four subtypes were detected in goat kids, including IIaA13G2R1 (3/8), IIaA15G1R1 (2/8), IIdA22G1 (2/8), and IIdA18G1 (1/8). Data of the study suggest that dairy calves reared in a traditional cow-calf system in Turkey are mainly infected with a C. parvum subtype rarely seen elsewhere, whereas goat kids are infected with diverse subtypes. As all five C. parvum subtypes found in this study are known human pathogens, pre-weaned farm animals could play a potential role in the transmission of human cryptosporidiosis.

  19. Characterization of saccharides and other organic compounds in fine particles and the use of saccharides to track primary biologically derived carbon sources

    NASA Astrophysics Data System (ADS)

    Jia, Yuling; Bhat, Shagun; Fraser, Matthew P.

    2010-02-01

    A total of 134 aerosol samples ( dp < 2.5 μm) were collected at one rural site and one urban site in Texas from November 2005 to July 2006 to investigate the different sources that contribute to the ambient levels of different compounds. In particular, saccharide compounds were studied as potential tracers to track aerosols of biologically derived origin. The ambient concentration, seasonal variation, and urban/rural comparison of major saccharides and other organic compounds including normal alkanes, hopanes, and carboxylic acids were determined and analyzed relative to characterizing sources of PM2.5. Saccharides, together with other known molecular markers, were analyzed by a positive matrix factorization model and eight source factors were isolated that provide meaningful interpretation of aerosol sources. Three isolated factors were characterized by the dominance of different saccharide compounds and were attributed to wood smoke, sucrose rich bio-aerosols, and fungal spore derived bio-aerosols. It was estimated that wood smoke and primary biologically derived carbon sources contributed 22% and 14% to the measured ambient PM2.5 mass at San Augustine and 16% and 5% to the measured ambient PM2.5 mass at Dallas. The relative PM contribution from other resolved sources were also calculated.

  20. Detection of the Cryptosporidium parvum "human" genotype in a dugong (Dugong dugon).

    PubMed

    Morgan, U M; Xiao, L; Hill, B D; O'Donoghue, P; Limor, J; Lal, A; Thompson, R C

    2000-12-01

    The Cryptosporidium "human" genotype was identified in a paraffin-embedded tissue section from a dugong (Dugong dugon) by 2 independent laboratories. DNA sequencing and polymerase chain reaction/restriction fragment length polymorphism analysis of the 18S ribosomal RNA gene and the acetyl CoA synthethase gene clearly identified the genotype as that of the Cryptosporidium variant that infects humans. This is the first report of the human Cryptosporidium genotype in a nonprimate host.

  1. INVESTIGATION ON CRYPTOSPORIDIUM INFECTIONS IN WILD ANIMALS IN A ZOO IN ANHUI PROVINCE.

    PubMed

    Gu, Youfang; Wang, Xiaolong; Zhou, Cefan; Li, Peiying; Xu, Qianming; Zhao, Changcheng; Liu, Wei; Xu, Wenlong

    2016-09-01

    To assess Cryptosporidium infections among wild animals in a zoo located in Anhui province, we conducted an investigation on the fecal samples collected from 44 primates, 41 herbivores, 44 carnivores and omnivores, and 103 birds in the zoo with the use of Sheather's sugar flotation technique and modified acid-fast staining. Cryptosporidium oocysts were detected in the fecal samples from six primates, two herbivores, four carnivores and omnivores, and seven birds by using Sheather's sugar flotation technique; the prevalence of Cryptosporidium infection in primates, herbivores, carnivores and omnivores and birds was 13.64, 4.88, 9.09, and 6.80%, respectively. Modified acid-fast staining detected the presence of Cryptosporidium oocysts in the fecal samples of one primate, three herbivores, 0 carnivores and omnivores, and one bird, and the prevalence of Cryptosporidium infection in primates, herbivores, carnivores and omnivores and birds was 2.27, 7.32, 0.00, and 0.97%, respectively. Polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) and phylogenetic analysis with the use of the neighbor-joining (NJ) method based on the aligned partial small-subunit (SSU) rRNA gene sequences showed that the protozoan pathogen isolated from primates was Cryptosporidium hominis and the pathogen isolated from camels ( Camelus dromedarius ) was Cryptosporidium andersoni. Subtyping the Cryptosporidium hominis by 60-kDa glycoprotein (GP60) gene phylogenetic analysis showed the Cryptosporidium hominis belongs to the subtype IdA and IbA.

  2. Molecular characterisation of a disseminated Cryptosporidium infection in a Koi carp (Cyprinus carpio).

    PubMed

    Yang, Rongchang; Dorrestein, Gerry M; Ryan, Una

    2016-08-15

    Cryptosporidium is a protozoan parasite that infects a wide range of hosts, yet relatively little is known about the epidemiology of cryptosporidiosis in fish. Here we report a disseminated Cryptosporidium infection in a male Koi carp (Cyprinus carpio), with parasite stages identified deep within the epithelium of the intestine, kidneys, spleen, liver and gills causing severe granulomatous inflammatory lesions. Molecular characterization at two loci; 18S ribosomal RNA (rRNA) and actin, revealed this to be a novel Cryptosporidium genotype, most closely related to Cryptosporidium molnari.

  3. Molecular characterization of Cryptosporidium spp. from fecal samples of birds kept in captivity in Brazil.

    PubMed

    Nakamura, Alex Akira; Simões, Daniel Castendo; Antunes, Rômulo Godik; da Silva, Deuvânia Carvalho; Meireles, Marcelo Vasconcelos

    2009-12-03

    The aim of this study was to determine the prevalence of Cryptosporidium species and genotypes in birds kept in captivity in Brazil. A total of 966 samples from 18 families of birds was collected and stored in 5% potassium dichromate solution at 4 degrees C until processing. Oocysts were purified in Sheather sugar solution following extraction of genomic DNA. Molecular analyses were performed using nested-PCR for amplification of fragments of the 18S subunit of rRNA gene and of the actin gene. Amplification of Cryptosporidium DNA fragments was obtained in 47 (4.86%) samples. Sequencing of amplified fragments and phylogenetic analyses allowed the identification of Cryptosporidium baileyi in a black vulture (Coragyps atratus), a domestic chicken (Gallus gallus domesticus) and a saffron finch (Sicalis flaveola); Cryptosporidium galli in canaries (Serinus canaria), a cockatiel (Nymphicus hollandicus) and lesser seed-finches (Oryzoborus angolensis); Cryptosporidium meleagridis in a domestic chicken (G. g. domesticus); Cryptosporidium parvum in a cockatiel (N. hollandicus); Cryptosporidium avian genotype I in a canary (S. canaria) and an Indian peafowl (Pavo cristatus); Cryptosporidium avian genotype II in ostriches (Struthio camelus) and Cryptosporidium avian genotype III in a cockatiel (N. hollandicus) and a peach-faced lovebird (Agapornis roseicolis).

  4. Phylogenetic Analysis of Cryptosporidium Parasites Based on the Small-Subunit rRNA Gene Locus

    PubMed Central

    Xiao, Lihua; Escalante, Lillian; Yang, Chunfu; Sulaiman, Irshad; Escalante, Anannias A.; Montali, Richard J.; Fayer, Ronald; Lal, Altaf A.

    1999-01-01

    Biological data support the hypothesis that there are multiple species in the genus Cryptosporidium, but a recent analysis of the available genetic data suggested that there is insufficient evidence for species differentiation. In order to resolve the controversy in the taxonomy of this parasite genus, we characterized the small-subunit rRNA genes of Cryptosporidium parvum, Cryptosporidium baileyi, Cryptosporidium muris, and Cryptosporidium serpentis and performed a phylogenetic analysis of the genus Cryptosporidium. Our study revealed that the genus Cryptosporidium contains the phylogenetically distinct species C. parvum, C. muris, C. baileyi, and C. serpentis, which is consistent with the biological characteristics and host specificity data. The Cryptosporidium species formed two clades, with C. parvum and C. baileyi belonging to one clade and C. muris and C. serpentis belonging to the other clade. Within C. parvum, human genotype isolates and guinea pig isolates (known as Cryptosporidium wrairi) each differed from bovine genotype isolates by the nucleotide sequence in four regions. A C. muris isolate from cattle was also different from parasites isolated from a rock hyrax and a Bactrian camel. Minor differences were also detected between C. serpentis isolates from snakes and lizards. Based on the genetic information, a species- and strain-specific PCR-restriction fragment length polymorphism diagnostic tool was developed. PMID:10103253

  5. Molecular characterization of a Cryptosporidium isolate from a banded mongoose Mungos mungo.

    PubMed

    Abe, Niichiro; Takami, Kazutoshi; Kimata, Isao; Iseki, Motohiro

    2004-02-01

    Cryptosporidium spp. has been found in more than 150 species of mammals, but there has been no report in mongooses. In this study, we report the isolation of Cryptosporidium sp. in a banded mongoose Mungos mungo, which was brought from Tanzania to Japan; the isolate was analyzed genetically to validate the occurrence of a new, host-adapted genotype. Cryptosporidium diagnostic fragments of 18S ribosomal RNA and 70-kDa heat shock protein genes were amplified from this isolate and compared with the other Cryptosporidium species and genotypes reported previously. Analyses showed that the mongoose isolate represents a new genotype, closely related to that of bears.

  6. Assessing the infection risk of Giardia and Cryptosporidium in public drinking water delivered by surface water systems in Sao Paulo State, Brazil.

