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Sample records for cryptosporidium parvum infections

  1. Quantitative-PCR Assessment of Cryptosporidium parvum Cell Culture Infection

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    Di Giovanni, George D.; LeChevallier, Mark W.

    2005-01-01

    A quantitative TaqMan PCR method was developed for assessing the Cryptosporidium parvum infection of in vitro cultivated human ileocecal adenocarcinoma (HCT-8) cell cultures. This method, termed cell culture quantitative sequence detection (CC-QSD), has numerous applications, several of which are presented. CC-QSD was used to investigate parasite infection in cell culture over time, the effects of oocyst treatment on infectivity and infectivity assessment of different C. parvum isolates. CC-Q...

  2. [The ecology of Cryptosporidium parvum infection in small rodent populations].

    Science.gov (United States)

    Bajer, A; Bednarska, M; Siński, E

    2001-01-01

    The prevalence and abundance of Cryptosporidium parvum were studied over a three year period (1997-1999) in three species of rodents sampled from forest and abandoned fields in the Mazury Lake District, Poland. The overall prevalence was consistently higher in voles compared with Apodemus flavicollis (70.6% in Clethrionomys glareolus, 73.0% in Microtus arvalis and 27.8% in A. flavicollis). The prevalence and abundance of infection also varied across the 3 years of the study with 1998 being the year of higher prevalence and abundance of the parasite. Fewer older animals carried the infection, and their infections were relatively mild. We found no consistent pattern of seasonal changes despite the significance of seasonal differences. Host sex did not influence either the prevalence or abundance of infection with C. parvum. A great proportion of recaptured voles developed chronic infections between consecutive trapping sessions and only a small number of animals recovered. However, yellow-necked mice seem to be much more resistant to infection that became self-limiting. Our results firmly establish that the common woodland and grassland wild rodents in the Mazury Lake District constitute a significant and hazardous reservoir of C. parvum for animals and humans.

  3. Pathogenicity of Cryptosporidium parvum - evaluation of an animal infection model

    DEFF Research Database (Denmark)

    Enemark, Heidi L.; Bille-Hansen, Vivi; Lind, Peter

    2003-01-01

    With the intention of developing a standardised method for assessment of pathogenicity of Cryptosporidium parvum, the CPB-0 isolate was studied by propagation in 1-day-old calves followed by inoculation into specific pathogen free (SPF) piglets. The experiment was repeated. Diarrhoea and shedding...

  4. A new in vitro model using small intestinal epithelial cells to enhance infection of Cryptosporidium parvum

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    To better understand and study the infection of the protozoan parasite Cryptosporidium parvum, a more sensitive in vitro assay is required. In vivo, this parasite infects the epithelial cells of the microvilli layer in the small intestine. While cell infection models using colon,...

  5. INTESTINAL AND PULMONARY INFECTION BY Cryptosporidium parvum IN TWO PATIENTS WITH HIV/AIDS

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    Fábio Tadeu Rodrigues REINA

    2016-01-01

    Full Text Available We describe two patients with HIV/AIDS who presented pulmonary and intestinal infection caused by Cryptosporidium parvum, with a fatal outcome. The lack of available description of changes in clinical signs and radiographic characteristics of this disease when it is located in the extra-intestinal region causes low prevalence of early diagnosis and a subsequent lack of treatment.

  6. INTESTINAL AND PULMONARY INFECTION BY Cryptosporidium parvum IN TWO PATIENTS WITH HIV/AIDS

    Science.gov (United States)

    REINA, Fábio Tadeu Rodrigues; RIBEIRO, Camila Aparecida; de ARAÚJO, Ronalda Silva; MATTÉ, Maria Helena; CASTANHO, Roberto Esteves Pires; TANAKA, Ioshie Ibara; VIGGIANI, Ana Maria Ferreira Sornas; MARTINS, Luciamáre Perinetti Alves

    2016-01-01

    We describe two patients with HIV/AIDS who presented pulmonary and intestinal infection caused by Cryptosporidium parvum, with a fatal outcome. The lack of available description of changes in clinical signs and radiographic characteristics of this disease when it is located in the extra-intestinal region causes low prevalence of early diagnosis and a subsequent lack of treatment. PMID:27007564

  7. Natural infection with zoonotic subtype of Cryptosporidium parvum in Capybara (Hydrochoerus hydrochaeris) from Brazil.

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    Meireles, Marcelo Vasconcelos; Soares, Rodrigo Martins; Bonello, Fábio; Gennari, Solange Maria

    2007-06-20

    A total of 145 capybara (Hydrochoerus hydrochaeris) fecal samples from the state of São Paulo, Brazil, were screened for Cryptosporidium spp. oocysts using the malachite green method. Eight samples (5.52%) showed positive results and were further submitted to nested PCR reaction for amplification of fragments of 18S rRNA gene and 60-kDa glycoprotein gene for determination of species, alleles and subtypes of Cryptosporidium. Sequencing of the PCR products of the 18S rRNA gene fragments and 60-kDa glycoprotein gene fragments showed that for both genes all Cryptosporidium isolates from capybara were respectively 100% genetically similar to a bovine isolate of C. parvum and to C. parvum subtype IIaA15G2R1. To the best of our knowledge this is the first report of Cryptosporidium infection in this rodent. The finding of zoonotic C. parvum infection in a semi-aquatic mammal that inhabits anthroponotic habitats raises the concern that human water supplies may be contaminated with zoonotic Cryptosporidium oocysts from wildlife.

