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 Cryptosporidium concentrations as estimated by the meta-analysis, but when DWIs were influenced by agricultural runoff or wildlife, there was a poor relationship. Average recovery values were available for 6 out of 22 Cryptosporidium concentration data sets and concomitant analysis demonstrated no changes in trends, with and without correction. Nevertheless, recovery assays performed along with every oocyst count would have enhanced the precision of this work. Based on our findings, the use of annual averages of E. coli concentrations as a surrogate for Cryptosporidium concentrations can result in an inaccurate estimate of the Cryptosporidium risk for agriculture impacted drinking water intakes or for intakes with more distant wastewater sources. Studies of upstream fecal pollution sources are recommended for drinking water suppliers to improve their interpretation of source water quality data. Copyright © 2014 Elsevier Ltd. All rights reserved.

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

... 40 Protection of Environment 23 2014-07-01 2014-07-01 false Special notice for repeated failure to conduct monitoring of the source water for Cryptosporidium and for failure to determine bin classification or mean Cryptosporidium level. 141.211 Section 141.211 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS ...

[Investigation on contamination of Giardia and Cryptosporidium in drinking water in Jiangsu Province].

PubMed

Bi-Xian, N I; Ming-Xue, S; Xiang-Zhen, X U; Xiao-Ting, W; Yang, D; Xiao-Lin, J

2017-05-17

Objective To know the contamination status of Giardia lamblia and Cryptosporidium in drinking water of Jiangsu Province, so as to provide the evidence for producing hygiene and safety drinking water. Methods A total of 28 water plants of 13 cities in Jiangsu Province were selected, and the source water (10 L), chlorinated water (100 L) and tap water (100 L) were collected separately in each site. The water samples were then treated by filtration, washing, centrifuging concentration, immune magnetic separation, and immunofluorescent assay, to detect the numbers of Giardia cysts and Cryptosporidium oocysts. Results Totally 84 samples from 13 cities were collected, including 28 source water, 28 chlorinated water and 28 tap water samples. Among the chlorinated water and tap water samples, no Giardia cysts and Cryptosporidium oocysts were found. However, Giardia cysts were detected in 3 (10.71%, 3/28) source water samples (Yancheng, Lianyungang, Changzhou cities), with the density of 1 cyst/10 L of all. Cryptosporidium oocysts were also detected in 3 (10.71%, 3/28) source water samples (Nanjing, Zhenjiang, Yangzhou cities), with the density of 1 oocyst/10 L of all. Conclusions The source water in partial areas of Jiangsu Province has been contaminated by Giardia and Cryptosporidium . To ensure the safety of drinking, the regulation of source water and surveillance of drinking water should be strengthened.

Adaptive management for mitigating Cryptosporidium risk in source water: a case study in an agricultural catchment in South Australia.

PubMed

Bryan, Brett A; Kandulu, John; Deere, Daniel A; White, Monique; Frizenschaf, Jacqueline; Crossman, Neville D

2009-07-01

Water-borne pathogens such as Cryptosporidium pose a significant human health risk and catchments provide the first critical pollution 'barrier' in mitigating risk in drinking water supply. In this paper we apply an adaptive management framework to mitigating Cryptosporidium risk in source water using a case study of the Myponga catchment in South Australia. Firstly, we evaluated the effectiveness of past water quality management programs in relation to the adoption of practices by landholders using a socio-economic survey of land use and management in the catchment. The impact of past management on the mitigation of Cryptosporidium risk in source water was also evaluated based on analysis of water quality monitoring data. Quantitative risk assessment was used in planning the next round of management in the adaptive cycle. Specifically, a pathogen budget model was used to identify the major remaining sources of Cryptosporidium in the catchment and estimate the mitigation impact of 30 alternative catchment management scenarios. Survey results show that earlier programs have resulted in the comprehensive adoption of best management practices by dairy farmers including exclusion of stock from watercourses and effluent management from 2000 to 2007. Whilst median Cryptosporidium concentrations in source water have decreased since 2004 they remain above target levels and put pressure on other barriers to mitigate risk, particularly the treatment plant. Non-dairy calves were identified as the major remaining source of Cryptosporidium in the Myponga catchment. The restriction of watercourse access of non-dairy calves could achieve a further reduction in Cryptosporidium export to the Myponga reservoir of around 90% from current levels. The adaptive management framework applied in this study was useful in guiding learning from past management, and in analysing, planning and refocusing the next round of catchment management strategies to achieve water quality targets.

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

... classification or mean Cryptosporidium level must contain the following language: We are required to monitor the... or mean Cryptosporidium level. 141.211 Section 141.211 Protection of Environment ENVIRONMENTAL... Cryptosporidium level. (a) When is the special notice for repeated failure to monitor to be given? The owner or...

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.

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.

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.

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.g., muskrat I and II genotype) do not pose significant infection risks to humans. Consequently, from a human health perspective, land use practices in agricultural watersheds that create opportunities for wildlife to flourish should not be rejected solely on the basis of their potential to increase relative proportions of wildlife fecal contamination in surface water. The present study suggests that mitigating livestock fecal pollution in surface water in this region would likely reduce human infection risks associated with Cryptosporidium and other zoonotic pathogens. PMID:23124241

[Investigation on contamination of Cryptosporidium and Giardia in drinking water and environmental water in Shanghai].

PubMed

Zhang, Xiao-Ping; He, Yan-Yan; Zhu, Qian; Ma, Xiao-Jiang; Cai, Li

2010-12-30

To understand the contamination status of Cryptosporidium sp. and Giardia lamblia in drinking water, source water and environmental water in Shanghai. All water samples collected from drinking water, source water and environmental water were detected by a procedure of micromembrane filtration, immune magnetic separation (IMS), and immunofluorescent assay (IFA). Cryptosporidium oocysts and Giardia cysts were not found in 156 samples of the drinking water including finished water, tap water, or pipe water for directly drinking in communities. Among 70 samples either source water of water plants (15 samples), environmental water from Huangpu River(25), canal water around animal sheds(15), exit water from waste-water treatment plants(9), or waste water due to daily life(6), Cryptosporidium oocysts were detected in 1(6.7%), 2(8.0%), 7(46.7%), 1(11.1%), and 1(16.7%) samples, respectively; and Giardia cysts were detected in 1(6.7%), 3(12.0%), 6 (40.0%), 2(22.2%), and 2(33.3%), respectively. The positive rate of Cryptosporidium oocysts and Giardia cysts was 17.1% (12/70) and 20.0% (14/70), respectively. No Cryptosporidium oocysts and Giardia cysts have been detected in drinking water, but found in source water and environmental water samples in Shanghai.

Epidemiology of equine Cryptosporidium and Giardia infections.

PubMed

Xiao, L; Herd, R P

1994-01-01

Prevalence and infection patterns of Cryptosporidium and Giardia infections in horses were studied by a direct immunofluorescence staining method. Faecal examinations of 222 horses of different age groups revealed Cryptosporidium infection rates of 15-31% in 66 foals surveyed in central Ohio, southern Ohio and central Kentucky, USA. Only 1 of 39 weanlings, 0 of 46 yearlings, and 0 of 71 mares were positive. Giardia infection was found in all age groups, although the infection rates for foals were higher (17-35%). Chronological study of infection in 35 foals showed that foals started to excrete Cryptosporidium oocysts between 4 and 19 weeks and Giardia cysts between 2 and 22 weeks of age. The cumulative infection rates of Cryptosporidium and Giardia in foals were each 71%. Some foals were concurrently infected with both parasites and excretion of oocysts or cysts was intermittent and long-lasting. The longest duration of excretion was 14 weeks for Cryptosporidium and 16 weeks for Giardia. Excretion of Cryptosporidium oocysts stopped before weaning, while excretion of Giardia cysts continued thereafter. Infected foals were considered the major source of Cryptosporidium infection in foals, whereas infected mares were deemed the major source of Giardia infection in foals. The high infection rate of Giardia in nursing mares suggested a periparturient relaxation of immunity. The results indicated that Cryptosporidium and Giardia infections are common in horses.

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

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...

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...

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...

40 CFR 141.716 - Source toolbox components.

Code of Federal Regulations, 2011 CFR

2011-07-01

... undertake to reduce source water Cryptosporidium levels. The plan must explain how the actions are expected... Section 141.716 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Requirements...

40 CFR 141.716 - Source toolbox components.

Code of Federal Regulations, 2010 CFR

2010-07-01

... undertake to reduce source water Cryptosporidium levels. The plan must explain how the actions are expected... Section 141.716 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Requirements...

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.

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...

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...

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

PubMed Central

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

2013-01-01

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

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

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, Thierry M.; Aguirre, A.A.; Ellis, D.M.; Murakawa, Shawn K. K.; Morris, Robert

1997-01-01

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

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.

Prevalence and molecular characterization of bovine Cryptosporidium from dairy cows in Northern Thailand.

PubMed

Inpankaew, Tawin; Jiyipong, Tawisa; Sunanta, Chainirun; Kengradomkij, Chanya; Pinyopanuwat, Nongnuch; Jittapalapong, Sathaporn

2017-12-20

Cryptosporidiosis is a common protozoan infection in humans and domestic animals. It is the culprit for significant neonatal morbidity in cattle as well as weight loss and delayed growth, which leads to large economic losses in the farming industry. Furthermore, bovine Cryptosporidium is also a principal source of human Cryptosporidium infections. The purpose of this study is to determine prevalence and genotype of Cryptosporidium spp. from feces of dairy cows from the northern parts of Thailand (Chiang Rai, Chiang Mai, and Lumpang provinces). A total of 500 fecal samples were collected directly from the rectum and they were examined for potential presence of Cryptosporidium infection by using tests such as DMSO-modified acid fast stain, immunofluorescence antibody test (IFAT) and polymerase chain reaction (PCR). The prevalence of Cryptosporidium spp. was 5% by DMSO-modified acid fast stain, 7% by IFAT and 7.6% by PCR respectively. The main genotypes of Cryptosporidium spp. identified were Cryptosporidium parvum and Cryptosporidium bovis. Therefore, as a result of this study, it can be said that, due to the potential cross-species transmission of Cryptosporidium parvum, infected dairy cows may pose a potential zoonotic risk to humans.

Environmental Transport of Emerging Human-Pathogenic Cryptosporidium Species and Subtypes through Combined Sewer Overflow and Wastewater

PubMed Central

Huang, Chengchen; Hu, Yue; Wang, Lin; Wang, Yuanfei; Li, Na; Guo, Yaqiong; Xiao, Lihua

2017-01-01

ABSTRACT The environmental transport of Cryptosporidium spp. through combined sewer overflow (CSO) and the occurrence of several emerging human-pathogenic Cryptosporidium species in developing countries remain unclear. In this study, we collected 40 CSO samples and 40 raw wastewater samples from Shanghai, China, and examined them by PCR and DNA sequencing for Cryptosporidium species (targeting the small subunit rRNA gene) and Giardia duodenalis (targeting the triosephosphate isomerase, β-giardin, and glutamate dehydrogenase genes) and Enterocytozoon bieneusi (targeting the ribosomal internal transcribed spacer) genotypes. Human-pathogenic Cryptosporidium species were further subtyped by sequence analysis of the 60-kDa glycoprotein gene, with additional multilocus sequence typing on the emerging zoonotic pathogen Cryptosporidium ubiquitum. Cryptosporidium spp., G. duodenalis, and E. bieneusi were detected in 12 and 15, 33 and 32, and 37 and 40 CSO and wastewater samples, respectively, including 10 Cryptosporidium species, 3 G. duodenalis assemblages, and 8 E. bieneusi genotypes. In addition to Cryptosporidium hominis and Cryptosporidium parvum, two new pathogens identified in industrialized nations, C. ubiquitum and Cryptosporidium viatorum, were frequently detected. The two novel C. ubiquitum subtype families identified appeared to be genetic recombinants of known subtype families. Similarly, the dominant group 1 E. bieneusi genotypes and G. duodenalis subassemblage AII are known human pathogens. The similar distribution of human-pathogenic Cryptosporidium species and E. bieneusi and G. duodenalis genotypes between wastewater and CSO samples reaffirms that storm overflow is potentially a significant contamination source of pathogens in surface water. The frequent identification of C. ubiquitum and C. viatorum in urban wastewater suggests that these newly identified human pathogens may be endemic in China. IMPORTANCE Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi are major waterborne pathogens. Their transport into surface water through combined sewer overflow, which remains largely untreated in developing countries, has not been examined. In addition, the identification of these pathogens to genotypes and subtypes in urban storm overflow and wastewater is necessary for rapid and accurate assessment of pathogen transmission in humans and transport in the environment. Data from this study suggest that, like untreated urban wastewater, combined sewer overflow is commonly contaminated with human-pathogenic Cryptosporidium, G. duodenalis, and E. bieneusi genotypes and subtypes, and urban storm overflow potentially plays a significant role in the contamination of drinking source water and recreational water with human pathogens. They also indicate that Cryptosporidium ubiquitum and Cryptosporidium viatorum, two newly identified human pathogens, may be common in China, and genetic recombination can lead to the emergence of novel C. ubiquitum subtype families. PMID:28600310

Estimating Cryptosporidium and Giardia disease burdens for children drinking untreated groundwater in a rural population in India.

PubMed

Daniels, Miles E; Smith, Woutrina A; Jenkins, Marion W

2018-01-01

In many low-income settings, despite improvements in sanitation and hygiene, groundwater sources used for drinking may be contaminated with enteric pathogens such as Cryptosporidium and Giardia, which remain important causes of childhood morbidity. In this study, we examined the contribution of diarrhea caused by Cryptosporidium and Giardia found in groundwater sources used for drinking to the total burden of diarrheal disease among children < 5 in rural India. We studied a population of 3,385 children < 5 years of age in 100 communities of Puri District, Odisha, India. We developed a coupled quantitative microbial risk assessment (QMRA) and susceptible-infected-recovered (SIR) population model based on observed levels of Cryptosporidium and Giardia in improved groundwater sources used for drinking and compared the QMRA-SIR estimates with independently measured all-cause (i.e., all fecal-oral enteric pathogens and exposure pathways) child diarrhea prevalence rates observed in the study population during two monsoon seasons (2012 and 2013). We used site specific and regional studies to inform assumptions about the human pathogenicity of the Cryptosporidium and Giardia species present in local groundwater. In all three human pathogenicity scenarios evaluated, the mean daily risk of Cryptosporidium or Giardia infection (0.06-1.53%), far exceeded the tolerable daily risk of infection from drinking water in the US (< 0.0001%). Depending on which protozoa species were present, median estimates of daily child diarrhea prevalence due to either Cryptosporidium or Giardia infection from drinking water was as high as 6.5% or as low as < 1% and accounted for at least 2.9% and as much as 65.8% of the all-cause diarrhea disease burden measured in children < 5 during the study period. Cryptosporidium tended to account for a greater share of estimated waterborne protozoa infections causing diarrhea than did Giardia. Diarrhea prevalence estimates for waterborne Cryptosporidium infection appeared to be most sensitive to assumptions about the probability of infection from ingesting a single parasite (i.e. the rate parameter in dose-response model), while Giardia infection was most sensitive to assumptions about the viability of parasites detected in groundwater samples. Protozoa in groundwater drinking sources in rural India, even at low concentrations, especially for Cryptosporidium, may account for a significant portion of child diarrhea morbidity in settings were tubewells are used for drinking water and should be more systematically monitored. Preventing diarrheal disease burdens in Puri District and similar settings will benefit from ensuring water is microbiologically safe for consumption and consistent and effective household water treatment is practiced.

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 highly at risk to Cryptosporidium/Giardia infections. Copyright © 2016 Elsevier B.V. All rights reserved.

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.

Estimating Cryptosporidium and Giardia disease burdens for children drinking untreated groundwater in a rural population in India

PubMed Central

2018-01-01

Background In many low-income settings, despite improvements in sanitation and hygiene, groundwater sources used for drinking may be contaminated with enteric pathogens such as Cryptosporidium and Giardia, which remain important causes of childhood morbidity. In this study, we examined the contribution of diarrhea caused by Cryptosporidium and Giardia found in groundwater sources used for drinking to the total burden of diarrheal disease among children < 5 in rural India. Methodology/Principal findings We studied a population of 3,385 children < 5 years of age in 100 communities of Puri District, Odisha, India. We developed a coupled quantitative microbial risk assessment (QMRA) and susceptible-infected-recovered (SIR) population model based on observed levels of Cryptosporidium and Giardia in improved groundwater sources used for drinking and compared the QMRA-SIR estimates with independently measured all-cause (i.e., all fecal-oral enteric pathogens and exposure pathways) child diarrhea prevalence rates observed in the study population during two monsoon seasons (2012 and 2013). We used site specific and regional studies to inform assumptions about the human pathogenicity of the Cryptosporidium and Giardia species present in local groundwater. In all three human pathogenicity scenarios evaluated, the mean daily risk of Cryptosporidium or Giardia infection (0.06–1.53%), far exceeded the tolerable daily risk of infection from drinking water in the US (< 0.0001%). Depending on which protozoa species were present, median estimates of daily child diarrhea prevalence due to either Cryptosporidium or Giardia infection from drinking water was as high as 6.5% or as low as < 1% and accounted for at least 2.9% and as much as 65.8% of the all-cause diarrhea disease burden measured in children < 5 during the study period. Cryptosporidium tended to account for a greater share of estimated waterborne protozoa infections causing diarrhea than did Giardia. Diarrhea prevalence estimates for waterborne Cryptosporidium infection appeared to be most sensitive to assumptions about the probability of infection from ingesting a single parasite (i.e. the rate parameter in dose-response model), while Giardia infection was most sensitive to assumptions about the viability of parasites detected in groundwater samples. Conclusions/Significance Protozoa in groundwater drinking sources in rural India, even at low concentrations, especially for Cryptosporidium, may account for a significant portion of child diarrhea morbidity in settings were tubewells are used for drinking water and should be more systematically monitored. Preventing diarrheal disease burdens in Puri District and similar settings will benefit from ensuring water is microbiologically safe for consumption and consistent and effective household water treatment is practiced. PMID:29377884

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. Copyright © 2015 Elsevier Ltd. All rights reserved.

Occurrence of Cryptosporidium and Giardia genotypes and subtypes in raw and treated water in Portugal.

PubMed

Lobo, M L; Xiao, L; Antunes, F; Matos, O

2009-06-01

Waterborne outbreaks of diarrhoeal illness reported worldwide are mostly associated with Cryptosporidium spp. and Giardia spp. Their presence in aquatic systems makes it essential to develop preventive strategies for water and food safety. This study was undertaken to monitor the presence of Cryptosporidium and Giardia in a total of 175 water samples, including raw and treated water from both surface and ground sources in Portugal. The samples were processed according to USEPA Method 1623 for immunomagnetic separation (IMS) of Cryptosporidium oocysts and Giardia cysts, followed by detection of oocysts/cysts by immunofluorecence (IFA) microscopy, PCR-based techniques were done on all water samples collected. Out of 175 samples, 81 (46.3%) were positive for Cryptosporidium and 67 (38.3%) for Giardia by IFA. Cryptosporidium spp. and G. duodenalis genotypes were identified by PCR in 37 (21.7%) and 9 (5.1%) water samples, respectively. C. parvum was the most common species (78.9%), followed by C. hominis (13.2%), C. andersoni (5.3%), and C. muris (2.6%). Subtype IdA15 was identified in all C. hominis-positive water samples. Subtyping revealed the presence of C. parvum subtypes IIaA15G2R1, IIaA16G2R1 and IIdA17G1. Giardia duodenalis subtype A1 was identified. The results of the present study suggest that Cryptosporidium spp. and Giardia spp. were widely distributed in source water and treated water in Portugal. Moreover, the results obtained indicate a high occurrence of human-pathogenic Cryptosporidium genotypes and subtypes in raw and treated water samples. Thus, water can be a potential vehicle in the transmission of cryptosporidiosis, and giardiasis of humans and animals in Portugal.

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 areas was not found to be a significant contributor of Cryptosporidium, however, it was found to be a Giardia source.

Molecular characterization of Cryptosporidium from animal sources in Qinghai province of China.

PubMed

Karanis, Panagiotis; Plutzer, Judit; Halim, Norhaliza Abdul; Igori, Khatanbaatar; Nagasawa, Hideyuki; Ongerth, Jerry; Liqing, Ma

2007-11-01

The presence of Cryptosporidium oocysts in 20 zoo animals of the Xining Zoo, 16 farm yaks and 42 farm goats in Qinghai province, China was investigated by an immunofluorescence test (IFT). The species and/or genotypes were determined by nested polymerase chain reaction (PCR) and sequence analysis of a fragment of the small subunit (SSU) rRNA gene. Cryptosporidium oocysts were found in 16 zoo animals, 2 yaks, and 15 goats by IFT. The IFT positive samples were further investigated by PCR, and 16 of them were found to be positive by that method also. Sequence analysis of the PCR products derived from Cryptosporidium oocysts from Black leopard (Panthera pardus), Heijing He (Grus nigricollis), Barbary sheep (Ammotragus lervia), Takin (Budorcas taxicolor), Lesser panda (Ailurus fulgens), and White-eared pheasant (Crossoptilon crossoptilon) fecal samples matched that of Cryptosporidium parvum mouse genotype. Sequence analyses of other PCR products were consistent with cervine genotype Cryptosporidium from Ibex (Capra ibex), a novel Cryptosporidium genotype from a wild yak (Bos mutus), C. bovis-like genotype from one goat sample and also a novel Cryptosporidium genotype from one other separate goat sample. The present work reports the first data on Cryptosporidium infections in animals from the Qinghai province of mountainous central western China and the first findings of the 'cervine' genotype in Capra ibex, C. bovis-like genotype and the new Cryptosporidium spp. in farm goat and in wild yak.

Captive-bred neotropical birds diagnosed with Cryptosporidium Avian genotype III.

PubMed

Silva Novaes, Ricardo; Pires, Marcus Sandes; Sudré, Adriana Pittella; Bergamo do Bomfim, Teresa Cristina

2018-02-01

Currently, there are only three valid species of Cryptosporidium infecting avian hosts, namely, Cryptosporidium meleagridis, Cryptosporidium baileyi, Cryptosporidium galli and Cryptosporidium avium in addition to 12 genotypes of unknown species status. The objectives of this study were to microscopically diagnose the presence of Cryptosporidium in birds from a commercial aviary located in Rio de Janeiro, Brazil; genotypically characterize species and/or genotypes of genus Cryptosporidum; and conduct sequencing and phylogenetic analyses to compare the obtained DNA sequences with those deposited in GenBank. A total of 85 fecal samples were collected from wild captive-bred birds: 48 of family Psittacidae and 37 of family Ramphastidae. Initially, a search for the presence of Cryptosporidium sp. oocysts was conducted using the centrifugal-flotation in saturated sugar solution technique, after that, the collected samples were analyzed microscopically. Cryptosporidium infections were only detected in 24.32% of samples belonging to the family Ramphastidae. DNA was extracted from positive samples and molecular diagnostics was applied targeting the 18S rRNA gene, followed by sequencing and phylogenetic analysis. The Cryptosporidium Avian genotype III was diagnosed in this study more closely related to the gastric species. This is the first record of Cryptosporidium Avian genotype III in order Piciformes and family Ramphastidae, where three host species (Ramphastus toco, Ramphastus tucanus, and Pteroglossus bailloni) were positive for the etiologic agent. Based on the molecular data obtained, these wild birds raised in captivity do not represent a source of human cryptosporidiosis, considering that Cryptosporidium Avian genotype III does not constitute a zoonosis. Copyright © 2017. Published by Elsevier B.V.

Molecular Surveillance of Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi by Genotyping and Subtyping Parasites in Wastewater

PubMed Central

Li, Na; Xiao, Lihua; Wang, Lin; Zhao, Shuming; Zhao, Xukun; Duan, Liping; Guo, Meijin; Liu, Lili; Feng, Yaoyu

2012-01-01

Background Despite their wide occurrence, cryptosporidiosis and giardiasis are considered neglected diseases by the World Health Organization. The epidemiology of these diseases and microsporidiosis in humans in developing countries is poorly understood. The high concentration of pathogens in raw sewage makes the characterization of the transmission of these pathogens simple through the genotype and subtype analysis of a small number of samples. Methodology/Principal Findings The distribution of genotypes and subtypes of Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi in 386 samples of combined sewer systems from Shanghai, Nanjing and Wuhan and the sewer system in Qingdao in China was determined using PCR-sequencing tools. Eimeria spp. were also genotyped to assess the contribution of domestic animals to Cryptosporidium spp., G. duodenalis, and E. bieneusi in wastewater. The high occurrence of Cryptosporidium spp. (56.2%), G. duodenalis (82.6%), E. bieneusi (87.6%), and Eimeria/Cyclospora (80.3%) made the source attribution possible. As expected, several human-pathogenic species/genotypes, including Cryptosporidium hominis, Cryptosporidium meleagridis, G. duodenalis sub-assemblage A-II, and E. bieneusi genotype D, were the dominant parasites in wastewater. In addition to humans, the common presence of Cryptosporidium spp. and Eimeria spp. from rodents indicated that rodents might have contributed to the occurrence of E. bieneusi genotype D in samples. Likewise, the finding of Eimeria spp. and Cryptosporidium baileyi from birds indicated that C. meleagridis might be of both human and bird origins. Conclusions/Significance The distribution of Cryptosporidium species, G. duodenalis genotypes and subtypes, and E. bieneusi genotypes in urban wastewater indicates that anthroponotic transmission appeared to be important in epidemiology of cryptosporidiosis, giardiasis, and microsporidiosis in the study areas. The finding of different distributions of subtypes between Shanghai and Wuhan was indicative of possible differences in the source of C. hominis among different areas in China. PMID:22970334

Understanding human infectious Cryptosporidium risk in drinking water supply catchments.

PubMed

Swaffer, Brooke; Abbott, Hayley; King, Brendon; van der Linden, Leon; Monis, Paul

2018-07-01

Treating drinking water appropriately depends, in part, on the robustness of source water quality risk assessments, however quantifying the proportion of infectious, human pathogenic Cryptosporidium oocysts remains a significant challenge. We analysed 962 source water samples across nine locations to profile the occurrence, rate and timing of infectious, human pathogenic Cryptosporidium in surface waters entering drinking water reservoirs during rainfall-runoff conditions. At the catchment level, average infectivity over the four-year study period reached 18%; however, most locations averaged <5%. The maximum recorded infectivity fraction within a single rainfall runoff event was 65.4%, and was dominated by C. parvum. Twenty-two Cryptosporidium species and genotypes were identified using PCR-based molecular techniques; the most common being C. parvum, detected in 23% of water samples. Associations between landuse and livestock stocking characteristics with Cryptosporidium were determined using a linear mixed-effects model. The concentration of pathogens in water were significantly influenced by flow and dominance of land-use by commercial grazing properties (as opposed to lifestyle properties) in the catchment (p < 0.01). Inclusion of measured infectivity and human pathogenicity data into a quantitative microbial risk assessment (QMRA) could reduce the source water treatment requirements by up to 2.67 log removal values, depending on the catchment, and demonstrated the potential benefit of collating such data for QMRAs. Copyright © 2018 Elsevier Ltd. All rights reserved.

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...

A Randomized Controlled Trial to Assess the Impact of Ceramic Water Filters on Prevention of Diarrhea and Cryptosporidiosis in Infants and Young Children-Western Kenya, 2013.

PubMed

Morris, Jamae Fontain; Murphy, Jennifer; Fagerli, Kirsten; Schneeberger, Chandra; Jaron, Peter; Moke, Fenny; Juma, Jane; Ochieng, J Ben; Omore, Richard; Roellig, Dawn; Xiao, Lihua; Priest, Jeffrey W; Narayanan, Jothikumar; Montgomery, Joel; Hill, Vince; Mintz, Eric; Ayers, Tracy L; O'Reilly, Ciara E

2018-04-02

Cryptosporidium is a leading cause of diarrhea among Kenyan infants. Ceramic water filters (CWFs) are used for household water treatment. We assessed the impact of CWFs on diarrhea, cryptosporidiosis prevention, and water quality in rural western Kenya. A randomized, controlled intervention trial was conducted in 240 households with infants 4-10 months old. Twenty-six weekly household surveys assessed infant diarrhea and health facility visits. Stool specimens from infants with diarrhea were examined for Cryptosporidium . Source water, filtered water, and filter retentate were tested for Cryptosporidium and/or microbial indicators. To estimate the effect of CWFs on health outcomes, logistic regression models using generalized estimating equations were performed; odds ratios (ORs) and 95% confidence intervals (CIs) are reported. Households reported using surface water (36%), public taps (29%), or rainwater (17%) as their primary drinking water sources, with no differences in treatment groups. Intervention households reported less diarrhea (7.6% versus 8.9%; OR: 0.86 [0.64-1.16]) and significantly fewer health facility visits for diarrhea (1.0% versus 1.9%; OR: 0.50 [0.30-0.83]). In total, 15% of intervention and 12% of control stools yielded Cryptosporidium ( P = 0.26). Escherichia coli was detected in 93% of source water samples; 71% of filtered water samples met World Health Organization recommendations of < 1 E. coli /100 mL. Cryptosporidium was not detected in source water and was detected in just 2% of filter rinses following passage of large volumes of source water. Water quality was improved among CWF users; however, the short study duration and small sample size limited our ability to observe reductions in cryptosporidiosis.

Presence of Pathogens and Indicator Microbes at a Non-Point Source Subtropical Recreational Marine Beach ▿ †

PubMed Central

Abdelzaher, Amir M.; Wright, Mary E.; Ortega, Cristina; Solo-Gabriele, Helena M.; Miller, Gary; Elmir, Samir; Newman, Xihui; Shih, Peter; Bonilla, J. Alfredo; Bonilla, Tonya D.; Palmer, Carol J.; Scott, Troy; Lukasik, Jerzy; Harwood, Valerie J.; McQuaig, Shannon; Sinigalliano, Chris; Gidley, Maribeth; Plano, Lisa R. W.; Zhu, Xiaofang; Wang, John D.; Fleming, Lora E.

