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Sample records for cryptosporidium source tracking

  1. Tracking host sources of Cryptosporidium spp. in raw water for improved health risk assessment.

    PubMed

    Ruecker, Norma J; Braithwaite, Shannon L; Topp, Edward; Edge, Thomas; Lapen, David R; Wilkes, Graham; Robertson, Will; Medeiros, Diane; Sensen, Christoph W; Neumann, Norman F

    2007-06-01

    Recent molecular evidence suggests that different species and/or genotypes of Cryptosporidium display strong host specificity, altering our perceptions regarding the zoonotic potential of this parasite. Molecular forensic profiling of the small-subunit rRNA gene from oocysts enumerated on microscope slides by U.S. Environmental Protection Agency method 1623 was used to identify the range and prevalence of Cryptosporidium species and genotypes in the South Nation watershed in Ontario, Canada. Fourteen sites within the watershed were monitored weekly for 10 weeks to assess the occurrence, molecular composition, and host sources of Cryptosporidium parasites impacting water within the region. Cryptosporidium andersoni, Cryptosporidium muskrat genotype II, Cryptosporidium cervine genotype, C. baileyi, C. parvum, Cryptosporidium muskrat genotype I, the Cryptosporidium fox genotype, genotype W1, and genotype W12 were detected in the watershed. The molecular composition of the Cryptosporidium parasites, supported by general land use analysis, indicated that mature cattle were likely the main source of contamination of the watershed. Deer, muskrats, voles, birds, and other wildlife species, in addition to sewage (human or agricultural) may also potentially impact water quality within the study area. Source water protection studies that use land use analysis with molecular genotyping of Cryptosporidium parasites may provide a more robust source-tracking tool to characterize fecal impacts in a watershed. Moreover, the information is vital for assessing environmental and human health risks posed by water contaminated with zoonotic and/or anthroponotic forms of Cryptosporidium. PMID:17483276

  2. Cryptosporidium Source Tracking in the Potomac River Watershed▿

    PubMed Central

    Yang, Wenli; Chen, Plato; Villegas, Eric N.; Landy, Ronald B.; Kanetsky, Charles; Cama, Vitaliano; Dearen, Theresa; Schultz, Cherie L.; Orndorff, Kenneth G.; Prelewicz, Gregory J.; Brown, Miranda H.; Young, Kim Roy; Xiao, Lihua

    2008-01-01

    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 in the watershed. These sites included two water treatment plant intakes, as well as three upstream sites, each associated with a different type of land use. The uses, including urban wastewater, agricultural (cattle) wastewater, and wildlife, posed different risks in terms of the potential contribution of Cryptosporidium oocysts to the source water. Cryptosporidium was detected in 27 base flow water samples and 23 storm flow water samples. The most frequently detected species was C. andersoni (detected in 41 samples), while 14 other species or genotypes, almost all wildlife associated, were occasionally detected. The two common human-pathogenic species, C. hominis and C. parvum, were not detected. Although C. andersoni was common at all four sites influenced by agriculture, it was largely absent at the urban wastewater site. There were very few positive samples as determined by Environmental Protection Agency method 1623 at any site; only 8 of 90 samples analyzed (9%) were positive for Cryptosporidium as determined by microscopy. The genotyping results suggest that many of the Cryptosporidium oocysts in the water treatment plant source waters were from old calves and adult cattle and might not pose a significant risk to human health. PMID:18776033

  3. 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...

  4. 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...

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

    PubMed Central

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

    2015-01-01

    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. PMID:26092455

  6. Tracking Cryptosporidium parvum by sequence analysis of small double-stranded RNA.

    PubMed Central

    Xiao, L.; Limor, J.; Bern, C.; Lal, A. A.

    2001-01-01

    We sequenced a 173-nucleotide fragment of the small double-stranded viruslike RNA of Cryptosporidium parvum isolates from 23 calves and 38 humans. Sequence diversity was detected at 17 sites. Isolates from the same outbreak had identical double-stranded RNA sequences, suggesting that this technique may be useful for tracking Cryptosporidium infection sources. PMID:11266306

  7. 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.

  8. 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. PMID:19603633

  9. Molecular and phylogenetic approaches for assessing sources of Cryptosporidium contamination in water.

    PubMed

    Ruecker, Norma J; Matsune, Joanne C; Wilkes, Graham; Lapen, David R; Topp, Edward; Edge, Thomas A; Sensen, Christoph W; Xiao, Lihua; Neumann, Norman F

    2012-10-15

    The high sequence diversity and heterogeneity observed within species or genotypes of Cryptosporidium requires phylogenetic approaches for the identification of novel sequences obtained from the environment. A long-term study on Cryptosporidium in the agriculturally-intensive South Nation River watershed in Ontario, Canada was undertaken, in which 60 sequence types were detected. Of these sequence types 33 were considered novel with no identical matches in GenBank. Detailed phylogenetic analysis identified that most sequences belonged to 17 previously described species: Cryptosporidium andersoni, Cryptosporidium baileyi, Cryptosporidium hominis, Cryptosporidium parvum, Cryptosporidium ubiquitum, Cryptosporidium meleagridis, muskrat I, muskrat II, deer mouse II, fox, vole, skunk, shrew, W12, W18, W19 and W25 genotypes. In addition, two new genotypes were identified, W27 and W28. C. andersoni and the muskrat II genotype were most frequently detected in the water samples. Species associated with livestock made up 39% of the total molecular detections, while wildlife associated species and genotypes accounted for 55% of the Cryptosporidium identified. The human pathogenic species C. hominis and C. parvum had an overall prevalence of 1.6% in the environment, indicating a small risk to humans from the Cryptosporidium present in the watershed. Phylogenetic analysis and knowledge of host-parasite relationships are fundamental in using Cryptosporidium as a source-tracking or human health risk assessment tool. PMID:22841595

  10. Monitoring of Cryptosporidium and Giardia in Czech drinking water sources.

    PubMed

    Dolejs, P; Ditrich, O; Machula, T; Kalousková, N; Puzová, G

    2000-01-01

    In Czech raw water sources for drinking water supply, Cryptosporidium was found in numbers from 0 to 7400 per 100 liters and Giardia from 0 to 485 per 100 liters. The summer floods of 1997 probably brought the highest numbers of Cryptosporidium oocysts into one of the reservoirs sampled; since then these numbers decreased steadily. A relatively high number of Cryptosporidium oocysts was found in one sample of treated water. Repeated sampling demonstrated that this was a sporadic event. The reason for the presence of Cryptosporidium in a sample of treated drinking-water is unclear and requires further study.

  11. 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...

  12. Cryptosporidium

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Of nearly 25 named species and numerous genotypes of Cryptosporidium, two are of special importance relative to human health and food safety: Cryptosporidium hominis and Cryptosporidium parvum, the former with a predilection for humans and the latter a promiscuous species. Genetic tools have been es...

  13. 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...

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  19. 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...

  20. Occurrence, Source, and Human Infection Potential of Cryptosporidium and Giardia spp. in Source and Tap Water in Shanghai, China▿

    PubMed Central

    Feng, Yaoyu; Zhao, Xukun; Chen, Jiaxu; Jin, Wei; Zhou, Xiaonong; Li, Na; Wang, Lin; Xiao, Lihua

    2011-01-01

    Genotyping studies on the source and human infection potential of Cryptosporidium oocysts in water have been almost exclusively conducted in industrialized nations. In this study, 50 source water samples and 30 tap water samples were collected in Shanghai, China, and analyzed by the U.S. Environmental Protection Agency (EPA) Method 1623. To find a cost-effective method to replace the filtration procedure, the water samples were also concentrated by calcium carbonate flocculation (CCF). Of the 50 source water samples, 32% were positive for Cryptosporidium and 18% for Giardia by Method 1623, whereas 22% were positive for Cryptosporidium and 10% for Giardia by microscopy of CCF concentrates. When CCF was combined with PCR for detection, the occurrence of Cryptosporidium (28%) was similar to that obtained by Method 1623. Genotyping of Cryptosporidium in 17 water samples identified the presence of C. andersoni in 14 water samples, C. suis in 7 water samples, C. baileyi in 2 water samples, C. meleagridis in 1 water sample, and C. hominis in 1 water sample. Therefore, farm animals, especially cattle and pigs, were the major sources of water contamination in Shanghai source water, and most oocysts found in source water in the area were not infectious to humans. Cryptosporidium oocysts were found in 2 of 30 tap water samples. The combined use of CCF for concentration and PCR for detection and genotyping provides a less expensive alternative to filtration and fluorescence microscopy for accurate assessment of Cryptosporidium contamination in water, although the results from this method are semiquantitative. PMID:21498768

  1. 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

  2. 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

  3. Environmental sources of Cryptosporidium in an urban slum in northeastern Brazil.

    PubMed

    Newman, R D; Wuhib, T; Lima, A A; Guerrant, R L; Sears, C L

    1993-08-01

    Cryptosporidium is an important cause of diarrheal disease in children worldwide. To elucidate the environmental sources of this parasite, we selected an urban slum in Fortaleza, Brazil, a community with a known high incidence of cryptosporidiosis, and examined both stool smears from household animals (n = 127) and filtrates from local water sources (n = 18) for Cryptosporidium oocysts. Because previous work in this community has demonstrated the seasonal nature of human infection with Cryptosporidium, collections were made separately for the dry and rainy seasons. Of the 64 stools collected during the dry season (September-December 1990), four (6.3%) were positive by acid-fast staining for Cryptosporidium. Of the 63 rainy season samples (March-May 1991), nine (14.3%) were positive. Overall, oocysts were detected in 13 (10.2%) of 127 animal stool samples. Freshwater samples were obtained from a variety of sources including open and closed wells, and running city water and then processed. Four of 18 samples (22.2%), including a sample from city water were positive by at least one of two staining techniques (acid-fast and immunofluorescence). In summary, animals may serve as a reservoir of Cryptosporidium, with potential for the contamination of immediate household water sources. These findings may help to explain the high incidence of cryptosporidiosis among infants in this impoverished community.

  4. 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...

  5. 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...

  6. 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...

  7. 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. PMID:25306487

  8. 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.

  9. Cryptosporidium Taxonomy: Recent Advances and Implications for Public Health

    PubMed Central

    Xiao, Lihua; Fayer, Ronald; Ryan, Una; Upton, Steve J.

    2004-01-01

    There has been an explosion of descriptions of new species of Cryptosporidium during the last two decades. This has been accompanied by confusion regarding the criteria for species designation, largely because of the lack of distinct morphologic differences and strict host specificity among Cryptosporidium spp. A review of the biologic species concept, the International Code of Zoological Nomenclature (ICZN), and current practices for Cryptosporidium species designation calls for the establishment of guidelines for naming Cryptosporidium species. All reports of new Cryptosporidium species should include at least four basic components: oocyst morphology, natural host specificity, genetic characterizations, and compliance with the ICZN. Altogether, 13 Cryptosporidium spp. are currently recognized: C. muris, C. andersoni, C. parvum, C. hominis, C. wrairi, C. felis, and C. cannis in mammals; C. baïleyi, C. meleagridis, and C. galli in birds; C. serpentis and C. saurophilum in reptiles; and C. molnari in fish. With the establishment of a framework for naming Cryptosporidium species and the availability of new taxonomic tools, there should be less confusion associated with the taxonomy of the genus Cryptosporidium. The clarification of Cryptosporidium taxonomy is also useful for understanding the biology of Cryptosporidium spp., assessing the public health significance of Cryptosporidium spp. in animals and the environment, characterizing transmission dynamics, and tracking infection and contamination sources. PMID:14726456

  10. 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...

  11. 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 at the (treatment plant name) is sufficient to adequately remove Cryptosporidium from your...

  12. 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 at the (treatment plant name) is sufficient to adequately remove Cryptosporidium from your...

  13. 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...

  14. Comparison of Method 1623 and Cell Culture-PCR for Detection of Cryptosporidium spp. in Source Waters

    PubMed Central

    LeChevallier, Mark W.; Di Giovanni, George D.; Clancy, Jennifer L.; Bukhari, Zia; Bukhari, Shan; Rosen, Jeffrey S.; Sobrinho, Jose; Frey, Michelle M.

    2003-01-01

    Analysis of Cryptosporidium occurrence in six watersheds by method 1623 and the integrated cell culture-PCR (CC-PCR) technique provided an opportunity to evaluate these two methods. The average recovery efficiencies were 58.5% for the CC-PCR technique and 72% for method 1623, but the values were not significantly different (P = 0.06). Cryptosporidium oocysts were detected in 60 of 593 samples (10.1%) by method 1623. Infectious oocysts were detected in 22 of 560 samples (3.9%) by the CC-PCR technique. There was 87% agreement between the total numbers of samples positive as determined by method 1623 and CC-PCR for four of the sites. The other two sites had 16.3 and 24% correspondence between the methods. Infectious oocysts were detected in all of the watersheds. Overall, approximately 37% of the Cryptosporidium oocysts detected by the immunofluorescence method were viable and infectious. DNA sequence analysis of the Cryptosporidium parvum isolates detected by CC-PCR showed the presence of both the bovine and human genotypes. More than 90% of the C. parvum isolates were identified as having the bovine or bovine-like genotype. The estimates of the concentrations of infectious Cryptosporidium and the resulting daily and annual risks of infection compared well for the two methods. The results suggest that most surface water systems would require, on average, a 3-log reduction in source water Cryptosporidium levels to meet potable water goals. PMID:12571019

  15. Comparison of method 1623 and cell culture-PCR for detection of Cryptosporidium spp. in source waters.

    PubMed

    LeChevallier, Mark W; Di Giovanni, George D; Clancy, Jennifer L; Bukhari, Zia; Bukhari, Shan; Rosen, Jeffrey S; Sobrinho, Jose; Frey, Michelle M

    2003-02-01

    Analysis of Cryptosporidium occurrence in six watersheds by method 1623 and the integrated cell culture-PCR (CC-PCR) technique provided an opportunity to evaluate these two methods. The average recovery efficiencies were 58.5% for the CC-PCR technique and 72% for method 1623, but the values were not significantly different (P = 0.06). Cryptosporidium oocysts were detected in 60 of 593 samples (10.1%) by method 1623. Infectious oocysts were detected in 22 of 560 samples (3.9%) by the CC-PCR technique. There was 87% agreement between the total numbers of samples positive as determined by method 1623 and CC-PCR for four of the sites. The other two sites had 16.3 and 24% correspondence between the methods. Infectious oocysts were detected in all of the watersheds. Overall, approximately 37% of the Cryptosporidium oocysts detected by the immunofluorescence method were viable and infectious. DNA sequence analysis of the Cryptosporidium parvum isolates detected by CC-PCR showed the presence of both the bovine and human genotypes. More than 90% of the C. parvum isolates were identified as having the bovine or bovine-like genotype. The estimates of the concentrations of infectious Cryptosporidium and the resulting daily and annual risks of infection compared well for the two methods. The results suggest that most surface water systems would require, on average, a 3-log reduction in source water Cryptosporidium levels to meet potable water goals.

  16. Cryptosporidium: Prevention - Immunocompromised Persons

    MedlinePlus

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

  17. 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.

  18. Tracking Steady Light Sources Amid Luminous Transients

    NASA Technical Reports Server (NTRS)

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

    1994-01-01

    The Transient Event Rejection for Acquisition and Tracking (TERAT) algorithm governs operation of image-data-acquisition and -processing system. TERAT processes digitized image data to acquire (that is, identify) candidate steady source of light, validate candidate source, and track validated source, all in presence of real or apparent luminous transients represented in image data. Source of light tracked could be star or distant luminous beacon. Transients caused by impacts of ionizing radiation on imaging array of photodetectors or by unsteady light sources not meant to be tracked. TERAT functions with limited data-processing resources. TERAT algorithm currently operational on NASA's TOPEX mission.

  19. Attentive Tracking of Sound Sources.

    PubMed

    Woods, Kevin J P; McDermott, Josh H

    2015-08-31

    Auditory scenes often contain concurrent sound sources, but listeners are typically interested in just one of these and must somehow select it for further processing. One challenge is that real-world sounds such as speech vary over time and as a consequence often cannot be separated or selected based on particular values of their features (e.g., high pitch). Here we show that human listeners can circumvent this challenge by tracking sounds with a movable focus of attention. We synthesized pairs of voices that changed in pitch and timbre over random, intertwined trajectories, lacking distinguishing features or linguistic information. Listeners were cued beforehand to attend to one of the voices. We measured their ability to extract this cued voice from the mixture by subsequently presenting the ending portion of one voice and asking whether it came from the cued voice. We found that listeners could perform this task but that performance was mediated by attention-listeners who performed best were also more sensitive to perturbations in the cued voice than in the uncued voice. Moreover, the task was impossible if the source trajectories did not maintain sufficient separation in feature space. The results suggest a locus of attention that can follow a sound's trajectory through a feature space, likely aiding selection and segregation amid similar distractors. PMID:26279234

  20. 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...

  1. Effect of particles on the recovery of cryptosporidium oocysts from source water samples of various turbidities.

    PubMed

    Feng, Yao Yu; Ong, Say Leong; Hu, Jiang Yong; Song, Lian Fa; Tan, Xiao Lan; Ng, Wun Jern

    2003-04-01

    Cryptosporidium parvum can be found in both source and drinking water and has been reported to cause serious waterborne outbreaks which threaten public health safety. The U.S. Environmental Protection Agency has developed method 1622 for detection of Cryptosporidium oocysts present in water. Method 1622 involves four key processing steps: filtration, immunomagnetic separation (IMS), fluorescent-antibody (FA) staining, and microscopic evaluation. The individual performance of each of these four steps was evaluated in this study. We found that the levels of recovery of C. parvum oocysts at the IMS-FA and FA staining stages were high, averaging more than 95%. In contrast, the level of recovery declined significantly, to 14.4%, when the filtration step was incorporated with tap water as a spiking medium. This observation suggested that a significant fraction of C. parvum oocysts was lost during the filtration step. When C. parvum oocysts were spiked into reclaimed water, tap water, microfiltration filtrate, and reservoir water, the highest mean level of recovery of (85.0% +/- 5.2% [mean +/- standard deviation]) was obtained for the relatively turbid reservoir water. Further studies indicated that it was the suspended particles present in the reservoir water that contributed to the enhanced C. parvum oocyst recovery. The levels of C. parvum oocyst recovery from spiked reservoir water with different turbidities indicated that particle size and concentration could affect oocyst recovery. Similar observations were also made when silica particles of different sizes and masses were added to seeded tap water. The optimal particle size was determined to be in the range from 5 to 40 micro m, and the corresponding optimal concentration of suspended particles was 1.42 g for 10 liters of tap water.

  2. 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.

  3. Identification of Species and Sources of Cryptosporidium Oocysts in Storm Waters with a Small-Subunit rRNA-Based Diagnostic and Genotyping Tool

    PubMed Central

    Xiao, Lihua; Alderisio, Kerri; Limor, Josef; Royer, Michael; Lal, Altaf A.

    2000-01-01

    The identification of Cryptosporidium oocysts in environmental samples is largely made by the use of an immunofluorescent assay. In this study, we have used a small-subunit rRNA-based PCR-restriction fragment length polymorphism technique to identify species and sources of Cryptosporidium oocysts present in 29 storm water samples collected from a stream in New York. A total of 12 genotypes were found in 27 positive samples; for 4 the species and probable origins were identified by sequence analysis, whereas the rest represent new genotypes from wildlife. Thus, this technique provides an alternative method for the detection and differentiation of Cryptosporidium parasites in environmental samples. PMID:11097935

  4. Integrated Cryptosporidium Assay To Determine Oocyst Density, Infectivity, and Genotype for Risk Assessment of Source and Reuse Water

    PubMed Central

    King, Brendon; Fanok, Stella; Phillips, Renae; Swaffer, Brooke

    2015-01-01

    Cryptosporidium continues to be problematic for the water industry, with risk assessments often indicating that treatment barriers may fail under extreme conditions. However, risk analyses have historically used oocyst densities and not considered either oocyst infectivity or species/genotype, which can result in an overestimation of risk if the oocysts are not human infective. We describe an integrated assay for determining oocyst density, infectivity, and genotype from a single-sample concentrate, an important advance that overcomes the need for processing multiple-grab samples or splitting sample concentrates for separate analyses. The assay incorporates an oocyst recovery control and is compatible with standard primary concentration techniques. Oocysts were purified from primary concentrates using immunomagnetic separation prior to processing by an infectivity assay. Plate-based cell culture was used to detect infectious foci, with a monolayer washing protocol developed to allow recovery and enumeration of oocysts. A simple DNA extraction protocol was developed to allow typing of any wells containing infectious Cryptosporidium. Water samples from a variety of source water and wastewater matrices, including a semirural catchment, wastewater, an aquifer recharge site, and storm water, were analyzed using the assay. Results demonstrate that the assay can reliably determine oocyst densities, infectivity, and genotype from single-grab samples for a variety of water matrices and emphasize the varying nature of Cryptosporidium risk extant throughout source waters and wastewaters. This assay should therefore enable a more comprehensive understanding of Cryptosporidium risk for different water sources, assisting in the selection of appropriate risk mitigation measures. PMID:25769833

  5. 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. PMID:27310009

  6. 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.

  7. 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

  8. 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...

  9. 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...

  10. 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.

  11. Total and infectious Cryptosporidium oocyst and total Giardia cyst concentrations from distinct agricultural and urban contamination sources in Eastern Canada.

    PubMed

    Lalancette, Cindy; Généreux, Mylène; Mailly, Jacinthe; Servais, Pierre; Côté, Caroline; Michaud, Aubert; Di Giovanni, George D; Prévost, Michèle

    2012-03-01

    Cryptosporidium and Giardia (oo)cyst concentrations are frequently used for assessing drinking water safety. The widely used USEPA Method 1623 provides total counts of (oo)cysts, but may not be accurate for human health risk characterization, since it does not provide infectivity information. The total counts and infectious fraction of Cryptosporidium oocysts and the total counts of Giardia cysts were assessed in major fecal pollution sources. Fresh calf and cow feces, their manure, and the discharge point were sampled in a small rural sub-watershed (n = 20, 21, 10, 10). Median concentrations for total (oo)cysts were higher in calves (333 oocysts g(-1); 111 cysts g(-1)) than in cows (52 oocysts g(-1); 7 cysts g(-1)). Infectious oocysts were found in 17 (7%) of the samples and none were found in manure or at the discharge point. Urban sources were sampled in the influent and effluent (n = 19, 18) of two wastewater treatment plants. Peak concentrations were 533 oocysts L(-1) and 9,010 cysts L(-1) for influents and 89 oocysts L(-1) and 472 cysts L(-1) for effluents. Infectious oocyst fractions varied from below the detection limit to 7-22% in the effluent and influent respectively. These infectious fractions are significantly lower than those currently used for quantitative microbial risk assessment estimates. PMID:22361710

  12. Frequency of Cryptosporidium spp. as cause of human gastrointestinal disease in Switzerland and possible sources of infection.

    PubMed

    Baumgartner, A; Marder, H P; Munzinger, J; Siegrist, H H

    2000-09-01

    Detection of Cryptosporidium parvum is not routinely done in laboratories of clinical microbiology and there is no obligation to communicate isolations of this pathogen to health authorities. For these reasons, frequency of cryptosporidiosis and sources of infection are only poorly known in Switzerland. To obtain more concise information in this field, feces from 5179 hospitalized and 1256 ambulatory patients with suspected gastrointestinal infections were screened for Cryptosporidium spp. over the period of one year in two laboratories. In toto, 13 patients with cryptosporidiosis were detected which results a frequency of 0.2%. Furthermore, it was shown by a projection that about 340 cases of cryptosporidiosis have to expected yearly in Switzerland, resulting an estimated morbidity of 4.85 cases per 100,000 persons. With regard to risk factors, the available patient data did not allow solid statistical conclusions. However, known risk factors such as immunosuppression, travelling abroad (33.3%) and contact to symptomatic persons were unquestionably demonstrated. Oysters, raw milk and cream from raw milk had to be strongly taken into consideration as food vehicles of transmission. Tap water from municipal nets has not to be considered as relevant source of sporadic infections. The obtained data indicate that cryptosporidiosis is a disease of low epidemiological significance in Switzerland. To a great extent, cryptosporidiosis could be prevented by best known measures of personal hygiene, avoiding certain raw food-stuffs and being aware of safe catering on travels.

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

    PubMed

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

    2015-09-01

    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.

  14. 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.

  15. 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.

  16. 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... nationally tracked sources. Each licensee who manufactures a nationally tracked source after February 6,...

  17. 10 CFR 32.201 - Serialization of nationally tracked sources.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 10 Energy 1 2011-01-01 2011-01-01 false Serialization of nationally tracked sources. 32.201 Section 32.201 Energy NUCLEAR REGULATORY COMMISSION SPECIFIC DOMESTIC LICENSES TO MANUFACTURE OR TRANSFER... nationally tracked sources. Each licensee who manufactures a nationally tracked source after February 6,...

  18. 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.

  19. 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, S.K.K.; Morris, R.

    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.

  20. Cryptosporidium sp. Infections in Green Turtles, Chelonia mydas, as a Potential Source of Marine Waterborne Oocysts in the Hawaiian Islands

    PubMed Central

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

    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. PMID:16535658

  1. 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...

  2. Microbial Source Tracking in Adjacent Karst Springs

    PubMed Central

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

    2015-01-01

    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. PMID:26002893

  3. 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.

  4. 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. PMID:26422158

  5. 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.

  6. 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.

  7. 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

  8. 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.

  9. 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.

  10. 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...

  11. 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...

  12. 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.

  13. 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...

  14. 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

  15. 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. PMID:26301853

  16. Using RSS feeds to track open source radiology informatics projects

    NASA Astrophysics Data System (ADS)

    Nagy, Paul; Daly, Mark; Warnock, Michael; Siddiqui, Khan; Siegel, Eliot

    2005-04-01

    There are over 40 open source projects in the field of radiology informatics. Because these are organized and written by volunteers, the development speed varies greatly from one project to the next. To keep track of updates, users must constantly check in on each project's Web page. Many projects remain dormant for years, and ad hoc checking becomes both an inefficient and unreliable means of determining when new versions are available. The result is that most end users track only a few projects and are unaware when others that may be more germane to their interests leapfrog in development. RSS feeds provide a machine readable XML format to track software project updates. Currently only 8 of the 40 projects provide RSS feeds for automatic propagation of news updates. We have a built a news aggregation engine around open source projects in radiology informatics.

  17. Cryptosporidium muris in a Texas canine population.

    PubMed

    Lupo, Philip J; Langer-Curry, Rebecca C; Robinson, Mary; Okhuysen, Pablo C; Chappell, Cynthia L

    2008-06-01

    Molecular technology has led to the discovery of previously unrecognized Cryptosporidium species in new hosts, such as C. canis in humans. The notion that dogs may transmit Cryptosporidium species to humans has significant public health implications, and additional studies are merited. The purpose of this study was to examine a group of kenneled dogs to determine the prevalence of Cryptosporidium species infection and to identify parasite species. Prevalence of active infection was 71%. Six positive samples were analyzed by polymerase chain reaction amplification of the 18S ribosomal RNA gene followed by restriction fragment length polymorphism analysis to identify the Cryptosporidium species. Restriction digest patterns identified C. muris as the infecting species in all six dogs; species identity was confirmed by genetic sequencing. To our knowledge, this is the first report of a naturally occurring C. muris infection in a canine host. The finding of C. muris in asymptomatic canines supports the notion of dogs as potential sources of human infection.

  18. Tracking closely spaced multiple sources via spectral-estimation techniques

    NASA Astrophysics Data System (ADS)

    Gabriel, W. F.

    1982-06-01

    Modern spectral-estimation techniques have achieved a level of performance that attracts interest in applications area such as the tracking of multiple spatial sources. In addition to the original "superresolution' capability, these techniques offer an apparent 'absence of sidelobes' characteristic and some reasonable solutions to the difficult radar coherent-source problem that involves a phase-dependent SNR (signal-to-noise ratio) penalty. This report reviews the situation briefly, and it discusses a few of the techniques that have been found useful, including natural or synthetic doppler shifts, non-Toeplitz forward-backward subaperture-shift processing, and recent eigenvalue/eigenvector analysis algorithms. The techniques are applied to multiple-source situations that include mixtures of coherent and noncoherent sources of unequal strengths, with either an 8-or a 12-element linear-array sampling aperture. The first test case involves the estimation of six sources, two of which are 95% correlated. The second test case involves a tracking-simulation display example of four moving sources: three are -10dB coherent sources 95% correlated, and the other is a strong 20-dB noncoherent source. These test cases demonstrate the remarkable improvements obtained with the recent estimation techniques, and they point to the possibilities for real-world applications.