  8. Effects of hydrogen peroxide on infections of cryptosporidium parvum in vitro

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Effects of hydrogen peroxide (H2O2)on Cryptosporidium parvum infection in vitro were studied in this paper, using an oocyst excystation assay, a cell culture model, and a free radical inhibition technique. H2O2 treatment at 500 and 1 000 μmol/L significantly inhibited excystation of bleach-treated oocysts (P<0.01). Concentrations of H2O2 at 500 and 750 μmol/L resulted in a significant decrease in C. Parvum infection at 35.77% and 58.16% respectively, when compared with the untreated control at 48 hours postinoculation. Surprisingly, C. parvum infection were significantly increased by 22.21% to 39.33% following treatment with 50 (P<0.01), 100 (P<0.01) or 200 (P<0.05) μmol/L H2O2, respectively. Stimulatory effect with treatment of 100 μmol/L H2O2 was most obvious, compared with the untreated control at 48 hours postinoculation. Effects of H2O2 on C. parvum at 24, 48 and 96 hours postinoculation were similar, the highest infection being infection at 48 hours postinoculation, with the maximum inhibitory effect being seen at 96 hours postinoculation. The stimulatory and inhibitory effects of H2O2 treatment on C. parvum infection were, to a certain extent, abolished in the presence of free radical scavengers, reduced glutathione or mannitol. These observations indicate that reactive oxygen species (ROS), such as H2O2, may play a role in the course of C. parvum infection. This is the first time to demonstrate a significant action of ROS in C. parvum infection in vitro.

  9. [The co-occurence of Cryptosporidium parvum, Giardia spp. and helminth infections in small rodent populations].

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    Bajer, Anna; Behnke, Jerzy M; Bednarska, Małgorzata; Kuliś, Karolina; Siński, Edward

    2004-01-01

    During long-term (1998-2000) studies on rodent parasite populations in Mazury lake district there were collected and analyzed data on co-occurrence of intestinal protozoa (Cryptosporidium parvum, Giardia spp.) and helminths. There were performed 178 autopsies of common vole Microtus arvalis, 85 autopsies of yellow-necked mouse Apodemus flavicollis and 386 autopsies of bank vole Clethrionomys glareolus. Positive effect of helminth infections was found in C. glareolus. Voles infected with nematode Heligmosomum mixtum showed higher prevalence of C. parvum and Giardia spp. than voles infected with Heligmosomoides glareoli. The host age took part in these interactions and positive effect of co-occurrence was mainly observed in voles older than 3 months. The other intrinsic (host sex) or extrinsic (season and year of study) factors influenced interactions between parasites. Presented results revealed that helminth infections may facilitate chronic infections of intestinal protozoa in rodent populations.

  10. Cryptosporidium parvum: infectivity and pathogenicity of the 'porcine' genotype

    DEFF Research Database (Denmark)

    Enemark, Heidi L.; Ahrens, Peter; Bille-Hansen, Vivi

    2003-01-01

    mild clinical signs in piglets despite the excretion of high numbers of oocysts. Concomitant infection with rotavirus, however, caused a dramatic aggravation of the clinical signs, and 5 of 6 experimentally infected piglets died. CPP-13 appeared to be adapted to porcine hosts as illustrated by the lack...

  11. Comparison of in vitro cell culture and a mouse assay for measuring infectivity of Cryptosporidium parvum.

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    Rochelle, Paul A; Marshall, Marilyn M; Mead, Jan R; Johnson, Anne M; Korich, Dick G; Rosen, Jeffrey S; De Leon, Ricardo

    2002-08-01

    In vitro cell cultures were compared to neonatal mice for measuring the infectivity of five genotype 2 isolates of Cryptosporidium parvum. Oocyst doses were enumerated by flow cytometry and delivered to animals and cell monolayers by using standardized procedures. Each dose of oocysts was inoculated into up to nine replicates of 9 to 12 mice or 6 to 10 cell culture wells. Infections were detected by hematoxylin and eosin staining in CD-1 mice, by reverse transcriptase PCR in HCT-8 and Caco-2 cells, and by immunofluorescence microscopy in Madin-Darby canine kidney (MDCK) cells. Infectivity was expressed as a logistic transformation of the proportion of animals or cell culture wells that developed infection at each dose. In most instances, the slopes of the dose-response curves were not significantly different when we compared the infectivity models for each isolate. The 50% infective doses for the different isolates varied depending on the method of calculation but were in the range from 16 to 347 oocysts for CD-1 mice and in the ranges from 27 to 106, 31 to 629, and 13 to 18 oocysts for HCT-8, Caco-2, and MDCK cells, respectively. The average standard deviations for the percentages of infectivity for all replicates of all isolates were 13.9, 11.5, 13.2, and 10.7% for CD-1 mice, HCT-8 cells, Caco-2 cells, and MDCK cells, respectively, demonstrating that the levels of variability were similar in all assays. There was a good correlation between the average infectivity for HCT-8 cells and the results for CD-1 mice across all isolates for untreated oocysts (r = 0.85, n = 25) and for oocysts exposed to ozone and UV light (r = 0.89, n = 29). This study demonstrated that in vitro cell culture was equivalent to the "gold standard," mouse infectivity, for measuring the infectivity of C. parvum and should therefore be considered a practical and accurate alternative for assessing oocyst infectivity and inactivation. However, the high levels of variability displayed by all