2010-01-01

Swimming in ocean water, including ocean water at beaches not impacted by known point sources of pollution, is an increasing health concern. This study was an initial evaluation of the presence of indicator microbes and pathogens and the association among the indicator microbes, pathogens, and environmental conditions at a subtropical, recreational marine beach in south Florida impacted by non-point sources of pollution. Twelve water and eight sand samples were collected during four sampling events at high or low tide under elevated or reduced solar insolation conditions. The analyses performed included analyses of fecal indicator bacteria (FIB) (fecal coliforms, Escherichia coli, enterococci, and Clostridium perfringens), human-associated microbial source tracking (MST) markers (human polyomaviruses [HPyVs] and Enterococcus faecium esp gene), and pathogens (Vibrio vulnificus, Staphylococcus aureus, enterovirus, norovirus, hepatitis A virus, Cryptosporidium spp., and Giardia spp.). The enterococcus concentrations in water and sand determined by quantitative PCR were greater than the concentrations determined by membrane filtration measurement. The FIB concentrations in water were below the recreational water quality standards for three of the four sampling events, when pathogens and MST markers were also generally undetectable. The FIB levels exceeded regulatory guidelines during one event, and this was accompanied by detection of HPyVs and pathogens, including detection of the autochthonous bacterium V. vulnificus in sand and water, detection of the allochthonous protozoans Giardia spp. in water, and detection of Cryptosporidium spp. in sand samples. The elevated microbial levels were detected at high tide and under low-solar-insolation conditions. Additional sampling should be conducted to further explore the relationships between tidal and solar insolation conditions and between indicator microbes and pathogens in subtropical recreational marine waters impacted by non-point source pollution. PMID:19966020

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...

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...

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...

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...

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

Integrated cryptosporidium assay to determine oocyst density, infectivity, and genotype for risk assessment of source and reuse water.

PubMed

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

2015-05-15

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. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

How clean must our drinking water be: the importance of protective immunity.

PubMed

Frost, Floyd J; Roberts, Melissa; Kunde, Twila R; Craun, Gunther; Tollestrup, Kristine; Harter, Lucy; Muller, Tim

2005-03-01

Cryptosporidium parvum is an important cause of epidemic diarrhea. Few studies have assessed whether serological evidence of prior infection in adults is related to a reduced occurrence of enteric illness. Serum samples and enteric illness event data were obtained in 2000 and 2001 from 326 people served by 1 of 2 unfiltered surface sources or 1 groundwater source. In 2001, filtration was initiated at 1 of the surface sources. Poisson regression related illness episodes with serological responses to the 15/17- and 27-kDa Cryptosporidium antigen groups. Subjects with moderately strong responses to the 15/17-kDa antigen had <65% of the risk of all 1-3-day episodes of diarrheal or gastrointestinal illness and <40% of the risk of all >/=4-day episodes, compared with subjects without a moderately strong response. Water source, change in water treatment, and very weak responses were unrelated to illness events. Endemic Cryptosporidium infections are a common cause of diarrheal and gastrointestinal illness in persons without a moderately strong response to the 15/17-kDa antigen group. Users of surface-derived drinking water are more likely to have strong serological responses to this antigen group and may be at a lower risk of endemic gastrointestinal illness caused by Cryptosporidium infection.

Biological and genetic characterization of Cryptosporidium spp. and Giardia duodenalis isolates from five hydrographical basins in northern Portugal.

PubMed

Almeida, André; Moreira, Maria João; Soares, Sónia; de Lurdes Delgado, Maria; Figueiredo, João; Magalhães, Elisabete Silva; Castro, António; Viana Da Costa, Alexandra; Correia da Costa, José Manuel

2010-06-01

To understand the situation of water contamination with Cryptosporidium spp. and Giardia spp. in the northern region of Portugal, we have established a long-term program aimed at pinpointing the sources of surface water and environmental contamination, working with the water-supply industry. Here, we describe the results obtained with raw water samples collected in rivers of the 5 hydrographical basins. A total of 283 samples were analyzed using the Method 1623 EPA, USA. Genetic characterization was performed by PCR and sequencing of genes 18S rRNA of Cryptosporidium spp. and beta-giardin of Giardia spp. Infectious stages of the protozoa were detected in 72.8% (206 of 283) of the water samples, with 15.2% (43 of 283) positive for Giardia duodenalis cysts, 9.5% (27 of 283) positive for Cryptosporidium spp. oocysts, and 48.1% (136 of 283) samples positive for both parasites. The most common zoonotic species found were G. duodenalis assemblages A-I, A-II, B, and E genotypes, and Cryptosporidium parvum, Cryptosporidium andersoni, Cryptosporidium hominis, and Cryptosporidium muris. These results suggest that cryptosporidiosis and giardiasis are important public health issues in northern Portugal. To the authors' knowledge, this is the first report evaluating the concentration of environmental stages of Cryptosporidium and Giardia in raw water samples in the northern region of Portugal.

Comparison of human and southern sea otter (Enhydra lutris nereis) health risks for infection with protozoa in nearshore waters.

PubMed

Adell, A D; McBride, G; Wuertz, S; Conrad, P A; Smith, W A

2016-11-01

Cryptosporidium and Giardia spp. are waterborne, fecally-transmitted pathogens that cause economic loss due to gastroenteritis and beach closures. We applied quantitative microbial risk assessment (QMRA) to determine the health risks for humans and sea otters due to waterborne exposure of Cryptosporidium and Giardia spp. when swimming in three types of surface waters: river, stormwater and wastewater effluent during the wet and dry seasons in the central coast of California. This is the first application of QMRA to estimate both the probability of infection in Southern sea otters and the probability of illness in humans, using microbial source tracking (MST) as a variable. Children swimming close to stormwater discharges had an estimated Cryptosporidium-associated illness probability that exceeded the accepted U.S. EPA criteria (32 illnesses/1000 swimmers or 3.2%). Based on the assumption that sea otters are as susceptible as humans to Cryptosporidium infection, the infection probabilities were close to 2% and 16% when sea otters were swimming at the end of points of rivers and stormwater discharges, respectively. In the case of Giardia, infection probabilities of 11% and 23% were estimated for sea otters swimming at the end of point of wastewater discharges, assuming that sea otters are as susceptible as gerbils and humans, respectively. The results of this QMRA suggest that 1) humans and sea otters are at risk when swimming at outflow sites for rivers, stormwater and treated wastewater effluent; 2) reduced loads of viable protozoan cysts and oocysts in recreational water can lessen the probability of infection of humans and sea otters; and 3) the risk of infection of humans and sea otters can be reduced with the treatment of wastewater to decrease oocyst and cyst viability before effluent is released into the sea. Copyright © 2016 Elsevier Ltd. All rights reserved.

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

PubMed

Koompapong, Khuanchai; Sukthana, Yaowalark

2012-07-01

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

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

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. Copyright © 2015 Elsevier Inc. All rights reserved.

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

Cryptosporidium parvum infection involving novel genotypes in wildlife from lower New York State.

PubMed

Perz, J F; Le Blancq, S M

2001-03-01

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.

Participatory probabilistic assessment of the risk to human health associated with cryptosporidiosis from urban dairying in Dagoretti, Nairobi, Kenya.

PubMed

Grace, Delia; Monda, Joseph; Karanja, Nancy; Randolph, Thomas F; Kang'ethe, Erastus K

2012-09-01

We carried out a participatory risk assessment to estimate the risk (negative consequences and their likelihood) from zoonotic Cryptosporidium originating in dairy farms in urban Dagoretti, Nairobi to dairy farm households and their neighbours. We selected 20 households at high risk for Cryptosporidium from a larger sample of 300 dairy households in Dagoretti based on risk factors present. We then conducted a participatory mapping of the flow of the hazard from its origin (cattle) to human potential victims. This showed three main exposure pathways (food and water borne, occupational and recreational). This was used to develop a fault tree model which we parameterised using information from the study and literature. A stochastic simulation was used to estimate the probability of exposure to zoonotic cryptosporidiosis originating from urban dairying. Around 6 % of environmental samples were positive for Cryptosporidium. Probability of exposure to Cryptosporidium from dairy cattle ranged from 0.0055 for people with clinical acquired immunodeficiency syndrome in non-dairy households to 0.0102 for children under 5 years from dairy households. Most of the estimated health burden was born by children. Although dairy cattle are the source of Cryptosporidium, the model suggests consumption of vegetables is a greater source of risk than consumption of milk. In conclusion, by combining participatory methods with quantitative microbial risk assessment, we were able to rapidly, and with appropriate 'imprecision', investigate health risk to communities from Cryptosporidium and identify the most vulnerable groups and the most risky practices.

Microbiological Detection Systems for Molecular Analysis of Environmental Water and Soil Samples

EPA Science Inventory

Multiple detection systems are being targeted to track various species and genotypes of pathogens found in environmental samples with the overreaching goal of developing analytical separation and detection techniques for Salmonella enterica Serovars Typhi, Cryptosporidium parvum,...

EVALUATION OF SPIKING PROCEDURES FOR RECOVERY OF CRYPTOSPORIDIUM IN STREAM WATERS USING USEPA METHOD 1623

EPA Science Inventory

U.S. Environmental Protection Agency Method 1623 is widely used to monitor source waters and drinking water supplies for Cryptosporidium oocysts. Analyzing matrix spikes is an important component of Method 1623. Matrix spikes are used to determine the effect of the environmental...

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

USDA-ARS?s Scientific Manuscript database

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...

[Investigation of the presence of Cryptosporidium spp. in different water sources in Mersin province, Turkey].

PubMed

Aslan, Gönül; Bayram, Gül; Otağ, Feza; Direkel, Sahin; Taylan Özkan, Ayşegül; Ceber, Kemal; Emekdaş, Gürol

2012-01-01

Cryptosporidium is an intracellular protozoon that causes enteritis in human and animals. Contaminated water and food are the major sources for the transmission of oocysts via oral-fecal route. It is reported that the prevalence of cryptosporidiosis is higher in developing countries than developed countries because of inefficient sanitation and disinfection facilities for drinking water. The most frequently detected species is Cryptosporidium parvum leading to high morbidity in healthy subjects and also fatal infections in immunocompromised patients. The acid-fast staining method is widely used in the diagnosis of cryptosporidiosis. Nowadays, Cryptosporidium could easily be detected in water supplies and asymptomatic carriers by molecular techniques to obtain epidemiological data. In this study it was aimed to detect and identify Cryptosporidium oocysts in different water sources in Mersin province, Turkey. A total of 135 water samples (70 taps, 50 wells and 15 sewage) collected from city center (n= 25) and from Tarsus (n= 32), Mezitli (n= 33) and Karaduvar (n= 45) counties between March 2007 and May 2009 were included in the study. Water samples in 10 liter volumes, were filtered by 0.45 µm pore-sized membrane filter vacuum/ pressure pumping technique. Cryptosporidium oocysts in filtrates were detected by modified cold Kinyoun acid-fast stain (MCK) technique and also identified and typed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. MCK yielded three and PCR yielded seven positive results. All the strains were identified as C.parvum by PCR-RFLP method. All of the three MCK-positive samples were also found positive with PCR, however four PCR positive samples were MCK-negative. Thus, the prevalence of C.parvum was estimated as 5.2% (7/135) in our region. Of seven positive samples, one was a sewage water sample collected from the city center, while the remaining (two tap water, two well water and two sewage water samples) belonged to the samples collected from Karaduvar county, interestingly. It was thought that deficient infrastructure and use of well water as drinking water supply in Karaduvar region might be the cause of high rate of Cryptosporidium (6/45; 13.3%). Further studies which will determine the genotypes and investigate the phylogenetic relationship between these Cryptosporidium spp., might aid to the epidemiology of cryptosporidiosis in our region.

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

PubMed

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

2016-12-01

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

Survey of the Occurrence and Human Infective Potential of Giardia duodenalis and Cryptosporidium spp. in Wastewater and Different Surface Water Sources of Western Romania.

PubMed

Imre, Kálmán; Morar, Adriana; Ilie, Marius S; Plutzer, Judit; Imre, Mirela; Emil, Tîrziu; Herbei, Mihai V; Dărăbuș, Gheorghe

2017-10-01

From the group of parasitic protozoa, Giardia and Cryptosporidium are the most common pathogens spread in surface water sources, representing a continuous threat to public health and water authorities. The aim of this survey was to assess the occurrence and human infective potential of these pathogens in treated wastewaters and different surface water sources. A total of 76 western Romanian water bodies in four counties (Arad, Bihor, Caraș-Severin and Timiș) were investigated, including the effluents of wastewater treatment plants (n = 11) and brooks (n = 19), irrigation channels (n = 8), lakes (n = 16), and ponds (n = 22). Water samples were collected through polyester microfiber filtration. Giardia cysts and Cryptosporidium oocysts were isolated using immunomagnetic separation, according to the US EPA 1623 method, followed by their identification and counting by immunofluorescence (IF) microscopy. All samples were screened through PCR-based techniques targeting the gdh gene for Giardia spp. and the 18S rRNA gene for Cryptosporidium spp., followed by sequencing of the positive results. Cryptosporidium-positive samples were subtyped based on sequence analysis of the GP60 gene. Giardia spp. was found in all tested water types with a cumulative detection rate of 90.1% in wastewaters, 26.3% in brooks, 37.5% in irrigation channels, 31.2% in lakes, and 36.4% in ponds. Except for ponds, all monitored water bodies harbored the Giardia duodenalis AII subassemblage with human infective potential. In addition, the ruminant origin assemblage E was widely distributed, and the domestic/wild canid-specific assemblage D was also recorded in a pond. Three (27.3%) wastewater samples were Cryptosporidium positive, and the identified species was the zoonotic Cryptosporidium parvum, with IIaA15G2R1 (n = 2) and IIdA18G1 subtypes. The results highlight that this threat to the public health must be brought to the attention of epidemiologists, health officials, and water authorities.

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...

Sources and species of cryptosporidium oocysts in the Wachusett Reservoir watershed.

PubMed

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

2002-02-01

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

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. Copyright © 2011. Published by Elsevier Ltd.

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

PubMed

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

2017-02-01

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

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.

Monitoring of Noxious Protozoa for Management of Natural Water Resources.

PubMed

Bahk, Young Yil; Cho, Pyo Yun; Ahn, Sung Kyu; Park, Sangjung; Jheong, Won Hwa; Park, Yun-Kyu; Shin, Ho-Joon; Lee, Sang-Seob; Rhee, Okjae; Kim, Tong-Soo

2018-04-01

Waterborne parasitic protozoa, particularly Giardia lamblia and Cryptosporidium spp., are common causes of diarrhea and gastroenteritis worldwide. The most frequently identified source of infestation is water, and exposure involves either drinking water or recreation in swimming pools or natural bodies of water. In practice, studies on Cryptosporidium oocysts and Giardia cysts in surface water are challenging owing to the low concentrations of these microorganisms because of dilution. In this study, a 3-year monitoring of Cryptosporidium parvum, Giardia lamblia , and Naegleria fowleri was conducted from August 2014 to June 2016 at 5 surface water sites including 2 lakes, 1 river, and 2 water intake plants. A total of 50 water samples of 40 L were examined. Cryptosporidium oocysts were detected in 22% of samples and Giardia cysts in 32%. Water at the 5 sampling sites was all contaminated with Cryptosporidium oocysts (0-36/L), Giardia cysts (0-39/L), or both. The geometric mean concentrations of Cryptosporidium and Giardia were 1.14 oocysts/L and 4.62 cysts/L, respectively. Thus, effective monitoring plans must take into account the spatial and temporal parameters of contamination because they affect the prevalence and distribution of these protozoan cysts in local water resources.

Prevalence and genetic characterization of Cryptosporidium species and Giardia duodenalis in lambs in Oromia Special Zone, Central Ethiopia.

PubMed

Wegayehu, Teklu; Karim, Md Robiul; Li, Junqiang; Adamu, Haileeyesus; Erko, Berhanu; Zhang, Longxian; Tilahun, Getachew

2017-01-17

Cryptosporidium and Giardia duodenalis are gastro-intestinal parasites that infect human and animals worldwide. Both parasites share a broad host range and are believed to be zoonosis. The aim of this study was to identify the species of Cryptosporidium and assemblages of G. duodenalis in lambs and to elucidate their role in zoonotic transmission. A total of 389 fecal samples were collected from lambs and screened by microscopy and nested PCR targeting the small-subunit ribosomal RNA for Cryptosporidium; and the small-subunit ribosomal RNA, triose phosphate isomerase, β-giardin, and glutamate dehydrogenase genes for G. duodenalis. The prevalence of Cryptosporidium and G. duodenalis was 2.1% (8/389) and 2.6% (10/389), respectively. The infection rate at the three study sites ranged from 1.3 to 3.1% for Cryptosporidium and 1.6 to 3.9% for G. duodenalis; but variation was not statistically significant (p > 0.05). The finding also showed that there is no sex and age group associated difference in the occurrence of Cryptosporidium and G. duodenalis infections in lambs. Sequence analysis revealed that lambs were mono-infection with C. ubiquitum and G. duodenalis assemblage E. The analysis also indicated the presence of genetic variation within isolates of assemblage E; with 4 of them are novel genotypes at the small-subunit ribosomal RNA, β-giardin, and glutamate dehydrogenase genes. The findings of the current study showed that lambs are capable of harboring C. ubiquitum and G. duodenalis assemblage E. This finding suggests that lambs might be sources for potentially zoonotic Cryptosporidium species. This was first molecular study in lambs and contributes to a better understanding of the epidemiology of Cryptosporidium and G. duodenalis in central Ethiopia.

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.

Presence of Cryptosporidium spp. and Giardia duodenalis in Drinking Water Samples in the North of Portugal

PubMed Central

Moreira, Maria João; Soares, Sónia; de Lurdes Delgado, Maria; Figueiredo, João; Silva, Elisabete; Castro, António; Cosa, José Manuel Correida Da

2010-01-01

Cryptosporidium and Giardia are 2 protozoan parasites responsible for waterborne diseases outbreaks worldwide. In order to assess the prevalence of these protozoans in drinking water samples in the northern part of Portugal and the risk of human infection, we have established a long term program aiming at pinpointing the sources of surface water, drinking water, and environmental contamination, working with the water-supply industry. Total 43 sources of drinking water samples were selected, and a total of 167 samples were analyzed using the Method 1623. Sensitivity assays regarding the genetic characterization by PCR and sequencing of the genes, 18S SSU rRNA, for Cryptosporidium spp. and β,-giardin for G. duodenalis were set in the laboratory. According to the defined criteria, molecular analysis was performed over 4 samples. Environmental stages of the protozoa were detected in 25.7% (43 out of 167) of the water samples, 8.4% (14 out of 167) with cysts of Giardia, 10.2% (17 out of 167) with oocysts of Cryptosporidium and 7.2% (12 out of 167) for both species. The mean concentrations were 0.1-12.7 oocysts of Cryptosporidium spp. per 10 L and 0.1-108.3 cysts of Giardia duodenalis per 10 L. Our results suggest that the efficiency in drinking water plants must be ameliorated in their efficiency in reducing the levels of contamination. We suggest the implementation of systematic monitoring programs for both protozoa. To authors' knowledge, this is the first report evaluating the concentration of environmental stages of Cryptosporidium and Giardia in drinking water samples in the northern part of Portugal. PMID:20333284

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. Copyright © 2014 Elsevier GmbH. All rights reserved.

Assessing the infection risk of Giardia and Cryptosporidium in public drinking water delivered by surface water systems in Sao Paulo State, Brazil.

PubMed

Sato, Maria Ines Z; Galvani, Ana Tereza; Padula, Jose Antonio; Nardocci, Adelaide Cassia; Lauretto, Marcelo de Souza; Razzolini, Maria Tereza Pepe; Hachich, Elayse Maria

2013-01-01

A survey of Giardia and Cryptosporidium was conducted in surface water used as drinking water sources by public water systems in four densely urbanized regions of Sao Paulo State, Brazil. A Quantitative Microbial Risk Assessment, based on protozoa concentrations, was performed to estimate the probability of protozoa infection associated with drinking water ingestion. A total of 206 source water samples were analyzed over a 24 month period using the USEPA Method 1623. The risk of infection was estimated using an exponential dose response model, children and adults exposure and a gamma distribution for (oo)cyst concentrations with three scenarios for treating censored data. Giardia was detected in 102 of the samples, and 19 of them were also positive for Cryptosporidium, with maximum concentrations of 97.0 cysts/L and 6.0 oocysts/L, respectively. Risk distributions were similar for the three scenarios. In the four regions, the estimated risk of Giardia infection per year, for adults and children, ranged from 0.29% to 2.47% and from 0.08% to 0.70%, respectively. Cryptosporidium risk infection varied from 0.15% to 0.29% for adults and from 0.04% to 0.08% for children. In both cases, the calculated risk surpassed the risk of infection of 10(-4) (1:10,000) defined as tolerable by USEPA for a yearly exposure. The probability of Giardia infection was very close to the rates of acute diarrheic disease for adults (1% to 3%) but lower for children (2% to 7%). The daily consumption of drinking water was an important contributing factor for these differences. The Microbiological Risk Assessment carried out in this study provides an indication of infection risks by Giardia and Cryptosporidium in the population served by these source waters. Strategies for source water protection and performance targets for the water treatment should be established to achieve the required level of public health risk. Copyright © 2012 Elsevier B.V. All rights reserved.

Preventing Community-wide Transmission of Cryptosporidium: A Proactive Public Health Response to a Swimming Pool–Associated Outbreak — Auglaize County, Ohio, USA

PubMed Central

Cope, J.R.; Prosser, A.; Nowicki, S.; Roberts, M.W.; Scheer, D.; Anderson, C.; Longsworth, A.; Parsons, C.; Goldschmidt, D.; Johnston, S.; Bishop, H.; Xiao, L.; Hill, V.; Beach, M.; Hlavsa, M.C.

2015-01-01

Summary The incidence of recreational water–associated outbreaks in the United States has significantly increased, driven, at least in part, by outbreaks both caused by Cryptosporidium and associated with treated recreational water venues. Because of the parasite's extreme chlorine tolerance, transmission can occur even in well-maintained treated recreational water venues, (e.g., pools) and a focal cryptosporidiosis outbreak can evolve into a community-wide outbreak associated with multiple recreational water venues and settings (e.g., child care facilities). In August 2004 in Auglaize County, Ohio, multiple cryptosporidiosis cases were identified and anecdotally linked to Pool A. Within 5 days of the first case being reported, Pool A was hyperchlorinated to achieve 99.9% Cryptosporidium inactivition. A case-control study was launched to epidemiologically ascertain the outbreak source 11 days later. A total of 150 confirmed and probable cases were identified; the temporal distribution of illness onset was peaked, indicating a point-source exposure. Cryptosporidiosis was significantly associated with swimming in Pool A (matched odds ratio 121.7, 95% confidence interval 27.4–∞) but not with another venue or setting. The findings of this investigation suggest that proactive implementation of control measures, when increased Cryptosporidium transmission is detected but before an outbreak source is epidemiologically ascertained, might prevent a focal cryptosporidiosis outbreak from evolving into a community-wide outbreak. PMID:25907106

77 FR 57545 - Long Term 2 Enhanced Surface Water Treatment Rule: Public Meeting on Monitoring Data Analysis...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-09-18

... on the occurrence of Cryptosporidium or E. coli in their source waters. Systems that are placed into... categories (i.e., bins) a public drinking water system (PWS) should be placed. The second topic is the... of Escherichia coli as a screen to identify small filtered PWSs that need to perform Cryptosporidium...

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

PubMed

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

2016-10-01

This study describes cryptosporidiosis in an overwintering group of 15 European hedgehogs (Erinaceus europaeus), comprising 3 adults and 12 juveniles. Four juvenile hedgehogs were hospitalised with anorexia, malodorous diarrhoea and dehydration. Immediate parasitological examinations revealed the presence of Cryptosporidium sp. in these animals and also in 5 other juveniles. All hedgehogs were coproscopically monitored for 4 months over the winter season. Shedding of Cryptosporidium oocysts persisted from 6 to 70 days. Repeated shedding of Cryptosporidium oocysts occurred in 3 animals after 4 months subsequent to the first outbreak. Clinical signs were observed only at the beginning of the outbreak (apathy, anorexia, general weakness, mild dehydration, and malodorous faeces with changed consistence - soft/diarrhoea) in the 4 hospitalised juveniles. Overall 11 hedgehogs were Cryptosporidium-positive, both microscopically and by PCR methods. Sequence analyses of SSU rRNA and gp60 genes revealed the presence of C. parvum IIdA18G1 subtype in all positive hedgehogs. Moreover, 3 hedgehogs had a mixed infection of the zoonotic C. parvum and C. erinacei XIIIaA19R13 subtype. Cryptosporidium infections can be rapidly spread among debilitated animals and the positive hedgehogs released back into the wild can be a source of the infection for individuals weakened after hibernation. Copyright © 2016 Elsevier GmbH. All rights reserved.

Spinacia oleracea L. Leaf Stomata Harboring Cryptosporidium parvum Oocysts: a Potential Threat to Food Safety ▿ †

PubMed Central

Macarisin, Dumitru; Bauchan, Gary; Fayer, Ronald

2010-01-01

Cryptosporidium parvum is a cosmopolitan microscopic protozoan parasite that causes severe diarrheal disease (cryptosporidiosis) in mammals, including humans and livestock. There is growing evidence of Cryptosporidium persistence in fresh produce that may result in food-borne infection, including sporadic cases as well as outbreaks. However, drinking and recreational waters are still considered the major sources of Cryptosporidium infection in humans, which has resulted in prioritization of studies of parasite etiology in aquatic environments, while the mechanisms of transmission and parasite persistence on edible plants remain poorly understood. Using laser scanning confocal microscopy together with fluorescein-labeled monoclonal antibodies, C. parvum oocysts were found to strongly adhere to spinach plants after contact with contaminated water, to infiltrate through the stomatal openings in spinach leaves, and to persist at the mesophyll level. These findings and the fact that this pathogenic parasite resists washing and disinfection raise concerns regarding food safety. PMID:19933348

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.

Influence of land use and watershed characteristics on protozoa contamination in a potential drinking water resources reservoir.

PubMed

Keeley, Ann; Faulkner, Barton R

2008-05-01

Relative changes in the microbial quality of Lake Texoma, on the border of Texas and Oklahoma, were investigated by monitoring protozoan pathogens, fecal indicators, and factors influencing the intensity of the microbiological contamination of surface water reservoirs. The watershed serves rural agricultural communities active in cattle ranching, recreation, and is a potential drinking water source. A total of 193 surface water samples were tested over a 27-month period to determine levels of parasite contamination. The overall occurrence of Cryptosporidium oocysts was higher in both frequency and concentration than Giardia cysts. Cryptosporidium oocysts were found in 99% and Giardia cysts in 87% of the samples. Although Cryptosporidium and Giardia occurrence were significantly but not strongly correlated, all other correlation coefficients including turbidity and total dissolved solids were non-significant. Statistically supportable seasonal variations were found suggesting that Cryptosporidium and Giardia were higher in summer and fall than in other seasons of the year. While Cryptosporidium levels were correlated with rainfall, this was not the case with Giardia. The maximum numbers for both protozoan parasites were detected from a site impacted by cattle ranching during calving season. Restriction fragment length polymorphism analysis was used for confirmation of Cryptosporidium in surface waters influenced by agricultural discharges. As we had expected, oocysts were of the bovine type indicating that the Cryptosporidium parvum detected in surface waters perhaps came from cattle living in the watershed.

Prevalence and molecular characterization of Cryptosporidium spp. and Giardia duodenalis in deer in Henan and Jilin, China.

PubMed

Huang, Jianying; Zhang, Zhenjie; Zhang, Yiqi; Yang, Yong; Zhao, Jinfeng; Wang, Rongjun; Jian, Fuchun; Ning, Changshen; Zhang, Wanyu; Zhang, Longxian

2018-04-12

Little is known about the prevalence and zoonotic potential of Cryptosporidium spp. and Giardia duodenalis in deer in China. In this study, 662 fecal samples were collected from 11 farms in Henan and Jilin Provinces between July 2013 and August 2014, and were screened for the presence of Cryptosporidium and G. duodenalis with genotyping and subtyping methods. Cryptosporidium spp. and G. duodenalis were detected in 6.80% (45/662) and 1.21% (5/662) of samples, respectively. Six Cryptosporidium species/genotypes were identified based on the small subunit ribosomal ribonucleic acid (SSU rRNA) gene: C. parvum (n = 11); C. andersoni (n = 5); C. ubiquitum (n = 3); C. muris (n = 1); C. suis-like (n = 1); and Cryptosporidium deer genotype (n = 24). When five of the 11 C. parvum isolates were subtyped by sequencing the 60 kDa glycoprotein (gp60) gene, zoonotic subtypes IIaA15G2R2 (n = 4) and IIdA19G1 (n = 1) were found. According to a subtype analysis, three C. ubiquitum isolates belonged to XIIa subtype 2. In contrast, only assemblage E was detected in the five Giardia-positive samples with small subunit ribosomal ribonucleic acid (SSU rRNA) gene sequencing. To our knowledge, this is the first study to report C. andersoni, as well as C. parvum zoonotic subtypes IIaA15G2R2 and IIdA19G1 in cervids. These data, though limited, suggest that cervids may be a source of zoonotic Cryptosporidium and Giardia. Cervids in the present study are likely to be of low zoonotic potential to humans, and more molecular epidemiological studies are required to clarify the prevalence and public health significance of Cryptosporidium and G. duodenalis in cervids throughout China.

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kelley, M.B.; Brokaw, J.K.; Brokaw, J.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 helpfulmore » 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.« less

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.