    PubMed

    Sato, Maria Ines Z; Galvani, Ana Tereza; Padula, Jose Antonio; Nardocci, Adelaide Cassia; Lauretto, Marcelo de Souza; Razzolini, Maria Tereza Pepe; Hachich, Elayse Maria

    2013-01-01

    A survey of Giardia and Cryptosporidium was conducted in surface water used as drinking water sources by public water systems in four densely urbanized regions of Sao Paulo State, Brazil. A Quantitative Microbial Risk Assessment, based on protozoa concentrations, was performed to estimate the probability of protozoa infection associated with drinking water ingestion. A total of 206 source water samples were analyzed over a 24 month period using the USEPA Method 1623. The risk of infection was estimated using an exponential dose response model, children and adults exposure and a gamma distribution for (oo)cyst concentrations with three scenarios for treating censored data. Giardia was detected in 102 of the samples, and 19 of them were also positive for Cryptosporidium, with maximum concentrations of 97.0 cysts/L and 6.0 oocysts/L, respectively. Risk distributions were similar for the three scenarios. In the four regions, the estimated risk of Giardia infection per year, for adults and children, ranged from 0.29% to 2.47% and from 0.08% to 0.70%, respectively. Cryptosporidium risk infection varied from 0.15% to 0.29% for adults and from 0.04% to 0.08% for children. In both cases, the calculated risk surpassed the risk of infection of 10(-4) (1:10,000) defined as tolerable by USEPA for a yearly exposure. The probability of Giardia infection was very close to the rates of acute diarrheic disease for adults (1% to 3%) but lower for children (2% to 7%). The daily consumption of drinking water was an important contributing factor for these differences. The Microbiological Risk Assessment carried out in this study provides an indication of infection risks by Giardia and Cryptosporidium in the population served by these source waters. Strategies for source water protection and performance targets for the water treatment should be established to achieve the required level of public health risk.

  7. Utilisation of Rep-PCR to track microbes in aerosols collected adjacent to their source, a saline lake in Victoria, Australia.

    PubMed

    Munday, Chris I; O'Loingsigh, Tadhg; Tapper, Nigel J; De Deckker, Patrick; Allison, Gwen E

    2013-04-15

    Dust storms are a major source of aerosolized bacteria, especially in the drought conditions experienced in Australia in the decade to 2009. The major aims of this project were to identify the culturable bacteria in environmental samples and to genetically fingerprint all isolates using repetitive element PCR (Rep-PCR) to investigate the possibility of tracking isolates from their source into the atmosphere. Four field trips were conducted to a dry lake in western Victoria, Australia to sample aerosols and sediments. Aerosols were collected at heights up to 150 m using vacuum pumps with filters attached to a tethered helium balloon, while corresponding sediments were collected in sterile polypropylene tubes. Isolates were cultivated on Tryptic Soy Agar, R2 Agar and Marine Agar, and grown in dark conditions at ambient temperature. By sequencing the 16S rRNA gene of 270 isolates, fifteen different bacterial families were identified, with both the aerosols and sediments dominated by the Bacillaceae family. Four sets of Rep-PCR primers were tested, with the ERIC and (GTG)5 primers proving to be the most suitable for fingerprinting the cultured taxa. Rep-PCR revealed very high strain diversity in the samples collected, however some strains were still able to be tracked from sediments up to 150 m in height. This shows the potential of Rep-PCR, however very large reference databases would be required for the technique to be more useful.

  8. Preventing community-wide transmission of Cryptosporidium: a proactive public health response to a swimming pool-associated outbreak--Auglaize County, Ohio, USA.

    PubMed

    Cope, J R; Prosser, A; Nowicki, S; Roberts, M W; Roberts, J M; Scheer, D; Anderson, C; Longsworth, A; Parsons, C; Goldschmidt, D; Johnston, S; Bishop, H; Xiao, L; Hill, V; Beach, M; Hlavsa, M C

    2015-12-01

    The incidence of recreational water-associated outbreaks in the United States has significantly increased, driven, at least in part, by outbreaks both caused by Cryptosporidium and associated with treated recreational water venues. Because of the parasite's extreme chlorine tolerance, transmission can occur even in well-maintained treated recreational water venues (e.g. pools) and a focal cryptosporidiosis outbreak can evolve into a community-wide outbreak associated with multiple recreational water venues and settings (e.g. childcare facilities). In August 2004 in Auglaize County, Ohio, multiple cryptosporidiosis cases were identified and anecdotally linked to pool A. Within 5 days of the first case being reported, pool A was hyperchlorinated to achieve 99·9% Cryptosporidium inactivition. A case-control study was launched to epidemiologically ascertain the outbreak source 11 days later. A total of 150 confirmed and probable cases were identified; the temporal distribution of illness onset was peaked, indicating a point-source exposure. Cryptosporidiosis was significantly associated with swimming in pool A (matched odds ratio 121·7, 95% confidence interval 27·4-∞) but not with another venue or setting. The findings of this investigation suggest that proactive implementation of control measures, when increased Cryptosporidium transmission is detected but before an outbreak source is epidemiologically ascertained, might prevent a focal cryptosporidiosis outbreak from evolving into a community-wide outbreak.

  9. Preventing Community-wide Transmission of Cryptosporidium: A Proactive Public Health Response to a Swimming Pool–Associated Outbreak — Auglaize County, Ohio, USA

    PubMed Central

    Cope, J.R.; Prosser, A.; Nowicki, S.; Roberts, M.W.; Scheer, D.; Anderson, C.; Longsworth, A.; Parsons, C.; Goldschmidt, D.; Johnston, S.; Bishop, H.; Xiao, L.; Hill, V.; Beach, M.; Hlavsa, M.C.

    2015-01-01

    Summary The incidence of recreational water–associated outbreaks in the United States has significantly increased, driven, at least in part, by outbreaks both caused by Cryptosporidium and associated with treated recreational water venues. Because of the parasite's extreme chlorine tolerance, transmission can occur even in well-maintained treated recreational water venues, (e.g., pools) and a focal cryptosporidiosis outbreak can evolve into a community-wide outbreak associated with multiple recreational water venues and settings (e.g., child care facilities). In August 2004 in Auglaize County, Ohio, multiple cryptosporidiosis cases were identified and anecdotally linked to Pool A. Within 5 days of the first case being reported, Pool A was hyperchlorinated to achieve 99.9% Cryptosporidium inactivition. A case-control study was launched to epidemiologically ascertain the outbreak source 11 days later. A total of 150 confirmed and probable cases were identified; the temporal distribution of illness onset was peaked, indicating a point-source exposure. Cryptosporidiosis was significantly associated with swimming in Pool A (matched odds ratio 121.7, 95% confidence interval 27.4–∞) but not with another venue or setting. The findings of this investigation suggest that proactive implementation of control measures, when increased Cryptosporidium transmission is detected but before an outbreak source is epidemiologically ascertained, might prevent a focal cryptosporidiosis outbreak from evolving into a community-wide outbreak. PMID:25907106

  10. Characterization of Cryptosporidium spp.--recent developments and future needs.

    PubMed

    Egyed, Z; Sréter, T; Széll, Z; Varga, I

    2003-02-13

    Cryptosporidia, widely distributed protozoan parasites of vertebrates have recently attracted increasing interest due to several serious waterborne outbreaks, the life-threatening nature of infection in immunocompromised patients, and the realization of economic losses caused by these pathogens in livestock. Genetic polymorphism within Cryptosporidium spp. is being detected at a continuously growing rate, owing to the widespread use of modern molecular techniques. The aim of this paper is to review the current status of taxonomy, genotyping, molecular phylogeny, and characterization of cryptosporidia, and to highlight the need for polyphasic typing, i.e. an integrated approach comprising standardized morphologic, biologic, and molecular methods for describing Cryptosporidium species and isolates, and for establishing "virtual" reference strains.

  11. Effect of water treatment processes on Cryptosporidium infectivity.

    PubMed

    Keegan, Alexandra; Daminato, David; Saint, Christopher P; Monis, Paul T

    2008-03-01

    Conventional water treatment processes have the ability to remove Cryptosporidium oocysts through coagulation, flocculation, sedimentation and filtration, provided there is efficient management of plant performance. The potential exists for the breakthrough of oocysts through the treatment train. The effect of the water treatment chemical aluminium sulphate (alum) on Cryptosporidium oocyst infectivity has been assessed using an assay that combines cell culture and real-time polymerase chain reaction techniques. The infectivity of fresh and temperature-aged oocysts (stored up to 6 months at 4 or 15 degrees C) was unaffected by exposure to a range of doses of alum in standard jar test procedures and dissolved air flotation processes and subsequent exposure to chlorine or chloramine. Removal efficiencies and infectivity measures are important in determining risk to public health and will reflect the ability of water treatment plants to act as a barrier to these pathogens.

  12. Paired city cryptosporidium serosurvey in the southwest USA.

    PubMed Central

    Frost, F. J.; Muller, T.; Craun, G. F.; Calderon, R. L.; Roefer, P. A.