  19. 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.

  20. 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

  1. 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. PMID:26828187

  2. 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.

  3. 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

  4. COMPARISON OF RECURSIVE ESTIMATION TECHNIQUES FOR POSITION TRACKING RADIOACTIVE SOURCES

    SciTech Connect

    K. MUSKE; J. HOWSE

    2000-09-01

    This paper compares the performance of recursive state estimation techniques for tracking the physical location of a radioactive source within a room based on radiation measurements obtained from a series of detectors at fixed locations. Specifically, the extended Kalman filter, algebraic observer, and nonlinear least squares techniques are investigated. The results of this study indicate that recursive least squares estimation significantly outperforms the other techniques due to the severe model nonlinearity.

  5. Sound source tracking device for telematic spatial sound field reproduction

    NASA Astrophysics Data System (ADS)

    Cardenas, Bruno

    This research describes an algorithm that localizes sound sources for use in telematic applications. The localization algorithm is based on amplitude differences between various channels of a microphone array of directional shotgun microphones. The amplitude differences will be used to locate multiple performers and reproduce their voices, which were recorded at close distance with lavalier microphones, spatially corrected using a loudspeaker rendering system. In order to track multiple sound sources in parallel the information gained from the lavalier microphones will be utilized to estimate the signal-to-noise ratio between each performer and the concurrent performers.

  6. 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.

  7. 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/).

  8. Recursive Estimation for the Tracking of Radioactive Sources

    SciTech Connect

    Howse, J.W.; Muske, K.R.; Ticknor, L.O.

    1999-02-01

    This paper describes a recursive estimation algorithm used for tracking the physical location of radioactive sources in real-time as they are moved around in a facility. The al- gorithm is a nonlinear least squares estimation that mini- mizes the change in, the source location and the deviation between measurements and model predictions simultane- ously. The measurements used to estimate position consist of four count rates reported by four different gamma ray de tectors. There is an uncertainty in the source location due to the variance of the detected count rate. This work repre- sents part of a suite of tools which will partially automate security and safety assessments, allow some assessments to be done remotely, and provide additional sensor modalities with which to make assessments.

  9. Recursive estimation for the tracking of radioactive sources

    SciTech Connect

    Howse, J.W.; Ticknor, L.O.; Muske, K.R.

    1998-12-31

    This paper describes a recursive estimation algorithm used for tracking the physical location of radioactive sources in real-time as they are moved around in a facility. The algorithm is related to a nonlinear least squares estimation that minimizes the change in the source location and the deviation between measurements and model predictions simultaneously. The measurements used to estimate position consist of four count rates reported by four different gamma ray detectors. There is an uncertainty in the source location due to the large variance of the detected count rate. This work represents part of a suite of tools which will partially automate security and safety assessments, allow some assessments to be done remotely, and provide additional sensor modalities with which to make assessments.

  10. Tracking Nuclear Sources published by Well Servicing Magazine

    SciTech Connect

    Walker, Randy M

    2009-01-01

    Researchers at Oak Ridge National Laboratory are working with academia, the U.S. Government, and commercial companies to apply advanced secure Web 2.0 technologies to the tracking and monitoring of nuclear sources in the international supply chain. You ask, 'What is Web 2.0?' There is no rigid agreement on what Web 2.0 is, but Tim O'Reilly says it best when he defines Web 2.0 as 'the recent web innovations that have facilitated communication, information sharing, interoperability and collaboration.'

  11. [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.

  12. 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.

  13. 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.

  14. 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...

  15. 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...

  16. CRYPTOSPORIDIUM IN CATTLE FROM OBSERVING TO UNDERSTANDING

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryptosporidium parvum is a zoonotic pathogen transmissible from a variety of animals to humans and is a considerable public health concern. Dairy cattle have been identified in numerous reports as a major source of environmental contamination with this pathogen. However, virtually all reports have ...

  17. 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

  18. 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.

  19. 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 ...

  20. Tracking the Sources of Indium to the Environment

    NASA Astrophysics Data System (ADS)

    White, S. O.; Hemond, H.

    2009-12-01

    Indium is an important metal whose production is increasing dramatically due to new uses in the rapidly growing electronics, photovoltaic, and LED industries. Little is known about the natural or industrial cycling of indium, and although toxicity has been shown, toxicological data is incomplete and indium is not currently regulated in the United States. A review of the literature has shown that industrial releases of indium to the environment presently dominate natural fluxes. Mining and coal burning are the primary industrial sources of indium to the environment, with releases from the semiconductor industry small at present. This scenario may change with the rapid growth of semiconductor industries. In order to quantify releases of indium to the environment, it may be useful to exploit differences in the ratio of indium’s two stable isotopes, indium-113 and indium-115, from various sources. This method of source-tracking has proven useful for elements such as lead, where pollutant sources have significantly different isotopic ratios. Few measurements of indium isotopes have been published, and are mainly for purified indium metal or indium compounds; none are for environmental samples. The most recent measurements have shown 95.67 - 95.77% indium-115 and 4.23 - 4.33% indium-113, suggesting a range of natural isotopic abundances of at least 25 per mil. Our own measurements of highly purified, semiconductor grade indium show 95.9% indium-115 and 4.1% indium-113, indicating this range may be even larger. However, while the differences may be attributed to natural variation in isotopic ratios due to source differences or fractionation during processing, they could also be due to analytical error such as instrument fractionation. No isotopic reference material apparently exists with which to normalize measurements, therefore the reporting of absolute isotope percentages and lab-to-lab comparisons are difficult. Additionally, we have seen only a 0.5 per mil

  1. 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.

  2. 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

  3. Effects of drinking-water filtration on Cryptosporidium seroepidemiology, Scotland.

    PubMed

    Ramsay, Colin N; Wagner, Adam P; Robertson, Chris; Smith, Huw V; Pollock, Kevin G J

    2014-01-01

    Continuous exposure to low levels of Cryptosporidium oocysts is associated with production of protective antibodies. We investigated prevalence of antibodies against the 27-kDa Cryptosporidium oocyst antigen among blood donors in 2 areas of Scotland supplied by drinking water from different sources with different filtration standards: Glasgow (not filtered) and Dundee (filtered). During 2006-2009, seroprevalence and risk factor data were collected; this period includes 2007, when enhanced filtration was introduced to the Glasgow supply. A serologic response to the 27-kDa antigen was found for ≈75% of donors in the 2 cohorts combined. Mixed regression modeling indicated a 32% step-change reduction in seroprevalence of antibodies against Cryptosporidium among persons in the Glasgow area, which was associated with introduction of enhanced filtration treatment. Removal of Cryptosporidium oocysts from water reduces the risk for waterborne exposure, sporadic infections, and outbreaks. Paradoxically, however, oocyst removal might lower immunity and increase the risk for infection from other sources.

  4. 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.

  5. 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.

  6. 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...

  7. 10 CFR Appendix E to Part 20 - Nationally Tracked Source Thresholds

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 10 Energy 1 2012-01-01 2012-01-01 false Nationally Tracked Source Thresholds E Appendix E to Part 20 Energy NUCLEAR REGULATORY COMMISSION STANDARDS FOR PROTECTION AGAINST RADIATION Pt. 20, App. E Appendix E to Part 20— Nationally Tracked Source Thresholds The Terabecquerel (TBq) values are...

  8. 10 CFR Appendix E to Part 20 - Nationally Tracked Source Thresholds

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 10 Energy 1 2014-01-01 2014-01-01 false Nationally Tracked Source Thresholds E Appendix E to Part 20 Energy NUCLEAR REGULATORY COMMISSION STANDARDS FOR PROTECTION AGAINST RADIATION Pt. 20, App. E Appendix E to Part 20— Nationally Tracked Source Thresholds The Terabecquerel (TBq) values are...

  9. 10 CFR Appendix E to Part 20 - Nationally Tracked Source Thresholds

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 10 Energy 1 2011-01-01 2011-01-01 false Nationally Tracked Source Thresholds E Appendix E to Part 20 Energy NUCLEAR REGULATORY COMMISSION STANDARDS FOR PROTECTION AGAINST RADIATION Pt. 20, App. E Appendix E to Part 20— Nationally Tracked Source Thresholds The Terabecquerel (TBq) values are...

  10. 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

  11. 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

  12. Source tracking Mycobacterium ulcerans infections in the Ashanti region, Ghana.

    PubMed

    Narh, Charles A; Mosi, Lydia; Quaye, Charles; Dassi, Christelle; Konan, Daniele O; Tay, Samuel C K; de Souza, Dziedzom K; Boakye, Daniel A; Bonfoh, Bassirou

    2015-01-01

    Although several studies have associated Mycobacterium ulcerans (MU) infection, Buruli ulcer (BU), with slow moving water bodies, there is still no definite mode of transmission. Ecological and transmission studies suggest Variable Number Tandem Repeat (VNTR) typing as a useful tool to differentiate MU strains from other Mycolactone Producing Mycobacteria (MPM). Deciphering the genetic relatedness of clinical and environmental isolates is seminal to determining reservoirs, vectors and transmission routes. In this study, we attempted to source-track MU infections to specific water bodies by matching VNTR profiles of MU in human samples to those in the environment. Environmental samples were collected from 10 water bodies in four BU endemic communities in the Ashanti region, Ghana. Four VNTR loci in MU Agy99 genome, were used to genotype environmental MU ecovars, and those from 14 confirmed BU patients within the same study area. Length polymorphism was confirmed with sequencing. MU was present in the 3 different types of water bodies, but significantly higher in biofilm samples. Four MU genotypes, designated W, X, Y and Z, were typed in both human and environmental samples. Other reported genotypes were only found in water bodies. Animal trapping identified 1 mouse with lesion characteristic of BU, which was confirmed as MU infection. Our findings suggest that patients may have been infected from community associated water bodies. Further, we present evidence that small mammals within endemic communities could be susceptible to MU infections. M. ulcerans transmission could involve several routes where humans have contact with risk environments, which may be further compounded by water bodies acting as vehicles for disseminating strains.

  13. Microbial source tracking in a coastal California watershed reveals canines as controllable sources of fecal contamination.

    PubMed

    Ervin, Jared S; Van De Werfhorst, Laurie C; Murray, Jill L S; Holden, Patricia A

    2014-08-19

    Elevated levels of fecal indicator bacteria (FIB), including Escherichia coli and enterococci, trigger coastal beach advisories and signal public health risks. Solving FIB pollution in suburban coastal watersheds is challenging, as there are many potential sources. The Arroyo Burro watershed in Santa Barbara, CA is an example, with its popular, but chronically FIB-contaminated beach. To address, a microbial source tracking study was performed. Surface waters were sampled over 2 years, FIB were quantified, and DNA was analyzed for host-associated fecal markers. Surf zone FIB were only elevated when the coastal lagoon was discharging. Among the fecal sources into the lagoon, including upstream human sources and coastal birds, canines were the most important. Canine sources included input via upstream creek water, which decreased after creek-side residences were educated about proper pet waste disposal, and direct inputs to the lagoon and surf zone, where dog waste could have been tidally exchanged with the lagoon. Based on this study, canine waste can be an influential, yet controllable, fecal source to suburban coastal beaches.

  14. 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).

  15. 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.

  16. 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.

  17. MOLECULAR DETECTION OF CRYPTOSPORIDIUM OOCYSTS IN WATER: THE CHALLENGE AND PROMISE

    EPA Science Inventory

    Because of the presence of host-adapted Cryptosporidium species and genotypes, molecular tools can help assess the source and hazardous potential of Cryptosporidium oocysts in water. The development and use of molecular tools in the analysis of environmental samples have gone tho...

  18. Inactivation kinetics of Cryptosporidium parvum oocysts in swine waste lagoon and spray field

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Because of outbreaks of cryptosporidiosis in humans, Cryptosporidium has become a public health concern. Commercial swine operations can be a source of this protozoan parasite. Although the species distribution of Cryptosporidium is likely dominated by C. suis, a fraction may be comprised of the zoo...

  19. Human cryptosporidiosis caused by Cryptosporidium tyzzeri and C. parvum isolates presumably transmitted from wild mice.

    PubMed

    Rasková, Veronika; Kvetonová, Dana; Sak, Bohumil; McEvoy, John; Edwinson, Adam; Stenger, Brianna; Kvác, Martin

    2013-01-01

    We report a case of severe human cryptosporidiosis caused by Cryptosporidium tyzzeri and C. parvum with an unusually high frequency of liquid stools. Wild mice were the most likely source of infection, demonstrating the potential for wild-mouse-borne Cryptosporidium to infect humans and highlighting the health risks associated with synantropic rodents.

  20. Cryptosporidium tyzzeri and Cryptosporidium pestis: which name is valid?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The validity of the name Cryptosporidium tyzzeri has been questioned because this name was previously used for a Cryptosporidium species in chickens in the original description by E. E. Tyzzer in 1929 which was later given the name by N.D. Levine in 1961. To further complicate matters, this specie...

  1. Sourcing sediment loss to watercourses at catchment scale using a novel tracing-tracking framework

    NASA Astrophysics Data System (ADS)

    Collins, Adrian; Zhang, Yusheng; Walling, Des; Black, Kevin

    2010-05-01

    Although traditional sediment tracing approaches provide valuable information for characterising key generic sediment sources, Catchment Officers working as part of the England Catchment Sensitive Farming Delivery Initiative (ECSFDI) frequently require higher resolution evidence to assist better the targeting of mitigation options. Accordingly, a novel framework combining conventional sediment source fingerprinting and a dual signature tracking method has recently been used to improve the resolution of sediment source information for contrasting priority catchments. Conventional geochemical tracing incorporating revised mass balance modelling is used to provide information on the relative significance of generic sediment sources such as grassland or arable surface soils, damaged road verges and channel banks/subsurface sources and to provide a framework for the spatial extrapolation of tracking data. Particle tracking using fluorescent-magnetic grains is used to elucidate sediment loss from key components of the primary generic sources, characterised, for example, as poached gateways or cattle tracks and wider areas of general poaching damage in grass fields, wheeling or inter-wheeling areas in arable fields and poached or fluvial-eroded channel banks. The insertion of high-strength magnets in watercourses ensures that the tracking component links sediment loss from seeded areas to river channels as opposed to providing edge-of-field information. Uncertainty and prior information are explicitly recognised by the novel framework. Key words: sediment sources; catchment scale; fingerprinting; tracking; uncertainty; prior information

  2. 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.

  3. 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.

  4. 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

  5. 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...

  6. 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...

  7. 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.

  8. 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.

  9. Implications of biofilm-associated waterborne Cryptosporidium oocysts for the water industry.

    PubMed

    Angles, Mark L; Chandy, Joseph P; Cox, Peter T; Fisher, Ian H; Warnecke, Malcolm R

    2007-08-01

    Waterborne Cryptosporidium has been responsible for drinking water-associated disease outbreaks in a number of developed countries. As a result of the resistance of Cryptosporidium to chlorine, which is typically applied as a final barrier to protect the quality of distributed drinking water, current management practices are focused on source-water management and water treatment as ways of preventing Cryptosporidium from entering drinking-water supplies. In the event that treatment barriers fail, surprisingly little is known of the fate of oocysts once they enter a distribution system. To assess properly the risks of waterborne Cryptosporidium, a more thorough understanding of the fate of oocysts in water distribution systems, with emphasis on Cryptosporidium-biofilm interactions, is required.

  10. Spatiotemporal Analysis of Cryptosporidium Species/Genotypes and Relationships with Other Zoonotic Pathogens in Surface Water from Mixed-Use Watersheds

    PubMed Central

    Wilkes, Graham; Ruecker, Norma J.; Neumann, Norman F.; Gannon, Victor P. J.; Jokinen, Cassandra; Sunohara, Mark; Topp, Edward; Pintar, Katarina D. M.; Edge, Thomas A.

    2013-01-01

    Nearly 690 raw surface water samples were collected during a 6-year period from multiple watersheds in the South Nation River basin, Ontario, Canada. Cryptosporidium oocysts in water samples were enumerated, sequenced, and genotyped by detailed phylogenetic analysis. The resulting species and genotypes were assigned to broad, known host and human infection risk classes. Wildlife/unknown, livestock, avian, and human host classes occurred in 21, 13, 3, and <1% of sampled surface waters, respectively. Cryptosporidium andersoni was the most commonly detected livestock species, while muskrat I and II genotypes were the most dominant wildlife genotypes. The presence of Giardia spp., Salmonella spp., Campylobacter spp., and Escherichia coli O157:H7 was evaluated in all water samples. The greatest significant odds ratios (odds of pathogen presence when host class is present/odds of pathogen presence when host class is absent) for Giardia spp., Campylobacter spp., and Salmonella spp. in water were associated, respectively, with livestock (odds ratio of 3.1), avian (4.3), and livestock (9.3) host classes. Classification and regression tree analyses (CART) were used to group generalized host and human infection risk classes on the basis of a broad range of environmental and land use variables while tracking cooccurrence of zoonotic pathogens in these groupings. The occurrence of livestock-associated Cryptosporidium was most strongly related to agricultural water pollution in the fall (conditions also associated with elevated odds ratios of other zoonotic pathogens occurring in water in relation to all sampling conditions), whereas wildlife/unknown sources of Cryptosporidium were geospatially associated with smaller watercourses where urban/rural development was relatively lower. Conditions that support wildlife may not necessarily increase overall human infection risks associated with Cryptosporidium since most Cryptosporidium genotypes classed as wildlife in this study (e

  11. 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 ...

  12. LIBRARY-DEPENDENT MICROBIAL SOURCE TRACKING OF ENTEROCOCCUS SP. USING AFLP AND BOX-PCR

    EPA Science Inventory

    Library-dependent microbial source tracking (LD MST) methods are one of the approaches used to identify nonpoint sources of fecal contamination in support of total maximum daily load implementation. However, LD MST methods have been questioned due to the high temporal and spatial...

  13. QUO VADIS SOURCE TRACKING? TOWARDS A STRATEGIC FRAMEWORK FOR ENVIRONMENTAL MONITORING OF FECAL POLLUTION

    EPA Science Inventory

    Advances in microbial source tracking (MST) have largely been driven by the need to comply with water quality standards based on traditional indicator bacteria. Recently, a number of PCR-based, culture- and library-independent methods have been gaining popularity among source tra...

  14. 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...

  15. 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. PMID:25769247

  16. 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. PMID:25988830

  17. 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.

  18. An efficient central DOA tracking algorithm for multiple incoherently distributed sources

    NASA Astrophysics Data System (ADS)

    Hassen, Sonia Ben; Samet, Abdelaziz

    2015-12-01

    In this paper, we develop a new tracking method for the direction of arrival (DOA) parameters assuming multiple incoherently distributed (ID) sources. The new approach is based on a simple covariance fitting optimization technique exploiting the central and noncentral moments of the source angular power densities to estimate the central DOAs. The current estimates are treated as measurements provided to the Kalman filter that model the dynamic property of directional changes for the moving sources. Then, the covariance-fitting-based algorithm and the Kalman filtering theory are combined to formulate an adaptive tracking algorithm. Our algorithm is compared to the fast approximated power iteration-total least square-estimation of signal parameters via rotational invariance technique (FAPI-TLS-ESPRIT) algorithm using the TLS-ESPRIT method and the subspace updating via FAPI-algorithm. It will be shown that the proposed algorithm offers an excellent DOA tracking performance and outperforms the FAPI-TLS-ESPRIT method especially at low signal-to-noise ratio (SNR) values. Moreover, the performances of the two methods increase as the SNR values increase. This increase is more prominent with the FAPI-TLS-ESPRIT method. However, their performances degrade when the number of sources increases. It will be also proved that our method depends on the form of the angular distribution function when tracking the central DOAs. Finally, it will be shown that the more the sources are spaced, the more the proposed method can exactly track the DOAs.

  19. 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.

  20. 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

  1. 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

  2. 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

  3. Cryptosporidium Genotypes in Wildlife from a New York Watershed▿

    PubMed Central

    Feng, Yaoyu; Alderisio, Kerri A.; Yang, Wenli; Blancero, Lisa A.; Kuhne, William G.; Nadareski, Christopher A.; Reid, Michael; Xiao, Lihua

    2007-01-01

    To identify the animal sources for Cryptosporidium contamination, we genotyped Cryptosporidium spp. in wildlife from the watershed of the New York City drinking water supply, using a small-subunit rRNA gene-based PCR-restriction fragment length polymorphism analysis and DNA sequencing. A total of 541 specimens from 38 species of wildlife were analyzed. One hundred and eleven (20.5%) of the wildlife specimens were PCR positive. Altogether, 21 Cryptosporidium genotypes were found in wildlife samples, 11 of which were previously found in storm runoff in the watershed, and six of these 11 were from storm water genotypes of unknown animal origin. Four new genotypes were found, and the animal hosts for four storm water genotypes were expanded. With the exception of the cervine genotype, most genotypes were found in a limited number of animal species and have no major public health significance. PMID:17720824

  4. Cryptosporidium genotypes in wildlife from a new york watershed.

    PubMed

    Feng, Yaoyu; Alderisio, Kerri A; Yang, Wenli; Blancero, Lisa A; Kuhne, William G; Nadareski, Christopher A; Reid, Michael; Xiao, Lihua

    2007-10-01

    To identify the animal sources for Cryptosporidium contamination, we genotyped Cryptosporidium spp. in wildlife from the watershed of the New York City drinking water supply, using a small-subunit rRNA gene-based PCR-restriction fragment length polymorphism analysis and DNA sequencing. A total of 541 specimens from 38 species of wildlife were analyzed. One hundred and eleven (20.5%) of the wildlife specimens were PCR positive. Altogether, 21 Cryptosporidium genotypes were found in wildlife samples, 11 of which were previously found in storm runoff in the watershed, and six of these 11 were from storm water genotypes of unknown animal origin. Four new genotypes were found, and the animal hosts for four storm water genotypes were expanded. With the exception of the cervine genotype, most genotypes were found in a limited number of animal species and have no major public health significance.

  5. 40 CFR 141.713 - Schedule for compliance with Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... the bin classification for a filtered system changes following the second round of source water... AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced... Cryptosporidium treatment requirements. (a) Following initial bin classification under § 141.710(c),...

  6. 40 CFR 141.713 - Schedule for compliance with Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... the bin classification for a filtered system changes following the second round of source water... AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced... Cryptosporidium treatment requirements. (a) Following initial bin classification under § 141.710(c),...

  7. 40 CFR 141.713 - Schedule for compliance with Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... the bin classification for a filtered system changes following the second round of source water... AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced... Cryptosporidium treatment requirements. (a) Following initial bin classification under § 141.710(c),...

  8. 40 CFR 141.713 - Schedule for compliance with Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... the bin classification for a filtered system changes following the second round of source water... AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced... Cryptosporidium treatment requirements. (a) Following initial bin classification under § 141.710(c),...

  9. 40 CFR 141.713 - Schedule for compliance with Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... the bin classification for a filtered system changes following the second round of source water... AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced... Cryptosporidium treatment requirements. (a) Following initial bin classification under § 141.710(c),...

  10. 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...

  11. Dipole source encoding and tracking by the goldfish auditory system

    PubMed Central

    Coombs, Sheryl; Fay, Richard R.; Elepfandt, Andreas

    2010-01-01

    In goldfish and other otophysans, the Weberian ossicles mechanically link the saccule of the inner ear to the anterior swimbladder chamber (ASB). These structures are correlated with enhanced sound-pressure sensitivity and greater sensitivity at high frequencies (600–2000 Hz). However, surprisingly little is known about the potential impact of the ASB on other otolithic organs and about how auditory responses are modulated by discrete sources that change their location or orientation with respect to the ASB. In this study, saccular and lagenar nerve fiber responses and conditioned behaviors of goldfish were measured to a small, low-frequency (50 Hz) vibrating sphere (dipole) source as a function of its location along the body and its orientation with respect to the ASB. Conditioned behaviors and saccular nerve fiber activity exhibited response characteristics nearly identical to those measured from a hydrophone in the same relative position as the ASB. By contrast, response patterns from lagena fibers could not be predicted by pressure inputs to the ASB. Deflation of the ASB abolished the characteristic spatial response pattern of saccular but not lagena fibers. These results show that: (1) the lagena is not driven by ASB-mediated pressure inputs to the ear; (2) the ASB–saccule pathway dominates behavioral responsiveness, operating effectively at frequencies as low as 50 Hz; and (3) behavioral and neural (saccular) responses are strongly modulated by the position and orientation of the dipole with respect to the ASB. PMID:20889834

  12. 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.

  13. Prevalence of Cryptosporidium species among HIV positive asymptomatic and symptomatic immigrant population in Kashmir, India

    PubMed Central

    Masarat, S; Ahmad, F; Chisti, M; Hamid, S; Sofi, B Ahmad

    2012-01-01

    Background and Objectives Cryptosporidiosis has not been reported as an endemic disease in Kashmir, but high prevalence of Cryptosporidium sp. has been found among asymptomatic (non-diarrheic) HIV positive immigrants in present study. Due to increasing number of HIV positive immigrants in Kashmir, Cryptosporidium may become a public health problem in Kashmir. Materials and Methods A total of 45 stool samples were obtained from symptomatic (diarrheic n = 9) and asymptomatic (non-diarrheic n = 36) patients infected with HIV. The stool samples were concentrated using formalin ethyl acetate concentration technique, stained with modified Kinyoun's cold stain and oocysts were identified by microscopy under 1000 x magnification. It was confirmed by detection of antigens in stool samples by ELISA. Results It was established that all the patients studied were carriers of Cryptosporidium. In present study though 80% of patients were asymptomatic (non-diarrheic) and HIV positive which involved non-Kashmiri army personals and travelers (immigrants) but were carriers of Cryptosporidium and 20% of HIV positive patients were emigrants (local Kashmiri traders) who travelled different states of India were having diarrhea (symptomatic) as well as carrier of Cryptosporidium. Conclusion Though Cryptosporidium infection causes chronic diarrhea but in present study all HIV positive patients screened whether diarrheic or non-diarrheic were positive for Cryptosporidium. To prevent the transmission of Cryptosporidium oocyst in environment and endemic spread of cryptosporidiosis as non-diarrheic HIV positive population may be potential source of infection, obligatory laboratory testing for Cryptosporidium in HIV positive immigrant population like traders and travelers is highly recommended in order to have a better understanding of the cause of spread Cryptosporidium infection in Kashmir. PMID:22783459

  14. 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...

  15. 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...

  16. 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...

  17. 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

  18. 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 ...

  19. 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...

  20. 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...

  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. 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. PMID:20572154

  3. 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

  4. 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

  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. 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.

  7. Patterns of Cryptosporidium Oocyst Shedding by Eastern Grey Kangaroos Inhabiting an Australian Watershed

    PubMed Central

    Power, Michelle L.; Sangster, Nicholas C.; Slade, Martin B.; Veal, Duncan A.

    2005-01-01

    The occurrence of Cryptosporidium oocysts in feces from a population of wild eastern grey kangaroos inhabiting a protected watershed in Sydney, Australia, was investigated. Over a 2-year period, Cryptosporidium oocysts were detected in 239 of the 3,557 (6.7%) eastern grey kangaroo fecal samples tested by using a combined immunomagnetic separation and flow cytometric technique. The prevalence of Cryptosporidium in this host population was estimated to range from 0.32% to 28.5%, with peaks occurring during the autumn months. Oocyst shedding intensity ranged from below 20 oocysts/g feces to 2.0 × 106 oocysts/g feces, and shedding did not appear to be associated with diarrhea. Although morphologically similar to the human-infective Cryptosporidium hominis and the Cryptosporidium parvum “bovine” genotype oocysts, the oocysts isolated from kangaroo feces were identified as the Cryptosporidium “marsupial” genotype I or “marsupial” genotype II. Kangaroos are the predominant large mammal inhabiting Australian watersheds and are potentially a significant source of Cryptosporidium contamination of drinking water reservoirs. However, this host population was predominantly shedding the marsupial-derived genotypes, which to date have been identified only in marsupial host species. PMID:16204534

  8. 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.