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

PubMed Central

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

2015-01-01

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

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.

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.

40 CFR 141.701 - Source water monitoring.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 23 2011-07-01 2011-07-01 false Source water monitoring. 141.701 Section 141.701 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Source Water...

40 CFR 141.701 - Source water monitoring.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 22 2010-07-01 2010-07-01 false Source water monitoring. 141.701 Section 141.701 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Source Water...

40 CFR 141.701 - Source water monitoring.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 24 2012-07-01 2012-07-01 false Source water monitoring. 141.701 Section 141.701 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Source Water...

40 CFR 141.701 - Source water monitoring.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 24 2013-07-01 2013-07-01 false Source water monitoring. 141.701 Section 141.701 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Source Water...

40 CFR 141.701 - Source water monitoring.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 23 2014-07-01 2014-07-01 false Source water monitoring. 141.701 Section 141.701 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Source Water...

40 CFR 141.706 - Reporting source water monitoring results.

Code of Federal Regulations, 2013 CFR

2013-07-01

... results. 141.706 Section 141.706 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... Cryptosporidium Source Water Monitoring Requirements § 141.706 Reporting source water monitoring results. (a) Systems must report results from the source water monitoring required under § 141.701 no later than 10...

40 CFR 141.706 - Reporting source water monitoring results.

Code of Federal Regulations, 2012 CFR

2012-07-01

... results. 141.706 Section 141.706 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... Cryptosporidium Source Water Monitoring Requirements § 141.706 Reporting source water monitoring results. (a) Systems must report results from the source water monitoring required under § 141.701 no later than 10...

Occurrence and molecular characterization of Cryptosporidium spp. isolated from domestic animals in a rural area surrounding Atlantic dry forest fragments in Teodoro Sampaio municipality, State of São Paulo, Brazil.

PubMed

Sevá, Anaiá da Paixão; Funada, Mikaela Renata; Souza, Sheila de Oliveira; Nava, Alessandra; Richtzenhain, Leonardo José; Soares, Rodrigo Martins

2010-01-01

The aim of this study was to assess the occurrence of Cryptosporidium in domestic animals in rural properties surrounding rain forest fragments within the municipality of Teodoro Sampaio, southeastern Brazil. Conventional sucrose flotation method followed by molecular characterization of the parasites by sequencing PCR products amplified from SSU rRNA gene were used. Stool samples were collected from domestic animals raised as pets and livestock in all rural properties surrounding three forest fragments. Samples from cattle (197), equine (63), pigs (25), sheep (11), and dogs (28) were collected from 98 rural properties. The frequency of occurrence of Cryptosporidium within each animal species was 3.0% (6/197) among cattle and 10.7% (3/28) among dogs. Cryptosporidium was not detected in stool samples from equine, sheep, and pigs. All sequences obtained from the six samples of calves showed molecular identity with Cryptosporidium andersoni while all sequences from dog samples were similar to C. canis. The frequency of occurrence of Cryptosporidium in these domestic animal species was low. The absence of C. parvum in the present study suggests that the zoonotic cycle of cryptosporidiosis may not be relevant in the region studied. The presence of Cryptosporidium species seldom described in humans may be, otherwise, important for the wild fauna as these animals are a source of infection and dissemination of this protozoan to other animal species. The impact and magnitude of infection by C. andersoni in wild ruminants and C. canis in wild canids have to be assessed in future studies to better understand the actual importance of these species in this region.

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.

Cryptosporidium spp. and Giardia spp. in feces and water and the associated exposure factors on dairy farms

PubMed Central

Martins, Felippe Danyel Cardoso; Ferreira, Fernanda Pinto; de Almeida, Jonatas Campos; Ogawa, Liza; dos Santos, Hannah Lia Ettiene Peruch Lemos; dos Santos, Maíra Moreira; Pinheiro, Filipe Aguera; Navarro, Italmar Teodorico; Garcia, João Luis; Freire, Roberta Lemos

2017-01-01

The aims of this study were to verify the prevalence of Cryptosporidium spp. and Giardia spp. in animal feces and drinking water on dairy farms and to identify a possible relation between the exposure factors and the presence of these parasites. Fecal samples from cattle and humans and water samples were collected on dairy farms in Paraná, Brazil. Analysis of (oo)cysts in the feces was performed by the modified Ziehl-Neelsen staining and centrifugal flotation in zinc sulfate. Test-positive samples were subjected to nested PCR amplification of the 18SSU ribosomal RNA gene for identification of Cryptosporidium and Giardia and of the gp60 gene for subtyping of Cryptosporidium. Microbiological analysis of water was carried out by the multiple-tube method and by means of a chromogenic substrate, and parasitological analysis was performed on 31 samples by direct immunofluorescence and nested PCR of the genes mentioned above. Identification of the species of Cryptosporidium was performed by sequencing and PCR with analysis of restriction fragment length polymorphisms. The prevalence of Giardia and Cryptosporidium was higher in calves than in adults. Among the samples of cattle feces, Cryptosporidium parvum was identified in 41 (64%), C. ryanae in eight (12.5%), C. bovis in four (6.3%), C. andersoni in five (7.8%), and a mixed infection in 20 samples (31.3%). These parasites were not identified in the samples of human feces. Thermotolerant coliform bacteria were identified in 25 samples of water (45.5%). Giardia duodenalis and C. parvum were identified in three water samples. The gp60 gene analysis of C. parvum isolates revealed the presence of two strains (IIaA20G1R1 and IIaA17G2R2) in the fecal samples and one (IIaA17G2R1) in the water samples. The presence of coliforms was associated with the water source, structure and degradation of springs, rain, and turbidity. The prevalence of protozoa was higher in calves up to six months of age. C. parvum and G. duodenalis were identified in the water of dairy farms, as were thermotolerant coliforms; these findings point to the need for guidance on handling of animals, preservation of water sources, and water treatment. PMID:28403147

Cryptosporidium spp. and Giardia spp. in feces and water and the associated exposure factors on dairy farms.

PubMed

Toledo, Roberta Dos Santos; Martins, Felippe Danyel Cardoso; Ferreira, Fernanda Pinto; de Almeida, Jonatas Campos; Ogawa, Liza; Dos Santos, Hannah Lia Ettiene Peruch Lemos; Dos Santos, Maíra Moreira; Pinheiro, Filipe Aguera; Navarro, Italmar Teodorico; Garcia, João Luis; Freire, Roberta Lemos

2017-01-01

The aims of this study were to verify the prevalence of Cryptosporidium spp. and Giardia spp. in animal feces and drinking water on dairy farms and to identify a possible relation between the exposure factors and the presence of these parasites. Fecal samples from cattle and humans and water samples were collected on dairy farms in Paraná, Brazil. Analysis of (oo)cysts in the feces was performed by the modified Ziehl-Neelsen staining and centrifugal flotation in zinc sulfate. Test-positive samples were subjected to nested PCR amplification of the 18SSU ribosomal RNA gene for identification of Cryptosporidium and Giardia and of the gp60 gene for subtyping of Cryptosporidium. Microbiological analysis of water was carried out by the multiple-tube method and by means of a chromogenic substrate, and parasitological analysis was performed on 31 samples by direct immunofluorescence and nested PCR of the genes mentioned above. Identification of the species of Cryptosporidium was performed by sequencing and PCR with analysis of restriction fragment length polymorphisms. The prevalence of Giardia and Cryptosporidium was higher in calves than in adults. Among the samples of cattle feces, Cryptosporidium parvum was identified in 41 (64%), C. ryanae in eight (12.5%), C. bovis in four (6.3%), C. andersoni in five (7.8%), and a mixed infection in 20 samples (31.3%). These parasites were not identified in the samples of human feces. Thermotolerant coliform bacteria were identified in 25 samples of water (45.5%). Giardia duodenalis and C. parvum were identified in three water samples. The gp60 gene analysis of C. parvum isolates revealed the presence of two strains (IIaA20G1R1 and IIaA17G2R2) in the fecal samples and one (IIaA17G2R1) in the water samples. The presence of coliforms was associated with the water source, structure and degradation of springs, rain, and turbidity. The prevalence of protozoa was higher in calves up to six months of age. C. parvum and G. duodenalis were identified in the water of dairy farms, as were thermotolerant coliforms; these findings point to the need for guidance on handling of animals, preservation of water sources, and water treatment.

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. © The American Society of Tropical Medicine and Hygiene.

Recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for equipment-free detection of Cryptosporidium spp. oocysts in dairy cattle feces.

PubMed

Wu, Yao-Dong; Zhou, Dong-Hui; Zhang, Long-Xian; Zheng, Wen-Bin; Ma, Jian-Gang; Wang, Meng; Zhu, Xing-Quan; Xu, Min-Jun

2016-09-01

Cryptosporidium is a widespread protozoan parasite that infects a large number of vertebrate animals, resulting in varying degrees of diarrhea or even death. As dairy cattle feces is an important source of Cryptosporidium spp. infection, development of a handy and accurate detection method via its oocysts in dairy cattle feces would be interesting and necessary. We herein developed a quick detecting method using recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip to detect DNA of Cryptosporidium oocysts in dairy cattle feces. The DNA was released by boiled water with 0.1 % N-lauroylsarcosine sodium salt (LSS). The established method was proven to be of higher sensitivity than normal polymerase chain reaction (PCR) amplification with the lowest detection of 0.5 oocyst per reaction, and specificity with no cross reactivity to other common protozoan species in the intestine of dairy cattle. The diagnostic method established herein is simple, rapid, and cost-effective, and has potential for further development as a diagnostic kit for the diagnosis of cryptosporidiosis of dairy cattle.

40 CFR 141.706 - Reporting source water monitoring results.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 23 2014-07-01 2014-07-01 false Reporting source water monitoring results. 141.706 Section 141.706 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Source Water Monitoring Requirements §...

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.

Prevalence and Molecular Characterization of Cryptosporidium in Goats across Four Provincial Level Areas in China

PubMed Central

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. PMID:25343501

Microbial health risks associated with exposure to stormwater in a water plaza.

PubMed

Sales-Ortells, Helena; Medema, Gertjan

2015-05-01

Climate change scenarios predict an increase of intense rainfall events in summer in Western Europe. Current urban drainage systems cannot cope with such intense precipitation events. Cities are constructing stormwater storage facilities to prevent pluvial flooding. Combining storage with other functions, such as recreation, may lead to exposure to contaminants. This study assessed the microbial quality of rainwater collected in a water plaza and the health risks associated with recreational exposure. The water plaza collects street run-off, diverges first flush to the sewer system and stores the rest of the run-off in the plaza as open water. Campylobacter, Cryptosporidium and Legionella pneumophila were the pathogens investigated. Microbial source tracking tools were used to determine the origin (human, animal) of the intestinal pathogens. Cryptosporidium was not found in any sample. Campylobacter was found in all samples, with higher concentrations in samples containing human Bacteroides than in samples with zoonotic contamination (15 vs 3.7 gc (genomic copies)/100 mL). In both cases, the estimated disease risk associated with Campylobacter and recreational exposure was higher than the Dutch national incidence. This indicates that the health risk associated with recreational exposure to the water plaza is significant. L. pneumophila was found only in two out of ten pond samples. Legionnaire's disease risks were lower than the Dutch national incidence. Presence of human Bacteroides indicates possible cross-connections with the CSS that should be identified and removed. Copyright © 2015 Elsevier Ltd. All rights reserved.

Transport of Cryptosporidium parvum Oocysts in Charge Heterogeneous Porous Media: Microfluidics Experiment and Numerical Simulation

NASA Astrophysics Data System (ADS)

Liu, Y.; Meng, X.; Guo, Z.; Zhang, C.; Nguyen, T. H.; Hu, D.; Ji, J.; Yang, X.

2017-12-01

Colloidal attachment on charge heterogeneous grains has significant environmental implications for transport of hazardous colloids, such as pathogens, in the aquifer, where iron, manganese, and aluminium oxide minerals are the major source of surface charge heterogeneity of the aquifer grains. A patchwise surface charge model is often used to describe the surface charge heterogeneity of the grains. In the patchwise model, the colloidal attachment efficiency is linearly correlated with the fraction of the favorable patches (θ=λ(θf - θu)+θu). However, our previous microfluidic study showed that the attachment efficiency of oocysts of Cryptosporidium parvum, a waterborne protozoan parasite, was not linear correlated with the fraction of the favorable patches (λ). In this study, we developed a pore scale model to simulate colloidal transport and attachment on charge heterogeneous grains. The flow field was simulated using the LBM method and colloidal transport and attachment were simulated using the Lagrange particle tracking method. The pore scale model was calibrated with experimental results of colloidal and oocyst transport in microfluidic devices and was then used to simulate oocyst transport in charge heterogeneous porous media under a variety of environmental relative conditions, i.e. the fraction of favorable patchwise, ionic strength, and pH. The results of the pore scale simulations were used to evaluate the effect of surface charge heterogeneity on upscaling of oocyst transport from pore to continuum scale and to develop an applicable correlation between colloidal attachment efficiency and the fraction of the favorable patches.

Cryptosporidium and Giardia in Africa: current and future challenges.

PubMed

Squire, Sylvia Afriyie; Ryan, Una

2017-04-20

Cryptosporidium and Giardia are important causes of diarrhoeal illness. Adequate knowledge of the molecular diversity and geographical distribution of these parasites and the environmental and climatic variables that influence their prevalence is important for effective control of infection in at-risk populations, yet relatively little is known about the epidemiology of these parasites in Africa. Cryptosporidium is associated with moderate to severe diarrhoea and increased mortality in African countries and both parasites negatively affect child growth and development. Malnutrition and HIV status are also important contributors to the prevalence of Cryptosporidium and Giardia in African countries. Molecular typing of both parasites in humans, domestic animals and wildlife to date indicates a complex picture of both anthroponotic, zoonotic and spill-back transmission cycles that requires further investigation. For Cryptosporidium, the only available drug (nitazoxanide) is ineffective in HIV and malnourished individuals and therefore more effective drugs are a high priority. Several classes of drugs with good efficacy exist for Giardia, but dosing regimens are suboptimal and emerging resistance threatens clinical utility. Climate change and population growth are also predicted to increase both malnutrition and the prevalence of these parasites in water sources. Dedicated and co-ordinated commitments from African governments involving "One Health" initiatives with multidisciplinary teams of veterinarians, medical workers, relevant government authorities, and public health specialists working together are essential to control and prevent the burden of disease caused by these parasites.

Zoonotic Cryptosporidium Species in Animals Inhabiting Sydney Water Catchments

PubMed Central

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

2016-01-01

Cryptosporidium is one of the most common zoonotic waterborne parasitic diseases worldwide and represents a major public health concern of water utilities in developed nations. As animals in catchments can shed human-infectious Cryptosporidium oocysts, determining the potential role of animals in dissemination of zoonotic Cryptosporidium to drinking water sources is crucial. In the present study, a total of 952 animal faecal samples from four dominant species (kangaroos, rabbits, cattle and sheep) inhabiting Sydney’s drinking water catchments were screened for the presence of Cryptosporidium using a quantitative PCR (qPCR) and positives sequenced at multiple loci. Cryptosporidium species were detected in 3.6% (21/576) of kangaroos, 7.0% (10/142) of cattle, 2.3% (3/128) of sheep and 13.2% (14/106) of rabbit samples screened. Sequence analysis of a region of the 18S rRNA locus identified C. macropodum and C. hominis in 4 and 17 isolates from kangaroos respectively, C. hominis and C. parvum in 6 and 4 isolates respectively each from cattle, C. ubiquitum in 3 isolates from sheep and C. cuniculus in 14 isolates from rabbits. All the Cryptosporidium species identified were zoonotic species with the exception of C. macropodum. Subtyping using the 5’ half of gp60 identified C. hominis IbA10G2 (n = 12) and IdA15G1 (n = 2) in kangaroo faecal samples; C. hominis IbA10G2 (n = 4) and C. parvum IIaA18G3R1 (n = 4) in cattle faecal samples, C. ubiquitum subtype XIIa (n = 1) in sheep and C. cuniculus VbA23 (n = 9) in rabbits. Additional analysis of a subset of samples using primers targeting conserved regions of the MIC1 gene and the 3’ end of gp60 suggests that the C. hominis detected in these animals represent substantial variants that failed to amplify as expected. The significance of this finding requires further investigation but might be reflective of the ability of this C. hominis variant to infect animals. The finding of zoonotic Cryptosporidium species in these animals may have important implications for the management of drinking water catchments to minimize risk to public health. PMID:27973572

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

Prevalence of Cryptosporidium spp., Enterocytozoon bieneusi, Encephalitozoon spp. and Giardia intestinalis in Wild, Semi-Wild and Captive Orangutans (Pongo abelii and Pongo pygmaeus) on Sumatra and Borneo, Indonesia.

PubMed

Mynářová, Anna; Foitová, Ivona; Kváč, Martin; Květoňová, Dana; Rost, Michael; Morrogh-Bernard, Helen; Nurcahyo, Wisnu; Nguyen, Cathleen; Supriyadi, Supriyadi; Sak, Bohumil

2016-01-01

Orangutans are critically endangered primarily due to loss and fragmentation of their natural habitat. This could bring them into closer contact with humans and increase the risk of zoonotic pathogen transmission. To describe the prevalence and diversity of Cryptosporidium spp., microsporidia and Giardia intestinalis in orangutans at seven sites on Sumatra and Kalimantan, and to evaluate the impact of orangutans' habituation and location on the occurrence of these zoonotic protists. The overall prevalence of parasites in 298 examined animals was 11.1%. The most prevalent microsporidia was Encephalitozoon cuniculi genotype II, found in 21 animals (7.0%). Enterocytozoon bieneusi genotype D (n = 5) and novel genotype Pongo 2 were detected only in six individuals (2.0%). To the best of our knowledge, this is the first report of these parasites in orangutans. Eight animals were positive for Cryptosporidium spp. (2.7%), including C. parvum (n = 2) and C. muris (n = 6). Giardia intestinalis assemblage B, subtype MB6, was identified in a single individual. While no significant differences between the different human contact level groups (p = 0.479-0.670) or between the different islands (p = 0.992) were reported in case of E. bieneusi or E. cuniculi, Cryptosporidium spp. was significantly less frequently detected in wild individuals (p < 2×10-16) and was significantly more prevalent in orangutans on Kalimantan than on Sumatra (p < 2×10-16). Our results revealed that wild orangutans are significantly less frequently infected by Cryptosporidium spp. than captive and semi-wild animals. In addition, this parasite was more frequently detected at localities on Kalimantan. In contrast, we did not detect any significant difference in the prevalence of microsporidia between the studied groups of animals. The sources and transmission modes of infections were not determined, as this would require repeated sampling of individuals, examination of water sources, and sampling of humans and animals sharing the habitat with orangutans.

Characterizing the concentration of Cryptosporidium in Australian surface waters for setting health-based targets for drinking water treatment.

PubMed

Petterson, S; Roser, D; Deere, D

2015-09-01

It is proposed that the next revision of the Australian Drinking Water Guidelines will include 'health-based targets', where the required level of potable water treatment quantitatively relates to the magnitude of source water pathogen concentrations. To quantify likely Cryptosporidium concentrations in southern Australian surface source waters, the databases for 25 metropolitan water supplies with good historical records, representing a range of catchment sizes, land use and climatic regions were mined. The distributions and uncertainty intervals for Cryptosporidium concentrations were characterized for each site. Then, treatment targets were quantified applying the framework recommended in the World Health Organization Guidelines for Drinking-Water Quality 2011. Based on total oocyst concentrations, and not factoring in genotype or physiological state information as it relates to infectivity for humans, the best estimates of the required level of treatment, expressed as log10 reduction values, ranged among the study sites from 1.4 to 6.1 log10. Challenges associated with relying on historical monitoring data for defining drinking water treatment requirements were identified. In addition, the importance of quantitative microbial risk assessment input assumptions on the quantified treatment targets was investigated, highlighting the need for selection of locally appropriate values.

Water used to moisten vegetables is a source of Escherichia coli and protozoan parasite contamination at markets in Hanoi, Vietnam.

PubMed

Tram, Nguyen Thuy; Dalsgaard, Anders

2014-12-01

The study was done to assess the level of fecal (Escherichia coli) and protozoan parasite (Cryptosporidium spp. and Giardia spp.) contamination in water used by traders to moisten vegetables at markets in Hanoi, Vietnam. A total of 200 splashing water samples from markets located within eight districts were analyzed for E. coli and Cryptosporidium spp. and Giardia spp. (oo)cysts. Giardia cysts were found in 17 splashing water samples and Cryptosporidium oocysts in nine samples, with median values of 20 cysts ml(-1) and 10 oocysts ml(-1), respectively. E. coli was found with a median concentration of 636 cfu ml(-1) and its occurrence was negatively correlated with the numbers of protozoan parasites. The splashing water was kept in buckets that were rarely cleaned and often used for handwashing. The finding of these pathogens in splashing water is likely to represent real food safety hazards.

40 CFR 141.716 - Source toolbox components.

Code of Federal Regulations, 2012 CFR

2012-07-01

... for Microbial Toolbox Components § 141.716 Source toolbox components. (a) Watershed control program. Systems receive 0.5-log Cryptosporidium treatment credit for implementing a watershed control program that meets the requirements of this section. (1) Systems that intend to apply for the watershed control...

40 CFR 141.716 - Source toolbox components.

Code of Federal Regulations, 2013 CFR

2013-07-01

... for Microbial Toolbox Components § 141.716 Source toolbox components. (a) Watershed control program. Systems receive 0.5-log Cryptosporidium treatment credit for implementing a watershed control program that meets the requirements of this section. (1) Systems that intend to apply for the watershed control...

40 CFR 141.716 - Source toolbox components.

Code of Federal Regulations, 2014 CFR

2014-07-01

... for Microbial Toolbox Components § 141.716 Source toolbox components. (a) Watershed control program. Systems receive 0.5-log Cryptosporidium treatment credit for implementing a watershed control program that meets the requirements of this section. (1) Systems that intend to apply for the watershed control...

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.

Genetic Diversity of Cryptosporidium spp. in Captive Reptiles

PubMed Central

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

2004-01-01

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

A survey for potentially zoonotic gastrointestinal parasites in domestic cavies in Cameroon (Central Africa).

PubMed

Meutchieye, Felix; Kouam, Marc K; Miegoué, Emile; Nguafack, Terence T; Tchoumboué, Joseph; Téguia, Alexis; Théodoropoulos, Georgios

2017-06-26

Farm animals are usually suspected to transmit infections to humans. Domestic cavies (Cavia porcellus) are hosts to a variety of pathogens, some of which are zoonotic. Several parasites including the protozoa Giardia spp. and Cryptosporidium spp. may be causative agents of gastrointestinal disorders in domestic cavies and humans. The aim of the study was to investigate the occurrence of potentially zoonotic protozoa as well as any potential zoonotic gastrointestinal parasite in domestic cavies raised under a semi extensive system in the rural areas of Cameroon. Giardia/Cryptosporidium antigens were detected in 12.90% of cavies. Helminthe eggs were found in 1.52% of animals. The prevalence of Paraspidodera uncinata, Heligmosomoides polygyrus (also known as Nematospiroides dubius) and Trichuris sp. was 1% (4/397), 0.3% (1/397), and 0.3% (1/397), respectively. Presence of Giardia/Cryptosporidium was unrelated to the occurrence of diarrhea, as none of the positive samples was from a diarrheic individual. Domestic cavies are hosts of Giardia/Cryptosporidium and appear as potential source of human giardiasis, cryptosporidiosis and infection with H. polygyrus in Cameroon. In keeping with the One Health Initiative, veterinarians and medical doctors should collaborate to address the problem of Giardia and Cryptosporidium infection in cavies and cavy breeders both in Cameroon and other countries with a similar cavy breeding system. Follow-up studies are required to further taxonomically characterize these cavy parasites and to determine their routes of transmission to humans.

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 Conclusions GloWPa-Crypto is the first global model that can be used to analyse dynamics in surface water pathogen concentrations worldwide. Global human Cryptosporidium emissions are estimated at 1 x 10^17 oocysts/ year for the year 2010.We estimated future emissions for SSP1 and SSP3. Preliminary results show that for SSP1human emissions are approximately halved by 2050. The SSP3 human emissions are 1.5 times higher than the 2010 emissions due to increased population growth and urbanisation. Livestock Cryptosporidium emissions are expected to increase under both SSP1 and SSP3, as meat consumption continues to rise. We conclude that population growth, urbanization, changes in sanitation systems and treatment, and changes in livestock consumption and production systems are important processes that determine future Cryptosporidium emissions to surface water. References Hofstra N, Bouwman A F, Beusen A H W and Medema G J 2013 Exploring global Cryptosporidium emissions to surface water Sci. Total Environ. 442 10-9 Kiulia N M, Hofstra N, Vermeulen L C, Obara M A, Medema G J and Rose J B 2015 Global occurrence and emission of rotaviruses to surface waters Pathogens 4 229-55 Vermeulen L C, De Kraker J, Hofstra N, Kroeze C and Medema G J 2015 Modelling the impact of sanitation, population and urbanization estimates on human emissions of Cryptosporidium to surface waters - a case study for Bangladesh and India Environ. Res. Lett. 10

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. © 2013 Elsevier Ireland Ltd. All rights reserved.

Detection and molecular identification of Cryptosporidium species in laboratory rats (Rattus norvegicus) in Ibadan, Nigeria

PubMed

Ayinmode, Adekunle Bamidele; Ogbonna, Nkeiruka Fortunate; Widmer, Giovanni

To study the occurrence of Cryptosporidium infection in laboratory rats (Rattus norvegicus) raised for experimental usage, 134 faecal samples were obtained from two rearing houses in Ibadan and examined for the presence of Cryptosporidium oocyst using the modified acid fast staining technique. Cryptosporidium species in 2 samples positive for microscopy were further characterized by a nested polymerase chain reaction (PCR) amplifying the 18S rRNA gene. Two of 134 samples were positive for the Cryptosporidium oocysts. Sequencing of the small-subunit rRNA amplicons identified the species in the two PCR positive samples as Cryptosporidium andersoni and Cryptosporidium rat genotype. These findings showed that laboratory rat is a potential reservoir for diverse Cryptosporidium species and suggests that laboratory rats should be screened for Cryptosporidium infection prior to experiments, especially where pathogen free animals are not available. This the first report to identify Cryptosporidium species infecting laboratory rats in Nigeria.

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.

Microbial pathogens in source and treated waters from drinking water treatment plants in the US

EPA Science Inventory

An occurrence survey was conducted on selected pathogens in source and treated drinking water collected from 25 drinking water treatment plants (DWTPs) in the United States. Water samples were analyzed for the protozoa Giardia and Cryptosporidium (EPA Method 1623); the fungi Asp...

40 CFR 141.717 - Pre-filtration treatment toolbox components.

Code of Federal Regulations, 2011 CFR

2011-07-01

... surface water or GWUDI source. (c) Bank filtration. Systems receive Cryptosporidium treatment credit for... paragraph. Systems using bank filtration when they begin source water monitoring under § 141.701(a) must... 40 Protection of Environment 23 2011-07-01 2011-07-01 false Pre-filtration treatment toolbox...

Differences in staining intensities affect reported occurrences and concentrations of Giardia spp. in surface drinking water sources

EPA Science Inventory

Aim USEPA Method 1623, or its equivalent, is currently used to monitor for protozoan contamination of surface drinking water sources worldwide. At least three approved staining kits used for detecting Cryptosporidium and Giardia are commercially available. This study focuses on ...

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. Copyright © 2013 Elsevier Ltd. All rights reserved.

Assessment of groundwater under direct influence of surface water.

PubMed

Nnadi, Fidelia N; Fulkerson, Mark

2002-08-01

Waterborne pathogens are known to reside in surface water systems throughout the U.S. Cryptosporidium outbreaks over recent years are the result of drinking water supplied from such sources. Contamination of aquifers has also led to several reported cases from drinking water wells. With high resistance to typical groundwater treatment procedures, aquifer infiltration by Cryptosporidium poses a serious threat. As groundwater wells are the main source of drinking water supply in the State of Florida, understanding factors that affect the presence of Cryptosporidium would prevent future outbreaks. This study examines karst geology, land use, and hydrogeology in the State of Florida as they influence the risk of groundwater contamination. Microscopic Particulate Analysis (MPA) sampling was performed on 719 wells distributed across Florida. The results of the sampling described each well as having high, moderate, or low risk to surface water influence. The results of this study indicated that the hydrogeology of an area tends to influence the MPA Risk Index (RI) of a well. Certain geologic formations were present for the majority of the high risk wells. Residential land use contained nearly half of the wells sampled. The results also suggested that areas more prone to sinkhole development are likely to contain wells with a positive RI.

Communitywide cryptosporidiosis outbreak associated with a surface water-supplied municipal water system--Baker City, Oregon, 2013.

PubMed

DeSilva, M B; Schafer, S; Kendall Scott, M; Robinson, B; Hills, A; Buser, G L; Salis, K; Gargano, J; Yoder, J; Hill, V; Xiao, L; Roellig, D; Hedberg, K

2016-01-01

Cryptosporidium, a parasite known to cause large drinking and recreational water outbreaks, is tolerant of chlorine concentrations used for drinking water treatment. Human laboratory-based surveillance for enteric pathogens detected a cryptosporidiosis outbreak in Baker City, Oregon during July 2013 associated with municipal drinking water. Objectives of the investigation were to confirm the outbreak source and assess outbreak extent. The watershed was inspected and city water was tested for contamination. To determine the community attack rate, a standardized questionnaire was administered to randomly sampled households. Weighted attack rates and confidence intervals (CIs) were calculated. Water samples tested positive for Cryptosporidium species; a Cryptosporidium parvum subtype common in cattle was detected in human stool specimens. Cattle were observed grazing along watershed borders; cattle faeces were observed within watershed barriers. The city water treatment facility chlorinated, but did not filter, water. The community attack rate was 28·3% (95% CI 22·1-33·6), sickening an estimated 2780 persons. Watershed contamination by cattle probably caused this outbreak; water treatments effective against Cryptosporidium were not in place. This outbreak highlights vulnerability of drinking water systems to pathogen contamination and underscores the need for communities to invest in system improvements to maintain multiple barriers to drinking water contamination.