    2001-01-01

    In 1996, serological responses to two Cryptosporidium antigens were determined for 200 Las Vegas (LV), Nevada, and 200 Albuquerque, New Mexico, blood donors to evaluate associations between endemic infections, water exposures, and other risk factors. LV uses chlorinated filtered drinking water from Lake Mead while Albuquerque uses chlorinated ground water. The intensity of serological response to both markers was higher for older donors (P < 0.05). donors who washed food with bottled water (P < 0.05) and donors from LV (P < 0.05). A decreased serological response was not associated with bottled water consumption, nor was an increased response associated with self-reported cryptosporidiosis-like illness or residence in LV at the time of a cryptosporidiosis outbreak 2 years earlier. Although these findings suggest the serological response may be associated with type of tap water and certain foods, additional research is needed to clarifythe role of both food and drinking water in endemic Cryptosporidium infection. PMID:11349981

  13. Occurrence of Giardia and Cryptosporidium in pigs on Prince Edward Island, Canada.

    PubMed

    Budu-Amoako, Ebo; Greenwood, Spencer J; Dixon, Brent R; Barkema, Herman W; Hurnik, Daniel; Estey, Chelsie; McClure, J T

    2012-02-28

    In a cross-sectional study of 633 pigs from 21 herds on Prince Edward Island, Canada (PEI), the prevalence of infection with Cryptosporidium and Giardia, and the genotypes and species of isolates were determined in order to establish the zoonotic potential of pigs in this region. As determined by direct immunofluorescence microscopy (DFA), 18 herds (86%) and 163 animals (26%; 95% CI: 22-29%) tested positive for Cryptosporidium, while just 3 herds (14%) and 6 animals (1%; 95% CI: 0.4-2%) tested positive for Giardia. Cryptosporidium spp. isolates were detected in 39% (95% CI: 34-44%) of weanlings (1-3 months of age) and 9% (95% CI: 6-13) of sows (>8months of age). Molecular characterization using the 18S rDNA and HSP70 gene fragments revealed the presence of Cryptosporidium sp. pig genotype II, C. suis, C. parvum, and Cryptosporidium sp. mouse genotype. Among the 113 isolates of Cryptosporidium spp. successfully genotyped, pig genotype II (61%) predominated, with C. suis (36%) being the next most prominant isolate. C. parvum (2%; two isolates) and Cryptosporidium sp. mouse genotype (0.9%; one isolate) were only occasionally isolated. The only two Cryptosporidium-positive genotyped isolates from sows included one each of C. suis and Cryptosporidium sp. pig genotype II. All but one of the six Giardia positive isolates were detected in weanling pigs. None of the Giardia-positive isolates was amenable to PCR. This study demonstrates that Cryptosporidium spp. are highly prevalent in pigs on PEI, Canada, are found mostly in weanlings (1-3 months of age). Furthermore, the pigs are primarily infected by the host-specific genotypes and species, Cryptosporidium sp. pig genotype II and C. suis, whereas the zoonotic C. parvum is rare. Giardia duodenalis is only occasionally found in pigs. These findings suggest that domestic pigs on PEI, Canada, likely do not pose a significant health risk to humans from these parasites.

  14. Molecular Characterization of Isolates of Waterborne Cryptosporidium spp. Collected during an Outbreak of Gastroenteritis in South Burgundy, France

    PubMed Central

    Dalle, Frédéric; Roz, Pascale; Dautin, Guillaume; Di-Palma, Marc; Kohli, Evelyne; Sire-Bidault, C.; Fleischmann, Marie George; Gallay, Anne; Carbonel, Sylvia; Bon, Fabienne; Tillier, Claude; Beaudeau, Pascal; Bonnin, Alain

    2003-01-01

    In September 2001, a waterborne outbreak of gastroenteritis occurred in eastern France. Of 31 fecal samples from symptomatic individuals, 19 tested positive for Cryptosporidium with two PCRs targeting the Hsp70 and the 18S rRNA genes of Cryptosporidium. Sequencing of the PCR fragments produced sequences identical to that of Cryptosporidium parvum genotype 1. PMID:12791906

  15. Molecular characterization of isolates of waterborne Cryptosporidium spp. collected during an outbreak of gastroenteritis in South Burgundy, France.

    PubMed

    Dalle, Frédéric; Roz, Pascale; Dautin, Guillaume; Di-Palma, Marc; Kohli, Evelyne; Sire-Bidault, C; Fleischmann, Marie George; Gallay, Anne; Carbonel, Sylvia; Bon, Fabienne; Tillier, Claude; Beaudeau, Pascal; Bonnin, Alain

    2003-06-01

    In September 2001, a waterborne outbreak of gastroenteritis occurred in eastern France. Of 31 fecal samples from symptomatic individuals, 19 tested positive for Cryptosporidium with two PCRs targeting the Hsp70 and the 18S rRNA genes of CRYPTOSPORIDIUM: Sequencing of the PCR fragments produced sequences identical to that of Cryptosporidium parvum genotype 1.

  16. Molecular Surveillance of Cryptosporidium spp. in Raw Wastewater in Milwaukee: Implications for Understanding Outbreak Occurrence and Transmission Dynamics

    PubMed Central

    Zhou, Ling; Singh, Ajaib; Jiang, Jianlin; Xiao, Lihua

    2003-01-01

    Six Cryptosporidium spp. were found in 50 of 179 Milwaukee wastewater samples collected weekly over a year. Of the eight subtypes of Cryptosporidium hominis and Cryptosporidium parvum present, allele Ib was found in 14 of 16 samples, and its sequence was identical to that of the subtype in human samples from the 1993 Milwaukee outbreak of cryptosporidiosis. PMID:14605176

  17. Detection and Characterization of a Cryptosporidium Isolate from a Southern Elephant Seal (Mirounga leonina) from the Antarctic Peninsula ▿

    PubMed Central

    Rengifo-Herrera, C.; Ortega-Mora, L. M.; Gómez-Bautista, M.; García-Moreno, F. T.; García-Párraga, D.; Castro-Urda, J.; Pedraza-Díaz, S.

    2011-01-01

    The presence of Cryptosporidium and Giardia in 221 fecal samples from different species of Antarctic pinnipeds was investigated by immunofluorescence microscopy and PCR. Cryptosporidium, a skunk-like genotype, was detected only in a southern elephant seal. Giardia was not detected. This is the first report of a Cryptosporidium sp. in Antarctic marine mammals. PMID:21169427

  18. Infectious Cryptosporidium parvum oocysts in final reclaimed effluent

    USGS Publications Warehouse

    Gennaccaro, A.L.; McLaughlin, M.R.; Quintero-Betancourt, W.; Huffman, D.E.; Rose, J.B.

    2003-01-01

    Water samples collected throughout several reclamation facilities were analyzed for the presence of infectious Cryptosporidium parvum by the focus detection method-most-probable-number cell culture technique. Results revealed the presence of infectious C. parvum oocysts in 40% of the final disinfected effluent samples. Sampled effluent contained on average seven infectious oocysts per 100 liters. Thus, reclaimed water is not pathogen free but contains infectious C. parvum.

  19. Cryptosporidium Infection Risk: Results of New Dose-Response Modeling.

    PubMed

    Messner, Michael J; Berger, Philip

    2016-10-01

    Cryptosporidium human dose-response data from seven species/isolates are used to investigate six models of varying complexity that estimate infection probability as a function of dose. Previous models attempt to explicitly account for virulence differences among C. parvum isolates, using three or six species/isolates. Four (two new) models assume species/isolate differences are insignificant and three of these (all but exponential) allow for variable human susceptibility. These three human-focused models (fractional Poisson, exponential with immunity and beta-Poisson) are relatively simple yet fit the data significantly better than the more complex isolate-focused models. Among these three, the one-parameter fractional Poisson model is the simplest but assumes that all Cryptosporidium oocysts used in the studies were capable of initiating infection. The exponential with immunity model does not require such an assumption and includes the fractional Poisson as a special case. The fractional Poisson model is an upper bound of the exponential with immunity model and applies when all oocysts are capable of initiating infection. The beta Poisson model does not allow an immune human subpopulation; thus infection probability approaches 100% as dose becomes huge. All three of these models predict significantly (>10x) greater risk at the low doses that consumers might receive if exposed through drinking water or other environmental exposure (e.g., 72% vs. 4% infection probability for a one oocyst dose) than previously predicted. This new insight into Cryptosporidium risk suggests additional inactivation and removal via treatment may be needed to meet any specified risk target, such as a suggested 10(-4) annual risk of Cryptosporidium infection.

  20. Intestinal immune response to human Cryptosporidium sp. infection

    DTIC Science & Technology

    2008-01-01

    and eliminate the infection, which typically causes acute, self-limited watery diarrhea lasting 5 to 10 days. However, in patients with defects in ...cellular immune responses (e.g., AIDS, malnutrition, or defects in the CD40- CD154 system), Cryptosporidium frequently causes persistent or chronic...diarrhea and may also involve the biliary tract (40). In malnourished children, persistent diarrhea is associated with increased susceptibility to recurrent

  1. Efficacy of chlorine dioxide tablets on inactivation of cryptosporidium oocysts.

    PubMed

    Murphy, Jennifer L; Haas, Charles N; Arrowood, Michael J; Hlavsa, Michele C; Beach, Michael J; Hill, Vincent R

    2014-05-20

    The ability of chlorine dioxide (ClO2) to achieve 2-log inactivation of Cryptosporidium in drinking water has been documented. No studies have specifically addressed the effects of ClO2 on C. parvum oocyst infectivity in chlorinated recreational water venues (e.g., pools). The aim of this research was to determine the efficacy of ClO2 as an alternative to existing hyperchlorination protocols that are used to achieve a 3-log inactivation of Cryptosporidium in such venues. To obtain a 3-log inactivation of C. parvum Iowa oocysts, contact times of 105 and 128 min for a solution containing 5 mg/L ClO2 with and without the addition of 2.6 mg/L free chlorine, respectively, were required. Contact times of 294 and 857 min for a solution containing 1.4 mg/L ClO2 with and without the addition of 3.6 mg/L free chlorine, respectively, were required. The hyperchlorination control (21 mg/L free chlorine only) required 455 min for a 3-log inactivation. Use of a solution containing 5 mg/L ClO2 and solutions containing 5 or 1.4 mg/L ClO2 with the addition of free chlorine appears to be a promising alternative to hyperchlorination for inactivating Cryptosporidium in chlorinated recreational water venues, but further studies are required to evaluate safety constraints on use.