  12. Direct ELISA aided coprological diagnosis of Cryptosporidium parvum infection in diarrheic neonatal calves in Mosul city, Iraq

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    Israa Al-Robaiee

    2014-03-01

    Full Text Available This study was conducted for the detection of Cryptosporidium (C. parvum infection in neonatal calves. A total of 220 fecal samples (diarrheic 110 and nondiarrheic 110 of neonatal calves were collected from Mosul city, Iraq over a period of 16 months from November 2010 to March 2012. The age of the calves ranged from 1 to 30 days. All the fecal samples were analyzed by capture direct ELISA. The infection was found in 29.0% (n=32/110 of the diarrheic calves. The infection was mostly prevalent (p<0.001 in the calves of three weeks of age. No C. parvum infection could be detected in the nondiarrheic animals.

  13. Pilot-Scale Pulsed UV Light Irradiation of Experimentally Infected Raspberries Suppresses Cryptosporidium parvum Infectivity in Immunocompetent Suckling Mice.

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    Le Goff, L; Hubert, B; Favennec, L; Villena, I; Ballet, J J; Agoulon, A; Orange, N; Gargala, G

    2015-12-01

    Cryptosporidium spp., a significant cause of foodborne infection, have been shown to be resistant to most chemical food disinfectant agents and infective for weeks in irrigation waters and stored fresh vegetal produce. Pulsed UV light (PL) has the potential to inactivate Cryptosporidium spp. on surfaces of raw or minimally processed foods or both. The present study aimed to evaluate the efficacy of PL on viability and in vivo infectivity of Cryptosporidium parvum oocysts present on raspberries, a known source of transmission to humans of oocyst-forming apicomplexan pathogens. The skin of each of 20 raspberries was experimentally inoculated with five 10-μl spots of an oocyst suspension containing 6 × 10(7) oocysts per ml (Nouzilly isolate). Raspberries were irradiated by PL flashes (4 J/cm(2) of total fluence). This dose did not affect colorimetric or organoleptic characteristics of fruits. After immunomagnetic separation from raspberries, oocysts were bleached and administered orally to neonatal suckling mice. Seven days after infection, mice were euthanized, and the number of oocysts in the entire small intestine was individually assessed by immunofluorescence flow cytometry. Three of 12 and 12 of 12 inoculated mice that received 10 and 100 oocysts isolated from nonirradiated raspberries, respectively, were found infected. Four of 12 and 2 of 12 inoculated mice that received 10(3) and 10(4) oocysts from irradiated raspberries, respectively, were found infected. Oocyst counts were lower in animals inoculated with 10(3) and 10(4) oocysts from irradiated raspberries (92 ± 144 and 38 ± 82, respectively) than in animals infected with 100 oocysts from nonirradiated raspberries (35,785 ± 66,221, P = 0.008). PL irradiation achieved oocyst reductions of 2 and 3 log for an inoculum of 10(3) and 10(4) oocysts, respectively. The present pilot-scale evaluation suggests that PL is an effective mode of decontamination for raspberries and prompts further applicability

  14. Temporal and spatial dynamics of Cryptosporidium parvum infection on dairy farms in the New York City Watershed: a cluster analysis based on crude and Bayesian risk estimates

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    Mohammed Hussni O

    2010-06-01

    Full Text Available Abstract Background Cryptosporidium parvum is one of the most important biological contaminants in drinking water that produces life threatening infection in people with compromised immune systems. Dairy calves are thought to be the primary source of C. parvum contamination in watersheds. Understanding the spatial and temporal variation in the risk of C. parvum infection in dairy cattle is essential for designing cost-effective watershed management strategies to protect drinking water sources. Crude and Bayesian seasonal risk estimates for Cryptosporidium in dairy calves were used to investigate the spatio-temporal dynamics of C. parvum infection on dairy farms in the New York City watershed. Results Both global (Global Moran's I and specific (SaTScan cluster analysis methods revealed a significant (p C. parvum infection in all herds in the summer (p = 0.002, compared to the rest of the year. Bayesian estimates did not show significant spatial autocorrelation in any season. Conclusions Although we were not able to identify seasonal clusters using Bayesian approach, crude estimates highlighted both temporal and spatial clusters of C. parvum infection in dairy herds in a major watershed. We recommend that further studies focus on the factors that may lead to the presence of C. parvum clusters within the watershed, so that monitoring and prevention practices such as stream monitoring, riparian buffers, fencing and manure management can be prioritized and improved, to protect drinking water supplies and public health.

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

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

    2014-10-01

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

  16. NF-kappaB p65-dependent transactivation of miRNA genes following Cryptosporidium parvum infection stimulates epithelial cell immune responses.