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

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. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Detection by PCR of pathogenic protozoa in raw and drinkable water samples in Colombia.

PubMed

Triviño-Valencia, Jessica; Lora, Fabiana; Zuluaga, Juan David; Gomez-Marin, Jorge E

2016-05-01

We evaluated the presence of DNA of Giardia, Toxoplasma, and Cryptosporidium by PCR, and of Giardia and Cryptosporidium genera by immunofluorescence antibody test (IFAT), in water samples, before, during, and after plant treatment for drinkable water. We applied this method in 38 samples of 10 l of water taken from each of the water treatment steps and in 8 samples taken at home (only for Toxoplasma PCR) in Quindio region in Colombia. There were 8 positive samples for Cryptosporidium parvum (21 %), 4 for Cryptosporidium hominis (10.5 %), 27 for Toxoplasma gondii (58.6 %), 2 for Giardia duodenalis assemblage A (5.2 %), and 5 for G. duodenalis assemblage B (13.1 %). By IFAT, 23 % were positive for Giardia and 21 % for Cryptosporidium. An almost perfect agreement was found between IFAT and combined results of PCR, by Kappa composite proportion analysis. PCR positive samples were significantly more frequent in untreated raw water for C. parvum (p = 0.02). High mean of fecal coliforms, high pH values, and low mean of chlorine residuals were strongly correlated with PCR positivity for G. duodenalis assemblage B. High pH value was correlated with PCR positivity for C. parvum. Phylogenetic analysis of DNA sequences was possible, showing water and human clinical sequences for Toxoplasma within the same phylogenetic group for B1 repeated sequence. PCR assay is complementary to IFAT assay for monitoring of protozoa in raw and drinkable water, enabling species identification and to look for phylogenetic analysis in protozoa from human and environmental sources.

Prevalence of Cryptosporidium spp., Enterocytozoon bieneusi, Encephalitozoon spp. and Giardia intestinalis in Wild, Semi-Wild and Captive Orangutans (Pongo abelii and Pongo pygmaeus) on Sumatra and Borneo, Indonesia

PubMed Central

Mynářová, Anna; Foitová, Ivona; Kváč, Martin; Květoňová, Dana; Rost, Michael; Morrogh-Bernard, Helen; Nurcahyo, Wisnu; Nguyen, Cathleen; Supriyadi, Supriyadi; Sak, Bohumil

2016-01-01

Background Orangutans are critically endangered primarily due to loss and fragmentation of their natural habitat. This could bring them into closer contact with humans and increase the risk of zoonotic pathogen transmission. Aims To describe the prevalence and diversity of Cryptosporidium spp., microsporidia and Giardia intestinalis in orangutans at seven sites on Sumatra and Kalimantan, and to evaluate the impact of orangutans’ habituation and location on the occurrence of these zoonotic protists. Result The overall prevalence of parasites in 298 examined animals was 11.1%. The most prevalent microsporidia was Encephalitozoon cuniculi genotype II, found in 21 animals (7.0%). Enterocytozoon bieneusi genotype D (n = 5) and novel genotype Pongo 2 were detected only in six individuals (2.0%). To the best of our knowledge, this is the first report of these parasites in orangutans. Eight animals were positive for Cryptosporidium spp. (2.7%), including C. parvum (n = 2) and C. muris (n = 6). Giardia intestinalis assemblage B, subtype MB6, was identified in a single individual. While no significant differences between the different human contact level groups (p = 0.479–0.670) or between the different islands (p = 0.992) were reported in case of E. bieneusi or E. cuniculi, Cryptosporidium spp. was significantly less frequently detected in wild individuals (p < 2×10−16) and was significantly more prevalent in orangutans on Kalimantan than on Sumatra (p < 2×10−16). Conclusion Our results revealed that wild orangutans are significantly less frequently infected by Cryptosporidium spp. than captive and semi-wild animals. In addition, this parasite was more frequently detected at localities on Kalimantan. In contrast, we did not detect any significant difference in the prevalence of microsporidia between the studied groups of animals. The sources and transmission modes of infections were not determined, as this would require repeated sampling of individuals, examination of water sources, and sampling of humans and animals sharing the habitat with orangutans. PMID:27031241

Microbial pathogens in source and treated waters from drinking water treatment plants in the United States and implications for human health

EPA Science Inventory

An occurrence survey was conducted on selected pathogens in source and treated drinking water collected from 25 drinking water treatment plants (DWTPs) in the United States. Water samples were analyzed for the protozoa Giardia and Cryptosporidium (EPA Method 1623); the fungi Aspe...

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 observation of edible fillet contamination.

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 observation of edible fillet contamination. PMID:26213992

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. Copyright © 2016 Elsevier B.V. All rights reserved.

Subtype Analysis of Cryptosporidium Specimens from Sporadic Cases in Colorado, Idaho, New Mexico, and Iowa in 2007: Widespread Occurrence of One Cryptosporidium hominis Subtype and Case History of an Infection with the Cryptosporidium Horse Genotype▿

PubMed Central

Xiao, Lihua; Hlavsa, Michele C.; Yoder, Jonathan; Ewers, Christina; Dearen, Theresa; Yang, Wenli; Nett, Randall; Harris, Stephanie; Brend, Sarah M.; Harris, Meghan; Onischuk, Lisa; Valderrama, Amy L.; Cosgrove, Shaun; Xavier, Karen; Hall, Nancy; Romero, Sylvia; Young, Stephen; Johnston, Stephanie P.; Arrowood, Michael; Roy, Sharon; Beach, Michael J.

2009-01-01

Subtyping was conducted in late 2007 on 57 Cryptosporidium specimens from sporadic cases in Colorado, Idaho, New Mexico, and Iowa. One previously rare Cryptosporidium hominis subtype was indentified in 40 cases (70%) from all four states, and the Cryptosporidium horse genotype was identified in a pet shop employee with severe clinical symptoms. PMID:19587303

Evaluation of a magnetic microsphere antibody detection assay for the investigation of exposure to Cryptosporidium parvum and Cryptosporidium hominis

EPA Science Inventory

Anti-Cryptosporidium IgA and IgG are useful markers of exposure to Cryptosporidium in human populations, but detection in saliva may be difficult. To evaluate a magnetic microsphere assay for detection of anti-Cryptosporidium IgA and IgG in saliva, recombinant sporozoite gp15 (1...

Application of a salivary immunoassay in a prospective community study of waterborne infections.

PubMed

Egorov, Andrey I; Griffin, Shannon M; Ward, Honorine D; Reilly, Kevin; Fout, G Shay; Wade, Timothy J

2018-05-23

Quantifying sporadic waterborne infections in community settings can be challenging. Salivary antibody immunoassays are a promising non-invasive tool that can be used in prospective studies of common infections, especially those involving children. This study was conducted in a Massachusetts city, which uses a microbiologically contaminated river as its water source, during summer-early winter periods before and after construction of a new drinking water treatment plant. Monthly saliva samples (7480 samples from 1170 children and 816 adults) were analyzed for immunoglobulin G (IgG) responses to recombinant proteins of Cryptosporidium, one genogroup I (GI) and two GII noroviruses. Immunoconversion was defined as at least four-fold increase in specific antibody responses between two monthly samples with a post-conversion response above a flexible age-dependent cut-off. Episodes of gastroenteritis (diarrhea or vomiting or cramps) were associated with 3.2 (95% confidence limits 1.1; 9.5) adjusted odds ratio (aOR) of immunoconversion to Cryptosporidium; episodes of combined diarrhea and vomiting symptoms were associated with 3.5 (0.8; 15.0) and 4.6 (1.7; 12.6) aORs of an immunoconversion to GI and GII noroviruses, respectively. Swimming in natural water bodies or chlorinated pools was associated with 2.3 (0.4; 15.4) and 4.9 (1.6; 15.5) aORs of immunoconversion to Cryptosporidium, respectively. In a subset of study participants who did not use home water filters, consumption of at least some amount of non-boiled tap water reported in a monthly recall survey was associated with 11.1 (1.2; 100.0) and 0.6 (0.1; 2.5) aORs of immunoconversion to Cryptosporidium before and after the new water treatment plant construction, respectively. Among individuals who used home water filters, associations between non-boiled tap water consumption and Cryptosporidium immunoconversion were not significant before and after new plant construction with aORs of 0.8 (0.2; 3.3) and 0.3 (0.1; 1.6), respectively. The interaction effect of study phase and non-boiled tap water consumption on Cryptosporidium immunoconversions was statistically significant in the entire study population with aOR of 5.4 (1.1; 25.6). This was the first study that has used a salivary antibody immunoassay to demonstrate significant associations between gastrointestinal symptoms and Cryptosporidium and norovirus infections, and between water-related exposures and Cryptosporidium infections. Published by Elsevier Ltd.

A Modified EPA Method 1623 that Uses Tangential Flow Hollow-fiber Ultrafiltration and Heat Dissociation Steps to Detect Waterborne Cryptosporidium and Giardia spp.

PubMed Central

Rhodes, Eric R.; Villegas, Leah Fohl; Shaw, Nancy J.; Miller, Carrie; Villegas, Eric N.

2012-01-01

Cryptosporidium and Giardia species are two of the most prevalent protozoa that cause waterborne diarrheal disease outbreaks worldwide. To better characterize the prevalence of these pathogens, EPA Method 1623 was developed and used to monitor levels of these organisms in US drinking water supplies 12. The method has three main parts; the first is the sample concentration in which at least 10 L of raw surface water is filtered. The organisms and trapped debris are then eluted from the filter and centrifuged to further concentrate the sample. The second part of the method uses an immunomagnetic separation procedure where the concentrated water sample is applied to immunomagnetic beads that specifically bind to the Cryptosporidium oocysts and Giardia cysts allowing for specific removal of the parasites from the concentrated debris. These (oo)cysts are then detached from the magnetic beads by an acid dissociation procedure. The final part of the method is the immunofluorescence staining and enumeration where (oo)cysts are applied to a slide, stained, and enumerated by microscopy. Method 1623 has four listed sample concentration systems to capture Cryptosporidium oocysts and Giardia cysts in water: Envirochek filters (Pall Corporation, Ann Arbor, MI), Envirochek HV filters (Pall Corporation), Filta-Max filters (IDEXX, Westbrook, MA), or Continuous Flow Centrifugation (Haemonetics, Braintree, MA). However, Cryptosporidium and Giardia (oo)cyst recoveries have varied greatly depending on the source water matrix and filters used1,14. A new tangential flow hollow-fiber ultrafiltration (HFUF) system has recently been shown to be more efficient and more robust at recovering Cryptosporidium oocystsand Giardia cysts from various water matrices; moreover, it is less expensive than other capsule filter options and can concentrate multiple pathogens simultaneously1-3,5-8,10,11. In addition, previous studies by Hill and colleagues demonstrated that the HFUF significantly improved Cryptosporidium oocysts recoveries when directly compared with the Envirochek HV filters4. Additional modifications to the current methods have also been reported to improve method performance. Replacing the acid dissociation procedure with heat dissociation was shown to be more effective at separating Cryptosporidium from the magnetic beads in some matrices9,13 . This protocol describes a modified Method 1623 that uses the new HFUF filtration system with the heat dissociation step. The use of HFUF with this modified Method is a less expensive alternative to current EPA Method 1623 filtration options and provides more flexibility by allowing the concentration of multiple organisms. PMID:22805201

Widespread Occurrence of Zoonotic Cryptosporidium Species and Subtypes in Dairy Cattle from Northeast China: Public Health Concerns.

PubMed

Tao, Wei; Li, Yijing; Yang, Hang; Song, Mingxin; Lu, Yixin; Li, Wei

2018-02-01

Bovine cryptosporidiosis constitutes a threat to the livestock industry and public health worldwide. In the present study we investigated dairy cattle of all ages in northeast China for the prevalence and genetic traits of Cryptosporidium. Nested polymerase chain reaction of the small subunit rRNA gene was used to identify Cryptosporidium species or genotype. The parasite was detected in 130 of 537 (24.2%) animals sampled from the cities of Harbin (35.2%, 69/196) and Qiqihar (32.1%, 61/190). Cryptosporidium parvum (87/130) was identified as the dominant species by sequence analysis followed by Cryptosporidium bovis (28/130), Cryptosporidium ryanae (5/130), Cryptosporidium andersoni (2/130), Cryptosporidium suis-like genotype (2/130), and mixed C. ryanae/ C. bovis (1/130). Subtyping of C. parvum isolates was based on the DNA polymorphisms of the 60-kDa glycoprotein gene. Subtyping of the C. parvum isolates recognized subtypes IIdA15G1 (24/87) in Harbin and IIdA20G1 (48/87) in Qiqihar. A diversity of Cryptosporidium species/genotype and subtypes was identified in cattle from northeast China. Widespread occurrence of human-pathogenic Cryptosporidium species and subtypes is of public health significance. This is the first study reporting C. parvum subtype IIdA20G1 in China. The findings improve the epidemiological knowledge of bovine cryptosporidiosis in China, highlighting the importance of ongoing Cryptosporidium surveillance.

Detection and molecular characterization of Cryptosporidium spp. in captive canaries (Serinus canaria) using different diagnostic methods.

PubMed

Camargo, Vinícius da Silva; Santana, Bruna Nicoleti; Ferrari, Elis Domingos; Nakamura, Alex Akira; Nagata, Walter Bertequini; Nardi, Ana Rita Moraes; Meireles, Marcelo Vasconcelos

2018-01-01

This study used several diagnostic methods to examine the occurrence of and molecularly characterize Cryptosporidium spp. in captive canaries (Serinus canaria) in southern and southeastern Brazil. A total of 498 fecal samples were purified by centrifugal-flotation using Sheather's solution. Cryptosporidium spp. diagnosis was performed using three diagnostic methods: malachite green negative staining, nested PCR targeting the 18S rRNA gene, followed by sequencing the amplified fragments, and duplex real-time PCR targeting the 18S rRNA specific to detect Cryptosporidium galli and Cryptosporidium avian genotype III. The overall positivity for Cryptosporidium spp. (total samples positive in at least one protocol) from the microscopic analysis, nested PCR and duplex real-time PCR protocol results was 13.3% (66/498). The positivity rates were 2.0% (10/498) and 4.6% (23/498) for Cryptosporidium spp. by microscopy and nested PCR, respectively. Sequencing of 20 samples amplified by nested PCR identified C. galli (3.0%; 15/498), Cryptosporidium avian genotype I (0.8%; 4/498) and Cryptosporidium avium (0.2%; 1/498). Duplex real-time PCR revealed a positivity of 7.8% (39/498) for C. galli and 2.4% (12/498) for avian genotype III. Malachite green negative staining differed significantly from nested PCR in detecting Cryptosporidium spp. Duplex real-time PCR was more sensitive than nested PCR/sequencing for detecting gastric Cryptosporidium in canaries.

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.

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

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.

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. © 2011 Society for Risk Analysis.

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 risk management situations. Copyright © 2011 Elsevier Ltd. All rights reserved.

Occurrence, Molecular Characterization and Assessment of Zoonotic Risk of Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi in Pigs in Henan, Central China.

PubMed

Wang, Haiyan; Zhang, Yiqi; Wu, Yayun; Li, Junqiang; Qi, Meng; Li, Tingwen; Wang, Jianling; Wang, Rongjun; Zhang, Sumei; Jian, Fuchun; Ning, Changshen; Zhang, Longxian

2018-05-12

Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi are common gastrointestinal pathogens in humans and animals. Little is known about them and the range of species/assemblages/genotypes occurring in domestic pigs in China. Here, we present data on the occurrence and molecular diversity of these pathogens detected in the faeces from farms in Henan, central China. Of 897 fecal samples tested, 28 (3.1%), 15 (1.7%) and 408 (45.5%) samples were positive for Cryptosporidium, G. duodenalis and E. bieneusi, respectively. Cryptosporidium and G. duodenalis were most frequently detected in piglets, while E. bieneusi was markedly more prevalent in fattening pigs. Sequence analysis of SSU rRNA gene revealed that positive Cryptosporidium strains belonged to C. suis (n=18) and C. scrofarum (n=10). G. duodenalis assemblages E (n=9), assemblages A (n=3) and assemblages C (n=3) were characterized based on the sequence analysis of tpi gene. Thirteen E. bieneusi genotypes comprising four novel (pigHN-I to pigHN-IV) and nine known (EbpC, EbpA, pigEbITS5, LW1, H, CM8, G, CHG19, and CHS5) genotypes were identified by ITS sequence analysis of a large proportion (n=200) of E. bieneusi-positive samples. EbpC was the most frequent genotype, detected in 60 specimens. All 13 genotypes identified in this study clustered in zoonotic Group 1. The findings indicate that the presence of zoonotic species/assemblages/genotypes of these pathogens poses a threat to public health, suggesting that pigs in Henan province could be a significant source of human infection and water pollution. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Prevalence of cryptosporidium infection and characteristics of oocyst shedding in a breeding colony of leopard geckos (Eublepharis macularius).

PubMed

Deming, Clare; Greiner, Ellis; Uhl, Elizabeth W

2008-12-01

Cryptosporidiosis is an emerging problem in reptile medicine and has been associated with a wasting syndrome in leopard geckos (Eublepharis macularius). This study determined the prevalence of infection in a breeding colony of leopard geckos to be 9.8%. Two groups of 20 geckos, one that was fecal positive for oocysts of Cryptosporidium sp., and one, whose individuals were fecal negative at the inception of the study, were followed for 2 mo. Fecal samples were tested for oocysts every 2 wk, body weights were measured, and a body condition score was assigned for each gecko. Selected geckos from these two groups were euthanized and necropsied. There were statistically significant differences (P < 0.05) between the two groups for mean body weight, mean body condition score, and prevalence of infection. Cryptosporidium sp. infection is endemic in this breeding colony, and there were a large number of geckos with a subclinical or carrier state of infection. These animals continued to be infected with Cryptosporidium sp. but gained weight and remained in good body condition. Only one gecko in the entire group of 40 was confirmed to be negative for oocysts or developmental stages by repeated fecal exams and histopathology. An additional 37 severely emaciated geckos from the breeding colony were euthanized, and all were positive for Cryptosporidium sp. on histopathologic examination of the gastrointestinal tract. The results of this study indicate that although some animals can recover from a clinical infection, if a gecko is severely wasted, it should be euthanized because of the poor prognosis and possible source of infection to other geckos.

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.

OCCURRENCE AND EXPOSURE ASSESSMENT FOR THE ...

EPA Pesticide Factsheets

Describes the occurrence of Cryptosporidium and other pathogens in the raw and finished water of public water systems (PWS) based on modeling of source water survey data. Analysis of microbial occurrence data to support LT2ESWTR microbial risk assessment

Pathogen Loading From Canada Geese Faeces in Freshwater: Potential Risks to Human Health Through Recreational Water Exposure.

PubMed

Gorham, T J; Lee, J

2016-05-01

Canada geese (Branta canadensis) faeces have been shown to contain pathogenic protozoa and bacteria in numerous studies over the past 15 years. Further, increases in both the Canada geese populations and their ideal habitat requirements in the United States (US) translate to a greater presence of these human pathogens in public areas, such as recreational freshwater beaches. Combining these factors, the potential health risk posed by Canada geese faeces at freshwater beaches presents an emerging public health issue that warrants further study. Here, literature concerning human pathogens in Canada geese faeces is reviewed and the potential impacts these pathogens may have on human health are discussed. Pathogens of potential concern include Campylobacter jejuni, Salmonella Typhimurium, Listeria monocytogenes, Helicobacter canadensis, Arcobacter spp., Enterohemorragic Escherichia coli pathogenic strains, Chlamydia psitacci, Cryptosporidium parvum and Giardia lamblia. Scenarios presenting potential exposure to pathogens eluted from faeces include bathers swimming in lakes, children playing with wet and dry sand impacted by geese droppings and other common recreational activities associated with public beaches. Recent recreational water-associated disease outbreaks in the US support the plausibility for some of these pathogens, including Cryptosporidium spp. and C. jejuni, to cause human illness in this setting. In view of these findings and the uncertainties associated with the real health risk posed by Canada geese faecal pathogens to users of freshwater lakes, it is recommended that beach managers use microbial source tracking and conduct a quantitative microbial risk assessment to analyse the local impact of Canada geese on microbial water quality during their decision-making process in beach and watershed management. © 2015 Blackwell Verlag GmbH.

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. Copyright © 2016 Elsevier B.V. All rights reserved.

Occurrence of Giardia intestinalis and Cryptosporidium sp. in wastewater samples from São Paulo State, Brazil, and Lima, Peru.

PubMed

Ulloa-Stanojlović, Francisco Miroslav; Aguiar, Bruna; Jara, Luis M; Sato, Maria Inês Zanoli; Guerrero, Juana Arzola; Hachich, Elayse; Matté, Glavur Rogério; Dropa, Milena; Matté, Maria Helena; de Araújo, Ronalda Silva

2016-11-01

The objectives of the study were to detect and genotype Cryptosporidium spp. and Giardia intestinalis in wastewater samples obtained from five cities with high transit of people in the State of São Paulo, Brazil, and at the entrance of a Wastewater Treatment Plant (WWTP) in Lima, Peru. Samples were collected and concentrated by centrifugation. The genomic DNA was extracted for molecular characterization by nested PCR for Cryptosporidium and double nested PCR for Giardia, followed by sequencing and phylogenetic analysis. G. intestinalis was found in 63.6 % of the samples, and the human assemblages A and B were identified. Cryptosporidium sp. was found in 36.4 % of the samples, and the species were corresponding to Cryptosporidium hominis, Cryptosporidium cuniculus, and Cryptosporidium muris. Results revealed the presence of human pathogenic Cryptosporidium species and G. intestinalis human pathogenic assemblages. Molecular tools highlight the importance to map the genetic diversity of these parasites, as well as to detect their epidemiological circulation pathway in the environment.

Molecular Characterization of Cryptosporidium Species and Giardia duodenalis from Symptomatic Cambodian Children.

PubMed

Moore, Catrin E; Elwin, Kristin; Phot, Nget; Seng, Chanthou; Mao, Saroeun; Suy, Kuong; Kumar, Varun; Nader, Johanna; Bousfield, Rachel; Perera, Sanuki; Bailey, J Wendi; Beeching, Nicholas J; Day, Nicholas P J; Parry, Christopher M; Chalmers, Rachel M

2016-07-01

In a prospective study, 498 single faecal samples from children aged under 16 years attending an outpatient clinic in the Angkor Hospital for Children, northwest Cambodia, were examined for Cryptosporidium oocysts and Giardia cysts using microscopy and molecular assays. Cryptosporidium oocysts were detected in 2.2% (11/498) of samples using microscopy and in 7.7% (38/498) with molecular tests. Giardia duodenalis cysts were detected in 18.9% (94/498) by microscopy and 27.7% (138/498) by molecular tests; 82% of the positive samples (by either method) were from children aged 1-10 years. Cryptosporidium hominis was the most common species of Cryptosporidium, detected in 13 (34.2%) samples, followed by Cryptosporidium meleagridis in 9 (23.7%), Cryptosporidium parvum in 8 (21.1%), Cryptosporidium canis in 5 (13.2%), and Cryptosporidium suis and Cryptosporidium ubiquitum in one sample each. Cryptosporidium hominis and C. parvum positive samples were subtyped by sequencing the GP60 gene: C. hominis IaA16R6 and C. parvum IIeA7G1 were the most abundant subtypes. Giardia duodenalis was typed using a multiplex real-time PCR targeting assemblages A and B. Assemblage B (106; 76.8% of all Giardia positive samples) was most common followed by A (12.3%) and mixed infections (5.1%). Risk factors associated with Cryptosporidium were malnutrition (AOR 9.63, 95% CI 1.67-55.46), chronic medical diagnoses (AOR 4.51, 95% CI 1.79-11.34) and the presence of birds in the household (AOR 2.99, 95% CI 1.16-7.73); specifically C. hominis (p = 0.03) and C. meleagridis (p<0.001) were associated with the presence of birds. The use of soap was protective against Giardia infection (OR 0.74, 95% CI 0.58-0.95). This is the first report to describe the different Cryptosporidium species and subtypes and Giardia duodenalis assemblages in Cambodian children. The variety of Cryptosporidium species detected indicates both anthroponotic and zoonotic transmission in this population. Interventions to improve sanitation, increase hand washing after defecation and before preparing food and promote drinking boiled water may reduce the burden of these two parasites.

40 CFR 141.721 - Reporting requirements.

Code of Federal Regulations, 2012 CFR

2012-07-01

... Section 141.721 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Reporting and... § 141.702 and source water monitoring results under § 141.706 unless they notify the State that they...

40 CFR 141.721 - Reporting requirements.

Code of Federal Regulations, 2014 CFR

2014-07-01

... Section 141.721 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Reporting and... § 141.702 and source water monitoring results under § 141.706 unless they notify the State that they...

40 CFR 141.721 - Reporting requirements.

Code of Federal Regulations, 2011 CFR

2011-07-01

... Section 141.721 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Reporting and... § 141.702 and source water monitoring results under § 141.706 unless they notify the State that they...

40 CFR 141.721 - Reporting requirements.

Code of Federal Regulations, 2013 CFR

2013-07-01

... Section 141.721 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Reporting and... § 141.702 and source water monitoring results under § 141.706 unless they notify the State that they...

40 CFR 141.711 - Filtered system additional Cryptosporidium treatment requirements.

Code of Federal Regulations, 2013 CFR

2013-07-01

... AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced...(including softening) Direct filtration Slow sand or diatomaceous earth filtration Alternative filtration... survey or an equivalent source water assessment that after a system completed the monitoring conducted...

40 CFR 141.711 - Filtered system additional Cryptosporidium treatment requirements.

Code of Federal Regulations, 2012 CFR

2012-07-01

... AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced...(including softening) Direct filtration Slow sand or diatomaceous earth filtration Alternative filtration... survey or an equivalent source water assessment that after a system completed the monitoring conducted...

40 CFR 141.711 - Filtered system additional Cryptosporidium treatment requirements.

Code of Federal Regulations, 2014 CFR

2014-07-01

... AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced...(including softening) Direct filtration Slow sand or diatomaceous earth filtration Alternative filtration... survey or an equivalent source water assessment that after a system completed the monitoring conducted...

Evaluation of ColorPAC Giardia/Cryptosporidium Rapid Assay and ProSpecT Giardia/Cryptosporidium Microplate Assay for Detection of Giardia and Cryptosporidium in Fecal Specimens

PubMed Central

Katanik, M. T.; Schneider, S. K.; Rosenblatt, J. E.; Hall, G. S.; Procop, G. W.

2001-01-01

Detection of Giardia and Cryptosporidium in clinical stool specimens using the ColorPAC and ProSpecT enzyme immunoassays revealed 98.7% agreement for Giardia detection and 98.1% agreement for Cryptosporidium detection. Sensitivities were uniformly 100%. The specificities of the ColorPAC immunoassay for Giardia and Cryptosporidium detection were 100 and 99.5%, respectively, and those for the ProSpecT assay were 98.4 and 98.6%, respectively. The false-positive reactions with the ProSpecT assay occurred with specimens that were grossly bloody. PMID:11724874

Cloaca prolapse and cystitis in green iguana (Iguana iguana) caused by a novel Cryptosporidium species.

PubMed

Kik, Marja J L; van Asten, Alphons J A M; Lenstra, Johannes A; Kirpensteijn, Jolle

2011-01-10

Cryptosporidium infection was associated with colitis and cystitis in 2 green iguanas (Iguana iguana). The disease was characterized by a chronic clinical course of cloacal prolapses and cystitis. Histological examination of the gut and urinary bladder showed numerous Cryptosporidium developmental stages on the surface of the epithelium with mixed inflammatory response in the lamina propria. Cryptosporidium oocysts were visualised in a cytological preparation of the faeces. Based on the small subunit ribosomal RNA gene the cryptosporidia were characterized as belonging to the intestinal cryptosporidial lineage, but not to Cryptosporidium saurophilum or Cryptosporidium serpentis species. Copyright © 2010 Elsevier B.V. All rights reserved.

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...

INVESTIGATION ON CRYPTOSPORIDIUM INFECTIONS IN WILD ANIMALS IN A ZOO IN ANHUI PROVINCE.

PubMed

Gu, Youfang; Wang, Xiaolong; Zhou, Cefan; Li, Peiying; Xu, Qianming; Zhao, Changcheng; Liu, Wei; Xu, Wenlong

2016-09-01

To assess Cryptosporidium infections among wild animals in a zoo located in Anhui province, we conducted an investigation on the fecal samples collected from 44 primates, 41 herbivores, 44 carnivores and omnivores, and 103 birds in the zoo with the use of Sheather's sugar flotation technique and modified acid-fast staining. Cryptosporidium oocysts were detected in the fecal samples from six primates, two herbivores, four carnivores and omnivores, and seven birds by using Sheather's sugar flotation technique; the prevalence of Cryptosporidium infection in primates, herbivores, carnivores and omnivores and birds was 13.64, 4.88, 9.09, and 6.80%, respectively. Modified acid-fast staining detected the presence of Cryptosporidium oocysts in the fecal samples of one primate, three herbivores, 0 carnivores and omnivores, and one bird, and the prevalence of Cryptosporidium infection in primates, herbivores, carnivores and omnivores and birds was 2.27, 7.32, 0.00, and 0.97%, respectively. Polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) and phylogenetic analysis with the use of the neighbor-joining (NJ) method based on the aligned partial small-subunit (SSU) rRNA gene sequences showed that the protozoan pathogen isolated from primates was Cryptosporidium hominis and the pathogen isolated from camels ( Camelus dromedarius ) was Cryptosporidium andersoni. Subtyping the Cryptosporidium hominis by 60-kDa glycoprotein (GP60) gene phylogenetic analysis showed the Cryptosporidium hominis belongs to the subtype IdA and IbA.