  2. Morphological and immunohistochemical features of Cryptosporidium andersoni in cattle.

    PubMed

    Masuno, K; Yanai, T; Hirata, A; Yonemaru, K; Sakai, H; Satoh, M; Masegi, T; Nakai, Y

    2006-03-01

    Light and electron microscopic features and immunohistochemical features of Cryptosporidium andersoni (C. andersoni) and host reaction in the mucosa were studied. Although the affected cattle demonstrated no apparent clinical signs, a severe infection of C. andersoni was observed in the abomasum. C. andersoni were round in shape, measured 6-8 microm in size and were mainly observed to be freely located in the gastric pits, being attached in occasional cases to the surface of the abomasum epithelium. Frequent inflammatory cells had infiltrated the lamina propria of the affected mucosa, and frequent mitotic figures were observed in epithelial cells at the dilated isthmus. To access the cell kinetics, the number of epithelial cells infected with C. andersoni were counted and compared with noninfected cattle. The number of gastric pit cells in infected cattle was significantly higher than that in the controls. The number of proliferative cells determined by the Ki-67 antigen in C. andersoni infected cattle was also significantly higher than that in the controls. Transmission electron microscopy and scanning electron microscopy revealed that the morphology of the C. andersoni organism was common to those of other Cryptosporidium spp. Immunohistochemically, several commercial antibodies against Cryptosporidium spp. showed positive reactions at the wall of these oocysts or parasitophorous vacuoles. This report is possibly the first to discuss the prominent hyperplasia of the abomasum mucosa, as well as morphologic features of C. andersoni in cattle.

  3. Molecular Characterization of Cryptosporidium spp. in Children from Mexico

    PubMed Central

    Valenzuela, Olivia; González-Díaz, Mariana; Garibay-Escobar, Adriana; Burgara-Estrella, Alexel; Cano, Manuel; Durazo, María; Bernal, Rosa M.; Hernandez, Jesús; Xiao, Lihua

    2014-01-01

    Cryptosporidiosis is a parasitic disease caused by Cryptosporidium spp. In immunocompetent individuals, it usually causes an acute and self-limited diarrhea; in infants, infection with Cryptosporidium spp. can cause malnutrition and growth retardation, and declined cognitive ability. In this study, we described for the first time the distribution of C. parvum and C. hominis subtypes in 12 children in Mexico by sequence characterization of the 60-kDa glycoprotein (GP60) gene of Cryptosporidium. Altogether, 7 subtypes belonging to 4 subtype families of C. hominis (Ia, Ib, Id and Ie) and 1 subtype family of C. parvum (IIa) were detected, including IaA14R3, IaA15R3, IbA10G2, IdA17, IeA11G3T3, IIaA15G2R1 and IIaA16G1R1. The frequency of the subtype families and subtypes in the samples analyzed in this study differed from what was observed in other countries. PMID:24755606

  4. Cryptosporidium parvum DNA replication in cell-free culture.

    PubMed

    Zhang, L; Sheoran, A S; Widmer, G

    2009-10-01

    The lack of robust methods for culturing Cryptosporidium parasites remains a major challenge and is hampering efforts to screen for anti-cryptosporidial drugs. In existing culture methods, monolayers of mammalian epithelial cells are inoculated with oocysts. The system supports an initial phase of asexual proliferation of the parasite. For reasons that are not clear, development rapidly declines within 2-3 days. The unexpected report of Cryptosporidium parvum culture in the absence of host cells, and the failure of others to reproduce the method, prompted us to apply quantitative PCR to measure changes in C. parvum DNA levels in cell-free cultures, and parasite-specific antibodies to identify different life cycle stages. Based on this approach, which has not been applied previously to analyze C. parvum growth in cell-free culture, we found that the concentration of C. parvum DNA increased by about 5-fold over 5 days of culture. Immuno-labeling of cultured organisms revealed morphologically distinct stages, only some of which reacted with Cryptosporidium-specific monoclonal antibodies. These observations are indicative of a modest proliferation of C. parvum in cell-free culture.

  5. Molecular characterization of Cryptosporidium spp. in children from Mexico.

    PubMed

    Valenzuela, Olivia; González-Díaz, Mariana; Garibay-Escobar, Adriana; Burgara-Estrella, Alexel; Cano, Manuel; Durazo, María; Bernal, Rosa M; Hernandez, Jesús; Xiao, Lihua

    2014-01-01

    Cryptosporidiosis is a parasitic disease caused by Cryptosporidium spp. In immunocompetent individuals, it usually causes an acute and self-limited diarrhea; in infants, infection with Cryptosporidium spp. can cause malnutrition and growth retardation, and declined cognitive ability. In this study, we described for the first time the distribution of C. parvum and C. hominis subtypes in 12 children in Mexico by sequence characterization of the 60-kDa glycoprotein (GP60) gene of Cryptosporidium. Altogether, 7 subtypes belonging to 4 subtype families of C. hominis (Ia, Ib, Id and Ie) and 1 subtype family of C. parvum (IIa) were detected, including IaA14R3, IaA15R3, IbA10G2, IdA17, IeA11G3T3, IIaA15G2R1 and IIaA16G1R1. The frequency of the subtype families and subtypes in the samples analyzed in this study differed from what was observed in other countries.

  6. Hydrophobic and electrostatic cell surface properties of Cryptosporidium parvum.

    PubMed

    Drozd, C; Schwartzbrod, J

    1996-04-01

    Microbial adhesion to hydrocarbons and microelectrophoresis were investigated in order to characterize the surface properties of Cryptosporidium parvum. Oocysts exhibited low removal rates by octane (only 20% on average), suggesting that the Cryptosporidium sp. does not demonstrate marked hydrophobic properties. A zeta potential close to -25 mV at pH 6 to 6.5 in deionized water was observed for the parasite. Measurements of hydrophobicity and zeta potential were performed as a function of pH and ionic strength or conductivity. Hydrophobicity maxima were observed at extreme pH values, with 40% of adhesion of oocysts to octane. It also appeared that ionic strength (estimated by conductivity) could influence the hydrophobic properties of oocysts. Cryptosporidium oocysts showed a pH-dependent surface charge, with zeta potentials becoming less negative as pH was reduced, starting at -35 mV for alkaline pH and reaching 0 at isoelectric points for pH 2.5. On the other hand, variation of surface charge with respect to conductivity of the suspension tested in this work was quite small. The knowledge of hydrophobic properties and surface charge of the parasite provides information useful in, for example, the choice of various flocculation treatments, membrane filters, and cleaning agents in connection with oocyst recovery.

  7. Molecular epidemiological analyses of Cryptosporidium parvum virus 1 (CSpV1), a symbiotic virus of Cryptosporidium parvum, in Japan.

    PubMed

    Murakoshi, Fumi; Ichikawa-Seki, Madoka; Aita, Junya; Yaita, Seiko; Kinami, Aiko; Fujimoto, Katsuhisa; Nishikawa, Yoshifumi; Murakami, Shin; Horimoto, Taisuke; Kato, Kentaro

    2016-01-04

    We show that Cryptosporidium parvum virus 1 (CSpV1), a member of the family Partitiviridae, genus Cryspovirus that can infect Cryptosporidium parvum, is a new candidate for high-resolution tool for tracing C. parvum. CSpV1 was detected in all C. parvum-positive samples tested. Phylogenetic analysis of dsRNA1 sequence from CSpV1 can distinguish infected areas of C. parvum on the national level. Sequences detected in samples from Iwate prefecture and other islands (Tanegashima, and Okinawa) belonged to a single clade. This system can differentiate the samples from Hokkaido and south part of Japan as well as from other countries. Samples from Iwate, Tanegashima, and Okinawa belonged to a single subclade, respectively. Therefore, the CSpV1 dsRNA sequences reflect the regional distribution of their host and have potential as a high-resolution tool to trace C. parvum IIaA15G2R1 subtype.

  8. A dual-plane co-RASOR technique for accurate and rapid tracking and position verification of an Ir-192 source for single fraction HDR brachytherapy

    NASA Astrophysics Data System (ADS)

    de Leeuw, Hendrik; Moerland, Marinus A.; van Vulpen, Marco; Seevinck, Peter R.; Bakker, Chris J. G.

    2013-11-01

    Effective high-dose-rate (HDR) treatment requires accurate and independent treatment verification to ensure that the treatment proceeds as prescribed, in particular if a high dose is given, as in single fraction therapy. Contrary to CT imaging and fluoroscopy, MR imaging provides high soft tissue contrast. Conventional MR techniques, however, do not offer the temporal resolution in combination with the 3D spatial resolution required for accurate brachytherapy source localization. We have developed an MR imaging method (center-out RAdial Sampling with Off-Resonance (co-RASOR)) that generates high positive contrast in the geometrical center of field perturbing objects, such as HDR brachytherapy sources. co-RASOR generates high positive contrast in the geometric center of an Ir-192 source by applying a frequency offset to center-out encoded data. To obtain high spatial accuracy in 3D with adequate temporal resolution, two orthogonal center-out encoded 2D images are applied instead of a full 3D acquisition. Its accuracy in 3D is demonstrated by 3D MRI and CT. The 2D images show high positive contrast in the geometric center of non-radioactive Ir-192 sources, with signal intensities up to 160% of the average signal intensity in the surrounding medium. The accuracy with which the center of the Ir-192 source is located by the dual-plane MRI acquisition corresponds closely to the accuracy obtained by 3D MRI and CT imaging. The positive contrast is shown to be obtained in homogeneous and in heterogeneous tissue. The dual-plane MRI technique allows the brachytherapy source to be tracked in 3D with millimeter accuracy with a temporal resolution of approximately 4 s.

  9. Cryptosporidium Off-The-Slide Genotyping: Recovery of Oocysts Extracted from Slides Examined by Method 1623

    EPA Science Inventory

    Cryptosporidium has been found world wide and is an important waterborne parasite causing a self-limiting diarrhea; however, in the immunocompromised it may become chronic and can even lead to death. To characterize the risk of exposure to Cryptosporidium in water the USEPA enac...