  9. 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. PMID:26797442

  10. Patterns of Cryptosporidium oocyst shedding by eastern grey kangaroos inhabiting an Australian watershed.

    PubMed

    Power, Michelle L; Sangster, Nicholas C; Slade, Martin B; Veal, Duncan A

    2005-10-01

    The occurrence of Cryptosporidium oocysts in feces from a population of wild eastern grey kangaroos inhabiting a protected watershed in Sydney, Australia, was investigated. Over a 2-year period, Cryptosporidium oocysts were detected in 239 of the 3,557 (6.7%) eastern grey kangaroo fecal samples tested by using a combined immunomagnetic separation and flow cytometric technique. The prevalence of Cryptosporidium in this host population was estimated to range from 0.32% to 28.5%, with peaks occurring during the autumn months. Oocyst shedding intensity ranged from below 20 oocysts/g feces to 2.0 x 10(6) oocysts/g feces, and shedding did not appear to be associated with diarrhea. Although morphologically similar to the human-infective Cryptosporidium hominis and the Cryptosporidium parvum "bovine" genotype oocysts, the oocysts isolated from kangaroo feces were identified as the Cryptosporidium "marsupial" genotype I or "marsupial" genotype II. Kangaroos are the predominant large mammal inhabiting Australian watersheds and are potentially a significant source of Cryptosporidium contamination of drinking water reservoirs. However, this host population was predominantly shedding the marsupial-derived genotypes, which to date have been identified only in marsupial host species.

  11. An outbreak of Cryptosporidium at a recreational water park in Niagara Region, Canada.

    PubMed

    Hopkins, Jessica; Hague, Heather; Hudgin, Glen; Ross, Lorrie; Moore, Deborah

    2013-05-01

    Cryptosporidium is a parasitic protozoan found in water sources and spread through the fecal-oral route. Cryptosporidiosis is characterized by gastroenteritis and is increasingly associated with recreational water sources. On December 3, 2010, Niagara Region Public Health was informed of a laboratory-confirmed case of Cryptosporidium. Over the subsequent two weeks, a total of three additional laboratory-confirmed cases were reported. All cases had visited the same water park in Niagara Region, Canada, over November 14-16, 2010. A total of 12 cases associated with the outbreak ranged in age from 1 to 66 years. This article describes the outbreak, environmental investigation, and control measures. The environmental investigation revealed that the ultraviolet disinfection system was offline on November 14, 2010, which may have allowed for the transmission of Cryptosporidium to bathers. Further research into the detection of Cryptosporidium outbreaks and regulations and guidelines for water park operators may help to decrease future outbreaks.

  12. Occurrence of Giardia and Cryptosporidium spp. in surface water supplies.

    PubMed Central

    LeChevallier, M W; Norton, W D; Lee, R G

    1991-01-01

    Giardia and Cryptosporidium levels were determined by using a combined immunofluorescence test for source waters of 66 surface water treatment plants in 14 states and 1 Canadian province. The results showed that cysts and oocysts were widely dispersed in the aquatic environment. Giardia spp. were detected in 81% of the raw water samples. Cryptosporidium spp. were found in 87% of the raw water locations. Overall, Giardia or Cryptosporidium spp. were detected in 97% of the raw water samples. Higher cyst and oocyst densities were associated with source waters receiving industrial or sewage effluents. Significant correlations were found between Giardia and Cryptosporidium densities and raw water quality parameters such as turbidity and total and fecal coliform levels. Statistical modeling suggests that cyst and oocyst densities could be predicted on the basis of watershed and water quality characteristics. The occurrence of high levels of Giardia cysts in raw water samples may require water utilities to apply treatment beyond that outlined in the Surface Water Treatment Rule of the U.S. Environmental Protection Agency. PMID:1822675

  13. Enhancing fecal coliform total maximum daily load models through bacterial source tracking

    USGS Publications Warehouse

    Hyer, K.E.; Moyer, D.L.

    2004-01-01

    Surface water impairment by fecal coliform bacteria is a water quality issue of national scope and importance. In Virginia, more than 400 stream and river segments are on the Commonwealth's 2002 303(d) list because of fecal coliform impairment. Total maximum daily loads (TMDLs) will be developed for most of these listed streams and rivers. Information regarding the major fecal coliform sources that impair surface water quality would enhance the development of effective watershed models and improve TMDLs. Bacterial source tracking (BST) is a recently developed technology for identifying the sources of fecal coliform bacteria and it may be helpful in generating improved TMDLs. Bacterial source tracking was performed, watershed models were developed, and TMDLs were prepared for three streams (Accotink Creek, Christians Creek, and Blacks Run) on Virginia's 303(d) list of impaired waters. Quality assurance of the BST work suggests that these data adequately describe the bacteria sources that are impairing these streams. Initial comparison of simulated bacterial sources with the observed BST data indicated that the fecal coliform sources were represented inaccurately in the initial model simulation. Revised model simulations (based on BST data) appeared to provide a better representation of the sources of fecal coliform bacteria in these three streams. The coupled approach of incorporating BST data into the fecal coliform transport model appears to reduce model uncertainty and should result in an improved TMDL.

  14. 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.

  15. Rapid identification and source-tracking of Listeria monocytogenes using MALDI-TOF mass spectrometry.

    PubMed

    Jadhav, Snehal; Gulati, Vandana; Fox, Edward M; Karpe, Avinash; Beale, David J; Sevior, Danielle; Bhave, Mrinal; Palombo, Enzo A

    2015-06-01

    Listeria monocytogenes is an important foodborne pathogen responsible for the sometimes fatal disease listeriosis. Public health concerns and stringent regulations associated with the presence of this pathogen in food and food processing environments underline the need for rapid and reliable detection and subtyping techniques. In the current study, the application of matrix assisted laser desorption/ionisation-time-of-flight mass spectrometry (MALDI-TOF MS) as a single identification and source-tracking tool for a collection of L. monocytogenes isolates, obtained predominantly from dairy sources within Australia, was explored. The isolates were cultured on different growth media and analysed using MALDI-TOF MS at two incubation times (24 and 48 h). Whilst reliable genus-level identification was achieved from most media, identification at the species level was found to be dependent on culture conditions. Successful speciation was highest for isolates cultured on the chromogenic Agar Listeria Ottaviani Agosti agar (ALOA, 91% of isolates) and non-selective horse blood agar (HBA, 89%) for 24h. Chemometric statistical analysis of the MALDI-TOF MS data enabled source-tracking of L. monocytogenes isolates obtained from four different dairy sources. Strain-level discrimination was also observed to be influenced by culture conditions. In addition, t-test/analysis of variance (ANOVA) was used to identify potential biomarker peaks that differentiated the isolates according to their source of isolation. Source-tracking using MALDI-TOF MS was compared and correlated with the gold standard pulsed-field gel electrophoresis (PFGE) technique. The discriminatory index and the congruence between both techniques were compared using the Simpsons Diversity Index and adjusted Rand and Wallace coefficients. Overall, MALDI-TOF MS based source-tracking (using data obtained by culturing the isolates on HBA) and PFGE demonstrated good congruence with a Wallace coefficient of 0.71 and

  16. 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.

  17. 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.

  18. Current Tracking Control of Voltage Source PWM Inverters Using Adaptive Digital Signal Processing

    NASA Astrophysics Data System (ADS)

    Fukuda, Shoji; Furukawa, Yuya

    An active filter (AF) is required to have a high control capability of tracking a time-varying current reference. However, a steady-state current error always exists if a conventional proportional and integral (PI) regulator is used because the current reference varies in time. This paper proposes the application of adaptive digital signal processing (ADSP) to the current control of voltage source PWM inverters. ADSP does not require any additional hardware. It can automatically minimize the mean square-error. Since the processing time available by a computer is limited, ADSP cannot eliminate higher order harmonics but can eliminate lower order harmonics such as 5th to 17th. Experimental results demonstrate that ADSP is useful for improving the reference tracking performance of voltage source inverters.

  19. Tracking brachytherapy sources using emission imaging with one flat panel detector

    SciTech Connect

    Song Haijun; Bowsher, James; Das, Shiva; Yin Fangfang

    2009-04-15

    This work proposes to use the radiation from brachytherapy sources to track their dwell positions in three-dimensional (3D) space. The prototype device uses a single flat panel detector and a BB tray. The BBs are arranged in a defined pattern. The shadow of the BBs on the flat panel is analyzed to derive the 3D coordinates of the illumination source, i.e., the dwell position of the brachytherapy source. A kilovoltage x-ray source located 3.3 m away was used to align the center BB with the center pixel on the flat panel detector. For a test plan of 11 dwell positions, with an Ir-192 high dose rate unit, one projection was taken for each dwell point, and locations of the BB shadows were manually identified on the projection images. The 3D coordinates for the 11 dwell positions were reconstructed based on two BBs. The distances between dwell points were compared with the expected values. The average difference was 0.07 cm with a standard deviation of 0.15 cm. With automated BB shadow recognition in the future, this technique possesses the potential of tracking the 3D trajectory and the dwell times of a brachytherapy source in real time, enabling real time source position verification.

  20. 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.

  1. Occurrence of Cryptosporidium oocysts and Giardia cysts in water supplies of San Pedro Sula, Honduras.

    PubMed

    Solo-Gabriele, H M; LeRoy Ager, A; Fitzgerald Lindo, J; Dubón, J M; Neumeister, S M; Baum, M K; Palmer, C J

    1998-12-01

    During June 1996, water supplies of the city of San Pedro Sula, Honduras, were sampled to obtain an assessment of Cryptosporidium oocyst and Giardia cyst concentrations. Each sample was concentrated and stained with an indirect immunofluorescent antibody, and parasites were counted through microscopic analysis. In three surface water supplies, Cryptosporidium oocyst concentrations ranged from 58 to 260 oocysts per 100 L, and Giardia cysts were present in concentrations ranging from 380 to 2100 cysts per 100 L. Unlike the surface water samples, groundwater had a higher concentration of Cryptosporidium oocysts (26/100 L) than Giardia cysts (6/100 L), suggesting that the groundwater aquifer protects the water supply more effectively from larger Giardia cysts. Cryptosporidium oocyst concentrations are within the typical range for surface water supplies in North America whereas Giardia cyst concentrations are elevated. Efforts should be made to protect raw water from sources of contamination.

  2. 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...

  3. Source tracking of leaky sewers: a novel approach combining fecal indicators in water and sediments.

    PubMed

    Guérineau, Hélène; Dorner, Sarah; Carrière, Annie; McQuaid, Natasha; Sauvé, Sébastien; Aboulfadl, Khadija; Hajj-Mohamad, Mariam; Prévost, Michèle

    2014-07-01

    In highly urbanized areas, surface water and groundwater are particularly vulnerable to sewer exfiltration. In this study, as an alternative to Microbial Source Tracking (MST) methods, we propose a new method combining microbial and chemical fecal indicators (Escherichia coli (E. coli)) and wastewater micropollutants (WWMPs) analysis both in water and sediment samples and under different meteorological conditions. To illustrate the use of this method, wastewater exfiltration and subsequent infiltration were identified and quantified by a three-year field study in an urban canal. The gradients of concentrations observed suggest that several sources of fecal contamination of varying intensity may be present along the canal, including feces from resident animal populations, contaminated surface run-off along the banks and under bridge crossings, release from contaminated banks, entrainment of contaminated sediments, and most importantly sewage exfiltration. Calculated exfiltration-infiltration volumes varied between 0.6 and 15.7 m(3)/d per kilometer during dry weather, and between 1.1 and 19.5 m(3)/d per kilometer during wet weather. WWMPs were mainly diluted and degraded below detection limits in water. E. coli remains the best exfiltration indicator given a large volume of dilution and a high abundance in the wastewater source. WWMPs are effective for detecting cumulated contamination in sediments from a small volume source and are particularly important because E. coli on its own does not allow source tracking. PMID:24735912

  4. Source tracking of leaky sewers: a novel approach combining fecal indicators in water and sediments.

    PubMed

    Guérineau, Hélène; Dorner, Sarah; Carrière, Annie; McQuaid, Natasha; Sauvé, Sébastien; Aboulfadl, Khadija; Hajj-Mohamad, Mariam; Prévost, Michèle

    2014-07-01

    In highly urbanized areas, surface water and groundwater are particularly vulnerable to sewer exfiltration. In this study, as an alternative to Microbial Source Tracking (MST) methods, we propose a new method combining microbial and chemical fecal indicators (Escherichia coli (E. coli)) and wastewater micropollutants (WWMPs) analysis both in water and sediment samples and under different meteorological conditions. To illustrate the use of this method, wastewater exfiltration and subsequent infiltration were identified and quantified by a three-year field study in an urban canal. The gradients of concentrations observed suggest that several sources of fecal contamination of varying intensity may be present along the canal, including feces from resident animal populations, contaminated surface run-off along the banks and under bridge crossings, release from contaminated banks, entrainment of contaminated sediments, and most importantly sewage exfiltration. Calculated exfiltration-infiltration volumes varied between 0.6 and 15.7 m(3)/d per kilometer during dry weather, and between 1.1 and 19.5 m(3)/d per kilometer during wet weather. WWMPs were mainly diluted and degraded below detection limits in water. E. coli remains the best exfiltration indicator given a large volume of dilution and a high abundance in the wastewater source. WWMPs are effective for detecting cumulated contamination in sediments from a small volume source and are particularly important because E. coli on its own does not allow source tracking.

  5. Source tracking aerosols released from land-applied class B biosolids during high-wind events.

    PubMed

    Baertsch, Carolina; Paez-Rubio, Tania; Viau, Emily; Peccia, Jordan

    2007-07-01

    DNA-based microbial source tracking (MST) methods were developed and used to specifically and sensitively track the unintended aerosolization of land-applied, anaerobically digested sewage sludge (biosolids) during high-wind events. Culture and phylogenetic analyses of bulk biosolids provided a basis for the development of three different MST methods. They included (i) culture- and 16S rRNA gene-based identification of Clostridium bifermentans, (ii) direct PCR amplification and sequencing of the 16S rRNA gene for an uncultured bacterium of the class Chloroflexi that is commonly present in anaerobically digested biosolids, and (iii) direct PCR amplification of a 16S rRNA gene of the phylum Euryarchaeota coupled with terminal restriction fragment length polymorphism to distinguish terminal fragments that are unique to biosolid-specific microorganisms. Each method was first validated with a broad group of bulk biosolids and soil samples to confirm the target's exclusive presence in biosolids and absence in soils. Positive responses were observed in 100% of bulk biosolid samples and in less than 11% of the bulk soils tested. Next, a sampling campaign was conducted in which all three methods were applied to aerosol samples taken upwind and downwind of fields that had recently been land applied with biosolids. When average wind speeds were greater than 5 m/s, source tracking results confirmed the presence of biosolids in 56% of the downwind samples versus 3% of the upwind samples. During these high-wind events, the biosolid concentration in downwind aerosols was between 0.1 and 2 microg/m3. The application of DNA-based source tracking to aerosol samples has confirmed that wind is a possible mechanism for the aerosolization and off-site transport of land-applied biosolids. PMID:17513591

  6. CRYPTOSPORIDIUM INACTIVATION AND REMOVAL RESEARCH

    EPA Science Inventory

    Bench- and pilot-scale tests were performed to assess the ability of conventional treatment, ozonation and chlorine dioxide to remove and inactivate Cryptosporidium oocysts. The impacts of coagulant type, coagulant dose, raw water quality, filter loading rates and filter media w...

  7. Human cryptosporidiosis diagnosed in Western Australia: a mixed infection with Cryptosporidium meleagridis, the Cryptosporidium mink genotype, and an unknown Cryptosporidium species.

    PubMed

    Ng-Hublin, Josephine S Y; Combs, Barry; Mackenzie, Brian; Ryan, Una

    2013-07-01

    This report describes a case of cryptosporidiosis from an immunocompetent patient from Perth, Western Australia, suffering from diarrhea and a spectrum of other symptoms. Molecular identification revealed that this patient was infected with three Cryptosporidium species-Cryptosporidium meleagridis, the Cryptosporidium mink genotype, and an unknown Cryptosporidium species.

  8. 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. ...

  9. Tracking magmatic intrusions in real-time by means of free-shaped volcanic source modelling

    NASA Astrophysics Data System (ADS)

    Cannavo', Flavio; Camacho, Antonio G.; Scandura, Danila; González, Pablo J.; Mattia, Mario; Fernández, José

    2014-05-01

    Nowadays continuous measurements of geophysical parameters provide a general real-time view of current state of the volcano. Nonetheless, a current challenge is to localize and track in real-time the evolution of the magma source beneath the volcano. Here we present a new methodology to rapidly estimate magmatic sources from surface geodetic data and track their evolution in time without any a priori assumption about source geometry. Indeed, the proposed approach takes the advantages of fast calculation from the analytical models and adds the capability to model free-shape distributed sources. Assuming homogenous elastic conditions, the approach can determine general geometrical configurations of pressured and/or density source and/or sliding structures corresponding to prescribed values of anomalous density, pressure and slip. These source bodies are described as aggregation of elemental point sources for pressure, density and slip, and they fit the whole data (keeping some 3D regularity conditions). In this work we show an application of the methodology to model the real-time evolution of the volcanic source for 2008 eruption of Mount Etna (Italy). To this aim the High-Rate GPS data, coming from the Continuous GPS network, are processed in real-time to obtain sub-daily solutions for tracking the fast dynamics of the magma migration. In our test case we reproduced the real-time scenario of the eruption. Though the data of the test were processed after data collection, real-time operation was emulated. From the results, it is possible to extrapolate the dynamic of a deep and a shallow magma source and the dyke intrusion. In particular, results show at 5 am UTC a magma batch likely migrating towards the surface leaving behind a deflating volume at about 2 km bsl and a deep elongated body from 2 km bsl to 10 km bsl which runs along the High Vp Body and likely represents the deep conduit from where the magma rises up. We demonstrate that the proposed methodology is

  10. Source localization and tracking in a dispersive medium using wireless sensor network

    NASA Astrophysics Data System (ADS)

    Hakim, Kamrul; Jayaweera, Sudharman K.

    2013-12-01

    In this paper, we address the issue of collaborative information processing for diffusive source localization and tracking using wireless sensor networks capable of sensing in dispersive medium/environment. We first determine the space-time concentration distribution of the dispersion from physical modeling and mathematical formulations of an underwater oil spill scenario, considering the effect of laminar water velocity as an external force. For static diffusive source localization, we propose two parametric estimation techniques based on maximum-likelihood (ML) and best linear unbiased estimator for the special case of our physical dispersion model. We prove the consistency and asymptotic normality of the obtained ML solution when the number of sensor nodes and samples approach infinity, and derive the Cramér-Rao lower bound on its performance. We also propose a particle filter-based target tracking scheme for moving diffusive source and derive the posterior Cramér-Rao lower bound for the moving source state estimates as a theoretical performance bound. The performance of the proposed schemes are shown through numerical simulations and compared with the derived theoretical bounds.

  11. 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. PMID:27538026

  12. 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.

  13. Quo vadis source tracking? Towards a strategic framework for environmental monitoring of fecal pollution.

    PubMed

    Santo Domingo, Jorge W; Bambic, Dustin G; Edge, Thomas A; Wuertz, Stefan

    2007-08-01

    Advances in microbial source tracking (MST) have largely been driven by the need to comply with water quality standards based on traditional indicator bacteria. Recently, a number of culture-independent, and library-independent methods based on polymerase chain reaction (PCR) have been gaining popularity among source trackers. However, only a limited number of these methods have been successfully used in field applications, primarily due to the fact that many of them are still being developed. In this critical outlook, we examine different viewpoints associated with the practical use of MST to identify critical research gaps, propose a priority-based timeline to address them, and outline emerging technologies that will likely impact the future of source tracking. We propose that it is necessary to consider each of these aspects in order to advance towards a unifying framework in source identification, so that fecal pollution monitoring can be reliably used for comprehensive environmental microbial monitoring, to develop risk assessment models, and to implement and validate adequate management practices.

  14. 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

  15. First investigations into the prevalence of Cryptosporidium and Giardia spp. in Hungarian drinking water.

    PubMed

    Plutzer, J; Takó, M H; Márialigeti, K; Törökné, A; Karanis, P

    2007-12-01

    Safe drinking water is a top priority in preventing disease outbreaks and is of general concern to everyone. This study examines the occurrence of Cryptosporidium and Giardia in Hungarian drinking water supplies for the first time. A total of 76 raw and drinking water samples were examined using the U.S. EPA Method 1623. From these 15 of 34 (48.4%) raw water samples tested positive for Giardia and 7 (26.6%) for Cryptosporidium. Twelve of 45 (26.7%) drinking water samples were positive for Giardia and 6 (13.3%) for Cryptosporidium. Overall, Giardia cysts and/or Cryptosporidium oocysts were detected in 48% of the raw water samples and 35% of the drinking water samples. The highest levels in drinking water were found to be 3 oocysts/100 litres of Cryptosporidium and 63.6 cysts/100 litres for Giardia, enough to cause giardiasis. The highest levels in raw water were 1,030 cysts/100 litres for Giardia and 50 oocysts/100 litres for Cryptosporidium and higher oocyst densities were associated with source water receiving effluents from sewage treatment plants or originating from a forest environment. In addition to this monitoring, riverbank filtrated water and raw water from the River Danube in Budapest were monitored in order to ascertain protozoan removal efficiency of riverbank filtration (RBF). A total of 157 samples, including 87 samples from the River Danube and 70 samples post RBF, were examined. Cryptosporidium and Giardia were detected regularly in the river water but never in riverbank filtered water suggesting the effectiveness of RBF as a purification method. The occurrence of Cryptosporidium oocysts and Giardia cysts in the investigated water supplies may require the water utilities and water authorities in Hungary to apply additional monitoring and treatment and/or watershed controls.

  16. Intensive exploitation of a karst aquifer leads to Cryptosporidium water supply contamination.

    PubMed

    Khaldi, S; Ratajczak, M; Gargala, G; Fournier, M; Berthe, T; Favennec, L; Dupont, J P

    2011-04-01

    Groundwater from karst aquifers is an important source of drinking water worldwide. Outbreaks of cryptosporidiosis linked to surface water and treated public water are regularly reported. Cryptosporidium oocysts are resistant to conventional drinking water disinfectants and are a major concern for the water industry. Here, we examined conditions associated with oocyst transport along a karstic hydrosystem, and the impact of intensive exploitation on Cryptosporidium oocyst contamination of the water supply. We studied a well-characterized karstic hydrosystem composed of a sinkhole, a spring and a wellbore. Thirty-six surface water and groundwater samples were analyzed for suspended particulate matter, turbidity, electrical conductivity, and Cryptosporidium and Giardia (oo)cyst concentrations. (Oo)cysts were identified and counted by means of solid-phase cytometry (ChemScan RDI(®)), a highly sensitive method. Cryptosporidium oocysts were detected in 78% of both surface water and groundwater samples, while Giardia cysts were found in respectively 22% and 8% of surface water and groundwater samples. Mean Cryptosporidium oocyst concentrations were 29, 13 and 4/100 L at the sinkhole, spring and wellbore, respectively. Cryptosporidium oocysts were transported from the sinkhole to the spring and the wellbore, with respective release rates of 45% and 14%, suggesting that oocysts are subject to storage and remobilization in karst conduits. Principal components analysis showed that Cryptosporidium oocyst concentrations depended on variations in hydrological forcing factors. All water samples collected during intensive exploitation contained oocysts. Control of Cryptosporidium oocyst contamination during intensive exploitation is therefore necessary to ensure drinking water quality.

  17. Evidence of Cryptosporidium transmission between cattle and humans in northern New South Wales.

    PubMed

    Ng, Josephine Su Yin; Eastwood, Keith; Walker, Belinda; Durrheim, David N; Massey, Peter D; Porigneaux, Philippe; Kemp, Ross; McKinnon, Bob; Laurie, Kate; Miller, David; Bramley, Elizabeth; Ryan, Una

    2012-04-01

    Cryptosporidium is an enteric parasite of public health significance that causes diarrhoeal illness through faecal oral contamination and via water. Zoonotic transmission is difficult to determine as most species of Cryptosporidium are morphologically identical and can only be differentiated by molecular means. Transmission dynamics of Cryptosporidium in rural populations were investigated through the collection of 196 faecal samples from diarrheic (scouring) calves on 20 farms and 63 faecal samples from humans on 14 of these farms. The overall prevalence of Cryptosporidium in cattle and humans by PCR and sequence analysis of the 18S rRNA was 73.5% (144/196) and 23.8% (15/63), respectively. Three species were identified in cattle; Cryptosporidium parvum, Cryptosporidium bovis and Cryptosporidium ryanae, and from humans, C. parvum and C. bovis. This is only the second report of C. bovis in humans. Subtype analysis at the gp60 locus identified C. parvum subtype IIaA18G3R1 as the most common subtype in calves. Of the seven human C. parvum isolates successfully subtyped, five were IIaA18G3R1, one was IIdA18G2 and one isolate had a mix of IIaA18G3R1 and IIdA19G2. These findings suggest that zoonotic transmission may have occurred but more studies involving extensive sampling of both calves and farm workers are needed for a better understanding of the sources of Cryptosporidium infections in humans from rural areas of Australia. PMID:22333036

  18. Prevalence and Genetic Characterization of Cryptosporidium Species in Dairy Calves in Central Ethiopia

    PubMed Central

    Wegayehu, Teklu; Karim, Robiul; Anberber, Manyazewal; Adamu, Haileeyesus; Erko, Berhanu; Zhang, Longxian; Tilahun, Getachew

    2016-01-01

    The burden of cryptosporidiosis due to Cryptosporidium parvum is well documented in HIV-positive patients in Ethiopia. However, the role of animals in zoonotic transmission of the disease is poorly understood. The aim of this study was to determine the prevalence and genotypes of Cryptosporidium species in dairy calves; to assess the role of cattle in zoonotic transmission in central Ethiopia. A total of 449 fecal samples were collected and screened using modified Ziehl-Neelson staining method and PCR targeting the small-subunit (SSU) rRNA gene. The prevalence of Cryptosporidium was 9.4% (42/449) and 15.8% (71/449) as detected by microscopy and nested PCR, respectively. The prevalence of infection varied significantly across the study areas with the higher prevalence being observed in Chancho 25.4% (30/118). Crossbred calves had significantly higher prevalence of Cryptosporidium than indigenous zebu. Genotyping results revealed the presence of C. andersoni (76.1%), C. bovis (19.7%) and C. ryanae (4.2%). The occurrence of these Cryptosporidium species appeared to be age-related. C. andersoni constituted 92.1% of the Cryptosporidium infection in calves older than 3 months. Sequence analysis also showed the existence of intra-species variation at SSU rRNA gene. Findings of the current study indicate that cattle may not be an important source of zoonotic cryptosporidiosis in central Ethiopia. Further molecular studies are needed to support this observation from other part of the country. PMID:27135243

  19. Assessment of risk of infection due to Cryptosporidium parvum in drinking water.

    PubMed

    Masago, Y; Katayama, H; Hashimoto, A; Hirata, T; Ohgaki, S

    2002-01-01

    The risk of infection of Cryptosporidium via drinking water was assessed using Monte Carlo simulation with the field survey data of the Sagami River watershed. The levels of Cryptosporidium in this river were found to follow the lognormal distribution. From the counted data, the median level of the Miyayama sampling point was estimated to be 5.7 oocysts per 100L. To calculate the annual risk of infection due to Cryptosporidium in drinking water, the Cryptosporidium level of Miyayama sampling point was used as the water source of the waterworks. The 95% percentile of the annual risk of infection was found to be 10(-2.60). If the daily risk was eliminated when the level of Cryptosporidium in treated water exceeded 1 oocyst per 20L, the 95% percentile of the annual risk was reduced by about 1 log. To reduce the 95% value of the annual risk lower than 10(-4), the risk of days with levels of Cryptosporidium in treated water exceeding 1 oocyst per 80L should be eliminated.

  20. Prevalence and Genetic Characterization of Cryptosporidium Species in Dairy Calves in Central Ethiopia.

    PubMed

    Wegayehu, Teklu; Karim, Robiul; Anberber, Manyazewal; Adamu, Haileeyesus; Erko, Berhanu; Zhang, Longxian; Tilahun, Getachew

    2016-01-01

    The burden of cryptosporidiosis due to Cryptosporidium parvum is well documented in HIV-positive patients in Ethiopia. However, the role of animals in zoonotic transmission of the disease is poorly understood. The aim of this study was to determine the prevalence and genotypes of Cryptosporidium species in dairy calves; to assess the role of cattle in zoonotic transmission in central Ethiopia. A total of 449 fecal samples were collected and screened using modified Ziehl-Neelson staining method and PCR targeting the small-subunit (SSU) rRNA gene. The prevalence of Cryptosporidium was 9.4% (42/449) and 15.8% (71/449) as detected by microscopy and nested PCR, respectively. The prevalence of infection varied significantly across the study areas with the higher prevalence being observed in Chancho 25.4% (30/118). Crossbred calves had significantly higher prevalence of Cryptosporidium than indigenous zebu. Genotyping results revealed the presence of C. andersoni (76.1%), C. bovis (19.7%) and C. ryanae (4.2%). The occurrence of these Cryptosporidium species appeared to be age-related. C. andersoni constituted 92.1% of the Cryptosporidium infection in calves older than 3 months. Sequence analysis also showed the existence of intra-species variation at SSU rRNA gene. Findings of the current study indicate that cattle may not be an important source of zoonotic cryptosporidiosis in central Ethiopia. Further molecular studies are needed to support this observation from other part of the country. PMID:27135243

  1. 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...