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...

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.

Prevalence of Cryptosporidium-like infection in one-humped camels (Camelus dromedarius) of northwestern Iran

PubMed Central

Yakhchali, M.; Moradi, T.

2012-01-01

Cryptosporidium is a ubiquitous enteropathogen protozoan infection affecting livestock worldwide. The present study was carried out to determine the prevalence of Cryptosporidium infection in different age groups of dromedary camels in northwestern Iran from November 2009 to July 2010. A total number of 170 fecal samples were collected and examined using modified Ziehl-Neelsen (MZN) staining under light microscope. Examination of stained fecal smears revealed that 17 camels (10%) were positive for Cryptosporidium-like. The prevalence of Cryptosporidium-like was significantly higher in camel calves (< 1 years old) (20%) than other age groups, in which the diarrhoeic calves had the prevalence of 16%. In adult camels the prevalence was 6.5%. There was no significant difference in the prevalence of Cryptosporidium-like between male and female camels. It is concluded that Cryptosporidium infection is a problem in camel husbandry and could be of public health concern in the region. PMID:22314242

Prevalence of Cryptosporidium-like infection in one-humped camels (Camelus dromedarius) of northwestern Iran.

PubMed

Yakhchali, M; Moradi, T

2012-02-01

Cryptosporidium is a ubiquitous enteropathogen protozoan infection affecting livestock worldwide. The present study was carried out to determine the prevalence of Cryptosporidium infection in different age groups of dromedary camels in northwestern Iran from November 2009 to July 2010. A total number of 170 fecal samples were collected and examined using modified Ziehl-Neelsen (MZN) staining under light microscope. Examination of stained fecal smears revealed that 17 camels (10%) were positive for Cryptosporidium-like. The prevalence of Cryptosporidium-like was significantly higher in camel calves (< 1 years old) (20%) than other age groups, in which the diarrhoeic calves had the prevalence of 16%. In adult camels the prevalence was 6.5%. There was no significant difference in the prevalence of Cryptosporidium-like between male and female camels. It is concluded that Cryptosporidium infection is a problem in camel husbandry and could be of public health concern in the region.

The first study of molecular prevalence and species characterization of Cryptosporidium in free-range chicken (Gallus gallus domesticus) from Brazil.

PubMed

Ewald, Maria Paula de Carvalho; Martins, Felippe Danyel Cardoso; Caldart, Eloiza Teles; Vieira, Fernando Emmanuel Gonçalves; Yamamura, Milton Hissashi; Sasse, João Pedro; Barros, Luiz Daniel de; Freire, Roberta Lemos; Navarro, Italmar Teodorico; Garcia, João Luis

2017-01-01

Rearing free-range chicken is based on grazing feeding patterns, and these animals could be potential environmental contaminants of Cryptosporidium oocysts for humans and other animals. Therefore, the present study aimed to evaluate the molecular prevalence of Cryptosporidium spp. in free-range chickens from Brazil. A total of 351 fecal samples from chickens were examined from 20 farms. For detection of Cryptosporidium spp., 18S rRNA gene fragments were amplified using a nested PCR reaction. Positive samples were sent for sequencing. The overall prevalence of Cryptosporidium was 25.6% (95% CI = 21.2% - 30.6%). Sequencing of the amplified fragments allowed for the identification of three species: C. meleagridis in 57 (62.6%), C. baileyi in 15 (16.4%), C. parvum in 3 (3.2%) samples, and a new Cryptosporidium genotype (C. genotype BrPR1) in 3 (3.2%) samples. Cryptosporidium genotype BrPR1 has not yet been classified as a species, and its host spectrum is not known. Cryptosporidium, including zoonotic species, exists at a high prevalence in free-range chickens within the region studied.

Cryptosporidium and Giardia in marine-foraging river otters (Lontra canadensis) from the Puget Sound Georgia Basin ecosystem.

PubMed

Gaydos, J K; Miller, W A; Gilardi, K V K; Melli, A; Schwantje, H; Engelstoft, C; Fritz, H; Conrad, P A

2007-02-01

Species of Cryptosporidium and Giardia can infect humans and wildlife and have the potential to be transmitted between these 2 groups; yet, very little is known about these protozoans in marine wildlife. Feces of river otters (Lontra canadensis), a common marine wildlife species in the Puget Sound Georgia Basin, were examined for species of Cryptosporidium and Giardia to determine their role in the epidemiology of these pathogens. Using ZnSO4 flotation and immunomagnetic separation, followed by direct immunofluorescent antibody detection (IMS/DFA), we identified Cryptosporidium sp. oocysts in 9 fecal samples from 6 locations and Giardia sp. cysts in 11 fecal samples from 7 locations. The putative risk factors of proximate human population and degree of anthropogenic shoreline modification were not associated with the detection of Cryptosporidium or Giardia spp. in river otter feces. Amplification of DNA from the IMS/DFA slide scrapings was successful for 1 sample containing > 500 Cryptosporidium sp. oocysts. Sequences from the Cryptosporidium 18S rRNA and the COWP loci were most similar to the ferret Cryptosporidium sp. genotype. River otters could serve as reservoirs for Cryptosporidium and Giardia species in marine ecosystems. More work is needed to better understand the zoonotic potential of the genotypes they carry as well as their implications for river otter health.

Cryptosporidium oocysts and giardia cysts on salad products irrigated with contaminated water.

PubMed

Amorós, Inmaculada; Alonso, José L; Cuesta, Gonzalo

2010-06-01

A field study in Valencia, Spain, was done to determine the occurrence of Giardia and Cryptosporidium on salad products that are frequently eaten raw, such as lettuces and Chinese cabbage, and in irrigation waters. Four water samples were taken weekly 1 month before harvesting the vegetables. All water samples were analyzed using techniques included in the U.S. Environmental Protection Agency Method 1623. Standard methods for detecting protozoan parasites on salad vegetables are not available. Published techniques for the isolation of parasites from vegetables generally have low and variable recovery efficiencies. In this study, vegetables were analyzed using a recently reported method for detection of Cryptosporidium oocysts and Giardia cysts on salad products. The waters tested were positive for both Cryptosporidium and Giardia. Of 19 salad products studied, we observed Cryptosporidium in 12 samples and Giardia in 10 samples. Recoveries of the Texas Red-stained Cryptosporidium and Giardia, which were used as internal controls, were 24.5% +/- 3.5% for Cryptosporidium and 16.7% +/- 8.1% for Giardia (n = 8). This study provides data on the occurrence of Cryptosporidium and Giardia in salad products in Spain. The method was useful in the detection of Cryptosporidium oocysts and Giardia cysts on the vegetables tested, and it provides a useful analytical tool for occurrence monitoring.

Prevalence and molecular characterization of Cryptosporidium in giant panda (Ailuropoda melanoleuca) in Sichuan province, China.

PubMed

Wang, Tao; Chen, Zuqin; Xie, Yue; Hou, Rong; Wu, Qidun; Gu, Xiaobing; Lai, Weiming; Peng, Xuerong; Yang, Guangyou

2015-06-25

Cryptosporidium spp. have been extensively reported to cause significant diarrheal disease in humans and domestic animals. On the contrary, little information is available on the prevalence and characterization of Cryptosporidium in wild animals in China, especially in giant pandas. The aim of the present study was to detect Cryptosporidium infections and identify Cryptosporidium species at the molecular level in both captive and wild giant pandas in Sichuan province, China. Using a PCR approach, we amplified and sequenced the 18S rRNA gene from 322 giant pandas fecal samples (122 from 122 captive individuals and 200 collected from four habitats) in Sichuan province, China. The Cryptosporidium species/genotypes were identified via a BLAST comparison against published Cryptosporidium sequences available in GenBank followed by phylogenetic analysis. The results revealed that both captive and wild giant pandas were infected with a single Cryptosporidium species, C. andersoni, at a prevalence of 15.6% (19/122) and 0.5% (1/200) in captive and wild giant pandas, respectively. The present study revealed the existence of C. andersoni in both captive and wild giant panda fecal samples for the first time, and also provided useful fundamental data for further research on the molecular epidemiology and control of Cryptosporidium infection in giant pandas.

Assessment of microscopic and molecular tools for the diagnosis and follow-up of cryptosporidiosis in patients at risk.

PubMed

Le Govic, Y; Guyot, K; Certad, G; Deschildre, A; Novo, R; Mary, C; Sendid, B; Viscogliosi, E; Favennec, L; Dei-Cas, E; Fréalle, E; Dutoit, E

2016-01-01

Cryptosporidiosis is an important though underreported public health concern. Molecular tools might be helpful in improving its diagnosis. In this study, ZR Fecal DNA MiniPrep™ Kit (ZR) and NucliSens® easyMAG® (EM) were compared using four Cryptosporidium-seeded feces and 29 Cryptosporidium-positive stools. Thereafter, ZR was selected for prospective evaluation of Cryptosporidium detection by 18S rDNA and LAXER quantitative PCR (qPCR) in 69 stools from 56 patients after Cryptosporidium detection by glycerin, modified Ziehl-Neelsen (ZN) and auramine-phenol (AP) stainings. The combination of any of the two extraction methods with 18S qPCR yielded adequate detection of Cryptosporidium in seeded stools, but the ZR kit showed the best performance. All 29 Cryptosporidium-positive samples were positive with 18S qPCR, after both ZR and EM extraction. However, false-negative results were found with LAXER qPCR or nested PCR. Cryptosporidiosis was diagnosed in 7/56 patients. All the microscopic methods enabled the initial diagnosis, but Cryptosporidium was detected in 12, 13, and 14 samples from these seven patients after glycerin, ZN, and AP staining respectively. Among these samples, 14 and 12 were positive with 18S and LAXER qPCR respectively. In two patients, Cryptosporidium DNA loads were found to be correlated with clinical evolution. Although little known, glycerin is a sensitive method for the initial detection of Cryptosporidium. When combined with 18S qPCR, ZR extraction, which had not been evaluated so far for Cryptosporidium, was an accurate tool for detecting Cryptosporidium and estimating the oocyst shedding in the course of infection.

Assessment of Zoonotic Transmission of Giardia and Cryptosporidium between Cattle and Humans in Rural Villages in Bangladesh

PubMed Central

Ehsan, Amimul M.; Geurden, Thomas; Casaert, Stijn; Parvin, Sonia M.; Islam, Taohidul M.; Ahmed, Uddin M.; Levecke, Bruno; Vercruysse, Jozef; Claerebout, Edwin

2015-01-01

Giardia and Cryptosporidium are important causes of diarrhoea in Bangladesh. The high prevalence of both parasites in humans and cattle in rural Bangladesh and the common use of water ponds by village inhabitants and their animals suggest a potential for zoonotic transmission. Direct transmission of Giardia and Cryptosporidium between cattle and their handlers and indirect transmission through water ponds was investigated. Faecal/stool samples were collected from 623 calves and 125 calf handlers in a cross-sectional survey. In two villages, water samples were collected monthly from water ponds and faecal/stool samples were collected monthly from inhabitants and their cattle. Giardia cysts and Cryptosporidium oocysts were detected in water samples and in faecal/stool samples and positive samples were genotyped, to determine their human or animal origin. The prevalence of Giardia and Cryptosporidium in calves was 22% and 5% respectively. In calf handlers, the prevalence of Giardia and Cryptosporidium was 11.2% and 3.2% respectively. Both in the cross-sectional survey and in the longitudinal study in the villages, G. duodenalis assemblage E was most prevalent in calves, while in humans assemblage AII, BIII and BIV were found. In cattle, Cryptosporidium parvum, C. bovis and C. andersoni were identified, but no Cryptosporidium sequences were obtained from humans. Giardia and Cryptosporidium were detected in 14/24 and 12/24 water samples respectively. G. duodenalis assemblage E and BIV (-like), as well as C. andersoni and C. hominis were identified. Although the presence of Giardia and Cryptosporidium in both water ponds suggests that water-borne transmission of Giardia and Cryptosporidium is possible, the genotyping results indicate that there is no significant direct or indirect (water-borne) transmission of Giardia between cattle and people in this area of rural Bangladesh. No conclusions could be drawn for Cryptosporidium, because of the low number of sequences that were obtained from human and water samples. PMID:25695662

Assessment of zoonotic transmission of Giardia and Cryptosporidium between cattle and humans in rural villages in Bangladesh.

PubMed

Ehsan, Amimul M; Geurden, Thomas; Casaert, Stijn; Parvin, Sonia M; Islam, Taohidul M; Ahmed, Uddin M; Levecke, Bruno; Vercruysse, Jozef; Claerebout, Edwin

2015-01-01

Giardia and Cryptosporidium are important causes of diarrhoea in Bangladesh. The high prevalence of both parasites in humans and cattle in rural Bangladesh and the common use of water ponds by village inhabitants and their animals suggest a potential for zoonotic transmission. Direct transmission of Giardia and Cryptosporidium between cattle and their handlers and indirect transmission through water ponds was investigated. Faecal/stool samples were collected from 623 calves and 125 calf handlers in a cross-sectional survey. In two villages, water samples were collected monthly from water ponds and faecal/stool samples were collected monthly from inhabitants and their cattle. Giardia cysts and Cryptosporidium oocysts were detected in water samples and in faecal/stool samples and positive samples were genotyped, to determine their human or animal origin. The prevalence of Giardia and Cryptosporidium in calves was 22% and 5% respectively. In calf handlers, the prevalence of Giardia and Cryptosporidium was 11.2% and 3.2% respectively. Both in the cross-sectional survey and in the longitudinal study in the villages, G. duodenalis assemblage E was most prevalent in calves, while in humans assemblage AII, BIII and BIV were found. In cattle, Cryptosporidium parvum, C. bovis and C. andersoni were identified, but no Cryptosporidium sequences were obtained from humans. Giardia and Cryptosporidium were detected in 14/24 and 12/24 water samples respectively. G. duodenalis assemblage E and BIV (-like), as well as C. andersoni and C. hominis were identified. Although the presence of Giardia and Cryptosporidium in both water ponds suggests that water-borne transmission of Giardia and Cryptosporidium is possible, the genotyping results indicate that there is no significant direct or indirect (water-borne) transmission of Giardia between cattle and people in this area of rural Bangladesh. No conclusions could be drawn for Cryptosporidium, because of the low number of sequences that were obtained from human and water samples.

Molecular-based investigation of Cryptosporidium and Giardia from animals in water catchments in southeastern Australia.

PubMed

Nolan, Matthew J; Jex, Aaron R; Koehler, Anson V; Haydon, Shane R; Stevens, Melita A; Gasser, Robin B

2013-04-01

There has been no large-scale systematic molecular epidemiological investigation of the waterborne protozoans, Cryptosporidium or Giardia, in southeastern Australia. Here, we explored, for the first time, the genetic composition of these genera in faecal samples from animals in nine Melbourne Water reservoir areas, collected over a period of two-years. We employed PCR-based single-strand conformation polymorphism (SSCP) and phylogenetic analyses of loci (pSSU and pgp60) in the small subunit (SSU) of ribosomal RNA and 60-kDa glycoprotein (gp60) genes to detect and characterise Cryptosporidium, and another locus (ptpi) in the triose-phosphate isomerase (tpi) gene to identify and characterise Giardia. Cryptosporidium was detected in 2.8% of the 2009 samples examined; the analysis of all amplicons defined 14 distinct sequence types for each of pSSU and pgp60, representing Cryptosporidium hominis (genotype Ib - subgenotype IbA10G2R2), Cryptosporidium parvum (genotype IIa - subgenotypes IIaA15G2R1, IIaA19G2R1, IIaA19G3R1, IIaA19G4R1, IIaA20G3R1, IIaA20G4R1, IIaA20G3R2 and IIaA21G3R1), Cryptosporidium cuniculus (genotype Vb - subgenotypes VbA22R4, VbA23R3, VbA24R3, VbA25R4 and VbA26R4), and Cryptosporidium canis, Cryptosporidium fayeri, Cryptosporidium macropodum and Cryptosporidium ubiquitum as well as six new pSSU sequence types. In addition, Giardia was identified in 3.4% of the samples; all 28 distinct ptpi sequence types defined were linked to assemblage A of Giardia duodenalis. Of all 56 sequence types characterised, eight and one have been recorded previously in Cryptosporidium and Giardia, respectively, from humans. In contrast, nothing is known about the zoonotic potential of 35 new genotypes of Cryptosporidium and Giardia recorded here for the first time. Future work aims to focus on estimating the prevalence of Cryptosporidium and Giardia genotypes in humans and a wide range of animals in Victoria and elsewhere in Australia. (Nucleotide sequences reported in this paper are available in the GenBank database under accession nos. KC282952-KC283005). Copyright © 2013. Published by Elsevier Ltd.

The Burden of Cryptosporidium Diarrheal Disease among Children < 24 Months of Age in Moderate/High Mortality Regions of Sub-Saharan Africa and South Asia, Utilizing Data from the Global Enteric Multicenter Study (GEMS).

PubMed

Sow, Samba O; Muhsen, Khitam; Nasrin, Dilruba; Blackwelder, William C; Wu, Yukun; Farag, Tamer H; Panchalingam, Sandra; Sur, Dipika; Zaidi, Anita K M; Faruque, Abu S G; Saha, Debasish; Adegbola, Richard; Alonso, Pedro L; Breiman, Robert F; Bassat, Quique; Tamboura, Boubou; Sanogo, Doh; Onwuchekwa, Uma; Manna, Byomkesh; Ramamurthy, Thandavarayan; Kanungo, Suman; Ahmed, Shahnawaz; Qureshi, Shahida; Quadri, Farheen; Hossain, Anowar; Das, Sumon K; Antonio, Martin; Hossain, M Jahangir; Mandomando, Inacio; Nhampossa, Tacilta; Acácio, Sozinho; Omore, Richard; Oundo, Joseph O; Ochieng, John B; Mintz, Eric D; O'Reilly, Ciara E; Berkeley, Lynette Y; Livio, Sofie; Tennant, Sharon M; Sommerfelt, Halvor; Nataro, James P; Ziv-Baran, Tomer; Robins-Browne, Roy M; Mishcherkin, Vladimir; Zhang, Jixian; Liu, Jie; Houpt, Eric R; Kotloff, Karen L; Levine, Myron M

2016-05-01

The importance of Cryptosporidium as a pediatric enteropathogen in developing countries is recognized. Data from the Global Enteric Multicenter Study (GEMS), a 3-year, 7-site, case-control study of moderate-to-severe diarrhea (MSD) and GEMS-1A (1-year study of MSD and less-severe diarrhea [LSD]) were analyzed. Stools from 12,110 MSD and 3,174 LSD cases among children aged <60 months and from 21,527 randomly-selected controls matched by age, sex and community were immunoassay-tested for Cryptosporidium. Species of a subset of Cryptosporidium-positive specimens were identified by PCR; GP60 sequencing identified anthroponotic C. parvum. Combined annual Cryptosporidium-attributable diarrhea incidences among children aged <24 months for African and Asian GEMS sites were extrapolated to sub-Saharan Africa and South Asian regions to estimate region-wide MSD and LSD burdens. Attributable and excess mortality due to Cryptosporidium diarrhea were estimated. Cryptosporidium was significantly associated with MSD and LSD below age 24 months. Among Cryptosporidium-positive MSD cases, C. hominis was detected in 77.8% (95% CI, 73.0%-81.9%) and C. parvum in 9.9% (95% CI, 7.1%-13.6%); 92% of C. parvum tested were anthroponotic genotypes. Annual Cryptosporidium-attributable MSD incidence was 3.48 (95% CI, 2.27-4.67) and 3.18 (95% CI, 1.85-4.52) per 100 child-years in African and Asian infants, respectively, and 1.41 (95% CI, 0.73-2.08) and 1.36 (95% CI, 0.66-2.05) per 100 child-years in toddlers. Corresponding Cryptosporidium-attributable LSD incidences per 100 child-years were 2.52 (95% CI, 0.33-5.01) and 4.88 (95% CI, 0.82-8.92) in infants and 4.04 (95% CI, 0.56-7.51) and 4.71 (95% CI, 0.24-9.18) in toddlers. We estimate 2.9 and 4.7 million Cryptosporidium-attributable cases annually in children aged <24 months in the sub-Saharan Africa and India/Pakistan/Bangladesh/Nepal/Afghanistan regions, respectively, and ~202,000 Cryptosporidium-attributable deaths (regions combined). ~59,000 excess deaths occurred among Cryptosporidium-attributable diarrhea cases over expected if cases had been Cryptosporidium-negative. The enormous African/Asian Cryptosporidium disease burden warrants investments to develop vaccines, diagnostics and therapies.

The Burden of Cryptosporidium Diarrheal Disease among Children < 24 Months of Age in Moderate/High Mortality Regions of Sub-Saharan Africa and South Asia, Utilizing Data from the Global Enteric Multicenter Study (GEMS)

PubMed Central

Nasrin, Dilruba; Blackwelder, William C.; Wu, Yukun; Farag, Tamer H.; Panchalingam, Sandra; Sur, Dipika; Zaidi, Anita K. M.; Faruque, Abu S. G.; Saha, Debasish; Adegbola, Richard; Alonso, Pedro L.; Breiman, Robert F.; Bassat, Quique; Tamboura, Boubou; Sanogo, Doh; Onwuchekwa, Uma; Manna, Byomkesh; Ramamurthy, Thandavarayan; Kanungo, Suman; Ahmed, Shahnawaz; Qureshi, Shahida; Quadri, Farheen; Hossain, Anowar; Das, Sumon K.; Antonio, Martin; Hossain, M. Jahangir; Mandomando, Inacio; Nhampossa, Tacilta; Acácio, Sozinho; Omore, Richard; Oundo, Joseph O.; Ochieng, John B.; Mintz, Eric D.; O’Reilly, Ciara E.; Berkeley, Lynette Y.; Livio, Sofie; Tennant, Sharon M.; Sommerfelt, Halvor; Nataro, James P.; Ziv-Baran, Tomer; Robins-Browne, Roy M.; Mishcherkin, Vladimir; Zhang, Jixian; Liu, Jie; Houpt, Eric R.; Kotloff, Karen L.; Levine, Myron M.

2016-01-01

Background The importance of Cryptosporidium as a pediatric enteropathogen in developing countries is recognized. Methods Data from the Global Enteric Multicenter Study (GEMS), a 3-year, 7-site, case-control study of moderate-to-severe diarrhea (MSD) and GEMS-1A (1-year study of MSD and less-severe diarrhea [LSD]) were analyzed. Stools from 12,110 MSD and 3,174 LSD cases among children aged <60 months and from 21,527 randomly-selected controls matched by age, sex and community were immunoassay-tested for Cryptosporidium. Species of a subset of Cryptosporidium-positive specimens were identified by PCR; GP60 sequencing identified anthroponotic C. parvum. Combined annual Cryptosporidium-attributable diarrhea incidences among children aged <24 months for African and Asian GEMS sites were extrapolated to sub-Saharan Africa and South Asian regions to estimate region-wide MSD and LSD burdens. Attributable and excess mortality due to Cryptosporidium diarrhea were estimated. Findings Cryptosporidium was significantly associated with MSD and LSD below age 24 months. Among Cryptosporidium-positive MSD cases, C. hominis was detected in 77.8% (95% CI, 73.0%-81.9%) and C. parvum in 9.9% (95% CI, 7.1%-13.6%); 92% of C. parvum tested were anthroponotic genotypes. Annual Cryptosporidium-attributable MSD incidence was 3.48 (95% CI, 2.27–4.67) and 3.18 (95% CI, 1.85–4.52) per 100 child-years in African and Asian infants, respectively, and 1.41 (95% CI, 0.73–2.08) and 1.36 (95% CI, 0.66–2.05) per 100 child-years in toddlers. Corresponding Cryptosporidium-attributable LSD incidences per 100 child-years were 2.52 (95% CI, 0.33–5.01) and 4.88 (95% CI, 0.82–8.92) in infants and 4.04 (95% CI, 0.56–7.51) and 4.71 (95% CI, 0.24–9.18) in toddlers. We estimate 2.9 and 4.7 million Cryptosporidium-attributable cases annually in children aged <24 months in the sub-Saharan Africa and India/Pakistan/Bangladesh/Nepal/Afghanistan regions, respectively, and ~202,000 Cryptosporidium-attributable deaths (regions combined). ~59,000 excess deaths occurred among Cryptosporidium-attributable diarrhea cases over expected if cases had been Cryptosporidium-negative. Conclusions The enormous African/Asian Cryptosporidium disease burden warrants investments to develop vaccines, diagnostics and therapies. PMID:27219054

Intestinal protozoa in HIV-infected patients in Apulia, South Italy.

PubMed Central

Brandonisio, O.; Maggi, P.; Panaro, M. A.; Lisi, S.; Andriola, A.; Acquafredda, A.; Angarano, G.

1999-01-01

Protozoa are important enteric pathogens in patients with human immunodeficiency virus (HIV) infection. In this study the prevalence of intestinal protozoa in 154 HIV-infected patients, with or without diarrhoea, in our region (Apulia, South Italy) was evaluated between December 1993 and February 1998. In the majority of patients CD4+ T cell count was below 200/microl. The overall prevalence of intestinal protozoa was 43/154 (27.92%). Twenty-eight (43.08%) out of 65 patients with diarrhoea and 15 (16-85%) out of 89 non-diarrhoeic patients were parasitized. In particular, in the group of 65 patients with diarrhoea the following protozoa were identified: Cryptosporidium parvum in 14 (21.54%), Blastocystis hominis in 7 (10.77%), microsporidia in 6 (9.23%), Giardia lamblia in 4 (6.15%) and Isospora belli in 1 (1.54%). Three patients were Cryptosporidium parvum-microsporidia co-infected. In patients without intestinal symptoms, prevalence was 3/89 (3.37%) for Cryptosporidium parvum, 9/89 (10.11%) for Blastocystis hominis, 1/89 (1.12%) for microsporidia and 2/89 (2.25%) for Giardia lamblia. A significant (P<0.001) correlation was observed between protozoan infection and the presence of diarrhoea. In particular, Cryptosporidium parvum and microsporidia infections were significantly (P<0.001) and P = 0.046, respectively) associated with diarrhoeal illness. Moreover, the majority of cases of cryptosporidiosis were first diagnosed in the periods of heaviest rainfall. Therefore, drinking water contamination may be a possible source of human infection in our area. PMID:10694157

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

Diversity of microsporidia, Cryptosporidium and Giardia in mountain gorillas (Gorilla beringei beringei) in Volcanoes National Park, Rwanda.

PubMed

Sak, Bohumil; Petrželková, Klára J; Květoňová, Dana; Mynářová, Anna; Pomajbíková, Kateřina; Modrý, David; Cranfield, Michael R; Mudakikwa, Antoine; Kváč, Martin

2014-01-01

Infectious diseases represent the greatest threats to endangered species, and transmission from humans to wildlife under increased anthropogenic pressure has been always stated as a major risk of habituation. To evaluate the impact of close contact with humans on the occurrence of potentially zoonotic protists in great apes, one hundred mountain gorillas (Gorilla beringei beringei) from seven groups habituated either for tourism or for research in Volcanoes National Park, Rwanda were screened for the presence of microsporidia, Cryptosporidium spp. and Giardia spp. using molecular diagnostics. The most frequently detected parasites were Enterocytozoon bieneusi found in 18 samples (including genotype EbpA, D, C, gorilla 2 and five novel genotypes gorilla 4-8) and Encephalitozoon cuniculi with genotype II being more prevalent (10 cases) compared to genotype I (1 case). Cryptosporidium muris (2 cases) and C. meleagridis (2 cases) were documented in great apes for the first time. Cryptosporidium sp. infections were identified only in research groups and occurrence of E. cuniculi in research groups was significantly higher in comparison to tourist groups. No difference in prevalence of E. bieneusi was observed between research and tourist groups. Although our data showed the presence and diversity of important opportunistic protists in Volcanoes gorillas, the source and the routes of the circulation remain unknown. Repeated individual sampling, broad sampling of other hosts sharing the habitat with gorillas and quantification of studied protists would be necessary to acquire more complex data.

Prevalence and Multilocus Genotyping Analysis of Cryptosporidium and Giardia Isolates from Dogs in Chiang Mai, Thailand.

PubMed

Tangtrongsup, Sahatchai; Scorza, A Valeria; Reif, John S; Ballweber, Lora R; Lappin, Michael R; Salman, Mo D

2017-05-10

The occurrence and zoonotic potential of Cryptosporidium spp. and Giardia duodenalis isolated from dogs in Chiang Mai, Thailand were determined. Fecal samples were collected from 109 dogs between July and August 2008. Cryptosporidium spp. infection was determined by immunofluorescent assay (IFA), PCR assays that amplify Cryptosporidium heat-shock protein 70 kDa (hsp70), and two PCR assays that amplify a small subunit-ribosomal RNA (SSU-rRNA). Giardia duodenalis infection was identified using zinc sulfate centrifugal flotation, IFA, and four PCR assays that amplify the Giardia glutamate dehydrogenase (gdh), beta-giardin (bg), and generic and dog-specific assays of triosephosphate isomerase (tpi) genes. Overall prevalence of Cryptosporidium spp. and G. duodenalis was 31.2% and 45.9%, respectively. Sequence analysis of 22 Cryptosporidium -positive samples and 21 Giardia -positive samples revealed the presence of C. canis in 15, and C. parvum in 7, G. duodenalis Assemblage C in 8, D in 11, and mixed of C and D in 2 dogs. Dogs in Chiang Mai were commonly exposed to Cryptosporidium spp. and G. duodenalis . Cryptosporidium parvum can be isolated from the feces of dogs, and all G. duodenalis assemblages were dog-specific. Dogs could be a reservoir for a zoonotic Cryptosporidium infection in humans, but further studies will be required to determine the clinical and zoonotic importance.