  10. INVESTIGATION AND EVALUATION OF CRYPTOSPORIDIUM AND GIARDIA IN COMBINED SEWER OVERFLOW (CSO) AND STORMWATER RUNOFF

    EPA Science Inventory

    Since the first identified Cryptosporidium outbreaks occurred in the 1980s and the massive 1993 Milwaukee, WI outbreak affected more than 400,000 people, the concern over the public health risks related to protozoan pathogens Cryptosporidium and Giardia has g...

  11. GENOTYPES OF CRYPTOSPORIDIUM INFECTING FUR-BEARING MAMMALS DIFFER FROM THOSE INFECTING HUMANS

    EPA Science Inventory

    To evaluate the public health significance of wildlife Cryptosporidium spp., five species of small wild mammals (beavers, muskrats, otters, raccoons, and foxes) living in Chesapeake Bay watersheds were examined in this study for the presence of Cryptosporidium spp., using a small...

  12. Wide distribution of Cryptosporidium bovis and the deer-like genotype in bovines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We recently reported that on 14 dairy farms from Vermont to Florida ~85% of pre-weaned dairy calves were infected with zoonotic Cryptosporidium parvum whereas only 1-2% of post-weaned calves and 1-2 year-old heifers were infected with this species. Cryptosporidium bovis and the deer-like genotype w...

  13. Spinacia oleracea L. leaf stomata harboring Cryptosporidium parvum oocysts: A potential threat for food safety

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Scientific literature documents the prevalence of Cryptosporidium oocysts in irrigation waters and on fresh produce. In the present study spinach leaves were experimentally exposed to Cryptosporidium oocysts which were subsequently irrigated with clean water daily for 5 days. As determined by confoc...

  14. CRYPTOSPORIDIUM AND GIARDIA IN STORMWATER AS A THREAT TO DRINKING WATER SUPPLIES

    EPA Science Inventory

    Since the first identified Cryptosporidium outbreak in the United Kingdom in 1983, the pathogens Cryptosporidium and Giardia have become the subject of growing local, state, and national concern. In the last decade, these organisms have been the causative agent of many gastroint...

  15. EPA ORD/REGION 2 COOPERATIVE EFFORT: EVALUATING CRYPTOSPORIDIUM AND GIARDIA IN URBAN STORMWATER

    EPA Science Inventory

    Since the first identified Cryptosporidium outbreak in the United Kingdom in 1983, the pathogens Cryptosporidium and Giardia have become the subject of growing local, state, and national concern. Both organisms have been the causative agent of many gas...

  16. EVALUATING CRYPTOSPORIDIUM AND GIARDIA IN STORMWATER AS A THREAT TO DRINKING WATER SUPPLIES

    EPA Science Inventory

    Since the first identified Cryptosporidium outbreak in the United Kingdom in 1983, the
    pathogens Cryptosporidium and Giardia have become the subject of growing local, state, and
    national concern. Both organisms have been the causative agent of many gastrointestinal
    illn...

  17. Cryptosporidium muris, a Rodent Pathogen, Recovered from a Human in Perú

    PubMed Central

    Xiao, Lihua; Terashima, Angélica; Guerra, Humberto; Gotuzzo, Eduardo; Saldías, Gustavo; Bonilla, J. Alfredo; Zhou, Ling; Lindquist, Alan; Upton, Steve J.

    2003-01-01

    Cryptosporidium muris, predominantly a rodent species of Cryptosporidium, is not normally considered a human pathogen. Recently, isolated human infections have been reported from Indonesia, Thailand, France, and Kenya. We report the first case of C. muris in a human in the Western Hemisphere. This species may be an emerging zoonotic pathogen capable of infecting humans. PMID:14519260

  18. Prevalence rates of Giardia and Cryptosporidium among diarrheic patients in the Philippines.

    PubMed

    Natividad, Filipinas F; Buerano, Corazon C; Lago, Catherine B; Mapua, Cynthia A; de Guzman, Blanquita B; Seraspe, Ebonia B; Samentar, Lorena P; Endo, Takuro

    2008-11-01

    The prevalence of Giardia and Cryptosporidium among 3,456 diarrheic patients corrected from May 2004 to May 2005 in the Philippines was determined. Of 133 (3.8%) positive samples, 69 (2.0%) were positive for Giardia and 67 (1.9%) for Cryptosporidium. Three samples had co-infection with Giardia and Cryptosporidium. Luzon had the highest positive samples (5.0%) followed by Mindanao (4.9%), then Visayas (2.2%). Giardia was most prevalent in Mindanao (3.6%) while Cryptosporidium was most prevalent in Luzon (3.1%). The prevalence of Giardia (2.0%) among pediatric patients (0-18 years) did not significantly differ from that (1.9%) among adults (> 18 years old). However, for Cryptosporidium, the prevalence (2.9%) among pediatric patients was significantly higher compared to that (0.2%) among adult patients. In the pediatric population, the highest percentage of patients with Giardia was the 5-9 year old age group, while that of Cryptosporidium was in the 0-4 year old group. The prevalence of Giardia, but not Cryptosporidium, was significantly higher in male than female adults. Seasonality had a distinct peak in September with Cryptosporidium more prevalent in the rainy (2.6%) than dry season (0.9%).

  19. Cryptosporidium from a free-ranging marsupial host: bandicoots in urban Australia.

    PubMed

    Dowle, Matthew; Hill, Nichola J; Power, Michelle L

    2013-11-15

    Expansion of human settlement has increased the interface between people and bandicoots with implications for the emergence and spread of zoonotic parasites. The host status of bandicoots inhabiting suburban areas and their potential role in Cryptosporidium transmission remains unresolved. Our study aimed to determine the prevalence and identity of Cryptosporidium in two sympatric bandicoot species. Cryptosporidium signatures were detected in twelve bandicoot faecal samples (n=98) through amplification of the 18S rRNA. Phylogenetic inference placed the isolates in a clade with Cryptosporidium parvum, a species with a broad host range and zoonotic potential, or loosely related to Cryptosporidium hominis. However, the identity of the bandicoot isolates was not fully resolved and whether they were infected or simply passively transmitting oocysts is unknown. This study revealed that free-ranging bandicoots of northern Sydney were shedding Cryptosporidium oocysts at a prevalence of 12.2% (95% CI [6.76, 20.8]), similar to marsupial species that act as reservoirs for Cryptosporidium. Our findings expand the range of hosts known to shed Cryptosporidium in urban areas.

  20. Cryptosporidium oocysts and giardia cysts on salad products irrigated with contaminated water.

    PubMed

    Amorós, Inmaculada; Alonso, José L; Cuesta, Gonzalo

    2010-06-01

    A field study in Valencia, Spain, was done to determine the occurrence of Giardia and Cryptosporidium on salad products that are frequently eaten raw, such as lettuces and Chinese cabbage, and in irrigation waters. Four water samples were taken weekly 1 month before harvesting the vegetables. All water samples were analyzed using techniques included in the U.S. Environmental Protection Agency Method 1623. Standard methods for detecting protozoan parasites on salad vegetables are not available. Published techniques for the isolation of parasites from vegetables generally have low and variable recovery efficiencies. In this study, vegetables were analyzed using a recently reported method for detection of Cryptosporidium oocysts and Giardia cysts on salad products. The waters tested were positive for both Cryptosporidium and Giardia. Of 19 salad products studied, we observed Cryptosporidium in 12 samples and Giardia in 10 samples. Recoveries of the Texas Red-stained Cryptosporidium and Giardia, which were used as internal controls, were 24.5% +/- 3.5% for Cryptosporidium and 16.7% +/- 8.1% for Giardia (n = 8). This study provides data on the occurrence of Cryptosporidium and Giardia in salad products in Spain. The method was useful in the detection of Cryptosporidium oocysts and Giardia cysts on the vegetables tested, and it provides a useful analytical tool for occurrence monitoring.

  1. The first large scale molecular study of Cryptosporidium and Giardia in horses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The prevalence of species and genotypes of Giardia and Cryptosporidium in horses is poorly known. The present study examined feces from 195 horses, 1 month to 17 years of age, in 4 locations in Colombia. Prevalence and age distribution of Giardia and Cryptosporidium were determined by PCR. All PCR p...

  2. INVESTIGATION AND EVALUATION OF CRYPTOSPORIDIUM AND GIARDIA IN COMBINED SEWER OVERFLOW (CSO) AND STORMWATER RUNOFF

    EPA Science Inventory

    Since the first identified Cryptosporidium outbreaks occurred in the 1980s and the massive 1993 Milwaukee, Wisconsin outbreak affected more than 400,000 people (Fox & Lytle 1996), the concern over the public health risks related to protozoan pathogens Cryptosporidium

  3. Tracking the Primary Sources of Fecal Pollution in a Tropical Watershed in a One-Year Study

    EPA Science Inventory

    A study was conducted to determine the primary sources of fecal pollution in a subtropical watershed using host-specific assays developed in temperate regions. Water samples (n=534) from 10 different sites along the Rio Grande de Arecibo watershed were collected every two-three w...