  2. 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...

  3. 40 CFR 141.711 - Filtered system additional Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... survey or an equivalent source water assessment that after a system completed the monitoring conducted... lead to increased contamination of the source water by Cryptosporidium, the system must take actions specified by the State to address the contamination. These actions may include additional source...

  4. 40 CFR 141.711 - Filtered system additional Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... survey or an equivalent source water assessment that after a system completed the monitoring conducted... lead to increased contamination of the source water by Cryptosporidium, the system must take actions specified by the State to address the contamination. These actions may include additional source...

  5. 40 CFR 141.711 - Filtered system additional Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... survey or an equivalent source water assessment that after a system completed the monitoring conducted... lead to increased contamination of the source water by Cryptosporidium, the system must take actions specified by the State to address the contamination. These actions may include additional source...

  6. 40 CFR 141.711 - Filtered system additional Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... survey or an equivalent source water assessment that after a system completed the monitoring conducted... lead to increased contamination of the source water by Cryptosporidium, the system must take actions specified by the State to address the contamination. These actions may include additional source...

  7. Assessment of equine fecal contamination: the search for alternative bacterial source-tracking targets.

    PubMed

    Simpson, Joyce M; Santo Domingo, Jorge W; Reasoner, Donald J

    2004-01-01

    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 indicated that upstream and downstream water samples were predominantly beta- and gamma-Proteobacteria (35 and 19%, respectively), while the manure library consisted predominantly of Firmicutes (31%) and previously unidentified sequences (60%). Manure-specific eubacterial sequences were not detectable beyond 5 m downstream of the pile, suggesting either poor survival or high dilution rates. In contrast, Bacteroides and Prevotella sp. sequences were detected both in manure and downstream using group-specific primers. Novel sequences from Bacteroides and Prevotella analysis produced an equine-specific phylogenetic cluster as compared to previous data sets obtained for human and bovine samples. While these results suggest that some anaerobic fecal bacteria might be potential identifiers for use in source-tracking applications, a comprehensive examination of environmental sequences within these species should be performed before methods targeting these bacterial groups are applied to watersheds for development of microbial source-tracking protocols.

  8. 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. PMID:25600418

  9. 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

  10. Complex tensor based blind source separation of EEG for tracking P300 subcomponents.

    PubMed

    Kouchaki, Samaneh; Enshaeifar, Shirin; Cheong Took, Clive; Sanei, Saeid

    2015-08-01

    Complex tensor factorisation of correlated brain sources is addressed in this paper. The electrical brain responses due to motory, sensory, or cognitive stimuli, i.e. event related potentials (ERPs), particularly P300, have been used for cognitive information processing. P300 has two subcomponents, P3a and P3b which are correlated and therefore, the traditional blind source separation approaches cannot solve the problem. In this work, a complex-valued tensor factorisation of electroencephalography (EEG) signals is introduced with the aim of separating P300 subcomponents. The proposed method uses complex-valued statistics to exploit the data correlation. In this way, the variations of P3a and p3b can be tracked for the assessment of the brain state. The results of this work will be compared with those of spatial principal component analysis (SPCA) method. PMID:26737903

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. Cryptosporidium and Giardia detection in water bodies of Galicia, Spain.

    PubMed

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

    2010-12-01

    animal health. These results demonstrate the wide distribution of Cryptosporidium and Giardia in the environment, the ineffectiveness of treatments in DWTPs and WWTPs in reducing/inactivating both protozoa and the need to monitor the presence, viability and infectivity of Cryptosporidium and Giardia in water bodies. In conclusion, the findings suggest the need for better monitoring of water quality and identification of sources of contamination.

  16. 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.

  17. 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.

  18. 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. PMID:22035571

  19. 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...

  20. 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

  1. 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

  2. Modeling increased riverine nitrogen export: Source tracking and integrated watershed-coast management.

    PubMed

    Yu, Dan; Yan, Weijin; Chen, Nengwang; Peng, Benrong; Hong, Huasheng; Zhuo, Guihua

    2015-12-30

    The global NEWS model was calibrated and then used to quantify the long term trend of dissolved inorganic nitrogen (DIN) export from two tributaries of Jiulong River (SE China). Anthropogenic N inputs contributed 61-92% of river DIN yield which increased from 337 in 1980s to 1662 kg N km(-2) yr(-1) in 2000s for the North River, and from 653 to 3097 kg N km(-2) yr(-1) for the West River. North River and West River contributed 55% and 45% respectively of DIN loading to the estuary. Rapid development and poor management driven by national policies were responsible for increasing riverine N export. Scenario analysis and source tracking suggest that reductions of anthropogenic N inputs of at least 30% in the North River (emphasis on fertilizer and manure) and 50% in the West River (emphasis on fertilizer) could significantly improve water quality and mitigate eutrophication in both river and coastal waters. PMID:26517942

  3. Computer analysis of nuclear track emulsion exposed to thermal neutrons and Cf source

    NASA Astrophysics Data System (ADS)

    Mamatkulov, K. Z.; Ambřozová, I.; Artemenkov, D. A.; Bradnova, V.; Kamanin, D. V.; Kattabekov, R. R.; Majling, L.; Marey, A.; Ploc, O.; Rusakova, V. V.; Stanoeva, R.; Turek, K.; Zaitsev, A. A.; Zarubin, P. I.; Zarubina, I. G.

    2016-02-01

    Application of the nuclear track emulsion technique (NTE) in radioactivity and nuclear fission studies is discussed. It is suggested to use a HSP-1000 automated microscope for searching for a collinear cluster tri-partition of heavy nuclei implanted in NTE. Calibrations of α-particles and ion ranges in a novel NTE are carried out. Surface exposures of NTE samples to a Cf-252 source started. Planar events containing fragments and long-range α-particles as well as fragment triples only are studied. Splittings induced by thermal neutrons are studied in boron-enriched emulsion. Use of the image recognition program ”ImageJ” for obtaining characteristics of individual events and for events from the large scan area is presented.

  4. 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

  5. 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.

  6. 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, B.

    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.

  7. 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.

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

    PubMed

    Stewart, Jill R; Boehm, Alexandria B; Dubinsky, Eric A; Fong, Theng-Theng; Goodwin, Kelly D; Griffith, John F; Noble, Rachel T; Shanks, Orin C; Vijayavel, Kannappan; Weisberg, Stephen B

    2013-11-15

    Microbial source tracking (MST) methods were evaluated in the Source Identification Protocol Project (SIPP), in which 27 laboratories compared methods to identify host sources of fecal pollution from blinded water samples containing either one or two different fecal types collected from California. This paper details lessons learned from the SIPP study and makes recommendations to further advance the field of MST. Overall, results from the SIPP study demonstrated that methods are available that can correctly identify whether particular host sources including humans, cows and birds have contributed to contamination in a body of water. However, differences between laboratory protocols and data processing affected results and complicated interpretation of MST method performance in some cases. This was an issue particularly for samples that tested positive (non-zero Ct values) but below the limits of quantification or detection of a PCR assay. Although false positives were observed, such samples in the SIPP study often contained the fecal pollution source that was being targeted, i.e., the samples were true positives. Given these results, and the fact that MST often requires detection of targets present in low concentrations, we propose that such samples be reported and identified in a unique category to facilitate data analysis and method comparisons. Important data can be lost when such samples are simply reported as positive or negative. Actionable thresholds were not derived in the SIPP study due to limitations that included geographic scope, age of samples, and difficulties interpreting low concentrations of target in environmental samples. Nevertheless, the results of the study support the use of MST for water management, especially to prioritize impaired waters in need of remediation. Future integration of MST data into quantitative microbial risk assessments and other models could allow managers to more efficiently protect public health based on site conditions

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

    PubMed

    Stewart, Jill R; Boehm, Alexandria B; Dubinsky, Eric A; Fong, Theng-Theng; Goodwin, Kelly D; Griffith, John F; Noble, Rachel T; Shanks, Orin C; Vijayavel, Kannappan; Weisberg, Stephen B

    2013-11-15

    Microbial source tracking (MST) methods were evaluated in the Source Identification Protocol Project (SIPP), in which 27 laboratories compared methods to identify host sources of fecal pollution from blinded water samples containing either one or two different fecal types collected from California. This paper details lessons learned from the SIPP study and makes recommendations to further advance the field of MST. Overall, results from the SIPP study demonstrated that methods are available that can correctly identify whether particular host sources including humans, cows and birds have contributed to contamination in a body of water. However, differences between laboratory protocols and data processing affected results and complicated interpretation of MST method performance in some cases. This was an issue particularly for samples that tested positive (non-zero Ct values) but below the limits of quantification or detection of a PCR assay. Although false positives were observed, such samples in the SIPP study often contained the fecal pollution source that was being targeted, i.e., the samples were true positives. Given these results, and the fact that MST often requires detection of targets present in low concentrations, we propose that such samples be reported and identified in a unique category to facilitate data analysis and method comparisons. Important data can be lost when such samples are simply reported as positive or negative. Actionable thresholds were not derived in the SIPP study due to limitations that included geographic scope, age of samples, and difficulties interpreting low concentrations of target in environmental samples. Nevertheless, the results of the study support the use of MST for water management, especially to prioritize impaired waters in need of remediation. Future integration of MST data into quantitative microbial risk assessments and other models could allow managers to more efficiently protect public health based on site conditions.

  10. Use of Bacteroidales Microbial Source Tracking To Monitor Fecal Contamination in Fresh Produce Production

    PubMed Central

    Ravaliya, Kruti; 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. PMID:24212583

  11. 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. PMID:24212583

  12. 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

  13. 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.

  14. Molecular epidemiology of Cryptosporidium species in livestock in Ireland.

    PubMed

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

    2016-01-30

    Cryptosporidium is a protozoan that can cause gastro-intestinal illness with diarrhoea in a wide range of hosts. In fact some species of Cryptosporidium can infect the broad range of hosts. The current paper is focused to investigate monthly prevalence and diversity of Cryptosporidium spp. during the spring and early summer (March-June) in 2009 and 2010 in farms with no history of cryptosporidiosis. Animal samples were analyzed to elucidate the prevalence of Cryptosporidium in two regions, West and the East catchments in Ireland. Our investigation demonstrates the prevalence ranges from 14% to 26% an early summer peak (June) was observed. Based on the findings of this study Cryptosporidium ryanae (in cattle, horses), and Cryptosporidium bovis/xiaoi followed by Cryptosporidium parvum (in sheep) were found to be the predominant species in asymptomatic cases. The circulation of other Cryptosporidium species such as C. parvum, C. bovis, C. ubiquitum, C. andersoni and Cryptosporidium horse and pig genotypes in livestock was investigated.

  15. Bacterial source tracking guides management of boat head waste in a coastal resort area.

    PubMed

    Mallin, Michael A; Haltom, Mary I; Song, Bongkeun; Tavares, Mary E; Dellies, Stephen P

    2010-12-01

    Fecal contamination of water bodies causes a public health problem and economic loss. To control such contamination management actions need to be guided by sound science. From 2007-2009 a study was undertaken to determine the sources of fecal bacteria contamination to the marine waters adjoining the Town of Wrightsville Beach, North Carolina, USA. The research effort included sampling for fecal coliform and Enterococcus bacteria, sampling for optical brighteners, dye studies, and use of molecular bacterial source tracking techniques including polymerase chain reaction (PCR) and terminal restriction fragment polymorphism (T-RFLP) fingerprinting of the Bacteroides-Prevotella group. Of the 96 samples collected from nine locations during the study, the water contact standard for Enterococcus was exceeded on 13 occasions. The T-RFLP fingerprint analyses demonstrated that the most widespread source of fecal contamination was human, occurring in 38% of the samples, with secondary ruminant and avian sources also detected. Optical brightener concentrations were low, reflecting a lack of sewage line leakage or spills. A lack of sewer leaks and lack of septic systems in the town pointed toward discharge from boat heads into the marine waters as the major cause of fecal contamination; this was supported by dye studies. Based on these data, the Town initiated action to have the U.S. Environmental Protection Agency declare the coastal waters (out to 3 nautical miles), the nearby Atlantic Intracoastal Waterway and its tributaries a no-discharge zone (NDZ) to alleviate the human fecal pollution. The Town garnered supporting resolutions from other local communities who jointly petitioned the North Carolina Department of Environmental and Natural Resources. This State regulatory agency supported the local government resolutions and sent an application for an NDZ to the EPA in April 2009. The EPA concurred, and in February 2010 the coastal waters of New Hanover County, NC, became the

  16. 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.

  17. 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.

  18. Tracking Paths of Ocean Source Ambient Seismic Noise into, and through, the 3D Earth

    NASA Astrophysics Data System (ADS)

    Reading, A. M.; Gal, M.; Morse, P. E.; Koper, K. D.; Hemer, M. A.; Rawlinson, N.; Salmon, M.; De Kool, M.; Kennett, B. L. N.

    2014-12-01

    Array measurements of seismic noise (microseisms) are emerging as independent observables that inform our knowledge of ocean storms. Using an improved implementation of IAS Capon analysis, we can infer the location and amplitude of multiple sources of seismic noise over multiple decades. For the Southern Ocean, we can use seismic records to assist in identifying shifting patterns of ocean storms. Thus we can investigate topics such as the disparity between wave height trends identified using calibrated satellite records, which appear to be in increasing over multiple decades, and wave heights measured directly using a wave-rider buoy, which does not show a significant change over the same time frame. The passage of wave energy from the water column to the solid Earth, and through the 3D Earth to the seismic array must be tracked effectively. In this contribution, we focus on understanding the passage of seismic noise through the 3D Earth. In particular, we investigate path deviations from 1D Earth models for body waves sources from a variety of locations in the Southern Ocean recorded at Australian seismic arrays. We also investigate path deviations of surface waves travelling across the Australian continent, using the AusREM Earth model. We also appraise other factors affecting the interpretation of slowness, backazimuth and amplitude from seismic array records. These include the effect of the bathymetry-related transfer function controlling energy entering the solid Earth from the water column and the impact of local geology at the site of the seismic array. For a season of storms in the southern hemisphere winter, we simulate the path of energy from a representative range of locations to Australia seismic arrays. We employ a wavefront tracking technique, fast marching, that can support heterogeneous structure and the consideration of multiple arrivals. We find that storms in some locations are subject to a much larger deviation from the expected path of energy

  19. 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.

  20. BacWGSTdb, a database for genotyping and source tracking bacterial pathogens

    PubMed Central

    Ruan, Zhi; Feng, Ye

    2016-01-01

    Whole genome sequencing has become one of the routine methods in molecular epidemiological practice. In this study, we present BacWGSTdb (http://bacdb.org/BacWGSTdb), a bacterial whole genome sequence typing database which is designed for clinicians, clinical microbiologists and hospital epidemiologists. This database borrows the population structure from the current multi-locus sequence typing (MLST) scheme and adopts a hierarchical data structure: species, clonal complex and isolates. When users upload the pre-assembled genome sequences to BacWGSTdb, it offers the functionality of bacterial genotyping at both traditional MLST and whole-genome levels. More importantly, users are told which isolates in the public database are phylogenetically close to the query isolate, along with their clinical information such as host, isolation source, disease, collection time and geographical location. In this way, BacWGSTdb offers a rapid and convenient platform for worldwide users to address a variety of clinical microbiological issues such as source tracking bacterial pathogens. PMID:26433226

  1. 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

  2. Using SWAT, Bacteroidales microbial source tracking markers, and fecal indicator bacteria to predict waterborne pathogen occurrence in an agricultural watershed.

    PubMed

    Frey, Steven K; Topp, Edward; Edge, Thomas; Fall, Claudia; Gannon, Victor; Jokinen, Cassandra; Marti, Romain; Neumann, Norman; Ruecker, Norma; Wilkes, Graham; Lapen, David R

    2013-10-15

    Developing the capability to predict pathogens in surface water is important for reducing the risk that such organisms pose to human health. In this study, three primary data source scenarios (measured stream flow and water quality, modelled stream flow and water quality, and host-associated Bacteroidales) are investigated within a Classification and Regression Tree Analysis (CART) framework for classifying pathogen (Escherichia coli 0157:H7, Salmonella, Campylobacter, Cryptosporidium, and Giardia) presence and absence (P/A) for a 178 km(2) agricultural watershed. To provide modelled data, a Soil Water Assessment Tool (SWAT) model was developed to predict stream flow, total suspended solids (TSS), total N and total P, and fecal indicator bacteria loads; however, the model was only successful for flow and total N and total P simulations, and did not accurately simulate TSS and indicator bacteria transport. Also, the SWAT model was not sensitive to an observed reduction in the cattle population within the watershed that may have resulted in significant reduction in E. coli concentrations and Salmonella detections. Results show that when combined with air temperature and precipitation, SWAT modelled stream flow and total P concentrations were useful for classifying pathogen P/A using CART methodology. From a suite of host-associated Bacteroidales markers used as independent variables in CART analysis, the ruminant marker was found to be the best initial classifier of pathogen P/A. Of the measured sources of independent variables, air temperature, precipitation, stream flow, and total P were found to be the most important variables for classifying pathogen P/A. Results indicate a close relationship between cattle pollution and pathogen occurrence in this watershed, and an especially strong link between the cattle population and Salmonella detections. PMID:24079968

  3. 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.

  4. 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...

  5. 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...

  6. 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...

  7. Cryptosporidium parvum Infection Involving Novel Genotypes in Wildlife from Lower New York State

    PubMed Central

    Perz, Joseph F.; Le Blancq, Sylvie M.

    2001-01-01

    Cryptosporidium, an enteric parasite of humans and a wide range of other mammals, presents numerous challenges to the supply of safe drinking water. We performed a wildlife survey, focusing on white-tailed deer and small mammals, to assess whether they may serve as environmental sources of Cryptosporidium. A PCR-based approach that permitted genetic characterization via sequence analysis was applied to wildlife fecal samples (n = 111) collected from September 1996 to July 1998 from three areas in lower New York State. Southern analysis revealed 22 fecal samples containing Cryptosporidium small-subunit (SSU) ribosomal DNA; these included 10 of 91 white-tailed deer (Odocoileus virginianus) samples, 3 of 5 chipmunk (Tamias striatus) samples, 1 of 2 white-footed mouse (Peromyscus leucopus) samples, 1 of 2 striped skunk (Mephitis mephitis) samples, 1 of 5 racoon (Procyon lotor) samples, and 6 of 6 muskrat (Ondatra zibethicus) samples. All of the 15 SSU PCR products sequenced were characterized as Cryptosporidium parvum; two were identical to genotype 2 (bovine), whereas the remainder belonged to two novel SSU sequence groups, designated genotypes 3 and 4. Genotype 3 comprised four deer-derived sequences, whereas genotype 4 included nine sequences from deer, mouse, chipmunk, and muskrat samples. PCR analysis was performed on the SSU-positive fecal samples for three other Cryptosporidium loci (dihydrofolate reductase, polythreonine-rich protein, and beta-tubulin), and 8 of 10 cloned PCR products were consistent with C. parvum genotype 2. These data provide evidence that there is sylvatic transmission of C. parvum involving deer and other small mammals. This study affirmed the importance of wildlife as potential sources of Cryptosporidium in the catchments of public water supplies. PMID:11229905

  8. 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...

  9. 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...

  10. 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...

  11. Cryptosporidium parvum pig genotype II diagnosed in pigs from the state of Rio de Janeiro, Brazil

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pigs may represent a source of Cryptosporidium sp. infection to humans. The objective of this study was to identify the species present in pigs from the State of Rio de Janeiro, Brazil, and verify what risks pigs represent in transmission of human cryptosporidiosis, since there is no such informati...

  12. Effect of Tillage and Rainfall on Transport of Manure-Applied Cryptosporidium Parvum Oocysts through Soil

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Most waterborne outbreaks of cryptosporidiosis have been attributed to agricultural sources due to the high prevalence of Cryptosporidium oocysts in animal wastes and the practice of spreading manure on farmlands. No-till is an effective soil conservation practice, but it often results in soil havin...

  13. 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...

  14. 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. PMID:26298247

  15. 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.

  16. 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.

  17. 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. PMID:23260177

  18. Source tracking of an anthrax outbreak in northeastern China using complete genome analysis and MLVA genotyping.

    PubMed

    Li, S; An, X; Huang, Y; Pei, G; Cao, D; Mi, Z; Gu, Z; Zhao, X; Li, J; Gu, G; Tong, Y

    2015-01-01

    Anthrax is caused by Bacillus anthracis, an etiological agent behind zoonotic diseases worldwide. B. anthracis is also one of the most dangerous bioterrorism agents. An anthrax outbreak took place in Liaoning Province in northeastern China in August 2012. It resulted in seven human infections and dozens of dead cows. One B. anthracis strain, named Han, was isolated from a dead cow. This strain showed minor pathogenicity in mice and was suspected to be derived from the locally administered vaccine strain, Vac. In order to determine if the Han isolate was derived from the vaccine strain Vac and to track the source of the anthrax outbreak and, so, exclude the possibility of terrorism attack, a complete genome sequencing of these two B. anthracis strains was conducted. With the genome sequencing data, canonical single nucleotide polymorphism (SNP) analysis and whole-genome SNP screening were performed. The results indicate that the Han strain was markedly different from the Vac strain. Further analysis by multiple-locus variable-number tandem repeat analysis (MLVA) showed that Han clustered with previously reported Chinese strains. The result of MLVA15 confirmed that the Han strain is a naturally occurring isolate instead of an engineered agent deliberately distributed by terrorists or other parties. In conclusion, our method used in this study not only facilitates epidemiological studies but also made it easier to distinguish naturally occurring outbreaks from intentionally released pathogens.

  19. 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...

  20. Tracking influential haze source areas in North China using an adjoint model, GRAPES-CUACE

    NASA Astrophysics Data System (ADS)

    An, X. Q.; Zhai, S. X.; Jin, M.; Gong, S. L.; Wang, Y.

    2015-08-01

    Based upon the adjoint theory, the adjoint of the aerosol module in the atmospheric chemical modeling system GRAPES-CUACE (Global/Regional Assimilation and PrEdiction System coupled with the CMA Unified Atmospheric Chemistry Environment) was developed and tested for its correctness. Through statistic comparison, BC (black carbon aerosol) concentrations simulated by GRAPES-CUACE were generally consistent with observations from Nanjiao (one urban observation station) and Shangdianzi (one rural observation station) stations. To track the most influential emission-sources regions and the most influential time intervals for the high BC concentration during the simulation period, the adjoint model was adopted to simulate the sensitivity of average BC concentration over Beijing at the highest concentration time point (referred to as the Objective Function) with respect to BC emission amount over Beijing-Tianjin-Hebei region. Four types of regions were selected based on administrative division and sensitivity coefficient distribution. The adjoint model was used to quantify the effects of emission-sources reduction in different time intervals over different regions by one independent simulation. Effects of different emission reduction strategies based on adjoint sensitivity information show that the more influential regions (regions with relatively larger sensitivity coefficients) do not necessarily correspond to the administrative regions, and the influence effectiveness of sensitivity-oriented regions was greater than the administrative divisions. The influence of emissions on the objective function decreases sharply approximately for the pollutants emitted 17-18 h ago in this episode. Therefore, controlling critical emission regions during critical time intervals on the basis of adjoint sensitivity analysis is much more efficient than controlling administrative specified regions during an experiential time period.

  1. [Spontaneous Cryptosporidium infection in weaned rabbits].

    PubMed

    Pavlásek, I; Lávicka, M; Tůmová, E; Skrivan, M

    1996-12-01

    days of age the occurrence of infection was significantly decreased (3.7%). During the experiment seven rabbits (9.7%) died; six of them (8.3%) at the age of 30-40 days. All these naturally infected and dead animals represented cases of monoinfection with C. parvum. The major clinical signs were typical diarrhoea lasting 3-5 days, inappetency, apathia, lethargy, prominent signs of exhaustion followed by dehydration of the organism. Atrophy of villi of the ileum in one of young rabbits was found histologically. Table II presents concrete data on significantly lower body weights (the decrease being 7-61.5%) as compared with rabbits of the same age not infected with cryptosporidia. It was not possible to evaluate objectively the differences in susceptibility to C. parvum infection between the individual genotypes of rabbits. As was found in the course of our further studies (unpublished data) a possible source of infection of young rabbits can be represented by their mothers in which oocysts are excreted sporadically shortly before parturition and during several days after it. Cryptosporidium infection (cryptosporidiosis) in flocks of broiler rabbits is taken as a new protozoal disease in the Czech Republic and C. parvum as one of possible agents in cases of disorders of digestive tract, namely in rabbits after weaning. PMID:9045499

  2. Cryptosporidium: Genomic and Biochemical Features

    PubMed Central

    Rider, Stanley Dean; Zhu, Guan

    2010-01-01

    Recent progress in understanding the unique biochemistry of the two closely related human enteric pathogens Cryptosporidium parvum and C. hominis has been stimulated by the elucidation of the complete genome sequences for both pathogens. Much of the work that has occurred since that time has been focused on understanding the metabolic pathways encoded by the genome in hopes of providing increased understanding of the parasite biology, and in the identification of novel targets for pharmacological interventions. However, despite identifying the genes encoding enzymes that participate in many of the major metabolic pathways, only a hand full of proteins have actually been the subjects of detailed scrutiny. Thus, much of the biochemistry of these parasites remains a true mystery. PMID:19187778

  3. [Human cryptosporidiosis and Cryptosporidium spp. in Haiti].

    PubMed

    Raccurt, Christian P; Brasseur, Philippe; Verdier, Rose I; Li, Xunde; Eyma, Etna; Stockman, Christine Pannier; Agnamey, Patrice; Guyot, Karine; Totet, Anne; Liautaud, Bernard; Nevez, Gilles; Dei-Cas, Eduardo; Pape, Jean W

    2006-06-01

    Contamination by water-born infectious diseases is closely linked to urban slums conditions such as overcrowding and high level of faecal pollution by animal and human excreta. In this environment, cryptosporidiosis is a major cause of acute diarrhoea in children and chronic persistent diarrhoea in AIDS patients, resulting in increased morbidity and mortality in both populations. The aims of this study conducted in Port-au-Prince, Haiti were to: (i) determine the frequency of Cryptosporidium infection in two populations of patients with diarrhoea, children and AIDS patients, and the existence of Cryptosporidium carriage in healthy adults living in close contact with them; (ii) identify by molecular genotyping the Cryptosporidium species involved; and (iii) evaluate the viability of Cryptosporidium oocysts isolated from human stools. From January 2000 to January 2001, 158 of 1529 diarrhoea stool samples collected from 93 patients with diarrhoea, 57 adults followed at Centres GHESKIO and 36 children admitted at the University Hospital in Port-au-Prince contained Cryptosporidium oocysts (10.3%). The majority of adult patients (98%) were HIV-infected whereas the majority of children (81%) tested negative for HIV. Cryptosporidium was documented in only 1/102 healthy persons living in contact with Cryptosporidium infected patients and infection was with the same genotype as that of the contact patient. Among the 69 Cryptosporidium isolates studied for genotyping, three species were identified: C. hominis (59%), C. parvum (38%) and C. felis (3%). The two C. felis cases are the first reported from AIDS patients in the Caribbean. Most of the children regardless of their HIV status were infected with C. hominis (72%), whereas AIDS patients were more likely to be infected by either human or animal genotypes. These data confirm that immunocompromised individuals are susceptible to a wide range of Cryptosporidium spp. Viability of Cryptosporidium oocysts were determined in an

  4. A foodborne outbreak of Cryptosporidium hominis infection.

    PubMed

    Ethelberg, S; Lisby, M; Vestergaard, L S; Enemark, H L; Olsen, K E P; Stensvold, C R; Nielsen, H V; Porsbo, L J; Plesner, A-M; Mølbak, K

    2009-03-01

    Foodborne outbreaks of cryptosporidiosis are uncommon. In Denmark human cases are generally infrequently diagnosed. In 2005 an outbreak of diarrhoea affected company employees near Copenhagen. In all 99 employees were reported ill; 13 were positive for Cryptosporidium hominis infection. Two analytical epidemiological studies were performed; an initial case-control study followed by a cohort study using an electronic questionnaire. Disease was associated with eating from the canteen salad bar on one, possibly two, specific weekdays [relative risk 4.1, 95% confidence interval (CI) 2.1-8.3]. Three separate salad bar ingredients were found to be likely sources: peeled whole carrots served in a bowl of water, grated carrots, and red peppers (in multivariate analysis, whole carrots: OR 2.1, 95% CI 1.1-4.0; grated carrots: OR 2.1, 95% CI 1.2-3.9; peppers: OR 3.3, 95% CI 1.7-6.6). We speculate that a person excreting the parasite may have contaminated the salad buffet.