Prevalence and Multilocus Genotyping Analysis of Cryptosporidium and Giardia Isolates from Dogs in Chiang Mai, Thailand

PubMed Central

Tangtrongsup, Sahatchai; Scorza, A. Valeria; Reif, John S.; Ballweber, Lora R.; Lappin, Michael R.; Salman, Mo D.

2017-01-01

The occurrence and zoonotic potential of Cryptosporidium spp. and Giardia duodenalis isolated from dogs in Chiang Mai, Thailand were determined. Fecal samples were collected from 109 dogs between July and August 2008. Cryptosporidium spp. infection was determined by immunofluorescent assay (IFA), PCR assays that amplify Cryptosporidium heat-shock protein 70 kDa (hsp70), and two PCR assays that amplify a small subunit-ribosomal RNA (SSU-rRNA). Giardia duodenalis infection was identified using zinc sulfate centrifugal flotation, IFA, and four PCR assays that amplify the Giardia glutamate dehydrogenase (gdh), beta-giardin (bg), and generic and dog-specific assays of triosephosphate isomerase (tpi) genes. Overall prevalence of Cryptosporidium spp. and G. duodenalis was 31.2% and 45.9%, respectively. Sequence analysis of 22 Cryptosporidium-positive samples and 21 Giardia-positive samples revealed the presence of C. canis in 15, and C. parvum in 7, G. duodenalis Assemblage C in 8, D in 11, and mixed of C and D in 2 dogs. Dogs in Chiang Mai were commonly exposed to Cryptosporidium spp. and G. duodenalis. Cryptosporidium parvum can be isolated from the feces of dogs, and all G. duodenalis assemblages were dog-specific. Dogs could be a reservoir for a zoonotic Cryptosporidium infection in humans, but further studies will be required to determine the clinical and zoonotic importance. PMID:29056685

Detection and molecular characterization of Cryptosporidium and Eimeria species in Philippine bats.

PubMed

Murakoshi, Fumi; Recuenco, Frances C; Omatsu, Tsutomu; Sano, Kaori; Taniguchi, Satoshi; Masangkay, Joseph S; Alviola, Philip; Eres, Eduardo; Cosico, Edison; Alvarez, James; Une, Yumi; Kyuwa, Shigeru; Sugiura, Yuki; Kato, Kentaro

2016-05-01

The genus Cryptosporidium, which is an obligate intracellular parasite, infects various vertebrates and causes a diarrheal disease known as cryptosporidiosis. Bats are naturally infected with zoonotic pathogens; thus, they are potential reservoirs of parasites. We investigated the species and genotype distribution as well as prevalence of Cryptosporidium and Eimeria in Philippine bats. We captured and examined 45 bats; four were positive for Cryptosporidium spp. and seven were positive for Eimeria spp. We detected Cryptosporidium bat genotype II from Ptenochirus jagori. Three other Cryptosporidium sequences, detected from Rhinolophus inops, Cynopterus brachyotis, and Eonycteris spelaea, could not be classified as any known species or genotype; we therefore propose the novel genotype Cryptosporidium bat genotypes V, VI, and VII. Bat genotype V is associated with human cryptosporidiosis clade, and therefore, this genotype may be transmissible to humans. Among the Eimeria sequences, BE3 detected from Scotophilus kuhlii was classified with known bat and rodent clades; however, other sequences detected from C. brachyotis, E. spelaea, Rousettus amplexicaudatus, and R. inops could not be classified with known Eimeria species. These isolates might represent a new genotype. Our findings demonstrate that the bats of the Philippines represent a reservoir of multiple Cryptosporidium and Eimeria spp.

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.

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

Cryptosporidium meleagridis in an Indian ring-necked parrot (Psittacula krameri).

PubMed

Morgan, U M; Xiao, L; Limor, J; Gelis, S; Raidal, S R; Fayer, R; Lal, A; Elliot, A; Thompson, R C

2000-03-01

To perform a morphological and genetic characterisation of a Cryptosporidium infection in an Indian ring-necked parrot (Psittacula krameri) and to compare this with C meleagridis from a turkey. Tissue and intestinal sections from an Indian ring-necked parrot were examined microscopically for Cryptosporidium. The organism was also purified from the crop and intestine, the DNA extracted and a portion of the 18S rDNA gene amplified, sequenced and compared with sequence and biological information obtained for C meleagridis from a turkey as well as sequence information for other species of Cryptosporidium. Morphological examination of tissue sections from an Indian ring-necked parrot revealed large numbers of Cryptosporidium oocysts attached to the apical border of enterocytes lining the intestinal tract. Purified Cryptosporidium oocysts measured about 5.1 x 4.5 microns, which conformed morphologically to C meleagridis. The sequence obtained from this isolate was identical to sequence information obtained from a C meleagridis isolate from a turkey. Cryptosporidium meleagridis was detected in an Indian ring-necked parrot using morphological and molecular methods. This is the first time that this species of Cryptosporidium has been reported in a non-galliform host and extends the known host range of C meleagridis.

Microbial pathogens in source and treated waters from drinking water treatment plants in the United States and implications for human health

USGS Publications Warehouse

King, Dawn N.; Donohue, Maura J.; Vesper, Stephen J.; Villegas, Eric N.; Ware, Michael W.; Vogel, Megan E.; Furlong, Edward; Kolpin, Dana W.; Glassmeyer, Susan T.; Pfaller, Stacy

2016-01-01

An occurrence survey was conducted on selected pathogens in source and treated drinking water collected from 25 drinking water treatment plants (DWTPs) in the United States. Water samples were analyzed for the protozoa Giardia and Cryptosporidium (EPA Method 1623); the fungi Aspergillus fumigatus, Aspergillus niger and Aspergillus terreus (quantitative PCR [qPCR]); and the bacteria Legionella pneumophila (qPCR), Mycobacterium avium, M. avium subspecies paratuberculosis, and Mycobacterium intracellulare (qPCR and culture). Cryptosporidium and Giardia were detected in 25% and in 46% of the source water samples, respectively (treated waters were not tested). Aspergillus fumigatus was the most commonly detected fungus in source waters (48%) but none of the three fungi were detected in treated water. Legionella pneumophila was detected in 25% of the source water samples but in only 4% of treated water samples. M. avium and M. intracellulare were both detected in 25% of source water, while all three mycobacteria were detected in 36% of treated water samples. Five species of mycobacteria, Mycobacterium mucogenicum, Mycobacterium phocaicum, Mycobacterium triplex, Mycobacterium fortuitum, and Mycobacterium lentiflavum were cultured from treated water samples. Although these DWTPs represent a fraction of those in the U.S., the results suggest that many of these pathogens are widespread in source waters but that treatment is generally effective in reducing them to below detection limits. The one exception is the mycobacteria, which were commonly detected in treated water, even when not detected in source waters.

Prevalence and Genotyping of Cryptosporidium Infection in Pet Parrots in North China

PubMed Central

Zhang, Xiao-Xuan; Zhang, Nian-Zhang; Zhao, Guang-Hui; Zhao, Quan; Zhu, Xing-Quan

2015-01-01

Cryptosporidiosis is a worldwide zoonosis caused by Cryptosporidium spp., sometimes leading to severe diarrhea in humans and animals. In the present study, 311 parrots, belonging to four species, namely, Budgerigars (Melopsittacus undulatus), Lovebirds (Agapornis sp.), Alexandrine parakeets (Psittacula eupatria), and Cockatiel (Nymphicus hollandicus), from Beijing and Weifang cities, were examined for Cryptosporidium spp. infection. Blood samples of each bird were examined using enzyme linked immunosorbent assay (ELISA) and fecal samples were examined by Sheather's sugar flotation technique. Prevalence of Cryptosporidium infection were 3.22% (10/311) and 0.64% (2/311) by ELISA and Sheather's sugar flotation technique, respectively. Seroprevalence of Cryptosporidium infection in different breeds varied from 0 to 15.39%. Sequencing analysis showed that both positive samples from fecal samples belonged to Cryptosporidium avian genotype V. This is the first report of Cryptosporidium avian genotype V in Budgerigars. The results of the present study provided foundation-data for prevention and control of cryptosporidiosis in pet birds in China. PMID:26273629

Prevalence and Genotyping of Cryptosporidium Infection in Pet Parrots in North China.

PubMed

Zhang, Xiao-Xuan; Zhang, Nian-Zhang; Zhao, Guang-Hui; Zhao, Quan; Zhu, Xing-Quan

2015-01-01

Cryptosporidiosis is a worldwide zoonosis caused by Cryptosporidium spp., sometimes leading to severe diarrhea in humans and animals. In the present study, 311 parrots, belonging to four species, namely, Budgerigars (Melopsittacus undulatus), Lovebirds (Agapornis sp.), Alexandrine parakeets (Psittacula eupatria), and Cockatiel (Nymphicus hollandicus), from Beijing and Weifang cities, were examined for Cryptosporidium spp. infection. Blood samples of each bird were examined using enzyme linked immunosorbent assay (ELISA) and fecal samples were examined by Sheather's sugar flotation technique. Prevalence of Cryptosporidium infection were 3.22% (10/311) and 0.64% (2/311) by ELISA and Sheather's sugar flotation technique, respectively. Seroprevalence of Cryptosporidium infection in different breeds varied from 0 to 15.39%. Sequencing analysis showed that both positive samples from fecal samples belonged to Cryptosporidium avian genotype V. This is the first report of Cryptosporidium avian genotype V in Budgerigars. The results of the present study provided foundation-data for prevention and control of cryptosporidiosis in pet birds in China.

Molecular characterisation of a disseminated Cryptosporidium infection in a Koi carp (Cyprinus carpio).

PubMed

Yang, Rongchang; Dorrestein, Gerry M; Ryan, Una

2016-08-15

Cryptosporidium is a protozoan parasite that infects a wide range of hosts, yet relatively little is known about the epidemiology of cryptosporidiosis in fish. Here we report a disseminated Cryptosporidium infection in a male Koi carp (Cyprinus carpio), with parasite stages identified deep within the epithelium of the intestine, kidneys, spleen, liver and gills causing severe granulomatous inflammatory lesions. Molecular characterization at two loci; 18S ribosomal RNA (rRNA) and actin, revealed this to be a novel Cryptosporidium genotype, most closely related to Cryptosporidium molnari. Copyright © 2016 Elsevier B.V. All rights reserved.

Criteria For Evaluation of Proposed Protozoan Detection Methods

EPA Science Inventory

Currently, the only EPA approved method for detection and quantitation of protozoan cysts and oöcysts in source and drinking water, is the “ICR Protozoan Method for Detecting Giardia Cysts and Cryptosporidium Oöcysts in Water by a Fluorescent Antibody Procedure (ICR Microbial La...

Irrigation waters as a source of pathogenic microorganisms in produce: a review

USDA-ARS?s Scientific Manuscript database

There is increasing evidence that consumption of raw fresh produce is a major factor contributing to human gastrointestinal illness. A wide variety of pathogens contribute to food-borne illnesses, including bacteria (e.g., Salmonella, pathogenic E. coli), protozoa (e.g., Cryptosporidium, Giardia), ...

The potential for zoonotic transmission of Giardia duodenalis and Cryptosporidium spp. from beef and dairy cattle in Ontario, Canada.

PubMed

Dixon, Brent; Parrington, Lorna; Cook, Angela; Pintar, Katarina; Pollari, Frank; Kelton, David; Farber, Jeffrey

2011-01-10

The objective of this study was to compare the occurrence and the genotypes and species of Giardia duodenalis and Cryptosporidium spp. in beef and dairy cattle from farms in the Regional Municipality of Waterloo, Ontario, in an effort to determine the potential for zoonotic transmission from these animals. Pooled manure samples were collected from 45 dairy cattle farms and 30 beef cattle farms. The presence of Giardia cysts and Cryptosporidium oocysts was determined by immunofluorescence microscopy, while nested-PCR and DNA sequencing were used to determine genotypes and species. The overall farm prevalence was very high for both Giardia and Cryptosporidium, and was similar for dairy cattle farms (96 and 64%, respectively) and beef cattle farms (97 and 63%, respectively). However, on dairy cattle farms, G. duodenalis and Cryptosporidium spp. were detected in 44% and 6% of total pooled pen manure samples, respectively, with the occurrence of both parasites being generally higher in calves than in older animals. Most Giardia isolates were identified as either the host-adapted genotype G. duodenalis Assemblage E or the zoonotic Assemblage B. Cryptosporidium parvum and Cryptosporidium andersoni were the most frequently identified species in dairy cattle, while the non-zoonotic species Cryptosporidium ryanae and Cryptosporidium bovis were also found. On beef cattle farms, 72% and 27% of the total pooled pen manure samples were positive for Giardia and Cryptosporidium, respectively, with no obvious correlation with age. All Giardia isolates in beef cattle were identified as G. duodenalis Assemblage E, while all Cryptosporidium isolates were identified by sequence analysis as C. andersoni, although microscopic analyses, and subsequent restriction fragment length polymorphism analyses, indicated that other Cryptosporidium species were also present. The results of this study indicate that although Giardia and Cryptosporidium were identified in a higher overall percentage of the pooled beef cattle manure samples than in dairy cattle, firmly established zoonotic genotypes and species were much more common in dairy cattle than in beef cattle in this region. Dairy cattle, and especially dairy calves, may, therefore, pose a greater risk of infection to humans than beef cattle. However, these results may also provide evidence of potential zooanthroponotic transmission (human to animal). Crown Copyright © 2010. Published by Elsevier B.V. All rights reserved.

Occurrence and molecular characterization of Cryptosporidium spp. and Enterocytozoon bieneusi in dairy cattle, beef cattle and water buffaloes in China.

PubMed

Ma, Jingbo; Li, Pei; Zhao, Xiaoping; Xu, Hailing; Wu, Wenxian; Wang, Yuanfei; Guo, Yaqiong; Wang, Lin; Feng, Yaoyu; Xiao, Lihua

2015-01-30

Cryptosporidium spp. and Enterocytozoon bieneusi are important protists in a wide range of vertebrate hosts, causing diarrheal diseases. Cattle are considered potential reservoirs of Cryptosporidium infection in humans, although their role in the transmission of E. bieneusi is not clear. In the present work, 793 fecal specimens from dairy cattle, native beef cattle, and water buffaloes on 11 farms in China were examined for the presence of Cryptosporidium spp. and E. bieneusi using nested PCR targeting the small subunit (SSU) rRNA gene of Cryptosporidium spp. and the internal transcribed spacer (ITS) of E. bieneusi. For Cryptosporidium, 144/446 (32.3%) dairy cattle, 44/166 (26.5%) beef cattle, and 43/181 (23.8%) water buffaloes were PCR-positive. Sequence analysis was successful for 213 of the 231 Cryptosporidium-positive isolates; among them 94 had Cryptosporidium andersoni, 61 had Cryptosporidium bovis, 54 had Cryptosporidium ryanae, 2 had a Cryptosporidium suis-like genotype, and 2 had mixed infections of C. bovis and C. ryanae. In dairy and beef cattle, C. andersoni and C. bovis were the most common species, whereas C. ryanae was the dominant species in water buffaloes. The latter species produced SSU rRNA sequences different between cattle and water buffaloes. For E. bieneusi, the infection rate of E. bieneusi in dairy cattle, beef cattle and water buffaloes was 4.9%, 5.4% and 2.2%, respectively. All 35 E. bieneusi-positive specimens were successfully sequenced, revealing the presence of four genotypes: three Group 2 genotypes previously reported in cattle as well as humans (I, J and BEB4) and one Group 1 genotype recently reported in yaks (CHN11). Genotypes I and J were the most common genotypes in dairy and beef cattle, while genotype CHN11 was the only genotype seen in water buffaloes. Thus, the distribution of Cryptosporidium spp. and E. bieneusi in water buffaloes might be different from in dairy and beef cattle in China. These findings indicate that some of the Cryptosporidium species and all four E. bieneusi genotypes identified in bovine animals in the study areas may have zoonotic potential. Copyright © 2014 Elsevier B.V. All rights reserved.

Molecular detection and characterization of Cryptosporidium species in household dogs, pet shop puppies, and dogs kept in a school of veterinary nursing in Japan.

PubMed

Itoh, Naoyuki; Oohashi, Yoshino; Ichikawa-Seki, Madoka; Itagaki, Tadashi; Ito, Yoichi; Saeki, Hideharu; Kanai, Kazutaka; Chikazawa, Seishiro; Hori, Yasutomo; Hoshi, Fumio; Higuchi, Seiichi

2014-03-01

Members of Cryptosporidium species, which are protozoan parasites, are prevalent worldwide and can cause diarrhoea in both humans and animals, including dogs. In addition, the Cryptosporidium species harboured in dogs have the potential for zoonotic transmission. The purpose of the present study was to determine the prevalence of Cryptosporidium species infection and perform molecular characterization of isolates in household dogs, pet shop puppies, and dogs kept in a school of veterinary nursing in Japan. Fresh faecal samples were collected once from 529 household dogs (aged from 2 months to 18 years old, from 9 veterinary clinics located in 6 different regions), 471 pet shop puppies (≤ 3 months old, from 4 pet shops located in 2 different regions), and 98 dogs (aged from 2 to 11 years old) kept in a veterinary nursing school. A nested polymerase chain reaction (PCR) assay targeting the 18S rRNA gene was employed for the detection of Cryptosporidium species, and 111 random samples of PCR amplicons (approximately 500-bp) were sequenced for the molecular characterization of the isolates. The prevalences of Cryptosporidium species in household dogs, pet shop puppies, and veterinary nursing school dogs were 7.2%, 31.6%, and 18.4%, respectively. In household dogs, no significant correlation was observed between the prevalence of Cryptosporidium species and the age (≤ 6 months vs. >6 months), living conditions (indoor vs. outdoor), faecal conditions (formed vs. unformed), and location of residence. In pet shop puppies, the prevalence of Cryptosporidium species was not related to faecal condition; however, the prevalence significantly differed among the pet shops. All of the 111 sequence samples (26 from household dogs, 75 from pet shop puppies, and 10 from veterinary nursing school dogs) were identified as Cryptosporidium canis. The present study demonstrates a high prevalence of Cryptosporidium species infections in pet shop puppies and dogs of a veterinary nursing school in Japan. However, because Cryptosporidium hominis and Cryptosporidium parvum are the most common causes of human infections, it is likely that the risk of zoonotic transmission of Cryptosporidium species from dogs to humans is low. Copyright © 2014 Elsevier B.V. All rights reserved.

Contribution of treated wastewater to the contamination of recreational river areas with Cryptosporidium spp. and Giardia duodenalis.

PubMed

Castro-Hermida, José Antonio; García-Presedo, Ignacio; Almeida, André; González-Warleta, Marta; Correia Da Costa, José Manuel; Mezo, Mercedes

2008-07-01

Samples of the influent and final effluent from 12 wastewater treatment plants from Galicia (NW, Spain) were analyzed for the presence of Cryptosporidium spp. oocysts and Giardia duodenalis cysts. All of the plants discharge effluent to a hydrographic basin in which there are numerous recreational areas and fluvial beaches. The samples (25-50 liters) were collected in spring, summer, autumn and winter of 2007. A total of 96 samples were analyzed using techniques included in the US Environmental Protection Agency Method 1623. To identify the genotypes present, the following genes were amplified and sequenced: 18S SSU rRNA (Cryptosporidium spp.) and beta-giardina (G. duodenalis). Both parasites were detected in influent and effluent samples from all treatment plants (100%) throughout the year, and G. duodenalis always outnumbered Cryptosporidium spp. The mean concentration of G. duodenalis per liter of influent was significantly higher (P<0.05) than the mean concentration of Cryptosporidium spp. per liter of influent. The mean concentrations of parasites in influent samples ranged from 6 to 350 Cryptosporidium spp. oocysts per liter and from 89 to 8305 G. duodenalis cysts per liter. In final treated effluent, the mean concentration of parasites ranged from 2 to 390 Cryptosporidium spp. oocysts per liter and from 79 to 2469 G. duodenalis cysts per liter. The distribution of results per season revealed that in all plants, the highest number of (oo)cysts were detected in spring and summer. Cryptosporidium parvum, Cryptosporidium andersoni, Cryptosporidium hominis and assemblages A-I, A-II, E of G. duodenalis were detected. The risk of contamination of water courses by Cryptosporidium spp. and G. duodenalis is therefore considerable. It is important that wastewater treatment authorities reconsider the relevance of the levels of contamination by both parasites in wastewater, and develop adequate countermeasures.

Hollow-fiber ultrafiltration of Cryptosporidium parvum oocysts from a wide variety of 10-L surface water samples.

PubMed

Kuhn, Ryan C; Oshima, Kevin H

2002-06-01

An optimized hollow-fiber ultrafiltration system (50 000 MWCO) was developed to concentrate Cryptosporidium oocysts from 10-L samples of environmental water. Seeded experiments were conducted using a number of surface-water samples from the southwestern U.S.A. and source water from four water districts with histories of poor oocyst recovery. Ultrafiltration produced a mean recovery of 47.9% from 19 water samples (55.3% from 39 individual tests). We also compared oocyst recoveries using the hollow-fiber ultrafiltration system with those using the Envirochek filter. In limited comparison tests, the hollow-fiber ultrafiltration system produced recoveries similar to those of the Envirochek filter (hollow fiber, 74.1% (SD = 2.8); Envirochek, 71.9% (SD = 5.2)) in low-turbidity (3.9 NTU) samples and performed better than the Envirochek filter in high-turbidity (159.0 NTU) samples (hollow fiber, 27.5%; Envirochek, 0.4%). These results indicate that hollow-fiber ultrafiltration can efficiently recover oocysts from a wide variety of surface waters and may be a cost-effective alternative for concentrating Cryptosporidium from water, given the reusable nature of the filter.

Survey on gastrointestinal parasites and detection of Cryptosporidium spp. on cattle in West Java, Indonesia.

PubMed

Ananta, Sylvia Maharani; Suharno; Hidayat, Adi; Matsubayashi, Makoto

2014-03-01

To evaluate the presence of gastrointestinal parasites on cattle in Indonesia because the prevalence of parasites varies between countries depending on the terrain surrounding livestock farms and investigations in Indonesia have never been performed. Fecal samples from cattle at 35 farms in 7 districts in West Java, Indonesia, has been examined using the floatation or sedimentation methods, and a immunofluorescence assay and experimentally inoculation to mice for Cryptosporidium or Giardia.spp. 153 of 394 examined cattle (38.8%) were infected with gastrointestinal parasites. The prevalence of Eimeria spp., Nematoda spp. (including Oesophagustomum and Bunostomum-like), Fasciola gigantica and Paramphistomum spp. was 22.4%, 11.2%, 12.5% and 3.8%, respectively. Cryptosporidium andersoni (C. andersoni) was also found in two samples. One isolate of this parasite was confirmed to be transmitted to mice, in contrast to the isolates from other countries. although this survey is preliminary, the results shows that the infection of gastrointestinal parasites in Indonesia was not high, but these infected cattle could be as a potential source leading to economic losses in livestock production. Copyright © 2014 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

Genetic diversity of Cryptosporidium in fish at the 18S and actin loci and high levels of mixed infections.

PubMed

Yang, Rongchang; Palermo, Cindy; Chen, Linda; Edwards, Amanda; Paparini, Andrea; Tong, Kaising; Gibson-Kueh, Susan; Lymbery, Alan; Ryan, Una

2015-12-15

Cryptosporidium is an enteric parasite that infects humans and a wide range of animals. Relatively little is known about the epidemiology and taxonomy of Cryptosporidium in fish. In the present study, a total of 775 fish, belonging to 46 species and comprising ornamental fish, marine fish and freshwater fish were screened for the prevalence of Cryptosporidium by PCR. The overall prevalence of Cryptosporidium in fish was 5.3% (41/775), with prevalences ranging from 1.5 to 100% within individual host species. Phylogenetic analysis of these Cryptosporidium isolates as well as 14 isolates from previous studies indicated extensive genetic diversity as well as evidence for mixed infections. At the 18S locus the following species were identified; Cryptosporidium molnari-like genotype (n=14), Cryptosporidium huwi (n=8), piscine genotype 2 (n=4), piscine genotype 3-like (n=1), piscine genotype 4 (n=2), piscine genotype 5 (n=13), piscine genotype 5-like (n=1) and five novel genotypes (n=5). At the actin locus, species identification agreed with the 18S locus for only 52.3% of isolates sequenced, indicating high levels of mixed infections. Future studies will need to employ both morphological characterization and deep sequencing amplicon-based technologies to better understand the epidemiological and phylogenetic relationships of piscine-derived Cryptosporidium species and genotypes, particularly when mixed infections are detected. Copyright © 2015 Elsevier B.V. All rights reserved.

Comparison of Assays for Sensitive and Reproducible Detection of Cell Culture-Infectious Cryptosporidium parvum and Cryptosporidium hominis in Drinking Water

PubMed Central

Di Giovanni, George D.; Rochelle, Paul A.

2012-01-01

This study compared the three most commonly used assays for detecting Cryptosporidium sp. infections in cell culture: immunofluorescent antibody and microscopy assay (IFA), PCR targeting Cryptosporidium sp.-specific DNA, and reverse transcriptase PCR (RT-PCR) targeting Cryptosporidium sp.-specific mRNA. Monolayers of HCT-8 cells, grown in 8-well chamber slides or 96-well plates, were inoculated with a variety of viable and inactivated oocysts to assess assay performance. All assays detected infection with low doses of flow cytometry-enumerated Cryptosporidium parvum oocysts, including infection with one oocyst and three oocysts. All methods also detected infection with Cryptosporidium hominis. The RT-PCR assay, IFA, and PCR assay detected infection in 23%, 25%, and 51% of monolayers inoculated with three C. parvum oocysts and 10%, 9%, and 16% of monolayers inoculated with one oocyst, respectively. The PCR assay was the most sensitive, but it had the highest frequency of false positives with mock-infected cells and inactivated oocysts. IFA was the only infection detection assay that did not produce false positives with mock-infected monolayers. IFA was also the only assay that detected infections in all experiments with spiked oocysts recovered from Envirochek capsules following filtration of 1,000 liters of treated water. Consequently, cell culture with IFA detection is the most appropriate method for routine and sensitive detection of infectious Cryptosporidium parvum and Cryptosporidium hominis in drinking water. PMID:22038611

Zoonotic Cryptosporidium Species and Enterocytozoon bieneusi Genotypes in HIV-Positive Patients on Antiretroviral Therapy

PubMed Central

Wang, Lin; Zhang, Hongwei; Zhao, Xudong; Zhang, Longxian; Zhang, Guoqing; Guo, Meijin; Liu, Lili; Xiao, Lihua

2013-01-01

Molecular diagnostic tools have been used increasingly in the characterization of the transmission of cryptosporidiosis and microsporidiosis in developing countries. However, few studies have examined the distribution of Cryptosporidium species and Enterocytozoon bieneusi genotypes in AIDS patients receiving antiretroviral therapy. In the present study, 683 HIV-positive patients in the National Free Antiretroviral Therapy Program in China and 683 matched HIV-negative controls were enrolled. Cryptosporidium species and subtypes and Enterocytozoon bieneusi genotypes were detected and differentiated by PCR and DNA sequencing. The infection rates were 1.5% and 0.15% for Cryptosporidium and 5.7% and 4.2% for E. bieneusi in HIV-positive and HIV-negative participants, respectively. The majority (8/11) of Cryptosporidium cases were infections by zoonotic species, including Cryptosporidium meleagridis (5), Cryptosporidium parvum (2), and Cryptosporidium suis (1). Prevalent E. bieneusi genotypes detected, including EbpC (39), D (12), and type IV (7), were also potentially zoonotic. The common occurrence of EbpC was a feature of E. bieneusi transmission not seen in other areas. Contact with animals was a risk factor for both cryptosporidiosis and microsporidiosis. The results suggest that zoonotic transmission was significant in the epidemiology of both diseases in rural AIDS patients in China. PMID:23224097

Microscopic and Molecular Detection of Cryptosporidium andersoni and Cryptosporidium xiaoi in Wastewater Samples of Tehran Province, Iran

PubMed Central

HATAM-NAHAVANDI, Kareem; MOHEBALI, Mehdi; MAHVI, Amir-Hossein; KESHAVARZ, Hossein; NAJAFIAN, Hamid-Reza; MIRJALALI, Hamed; REZAEI, Sasan; REZAEIAN, Mostafa

2016-01-01

Background: As a waterborne pathogen, Cryptosporidium is one of the most common causes of gastroenteritis in human and hoofed livestock animals. This study aimed to investigate the distribution of Cryptosporidium spp. in human and livestock wastewaters in Iran, by the 18S rRNA sequence analysis. Methods: A total of 54 raw wastewater samples collected from three urban treatment plants and two slaughterhouses during 2014–2015 in Tehran, Iran. The presence of the Cryptosporidium oocysts was assessed by immunofluorescence with monoclonal antibodies. To characterize the oocysts at the molecular level, the 18S rRNA gene of Cryptosporidium was PCR amplified and sequenced. Results: Of the 54 wastewater samples examined, 34 (62.9%) were positive for Cryptosporidium oocysts using the IFA. Of these, 70.5% (24/34) were positive by PCR, that 91.6% (22/24) were successfully sequenced. The species of C. andersoni (95.4%) and C. xiaoi (4.6%) were detected in livestock wastewater samples. Conclusion: C. andersoni was the major Cryptosporidium sp. found in the aquatic environmental wastewater samples. The high rate of detection of C. andersoni in domestic wastewater was probably the result of the predominancy of this species in cattle herds in Iran. The current study is the first report of C. xiaoi in Iran. PMID:28127361

Prevalence, transmission, and host specificity of Cryptosporidium spp. in various animal groups from two French zoos.