  4. Multiplex PCR detection of waterborne intestinal protozoa: microsporidia, Cyclospora, and Cryptosporidium.

    PubMed

    Lee, Seung-Hyun; Joung, Migyo; Yoon, Sejoung; Choi, Kyoungjin; Park, Woo-Yoon; Yu, Jae-Ran

    2010-12-01

    Recently, emerging waterborne protozoa, such as microsporidia, Cyclospora, and Cryptosporidium, have become a challenge to human health worldwide. Rapid, simple, and economical detection methods for these major waterborne protozoa in environmental and clinical samples are necessary to control infection and improve public health. In the present study, we developed a multiplex PCR test that is able to detect all these 3 major waterborne protozoa at the same time. Detection limits of the multiplex PCR method ranged from 10(1) to 10(2) oocysts or spores. The primers for microsporidia or Cryptosporidium used in this study can detect both Enterocytozoon bieneusi and Encephalitozoon intestinalis, or both Cryptosporidium hominis and Cryptosporidium parvum, respectively. Restriction enzyme digestion of PCR products with BsaBI or BsiEI makes it possible to distinguish the 2 species of microsporidia or Cryptosporidium, respectively. This simple, rapid, and cost-effective multiplex PCR method will be useful for detecting outbreaks or sporadic cases of waterborne protozoa infections.

  5. Molecular analysis of Cryptosporidium from cattle from five states of Peninsular Malaysia.

    PubMed

    Yap, Nan Jiun; Koehler, Anson V; Ebner, Janine; Tan, Tiong Kai; Lim, Yvonne A L; Gasser, Robin B

    2016-02-01

    Despite the importance of the cattle industry in Malaysia, there are very few studies of the diversity and public health significance of bovine cryptosporidiosis in this country. In the present study, we used a PCR-based approach to detect and genetically characterize Cryptosporidium DNA in faecal samples from a cohort of 215 asymptomatic cattle (of different ages) from six farms from five states of Peninsular Malaysia. Cattle on four of the six farms were test-positive for Cryptosporidium, with an overall prevalence of 3.2%. Cryptosporidium bovis and Cryptosporidium ryanae were detected in two (0.9%) and five (2.3%) samples tested; this low prevalence likely relates to the age of the cattle tested, as most (73%) of the samples tested originated from cattle that were ≥2 years of age. Future studies should investigate the zoonotic potential of Cryptosporidium in pre-weaned and weaned calves in rural communities of Malaysia.

  6. It's official - Cryptosporidium is a gregarine: What are the implications for the water industry?

    PubMed

    Ryan, Una; Paparini, Andrea; Monis, Paul; Hijjawi, Nawal

    2016-11-15

    Parasites of the genus Cryptosporidium are a major cause of diarrhoea and ill-health in humans and animals and are frequent causes of waterborne outbreaks. Until recently, it was thought that Cryptosporidium was an obligate intracellular parasite that only replicated within a suitable host, and that faecally shed oocysts could survive in the environment but could not multiply. In light of extensive biological and molecular data, including the ability of Cryptosporidium to complete its life cycle in the absence of a host and the production of novel extracellular stages, Cryptosporidium has been formally transferred from the Coccidia, to a new subclass, Cryptogregaria, with gregarine parasites. In this review, we discuss the close relationship between Cryptosporidium and gregarines and discuss the implications for the water industry.

  7. Occurrence of Cryptosporidium and Giardia in raw and finished drinking water in north-eastern Spain.

    PubMed

    Ramo, Ana; Del Cacho, Emilio; Sánchez-Acedo, Caridad; Quílez, Joaquín

    2017-02-15

    This paper collects the first large-sample-size study on the presence of Cryptosporidium oocysts and Giardia cysts in drinking water plants at the 20 most populated towns in Aragón (north-eastern Spain). Samples of influent raw water and effluent finished water were collected from each plant during different seasons and processed according to USEPA Method 1623. Cryptosporidium oocysts and Giardia cysts were detected in samples collected from 55% and 70% plants, respectively, with nine plants being positive for both protozoa and only four plants being negative over the study period. Both parasites were identified in the raw water throughout the year, with a lower frequency in autumn and a peak in winter, at a mean concentration of 67±38 oocysts per 100l and 125±241 cysts per 100l. The turbidity of raw water was not related to the presence or concentration of (oo)cysts, and the (oo)cyst removal efficiency was not related to the type of water treatment. One or both pathogens were identified in the finished water in 7 out of 11 plants with a conventional treatment process (coagulation, flocculation, sedimentation, filtration, and disinfection processes) compared to 4 out of 9 plants that did not apply one of the pre-chlorination treatment steps. Protozoa were detected in the finished water of positive plants at a mean concentration of 88±55 oocysts per 100l and 37±41 cysts per 100l, and most of them excluded propidium iodide so were considered potentially viable. The ubiquity of these parasites in the drinking water sources and the inefficiency of conventional water treatment in reducing/inactivating them may present a serious public health issue in this geographical area.

  8. Chlorine Dioxide Inactivation of Cryptosporidium parvum Oocysts and Bacterial Spore Indicators

    PubMed Central

    Chauret, Christian P.; Radziminski, Chris Z.; Lepuil, Michael; Creason, Robin; Andrews, Robert C.

    2001-01-01

    Cryptosporidium parvum, which is resistant to chlorine concentrations typically used in water treatment, is recognized as a significant waterborne pathogen. Recent studies have demonstrated that chlorine dioxide is a more efficient disinfectant than free chlorine against Cryptosporidium oocysts. It is not known, however, if oocysts from different suppliers are equally sensitive to chlorine dioxide. This study used both a most-probable-number–cell culture infectivity assay and in vitro excystation to evaluate chlorine dioxide inactivation kinetics in laboratory water at pH 8 and 21°C. The two viability methods produced significantly different results (P < 0.05). Products of disinfectant concentration and contact time (Ct values) of 1,000 mg · min/liter were needed to inactivate approximately 0.5 log10 and 2.0 log10 units (99% inactivation) of C. parvum as measured by in vitro excystation and cell infectivity, respectively, suggesting that excystation is not an adequate viability assay. Purified oocysts originating from three different suppliers were evaluated and showed marked differences with respect to their resistance to inactivation when using chlorine dioxide. Ct values of 75, 550, and 1,000 mg · min/liter were required to achieve approximately 2.0 log10 units of inactivation with oocysts from different sources. Finally, the study compared the relationship between easily measured indicators, including Bacillus subtilis (aerobic) spores and Clostridium sporogenes (anaerobic) spores, and C. parvum oocysts. The bacterial spores were found to be more sensitive to chlorine dioxide than C. parvum oocysts and therefore could not be used as direct indicators of C. parvum inactivation for this disinfectant. In conclusion, it is suggested that future studies address issues such as oocyst purification protocols and the genetic diversity of C. parvum, since these factors might affect oocyst disinfection sensitivity. PMID:11425712

  9. Can oxygen stable isotopes be used to track precipitation moisture source in vascular plant-dominated peatlands?

    NASA Astrophysics Data System (ADS)

    Amesbury, Matthew J.; Charman, Dan J.; Newnham, Rewi M.; Loader, Neil J.; Goodrich, Jordan; Royles, Jessica; Campbell, David I.; Keller, Elizabeth D.; Baisden, W. Troy; Roland, Thomas P.; Gallego-Sala, Angela V.

    2015-11-01

    Variations in the isotopic composition of precipitation are determined by fractionation processes which occur during temperature- and humidity-dependent phase changes associated with evaporation and condensation. Oxygen stable isotope ratios have therefore been frequently used as a source of palaeoclimate data from a variety of proxy archives, which integrate this signal over time. Applications from ombrotrophic peatlands, where the source water used in cellulose synthesis is derived solely from precipitation, have been mostly limited to Northern Hemisphere Sphagnum-dominated bogs, with few in the Southern Hemisphere or in peatlands dominated by vascular plants. New Zealand (NZ) provides an ideal location to undertake empirical research into oxygen isotope fractionation in vascular peatlands because single taxon analysis can be easily carried out, in particular using the preserved root matrix of the restionaceous wire rush (Empodisma spp.) that forms deep Holocene peat deposits throughout the country. Furthermore, large gradients are observed in the mean isotopic composition of precipitation across NZ, caused primarily by the relative influence of different climate modes. Here, we test whether δ18O of Empodisma α-cellulose from ombrotrophic restiad peatlands in NZ can provide a methodology for developing palaeoclimate records of past precipitation δ18O. Surface plant, water and precipitation samples were taken over spatial (six sites spanning >10° latitude) and temporal (monthly measurements over one year) gradients. A link between the isotopic composition of root-associated water, the most likely source water for plant growth, and precipitation in both datasets was found. Back-trajectory modelling of precipitation moisture source for rain days prior to sampling showed clear seasonality in the temporal data that was reflected in root-associated water. The link between source water and plant cellulose was less clear, although mechanistic modelling predicted mean

  10. Global Radiological Source Sorting, Tracking, and Monitoring (Gradsstram) Using Emergin RFID and Web 2.0 Technologies to Provide Total Asset and Information Visualization, Paper at 2009 INMM

    SciTech Connect

    Walker, Randy M.; Kopsick, Deborah A.; Gorman, Bryan L.; Ganguly, Auroop R.; Ferren, Mitch; Shankar, Mallikarjun

    2009-01-01

    Background Thousands of shipments of radioisotopes developed in the United States (U.S.) are transported domestically and internationally for medical and industrial applications, including to partner laboratories in European Union (EU) countries. Over the past five years, the Environmental Protection Agency (EPA), the Department of Energy (DOE), and Oak Ridge National Laboratory (ORNL)1 have worked with state first responder personnel, key private sector supply chain stakeholders, the Department of Homeland Security (DHS), the Department of Transportation (DOT), the Department of Defense (DoD) and the Nuclear Regulatory Commission (NRC) on Radio Frequency Identification (RFID) tracking and monitoring of medical, research and industrial radioisotopes in commerce. ORNL was the pioneer of the international radioisotope shipping and production business. Most radioisotopes made and used today were either made or discovered at ORNL. While most of the radioisotopes used in the commercial sector are now produced and sold by the private market, ORNL still leads the world in the production of exotic, high-value and/or sensitive industrial, medical and research isotopes. The ORNL-EPA-DOE Radiological Source Tracking and Monitoring (RadSTraM) project tested, evaluated, and integrated RFID technologies in laboratory settings and at multiple private-sector shipping and distribution facilities (Perkin Elmer and DHL) to track and monitor common radioisotopes used in everyday commerce. The RFID tracking capability was also tested in association with other deployed technologies including radiation detection, chemical/explosives detection, advanced imaging, lasers, and infrared scanning. At the 2007 EU-U.S. Summit, the leaders of the US Department of Commerce (DOC) and EU European Commission (EC) committed to pursue jointly directed Lighthouse Priority Projects. These projects are intended to foster cooperation and reduce regulatory burdens with respect to transatlantic commerce. The

  11. Pilot-Scale Pulsed UV Light Irradiation of Experimentally Infected Raspberries Suppresses Cryptosporidium parvum Infectivity in Immunocompetent Suckling Mice.