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    Zhou, Rui; Hu, Guoku; Liu, Jun; Gong, Ai-Yu; Drescher, Kristen M; Chen, Xian-Ming

    2009-12-01

    Cryptosporidium parvum is a protozoan parasite that infects the gastrointestinal epithelium and causes diarrheal disease worldwide. Innate epithelial immune responses are key mediators of the host's defense to C. parvum. MicroRNAs (miRNAs) regulate gene expression at the posttranscriptional level and are involved in regulation of both innate and adaptive immune responses. Using an in vitro model of human cryptosporidiosis, we analyzed C. parvum-induced miRNA expression in biliary epithelial cells (i.e., cholangiocytes). Our results demonstrated differential alterations in the mature miRNA expression profile in cholangiocytes following C. parvum infection or lipopolysaccharide stimulation. Database analysis of C. parvum-upregulated miRNAs revealed potential NF-kappaB binding sites in the promoter elements of a subset of miRNA genes. We demonstrated that mir-125b-1, mir-21, mir-30b, and mir-23b-27b-24-1 cluster genes were transactivated through promoter binding of the NF-kappaB p65 subunit following C. parvum infection. In contrast, C. parvum transactivated mir-30c and mir-16 genes in cholangiocytes in a p65-independent manner. Importantly, functional inhibition of selected p65-dependent miRNAs in cholangiocytes increased C. parvum burden. Thus, we have identified a panel of miRNAs regulated through promoter binding of the NF-kappaB p65 subunit in human cholangiocytes in response to C. parvum infection, a process that may be relevant to the regulation of epithelial anti-microbial defense in general.

  17. NF-kappaB p65-dependent transactivation of miRNA genes following Cryptosporidium parvum infection stimulates epithelial cell immune responses.

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    Rui Zhou

    2009-12-01

    Full Text Available Cryptosporidium parvum is a protozoan parasite that infects the gastrointestinal epithelium and causes diarrheal disease worldwide. Innate epithelial immune responses are key mediators of the host's defense to C. parvum. MicroRNAs (miRNAs regulate gene expression at the posttranscriptional level and are involved in regulation of both innate and adaptive immune responses. Using an in vitro model of human cryptosporidiosis, we analyzed C. parvum-induced miRNA expression in biliary epithelial cells (i.e., cholangiocytes. Our results demonstrated differential alterations in the mature miRNA expression profile in cholangiocytes following C. parvum infection or lipopolysaccharide stimulation. Database analysis of C. parvum-upregulated miRNAs revealed potential NF-kappaB binding sites in the promoter elements of a subset of miRNA genes. We demonstrated that mir-125b-1, mir-21, mir-30b, and mir-23b-27b-24-1 cluster genes were transactivated through promoter binding of the NF-kappaB p65 subunit following C. parvum infection. In contrast, C. parvum transactivated mir-30c and mir-16 genes in cholangiocytes in a p65-independent manner. Importantly, functional inhibition of selected p65-dependent miRNAs in cholangiocytes increased C. parvum burden. Thus, we have identified a panel of miRNAs regulated through promoter binding of the NF-kappaB p65 subunit in human cholangiocytes in response to C. parvum infection, a process that may be relevant to the regulation of epithelial anti-microbial defense in general.

  18. Cryptosporidium parvum infection in SCID mice infected with only one oocyst: qPCR assessment of parasite replication in tissues and development of digestive cancer.

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    Sadia Benamrouz

    Full Text Available Dexamethasone (Dex treated Severe Combined Immunodeficiency (SCID mice were previously described as developing digestive adenocarcinoma after massive infection with Cryptosporidium parvum as soon as 45 days post-infection (P.I.. We aimed to determine the minimum number of oocysts capable of inducing infection and thereby gastrointestinal tumors in this model. Mice were challenged with calibrated oocyst suspensions containing intended doses of: 1, 10, 100 or 10(5 oocysts of C. parvum Iowa strain. All administered doses were infective for animals but increasing the oocyst challenge lead to an increase in mice infectivity (P = 0.01. Oocyst shedding was detected at 7 days P.I. after inoculation with more than 10 oocysts, and after 15 days in mice challenged with one oocyst. In groups challenged with lower inocula, parasite growth phase was significantly higher (P = 0.005 compared to mice inoculated with higher doses. After 45 days P.I. all groups of mice had a mean of oocyst shedding superior to 10,000 oocyst/g of feces. The most impressive observation of this study was the demonstration that C. parvum-induced digestive adenocarcinoma could be caused by infection with low doses of Cryptosporidium, even with only one oocyst: in mice inoculated with low doses, neoplastic lesions were detected as early as 45 days P.I. both in the stomach and ileo-caecal region, and these lesions could evolve in an invasive adenocarcinoma. These findings show a great amplification effect of parasites in mouse tissues after challenge with low doses as confirmed by quantitative PCR. The ability of C. parvum to infect mice with one oocyst and to develop digestive adenocarcinoma suggests that other mammalian species including humans could be also susceptible to this process, especially when they are severely immunocompromised.