PubMed

Osman, Marwan; El Safadi, Dima; Benamrouz-Vanneste, Sadia; Cian, Amandine; Moriniere, Romain; Gantois, Nausicaa; Delgado-Viscogliosi, Pilar; Guyot, Karine; Bosc, Stéphanie; Chabé, Magali; Petit, Thierry; Viscogliosi, Eric; Certad, Gabriela

2017-12-01

Cryptosporidium represents a major cause of gastrointestinal illness in humans and animals including domestic, wild, and in captivity animals, and more than 30 validated species of Cryptosporidium are recognized as infectious to different hosts such as mammals, birds, reptiles, amphibians, and fish. Therefore, numerous investigations have been conducted worldwide in order to shed light on the epidemiology of this parasite and to explore its potential reservoirs. Few surveys, targeting humans and animals have been carried out regarding the epidemiology of Cryptosporidium spp. in France and no data are available about the circulation of this parasite in French zoological gardens. Herein, we determined the prevalence of Cryptosporidium in animals housed in two French zoos. A total of 307 fecal samples belonging to 161 species were screened by nested PCR. Overall, Cryptosporidium DNA was detected in 1.9% of the 161 species and 1% of the total number of fecal samples tested. Additionally, three Cryptosporidium species were identified: C. galli, C. andersoni, and C. tyzzeri. To our knowledge, this is the first molecular study focused on Cryptosporidium infection in captivity animals in France. This study is of interest considering the exposure of a large number of humans and animals to this waterborne protozoan, found ubiquitously in the environment.

Molecular characterization of a Cryptosporidium isolate from a banded mongoose Mungos mungo.

PubMed

Abe, Niichiro; Takami, Kazutoshi; Kimata, Isao; Iseki, Motohiro

2004-02-01

Cryptosporidium spp. has been found in more than 150 species of mammals, but there has been no report in mongooses. In this study, we report the isolation of Cryptosporidium sp. in a banded mongoose Mungos mungo, which was brought from Tanzania to Japan; the isolate was analyzed genetically to validate the occurrence of a new, host-adapted genotype. Cryptosporidium diagnostic fragments of 18S ribosomal RNA and 70-kDa heat shock protein genes were amplified from this isolate and compared with the other Cryptosporidium species and genotypes reported previously. Analyses showed that the mongoose isolate represents a new genotype, closely related to that of bears.

DETECTION OF PROTOZOAN PARASITES IN SOURCE AND FINISHED WATER - 3RD EDITION ASM'S METHODS IN ENVIRONMENTAL MICROBIOLOGY

EPA Science Inventory

Protozoans are eukaryotic organisms which can live either a free-living or parasitic existence. Some free-living forms, under the right conditions, can become opportunistic parasites. Enteric pathogenic protozoans, like Giardia and Cryptosporidium, which are now known to be tra...

40 CFR 141.521 - What updated watershed control requirements must my unfiltered system implement to continue to...

Code of Federal Regulations, 2010 CFR

2010-07-01

... Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Filtration and Disinfection-Systems Serving Fewer Than 10,000... oocysts in the source water. Your system's watershed control program must, for Cryptosporidium: (a...

40 CFR 141.521 - What updated watershed control requirements must my unfiltered system implement to continue to...

Code of Federal Regulations, 2012 CFR

2012-07-01

... Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Filtration and Disinfection-Systems Serving Fewer Than 10,000... oocysts in the source water. Your system's watershed control program must, for Cryptosporidium: (a...

40 CFR 141.521 - What updated watershed control requirements must my unfiltered system implement to continue to...

Code of Federal Regulations, 2014 CFR

2014-07-01

... Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Filtration and Disinfection-Systems Serving Fewer Than 10,000... oocysts in the source water. Your system's watershed control program must, for Cryptosporidium: (a...

40 CFR 141.521 - What updated watershed control requirements must my unfiltered system implement to continue to...

Code of Federal Regulations, 2013 CFR

2013-07-01

... Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Filtration and Disinfection-Systems Serving Fewer Than 10,000... oocysts in the source water. Your system's watershed control program must, for Cryptosporidium: (a...

No molecular epidemiological evidence supporting household transmission of zoonotic Giardia duodenalis and Cryptosporidium spp. from pet dogs and cats in the province of Álava, Northern Spain.

PubMed

de Lucio, Aida; Bailo, Begoña; Aguilera, María; Cardona, Guillermo A; Fernández-Crespo, Juan C; Carmena, David

2017-06-01

The role of pet dogs and cats as suitable source of human infections by the diarrheagenic protozoan parasites Giardia duodenalis and Cryptosporidium spp. has been a topic of intense debate for long time and still remains a largely unsolved problem. In this cross-sectional molecular epidemiological survey we attempted to investigate whether zoonotic (or zooanthroponotic) disease transmission was occurring among humans and domestic dogs and cats sharing the same spatial and temporal setting in both rural and urban areas of the province of Álava, Northern Spain. A total of 268 (including 179 human, 55 canine, and 34 feline) individual faecal specimens were obtained from 63 family households during February-March and November-December 2014. Detection of G. duodenalis cysts and Cryptosporidium spp. oocysts was achieved by direct fluorescence microscopy (DFAT) and PCR-based methods targeting the small subunit (SSU) ribosomal RNA gene of the parasites. Giardia-positive isolates were subsequently sub-genotyped at the glutamate dehydrogenase (GDH) and β-giardin (BG) genes. Overall, G. duodenalis infections were identified in 3.4% (6/179) of humans, 29% (16/55) of dogs, and 5.9% (2/34) of cats, respectively. Cryptosporidium spp. infections were detected in 1.1% (2/179) of humans, 5.5% (3/55) of dogs, and 8.8% (3/34) of cats, respectively. Simultaneous infections in human and canine/feline hosts by G. duodenalis or Cryptosporidium spp. were only demonstrated in a single household in which a cat and its owner tested positive for Cryptosporidium by DFAT, but this result could not be confirmed by SSU-PCR. Infections were homogeneously distributed among the studied human or animal populations irrespectively of their sex, age group, or geographical region of origin. Inadequate washing of raw vegetables and fruits was the only risk factor significantly associated to a higher likelihood of having human giardiosis/cryptosporidiosis. Molecular characterization of G. duodenalis isolates revealed the presence of sub-assemblage BIV in a single human isolate. All dog (n=3) and cat (n=2) isolates successfully genotyped were assigned to canine- and feline-specific assemblages C and F, respectively. No mixed assemblage or sub-assemblage infections could be demonstrated. Regarding Cryptosporidium, C. canis was found infecting dogs (n=2), and C. felis a single cat. Attempts to amplify and characterize Cryptosporidium human isolates failed repeatedly. Our results suggest that pet dogs and cats do not seem to play a significant role as suitable reservoirs of human giardiosis or cryptosporidiosis in the province of Álava. We conclude, therefore, that zoonotic transmission of giardiosis or cryptosporidiosis among pet dogs and cats and their owners in this geographical region is very likely a rare event. Copyright © 2017 Elsevier B.V. All rights reserved.

Cryptosporidium Contamination and Attributed Risks in Yunlong Lake in Xuzhou, China

PubMed Central

Kong, Yadong; Yuan, Tao; Niu, Jinghui; Li, Zhaoji; Yang, Baisong

2017-01-01

Swimming in surface water bodies (e.g., lakes, rivers) can expose the human body to substantial risk of infection by Cryptosporidium. These findings are from a one-year investigation on the occurrence and distribution of the protozoan parasite Cryptosporidium in Yunlong Lake, Xuzhou, China. Cryptosporidium oocysts were detected by immunofluorescence microscopy. From January to November of 2015, 180 samples (120 water samples and 60 sediment samples) were collected and analyzed. Among them, 42 (35%) water samples and 28 (47%) sediment samples tested positive for Cryptosporidium. The concentration of Cryptosporidium oocysts in the water samples was 0–8/10 L and 0–260/g in sediment samples. Results revealed that July was the highest risk period for both swimming and diving with an estimated probability of infection from swimming of greater than 18 per 10,000 swim sessions. It was concluded that swimming or diving in Yunlong Lake has a higher risk of Cryptosporidium infection than the acceptable risk level set by the United States Environmental Protection Agency. Thus, regular monitoring of water quality in recreation water bodies is strongly recommended. PMID:28386287

Prevalence of Cryptosporidium sp. infection in diarrheic and non-diarrheic humans in Iran

PubMed Central

2007-01-01

For evaluation of the prevalence of Cryptosporidium sp. infection in diarrheic and non-diarrheic humans in Iran, fecal specimens from diarrheic (n = 129) and non-diarrheic humans (n = 271) were collected and examined for the presence of Cryptosporidium sp. oocysts. The presence of Cryptosporidium sp. oocysts was determined by Ziehl-Neelsen acid-fast staining. Humans were grouped according to their age as follows: younger than 15, 16-25, 26-35, 36-50, and over 51 years. The results showed that the overall prevalence of infection in all 400 samples was 10.8%, but the prevalence (25.6%) in diarrheic humans was higher than that (3.7%) in non-diarrheic humans. Oocysts of Cryptosporidium sp. were detected in the feces of 21.4%, 9.3%, 8.8%, 6.7% and 5.7% of different age groups, respectively. The intensity of oocysts was significantly higher in diarrheic humans than in non-diarrheic ones. There was a significant association between Cryptosporidium sp. infection and occurrence of diarrhea (P < 0.05). The results indicate that Cryptosporidium sp. infection is prevalent in diarrheic humans in Iran. PMID:17570977

Issues of the presence of parasitic protozoa in surface waters

NASA Astrophysics Data System (ADS)

Hawrylik, Eliza

2018-02-01

Parasitic protozoa are very numerous organisms in the environment that play an important role in the spread of water-borne diseases. Water-borne epidemics caused by parasitic protozoa are noted throughout the world. Within these organisms, intestinal protozoa of the genera Cryptosporidium and Giardia are ones of the most serious health hazards for humans. This paper focuses on the problem of the presence of parasitic protozoa in surface waters. Characteristics of the most frequently recognized pathogens responsible for water-borne outbreaks were described, as well as sources of contamination and surface waters contamination due to protozoa of the genus Cryptosporidium and Giardia were presented. The methods of destroying the cysts and oocysts of parasitic protozoa used nowadays in the world were also presented in a review.

Contamination of River Water by Cryptosporidium parvum Oocysts in Western Japan

PubMed Central

Ono, Kazuo; Tsuji, Hidetaka; Rai, Shiba Kumar; Yamamoto, Akio; Masuda, Kuniyoshi; Endo, Takuro; Hotta, Hak; Kawamura, Takashi; Uga, Shoji

2001-01-01

In Japan, only a few rivers have been inspected for Cryptosporidium parvum contamination, and the methods used had low sensitivity. In 1998 and 1999, we used a method with higher sensitivity to examine all large rivers used as sources of water supply in one prefecture (which we divided into four areas) in western Japan for Cryptosporidium oocysts. One sample was collected at each of 156 sites along 18 rivers, and samples were tested for Cryptosporidium oocysts by immunomagnetic separation. Samples were classified as being obtained on an island with livestock and fishing industries, a densely populated urban area, a western region including farming villages, or a still more rural northern area with agriculture and fishing. Restriction fragment length polymorphism analysis was used for identification of the C. parvum found as the bovine or human type. C. parvum was detected in at least one sample from 13 of the 18 rivers and in 47% (74 of 156) of the samples. One-third to all of the samples from each area contained C. parvum oocysts. The number of C. parvum oocysts per 20 liters of river water varied in the same pattern as the number of cattle kept in the four kinds of areas (as determined by the Mantel extension test). Oocysts isolated were of the bovine type; the C. parvum detected in rivers probably came from cattle kept in that valley. As we had expected, when tested with a more sensitive method, river water in western Japan was found to be greatly contaminated with C. parvum oocysts, as reported in other countries. PMID:11525974

Role of Wall Shear Stress in Cryptosporidium parvum Oocyst Attachment to Environmental Biofilms.

PubMed

Luo, Xia; Jedlicka, Sabrina S; Jellison, Kristen L

2017-12-15

This study investigated Cryptosporidium parvum oocyst deposition onto biofilms as a function of shear stress under laminar or turbulent flow. Annular rotating bioreactors were used to grow stabilized stream biofilms at shear stresses ranging from 0.038 to 0.46 Pa. These steady-state biofilms were then used to assess the impact of hydrodynamic conditions on C. parvum oocyst attachment. C. parvum deposition onto biofilms followed a pseudo-second-order model under both laminar (after a lag phase) and turbulent flows. The total number of oocysts attached to the biofilm at steady state decreased as the hydrodynamic wall shear stress increased. The oocyst deposition rate constant increased with shear stress but decreased at high shear, suggesting that increasing wall shear stress results in faster attachment of Cryptosporidium due to higher mass transport until the shear forces exceed a critical limit that prevents oocyst attachment. These data show that oocyst attachment in the short and long term are impacted differently by shear: higher shear (to a certain limit) may be associated with faster initial oocyst attachment, but lower shear is associated with greater numbers of oocysts attached at equilibrium. IMPORTANCE This research provides experimental evidence to demonstrate that shear stress plays a critical role in protozoan-pathogen transport and deposition in environmental waters. The data presented in this work expand scientific understanding of Cryptosporidium attachment and fate, which will further influence the development of timely and accurate sampling strategies, as well as advanced water treatment technologies, to target protozoan pathogens in surface waters that serve as municipal drinking water sources. Copyright © 2017 American Society for Microbiology.

RESEARCH IN FILTRATION FOR CRYPTOSPORIDIUM REMOVAL

EPA Science Inventory

The USEPA has conducted pilot plant studies for the removal of Cryptosporidium oocysts from drinking water. Fourteen pilot-scale tests were performed to assess the ability of conventional treatment to control Cryptosporidium oocysts and three surrogates; turbidity, total particle...

Cryptosporidium species from common edible bivalves in Manila Bay, Philippines.

PubMed

Pagoso, Edison Jay A; Rivera, Windell L

2017-06-15

Manila Bay is one of the major propagation sites of edible bivalves in the Philippines. Studies have shown that bivalves might be contaminated with human pathogens like the protozoan parasite Cryptosporidium, one of the major causes of gastroenteritis in the world. In this study, Cryptosporidium from four species of edible bivalves were isolated using a combination of sucrose flotation and immunomagnetic separation. Using direct fluorescent antibody test, Cryptosporidium oocysts were found in 67 out of 144 samples collected. DNA sequence analysis of the 18S rRNA gene of the isolates detected C. parvum and C. hominis (major causes of human cryptosporidiosis) and C. meleagridis (causes infection in avian species). Analysis of the 60kDa glycoprotein gene further confirmed the genotypes of the Cryptosporidium isolates. This study is the first to provide baseline information on Cryptosporidium contamination of Manila Bay where bivalves are commonly cultured. Copyright © 2017 Elsevier Ltd. All rights reserved.

New Insights into Human Cryptosporidiosis

PubMed Central

Clark, Douglas P.

1999-01-01

Cryptosporidium parvum is an important cause of diarrhea worldwide. Cryptosporidium causes a potentially life-threatening disease in people with AIDS and contributes significantly to morbidity among children in developing countries. In immunocompetent adults, Cryptosporidium is often associated with waterborne outbreaks of acute diarrheal illness. Recent studies with human volunteers have indicated that Cryptosporidium is highly infectious. Diagnosis of infection with this parasite has relied on identification of acid-fast oocysts in stool; however, new immunoassays or PCR-based assays may increase the sensitivity of detection. Although the mechanism by which Cryptosporidium causes diarrhea is still poorly understood, the parasite and the immune response to it probably combine to impair absorption and enhance secretion within the intestinal tract. Important genetic studies suggest that humans can be infected by at least two genetically distinct types of Cryptosporidium, which may vary in virulence. This may, in part, explain the clinical variability seen in patients with cryptosporidiosis. PMID:10515902

Prevalence of pathogenic free-living amoeba and other protozoa in natural and communal piped tap water from Queen Elizabeth protected area, Uganda.

PubMed

Sente, Celsus; Erume, Joseph; Naigaga, Irene; Mulindwa, Julius; Ochwo, Sylvester; Magambo, Phillip Kimuda; Namara, Benigna Gabriela; Kato, Charles Drago; Sebyatika, George; Muwonge, Kevin; Ocaido, Michael

2016-08-03

Pathogenic water dwelling protozoa such as Acanthamoeba spp., Hartmannella spp., Naegleria spp., Cryptosporidium spp. and Giardia spp. are often responsible for devastating illnesses especially in children and immunocompromised individuals, yet their presence and prevalence in certain environment in sub-Saharan Africa is still unknown to most researchers, public health officials and medical practitioners. The objective of this study was to establish the presence and prevalence of pathogenic free-living amoeba (FLA), Cryptosporidium and Giardia in Queen Elizabeth Protected Area (QEPA). Samples were collected from communal taps and natural water sites in QEPA. Physical water parameters were measured in situ. The samples were processed to detect the presence of FLA trophozoites by xenic cultivation, Cryptosporidium oocysts by Ziehl-Neelsen stain and Giardia cysts by Zinc Sulphate floatation technique. Parasites were observed microscopically, identified, counted and recorded. For FLA, genomic DNA was extracted for amplification and sequencing. Both natural and tap water sources were contaminated with FLA, Cryptosporidium spp. and Giardia spp. All protozoan parasites were more abundant in the colder rainy season except for Harmannella spp. and Naegleria spp. which occurred more in the warmer months. The prevalence of all parasites was higher in tap water than in natural water samples. There was a strong negative correlation between the presence of Acanthamoeba spp., Hartmannella spp., Cryptosporidium spp. and Giardia spp. with Dissolved Oxygen (DO) (P < 0.05). The presence of Cryptosporidium spp. showed a significant positive correlation (P < 0.05) with conductivity, pH and Total Dissolved Solids (TDS); whereas the presence of Giardia spp. had only a strong positive correlation with TDS. Molecular genotyping of FLA produced 7 Acanthamoeba, 5 Echinamoeba, 2 Hartmannella, 1 Bodomorpha, 1 Nuclearia and 1 Cercomonas partial sequences. All water collection sites were found to be contaminated with pathogenic protozoa that could possibly be the cause of a number of silent morbidities and mortalities among rural households in QEPA. This implies that water used by communities in QEPA is of poor quality and predisposes them to a variety of protozoan infections including the FLA whose public health importance was never reported, thus necessitating adoption of proper water safety measures.

Sensitivity, specificity and comparison of three commercially available immunological tests in the diagnosis of Cryptosporidium species in animals.

PubMed

Danišová, Olga; Halánová, Monika; Valenčáková, Alexandra; Luptáková, Lenka

The study was conducted to compare the specificity of immunological diagnostic methods used for the diagnosis of Cryptosporidium species capable of causing life-threatening infection in both immunosuppressed and immunocompetent patients. For the detection of Cryptosporidium species in 79 animals with diarrhoea, we used three Copro-antigen tests: RIDASCREEN ® Cryptosporidium test, Cryptosporidium 2nd Generation (ELISA) and RIDA ® QUICK Cryptosporidium. For immunoassays we used positive and negative samples detected by means of polymerase chain reaction and validated by sequencing and nested polymerase chain reaction to confirm the presence six different species of Cryptosporidium species. Prevalence of cryptosporidiosis in the entire group determined by enzyme immunoassay, enzyme linked immunosorbent assay, immuno-chromatographic test and polymerase chain reaction was 34.17%, 27.84%, 6.33% and 27.84%, respectively. Sensitivity of animal samples with enzyme immunoassay, enzyme linked immunosorbent assay, and immuno-chromatographic test was 63.6%, 40.9% and 22.7%, resp., when questionable samples were considered positive, whereas specificity of enzyme immunoassay, enzyme linked immunosorbent assay and immuno-chromatographic test was 75.9%, 78.9% and 100%, respectively. Positive predictive values and negative predictive values were different for all the tests. These differences results are controversial and therefore reliability and reproducibility of immunoassays as the only diagnostic method is questionable. The use of various Cryptosporidium species in diagnosis based on immunological testing and different results obtained by individual tests indicate potential differences in Copro-antigens produced by individual Cryptosporidium species. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

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. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Evolution of monitoring for Giardia and Cryptosporidium in water.

PubMed

Efstratiou, Artemis; Ongerth, Jerry; Karanis, Panagiotis

2017-10-15

This review describes the evolution of monitoring methodology for Cryptosporidium and Giardia in water since the 1970's. Methods in current use for Giardia and Cryptosporidium in water are highlighted, though attention is given to all available published methods by country and continent. The review is intended to stimulate research leading to future improvements and further developments in monitoring methodology for Giardia, Cryptosporidium and other waterborne protozoan parasites in water. Copyright © 2017. Published by Elsevier Ltd.

Modulation of the immune response by infection with Cryptosporidium spp. in children with allergic diseases.

PubMed

Guangorena-Gómez, J O; Maravilla-Domínguez, A; García-Arenas, G; Cervantes-Flores, M; Meza-Velázquez, R; Rivera-Guillén, M; Acosta-Saavedra, L C; Goytia-Acevedo, R C

2016-08-01

It has been demonstrated that the allergic response can be ameliorated by the administration of pathogen derivatives that activate Toll-like receptors and induce a Th1-type immune response (IR). Cryptosporidium is a parasite that promotes an IR via Toll-like receptors and elicits the production of Th1-type cytokines, which limit cryptosporidiosis. The aim of this study was to investigate allergy-related immune markers in children naturally infected with Cryptosporidium. In a cross-sectional study, 49 children with or without clinical diagnosis of allergies, oocysts of Cryptosporidium spp. in the faeces were screened microscopically. We microscopically screened for leucocytes, examined T and B cells for allergy-related activation markers using flow cytometry and evaluated serum for total IgE using chemiluminescence. Children with allergies and Cryptosporidium in the faeces had significantly lower levels of total IgE, B cells, CD19(+) CD23(+) and CD19(+) CD124(+) cells as well as a greater percentage of interferon-gamma (IFN-γ(+) ) and IL-4(+) CD4(+) cells than children with allergies without Cryptosporidium. This is the first description of the modulation of the IR in children with allergic diseases in the setting of natural Cryptosporidium infection. Our findings suggest the involvement of CD4(+) cells producing IL-4 and IFN-γ in the IR to Cryptosporidium in naturally infected children. © 2016 John Wiley & Sons Ltd.

Molecular identification of the Cryptosporidium deer genotype in the Hokkaido sika deer (Cervus nippon yesoensis) in Hokkaido, Japan.

PubMed

Kato, Satomi; Yanagawa, Yojiro; Matsuyama, Ryota; Suzuki, Masatsugu; Sugimoto, Chihiro

2016-04-01

The protozoan Cryptosporidium occurs in a wide range of animal species including many Cervidae species. Fecal samples collected from the Hokkaido sika deer (Cervus nippon yesoensis), a native deer of Hokkaido, in the central, western, and eastern areas of Hokkaido were examined by polymerase chain reaction (PCR) to detect infections with Cryptosporidium and for sequence analyses to reveal the molecular characteristics of the amplified DNA. DNA was extracted from 319 fecal samples and examined with PCR using primers for small-subunit ribosomal RNA (SSU-rRNA), actin, and 70-kDa heat shock protein (HSP70) gene loci. PCR-amplified fragments were sequenced and phylogenetic trees were created. In 319 fecal samples, 25 samples (7.8 %) were positive with SSU-rRNA PCR that were identified as the Cryptosporidium deer genotype. Among Cryptosporidium-positive samples, fawns showed higher prevalence (16.1 %) than yearlings (6.4 %) and adults (4.7 %). The result of Fisher's exact test showed a statistical significance in the prevalence of the Cryptosporidium deer genotype between fawn and other age groups. Sequence analyses with actin and HSP70 gene fragments confirmed the SSU-rRNA result, and there were no sequence diversities observed. The Cryptosporidium deer genotype appears to be the prevalent Cryptosporidium species in the wild sika deer in Hokkaido, Japan.

Microbial pathogens in source and treated waters from drinking water treatment plants in the United States and implications for human health.

PubMed

King, Dawn N; Donohue, Maura J; Vesper, Stephen J; Villegas, Eric N; Ware, Michael W; Vogel, Megan E; Furlong, Edward F; Kolpin, Dana W; Glassmeyer, Susan T; Pfaller, Stacy

2016-08-15

An occurrence survey was conducted on selected pathogens in source and treated drinking water collected from 25 drinking water treatment plants (DWTPs) in the United States. Water samples were analyzed for the protozoa Giardia and Cryptosporidium (EPA Method 1623); the fungi Aspergillus fumigatus, Aspergillus niger and Aspergillus terreus (quantitative PCR [qPCR]); and the bacteria Legionella pneumophila (qPCR), Mycobacterium avium, M. avium subspecies paratuberculosis, and Mycobacterium intracellulare (qPCR and culture). Cryptosporidium and Giardia were detected in 25% and in 46% of the source water samples, respectively (treated waters were not tested). Aspergillus fumigatus was the most commonly detected fungus in source waters (48%) but none of the three fungi were detected in treated water. Legionella pneumophila was detected in 25% of the source water samples but in only 4% of treated water samples. M. avium and M. intracellulare were both detected in 25% of source water, while all three mycobacteria were detected in 36% of treated water samples. Five species of mycobacteria, Mycobacterium mucogenicum, Mycobacterium phocaicum, Mycobacterium triplex, Mycobacterium fortuitum, and Mycobacterium lentiflavum were cultured from treated water samples. Although these DWTPs represent a fraction of those in the U.S., the results suggest that many of these pathogens are widespread in source waters but that treatment is generally effective in reducing them to below detection limits. The one exception is the mycobacteria, which were commonly detected in treated water, even when not detected in source waters. Published by Elsevier B.V.

Cryptosporidium ubiquitum n.sp. in animals and humans

USDA-ARS?s Scientific Manuscript database

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...

Molecular epidemiology of Cryptosporidium in HIV/AIDS patients in Malaysia.

PubMed

Asma, I; Sim, B L H; Brent, R D; Johari, S; Yvonne Lim, A L

2015-06-01

Cryptosporidiosis is a particular concern in immunocompromised individuals where symptoms may be severe. The aim of this study was to examine the epidemiological and molecular characteristics of Cryptosporidium infections in HIV/AIDS patients in Malaysia in order to identify risk factors and facilitate control measures. A modified Ziehl-Neelsen acid fast staining method was used to test for the presence of Cryptosporidium oocysts in the stools of 346 HIV/AIDS patients in Malaysia. Standard coproscopical methods were used to identify infections with other protozoan or helminths parasites. To identify the species of Cryptosporidium, DNA was extracted and nested-PCR was used to amplify a portion of the SSU rRNA gene. A total of 43 (12.4%) HIV-infected patients were found to be infected with Cryptosporidium spp. Of the 43 Cryptosporidium-positive HIV patients, 10 (23.3%) also harboured other protozoa, and 15 (34.9%) had both protozoa and helminths. The highest rates of cryptosporidiosis were found in adult males of Malay background, intravenous drug users, and those with low CD4 T cell counts (i.e., < 200 cells/mm3). Most were asymptomatic and had concurrent opportunistic infections mainly with Mycobacterium tuberculosis. DNA sequence analysis of 32 Cryptosporidium isolates identified C. parvum (84.3%), C. hominis (6.3%), C. meleagridis (6.3%), and C. felis (3.1%). The results of the present study revealed a high prevalence of Cryptosporidium infection in hospitalized HIV/AIDS patients. The results also confirmed the potential significance of zoonotic transmission of C. parvum in HIV infected patients, as it was the predominant species found in this study. However, these patients were found to be susceptible to a wide range of Cryptosporidium species. Epidemiological and molecular characterization of Cryptosporidium isolates provides clinicians and researchers with further information regarding the origin of the infection, and may enhance treatment and control strategies.

Genotyping and subtyping of Giardia and Cryptosporidium isolates from commensal rodents in China.

PubMed

Zhao, Z; Wang, R; Zhao, W; Qi, M; Zhao, J; Zhang, L; Li, J; Liu, A

2015-05-01

Cryptosporidium and Giardia are two important zoonotic intestinal parasites responsible for diarrhoea in humans and other animals worldwide. Rodents, as reservoirs or carriers of Cryptosporidium and Giardia, are abundant and globally widespread. In the present study, we collected 232 fecal specimens from commensal rodents captured in animal farms and farm neighbourhoods in China. We collected 33 Asian house rats, 168 brown rats and 31 house mice. 6.0% (14/232) and 8.2% (19/232) of these rodents were microscopy-positive for Giardia cysts and Cryptosporidium oocysts, respectively. All 14 Giardia isolates were identified as Giardia duodenalis assemblage G at a minimum of one or maximum of three gene loci (tpi, gdh and bg). By small subunit rRNA (SSU rRNA) gene sequencing, Cryptosporidium parvum (n = 12) and Cryptosporidium muris (n = 7) were identified. The gp60 gene encoding the 60-kDa glycoprotein was successfully amplified and sequenced in nine C. parvum isolates, all of which belonged to the IIdA15G1 subtype. Observation of the same IIdA15G1 subtype in humans (previously) and in rodents (here) suggests that rodents infected with Cryptosporidium have the potential to transmit cryptosporidiosis to humans.

Cryptosporidium erinacei n. sp. (Apicomplexa: Cryptosporidiidae) in hedgehogs.