    PubMed

    Le Goff, L; Hubert, B; Favennec, L; Villena, I; Ballet, J J; Agoulon, A; Orange, N; Gargala, G

    2015-12-01

    Cryptosporidium spp., a significant cause of foodborne infection, have been shown to be resistant to most chemical food disinfectant agents and infective for weeks in irrigation waters and stored fresh vegetal produce. Pulsed UV light (PL) has the potential to inactivate Cryptosporidium spp. on surfaces of raw or minimally processed foods or both. The present study aimed to evaluate the efficacy of PL on viability and in vivo infectivity of Cryptosporidium parvum oocysts present on raspberries, a known source of transmission to humans of oocyst-forming apicomplexan pathogens. The skin of each of 20 raspberries was experimentally inoculated with five 10-μl spots of an oocyst suspension containing 6 × 10(7) oocysts per ml (Nouzilly isolate). Raspberries were irradiated by PL flashes (4 J/cm(2) of total fluence). This dose did not affect colorimetric or organoleptic characteristics of fruits. After immunomagnetic separation from raspberries, oocysts were bleached and administered orally to neonatal suckling mice. Seven days after infection, mice were euthanized, and the number of oocysts in the entire small intestine was individually assessed by immunofluorescence flow cytometry. Three of 12 and 12 of 12 inoculated mice that received 10 and 100 oocysts isolated from nonirradiated raspberries, respectively, were found infected. Four of 12 and 2 of 12 inoculated mice that received 10(3) and 10(4) oocysts from irradiated raspberries, respectively, were found infected. Oocyst counts were lower in animals inoculated with 10(3) and 10(4) oocysts from irradiated raspberries (92 ± 144 and 38 ± 82, respectively) than in animals infected with 100 oocysts from nonirradiated raspberries (35,785 ± 66,221, P = 0.008). PL irradiation achieved oocyst reductions of 2 and 3 log for an inoculum of 10(3) and 10(4) oocysts, respectively. The present pilot-scale evaluation suggests that PL is an effective mode of decontamination for raspberries and prompts further applicability

  12. Tracking the evolution of stream DOM source during storm events using end member mixing analysis based on DOM quality

    NASA Astrophysics Data System (ADS)

    Yang, Liyang; Chang, Soon-Woong; Shin, Hyun-Sang; Hur, Jin

    2015-04-01

    The source of river dissolved organic matter (DOM) during storm events has not been well constrained, which is critical in determining the quality and reactivity of DOM. This study assessed temporal changes in the contributions of four end members (weeds, leaf litter, soil, and groundwater), which exist in a small forested watershed (the Ehwa Brook, South Korea), to the stream DOM during two storm events, using end member mixing analysis (EMMA) based on spectroscopic properties of DOM. The instantaneous export fluxes of dissolved organic carbon (DOC), chromophoric DOM (CDOM), and fluorescent components were all enhanced during peak flows. The DOC concentration increased with the flow rate, while CDOM and humic-like fluorescent components were diluted around the peak flows. Leaf litter was dominant for the DOM source in event 2 with a higher rainfall, although there were temporal variations in the contributions of the four end members to the stream DOM for both events. The contribution of leaf litter peaked while that of deeper soils decreased to minima at peak flows. Our results demonstrated that EMMA based on DOM properties could be used to trace the DOM source, which is of fundamental importance for understanding the factors responsible for river DOM dynamics during storm events.

  13. TRACKED VEHICLE Rev 75

    SciTech Connect

    Raby, Eric Y.

    2007-05-08

    Revision 75 of the Tracked Vehicle software is a soft real-time simulation of a differentially steered, tracked mobile robot, which, because of the track flippers, resembles the iRobot PackBot (http://www.irobot.com/). Open source libraries are used for the physics engine (http://www.ode.org/), the display and user interface (http://www.mathies.com/cpw/), and the program command line and configuration file parameters (http://www.boost.org/). The simulation can be controlled by a USB joystick or the keyboard. The configuration file contains demonstration model parameters of no particular vehicle. This simulation can be used as a starting point for those doing tracked vehicle simulations. This simulation software is essentially a research tool which can be modified and adapted for certain types of tracked vehicle research. An open source license allows an individual researchers to tailor the code to their specific research needs.

  14. Characterizing the concentration of Cryptosporidium in Australian surface waters for setting health-based targets for drinking water treatment.

    PubMed

    Petterson, S; Roser, D; Deere, D

    2015-09-01

    It is proposed that the next revision of the Australian Drinking Water Guidelines will include 'health-based targets', where the required level of potable water treatment quantitatively relates to the magnitude of source water pathogen concentrations. To quantify likely Cryptosporidium concentrations in southern Australian surface source waters, the databases for 25 metropolitan water supplies with good historical records, representing a range of catchment sizes, land use and climatic regions were mined. The distributions and uncertainty intervals for Cryptosporidium concentrations were characterized for each site. Then, treatment targets were quantified applying the framework recommended in the World Health Organization Guidelines for Drinking-Water Quality 2011. Based on total oocyst concentrations, and not factoring in genotype or physiological state information as it relates to infectivity for humans, the best estimates of the required level of treatment, expressed as log10 reduction values, ranged among the study sites from 1.4 to 6.1 log10. Challenges associated with relying on historical monitoring data for defining drinking water treatment requirements were identified. In addition, the importance of quantitative microbial risk assessment input assumptions on the quantified treatment targets was investigated, highlighting the need for selection of locally appropriate values.

  15. Genetic Diversity of Cryptosporidium spp. within a Remote Population of Soay Sheep on St. Kilda Islands, Scotland

    PubMed Central

    Connelly, L.; Craig, B. H.; Jones, B.

    2013-01-01

    This is the first report to characterize the genotypes and subtypes of Cryptosporidium species infecting a geographically isolated population of feral Soay sheep (Ovis aries) on Hirta, St. Kilda, Scotland, during two distinct periods: (i) prior to a population crash and (ii) as host numbers increased. Cryptosporidium DNA was extracted by freeze-thawing of immunomagnetically separated (IMS) bead-oocyst complexes, and species were identified following nested-PCR-restriction fragment length polymorphism (RFLP)/PCR sequencing at two Cryptosporidium 18S rRNA loci. Two hundred fifty-five samples were analyzed, and the prevalent Cryptosporidium species in single infections were identified as C. hominis (11.4% of all samples tested), C. parvum (9%), C. xiaoi (12.5%), and C. ubiquitum (6.7%). Cryptosporidium parvum was also present with other Cryptosporidium species in 27.1% of all samples tested. Cryptosporidium parvum- and C. hominis-positive isolates were genotyped using two nested-PCR assays that amplify the Cryptosporidium glycoprotein 60 gene (GP60). GP60 gene analysis showed the presence of two Cryptosporidium genotypes, namely, C. parvum IIaA19G1R1 and C. hominis IbA10G2. This study reveals a higher diversity of Cryptosporidium species/genotypes than was previously expected. We suggest reasons for the high diversity of Cryptosporidium parasites within this isolated population and discuss the implications for our understanding of cryptosporidiosis. PMID:23354707

  16. Development and validation of a microbial source tracking marker for the detection of fecal pollution by muskrats.

    PubMed

    Marti, Romain; Zhang, Yun; Lapen, David R; Topp, Edward

    2011-10-01

    Surface waters in mixed-activity watersheds can be compromised by fecal pollution from livestock production, sewage outflows or leaking septic systems, or avian or mammalian wildlife. While undertaking water quality research in Eastern Ontario Canada, we observed an abundance of muskrats (Ondatra zibethicus) in some waterways, and sought to determine their significance with respect to fecal pollution loads. To that end, we developed and validated here a sensitive and specific marker, designated MuBa01, based on the 16S rRNA gene that distinguishes muskrat feces from human, livestock, or other wildlife sources. DNA from waterways inhabited by muskrats carried the marker. The widely used pig-specific marker Pig-2-Bac yielded a product with some muskrat feces, suggesting that it lacks specificity required to establish the significance of swine production in areas inhabited by muskrats. Overall, the MuBa01 marker will be useful for distinguishing the significance of this wide-ranging semi-aquatic mammal in the context of other fecal pollution sources.

  17. Well water quality in rural Nicaragua using a low-cost bacterial test and microbial source tracking.

    PubMed

    Weiss, Patricia; Aw, Tiong Gim; Urquhart, Gerald R; Galeano, Miguel Ruiz; Rose, Joan B

    2016-04-01

    Water-related diseases, particularly diarrhea, are major contributors to morbidity and mortality in developing countries. Monitoring water quality on a global scale is crucial to making progress in terms of population health. Traditional analytical methods are difficult to use in many regions of the world in low-resource settings that face severe water quality issues due to the inaccessibility of laboratories. This study aimed to evaluate a new low-cost method (the compartment bag test (CBT)) in rural Nicaragua. The CBT was used to quantify the presence of Escherichia coli in drinking water wells and aimed to determine the source(s) of any microbial contamination. Results indicate that the CBT is a viable method for use in remote rural regions. The overall quality of well water in Pueblo Nuevo, Nicaragua was deemed unsafe, and results led to the conclusion that animal fecal wastes may be one of the leading causes of well contamination. Elevation and depth of wells were not found to impact overall water quality. However rope-pump wells had a 64.1% reduction in contamination when compared with simple wells.