  19. Cryptosporidium parvum, a potential cause of colic adenocarcinoma

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    Pinon Anthony

    2007-11-01

    Full Text Available Abstract Background Cryptosporidiosis represents a major public health problem. This infection has been reported worldwide as a frequent cause of diarrhoea. Particularly, it remains a clinically significant opportunistic infection among immunocompromised patients, causing potentially life-threatening diarrhoea in HIV-infected persons. However, the understanding about different aspects of this infection such as invasion, transmission and pathogenesis is problematic. Additionally, it has been difficult to find suitable animal models for propagation of this parasite. Efforts are needed to develop reproducible animal models allowing both the routine passage of different species and approaching unclear aspects of Cryptosporidium infection, especially in the pathophysiology field. Results We developed a model using adult severe combined immunodeficiency (SCID mice inoculated with Cryptosporidium parvum or Cryptosporidium muris while treated or not with Dexamethasone (Dex in order to investigate divergences in prepatent period, oocyst shedding or clinical and histopathological manifestations. C. muris-infected mice showed high levels of oocysts excretion, whatever the chemical immunosuppression status. Pre-patent periods were 11 days and 9.7 days in average in Dex treated and untreated mice, respectively. Parasite infection was restricted to the stomach, and had a clear preferential colonization for fundic area in both groups. Among C. parvum-infected mice, Dex-treated SCID mice became chronic shedders with a prepatent period of 6.2 days in average. C. parvum-inoculated mice treated with Dex developed glandular cystic polyps with areas of intraepithelial neoplasia, and also with the presence of intramucosal adenocarcinoma. Conclusion For the first time C. parvum is associated with the formation of polyps and adenocarcinoma lesions in the gut of Dex-treated SCID mice. Additionally, we have developed a model to compare chronic muris and parvum

  20. Does the use of tubular digesters to treat livestock waste lower the risk of infection from Cryptosporidium parvum and Giardia lamblia?

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    Kinyua, Maureen N; Wald, Ileana; Camacho-Céspedes, Fabricio; Izurieta, Ricardo; Haas, Charles N; Ergas, Sarina J

    2016-10-01

    Worldwide, high incidences of cryptosporidiosis and giardiasis are attributed to livestock waste. Quantitative microbial risk assessment can be used to estimate the risk of livestock related infections from Cryptosporidium parvum and Giardia lamblia. The objective of this paper was to assess the occupational and public health risks associated with management of raw and anaerobically digested livestock waste in two rural communities in Costa Rica based on fomite, soil and crop contamination and livestock waste management exposure pathways. Risks related to cattle waste were greater than swine waste due to cattle shedding more (oo)cysts. Cryptosporidium parvum also posed a greater risk than Giardia lamblia in all exposure pathways due to livestock shedding high loads of Cryptosporidium parvum oocysts and oocysts' lower inactivation rates during anaerobic digestion compared with Giardia lamblia cysts. The risk of infection from exposure to contaminated soil and crops was significantly lower for a community using tubular anaerobic digesters to treat livestock waste compared to a community where the untreated waste was applied to soil. The results indicate that treatment of livestock waste in small-scale tubular anaerobic digesters has the potential to significantly decrease the risk of infection below the World Health Organization's acceptable individual annual risk of infection (10(-4)).

  1. AGING OF CRYPTOSPORIDIUM PARVUM OOCYSTS STUDIED BY MALDI-TF MS

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    Cryptosporidium parvum is an obligate protozoan parasite found in surface waters. It is the etiological agent for cryptosporidiosis, a parasitic infection that causes severe gastrointestinal illness which is potentially fatal among immuno-compromised individuals. This water borne...

  2. MOLECULAR CLONING AND ANALYSIS OF THE CRYPTOSPORIDIUM PARVUM AMINOPEPTIDASE N GENE. (R829180)

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    Cryptosporidium parvum proteases have been associated with release of infective sporozoites from oocysts, and their specific inhibition blocks parasite excystation in vitro. Additionally, proteases have been implicated in the processing of parasite adhesion molecules fo...

  3. MOLECULAR CLONING AND ANALYSIS OF THE CRYPTOSPORIDIUM PARVUM AMINOPEPTIDASE N GENE. (R828035)

    Science.gov (United States)

    Cryptosporidium parvum proteases have been associated with release of infective sporozoites from oocysts, and their specific inhibition blocks parasite excystation in vitro. Additionally, proteases have been implicated in the processing of parasite adhesion molecules fo...

  4. Interactions between Cryptosporidium parvum and the Intestinal Ecosystem

    KAUST Repository

    Douvropoulou, Olga

    2017-04-01

    Cryptosporidium parvum is an apicomplexan protozoan parasite commonly causing diarrhea, particularly in infants in developing countries. The research challenges faced in the development of therapies against Cryptosporidium slow down the process of drug discovery. However, advancement of knowledge towards the interactions of the intestinal ecosystem and the parasite could provide alternative approaches to tackle the disease. Under this perspective, the primary focus of this work was to study interactions between Cryptosporidium parvum and the intestinal ecosystem in a mouse model. Mice were treated with antibiotics with different activity spectra and the resulted perturbation of the native gut microbiota was identified by microbiome studies. In particular, 16S amplicon sequencing and Whole Genome Sequencing (WGS) were used to determine the bacterial composition and the genetic repertoire of the fecal microbial communities in the mouse gut. Following alteration of the microbial communities of mice by application of antibiotic treatment, Cryptosporidium parasites were propagated in mice with perturbed microbiota and the severity of the infection was quantified. This approach enabled the prediction of the functional capacity of the microbial communities in the mouse gut and led to the identification of bacterial taxa that positively or negatively correlate in abundance with Cryptosporidium proliferation.