PubMed

Kváč, Martin; Hofmannová, Lada; Hlásková, Lenka; Květoňová, Dana; Vítovec, Jiří; McEvoy, John; Sak, Bohumil

2014-03-17

The morphological, biological, and molecular characteristics of Cryptosporidium hedgehog genotype are described, and the species name Cryptosporidium erinacei n. sp. is proposed to reflect its specificity for hedgehogs under natural and experimental conditions. Oocysts of C. erinacei are morphologically indistinguishable from Cryptosporidium parvum, measuring 4.5-5.8 μm (mean=4.9 μm) × 4.0-4.8 μm (mean=4.4 μm) with a length to width ratio of 1.13 (1.02-1.35) (n=100). Oocysts of C. erinacei obtained from a naturally infected European hedgehog (Erinaceus europaeus) were infectious for naïve 8-week-old four-toed hedgehogs (Atelerix albiventris); the prepatent period was 4-5 days post infection (DPI) and the patent period was longer than 20 days. C. erinacei was not infectious for 8-week-old SCID and BALB/c mice (Mus musculus), Mongolian gerbils (Meriones unguiculatus), or golden hamsters (Mesocricetus auratus). Phylogenetic analyses based on small subunit rRNA, 60 kDa glycoprotein, actin, Cryptosporidium oocyst wall protein, thrombospondin-related adhesive protein of Cryptosporidium-1, and heat shock protein 70 gene sequences revealed that C. erinacei is genetically distinct from previously described Cryptosporidium species. Copyright © 2014 Elsevier B.V. All rights reserved.

Giardia and Cryptosporidium species and genotypes in coyotes (Canis latrans).

PubMed

Trout, James M; Santín, Mónica; Fayer, Ronald

2006-06-01

Feces and duodenal scrapings were collected from 22 coyotes (Canis latrans) killed in managed hunts in northeastern Pennsylvania. Polymerase chain reaction (PCR) methods were used to detect Giardia and Cryptosporidium spp. PCR-amplified fragments of Giardia and Cryptosporidium spp. SSU-rRNA genes were subjected to DNA sequence analysis for species/genotype determination. Seven coyotes (32%) were positive for G. duodenalis: three assemblage C, three assemblage D, and one assemblage B. Six coyotes (27%) were positive for Cryptosporidium spp. One isolate shared 99.7% homology with C. muris, whereas five others (23%) shared 100% homology with C. canis, coyote genotype. This is the first report on multiple genotypes of Giardia spp. in coyotes and on the prevalence of Cryptosporidium spp. genotypes in coyotes.

Occurrence of Cryptosporidium spp. in Antillean manatees (Trichechus manatus) and Amazonian manatees (Trichechus inunguis) from Brazil.

PubMed

Borges, Joāo Carlos Gomes; Alves, Leucio Câmara; Faustino, Maria Aparecida da Gloria; Marmontel, Miriam

2011-12-01

Infections by Cryptosporidium spp. in aquatic mammals is a major concern due to the possibility of the waterborne transmission of oocysts. The aim of the present study was to report the occurrence of Cryptosporidium spp. in Antillean manatees (Trichechus manatus) and Amazonian manatees (Trichechus inunguis) from Brazil. Fecal samples were collected and processed using Kinyoun's method. Positive samples were also submitted to the direct immunofluorescence test. The results revealed the presence of Cryptosporidium spp. oocysts in 12.5% (17/136) of the material obtained from the Antillean manatees and in 4.3% (05/115) of the samples from the Amazonian manatees. Cryptosporidium spp. infection was more prevalent in captive animals than in free-ranging specimens.

40 CFR 141.704 - Analytical methods.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 22 2010-07-01 2010-07-01 false Analytical methods. 141.704 Section... Monitoring Requirements § 141.704 Analytical methods. (a) Cryptosporidium. Systems must analyze for Cryptosporidium using Method 1623: Cryptosporidium and Giardia in Water by Filtration/IMS/FA, 2005, United States...

40 CFR 141.704 - Analytical methods.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 23 2014-07-01 2014-07-01 false Analytical methods. 141.704 Section... Monitoring Requirements § 141.704 Analytical methods. (a) Cryptosporidium. Systems must analyze for Cryptosporidium using Method 1623: Cryptosporidium and Giardia in Water by Filtration/IMS/FA, 2005, United States...

40 CFR 141.704 - Analytical methods.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 24 2013-07-01 2013-07-01 false Analytical methods. 141.704 Section... Monitoring Requirements § 141.704 Analytical methods. (a) Cryptosporidium. Systems must analyze for Cryptosporidium using Method 1623: Cryptosporidium and Giardia in Water by Filtration/IMS/FA, 2005, United States...

40 CFR 141.704 - Analytical methods.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 23 2011-07-01 2011-07-01 false Analytical methods. 141.704 Section... Monitoring Requirements § 141.704 Analytical methods. (a) Cryptosporidium. Systems must analyze for Cryptosporidium using Method 1623: Cryptosporidium and Giardia in Water by Filtration/IMS/FA, 2005, United States...

40 CFR 141.704 - Analytical methods.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 24 2012-07-01 2012-07-01 false Analytical methods. 141.704 Section... Monitoring Requirements § 141.704 Analytical methods. (a) Cryptosporidium. Systems must analyze for Cryptosporidium using Method 1623: Cryptosporidium and Giardia in Water by Filtration/IMS/FA, 2005, United States...

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...

GENOTYPES OF CRYPTOSPORIDIUM INFECTING FUR-BEARING MAMMALS DIFFER FROM THOSE INFECTING HUMANS

EPA Science Inventory

To evaluate the public health significance of wildlife Cryptosporidium spp., five species of small wild mammals (beavers, muskrats, otters, raccoons, and foxes) living in Chesapeake Bay watersheds were examined in this study for the presence of Cryptosporidium spp., using a small...

Molecular analysis of Cryptosporidium from cattle from five states of Peninsular Malaysia.

PubMed

Yap, Nan Jiun; Koehler, Anson V; Ebner, Janine; Tan, Tiong Kai; Lim, Yvonne A L; Gasser, Robin B

2016-02-01

Despite the importance of the cattle industry in Malaysia, there are very few studies of the diversity and public health significance of bovine cryptosporidiosis in this country. In the present study, we used a PCR-based approach to detect and genetically characterize Cryptosporidium DNA in faecal samples from a cohort of 215 asymptomatic cattle (of different ages) from six farms from five states of Peninsular Malaysia. Cattle on four of the six farms were test-positive for Cryptosporidium, with an overall prevalence of 3.2%. Cryptosporidium bovis and Cryptosporidium ryanae were detected in two (0.9%) and five (2.3%) samples tested; this low prevalence likely relates to the age of the cattle tested, as most (73%) of the samples tested originated from cattle that were ≥2 years of age. Future studies should investigate the zoonotic potential of Cryptosporidium in pre-weaned and weaned calves in rural communities of Malaysia. Copyright © 2016 Elsevier Ltd. All rights reserved.

Giardia and Cryptosporidium in cetaceans on the European Atlantic coast.

PubMed

Reboredo-Fernández, Aurora; Ares-Mazás, Elvira; Martínez-Cedeira, José A; Romero-Suances, Rafael; Cacciò, Simone M; Gómez-Couso, Hipólito

2015-02-01

The occurrence of Giardia and Cryptosporidium was investigated in cetacean specimens stranded on the northwestern coast of Spain (European Atlantic coast) by analysis of 65 samples of large intestine from eight species. The parasites were identified by direct immunofluorescence antibody test (IFAT) and by PCR amplification of the β-giardin gene, the ITS1-5.8S-ITS2 region and the SSU-rDNA gene of Giardia and the SSU-rDNA gene of Cryptosporidium. Giardia and Cryptosporidium were detected in 7 (10.8 %) and 9 samples (13.8 %), respectively. In two samples, co-infection with both parasites was observed. Giardia duodenalis assemblages A, C, D and F, and Cryptosporidium parvum were identified. This is the first report of G. duodenalis in Balaenoptera acutorostrata, Kogia breviceps and Stenella coeruleoalba and also the first report of Cryptosporidium sp. in B. acutorostrata and of C. parvum in S. coeruleoalba and Tursiops truncatus. These results extend the known host range of these waterborne enteroparasites.

Multiplex PCR detection of waterborne intestinal protozoa: microsporidia, Cyclospora, and Cryptosporidium.

PubMed

Lee, Seung-Hyun; Joung, Migyo; Yoon, Sejoung; Choi, Kyoungjin; Park, Woo-Yoon; Yu, Jae-Ran

2010-12-01

Recently, emerging waterborne protozoa, such as microsporidia, Cyclospora, and Cryptosporidium, have become a challenge to human health worldwide. Rapid, simple, and economical detection methods for these major waterborne protozoa in environmental and clinical samples are necessary to control infection and improve public health. In the present study, we developed a multiplex PCR test that is able to detect all these 3 major waterborne protozoa at the same time. Detection limits of the multiplex PCR method ranged from 10(1) to 10(2) oocysts or spores. The primers for microsporidia or Cryptosporidium used in this study can detect both Enterocytozoon bieneusi and Encephalitozoon intestinalis, or both Cryptosporidium hominis and Cryptosporidium parvum, respectively. Restriction enzyme digestion of PCR products with BsaBI or BsiEI makes it possible to distinguish the 2 species of microsporidia or Cryptosporidium, respectively. This simple, rapid, and cost-effective multiplex PCR method will be useful for detecting outbreaks or sporadic cases of waterborne protozoa infections.

[Molecular Detection of Giardia lamblia and Cryptosporidium Species in Pet Dogs].

PubMed

GU, You-fang; WANG, Kai; LIU, De-yi; MEI, Nan; CHEN, Cheng; CHEN, Tao; HAN, Min-min; ZHOU, Li; CAO, Jia-tong; ZHANG, Heng; ZHANG, Xue-liang; FAN, Zi-lai; LI, Wen-chao

2015-10-01

To determine the prevalence of Giardia lamblia and Cryptosporidium species infection in pet dogs, and identify the G. lamblia assemblages and Cryptosporidium species. A total of 315 fresh fecal samples were collected from pet clinics in five counties of Anhui Province and in Hangzhou of Zhejiang Province. Hemi-nested-PCR targeting the GDH gene of G. lamblia and nested-PCR targeting the SSU rRNA gene of Cryptosporidium were performed in all the fecal samples. The PCR products were sequenced and analyzed using bioinformatics methods to identify the G. lamblia assemblages and Cryptosporidium species. The positive rates of G. lamblia and Cryptosporidium spp. infections in the 315 fecal samples were 3.2% (10/315) and 1.6% (5/315), respectively. Specifically, the two indicators were both significantly higher in dogs ≤12 months (17.8% and 11.1%, respectively) than in adult dogs (0.7% and 0.0%)(P<0.05). However, there was no significant difference in the two indicators between male and female dogs. In addition, two G. lamblia assemblages were identified, assemblages B (n=6) and D (n=4). Sequence analysis of PCR products of the SSU rRNA gene showed that the five Cryptosporidium isolates were C. canis (n =5). The prevalences of G. lamblia and Cryptosporidium infection in pet dogs in Anhui and Zhejiang Provinces were 3.2 % and 1.6 %, respectively. The assemblages of G. lamblia in this study are of types B and D.

The molecular characterisation of Cryptosporidium species in relinquished dogs in Great Britain: a novel zoonotic risk?

PubMed

Rosanowski, S M; Banica, M; Ellis, E; Farrow, E; Harwood, C; Jordan, B; James, C; McKenna, D; Fox, M; Blake, D P

2018-05-01

Surveillance was conducted to investigate the occurrence of protozoan parasites of the genus Cryptosporidium in dogs newly admitted to a dog rehoming charity in London, Great Britain. Voided faecal samples were collected from all new admissions between 2011 and 2012 during six separate 4-week sampling periods. Information on host signalment, including age, breed and reason for submission and faecal consistency, was collected. Polymerase Chain Reaction (PCR) targeting the 18S ribosomal RNA gene, confirmed by sequencing, was conducted on the faecal samples to detect Cryptosporidium genomic DNA and determine Cryptosporidium identity. In total, 677 dogs were included in the study. The prevalence of Cryptosporidium-positive faecal samples was 4.6% (31/676). There were positive samples in all of the six sampling periods. Cryptosporidium canis (n = 28), C. parvum (n = 2) and C. andersoni (n = 1) were identified. Sixty KDa glycoprotein (gp60) gene amplicon sequencing of the C. parvum samples identified genotypes IIaA17G1R1 and IIaA15G2R1 for the first time from a dog. There were no significant associations between signalment data and Cryptosporidium status. While this was a study of one rehoming shelter, the presence of the potentially zoonotic C. parvum and C. canis in dogs highlights a public health concern. Further research is needed to better understand the epidemiology and potential impacts of Cryptosporidium infection in dogs.

Molecular detection of Giardia duodenalis and Cryptosporidium spp. in canine faecal samples contaminating public areas in Northern Italy.

PubMed

Simonato, Giulia; Frangipane di Regalbono, Antonio; Cassini, Rudi; Traversa, Donato; Tessarin, Cinzia; Di Cesare, Angela; Pietrobelli, Mario

2017-12-01

Giardia duodenalis and Cryptosporidium spp. are common intestinal pathogens of humans and animals. Dogs may be infected by zoonotic isolates of G. duodenalis and Cryptosporidium spp. that, consequently, have high interest under public health perspective. This study estimated the occurrence of these protozoa in canine faeces polluting public areas of Padua municipality (Northern Italy), towards a potential evaluation of health risks for dogs and humans. A total of 705 canine stools was collected in green (n = 270) and urban (n = 435) areas and processed by duplex real-time PCR and real-time PCR SYBR® Green I for the detection of both protozoa. Positive samples were submitted to specific nested PCRs (i.e. β-giardin/SSU-rRNA genes for Giardia; SSU-rRNA gene for Cryptosporidium) to obtain detailed information on the isolates retrieved.Giardia and Cryptosporidium prevalence were 28.9% and 1.7%, respectively. Twenty-one Giardia-positive samples were successfully identified as dog-specific assemblages C and D, and 1 as the human-specific assemblage B. One isolate was identified as Cryptosporidium canis, while the other 11 were confirmed to belong to the Cryptosporidium parvum species complex. Contrariwise to the Cryptosporidium low prevalence, the wide distribution of Giardia suggests a high risk of infection for dogs attending public areas. Although data indicate a limited risk for human health, it is necessary to improve general education to reduce canine faecal pollution towards a widespread awareness of health risks.

Cryptosporidium spp. in pet birds: genetic diversity and potential public health significance.

PubMed

Qi, Meng; Wang, Rongjun; Ning, Changshen; Li, Xiaoyu; Zhang, Longxian; Jian, Fuchun; Sun, Yanru; Xiao, Lihua

2011-08-01

To characterize the prevalence and assess the zoonotic transmission burden of Cryptosporidium species/genotypes in pet birds in Henan, China, 434 fecal samples were acquired from 14 families of birds in pet shops. The overall prevalence of Cryptopsoridium was 8.1% (35/434) by the Sheather's sugar flotation technique. The Cryptosporidium-positive samples were analyzed by DNA sequence analysis of the small subunit (SSU) rRNA gene. Three Cryptosporidium species and two genotypes were identified, including C. baileyi (18/35 or 51.4%) in five red-billed leiothrixes (Leiothrix lutea), four white Java sparrows (Padda oryzivora), four common mynas (Acridotheres tristis), two zebra finches (Taeniopygia guttata), a crested Lark (Galerida cristata), a Gouldian finch (Chloebia gouldiae), and a black-billed magpie (Pica pica); Cryptosporidium meleagridis (3/35 or 8.6%) in a Bohemian waxwing (Bombycilla garrulus), a Rufous turtle dove (Streptopelia orientalis), and a fan-tailed pigeon (Columba livia); Cryptosporidium galli (5/35 or 14.3%) in four Bohemian waxwings (Bombycilla garrulus) and a silver-eared Mesia (Leiothrix argentauris); Cryptosporidium avian genotype III (3/35 or 8.6%) in two cockatiels (Nymphicus hollandicus) and a red-billed blue magpie (Urocissa erythrorhyncha); and Cryptosporidium avian genotype V (6/35 or 17.1%) in six cockatiels (Nymphicus hollandicus). Among the pet birds, 12 species represented new hosts for Cryptosporidum infections. The presence of C. meleagridis raises questions on potential zoonotic transmission of cryptosporidiosis from pet birds to humans. Copyright © 2011 Elsevier Inc. All rights reserved.

CRYPTOSPORIDIUM AND GIARDIA IN STORMWATER AS A THREAT TO DRINKING WATER SUPPLIES

EPA Science Inventory

Since the first identified Cryptosporidium outbreak in the United Kingdom in 1983, the pathogens Cryptosporidium and Giardia have become the subject of growing local, state, and national concern. In the last decade, these organisms have been the causative agent of many gastroint...

EVALUATING CRYPTOSPORIDIUM AND GIARDIA IN STORMWATER AS A THREAT TO DRINKING WATER SUPPLIES

EPA Science Inventory

Since the first identified Cryptosporidium outbreak in the United Kingdom in 1983, the
pathogens Cryptosporidium and Giardia have become the subject of growing local, state, and
national concern. Both organisms have been the causative agent of many gastrointestinal
illn...

Occurrence of waterborne pathogens and Escherichia coli at offshore drinking water intakes in lake Ontario.

PubMed

Edge, T A; Khan, I U H; Bouchard, R; Guo, J; Hill, S; Locas, A; Moore, L; Neumann, N; Nowak, E; Payment, P; Yang, R; Yerubandi, R; Watson, S

2013-10-01

The occurrence of waterborne pathogens was investigated at three drinking water intakes located about 2 km offshore in Lake Ontario. Water sampling was conducted over 3 years for Campylobacter spp., Cryptosporidium spp., Giardia spp., cultivable enteric viruses, and water quality parameters. All pathogens were detected in the offshore source water for each water treatment plant (WTP1 to WTP3), although at relatively low frequencies and concentrations. Giardia was the most common pathogen, occurring in 36% of water samples from the influent of WTP1 (n = 46), and with a maximum concentration of 0.70 cysts/liter in this influent. Cryptosporidium occurred as frequently as 15% in the WTP2 influent (n = 35), with a maximum concentration of 0.40 oocysts/liter in the WTP1 influent. The human Bacteroidales HF183 DNA marker was most common in the WTP1 influent (19%), and this was the only WTP where the Cryptosporidium hominis genotype was detected. No water quality parameter was predictive of pathogen occurrence across all three WTP influents. Escherichia coli was often below detection when pathogens were detected, and spikes in E. coli concentrations often did not coincide with pathogen occurrence. After summer rain events, river plumes had E. coli concentrations as high as 222 CFU/100 ml in surface waters 2 km offshore, without impacting drinking water intakes below the thermocline on the lake bottom. At times, prechlorination to control mussels at offshore intake cribs compromised the use of E. coli for "raw" water quality assessment, particularly for chlorine-resistant Cryptosporidium. E. coli measured by standard methods did not reliably predict pathogen occurrence at drinking water intakes in offshore ecosystems.

Immunomagnetic Separation of Cryptosporidium parvum from Source Water Samples of Various Turbidities

PubMed Central

Bukhari, Z.; McCuin, R. M.; Fricker, C. R.; Clancy, J. L.

1998-01-01

Immunomagnetic separation (IMS) procedures which specifically capture Cryptosporidium oocysts and have the potential to isolate oocysts from debris have become commercially available. We compared two IMS kits (kit DB [Dynabeads anti-Cryptosporidium; product no. 730.01; Dynal A.S., Oslo, Norway] and kit IC1 [Crypto Scan IMS; product no. R10; Clearwater Diagnostics Company, LLC, Portland, Maine]) and a modification of kit IC1 (kit IC2 [Crypto Scan IMS; product no. R10; Clearwater Diagnostics Company, LLC]) at three turbidity levels (50, 500, and 5,000 nephelometric turbidity units [ntu]) by using water matrices obtained from different geographical locations. In deionized water, kit DB yielded recoveries between 68 and 83%, whereas the recoveries obtained with kits IC1 and IC2 were more variable and ranged from 0.2 to 74.5%. In water matrices with turbidity levels up to 500 ntu, the oocyst recoveries were more variable with kit DB; however, the recoveries were similar to those obtained in deionized water. In contrast, there were notable reductions in oocyst recoveries in the turbid matrices with kits IC1 and IC2, and the highest recovery (8.3%) was obtained with a 50-ntu sample. An examination of the effects of age on oocyst recovery with kit DB revealed that oocysts up to 16 weeks old yielded recoveries similar to the recoveries observed with fresh oocysts. These data indicate that all IMS kits do not perform equally well, and it is important to conduct in-house quality assurance work before a commercially available IMS kit is selected to replace flotation procedures for recovery of Cryptosporidium oocysts. PMID:9797313

Occurrence of Waterborne Pathogens and Escherichia coli at Offshore Drinking Water Intakes in Lake Ontario

PubMed Central

Khan, I. U. H.; Bouchard, R.; Guo, J.; Hill, S.; Locas, A.; Moore, L.; Neumann, N.; Nowak, E.; Payment, P.; Yang, R.; Yerubandi, R.; Watson, S.

2013-01-01

The occurrence of waterborne pathogens was investigated at three drinking water intakes located about 2 km offshore in Lake Ontario. Water sampling was conducted over 3 years for Campylobacter spp., Cryptosporidium spp., Giardia spp., cultivable enteric viruses, and water quality parameters. All pathogens were detected in the offshore source water for each water treatment plant (WTP1 to WTP3), although at relatively low frequencies and concentrations. Giardia was the most common pathogen, occurring in 36% of water samples from the influent of WTP1 (n = 46), and with a maximum concentration of 0.70 cysts/liter in this influent. Cryptosporidium occurred as frequently as 15% in the WTP2 influent (n = 35), with a maximum concentration of 0.40 oocysts/liter in the WTP1 influent. The human Bacteroidales HF183 DNA marker was most common in the WTP1 influent (19%), and this was the only WTP where the Cryptosporidium hominis genotype was detected. No water quality parameter was predictive of pathogen occurrence across all three WTP influents. Escherichia coli was often below detection when pathogens were detected, and spikes in E. coli concentrations often did not coincide with pathogen occurrence. After summer rain events, river plumes had E. coli concentrations as high as 222 CFU/100 ml in surface waters 2 km offshore, without impacting drinking water intakes below the thermocline on the lake bottom. At times, prechlorination to control mussels at offshore intake cribs compromised the use of E. coli for “raw” water quality assessment, particularly for chlorine-resistant Cryptosporidium. E. coli measured by standard methods did not reliably predict pathogen occurrence at drinking water intakes in offshore ecosystems. PMID:23835181

Water-Quality Changes Caused by Riverbank Filtration Between the Missouri River and Three Pumping Wells of the Independence, Missouri, Well Field 2003-05

USGS Publications Warehouse

Kelly, Brian P.; Rydlund, Jr., Paul H.

2006-01-01

Riverbank filtration substantially improves the source-water quality of the Independence, Missouri well field. Coliform bacteria, Cryptosporidium, Giardia, viruses and selected constituents were analyzed in water samples from the Missouri River, two vertical wells, and a collector well. Total coliform bacteria, Cryptosporidium, Giardia, and total culturable viruses were detected in the Missouri River, but were undetected in samples from wells. Using minimum reporting levels for non-detections in well samples, minimum log removals were 4.57 for total coliform bacteria, 1.67 for Cryptosporidium, 1.67 for Giardia, and 1.15 for total culturable virus. Ground-water flow rates between the Missouri River and wells were calculated from water temperature profiles and ranged between 1.2 and 6.7 feet per day. Log removals based on sample pairs separated by the traveltime between the Missouri River and wells were infinite for total coliform bacteria (minimum detection level equal to zero), between 0.8 and 3.5 for turbidity, between 1.5 and 2.1 for Giardia, and between 0.4 and 2.6 for total culturable viruses. Cryptosporidium was detected once in the Missouri River but no corresponding well samples were available. No clear relation was evident between changes in water quality in the Missouri River and in wells for almost all constituents. Results of analyses for organic wastewater compounds and the distribution of dissolved oxygen, specific conductance, and temperature in the Missouri River indicate water quality on the south side of the river was moderately influenced by the south bank inflows to the river upstream from the Independence well field.

Occurrence and removal of Giardia spp. cysts and Cryptosporidium spp. oocysts from a municipal wastewater treatment plant in Brazil.

PubMed

Santos, Priscila Ribeiro Dos; Daniel, Luiz Antonio

2017-05-01

Sewage and sewage sludge have been recognized as potential sources of two important waterborne pathogenic protozoa: Giardia spp. and Cryptosporidium spp. Due to the lack of studies about the occurrence of these pathogens in sewage and sludge in Brazil, an investigation was conducted at various stages of a municipal wastewater treatment plant (WWTP) aiming to assess the occurrence of Giardia spp. cysts and Cryptosporidium spp. oocysts, their removal by the treatment processes, which are upflow anaerobic sludge blanket (UASB) reactor and dissolved air flotation process, and also the correlations between protozoa and indicator microorganisms. Significant quantities of cysts were detected in 100% of the analyzed wastewater samples, while oocysts were detected only in 39.0% of all wastewater samples. The overall removal of Giardia spp. cysts from the WWTP was on average 2.03 log, and the UASB reactor was more efficient than flotation. The sludge samples presented high quantities of (oo)cysts, implying the risks of contamination in the case of sludge reuse or inadequate disposal. Giardiasis prevalence was estimated between 2.21% and 6.7% for the population served by the WWTP, while cryptosporidiosis prevalence was much lower. Significant positive correlation was obtained only between cysts and Clostridium spores in anaerobic effluent.

Molecular Detection of the Carriage Rate of Four Intestinal Protozoa with Real-Time Polymerase Chain Reaction: Possible Overdiagnosis of Entamoeba histolytica in Nigeria

PubMed Central

Efunshile, Michael A.; Ngwu, Bethrand A. F.; Kurtzhals, Jørgen A. L.; Sahar, Sumrin; König, Brigitte; Stensvold, Christen R.

2015-01-01

Diarrhea remains the second largest killer of children worldwide, and Nigeria ranks number two on the list of global deaths attributable to diarrhea. Meanwhile, prevalence studies on potentially diarrheagenic protozoa in asymptomatic carriers using molecular detection methods remain scarce in sub-Saharan countries. To overcome sensitivity issues related to microscopic detection and identification of cysts in stool concentrates, real-time polymerase chain reaction (PCR) was used to analyze genomic DNAs extracted from stool samples from 199 healthy school children for Entamoeba histolytica, E. dispar, Giardia intestinalis, and Cryptosporidium. Questionnaires were administered for epidemiological data collection. E. histolytica was not detected in any of the samples, whereas Giardia (37.2%), E. dispar (18.6%), and Cryptosporidium (1%) were found. Most of the children sourced their drinking water from community wells (91%), while the majority disposed of feces in the bush (81.9%). Our study is the first to use real-time PCR to evaluate the epidemiology of E. histolytica, Giardia, and Cryptosporidium in Nigeria where previous studies using traditional diagnostic techniques have suggested higher and lower carriage rates of E. histolytica and Giardia, respectively. It is also the first study to accurately identify the prevalence of common potentially diarrheagenic protozoa in asymptomatic carriers in sub-Saharan Africa. PMID:26101274

Spinacia oleracea L. leaf stomata harboring Cryptosporidium parvum oocysts: A potential threat for food safety

USDA-ARS?s Scientific Manuscript database

Scientific literature documents the prevalence of Cryptosporidium oocysts in irrigation waters and on fresh produce. In the present study spinach leaves were experimentally exposed to Cryptosporidium oocysts which were subsequently irrigated with clean water daily for 5 days. As determined by confoc...

Using Propidium Monoazide as an Enrichment Step to Remove Extraneous DMA and Non-Viable Organisms for Cryptosporidium Genotyping

EPA Science Inventory

Cryptosporidium is an important protozoan parasite that continues to cause waterborne disease outbreaks worldwide. Current methods to monitor for Cryptosporidium oocysts in water are microscopy-based USEPA Methods 1622 and 1623. These methods assess total levels of oocysts in s...

Cryptosporidium Off-The-Slide Genotyping: Recovery of Oocysts Extracted from Slides Examined by Method 1623

EPA Science Inventory

Cryptosporidium has been found world wide and is an important waterborne parasite causing a self-limiting diarrhea; however, in the immunocompromised it may become chronic and can even lead to death. To characterize the risk of exposure to Cryptosporidium in water the USEPA enac...

The use of an atmospheric cold plasma jet to inactivate Cryptosporidium parvum oocysts on cilantro

USDA-ARS?s Scientific Manuscript database

Introduction: In 2015, the CDC reported a rise in outbreaks linked to parasites like Cryptosporidium. Outbreaks of Cryptosporidium parvum have been associated with contaminated drinking or recreational water; however, there is growing concern that oocysts may become a more common contaminant in food...

REMOVAL OF CRYPTOSPORIDIUM BY IN-LINE FILTRATION AS A FUNCTION OF OOCYST AGE AND PRESERVATION METHOD

EPA Science Inventory

This study examined the impacts of oocyst preservation method and age on the removal of seeded Cryptosporidium oocysts by in-line filtration. An existing study has investigated the infectivity of Cryptosporidium parvum as a function of preservation method and oocyst age. Simila...

REMOVAL OF CRYPTOSPORIDIUM BY IN-LINE FILTRATION AS A FUNCTION OF OOCYST AGE AND PRESERVATION METHOD

EPA Science Inventory

This study examined the impacts of oocyst preservation method and age on the removal of seeded Cryptosporidium oocysts by in-line filtration. An existing study has investigated the infectivity of Cryptosporidium Parvum as a function of preservation method and oocyst age. Simila...

EPA ORD/REGION 2 COOPERATIVE EFFORT: EVALUATING CRYPTOSPORIDIUM AND GIARDIA IN URBAN STORMWATER

EPA Science Inventory

Since the first identified Cryptosporidium outbreak in the United Kingdom in 1983, the pathogens Cryptosporidium and Giardia have become the subject of growing local, state, and national concern. Both organisms have been the causative agent of many gas...

INVESTIGATION AND EVALUATION OF CRYPTOSPORIDIUM AND GIARDIA IN COMBINED SEWER OVERFLOW (CSO) AND STORMWATER RUNOFF

EPA Science Inventory

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