  18. Molecular Characterization of Cryptosporidium Species and Giardia duodenalis from Symptomatic Cambodian Children

    PubMed Central

    Moore, Catrin E.; Elwin, Kristin; Phot, Nget; Seng, Chanthou; Mao, Saroeun; Suy, Kuong; Kumar, Varun; Nader, Johanna; Bousfield, Rachel; Perera, Sanuki; Bailey, J. Wendi; Beeching, Nicholas J.; Day, Nicholas P. J.; Parry, Christopher M.; Chalmers, Rachel M.

    2016-01-01

    Background In a prospective study, 498 single faecal samples from children aged under 16 years attending an outpatient clinic in the Angkor Hospital for Children, northwest Cambodia, were examined for Cryptosporidium oocysts and Giardia cysts using microscopy and molecular assays. Methodology/Principal Findings Cryptosporidium oocysts were detected in 2.2% (11/498) of samples using microscopy and in 7.7% (38/498) with molecular tests. Giardia duodenalis cysts were detected in 18.9% (94/498) by microscopy and 27.7% (138/498) by molecular tests; 82% of the positive samples (by either method) were from children aged 1–10 years. Cryptosporidium hominis was the most common species of Cryptosporidium, detected in 13 (34.2%) samples, followed by Cryptosporidium meleagridis in 9 (23.7%), Cryptosporidium parvum in 8 (21.1%), Cryptosporidium canis in 5 (13.2%), and Cryptosporidium suis and Cryptosporidium ubiquitum in one sample each. Cryptosporidium hominis and C. parvum positive samples were subtyped by sequencing the GP60 gene: C. hominis IaA16R6 and C. parvum IIeA7G1 were the most abundant subtypes. Giardia duodenalis was typed using a multiplex real-time PCR targeting assemblages A and B. Assemblage B (106; 76.8% of all Giardia positive samples) was most common followed by A (12.3%) and mixed infections (5.1%). Risk factors associated with Cryptosporidium were malnutrition (AOR 9.63, 95% CI 1.67–55.46), chronic medical diagnoses (AOR 4.51, 95% CI 1.79–11.34) and the presence of birds in the household (AOR 2.99, 95% CI 1.16–7.73); specifically C. hominis (p = 0.03) and C. meleagridis (p<0.001) were associated with the presence of birds. The use of soap was protective against Giardia infection (OR 0.74, 95% CI 0.58–0.95). Conclusions/Significance This is the first report to describe the different Cryptosporidium species and subtypes and Giardia duodenalis assemblages in Cambodian children. The variety of Cryptosporidium species detected indicates both

  19. Molecular-based investigation of Cryptosporidium and Giardia from animals in water catchments in southeastern Australia.

    PubMed

    Nolan, Matthew J; Jex, Aaron R; Koehler, Anson V; Haydon, Shane R; Stevens, Melita A; Gasser, Robin B

    2013-04-01

    There has been no large-scale systematic molecular epidemiological investigation of the waterborne protozoans, Cryptosporidium or Giardia, in southeastern Australia. Here, we explored, for the first time, the genetic composition of these genera in faecal samples from animals in nine Melbourne Water reservoir areas, collected over a period of two-years. We employed PCR-based single-strand conformation polymorphism (SSCP) and phylogenetic analyses of loci (pSSU and pgp60) in the small subunit (SSU) of ribosomal RNA and 60-kDa glycoprotein (gp60) genes to detect and characterise Cryptosporidium, and another locus (ptpi) in the triose-phosphate isomerase (tpi) gene to identify and characterise Giardia. Cryptosporidium was detected in 2.8% of the 2009 samples examined; the analysis of all amplicons defined 14 distinct sequence types for each of pSSU and pgp60, representing Cryptosporidium hominis (genotype Ib - subgenotype IbA10G2R2), Cryptosporidium parvum (genotype IIa - subgenotypes IIaA15G2R1, IIaA19G2R1, IIaA19G3R1, IIaA19G4R1, IIaA20G3R1, IIaA20G4R1, IIaA20G3R2 and IIaA21G3R1), Cryptosporidium cuniculus (genotype Vb - subgenotypes VbA22R4, VbA23R3, VbA24R3, VbA25R4 and VbA26R4), and Cryptosporidium canis, Cryptosporidium fayeri, Cryptosporidium macropodum and Cryptosporidium ubiquitum as well as six new pSSU sequence types. In addition, Giardia was identified in 3.4% of the samples; all 28 distinct ptpi sequence types defined were linked to assemblage A of Giardia duodenalis. Of all 56 sequence types characterised, eight and one have been recorded previously in Cryptosporidium and Giardia, respectively, from humans. In contrast, nothing is known about the zoonotic potential of 35 new genotypes of Cryptosporidium and Giardia recorded here for the first time. Future work aims to focus on estimating the prevalence of Cryptosporidium and Giardia genotypes in humans and a wide range of animals in Victoria and elsewhere in Australia. (Nucleotide sequences reported in

  20. Characterization of Enterococcus spp. from human and animal feces using 16S rRNA sequences, the esp gene, and PFGE for microbial source tracking in Korea.

    PubMed

    Kim, Sei-Yoon; Lee, Jung Eun; Lee, Sunghee; Lee, Hee Tae; Hur, Ho-Gil; Ko, Gwangpyo

    2010-05-01

    Contamination from human and animal fecal waste is a primary cause of water pollution. Microbial source tracking (MST) may be a useful tool for high-quality environmental management and for assessing human health risks associated with water pollution. The goal of this study was to evaluate Enterococcus spp. as a target organism for MST. Thirty-four fecal samples were collected from five different sources (human, chicken, pig, cow, and goose) in South Korea. In total, 237 Enterococcus spp. were isolated from feces using membrane- Enterococcus indoxyl-beta-d-glucoside agar. The 16S rRNA gene and the whole genome were analyzed using nucleic acid sequencing and pulsed-field gel electrophoresis (PFGE), respectively. Both phylogenetic analysis and principal coordinate analysis using UniFrac were performed on the nucleic acid sequences of the 16S rRNA gene. According to P-tests from UniFrac, significant differences existed between Enterococcus spp. isolated from human feces and those from animal feces. In addition, we evaluated whether the esp gene of Enterococcus faecium could be a specific target for Enterococcus spp. isolated from human feces. Of 58 E. faecium isolates tested, only three were esp-positive. The specificity of the esp gene of E. faecium isolated from human feces was 100%, but the sensitivity was <10%. These results suggest that Enterococcus spp. have different molecular characteristics according to their fecal source and that these characteristics can be further identified by analyzing the esp gene and 16S rRNA sequences, whereas PFGE provides limited information on the fecal sources of Enterococcus spp.

  1. Application of F⁺RNA Coliphages as Source Tracking Enteric Viruses on Parsley and Leek Using RT-PCR.

    PubMed

    Shahrampour, Dina; Yavarmanesh, Masoud; Najafi, Mohammad Bagher Habibi; Mohebbi, Mohebbat

    2015-12-01

    The objective of this study was to identify sources of fecal contamination in leek and parsley, by using four different F(+)RNA coliphage genogroups (IV, I indicate animal fecal contamination and II, III indicate human fecal contamination). Three different concentrations (10(2), 10(4), 10(6) pfu/ml) of MS2 coliphage were inoculated on the surface of parsley and leek samples for detection of phage recovery efficiency among two methods of elution concentration (PEG-precipitation and Ultracentrifugation) by performing double agar layer (DAL) assay in three replications. Highest recovery of MS2 was observed in PEG method and in 10(6) inoculation concentration. Accordingly, the PEG method was used for washing and isolation of potentially contaminated phages of 30 collected samples (15 samples from the market and 15 samples from the farm). The final solutions of PEG method were tested for the enumeration of plaques by DAL assay. Total RNA was then extracted from recovered phages, and RT-PCR was performed by using four primer sets I, II, III, and IV. Incidence of F(+)RNA coliphages was observed in 12/15 (80 %) and 10/15 (66/6 %) of samples were obtained from farm and market, respectively, using both DAL and RT-PCR test methods. Different genotypes (I, II, and IV) of F(+)RNA coliphages were found in farm samples, while only genotype I was detected in market samples by using the primer sets. Due to the higher frequency of genotype I and IV, the absence of genotype III, and also the low frequency of genotype II, it is concluded that the contamination of vegetable (parsley and leek) in Neyshabour, Iran is most likely originated from animal sources.

  2. Assessment of Genetic Markers for Tracking the Sources of Human Wastewater Associated Escherichia coli in Environmental Waters.

    PubMed

    Warish, Ahmed; Triplett, Cheryl; Gomi, Ryota; Gyawali, Pradip; Hodgers, Leonie; Toze, Simon

    2015-08-04

    In this study, we have evaluated the performance characteristics (host-specificity and -sensitivity) of four human wastewater-associated Escherichia coli (E. coli) genetic markers (H8, H12, H14, and H24) in 10 target (human) and nontarget (cat, cattle, deer, dog, emu, goat, horse, kangaroo, and possum) host groups in Southeast Queensland, Australia. The overall host-sensitivity values of the tested markers in human wastewater samples were 1.0 (all human wastewater samples contained the E. coli genetic marke