  5. Inactivation kinetics of Cryptosporidium parvum oocysts in a swine waste lagoon and spray field.

    PubMed

    Jenkins, Michael B; Liotta, Janice L; Bowman, Dwight D

    2013-04-01

    Because of outbreaks of cryptosporidiosis in humans, some Cryptosporidium spp. have become a public health concern. Commercial swine operations can be a source of this protozoan parasite. Although the species distribution of Cryptosporidium is likely dominated by Cryptosporidium suis , a fraction may be comprised of other Cryptosporidium species infectious to humans such as Cryptosporidium parvum . To better understand the survival dynamics of Cryptosporidium spp., oocysts associated with swine operations, 2 experiments were performed to determine die-off rates of C. parvum oocysts in a swine waste lagoon (2009 and 2010) and its spray field (2010 and 2011). Sentinel chambers containing a lagoon effluent suspension of C. parvum oocysts were submerged in the lagoon, and triplicate chambers were removed over time; oocysts were extracted and assayed for viability. For comparative purposes, inactivation rates of Ascaris suum eggs contained in sentinel chambers were also determined. For 2 spray field experiments, air-dried and sieved surface soil was placed in sentinel chambers, hydrated, and inoculated with a lagoon effluent suspension of C. parvum oocysts. Sentinel chambers and control oocysts in PBS contained in microcentrifuge tubes were buried 1.5 cm below the soil surface in 3 blocks. Triplicate chambers and controls were removed over time; oocysts were extracted and assayed for viability. Based on the first order decay equation, days to reach 99% die-off (T(99)) were determined. T(99)-values determined for the 2 lagoon experiments were 13.1 and 20.1 wk, respectively. A T(99)-value for C. parvum in the spray field was significantly longer at 38.0 wk than the control oocysts in PBS at 29.0 wk. The waste lagoon and spray field system of manure management at this large-scale farrowing operation appeared to reduce the load of C. parvum oocysts before they can be hydrologically transported off the operation and reduces their likelihood of contaminating surface waters

  6. 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

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

    PubMed

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

    2016-04-01

    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.

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

    PubMed

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

    2016-04-01

    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

  9. A simple method for extracting DNA from Cryptosporidium oocysts using the anionic surfactant LSS.

    PubMed

    Sekikawa, Takahiro; Kawasaki, Yu; Katayama, Yasuto; Iwahori, Keisuke

    2011-12-15

    Detection of low amounts of Cryptosporidium oocysts in raw water sources is considered an important component in the management, prevention and control of Cryptosporidium in drinking water supplies as Cryptosporidium causes massive waterborne outbreaks worldwide. As Cryptosporidium has a robust oocyst that is extremely resistant to chlorine and other drinking water disinfectants, both the freeze-thaw method and DNA extraction kits have been commonly used for extracting and purifying DNA from the oocyst. However, the DNA extraction procedures are time consuming and costly. Therefore, a simple and low-cost method to extract and purify DNA from the robust oocyst has been required. In this study, we discussed a simple method for detecting Cryptosporidium DNA with the anionic surfactant, n-lauroylsarcosine sodium salt (LSS) using the loop-mediated isothermal amplification (LAMP) to eliminate the need for the freeze-thaw method and the DNA extraction kits. As a result, Bst DNA polymerase was inhibited by 0.1% LSS but not 0.01% LSS and 5% Triton X-100 or Tween 20. Although DNA was extracted from the oocysts by incubating with 0.1% LSS at 90°C for 15 min, Bst DNA polymerase was inhibited by 0.1% LSS. The inhibition by 0.1% LSS was suppressed by adding 5% of the nonionic surfactants, Triton X-100 or Tween 20. The concentration of LSS in a LAMP tube was 0.01% while that in an incubation tube was 0.1%, because LSS in an incubation tube was diluted by a factor of 10 at the DNA amplification process. Therefore, we found that ten oocysts of Cryptosporidium parvum could be detected by incubation with 0.1% LSS, without removing LSS or adding the nonionic surfactants in the LAMP method. PMID:21924387

  10. 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

    2011-01-01

    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. New cryptosporidium genotypes in HIV-infected persons.

    PubMed Central

    Pieniazek, N. J.; Bornay-Llinares, F. J.; Slemenda, S. B.; da Silva, A. J.; Moura, I. N.; Arrowood, M. J.; Ditrich, O.; Addiss, D. G.

    1999-01-01

    Using DNA sequencing and phylogenetic analysis, we identified four distinct Cryptosporidium genotypes in HIV-infected patients: genotype 1 (human), genotype 2 (bovine) Cryptosporidium parvum, a genotype identical to C. felis, and one identical to a Cryptosporidium sp. isolate from a dog. This is the first identification of human infection with the latter two genotypes. PMID:10341184

  12. A Robust Separating and Tracking Method on Two Wideband Sources by Subspace Rotation with One Vector Hydrophone

    NASA Astrophysics Data System (ADS)

    Wang, De-jun; Li, Feng-hua

    2010-09-01

    It has been proved theoretically that two incompletely correlated sources can be identified by linear signal processing methods. However, it is difficult in practice. A new method to separate two wideband sources with one vector sensor is presented in this paper. The method is the combination of subspace rotation and spatial matched filter. Simulations show that this method is insensitive to the initial azimuth error, independent of signal spectrum, and better man wideband focusing subspace methods at low SNR. The sea trial is performed and the experiment results show that the proposed method is effective to separate and track two wideband sources in the underwater environment.

  13. A Proposal for Source Tracking of Fecal Pollution in Recreational Waters by Pulsed-Field Gel Electrophoresis

    PubMed Central

    Furukawa, Takashi; Suzuki, Yoshihiro

    2013-01-01

    This study aimed to identify specific river sources of fecal contamination by applying pulsed-field gel electrophoresis (PFGE) to environmental water samples from a recreational beach in Japan. The genotypes of all Enterococcus faecium and Enterococcus faecalis strains used as indicators of fecal pollution on the recreational beach and rivers were analyzed by PFGE, and the PFGE profiles of the strains were classified at a 0.9 similarity level using dendrogram analysis. PFGE types of E. faecium isolated from Sakai River or urban drainage were classified in the same cluster. Therefore, the probable sources of fecal pollution on the recreational beach were Sakai River and urban drainage. The approaches for microbial source tracking employed in this study used PFGE with Enterococcus species as an indicator can be a potential tool to specify the source(s) of fecal pollution and contribute to improved public health in coastal environments. PMID:24256972

  14. 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.

  15. 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.

  16. 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

  17. 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...

  18. 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...

  19. 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...

  20. 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...

  1. 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...

  2. 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.

  3. 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.

  4. The first reported cases of human cryptosporidiosis caused by Cryptosporidium hominis in Slovak Republic.

    PubMed

    Ondriska, František; Vrabcová, Ivana; Brinďáková, Silvia; Kváč, Martin; Ditrich, Oleg; Boldiš, Vojtech; Bastlová, Marcela

    2013-01-01

    Cryptosporidiosis belongs to the important parasitic infections with zoonotic potential and the occurrence in European countries is rare. The first cases of cryptosporidiosis caused by Cryptosporidium hominis detected in the Slovak republic were described here. Collection of examined humans consisted of five family members. Faecal specimens were examined by formalin sedimentation, by the Sheather's sugar flotation and by immunochromatography and visualised by the Ziehl-Neelsen acid fast stain. A fragment of the Cryptosporidium small subunit ribosomal RNA gene was amplified by nested polymerase chain reaction and species was determined by restriction fragment length polymorphism analysis with the endonucleases SspI and VspI. C. hominis was found in faeces of two immunocompetent siblings (a 7-year-old boy and a 2-year-old girl). The symptoms occurred only in the boy as gastrointestinal disorders lasting 5 days, and manifested by abdominal pain, an elevated body temperature (37.2 °C), mild diarrhoea, accompanied by lassitude, depression and anorexia. Ultrasonic scan revealed enlarged spleen and mezenteric lymph nodes. Microscopic examination of the stool sample revealed numerous Cryptosporidium oocysts. The DNA typing identified C. hominis subtype IbA10G2. Cryptosporidium was also detected in the boy's sister without any complications and symptoms. Their father, mother and grandmother were parasitologically negative. The source of infection remained unknown. Human cases in present study reflect necessity of systematic attention on intestinal parasites diagnostic inclusive of cryptosporidia.

  5. Molecular characterisation of Cryptosporidium (Apicomplexa) in children and cattle in Romania.

    PubMed

    Vieira, Patricia Manuela; Mederle, Narcisa; Lobo, Maria Luisa; Imre, Kalman; Mederle, Ovidiu; Xiao, Lihua; Darabus, Gheorghe; Matos, Olga

    2015-01-01

    To investigate the transmission of species of Cryptosporidium Tyzzer, 1907 in Timis County, Romania, 48 isolates of Cryptosporidium coccidia from 11 children, 29 calves and eight pigs were characterised by molecular analysis of two loci (SSU rRNA and 60-kDa glycoprotein gene). Overall, 22 isolates were amplified and sequence analyses revealed that all isolates were Cryptosporidium parvum Tyzzer, 1912. Two subtype families were identified, IIa and IId. Subtype IIdA22G1 (n = 4) was the single C. parvum subtype found in children. Subtypes found in calves included IIdA27G1 (n = 8), a novel subtype, IIdA25G1 (n = 5), IIdA22G1 (n = 2), IIdA21G1a (n = 1), and IIaA16G1R1 (n = 1). Subtype IIdA26G1 was found in a pig. These results were significantly different from previous Romanian reports, as the five subtypes of family IId identified in this study were never identified previously in this country. Thus, cattle may be a source of Cryptosporidium infections for humans and the transmission dynamics of C. parvum in Romania is more complex than previously believed.

  6. 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.

  7. Environmental ecology of Cryptosporidium and public health implications.

    PubMed

    Rose, J B

    1997-01-01

    Cryptosporidium has become the most important contaminant found in drinking water and is associated with a high risk of waterborne disease particularly for the immunocompromised. There have been 12 documented waterborne outbreaks in North America since 1985; in two of these (Milwaukee and Las Vegas) mortality rates in the immunocompromised ranged from 52% to 68%. The immunofluorescence antibody assay (IFA) using epifluorescence microscopy has been used to examine the occurrence of Cryptosporidium in sewage (1 to 120 oocysts/liter), filtered secondary treated wastewater (0.01 to 0.13 oocysts/liter), surface waters (0.001 to 107 oocysts/liter), groundwater (0.004 to 0.922 oocysts/liter) and treated drinking water (0.001 to 0.72 oocysts/liter). New rules are being developed (Information Collection Rule and Enhanced Surface Water Treatment Rule) to obtain more occurrence data for drinking water systems for use with new risk assessment models. Public health officials should consider a communication program to physicians treating the immunocompromised, nursing homes, develop a plan to evaluate cases of cryptosporidiosis in the community, and contribute to the development of public policies that limit contamination of source waters, improve water treatment, and protect public health.

  8. 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

  9. Common occurrence of Cryptosporidium hominis in horses and donkeys.

    PubMed

    Jian, Fuchun; Liu, Aiqin; Wang, Rongjun; Zhang, Sumei; Qi, Meng; Zhao, Wei; Shi, Yadong; Wang, Jianling; Wei, Jiujian; Zhang, Longxian; Xiao, Lihua

    2016-09-01

    Extensive genetic variation is observed within the genus Cryptosporidium and the distribution of Cryptosporidium species/genotypes in humans and animals appears to vary by geography and host species. To better understand the genetic diversity of Cryptosporidium spp. in horses and donkeys, we characterized five horse-derived and 82 donkey-derived Cryptosporidium isolates from five provinces or autonomous regions (Sichuan, Gansu, Henan, Inner Mongolia and Shandong) in China at the species/genotype and subtype levels. Three Cryptosporidium species/genotypes were identified based on the analysis of the SSU rRNA gene, including Cryptosporidium parvum (n=22), the Cryptosporidium horse genotype (n=4), and Cryptosporidium hominis (n=61). The identification of C. hominis was confirmed by sequence analysis of the HSP70 and actin genes. Subtyping using sequence analysis of the 60kDa glycoprotein gene identified 21 C. parvum isolates as subtype IIdA19G1, the four horse genotype isolates as subtypes VIaA15G4 (n=2) and VIaA11G3 (n=2), and the 61 C. hominis isolates as IkA16G1 (n=59) and IkA16 (n=2). The common finding of C. hominis reaffirms the heterogeneity of Cryptosporidium spp. in horses and donkeys and is possibly a reflection of endemic transmission of C. hominis in these animals. Data of the study suggest that horses and donkeys as companion animals may potentially transmit Cryptosporidium infections to humans.

  10. Common occurrence of Cryptosporidium hominis in horses and donkeys.

    PubMed

    Jian, Fuchun; Liu, Aiqin; Wang, Rongjun; Zhang, Sumei; Qi, Meng; Zhao, Wei; Shi, Yadong; Wang, Jianling; Wei, Jiujian; Zhang, Longxian; Xiao, Lihua

    2016-09-01

    Extensive genetic variation is observed within the genus Cryptosporidium and the distribution of Cryptosporidium species/genotypes in humans and animals appears to vary by geography and host species. To better understand the genetic diversity of Cryptosporidium spp. in horses and donkeys, we characterized five horse-derived and 82 donkey-derived Cryptosporidium isolates from five provinces or autonomous regions (Sichuan, Gansu, Henan, Inner Mongolia and Shandong) in China at the species/genotype and subtype levels. Three Cryptosporidium species/genotypes were identified based on the analysis of the SSU rRNA gene, including Cryptosporidium parvum (n=22), the Cryptosporidium horse genotype (n=4), and Cryptosporidium hominis (n=61). The identification of C. hominis was confirmed by sequence analysis of the HSP70 and actin genes. Subtyping using sequence analysis of the 60kDa glycoprotein gene identified 21 C. parvum isolates as subtype IIdA19G1, the four horse genotype isolates as subtypes VIaA15G4 (n=2) and VIaA11G3 (n=2), and the 61 C. hominis isolates as IkA16G1 (n=59) and IkA16 (n=2). The common finding of C. hominis reaffirms the heterogeneity of Cryptosporidium spp. in horses and donkeys and is possibly a reflection of endemic transmission of C. hominis in these animals. Data of the study suggest that horses and donkeys as companion animals may potentially transmit Cryptosporidium infections to humans. PMID:27264727

  11. Development of a sensitive method to extract and detect low numbers of Cryptosporidium oocysts from adult cattle faecal samples.

    PubMed

    Wells, B; Thomson, S; Ensor, H; Innes, E A; Katzer, F

    2016-08-30

    Cryptosporidium transmission studies to date have concluded that adult cattle are not a significant source of oocysts contributing to clinical cryptosporidiosis in calves on farm. However current methods of sample processing have been optimised for calf faecal samples and may be less sensitive when used on adult samples due to lower numbers of oocysts and larger size of samples. A modified and novel method of oocyst extraction and concentration was developed and applied in an experiment involving spiking adult cattle faecal samples with known concentrations of Cryptosporidium oocysts. The results showed an increased sensitivity of detection from 100oocysts/g of faecal sample using conventional protocols to 5oocysts/g using the newly developed method. As it is important to be able to accurately assess the contribution of adult ruminants to the transmission of Cryptosporidium, both on farm and in the environment, the development of the techniques described here is likely to make an important contribution to Cryptosporidium transmission studies in future and in subsequent control strategies aimed at the reduction of Cryptosporidium infection in calves on farm. PMID:27523933

  12. Development of a sensitive method to extract and detect low numbers of Cryptosporidium oocysts from adult cattle faecal samples.

    PubMed

    Wells, B; Thomson, S; Ensor, H; Innes, E A; Katzer, F

    2016-08-30

    Cryptosporidium transmission studies to date have concluded that adult cattle are not a significant source of oocysts contributing to clinical cryptosporidiosis in calves on farm. However current methods of sample processing have been optimised for calf faecal samples and may be less sensitive when used on adult samples due to lower numbers of oocysts and larger size of samples. A modified and novel method of oocyst extraction and concentration was developed and applied in an experiment involving spiking adult cattle faecal samples with known concentrations of Cryptosporidium oocysts. The results showed an increased sensitivity of detection from 100oocysts/g of faecal sample using conventional protocols to 5oocysts/g using the newly developed method. As it is important to be able to accurately assess the contribution of adult ruminants to the transmission of Cryptosporidium, both on farm and in the environment, the development of the techniques described here is likely to make an important contribution to Cryptosporidium transmission studies in future and in subsequent control strategies aimed at the reduction of Cryptosporidium infection in calves on farm.

  13. Prevalence and molecular typing of Cryptosporidium in dairy cattle in England and Wales and examination of potential on-farm transmission routes.

    PubMed

    Smith, R P; Clifton-Hadley, F A; Cheney, T; Giles, M

    2014-08-29

    An average of 70 samples were collected from 80 dairy farms in England and Wales, from cattle, co-grazed sheep, wildlife and farm wastes, to investigate prevalence, potential sources and transmission routes of Cryptosporidium. At least one positive sample was detected on 74 of the farms (92.5%) by IFAT microscopy. The prevalence in cattle was 10.2% (95% CI 9.4-11.1%), with greater prevalences detected in calf samples, especially from those under 1 month (45.1%). Young calves were also more likely to be shedding Cryptosporidium parvum and larger concentrations of oocysts, whereas older calves and adult cattle were more likely to be shedding Cryptosporidium bovis and Cryptosporidium andersoni, respectively. The C. parvum subtypes detected were predominantly from types commonly identified in UK cattle (67% were either IIaA15G2R1 or IIaA17G1R1). A novel subtype, IIaA17G1R2, was identified from one cattle sample. The prevalence in co-grazed sheep was low (4%). Birds and rodents may represent significant reservoirs of Cryptosporidium due to high prevalence, large oocyst concentrations, and the detection of a C. parvum subtype known to be present in human populations, identified in samples from these wildlife. Cryptosporidium were detected in dirty water and manure, and also from pasture samples where slurry had been spread. On 64% of the farms, identical Cryptosporidium species were detected (mainly C. parvum or C. bovis) from different cattle groups on the farms, although no direct or indirect contact between the groups were recorded, apart from sharing staff. The same Cryptosporidium species were found in cattle, farm wastes and bird samples on the same farms, but rarely, or not at all, present in sheep or rodent samples. The matching of species/subtypes was also related to the proximity of the different sample sources which may indicate a potential transmission route.

  14. 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

  15. 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...

  16. Infectivity of the cervine genotype of Cryptosporidium

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryptosporidiosis is a diarrheal disease of humans and animals caused by parasites in the genus Cryptosporidium, a genus comprising 19 valid species and 40 genotypes. Most human infections are caused by C. hominis and C. parvum. To a lesser extent infections with C. meleagridis, C. felis, C. canis, ...

  17. 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...

  18. 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

  19. Evaluation of chemical indicators for tracking and apportionment of phosphorus sources to Table Rock Lake in Southwest Missouri, USA.

    PubMed

    Yuan, Zhiwen; Ramaswami, Bala; Casaletto, David; Falke, Stefan; Angenent, Largus T; Giammar, Daniel E

    2007-04-01

    This work evaluated the suitability of selected chemical species as indicators for tracking and apportionment of point and non-point phosphorus sources within the Table Rock Lake watershed in Southwest Missouri, USA. The species were evaluated with respect to their uniqueness to specific source types, their ability to be detected in both sources and receiving waters, and the consistency of their concentration ratios to phosphorus. Four sampling events were conducted at 15 sample locations in one year to collect water samples for measuring the concentrations of total and dissolved phosphorus, seven anions, and 19 major and trace elements. Current analytical methods were sensitive enough for quantification of most of the chemical species in both sources and the receiving waterbody. Due to the high seasonal variation of phosphorus concentrations in source samples, no chemical species had consistent concentration ratios to phosphorus across source types. However, several chemicals were found to be unique to specific sources; bromide ion can be used as a unique indicator for the effluent of the largest wastewater treatment plant in the watershed. PMID:17287001

  20. 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.

  1. 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.

  2. 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

  3. 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

  4. Prevalence of Cryptosporidium spp. and Giardia duodenalis in pigs in Lusaka, Zambia.

    PubMed

    Siwila, Joyce; Mwape, Kabemba E

    2012-01-01

    This study was aimed at determining the prevalence of Cryptosporidium spp. and Giardia duodenalis in pigs which were being raised in intensive management systems. Faecal samples were collected from pigs of all age groups from three different piggery units. Samples were collected directly from the rectum for piglets and weaners and from the floor within 2 min - 5 min of excretion for sows and boars. At the time of collection, faecal consistency was noted as being normal, pasty or diarrhoeic. Samples were analysed further using the Merifluor® Cryptosporidium/Giardia immunofluorescence assay. All piggeries had at least one pig infected with either parasite. From a total 217 samples collected, 96 (44.2%; confidence interval [CI] = 37.6% - 50.9%) were positive for Cryptosporidium spp., whilst 26 (12%; CI = 7.6% - 16.3%) had G. duodenalis parasites. Of all the pigs, 6.9% (15/217) harboured both parasites. With regard to Cryptosporidium spp. infection, statistically significant differences were observed amongst the three units (p = 0.001), whereas no significant differences were observed for G. duodenalis infection (p = 0.13). Prevalence was higher in weaners as compared to other pig classes for both parasites, with significant differences being observed for G. duodenalis infection (p = 0.013). There was, however, no difference in infection between male and female pigs for both parasites. Furthermore, most infections were asymptomatic. From the study results it was clear that Cryptosporidium spp. and G. duodenalis infections were prevalent amongst pigs in the piggeries evaluated and, as such, may act as a source of infection for persons who come into contact with them. PMID:23327315

  5. 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.

  6. 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.

  7. Comparative diagnostic techniques for cryptosporidium infection.

    PubMed

    Omoruyi, Beauty E; Nwodo, Uchechukwu U; Udem, Chukwuneke S; Okonkwo, Francis O

    2014-02-24

    Diarrhoea caused by Cryptosporidium is usually mild in immune competent individuals but severe in the young and those with underlying disease leading to compromised immunity. The conventional diagnosis of Cryptosporidium requires observation of the infective oocysts however, their tiny size yields indistinct results, thus limiting the effectiveness of the conventional diagnostic technique, modified Ziehl-Neelsen (ZN) differential staining. Consequent to the abovementioned limitation, ZN staining, sandwich antigen detection enzyme linked immunosorbent assay (sad-ELISA) and a direct polymerase chain reaction (PCR) assay techniques were evaluated for diagnostic efficacy. Stool samples were collected from 180 consenting adult patients attending outpatient and inpatient clinics at Victoria Hospital, Alice, Eastern Cape Province of South Africa. Subjects were stratified as; 35 HIV-positive and diarrhoeagenic, 125 HIV-negative diarrhoeagenic and 20 apparently healthy controls. Cryptosporidium incidence following diagnostic techniques were 13 (37.1%; ZN staining), 26 (74.3%; sad-ELISA) and 23 (65.7%; PCR), respectively, among HIV-positive diarrhoeagenic patients and 34 (27.2%; ZN staining), 96 (76.8%; sad-ELISA) and 89 (71.2%; PCR) among HIV-negative diarrhoeagenic patients. Sensitivity, specificity and predictive values of the diagnostic techniques' efficiency were: sensitivity: 46.2% (HIV-positive) and 32.3% (HIV-negative) against the ZN technique and 96.9% against sad-ELISA and PCR, respectively, for both HIV-positive and -negative patients; specificity was 88.9% (HIV-positive) and 96.6% (HIV-negative) against the ZN technique. Lastly, the predictive values were 92.3% (HIV-positive) and 96.9% (HIV-negative), respectively, following ZN staining. The sad-ELISA technique proved more suitable for the determination of the presence of Cryptosporidium oocysts. The high incidence of Cryptosporidium in HIV-positive subjects as compared to the HIV-negative population accentuates

  8. 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.

  9. 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. PMID:23731858

  10. 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.

  11. 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...

  12. 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...

  13. 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. PMID:20639363

  14. Concentrations, Viability, and Distribution of Cryptosporidium Genotypes in Lagoons of Swine Facilities in the Southern Piedmont and in Coastal Plain Watersheds of Georgia ▿

    PubMed Central

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

    2010-01-01

    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. PMID:20639363

  15. 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.

  16. Removal of Cryptosporidium by wastewater treatment processes: a review.

    PubMed

    Nasser, Abidelfatah M

    2016-02-01

    Cryptosporidium is a protozoan parasite that infects humans and various animal species. The environmental stability and the low infectious dose of Cryptosporidium facilitate its transmission by water and food. Discharge of untreated wastewater may result in waterborne or foodborne Cryptosporidium outbreaks, therefore a suitable treatment may prevent its dissemination. Most studies on the prevalence of Cryptosporidium oocysts in wastewater have reported a concentration range between 10 and 200 oocysts/L and a prevalence of 6 to 100%. Activated sludge has been found to be ineffective for the removal of Cryptosporidium oocysts. Stabilization ponds and constructed wetlands are efficient for the reduction of Cryptosporidium from wastewater, especially when the retention time is longer than 20 days at suitable sunlight and temperature. High rate filtration and chlorine disinfection are inefficient for the reduction of Cryptosporidium from effluents, whereas ultrafiltration and UV irradiation were found to be very efficient for the reduction of Cryptosporidium oocysts. Adequate tertiary treatment may result in high quality effluent with low risk of Cryptosporidium for unrestricted irrigation and other non-potable applications.

  17. Removal of Cryptosporidium by wastewater treatment processes: a review.

    PubMed

    Nasser, Abidelfatah M

    2016-02-01

    Cryptosporidium is a protozoan parasite that infects humans and various animal species. The environmental stability and the low infectious dose of Cryptosporidium facilitate its transmission by water and food. Discharge of untreated wastewater may result in waterborne or foodborne Cryptosporidium outbreaks, therefore a suitable treatment may prevent its dissemination. Most studies on the prevalence of Cryptosporidium oocysts in wastewater have reported a concentration range between 10 and 200 oocysts/L and a prevalence of 6 to 100%. Activated sludge has been found to be ineffective for the removal of Cryptosporidium oocysts. Stabilization ponds and constructed wetlands are efficient for the reduction of Cryptosporidium from wastewater, especially when the retention time is longer than 20 days at suitable sunlight and temperature. High rate filtration and chlorine disinfection are inefficient for the reduction of Cryptosporidium from effluents, whereas ultrafiltration and UV irradiation were found to be very efficient for the reduction of Cryptosporidium oocysts. Adequate tertiary treatment may result in high quality effluent with low risk of Cryptosporidium for unrestricted irrigation and other non-potable applications. PMID:26837825

  18. 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...

  19. 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...

  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. 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

  2. New molecular quantitative PCR assay for detection of host-specific Bifidobacteriaceae suitable for microbial source tracking.

    PubMed

    Gómez-Doñate, Marta; Ballesté, Elisenda; Muniesa, Maite; Blanch, Anicet R

    2012-08-01

    Bifidobacterium spp. belong to the commensal intestinal microbiota of warm-blooded animals. Some strains of Bifidobacterium show host specificity and have thus been proposed as host-specific targets to determine the origin of fecal pollution. Most strains have been used in microbial-source-tracking (MST) studies based on culture-dependent methods. Although some of these approaches have proved very useful, the low prevalence of culturable Bifidobacterium strains in the environment means that molecular culture-independent procedures could provide practical applications for MST. Reported here is a set of common primers and four Bifidobacterium sp. host-associated (human, cattle, pig, and poultry) probes for quantitative-PCR (qPCR) assessment of fecal source tracking. This set was tested using 25 water samples of diverse origin: urban sewage samples, wastewater from four abattoirs (porcine, bovine, and poultry), and water from a river with a low pollution load. The selected sequences showed a high degree of host specificity. There were no cross-reactions between the qPCR assays specific for each origin and samples from different fecal origins. On the basis of the findings, it was concluded that the host-specific qPCRs are sufficiently robust to be applied in environmental MST studies.

  3. 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.