  5. Structure-activity relationship study of selective benzimidazole-based inhibitors of Cryptosporidium parvum IMPDH

    OpenAIRE

    Kirubakaran, Sivapriya; Gorla, Suresh Kumar; Sharling, Lisa; Zhang, Minjia; Liu, Xiaoping; Ray, Soumya S.; Iain S. MacPherson; Striepen, Boris; Hedstrom, Lizbeth; Gregory D Cuny

    2012-01-01

    Cryptosporidium parasites are important waterborne pathogens of both humans and animals. The C. parvum and C. hominis genomes indicate that the only route to guanine nucleotides is via inosine 5'-monophosphate dehydrogenase (IMPDH). Thus the inhibition of the parasite IMPDH presents a potential strategy for treating Cryptosporidium infections. A selective benzimidazole-based inhibitor of C. parvum IMPDH (CpIMPDH) was previously identified in a high throughput screen. Here we report a structur...

  6. Effects of Enterococcus faecalis CECT 7121 on Cryptosporidium parvum infection in mice

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    Cryptosporidium is an opportunistic protozoan parasite of humans and animals worldwide, causes diarrheal disease that is typically self-limiting in immunocompetent hosts but often life-threatening to immunocompromised individuals. However, there is a lack of completely efficient therapy available. P...

  7. Crystal structure of Cryptosporidium parvum pyruvate kinase.

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    William J Cook

    Full Text Available Pyruvate kinase plays a critical role in cellular metabolism of glucose by serving as a major regulator of glycolysis. This tetrameric enzyme is allosterically regulated by different effector molecules, mainly phosphosugars. In response to binding of effector molecules and substrates, significant structural changes have been identified in various pyruvate kinase structures. Pyruvate kinase of Cryptosporidium parvum is exceptional among known enzymes of protozoan origin in that it exhibits no allosteric property in the presence of commonly known effector molecules. The crystal structure of pyruvate kinase from C. parvum has been solved by molecular replacement techniques and refined to 2.5 Å resolution. In the active site a glycerol molecule is located near the γ-phosphate site of ATP, and the protein structure displays a partially closed active site. However, unlike other structures where the active site is closed, the α6' helix in C. parvum pyruvate kinase unwinds and assumes an extended conformation. In the crystal structure a sulfate ion is found at a site that is occupied by a phosphate of the effector molecule in many pyruvate kinase structures. A new feature of the C. parvum pyruvate kinase structure is the presence of a disulfide bond cross-linking the two monomers in the asymmetric unit. The disulfide bond is formed between cysteine residue 26 in the short N-helix of one monomer with cysteine residue 312 in a long helix (residues 303-320 of the second monomer at the interface of these monomers. Both cysteine residues are unique to C. parvum, and the disulfide bond remained intact in a reduced environment. However, the significance of this bond, if any, remains unknown at this time.

  8. Cryptosporidium parvum has an active hypusine biosynthesis pathway.

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    Mittal, Nimisha; Morada, Marie; Tripathi, Pankaj; Gowri, V S; Mandal, Swati; Quirch, Alison; Park, Myung Hee; Yarlett, Nigel; Madhubala, Rentala

    2014-06-01

    The protozoan parasite Cryptosporidium parvum causes severe enteric infection and diarrheal disease with substantial morbidity and mortality in untreated AIDS patients and children in developing or resource-limited countries. No fully effective treatment is available. Hypusination of eIF5A is an important post-translational modification essential for cell proliferation. This modification occurs in a two step process catalyzed by deoxyhypusine synthase (DHS) followed by deoxyhypusine hydroxylase. An ORF of 1086bp was identified in the C. parvum (Cp) genome which encodes for a putative polypeptide of 362 amino acids. The recombinant CpDHS protein was purified to homogeneity and used to probe the enzyme's mechanism, structure, and inhibition profile in a series of kinetic experiments. Sequence analysis and structural modeling of CpDHS were performed to probe differences with respect to the DHS of other species. Unlike Leishmania, Trypanosomes and Entamoeba, Cryptosporidium contains only a single gene for DHS. Phylogenetic analysis shows that CpDHS is more closely related to apicomplexan DHS than kinetoplastid DHS. Important residues that are essential for the functioning of the enzyme including NAD(+) binding residues, spermidine binding residues and the active site lysine are conserved between CpDHS and human DHS. N(1)-guanyl-1,7-diaminoheptane (GC7), a potent inhibitor of DHS caused an effective inhibition of infection and growth of C. parvum in HCT-8 cells. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. Development of Cryptosporidium parvum-Induced Gastrointestinal Neoplasia in Severe Combined Immunodeficiency (SCID) Mice: Severity of Lesions Is Correlated with Infection Intensity

    Science.gov (United States)

    Certad, Gabriela; Creusy, Colette; Ngouanesavanh, Tramy; Guyot, Karine; Gantois, Nausicaa; Mouray, Anthony; Chassat, Thierry; Flament, Nicolas; Fleurisse, Laurence; Pinon, Anthony; Delhaes, Laurence; Dei-Cas, Eduardo