  4. Tracking the primary sources of fecal pollution in a tropical watershed in a one-year study.

    PubMed

    Toledo-Hernandez, Carlos; Ryu, Hodon; Gonzalez-Nieves, Joel; Huertas, Evelyn; Toranzos, Gary A; Santo Domingo, Jorge W

    2013-03-01

    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 (RGA) watershed were collected mostly on a weekly basis (54 sampling events) during 13 months. DNA extracts from water samples were used in PCR assays to determine the occurrence of fecal bacteria (Bacteroidales, Clostridium coccoides, and enterococci) and human-, cattle-, swine-, and chicken-specific fecal sources. Feces from 12 different animals (n = 340) and wastewater treatment samples (n = 16) were analyzed to determine the specificity and distribution of host-specific assays. The human-specific assay (HF183) was found to be highly specific, as it did not cross-react with nontarget samples. The cattle marker (CF128) cross-reacted to some extent with swine, chicken, and turkeys and was present in 64% of the cattle samples tested. The swine assays showed poor host specificity, while the three chicken assays showed poor host distribution. Differences in the detection of host-specific markers were noted per site. While human and cattle assays showed moderate average detection rates throughout the watershed, areas impacted by wastewater treatment plants and cattle exhibited the highest prevalence of these markers. When conditional probability for positive signals was determined for each of the markers, the results indicated higher confidence levels for the human assay and lower levels for all the other assays. Overall, the results from this study suggest that additional assays are needed, particularly to track cattle, chicken, and swine fecal pollution sources in the RGA watershed. The results also suggest that the geographic stability of genetic markers needs to be determined prior to conducting applied source tracking studies in tropical settings.

  5. Tracking the Primary Sources of Fecal Pollution in a Tropical Watershed in a One-Year Study

    PubMed Central

    Toledo-Hernandez, Carlos; Ryu, Hodon; Gonzalez-Nieves, Joel; Huertas, Evelyn; Toranzos, Gary A.

    2013-01-01

    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 (RGA) watershed were collected mostly on a weekly basis (54 sampling events) during 13 months. DNA extracts from water samples were used in PCR assays to determine the occurrence of fecal bacteria (Bacteroidales, Clostridium coccoides, and enterococci) and human-, cattle-, swine-, and chicken-specific fecal sources. Feces from 12 different animals (n = 340) and wastewater treatment samples (n = 16) were analyzed to determine the specificity and distribution of host-specific assays. The human-specific assay (HF183) was found to be highly specific, as it did not cross-react with nontarget samples. The cattle marker (CF128) cross-reacted to some extent with swine, chicken, and turkeys and was present in 64% of the cattle samples tested. The swine assays showed poor host specificity, while the three chicken assays showed poor host distribution. Differences in the detection of host-specific markers were noted per site. While human and cattle assays showed moderate average detection rates throughout the watershed, areas impacted by wastewater treatment plants and cattle exhibited the highest prevalence of these markers. When conditional probability for positive signals was determined for each of the markers, the results indicated higher confidence levels for the human assay and lower levels for all the other assays. Overall, the results from this study suggest that additional assays are needed, particularly to track cattle, chicken, and swine fecal pollution sources in the RGA watershed. The results also suggest that the geographic stability of genetic markers needs to be determined prior to conducting applied source tracking studies in tropical settings. PMID:23291547

  6. 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.

  7. Tracking Signal Subspace Invariance for Blind Separation and Classification of Nonorthogonal Sources in Correlated Noise

    NASA Astrophysics Data System (ADS)

    Oweiss, Karim G.; Anderson, David J.

    2006-12-01

    We investigate a new approach for the problem of source separation in correlated multichannel signal and noise environments. The framework targets the specific case when nonstationary correlated signal sources contaminated by additive correlated noise impinge on an array of sensors. Existing techniques targeting this problem usually assume signal sources to be independent, and the contaminating noise to be spatially and temporally white, thus enabling orthogonal signal and noise subspaces to be separated using conventional eigendecomposition. In our context, we propose a solution to the problem when the sources are nonorthogonal, and the noise is correlated with an unknown temporal and spatial covariance. The approach is based on projecting the observations onto a nested set of multiresolution spaces prior to eigendecomposition. An inherent invariance property of the signal subspace is observed in a subset of the multiresolution spaces that depends on the degree of approximation expressed by the orthogonal basis. This feature, among others revealed by the algorithm, is eventually used to separate the signal sources in the context of "best basis" selection. The technique shows robustness to source nonstationarities as well as anisotropic properties of the unknown signal propagation medium under no constraints on the array design, and with minimal assumptions about the underlying signal and noise processes. We illustrate the high performance of the technique on simulated and experimental multichannel neurophysiological data measurements.

  8. Cryptosporidium Parvum Transport Through Natural Porous Media

    NASA Astrophysics Data System (ADS)

    Araujo, J. B.; Santamaria, J.; Blandford, W. P.; Gerba, C. P.; Brusseau, M. L.

    2005-12-01

    The objective of this study was to quantify the transport of Cryptosporidium parvum through saturated natural porous media. A series of miscible-displacement experiments were conducted, varying the properties of the porous media and electrolyte solution to help elucidate retention mechanisms. Significant removal (~99%) of oocysts was observed for transport in a sandy soil. Similar removals were also observed for experiments conducted with deionized water in place of the 0.01M NaCl electrolyte solution and experiments with a sub sample of the sandy soil that was treated with nitric acid. Effluent recoveries were greater for experiments conducted using coarser porous media. These results indicate straining contributed to the retention of Cryptosporidium parvum in our system.

  9. 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

    2015-02-01

    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.

  10. Detection of Cryptosporidium parvum in soil extracts.

    PubMed

    Walker, Mark; Redelman, Douglas

    2004-03-01

    Epifluorescent microscopy and flow cytometry were used in different combinations with fluorescein isothiocyanate-labeled immunoglobulins M and G3 to estimate the numbers of Cryptosporidium parvum oocysts in soil extracts containing 10 to 10,017 oocysts/ml. No combination had a systematic effect on accuracy or precision. Background debris may have produced overestimates at low oocyst concentrations when flow cytometry was used. PMID:15006810

  11. 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...

  12. 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

  13. Coherent source imaging and dynamic support tracking for inverse scattering using compressive MUSIC

    NASA Astrophysics Data System (ADS)

    Lee, Okkyun; Kim, Jong Min; Yoo, Jaejoon; Jin, Kyunghwan; Ye, Jong Chul

    2011-09-01

    The goal of this paper is to develop novel algorithms for inverse scattering problems such as EEG/MEG, microwave imaging, and/or diffuse optical tomograpahy, and etc. One of the main contributions of this paper is a class of novel non-iterative exact nonlinear inverse scattering theory for coherent source imaging and moving targets. Specifically, the new algorithms guarantee the exact recovery under a very relaxed constraint on the number of source and receivers, under which the conventional methods fail. Such breakthrough was possible thanks to the recent theory of compressive MUSIC and its extension using support correction criterion, where partial support are estimated using the conventional compressed sensing approaches, then the remaining supports are estimated using a novel generalized MUSIC criterion. Numerical results using coherent sources in EEG/MEG and dynamic targets confirm that the new algorithms outperform the conventional ones.

  14. Cryptosporidium parvum: From foal to veterinary students.

    PubMed

    Galuppi, R; Piva, S; Castagnetti, C; Sarli, G; Iacono, E; Fioravanti, M L; Caffara, M

    2016-03-30

    This paper describes the transmission of a zoonotic subtype of Cryptosporidium parvum between two foals hospitalized in an Equine Perinatology Unit (EPU) linked to an outbreak of cryptosporidiosis in veterinary students. Fecal specimens of 36 mares (105 samples) and 28 foals (122 samples) were subjected to Ziehl-Neelsen staining, nested PCR of 18S rDNA. Two foals tested positive for Cryptosporidium; PCR restriction fragment length polymorphism (PCR-RFLP) analysis and subtyping by nested PCR of the 60kDa glycoprotein (gp60) gene revealed C. parvum subtype IIdA23G1. The introduction of Cryptosporidium into the EPU is suspected to be in a foal showing no initial clinical signs that tested positive for C. parvum during an asymptomatic phase. A second foal, hospitalized afterwards for perinatal asphyxia syndrome complicated with failure of passive transfer and sepsis, showed severe watery diarrhea after 4 days of hospitalization and was positive for the same subtype. During this period, six students attending the EPU complained of abdominal pain and diarrhea and were positive for the same subtype of C. parvum. To the authors' knowledge, this is the first description of this subtype in foals and the first report of evidence of zoonotic transmission of cryptosporidiosis from foals to human. PMID:26921039

  15. Additional novel Cryptosporidium genotypes in ornamental fishes.

    PubMed

    Morine, M; Yang, R; Ng, J; Kueh, S; Lymbery, A J; Ryan, U M

    2012-12-21

    Current knowledge on the prevalence and genotypes of Cryptosporidium in fishes is still limited. This study investigated the prevalence of Cryptosporidium species in 171 ornamental fishes, belonging to 33 species, collected from 8 commercial aquariums around Perth, Western Australia. All samples were screened by nested PCR targeting the 18S rRNA locus. A total of 6 positives were identified by PCR at the 18S locus from 4 different species of fishes (red eye tetra, Moenkhausia sanctaefilomenae; gold gourami, Trichogaster trichopterus; neon tetra, Paracheirodon innesi; goldfish, Carassius auratus auratus), giving an overall prevalence of 3.5% (6/171). Four different genotypes were identified, only one of which has been previously reported in fish; piscine genotype 4 in a neon tetra isolate, a rat genotype III-like isolate in a goldfish, a novel genotype in three isolates from red eye (piscine genotype 7) which exhibited a 3.5% genetic distance from piscine genotype 1 and a piscine genotype 6-like from a gold gourami (1% genetic distance). Further biological and genetic characterisation is required to determine the relationship of these genotypes to established species and strains of Cryptosporidium. PMID:22819587

  16. Tracking Nonpoint Source Nitrogen and Carbon in Watersheds of Chesapeake Bay

    NASA Astrophysics Data System (ADS)

    Kaushal, S.; Pennino, M. J.; Duan, S.; Blomquist, J.

    2012-12-01

    Humans have altered nitrogen and carbon cycles in rivers regionally with important impacts on coastal ecosystems. Nonpoint source nitrogen pollution is a leading contributor to coastal eutrophication and hypoxia. Shifts in sources of carbon impact downstream ecosystem metabolism and fate and transport of contaminants in coastal zones. We used a combination of stable isotopes and optical tracers to investigate fate and transport of nitrogen and carbon sources in tributaries of the largest estuary in the U.S., the Chesapeake Bay. We analyzed isotopic composition of water samples from major tributaries including the Potomac River, Susquehanna River, Patuxent River, and Choptank River during routine and storm event sampling over multiple years. A positive correlation between δ15N-NO3- and δ18O-NO3- in the Potomac River above Washington D.C. suggested denitrification or biological uptake in the watershed was removing agriculturally-derived N during summer months. In contrast, the Patuxent River in Maryland showed elevated δ15N-NO3- (5 - 12 per mil) with no relationship to δ18O-NO3- suggesting the importance of wastewater sources. From the perspective of carbon sources, there were distinct isotopic values of the δ13C-POM of particulate organic matter and fluorescence excitation emission matrices (EEMS) for rivers influenced by their dominant watershed land use. EEMS showed that there were increases in the humic and fulvic fractions of dissolved organic matter during spring floods, particularly in the Potomac River. Stable isotopic values of δ13C-POM also showed rapid depletion suggesting terrestrial carbon "pulses" in the Potomac River each spring. The δ15N-POM peaked to 10 - 15 per mil each spring suggested a potential manure source or result of biological processing within the watershed. Overall, there were considerable changes in sources and transformations of nitrogen and carbon that varied across rivers and that contribute to nitrogen and carbon loads

  17. 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...

  18. Clinical and subclinical infections with Cryptosporidium in animals

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryptosporidium spp. are frequent parasites of livestock and companion animals, raising questions about the clinical significance of such infections. Cryptosporidium infections have a wide spectrum of symptoms that can vary from asymptomatic to serious infection to death. This review presents the cu...

  19. A simplified low-cost approach to antibiotic resistance profiling for faecal source tracking.

    PubMed

    Ebdon, J E; Wallis, J L; Taylor, H D

    2004-01-01

    Antibiotic resistance profiling (ARP) is a potentially useful method for distinguishing faecal bacteria according to host source. This phenotypic approach has cost benefits over genotypic methods, but existing protocols are time-consuming and manual data handling is open to human error. A simplified, low-cost approach to the ARP technique was developed that used automated data recording techniques combined with simple statistical analyses to compare isolates of the genus Enterococcus from various faecal sources. An initial battery of 21 antibiotics (at up to four concentrations) was chosen for source discrimination. Images of growth or non-growth in microplate wells were stored as bitmaps and converted to binary data to form a database of known antibiotic resistance profiles. Discriminant function analysis (DFA) showed that the average rate of isolates correctly classified by the database (wastewater vs non-wastewater) was 86%. Once the more discriminating antibiotics and their concentrations had been identified, it was possible to reduce the number of tests from 80 to 18 whilst increasing the number of correctly classified human isolates. ARP could offer a low-cost and rapid means of identifying sources of faecal pollution. As such, the technique may be of particular benefit to developing countries, where water quality may have a significant impact on health and where cost is a major factor when choosing environmental management technology. PMID:15318507

  20. 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.

  1. 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...

  2. 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...

  3. 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...

  4. 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...

  5. Spatial distribution of risk factors for Cryptosporidium spp. transport in an Irish catchment.

    PubMed

    Samadder, S R; Ziegler, P; Murphy, T M; Holden, N M

    2010-08-01

    Cryptosporidium spp. has become a major public health concern in many parts of the globe, including Ireland, as a result of recent reported waterborne outbreaks of Cryptosporidiosis. Continuous monitoring of Cryptosporidium spp. in water supplies is not feasible, so a risk-forecasting approach is required. This study reports a globally applicable approach for evaluating the spatial variation in relative risk of contaminating surface water by Cryptosporidium spp. based on a risk potential index (RPI) as an indicator of the potential pollution of surface water. The RPI is predicted by readily available data on land use, rainfall, soil type, slope, soil moisture deficit, and distance from water course. A small catchment in County Meath, Ireland, was chosen to illustrate the analysis of the approach. Data for the study area were digitized and rectified using surveyed ground control points to capture each of the RPI factors, field boundaries, and land use. The six parameters were classified and assigned a relative risk score out of 5. A Geographic Information Systems overlay analysis then was used to calculate a cumulative relative risk score for each month of the year. The analysis indicated that April and June experienced a relatively low risk of Cryptosporidium spp. transport compared with other months of the year. June had the least risk, because more than 98% of the catchment was estimated to be of low or moderate risk (RPI ranges = 0 to 2). December had the highest risk of Cryptosporidium spp. transport, because approximately 20% of the catchment area had a moderately high to very high risk (RPI ranges = 2 to 5). The study also made an attempt to reduce the risk of contaminating surface water by alternative land-use practice and relocating the field boundaries. The study demonstrated a semi-quantitative and readily implemented method for using spatial risk assessment for planning land management to reduce the risk of surface water contamination by Cryptosporidium

  6. 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.

  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. PMID:23478072

  8. Occurrence of Cryptosporidium sp. in snakes in Japan.

    PubMed

    Kuroki, Toshiro; Izumiyama, Shinji; Yagita, Kenji; Une, Yumi; Hayashidani, Hideki; Kuro-o, Masaki; Mori, Akira; Moriguchi, Hajime; Toriba, Michihisa; Ishibashi, Toru; Endo, Takuro

    2008-09-01

    The aim of this study was to determine the prevalence of Cryptosporidium in snakes in Japan. Fecal samples or intestinal contents of 469 snakes, consisting of five species, were analyzed and Cryptosporidium oocysts were detected only from the Japanese grass snake Rhabdophis tigrinus. The mean prevalence of Cryptosporidium sp. in Japanese grass snakes was approximately 26% in the region studied. Histopathological observations revealed that the organism caused proliferative enteritis in the small intestine. Sequence analysis of a fragment of the small subunit rRNA gene has shown that the partial sequence of Cryptosporidium sp. isolated from the snakes was identical to that of the Cryptosporidium snake genotype W11 from New Guinea viper boa.

  9. Diagnostic and analytical mutation scanning of Cryptosporidium: utility and advantages.

    PubMed

    Jex, Aaron R; Gasser, Robin B

    2009-03-01

    Cryptosporidiosis is predominantly a disease of the alimentary tract of humans and other vertebrates, caused by parasitic protists of the genus Cryptosporidium. This disease, transmitted mainly via the fecal-oral route (in water or food), is of major socioeconomic importance globally. The diagnosis of cryptosporidiosis, including the genetic characterization of the different species, genotypes and subgenotypes (population variants) of Cryptosporidium, is crucial to prevention and control, particularly as there is no cost-effective treatment against this disease. Although traditional phenetic techniques have had major deficiencies for the specific diagnosis of cryptosporidiosis, there has been substantial progress in the establishment of molecular tools. In this article, we review key genetic markers used for the specific identification of Cryptosporidium, diagnosis of cryptosporidiosis and analysis of genetic variation in Cryptosporidium populations. We also discuss the advantages and disadvantages of selected techniques, and emphasize the benefits of utilizing rapid mutation scanning in achieving improved insights into the population genetics and epidemiology of Cryptosporidium.

  10. 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.

  11. Tracking the sources of nitrate in groundwater using coupled nitrogen and boron isotopes: a synthesis.

    PubMed

    Widory, David; Petelet-Giraud, Emmanuelle; Négrel, Philippe; Ladouche, Bernard

    2005-01-15

    Nitrate (NO3) is one of the world's major pollutants of drinking water resources. Although recent European Directives have reduced input from intensive agriculture, NO3 levels in groundwater are approaching the drinking water limit of 50 mg L(-1) almost everywhere. Determining the sources of groundwater contamination is an important first step toward improving its quality by emission control. It is with this aim that we review here the benefit of using a coupled isotopic approach (delta15N and delta11B), in addition to conventional hydrogeological analyses, to trace the origin of NO3 in water. The studied watersheds include both fractured bedrock and alluvial (subsurface and deep) hydrogeological contexts. The joint use of nitrogen and boron isotope systematics in each context deciphers the origin of NO3 in the groundwater and allows a semi-quantification of the contributions of the respective pollution sources (mineral fertilizers, wastewater, and animal manure).

  12. 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.

  13. 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.

  14. Open Source Tracking and Analysis of Adult Drosophila Locomotion in Buridan's Paradigm with and without Visual Targets

    PubMed Central

    Colomb, Julien; Reiter, Lutz; Blaszkiewicz, Jedrzej; Wessnitzer, Jan; Brembs, Bjoern

    2012-01-01

    Background Insects have been among the most widely used model systems for studying the control of locomotion by nervous systems. In Drosophila, we implemented a simple test for locomotion: in Buridan's paradigm, flies walk back and forth between two inaccessible visual targets [1]. Until today, the lack of easily accessible tools for tracking the fly position and analyzing its trajectory has probably contributed to the slow acceptance of Buridan's paradigm. Methodology/Principal Findings We present here a package of open source software designed to track a single animal walking in a homogenous environment (Buritrack) and to analyze its trajectory. The Centroid Trajectory Analysis (CeTrAn) software is coded in the open source statistics project R. It extracts eleven metrics and includes correlation analyses and a Principal Components Analysis (PCA). It was designed to be easily customized to personal requirements. In combination with inexpensive hardware, these tools can readily be used for teaching and research purposes. We demonstrate the capabilities of our package by measuring the locomotor behavior of adult Drosophila melanogaster (whose wings were clipped), either in the presence or in the absence of visual targets, and comparing the latter to different computer-generated data. The analysis of the trajectories confirms that flies are centrophobic and shows that inaccessible visual targets can alter the orientation of the flies without changing their overall patterns of activity. Conclusions/Significance Using computer generated data, the analysis software was tested, and chance values for some metrics (as well as chance value for their correlation) were set. Our results prompt the hypothesis that fixation behavior is observed only if negative phototaxis can overcome the propensity of the flies to avoid the center of the platform. Together with our companion paper, we provide new tools to promote Open Science as well as the collection and analysis of digital

  15. 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.

  16. Effect of hillslope position and manure application rates on the persistence of fecal source tracking indicators in an agricultural soil.

    PubMed

    Piorkowski, Gregory S; Bezanson, Greg S; Jamieson, Rob C; Hansen, Lisbeth Truelstrup; Yost, Chris K

    2014-03-01

    The influence of liquid dairy manure (LDM) application rates (12.5 and 25 kL ha) and soil type on the decay rates of library-independent fecal source tracking markers (host-associated and mitochondrial DNA) and persistent (>58 d) population structure was examined in a field study. The soils compared were an Aquic Haplorthod and a Typic Haplorthod in Nova Scotia, Canada, that differed according to landscape position and soil moisture regime. Soil type and LDM application rate did not influence decay rates (0.045-0.057 d). population structure, in terms of the occurrence of abundance of strain types, varied according to soil type ( = 0.012) but did not vary by LDM application rate ( = 0.121). Decay of ruminant-specific (BacR), bovine-specific (CowM2), and mitochondrial DNA (AcytB) markers was analyzed for 13 d after LDM application. The decay rates of BacR were greater under high-LDM application rates (0.281-0.358 d) versus low-LDM application rates (0.212-0.236 d) but were unaffected by soil type. No decay rates could be calculated for the CowM2 marker because it was undetectable within 6 d after manure application. Decay rates for AcytB were lower for the Aquic Haplorthod (0.088-0.100 d), with higher moisture status compared with the Typic Haplorthod (0.135 d). Further investigation into the decay of fecal source tracking indicators in agricultural field soils is warranted to assess the influence of soil type and agronomic practice on the differential decay of relevant markers and the likelihood of transport in runoff.

  17. 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

  18. Array tracking of the volcanic tremor source at Deception Island, Antarctica

    NASA Astrophysics Data System (ADS)

    Almendros, J.; Ibáñez, J. M.; Alguacil, G.; Del Pezzo, E.; Ortiz, R.

    We have found experimental evidence which shows that the volcanic tremor recorded at Deception Island (South Shetland Islands, Antarctica) is a superposition in time of overlapping hybrid events. We studied data from a small aperture seismic array. Data analysis for tremor and hybrids included: (1) spectral analysis; (2) apparent slowness and back-azimuth determination by using the zero-lag cross-correlation method; and (3) polarization analysis. Both types of events share these common features: (a) two dominant spectral bands at frequencies 1-3 Hz (the most energetic) and 4-8 Hz; (b) several coherent phases with the same back-azimuth to the source and apparent slowness along the whole signal; (c) in the high frequency band, the apparent slowness is very low (around 0.17 s/km), indicating the propagation of body waves; (d) in the low frequency band, the apparent slowness is high (around 1.6 s/km), consistent with the presence of surface waves; and (e) clear P-wave onset followed by a complex pattern of Rayleigh waves. Therefore, both types of events are strongly related because they share the same source region, the same wave-propagation properties, and the same wave composition. Moreover, several arrivals, that resemble a single hybrid event, have been found along the tremor signals. Due to these reasons, we hypothesize that volcanic tremor of Deception Island is a superposition of hybrid type events. The source of both types could be the interaction between thaw water and hot materials in a shallow aquifer.

  19. Tracking the sources of salmonella in ground beef produced from nonfed cattle.

    PubMed

    Koohmaraie, Mohammad; Scanga, John A; De La Zerda, Michael J; Koohmaraie, Bijan; Tapay, Lourdes; Beskhlebnaya, Vika; Mai, Tam; Greeson, Kay; Samadpour, Mansour

    2012-08-01

    The objective of this study was to determine the source(s) of Salmonella contamination in ground beef. One hundred dairy cows were harvested in a U.S. commercial beef processing plant. Samples of hides, carcasses after hide removal and before exposure to antimicrobial intervention, carcasses after all antimicrobial interventions, superficial cervical lymph nodes from the chuck, trim, ground beef, and air were obtained. Ninety-six percent of the hide samples, 47% of the carcasses before intervention, 18% of the lymph nodes, 7.14% of the trim, and 1.67% of the ground beef samples were positive for Salmonella. None of the samples obtained from the carcasses after the full complement of interventions and none of the air samples were positive for Salmonella. All Salmonella-positive samples were subjected to pulsed-field gel electrophoresis, and eight DNA Xba I restriction patterns were identified. The majority of isolates had one of two restriction digest patterns. The strain isolated from ground beef had the same pattern as the strains isolated from hides and from carcasses immediately after hide removal. The Salmonella isolates from trim samples and lymph nodes also had the same restriction digest pattern. These results indicate that hide and lymph nodes are the most likely sources of Salmonella in ground beef. Dressing practices that effectively reduce or eliminate the transfer of bacteria from hide to carcass and elimination of lymph nodes as a component of raw ground beef should be considered as measures to reduce Salmonella contamination of ground beef. Because total elimination of lymph nodes from ground beef is not possible, other approaches should be explored. Easily accessible lymph nodes could be screened for Salmonella very early in the slaughter process. When the results are positive for Salmonella, the corresponding carcasses should be fabricated separately at the end of the production run, and the trim from these carcasses should be subjected to a

  20. 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.

  1. Unusual cryptosporidiosis cases in Swedish patients: extended molecular characterization of Cryptosporidium viatorum and Cryptosporidium chipmunk genotype I.

    PubMed

    Lebbad, Marianne; Beser, Jessica; Insulander, Mona; Karlsson, Lillemor; Mattsson, Jens G; Svenungsson, Bo; Axén, Charlotte

    2013-12-01

    Most human cases of cryptosporidiosis are caused by Cryptosporidium parvum or Cryptosporidium hominis, but the use of molecular diagnostic methods has revealed that several other less common species or genotypes can also be involved. Here, we describe two unusual causes of cryptosporidiosis, one being the recently described species Cryptosporidium viatorum and the other Cryptosporidium chipmunk genotype I. Two Swedish patients who were infected with C. viatorum had travelled to Kenya and Guatemala, respectively, and two others had been infected with Cryptosporidium chipmunk genotype I in Sweden. None of these four patients were immunocompromised, and all four showed classical symptoms of cryptosporidiosis. We performed extensive molecular characterization, including analysis of four loci. The two C. viatorum isolates were found to differ slightly at the 70-kDa heat shock protein locus, which may indicate a local geographical variation in this species that has previously been described exclusively on the Indian subcontinent.

  2. 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.

  3. 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. PMID:26818945

  4. 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

  5. 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

  6. 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.

  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

    2016-03-01

    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

  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. 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

  10. Recent innovation in microbial source tracking using bacterial real-time PCR markers in shellfish.

    PubMed

    Mauffret, A; Mieszkin, S; Morizur, M; Alfiansah, Y; Lozach, S; Gourmelon, M

    2013-03-15

    We assessed the capacity of real-time PCR markers to identify the origin of contamination in shellfish. Oyster, cockles or clams were either contaminated with fecal materials and host-associated markers designed from Bacteroidales or Catellicoccus marimammalium 16S RNA genes were extracted from their intravalvular liquid, digestive tissues or shellfish flesh. Extraction of bacterial DNA from the oyster intravalvular liquid with FastDNA spin kit for soil enabled the selected markers to be quantified in 100% of artificially contaminated samples, and the source of contamination to be identified in 13 out of 38 naturally contaminated batches from European Class B and Class C areas. However, this protocol did not enable the origin of the contamination to be identified in cockle or clam samples. Although results are promising for extracts from intravalvular liquid in oyster, it is unlikely that a single protocol could be the best across all bacterial markers and types of shellfish. PMID:23398745

  11. 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.

  12. Prevalence and molecular epidemiology of Cryptosporidium parvum in dairy calves in Punjab (India).

    PubMed

    Joute, J R; Gill, J P S; Singh, B B

    2016-09-01

    Cryptosporidium parvum is an important zoonotic protozoan parasite that infects the gastrointestinal tract of vertebrate animals and man. The current study was contemplated for molecular detection of Cryptosporidium species prevalent in dairy calves in Punjab, India. A total of 302 faecal samples were screened by modified Ziehl-Neelsen staining technique for the detection of Cryptosporidium oocysts. Molecular characterisation was done using PCR followed by sequence analysis of the representative isolates. An overall prevalence of 26.15 % was obtained with the highest prevalence obtained in 0-30 day old calves in both diarrhoeic and non-diarrhoeic animals. PCR analysis revealed the expected bands at 1,325 and 835 bp from all the isolates for primary and secondary/nested PCR respectively. Ten representative samples were sequenced in both directions. Phylogenetic analysis revealed the presence of C. parvum in all the samples. The high rate of calves infected with C. parvum can act as a great source of zoonotic cryptosporidiosis which indicates a potential risk of zoonotic transmission from animal to human beings in Punjab (India). PMID:27605777

  13. Point prevalence of Cryptosporidium oocyst in calves grazing along River Rima bank in Sokoto, Nigeria.

    PubMed

    Faleke, O O; Yabo, Y A; Olaleye, A O; Dabai, Y U; Ibitoye, E B

    2014-02-01

    The present study was conducted to investigate the point prevalence of Cryptosporidium oocysts infection in calves grazing along the bank of Rima River Sokoto in October 2011. The river bank is a converging zone for domestic animals reared in different quarters of the town and the surrounding settlements. A total number of 2,959 cattle were enumerated out of which 147 (4.97%) were calves. Faecal samples were collected from 100 (68.02%) calves by convenient sampling technique. Formol-Ether sedimentation and modified Ziehl-Neelsen staining techniques were used to identify the Cryptosporidium oocysts in the faecal samples. Faecal consistency was also used to identify diarrhoeic and non-diarrhoeic calves. Cryptosporidium oocysts were identified in 33 (33.0%) of the calves examined. The detection rate was higher among the male calves (38.46%) than females while the Rahaji breed had the highest prevalence of 62.5%. A total of 6 (18.18%) among the positive cases were diarrhoeic. The differences in prevalence based on sex, breeds and presence of diarrhoea were not statistically significant. Calves may become sources of Cryptosporidia infection to man and other animals in the study area through unrestricted movements and interactions with the environment.

  14. Prevalence of Cryptosporidium infection in camels (Camelus dromedarius) in a slaughterhouse in Iran.

    PubMed

    Razawi, S M; Oryan, A; Bahrami, S; Mohammadalipour, A; Gowhari, M

    2009-12-01

    Cryptosporidiosis is a zoonotic protozoan disease of worldwide distribution, affecting a wide range of vertebrate hosts. Most data on the biology, distribution pattern, pathology and prevalence of cryptosporidial infection in farm animals is restricted to cattle, sheep and goats. Limited data is available in other domestic herbivores including camel. Numerous camels (Camelus dromedarius) are raised in the semi-arid regions of Iran. Although camel is acknowledged as a potential source of contamination, little is known with regards to the prevalence of Cryptosporidium in this population except a case report on the occurrence of this infection in a bactrian camel (Camelus bactrianus) in China. This investigation was undertaken to determine the prevalence of Cryptosporidium infection in camels (C. dromedarius) from Najaf-Abad slaughterhouse, Isfahan Province, central part of Iran. Out of 103 faecal samples from 63 adult males and 40 adult females, 2-14 years old, 39 (37.9%) were found positive for oocysts. No significant differences were observed between males and females, and among different age groups. There was also no significant difference among infection intensity in different age groups. This is the first report of Cryptosporidium infection in camels from this country.

  15. 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

  16. 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

  17. 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.

  18. 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

  19. Utilizing HaloTag Technology to Track the Fate of PCSK9 from Intracellular vs. Extracellular Sources.

    PubMed

    Ai, Xi; Fischer, Paul; Palyha, Oksana C; Wisniewski, Douglas; Hubbard, Brian; Akinsanya, Karen; Strack, Alison M; Ehrhardt, Anka G

    2012-01-01

    The function of a particular protein is dependent upon its localization and milieu. The ability to track the "fate" of a protein is a valuable tool to elucidate its function. We present the use of HaloTag technology to study the localization and fate of human Proprotein Convertase Subtilisin-like Kexin type 9 (PCSK9).The role of PCSK9 in the regulation of circulating low density lipoprotein-cholesterol (LDL-c) levels is ascribed to binding of circulating PCSK9 to the LDL receptor (LDLR) and subsequent lysosomal degradation of LDLR. However, hints in the literature indicate that intracellular PCSK9 may act on the LDLR, possibly during processing of newly synthesized protein. To address this question, the source and fate of intracellular PCSK9 requires further investigation.We applied HaloTag technology to distinguish the source of intracellular PCSK9 and showed that newly synthesized intracellular PCSK9 has unique localization from the PCSK9 after re-uptake. This suggests different functions of PCSK9 while interacting with the LDLR.

  20. 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.

  1. 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. PMID:25270421

  2. Viral Multiplex Quantitative PCR Assays for Tracking Sources of Fecal Contamination▿

    PubMed Central

    Wolf, Sandro; Hewitt, Joanne; Greening, Gail E.

    2010-01-01

    Human and animal fecal pollution of the environment presents a risk to human health because of the presence of pathogenic viruses and bacteria. To distinguish between human and animal sources of pollution, we designed specific real-time reverse transcription (RT)-PCR assays for human and animal enteric viruses, including norovirus genogroups I, II, and III; porcine adenovirus types 3 and 5; ovine adenovirus; atadenovirus; and human adenovirus species C and F, which are excreted by infected humans, pigs, cattle, sheep, deer, and goats, and for the detection of F+ RNA bacteriophage genogroups I to IV, which are associated with human and animal wastes. The sensitivity of this viral toolbox (VTB) was tested against 10-fold dilution series of DNA plasmids that carry the target sequences of the respective viruses and was shown to detect at least 10 plasmid copies for each assay. A panel of human and animal enteric and respiratory viruses showed these assays to be highly sensitive and specific to their respective targets. The VTB was used to detect viruses in fecal and environmental samples, including raw sewage and biosolids from municipal sewage treatment plants, abattoir sewage, and fecally contaminated shellfish and river water, which were likely to contain animal or human viruses. PMID:20061455

  3. 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

  4. 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.

  5. Molecular Characterization of Cryptosporidium Isolates Obtained from Humans in France

    PubMed Central

    Guyot, K.; Follet-Dumoulin, A.; Lelièvre, E.; Sarfati, C.; Rabodonirina, M.; Nevez, G.; Cailliez, J. C.; Camus, D.; Dei-Cas, E.

    2001-01-01

    Cryptosporidium parvum is usually considered the agent of human cryptosporidiosis. However, only in the last few years, molecular biology-based methods have allowed the identification of Cryptosporidium species and genotypes, and only a few data are available from France. In the present work, we collected samples of whole feces from 57 patients from France (11 immunocompetent patients, 35 human immunodeficiency virus [HIV]-infected patients, 11 immunocompromised but non-HIV-infected patients) in whom Cryptosporidium oocysts were recognized by clinical laboratories. A fragment of the Cryptosporidium 18S rRNA gene encompassing the hypervariable region was amplified by PCR and sequenced. The results revealed that the majority of the patients were infected with cattle (29 of 57) or human (18 of 57) genotypes of Cryptosporidium parvum. However, a number of immunocompromised patients were infected with C. meleagridis (3 of 57), C. felis (6 of 57), or a new genotype of C. muris (1 of 57). This is the first report of the last three species of Cryptosporidium in humans in France. These results indicate that immunocompromised individuals are susceptible to a wide range of Cryptosporidium species and genotypes. PMID:11574558

  6. 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. PMID:27658593

  7. 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.

  8. Reduction of Cryptosporidium and Giardia by sewage treatment processes.

    PubMed

    Lim, Y A L; Wan Hafiz, W I; Nissapatorn, V

    2007-06-01

    Cryptosporidium and Giardia are two important pathogenic parasites that have caused many waterborne outbreaks which affected hundreds of thousands of people. Contamination from effluent discharged by sewage treatment plants have been implicated in previous waterborne outbreaks of Cryptosporidium and Giardia. This study evaluated the reduction of Cryptosporidium and Giardia (oo)cysts in two sewage treatment plants (STPA and STPB) in Malaysia which employed different treatment processes for a period of a year. Raw sewage influents and treated sewage effluents were concentrated by repeated centrifugation, subjected to sucrose density flotation and concentrated to a minimal volume depending upon the levels of contaminating debris. Cryptosporidium oocysts and Giardia cysts were enumerated using epifluorescence microscopy. The parasite concentrations in raw sewage were 18-8480 of Giardia cysts/litre and 1-80 of Cryptosporidium oocysts/litre. In treated sewage, the concentration of parasites ranged from 1-1462 cysts/litre and 20-80 oocysts/ litre for Giardia and Cryptosporidium respectively. Statistical analysis showed that sewage treatment process which employed extended aeration could reduce the concentration of Cryptosporidium and Giardia (oo)cysts significantly but treatment process which encompasses aerated lagoon could only reduce the concentration of Giardia cysts but not Cryptosporidium oocysts significantly. This phenomenon is of great concern in areas whereby effluent of sewage treatment plants is discharged into the upstream of rivers that are eventually used for abstraction of drinking water. Therefore, it is important that wastewater treatment authorities rethink the relevance of Cryptosporidium and Giardia contamination levels in wastewater and watersheds and to develop countermeasures in wastewater treatment plants. Further epidemiological studies on the occurrence and removal of pathogenic organisms from excreta and sewage are also recommended, in order

  9. Cryptosporidium and cryptosporidiosis: the African perspective.

    PubMed

    Aldeyarbi, Hebatalla M; Abu El-Ezz, Nadia M T; Karanis, Panagiotis

    2016-07-01

    The present overview discusses the findings of cryptosporidiosis research conducted in Africa and highlights the currently available information on Cryptosporidium epidemiology, genetic diversity, and distribution on the African continent, particularly among vulnerable populations, including children. It also emphasizes the burden of cryptosporidiosis, which is underestimated due to the presence of many silent asymptomatic carriers.Cryptosporidiosis is recognized as one of the leading causes of childhood diarrhea in African countries. It has dramatic adverse effects on child growth and development and causes increased mortality on a continent where HIV, poverty, and lack of sanitation and infrastructure increase the risk of cryptosporidial waterborne infection. PMID:27126869

  10. 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.

  11. 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

  12. Phylogenetic relationships of Cryptosporidium determined by ribosomal RNA sequence comparison.

    PubMed

    Johnson, A M; Fielke, R; Lumb, R; Baverstock, P R

    1990-04-01

    Reverse transcription of total cellular RNA was used to obtain a partial sequence of the small subunit ribosomal RNA of Cryptosporidium, a protist currently placed in the phylum Apicomplexa. The semi-conserved regions were aligned with homologous sequences in a range of other eukaryotes, and the evolutionary relationships of Cryptosporidium were determined by two different methods of phylogenetic analysis. The prokaryotes Escherichia coli and Halobacterium cuti were included as outgroups. The results do not show an especially close relationship of Cryptosporidium to other members of the phylum Apicomplexa. PMID:2332273

  13. Molecular characterization of Cryptosporidium isolates from humans in Equatorial Guinea.

    PubMed

    Blanco, María Alejandra; Iborra, Asunción; Vargas, Antonio; Nsie, Eugenia; Mbá, Luciano; Fuentes, Isabel

    2009-12-01

    The aim of the study was to perform a molecular characterization of clinical isolates of Cryptosporidium species from Equatorial Guinea. Standard laboratory methods were used to identify 35 cryptosporidiosis cases among 185 patients. PCR-RFLP successfully identified 34 Cryptosporidium species from these 35 cases, comprising C. parvum (52.9%), C. hominis (44.1%) and C. meleagridis (2.9%); over 90% of the species were isolated from HIV-positive patients. This is the first report of the molecular characterization of Cryptosporidium species isolated from humans in Equatorial Guinea and shows that zoonotic and anthroponotic transmission is present in this country.

  14. 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. PMID:27419102

  15. Enhancing Cryptosporidium parvum recovery rates for improved water monitoring.

    PubMed

    Pavli, Pagona; Venkateswaran, Sesha; Bradley, Mark; Bridle, Helen

    2016-01-01

    Water monitoring is essential to ensure safe drinking water for consumers. However existing methods have several drawbacks, particularly with regard to the poor recovery of Cryptosporidium due to the inability to efficiently elute Cryptosporidium oocysts during the established detection process used by water utilities. Thus the development of new inexpensive materials that could be incorporated into the concentration and release stage that would control Cryptosporidium oocysts adhesion would be beneficial. Here we describe improved filter performance following dip-coating of the filters with a "bioactive" polyacrylate. Specifically 69% more oocysts were eluted from the filter which had been coated with a polymer than on the naked filter alone.

  16. 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

  17. Identification of novel Cryptosporidium species in aquarium fish.

    PubMed

    Zanguee, N; Lymbery, J A; Lau, J; Suzuki, A; Yang, R; Ng, J; Ryan, U

    2010-11-24

    Little is known about the prevalence and genotypes of Cryptosporidium in fish. The present study investigated the prevalence of Cryptosporidium species in 200 aquarium fish of 39 different species in Western Australia by PCR amplification at the 18S rRNA locus. A total of 21 positives were detected by PCR (10.5% prevalence) from 13 different species of fish. Nineteen of these isolates were successfully sequenced. Of these, 12 were similar or identical to previously described species/genotypes of Cryptosporidium, while the remaining seven isolates appeared to represent three novel species.

  18. 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

  19. 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...

  20. 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...

  1. 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.

  2. 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.

  3. 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

  4. 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

  5. Cryptosporidium infection in solid organ transplantation.

    PubMed

    Florescu, Diana F; Sandkovsky, Uriel

    2016-09-24

    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

  6. Cryptosporidium hominis Is a Newly Recognized Pathogen in the Arctic Region of Nunavik, Canada: Molecular Characterization of an Outbreak

    PubMed Central

    Dixon, Brent; Dion, Réjean; Levesque, Benoît; Cantin, Philippe; Cédilotte, Lyne; Ndao, Momar; Proulx, Jean-François; Yansouni, Cedric P.

    2016-01-01

    Background Cryptosporidium is a leading cause of childhood diarrhea in low-resource settings, and has been repeatedly associated with impaired physical and cognitive development. In May 2013, an outbreak of diarrhea caused by Cryptosporidium hominis was identified in the Arctic region of Nunavik, Quebec. Human cryptosporidiosis transmission was previously unknown in this region, and very few previous studies have reported it elsewhere in the Arctic. We report clinical, molecular, and epidemiologic details of a multi-village Cryptosporidium outbreak in the Canadian Arctic. Methodology/Principal Findings We investigated the occurrence of cryptosporidiosis using a descriptive study of cases with onset between April 2013 and April 2014. Cases were defined as Nunavik inhabitants of any age presenting with diarrhea of any duration, in whom Cryptosporidium oocysts were detected by stool microscopy in a specialised reference laboratory. Cryptosporidium was identified in stool from 51 of 283 individuals. The overall annual incidence rate (IR) was 420 / 100,000 inhabitants. The IR was highest among children aged less than 5 years (1290 /100,000 persons). Genetic subtyping for stool specimens from 14/51 cases was determined by DNA sequence analysis of the 60 kDa glycoprotein (gp60) gene. Sequences aligned with C. hominis subtype Id in all cases. No common food or water source of infection was identified. Conclusions/Significance In this first observed outbreak of human cryptosporidiosis in this Arctic region, the high IR seen is cause for concern about the possible long-term effects on growth and development of children in Inuit communities, who face myriad other challenges such as overcrowding and food-insecurity. The temporal and geographic distribution of cases, as well as the identification of C. hominis subtype Id, suggest anthroponotic rather than zoonotic transmission. Barriers to timely diagnosis delayed the recognition of human cryptosporidiosis in this remote

  7. GazeParser: an open-source and multiplatform library for low-cost eye tracking and analysis.

    PubMed

    Sogo, Hiroyuki

    2013-09-01

    Eye movement analysis is an effective method for research on visual perception and cognition. However, recordings of eye movements present practical difficulties related to the cost of the recording devices and the programming of device controls for use in experiments. GazeParser is an open-source library for low-cost eye tracking and data analysis; it consists of a video-based eyetracker and libraries for data recording and analysis. The libraries are written in Python and can be used in conjunction with PsychoPy and VisionEgg experimental control libraries. Three eye movement experiments are reported on performance tests of GazeParser. These showed that the means and standard deviations for errors in sampling intervals were less than 1 ms. Spatial accuracy ranged from 0.7° to 1.2°, depending on participant. In gap/overlap tasks and antisaccade tasks, the latency and amplitude of the saccades detected by GazeParser agreed with those detected by a commercial eyetracker. These results showed that the GazeParser demonstrates adequate performance for use in psychological experiments.

  8. 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.

  9. 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 ...

  10. Detection of Cryptosporidium parvum in environmental soil and vegetables.

    PubMed

    Hong, Semie; Kim, Kyungjin; Yoon, Sejoung; Park, Woo-Yoon; Sim, Seobo; Yu, Jae-Ran

    2014-10-01

    Cryptosporidium parvum is a zoonotic protozoan parasite that causes cryptosporidial enteritis. Numerous outbreaks of cryptosporidiosis have been reported worldwide. Cryptosporidium is transmitted to hosts via consumption of contaminated water and food but also by direct contact with contaminated soil or infected hosts. The present study investigated farm soil collected from 34 locations along the western Korean peninsula and 24 vegetables purchased from local grocery markets in Seoul. The soil and vegetable samples were examined by real-time polymerase chain reaction (qPCR) to estimate the risk of infection. Eleven of 34 locations (32.4%) and 3 of 24 vegetable samples (12.5%) were contaminated with Cryptosporidium parvum, as confirmed by TaqI enzyme digestion of qPCR products and DNA sequencing. It is suggested that Cryptosporidium infection can be mediated via farm soil and vegetables. Therefore, it is necessary to reduce contamination of this organism in view of public health.

  11. Subtyping Cryptosporidium ubiquitum,a Zoonotic Pathogen Emerging in Humans

    PubMed Central

    Li, Na; Xiao, Lihua; Alderisio, Keri; Elwin, Kristin; Cebelinski, Elizabeth; Chalmers, Rachel; Santin, Monica; Fayer, Ronald; Kvac, Martin; Ryan, Una; Sak, Bohumil; Stanko, Michal; Guo, Yaqiong; Wang, Lin; Zhang, Longxian; Cai, Jinzhong; Roellig, Dawn

    2014-01-01

    Cryptosporidium ubiquitum is an emerging zoonotic pathogen. In the past, it was not possible to identify an association between cases of human and animal infection. We conducted a genomic survey of the species, developed a subtyping tool targeting the 60-kDa glycoprotein (gp60) gene, and identified 6 subtype families (XIIa–XIIf) of C. ubiquitum. Host adaptation was apparent at the gp60 locus; subtype XIIa was found in ruminants worldwide, subtype families XIIb–XIId were found in rodents in the United States, and XIIe and XIIf were found in rodents in the Slovak Republic. Humans in the United States were infected with isolates of subtypes XIIb–XIId, whereas those in other areas were infected primarily with subtype XIIa isolates. In addition, subtype families XIIb and XIId were detected in drinking source water in the United States. Contact with C. ubiquitum–infected sheep and drinking water contaminated by infected wildlife could be sources of human infections. PMID:24447504

  12. 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...

  13. 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...

  14. 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...

  15. 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...

  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. 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...

  18. Development of a Multilocus Sequence Tool for Typing Cryptosporidium muris and Cryptosporidium andersoni▿

    PubMed Central

    Feng, Yaoyu; Yang, Wenli; Ryan, Una; Zhang, Longxian; Kváč, Martin; Koudela, Břetislav; Modrý, David; Li, Na; Fayer, Ronald; Xiao, Lihua

    2011-01-01

    Although widely used for the characterization of the transmission of intestinal Cryptosporidium spp., genotyping tools are not available for C. muris and C. andersoni, two of the most common gastric Cryptosporidium spp. infecting mammals. In this study, we screened the C. muris whole-genome sequencing data for microsatellite and minisatellite sequences. Among the 13 potential loci (6 microsatellite and 7 minisatellite loci) evaluated by PCR and DNA sequencing, 4 were eventually chosen. DNA sequence analyses of 27 C. muris and 17 C. andersoni DNA preparations showed the presence of 5 to 10 subtypes of C. muris and 1 to 4 subtypes of C. andersoni at each locus. Altogether, 11 C. muris and 7 C. andersoni multilocus sequence typing (MLST) subtypes were detected among the 16 C. muris and 12 C. andersoni specimens successfully sequenced at all four loci. In all analyses, the C. muris isolate (TS03) that originated from an East African mole rat differed significantly from other C. muris isolates, approaching the extent of genetic differences between C. muris and C. andersoni. Thus, an MLST technique was developed for the high-resolution typing of C. muris and C. andersoni. It should be useful for the characterization of the population genetics and transmission of gastric Cryptosporidium spp. PMID:20980577

  19. Molecular characterisation of Cryptosporidium isolates from pet reptiles.

    PubMed

    Pedraza-Díaz, Susana; Ortega-Mora, Luis M; Carrión, Beatriz A; Navarro, Vanesa; Gómez-Bautista, Mercedes

    2009-03-23

    In this study, we investigated the presence of Cryptosporidium in 171 faecal samples from reptiles commonly used as pet animals. These include lizards belonging to the genera Eublepharis, Pogona, Chlamydosaurus, Hemiteconyx, Teratoscincus, Tiliqua, Iguana, and Chamaeleo, snakes of the genera Lampropeltis, Elaphe, Python, Boa and Corallus, and tortoises belonging to the genera Testudo and Kinixys. Cryptosporidium oocysts were detected by immunofluorescence using a commercially available kit and cryptosporidial DNA by amplification of a polymorphic fragment of the 18S rDNA and the HSP70 locus. Cryptosporidium was detected in 38.6% and 25.1% of the samples analysed by immunofluorescence and PCR, respectively. Molecular characterisation of the isolates confirmed that C. serpentis and C. varanii (syn. C. saurophilum) are the main species involved in infection in pet reptiles but also showed the presence of C. parvum and C. muris, as well as other species or genotypes of this parasite including the Cryptosporidium mouse genotype and Cryptosporidium tortoise genotype previously described in reptiles. In addition, a Cryptosporidium sp. was isolated from a chameleon and a python.

  20. Non-coding RNAs in epithelial immunity to Cryptosporidium infection

    PubMed Central

    Zhou, Rui; Feng, Yaoyu; Chen, Xian-Ming

    2015-01-01

    SUMMARY Cryptosporidium spp. is a protozoan parasite that infects the gastrointestinal epithelium and causes diarrhoeal disease worldwide. It is one of the most common pathogens responsible for moderate to severe diarrhoea in children younger than 2 years. Because of the ‘minimally invasive’ nature of Cryptosporidium infection, mucosal epithelial cells are critical to the host’s anti-Cryptosporidium immunity. Gastrointestinal epithelial cells not only provide the first and most rapid defence against Cryptosporidium infection, they also mobilize immune effector cells to the infection site to activate adaptive immunity. Recent advances in genomic research have revealed the existence of a large number of non-protein-coding RNA transcripts, so called non-coding RNAs (ncRNAs), in mammalian cells. Some ncRNAs may be key regulators for diverse biological functions, including innate immune responses. Specifically, ncRNAs may modulate epithelial immune responses at every step of the innate immune network following Cryptosporidium infection, including production of antimicrobial molecules, expression of cytokines/chemokines, release of epithelial cell-derived exosomes, and feedback regulation of immune homoeostasis. This review briefly summarizes the current science on ncRNA regulation of innate immunity to Cryptosporidium, with a focus on microRNA-associated epithelial immune responses. PMID:24828969

  1. Study on Cryptosporidium contamination in vegetable farms around Tehran.

    PubMed

    Ranjbar-Bahadori, Sh; Mostoophi, A; Shemshadi, B

    2013-06-01

    In recent years, an increase in the number of cases of food-borne illnesses linked to fresh vegetables has been reported. One of the causative agents of these infections is Cryptosporidium and it appears that one route of transmission to humans is food-borne, so fruits and vegetables have important roles. The goal of this study was to determine the level of Cryptosporidium contamination in vegetable farms around Tehran, Iran. A total of 496 samples from 115 vegetable farms in different regions around Tehran (Capital city of Iran) were collected and different types of vegetables were investigated for the parasite in June and July, 2012. A sediment concentration method followed by modified Ziehl-Neelsen's acid-fast staining was used to determine the presence of Cryptosporidium oocysts. Our findings revealed that 6.6% of studied samples were contaminated with Cryptosporidium species. The highest rate of contamination was reported in Bagher Abad (South of Tehran) (11.1%), and green onions were more commonly contaminated (14.8%) than any other vegetables tested. Furthermore, when waste water was used to irrigate vegetable farms, the contamination rate was (33.3%). Statistical analysis showed a correlation between contamination with Cryptosporidium spp. and studied risk factors including: different regions around Tehran, type of vegetables, and type of water used for farm irrigation. Therefore, vegetables may provide a route by which Cryptosporidium can be transmitted to humans, and control strategies should be considered. PMID:23959484

  2. Detection of Cryptosporidium hominis and novel Cryptosporidium bat genotypes in wild and captive Pteropus hosts in Australia.

    PubMed

    Schiller, Sabine Eva; Webster, Koa Narelle; Power, Michelle

    2016-10-01

    Spillover of zoonotic pathogens from wildlife to humans has been identified as a primary threat to global health. In contrast, the process of reverse pathogen transmission (zooanthroponosis), whereby pathogens move from humans into wildlife species is still largely unexplored. Globally, increasing urbanisation and habitat loss are driving many wildlife species into urban and regional centres. In Australia, large numbers of flying foxes now live in close proximity to humans, increasing the risk of zooanthroponosis. The protozoan parasite Cryptosporidium is an emerging zoonotic parasite that infects a wide range of vertebrates yet there are limited studies on transmission potential of Cryptosporidium between humans and urban wildlife. To explore the presence of zooanthroponosis in flying foxes in Australia the occurrence and diversity of Cryptosporidium was investigated in urbanised wild and captive flying foxes. PCR screening of faecal samples (n=281) from seven wild sites and two captive facilities identified the presence of Cryptosporidium in 3.2% (95% CI 1.5% to 6.0%) of faecal samples. In faecal samples from wild sites Cryptosporidium occurrence was 1.7% (95% CI 0.3% to 4.8%) versus 5.9% (95% CI 2.2% to 12.4%) in samples from captive individuals, with no significant difference between captive and wild sites (p=0.077). Multilocus sequencing using three loci, 18s rDNA, actin and gp60 was used to identify Cryptosporidium in flying fox species. The host specific Cryptosporidium hominis was identified in faecal samples from two captive flying foxes, and one of these samples was confirmed as C. hominis at both actin and gp60. Sequencing of the 18s rDNA also revealed four novel Cryptosporidium genotypes in wild and captive individuals, actin and gp60 amplification and sequencing were unreliable for all four novel genotypes. These novel genotypes have been designated Cryptosporidium bat genotypes VIII-XI. This first report of Cryptosporidium spp. in Australian flying

  3. Detection of Cryptosporidium hominis and novel Cryptosporidium bat genotypes in wild and captive Pteropus hosts in Australia.

    PubMed

    Schiller, Sabine Eva; Webster, Koa Narelle; Power, Michelle

    2016-10-01

    Spillover of zoonotic pathogens from wildlife to humans has been identified as a primary threat to global health. In contrast, the process of reverse pathogen transmission (zooanthroponosis), whereby pathogens move from humans into wildlife species is still largely unexplored. Globally, increasing urbanisation and habitat loss are driving many wildlife species into urban and regional centres. In Australia, large numbers of flying foxes now live in close proximity to humans, increasing the risk of zooanthroponosis. The protozoan parasite Cryptosporidium is an emerging zoonotic parasite that infects a wide range of vertebrates yet there are limited studies on transmission potential of Cryptosporidium between humans and urban wildlife. To explore the presence of zooanthroponosis in flying foxes in Australia the occurrence and diversity of Cryptosporidium was investigated in urbanised wild and captive flying foxes. PCR screening of faecal samples (n=281) from seven wild sites and two captive facilities identified the presence of Cryptosporidium in 3.2% (95% CI 1.5% to 6.0%) of faecal samples. In faecal samples from wild sites Cryptosporidium occurrence was 1.7% (95% CI 0.3% to 4.8%) versus 5.9% (95% CI 2.2% to 12.4%) in samples from captive individuals, with no significant difference between captive and wild sites (p=0.077). Multilocus sequencing using three loci, 18s rDNA, actin and gp60 was used to identify Cryptosporidium in flying fox species. The host specific Cryptosporidium hominis was identified in faecal samples from two captive flying foxes, and one of these samples was confirmed as C. hominis at both actin and gp60. Sequencing of the 18s rDNA also revealed four novel Cryptosporidium genotypes in wild and captive individuals, actin and gp60 amplification and sequencing were unreliable for all four novel genotypes. These novel genotypes have been designated Cryptosporidium bat genotypes VIII-XI. This first report of Cryptosporidium spp. in Australian flying

  4. Molecular characterization of Cryptosporidium spp. in domestic pigeons (Columba livia domestica) in Guangdong Province, Southern China.