    2010-01-01

    We reported previously that Cryptosporidium parvum was able to induce intestinal tumors in severe combined immunodeficiency (SCID) mice treated with corticoids. To further characterize this Cryptosporidium-induced cell transformation, SCID mice treated with dexamethasone were challenged with C. parvum oocysts, and euthanatized sequentially after infection for histologic examination. Ki-67 was used as a marker of cellular proliferation. Our previous results were confirmed, and it was also found that mice receiving higher inocula (106–107) experienced more severe neoplastic development. Additionally, neoplastic changes were observed not only in the caecum but also in the stomach and duodenum of some animals. Interestingly, SCID mice (6/6) inoculated with 105–107 oocysts showed high grade intraepithelial neoplasia or adenomas with high grade dysplasia in the caecum after Day 46 post-infection (PI). Immunohistochemistry for Ki-67 staining indicated the neoplastic process associated to cryptosporidiosis, and evidenced the first immunohistochemical alterations at early stages of the process, even at 3 weeks PI. PMID:20134002

  10. Binding and activation of human and mouse complement by Cryptosporidium parvum (Apicomplexa) and susceptibility of C1q- and MBL-deficient mice to infection.

    Science.gov (United States)

    Petry, Franz; Jakobi, Vera; Wagner, Swen; Tessema, Tesfaye Sisay; Thiel, Steffen; Loos, Michael

    2008-07-01

    Cryptosporidium parvum is a protozoan parasite (Apicomplexa) that causes gastrointestinal disease in animals and humans. Whereas immunocompetent hosts can limit the infection within 1 or 2 weeks, immunocompromised individuals develop a chronic, life-threatening disease. The importance of the adaptive cellular immune response, with CD4+ T-lymphocytes being the major players, has been clearly demonstrated. Several non-adaptive immune mechanisms have been suggested to contribute to the host defence, such as interferon-gamma (IFN-gamma) from NK cells, certain chemokines, beta-defensins and pro-inflammatory cytokines, but the influence of the complement systems has been less well studied. We analysed the in vitro binding and activation of the human and mouse complement systems and tested the susceptibility to infection in complement-deficient mouse strains. We found that C. parvum can activate both the classical and lectin pathways, leading to the deposition of C3b on the parasite. Using real-time PCR, parasite development could be demonstrated in adult mice lacking mannan-binding lectin (MBL-A/C-/-) but not in mice lacking complement factor C1q (C1qA-/-) or in wild type C57BL/6 mice. The contribution of the complement system and the lectin pathway in particular to the host defence against cryptosporidiosis may become apparent in situations of immunodeficiency such as HIV infections or in early childhood.

  11. Structure-activity relationship study of selective benzimidazole-based inhibitors of Cryptosporidium parvum IMPDH

    Science.gov (United States)

    Kirubakaran, Sivapriya; Gorla, Suresh Kumar; Sharling, Lisa; Zhang, Minjia; Liu, Xiaoping; Ray, Soumya S.; MacPherson, Iain S.; Striepen, Boris; Hedstrom, Lizbeth; Cuny, Gregory D.

    2012-01-01

    Cryptosporidium parasites are important waterborne pathogens of both humans and animals. The C. parvum and C. hominis genomes indicate that the only route to guanine nucleotides is via inosine 5'-monophosphate dehydrogenase (IMPDH). Thus the inhibition of the parasite IMPDH presents a potential strategy for treating Cryptosporidium infections. A selective benzimidazole-based inhibitor of C. parvum IMPDH (CpIMPDH) was previously identified in a high throughput screen. Here we report a structure-activity relationship study of benzimidazole-based compounds that resulted in potent and selective inhibitors of CpIMPDH. Several compounds display potent antiparasitic activity in vitro. PMID:22310229

  12. Over-expression and localization of a host protein on the membrane of Cryptosporidium parvum infected epithelial cells.

    Science.gov (United States)

    Yang, Yi-Lin; Serrano, Myrna G; Sheoran, Abhineet S; Manque, Patricio A; Buck, Gregory A; Widmer, Giovanni

    2009-11-01

    The genus Cryptosporidium includes several species of intestinal protozoan parasites which multiply in intestinal epithelial cells. The impact of this infection on the transcriptome of cultured host cells was investigated using DNA microarray hybridizations. The expression of 14 genes found to be consistently up- or down-regulated in infected cell monolayers was validated with RT PCR. Using immunofluorescence we examined the expression of Protease Activated Receptor-2, which is encoded by one of the up-regulated genes. In infected cells this receptor localized to the host cell membrane which covers the intracellular trophozoites and meronts. This observation indicates that the composition of the host cell membrane is affected by the developing trophozoite, a phenomenon which has not been described previously.

  13. Hydrophobic and electrostatic cell surface properties of Cryptosporidium parvum.

    OpenAIRE

    Drozd, C; Schwartzbrod, J

    1996-01-01

    Microbial adhesion to hydrocarbons and microelectrophoresis were investigated in order to characterize the surface properties of Cryptosporidium parvum. Oocysts exhibited low removal rates by octane (only 20% on average), suggesting that the Cryptosporidium sp. does not demonstrate marked hydrophobic properties. A zeta potential close to -25 mV at pH 6 to 6.5 in deionized water was observed for the parasite. Measurements of hydrophobicity and zeta potential were performed as a function of pH ...