    PubMed

    Li, Juan; Lin, Xuhui; Zhang, Longxian; Qi, Nanshan; Liao, Shenquan; Lv, Minna; Wu, Caiyan; Sun, Mingfei

    2015-06-01

    To investigate the prevalence and assess the zoonotic transmission burden of Cryptosporidium species in domestic pigeons in Guangdong Province, Southern China, 244 fecal samples were collected from four pigeon breeding farms between June 2012 and March 2013. Cryptosporidium oocysts were purified by Sheather's sugar flotation technique and characterized by DNA sequencing of small subunit ribosomal RNA (SSU rRNA) gene. Cryptosporidium species were determined by comparison of sequences with corresponding Cryptosporidium sequences in GenBank and phylogenetic analysis using neighbor-joining (NJ) in MEGA5.2. The overall prevalence of Cryptosporidium infection in domestic pigeons in Guangdong Province was 0.82% (2/244). Two Cryptosporidium species, namely Cryptosporidium baileyi and Cryptosporidium meleagridis, were identified in Huizhou and Chaozhou farm, respectively. These findings confirmed the existence of C. meleagridis infection in domestic pigeons in China for the first time and provided base-line information for further studies to evaluate the public health risk from pigeon to human.

  5. 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

  6. Decay of fecal indicator bacterial populations and bovine-associated source-tracking markers in freshly deposited cow pats.

    PubMed

    Oladeinde, Adelumola; Bohrmann, Thomas; Wong, Kelvin; Purucker, S T; Bradshaw, Ken; Brown, Reid; Snyder, Blake; Molina, Marirosa

    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.

  7. 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.

  8. 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.

  9. Prospects for immunotherapy and vaccines against Cryptosporidium

    PubMed Central

    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. PMID:24638018

  10. 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.

  11. Cryptosporidium species a "new" human pathogen.

    PubMed Central

    Casemore, D P; Sands, R L; Curry, A

    1985-01-01

    Publications describing aspects of the coccidian protozoan parasite Cryptosporidium, increased greatly during 1983 and 1984 as a result of not only increasing veterinary interest but also in the role of the parasite in the newly recognised acquired immune deficiency syndrome (AIDS). The reports reflected widespread collaboration, not only between clinicians, microbiologists, and histopathologists, but also between veterinary and human health care workers. Cryptosporidium was first described in mice in 1907 and subsequently in various other species; it was not described in man until 1976. Several likely putative species have been described, but there is probably little host specificity. Experimental and clinical studies have greatly increased the knowledge about the organism's biology. The parasite undergoes its complete life cycle within the intestine, although it may occasionally occur in other sites. The main symptom produced is a non-inflammatory diarrhoea, which, in patients with AIDS and children in Third World countries, may be life threatening: even in immunocompetent subjects this symptom is usually protracted. Attempts to find effective chemotherapeutic agents have been unsuccessful. Epidemiologically the infection was thought to be zoonotic in origin, but there is increasing evidence of person to person transmission. Diagnosis has depended upon histological examination, but simple methods of detection have now been described: more invasive methods need no longer be used. The parasite, which is found more commonly in children, occurs in about 2% of faecal specimens examined and seems to be closely associated with production of symptoms. A serological response has been shown. Much remains to be learned about its epidemiology and pathogenic mechanisms, while the expected increase in incidence of AIDS makes an effective form of treatment essential. Images PMID:3908490

  12. 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.

  13. 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. PMID:23810821

  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. Characterization of the Oum Er Rbia (Morocco) high basin karstic water sources by using solid state nuclear track detectors and radon as a natural tracer.

    PubMed

    Khalil, N; Misdaq, M A; Berrazzouk, S; Mania, J

    2002-06-01

    Uranium and thorium contents as well as radon alpha-activities per unit volume were evaluated inside different water samples by using a method based on calculating the CR-39 and LR-115 type II solid state nuclear track detectors (SSNTDs) detection efficiencies for the emitted alpha-particles and measuring the resulting track density rates. The validity of the SSNTD technique utilized was checked by analysing uranyl nitrate (UO2(NO3)26H2O) standard solutions. A relationship between water radon concentration and water transmission of different water sources belonging to two regions of the Middle Atlas (Morocco) water reservoir was found. The influence of the water flow rate as well as the permeability and fracture system of the host rocks of the sources studied was investigated.

  16. 40 CFR 141.716 - Source toolbox components.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... contamination to the system's source water. (iv) A statement of goals and specific actions the system will... Section 141.716 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium...

  17. 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. PMID:25687523

  18. 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.

  19. 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.

  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. 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

  2. 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

  3. Childhood Cryptosporidium infection among aboriginal communities in Peninsular Malaysia.

    PubMed

    Al-Mekhlafi, H M; Mahdy, M A K; 'Azlin, M Y; Fatmah, M S; Norhayati, M

    2011-03-01

    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

  4. Childhood Cryptosporidium infection among aboriginal communities in Peninsular Malaysia.

    PubMed

    Al-Mekhlafi, H M; Mahdy, M A K; 'Azlin, M Y; Fatmah, M S; Norhayati, M

    2011-03-01

    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.

  5. Identification of novel Cryptosporidium genotypes in kangaroos from Western Australia.

    PubMed

    Yang, Rongchang; Fenwick, Stanley; Potter, Abbey; Ng, Josephine; Ryan, Una

    2011-06-30

    A total of 763 faecal samples were collected from western grey kangaroos (Macropus fuliginosus) in Western Australia and screened for the presence of Cryptosporidium by PCR at the 18S ribosomal RNA (rRNA) locus. Samples that were positive at the 18S locus were also amplified at the actin locus. The overall prevalence was 9.3% (71/763). At the 18S rRNA locus, sequences were obtained for 28 of the 71 positives. Sequence analysis identified four species; Cryptosporidium fayeri in seven isolates, Cryptosporidium marcopodum in four isolates, Cryptosporidium xiaoi in six isolates and a novel genotype (kangaroo genotype I) in eleven isolates. Analysis at the actin locus confirmed the genetic distinctness of the novel genotype. The results of the present study indicate that in addition to C. fayeri and C. marcopodum, kangaroos may be capable of being infected with a wider range of Cryptosporidium species and genotypes including livestock species such as C. xiaoi. The novel genotype identified in the kangaroos most likely represents a cryptic species that requires further analyses to confirm its species status.

  6. Natural infection of Cryptosporidium muris in ostriches (Struthio camelus).

    PubMed

    Qi, Meng; Huang, Lei; Wang, Rongjun; Xiao, Lihua; Xu, Lina; Li, Junqiang; Zhang, Longxian

    2014-10-15

    A total of 303 fecal samples were collected from ostriches (Struthio camelus) and 31 samples (10.2%) were Cryptosporidium-positive upon microscopic analysis. The infection rate was 27.6% in ostriches aged 16-60 days, 1.2% in those aged 61-180 days, and 20.4% in those aged >10 years. The Cryptosporidium-positive isolates were genotyped with a restriction fragment length polymorphism analysis and DNA sequence analysis of the small subunit (SSU) rRNA gene. The 22 isolates from ostriches aged >10 years were identified as Cryptosporidium muris, whereas the nine isolates from ostriches <180 days were Cryptosporidium baileyi. Ten of the 22 C. muris isolates were analyzed based on the actin and HSP70 genes, and the results were identical to those observed for the SSU rRNA gene. Cross-transmission studies demonstrated that the C. muris isolate infected BALB/c mice and Mongolian gerbils, but did not infect chickens. C. muris isolated in this study appears to be host-adapted, consistent with a previous multilocus sequence typing analysis. Further studies are required to understand the prevalence and transmission of Cryptosporidium spp. in ostriches in different geographic areas. PMID:25178556

  7. 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. PMID:26986498

  8. 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.

  9. Distribution of Cryptosporidium species in Tibetan sheep and yaks in Qinghai, China.

    PubMed

    Li, Pei; Cai, Jinzhong; Cai, Min; Wu, Wenxian; Li, Chunhua; Lei, Mengtong; Xu, Hailing; Feng, Lijun; Ma, Jiawen; Feng, Yaoyu; Xiao, Lihua

    2016-01-15

    Few data are available on the distribution of Cryptosporidium species in Tibetan sheep and yaks, which are free-range animals living in a cold, low oxygen, and high ultraviolet radiation habitat. In this study, 904 fecal specimens were collected from 350 Tibetan sheep and 554 yaks in six counties. Cryptosporidium spp. were detected and differentiated by PCR and sequence analyses. Altogether, 43 (12.3%) Tibetan sheep and 158 (28.5%) yaks were positive for Cryptosporidium spp. In Tibetan sheep, Cryptosporidium xiaoi (39/43, 90.7%) was the dominant species, with the remaining cases (4/43, 9.3%) by Cryptosporidium ubiquitum. All C. ubiquitum specimens belonged to the subtype family XIIa. In contrast, Cryptosporidium andersoni (72/158, 45.6%), Cryptosporidium bovis (47/158, 29.7%), Cryptosporidium ryanae cattle type (35/158, 22.2%), C. ryanae buffalo type (2/158, 1.3%), and Cryptosporidium suis-like (2/158, 1.3%) were identified in yaks. Contradictory to previous observations, C. andersoni was one of the dominant Cryptosporidium species in yaks in this study. Despite sharing habitats, Tibetan sheep and yaks are evidently infected with different Cryptosporidium species.

  10. 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. PMID:26240518

  11. VideoHacking: Automated Tracking and Quantification of Locomotor Behavior with Open Source Software and Off-the-Shelf Video Equipment

    PubMed Central

    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. PMID:26240518

  12. 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.

  13. 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.

  14. Cryptosporidium muris in a reticulated giraffe (Giraffa camelopardalis reticulata).

    PubMed

    Kodádková, A; Kvác, M; Ditrich, O; Sak, B; Xiao, L

    2010-02-01

    Cryptosporidium spp. infection in captive exotic mammals was investigated using staining and molecular biological methods. A total of 323 fecal samples from 100 mammalian species (62 Artiodactyla, 33 Rodentia, 3 Perissodactyla, and 2 Paenungultata) in 4 zoological gardens in the Czech Republic was examined. Only in a reticulated giraffe (Giraffa camelopardalis reticulata) sample was Cryptosporidium sp. infection detected. The partial small subunit rRNA sequence obtained from the isolate was identical to sequences of Cryptosporidium muris in rock hyrax (Procavia capensis) and Bactrian camel (Camelus bactrianus). Neonatal BALB/c mice inoculated with 1 x 10(3) fresh oocysts of the C. muris giraffe isolate did not produce a detectable infection. PMID:19685941

  15. 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. PMID:23021408

  16. Waterborne Giardia cysts and Cryptosporidium oocysts in the Yukon, Canada.

    PubMed Central

    Roach, P D; Olson, M E; Whitley, G; Wallis, P M

    1993-01-01

    Several outbreaks of waterborne giardiasis have occurred in southern Canada, but nothing has been reported from the Canadian North. The objective of this study was to collect information relevant to waterborne giardiasis and cryptosporidiosis in the Yukon including epidemiological data and analyses of water, sewage, and animal fecal samples. Remote, pristine water samples were found to be contaminated with Giardia cysts (7 of 22 or 32%) but not with Cryptosporidium oocysts. Giardia cysts were found in 21% (13 of 61) of animal scats, but no Cryptosporidium oocysts were observed (small sample size). Whitehorse's drinking water was episodically contaminated with Giardia cysts (7 of 42 or 17%) and Cryptosporidium oocysts (2 of 42 or 5%). Neither were found in Dawson City's water supply. The only water treatment in the Yukon is chlorination, but contact times and free chlorine residuals are often too low to provide adequate protection by disinfection. Raw sewage samples from the five largest population centers in the Yukon contained 26 to 3,022 Giardia cysts and 0 to 74 Cryptosporidium oocysts per liter. Treated sewage from Whitehorse contained fewer Giardia cysts but more Cryptosporidium oocysts on average. Both were detected in Lake Laberge, downstream of Whitehorse, which has a history of fecal coliform contamination. Daily monitoring of raw sewage from the suburbs of Whitehorse showed a summertime peak of Giardia cysts and occasional Cryptosporidium oocysts after springtime contamination of drinking water. Despite this evidence, epidemiological data for the Yukon showed an endemic infection rate of only 0.1% for giardiasis (cryptosporidiosis is not notifiable).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8439168

  17. Serological responses to Cryptosporidium antigens among women using riverbank-filtered water, conventionally filtered surface water and groundwater in Hungary.

    PubMed

    Frost, Floyd; Craun, Gunther; Mihály, Kádár; György, Berencsi; Calderon, Rebecca; Muller, Tm

    2005-03-01

    We compared serological responses to Cryptosporidium parvum antigens using surplus sera from females undergoing routine screening for pregnancy from three counties in Hungary where bank-filtered surface water, conventionally filtered and disinfected surface water, and groundwater from either a karst or confined aquifer are commonly used for drinking water. The primary purpose was to determine whether the prevalence and intensity of serological responses, indicators of prior Cryptosporidium infection were similar for these populations. Women using groundwater from a confined aquifer had significantly lower mean serological responses for both the 15/17-kDa and 27-kDa (p < 0.0001) antigen groups than women using conventionally filtered and disinfected surface water or karst well water. This is suggestive of less frequent infections. Women using bank-filtered water also had lower mean responses for both antigen groups. Among women using bank-filtered water, the mean intensity of response for both antigen groups was almost one-third of the mean response observed for women using conventionally filtered and disinfected surface water. These findings suggest that riverbank filtration may be an effective alternative to conventional treatment for reducing Cryptosporidium exposures and infection from surface drinking water sources.

  18. 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

  19. 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.

  20. Viability of preserved Cryptosporidium baileyi oocysts

    PubMed Central

    Surl, Chan-Gu; Kim, Se-Min

    2003-01-01

    The present study was undertaken to determine the viability and infectivity of oocysts of Cryptosporidium baileyi that had been stored from 1 to 40 months at 4℃ preserved in 2.5% potassium dichromate solution. Oocysts of C. baileyi were purified from the feces of experimentally infected chickens using discontinuous sucrose gradients. Subsequently, the purified oocysts were suspended in 2.5% potassium dichromate solution at a concentration of 1 × 107 organism/ml, and their viabilities were assessed by nucleic acid staining, histologic examination, and infectivity to 2-day-old chickens. All chickens inoculated with oocysts that had been stored for 1-18 months developed patent infections, while chickens infected with older oocysts remained uninfected. Between 5.8% and 82.2% of the oocysts, stored at 4℃ in 2.5% potassium dichromate solution, were found to be viable, as determined by nucleic acid staining. Parasite colonization in the bursa of Fabricius was detected in the microvillus border of bursal epithelium. The finding that C. baileyi oocysts remain infective to chickens for at least 18 months offers important time-saving advantages to investigators who frequently require large numbers of oocysts. PMID:14699260

  1. 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

  2. Detection of Cyclospora, Cryptosporidium, and Giardia in ready-to-eat packaged leafy greens in Ontario, Canada.

    PubMed

    Dixon, Brent; Parrington, Lorna; Cook, Angela; Pollari, Frank; Farber, Jeffrey

    2013-02-01

    Numerous foodborne outbreaks of diarrheal illness associated with the consumption of produce contaminated with protozoan parasites have been reported in North America in recent years. The present study reports on the presence of Cyclospora, Cryptosporidium, and Giardia in precut salads and leafy greens purchased at retail in Ontario, Canada. A total of 544 retail samples were collected between April 2009 and March 2010 and included a variety of salad blends and individual leafy greens. Most of these products were grown in the United States, with some from Canada and Mexico. Parasites were eluted and concentrated before detection by PCR and immunofluorescence microscopy. DNA sequences were aligned with reference sequences in GenBank. Cyclospora spp. were identified by PCR-restriction fragment length polymorphism in nine (1.7 % ) samples and by DNA sequence analysis. Cryptosporidium spp. were identified in 32 (5.9%) samples; 29 were sequenced and aligned with the zoonotic species Cryptosporidium parvum. Giardia duodenalis was identified in 10 (1.8%) samples, and of the 9 samples successfully sequenced, 7 aligned with G. duodenalis assemblage B and 2 with assemblage A, both of which are also zoonotic. The presence of Cryptosporidium oocysts and Giardia cysts was confirmed in some of the PCR-positive samples using microscopy, while Cyclospora -like oocysts were observed in most of the Cyclospora PCR-positive samples. The relatively high prevalence of these parasites in packaged salads and leafy greens establishes a baseline for further studies and suggests a need for additional research with respect to the possible sources of contamination of these foods, the determination of parasite viability and virulence, and means to reduce foodborne transmission to humans. PMID:23433379

  3. 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.

  4. Multi-scale Cryptosporidium/sand interactions in water treatment.

    PubMed

    Tufenkji, Nathalie; Dixon, David R; Considine, Robert; Drummond, Calum J

    2006-10-01

    Owing to its widespread occurrence in drinking water supplies and its significant resistance to environmental stresses, Cryptosporidium parvum is regarded as one of the most important waterborne microbial parasites. Accordingly, a substantial research effort has been aimed at elucidating the physical, chemical and biological factors controlling the transport and removal of Cryptosporidium oocysts in natural subsurface environments and drinking water treatment facilities. In this review, a multi-scale approach is taken to discuss the current state-of-knowledge on Cryptosporidium-sand interactions at a nano-scale, bench-scale and field-scale relevant to water treatment operations. Studies conducted at the nano-scale and bench-scale illustrate how techniques based on the principles of colloid and surface chemistry are providing new insights about oocyst-sand interactions during transport of Cryptosporidium oocysts in granular porous media. Specifically, atomic force microscopy and impinging jet experiments reveal the importance of oocyst surface biomolecules in controlling Cryptosporidium/sand interactions by a mechanism of steric hindrance. Traditional bench-scale column transport studies conducted over a broad range of experimental conditions highlight the role of physicochemical filtration and physical straining in the removal of oocysts from the pore fluid. Such experiments have also been used to evaluate the influence of biofilms formed on grain surfaces and the presence of natural organic matter on oocyst-sand interactions. Whilst filtration studies conducted at the plant-scale have been useful for evaluating the effectiveness of various materials as surrogates for Cryptosporidium oocysts, at this macro-scale, little could be learnt about the fundamental mechanisms controlling oocyst-sand interactions. This review of the literature on Cryptosporidium-sand interactions at different length scales points to the importance of combining studies at the plant

  5. 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.

  6. Tracking strategies involving fourteen sources for locating a transient study sample: parents of sudden infant death syndrome infants and control infants.

    PubMed

    Klonoff-Cohen, H

    1996-07-01

    Strategies involving 14 sources were used to locate 230 parents of sudden infant death syndrome infants who died in Southern California between 1989 and 1992 and 255 parents of healthy, living infants matched by birth hospital, birth date, race and sex. The sample consisted of adults of reproductive age residing in Southern California. After an event of sudden infant death, many parents moved without a forwarding address; only their names, last known address, and the infant's race, birth date, and sex were available. There was no access to birth certificates, obstetric or pediatric medical records, parents' Social Security numbers, or parents' birth dates. The most successful tracking sources for case parents were the Department of Motor Vehicles, postal service, reverse directory and neighbors, private investigator, and California Medicaid services. For control parents, the post office, Department of Motor Vehicles, and Folks Finders proved the most helpful. Using a combination of the 14 sources achieved an adequate sample size.

  7. 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. PMID:25281384

  8. 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.

  9. 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

  10. 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.

  11. 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. PMID:26437239

  12. Complex epidemiology and zoonotic potential for Cryptosporidium suis in rural Madagascar.

    PubMed

    Bodager, Jonathan R; Parsons, Michele B; Wright, Patricia C; Rasambainarivo, Fidisoa; Roellig, Dawn; Xiao, Lihua; Gillespie, Thomas R

    2015-01-15

    Cryptosporidium spp. is the most important parasitic diarrheal agent in the world, is among the top four causes of moderate-to-severe diarrheal disease in young children in developing nations, and is problematic as an opportunistic co-infection with HIV. In addition, Cryptosporidium is a persistent challenge for livestock production. Despite its zoonotic potential, few studies have examined the ecology and epidemiology of this pathogen in rural systems characterized by high rates of overlap among humans, domesticated animals, and wildlife. To improve our understanding of the zoonotic potential of Cryptosporidium species in the rural tropics, we screened humans, livestock, peridomestic rodents, and wildlife using PCR-RFLP and sequencing-based approaches to distinguish species of Cryptosporidium in rural southeastern Madagascar. Cryptosporidium of multiple species/genotypes were apparent in this study system. Interestingly, C. suis was the dominant species of Cryptosporidium in the region, infecting humans (n=1), cattle (n=18), pigs (n=3), and rodents (n=1). The broad species range of C. suis and the lack of common cattle Cryptosporidium species (Cryptosporidium parvum and Cryptosporidium andersoni) in this system are unique. This report represents the fifth confirmed case of C. suis infection in humans, and the first case in Africa. Few rural human and livestock populations have been screened for Cryptosporidium using genus-specific genotyping methods. Consequently, C. suis may be more widespread in human and cattle populations than previously believed.

  13. 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.

  14. Assessment of polymorphic genetic markers for multi-locus typing of Cryptosporidium parvum and Cryptosporidium hominis.

    PubMed

    Robinson, Guy; Chalmers, Rachel M

    2012-10-01

    The use of high resolution molecular tools to study Cryptosporidium parvum and Cryptosporidium hominis intra-species variation is becoming common practice, but there is currently no consensus in the methods used. The most commonly applied tool is partial gp60 gene sequence analysis. However, multi-locus schemes are acknowledged to improve resolution over analysis of a single locus, which neglects potential re-assortment of genes during the sexual phase of the Cryptosporidium life-cycle. Multi-locus markers have been investigated in isolates from a variety of sampling frames, in varying combinations and using different assays and methods of analysis. To identify the most informative markers as candidates for the development of a standardised multi-locus fragment size-based typing (MLFT) scheme to integrate with epidemiological analyses, we examined the published literature. A total of 31 MLFT studies were found, employing 55 markers of which 45 were applied to both C. parvum and C. hominis. Of the studies, 11 had sufficient raw data, from three or more markers, and a sampling frame containing at least 50 samples, for meaningful in-depth analysis using assessment criteria based on the sampling frame, study size, number of markers investigated in each study, marker characteristics (>2 nucleotide repeats) and the combinations of markers generating all possible multi-locus genotypes. Markers investigated differed between C. hominis and C. parvum. When each scheme was analysed for the fewest markers required to identify 95% of all MLFTs, some redundancy was identified in all schemes; an average redundancy of 40% for C. hominis and 27% for C. parvum. Ranking markers, based on the most productive combinations, identified two different sets of potentially most informative candidate markers, one for each species. These will be subjected to technical evaluation including typability (percentage of samples generating a complete multi-locus type) and discriminatory power by

  15. 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.

  16. Genetic modification of the diarrheal pathogen Cryptosporidium parvum

    PubMed Central

    Vinayak, Sumiti; Pawlowic, Mattie C.; Sateriale, Adam; Brooks, Carrie F.; Studstill, Caleb J.; Bar-Peled, Yael; Cipriano, Michael J.; Striepen, Boris

    2015-01-01

    Recent studies into the global causes of severe diarrhea in young children have identified the protozoan parasite Cryptosporidium as the second most important diarrheal pathogen after rotavirus1–3. Diarrheal disease is estimated to be responsible for 10.5% of overall child mortality4. Cryptosporidium is also an opportunistic pathogen in the context of HIV-AIDS and organ transplantation5,6. There is no vaccine and only a single approved drug that provides no benefit for those in gravest danger, malnourished children and immunocompromised patients7,8. Cryptosporidiosis drug and vaccine development is limited by the poor tractability of the parasite, which includes lack of continuous culture, facile animal models, and molecular genetic tools3,9. Here we describe an experimental framework to genetically modify this important human pathogen. We establish and optimize transfection of C. parvum sporozoites in tissue culture. To isolate stable transgenics we develop a mouse model that delivers sporozoites directly into the intestine, a Cryptosporidium CRISPR/Cas9 system, and in vivo selection for aminoglycoside resistance. We derive reporter parasites suitable for in vitro and in vivo drug screening, and we evaluate the basis of drug susceptibility by gene knock out. We anticipate the ability to genetically engineer the 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. PMID:26176919

  17. 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

  18. 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...

  19. 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...

  20. 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 ...

  1. 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...

  2. 40 CFR 141.712 - Unfiltered system Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 24 2013-07-01 2013-07-01 false Unfiltered system Cryptosporidium treatment requirements. 141.712 Section 141.712 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY...)(ii) are in violation of the treatment technique requirement. (d) Use of two disinfectants....

  3. 40 CFR 141.712 - Unfiltered system Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 23 2014-07-01 2014-07-01 false Unfiltered system Cryptosporidium treatment requirements. 141.712 Section 141.712 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY...)(ii) are in violation of the treatment technique requirement. (d) Use of two disinfectants....

  4. 40 CFR 141.712 - Unfiltered system Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 24 2012-07-01 2012-07-01 false Unfiltered system Cryptosporidium treatment requirements. 141.712 Section 141.712 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY...)(ii) are in violation of the treatment technique requirement. (d) Use of two disinfectants....

  5. 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...

  6. 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...

  7. Biofilms Reduce Solar Disinfection of Cryptosporidium parvum Oocysts

    PubMed Central

    Hargreaves, B. R.; Jellison, K. L.

    2012-01-01

    Solar radiation reduces Cryptosporidium infectivity. Biofilms grown from stream microbial assemblages inoculated with oocysts were exposed to solar radiation. The infectivity of oocysts attached at the biofilm surface and oocysts suspended in water was about half that of oocysts attached at the base of a 32-μm biofilm. PMID:22467508

  8. DIFFERENTIATING HUMAN FROM ANIMAL ISOLATES OF CRYPTOSPORIDIUM PARVUM

    EPA Science Inventory

    We analyzed 9s Cryptosporidium parvum isolates from humans and animals by a polymerase chain reaction/restriction fragment length polymorphism method based on the thrombospondin-related anonymous protein 2 gene sequence. Used as a molecular marker, this method can differentiate ...

  9. THE INFECTIVITY OF CRYPTOSPORIDIUM PARVUM IN HEALTHY VOLUNTEERS

    EPA Science Inventory

    Background. Small numbers of Cryptosporidium parvum oocysts can contaminate even treated drinking water, and ingestion of oocysts can cause diarrheal disease in normal as well as immunocompromised hosts. Since the number of organisms necessary to cause infection in humans is unkn...

  10. 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. PMID:26176919

  11. 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...

  12. A highly divergent 33 kDa Cryptosporidium parvum antigen

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Previous studies comparing the genome sequences of Cryptosporidium parvum with C. hominis identified a number of highly divergent genes that might reflect positive selection for host specificity. In the present study, a C. parvum sequence, namely cgd8-5370, whose amino acid sequence differs appreci...

  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. PMID:27084487

  14. Genetic richness and diversity in Cryptosporidium hominis and C. parvum reveals major knowledge gaps and a need for the application of "next generation" technologies--research review.

    PubMed

    Jex, Aaron R; Gasser, Robin B

    2010-01-01

    Cryptosporidium species (apicomplexan protists) are a major cause of diarrhoeal disease (= cryptosporidiosis) in humans worldwide. The impact of cryptosporidiosis is also compounded by the spread of HIV/AIDS and a lack of cost-effective anti-cryptosporidial chemotherapeutics or vaccines. Mitigation of the impact of cryptosporidiosis in humans needs to focus on prevention and control strategies, built on a sound understanding of the epidemiology of Cryptosporidium species. Refined epidemiological studies rely on the use of molecular tools employing informative genetic markers. Currently, the 60-kDa glycoprotein gene (gp60) is the most suitable and widely used genetic marker for Cryptosporidium species infecting humans. Here, we undertake an analysis of all publicly-available gp60 sequence data and associated literature for C. hominis and C. parvum, and yield useful insights into the richness, diversity and distribution of genetic variants, and link these variants to human cryptosporidiosis. This global analysis reveals that, despite high genetic richness in Cryptosporidium isolates from humans, there is a surprisingly low diversity. It also highlights limited knowledge about the genetics of cryptosporidiosis in developing nations and in many animals that might act as infection sources. Clearly, there is a major need for more comprehensive studies of Cryptosporidium infecting humans and other animals in Africa and Asia. As molecular technologies improve and become affordable, future studies should utilize "next generation" sequencing and bioinformatic platforms to conduct comparative 'genome sequence surveys' to test the validity of current genetic classifications based on gp60 data. Complemented by in vitro and in vivo investigations, these biotechnological advances will also assist significantly in the search for new intervention strategies against human cryptosporidiosis.

  15. 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.

  16. 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.

  17. Comparison of Cryptosporidium parvum and Cryptosporidium wrairi by reactivity with monoclonal antibodies and ability to infect severe combined immunodeficient mice.

    PubMed Central

    Chrisp, C E; Mason, P; Perryman, L E

    1995-01-01

    Twenty-three monoclonal antibodies raised to Cryptosporidium parvum and 12 raised to C. wrairi reacted with equal intensity with the heterologous species. Despite demonstration of a close immunologic relationship between these two species, C. wrairi did not induce persistent infection in severe combined immunodeficient mice as did C. parvum. PMID:7806379

  18. 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

  19. 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