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

    Science.gov (United States)

    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

  15. Effects of ozone, chlorine dioxide, chlorine, and monochloramine on Cryptosporidium parvum oocyst viability.

    OpenAIRE

    Korich, D. G.; Mead, J R; Madore, M S; Sinclair, N A; Sterling, C R

    1990-01-01

    Purified Cryptosporidium parvum oocysts were exposed to ozone, chlorine dioxide, chlorine, and monochloramine. Excystation and mouse infectivity were comparatively evaluated to assess oocyst viability. Ozone and chlorine dioxide more effectively inactivated oocysts than chlorine and monochloramine did. Greater than 90% inactivation as measured by infectivity was achieved by treating oocysts with 1 ppm of ozone (1 mg/liter) for 5 min. Exposure to 1.3 ppm of chlorine dioxide yielded 90% inactiv...

  16. Molecular characterization of Danish Cryptosporidium parvum isolates

    DEFF Research Database (Denmark)

    Enemark, Heidi L.; Ahrens, Peter; Juel, Cynthia Dawn

    2002-01-01

    was characterized as C. meleagridis. The porcine Cryptosporidium isolates (N = 4) revealed a pattern which was genetically distinct from human and bovine isolates. Cryptosporidium in a hedgehog (Erinaceus europaeus L.) was identified for the first time. By microsatellite sequencing the hedgehog isolate showed...

  17. Preliminary molecular characterization of Cryptosporidium parvum isolates of wildlife rodents from Poland.

    Science.gov (United States)

    Bajer, A; Cacciò, S; Bednarska, M; Behnke, J M; Pieniazek, N J; Sinski, E

    2003-10-01

    Isolates of Cryptosporidium were collected from 3 species of woodland and field rodents (Clethrionomys glareolus, Microtus arvalis, and Apodemus flavicollis) and were characterized by polymerase chain reaction amplification and sequencing of fragments of the oocyst wall protein (COWP) gene and of the 18S ribosomal RNA gene. Sequence analysis of these markers revealed that the animals were infected with C. parvum, and that the genotype involved was almost identical to the mouse genotype previously described from Mus musculus. Thus, small rodents should be considered as an important reservoir of C. parvum genotypes closely related to the zoonotic genotype 2 and potentially hazardous to humans.

  18. Isolation of Small Number of Cryptosporidium parvum Oocyst Using Immunochromatography.

    Directory of Open Access Journals (Sweden)

    Elahe Ebrahimzade

    2014-12-01

    Full Text Available Cryptosporidium parvum causes severe gastroenteritis in immunocompromised human and new borne animals. The organism can be transmitted through water. Since small number of C. parvum is infectious, the aim of the present study was to develop a chromatography method for the isolation of C. parvum oocyst in samples with limited number of oocysts.Antibody was prepared against whole antigen from C. parvum oocysts, the achieved Ab bound to the sepharose 4B and used for the isolation of oocysts. Antibody against P23 bound to the sepharose 4B, used also for the isolation of C. parvum oocyst. In comparison to these both methods, 2 traditional methods (Salt floatation and 55% sucrose floatation were also performed.Both chromatography methods could bind oocysts with capacity depends on the column size. The isolated oocysts were free of bacteria. Our results showed that the traditional methods are useful for the isolation of oocysts from feces, in its smear stained with ziehl-nelsen, at least 3 oocyts are detectable in each microscopic field under 1000 X magnification. In contrast to the chromatography methods, the bacterial contamination was always observed in oocysts isolated with traditional methods.Immunochromatography could be used for the successful isolation of C. parvum oocysts from the samples containing limited number of oocysts.

  19. [Cryptosporidium parvum in wild gastropods as bioindicators of fecal contamination in terrestrial ecosystems].

    Science.gov (United States)

    Neira O, Patricia; Muñoz S, Nelson; Stanley V, Bárbara; Gosh C, Marianne; Rosales L, M José

    2010-06-01

    Cryptosporidium sp oocysts were detected in snails (Helix aspersa Miller) and slug (Deroceras reticulatum Miller) from the Valparaiso Region, Chile. Snails and slug were collected from public squares and private domestic gardens. Cryptosporidium sp oocysts were recovered from faeces of both species. Ziehl Neelsen stain, nested PCR, and sequencing analysis demonstrated a profile similar to that described for genotype C or 2 of the parasite. These results demonstrate that snails and slug could act as a reservoir and mechanic vector of C. parvum infection for humans and animals. Moreover, gastropods could serve as bioindicators of fecal soil contamination.

  20. Effect of High-Rate Algal Ponds on Viability of Cryptosporidium parvum Oocysts

    Science.gov (United States)

    Araki, S.; Martín-Gomez, S.; Bécares, E.; De Luis-Calabuig, E.; Rojo-Vazquez, F.

    2001-01-01

    The physicochemical conditions of high-rate algal ponds were responsible for a more than 97% reduction in the infectivity of Cryptosporidium parvum oocysts in neonatal mice. The use of semipermeable bags of cellulose showed that pH, ammonia, and/or light seems to be a major factor for the inactivation of oocysts in wastewater, supporting the importance of alga-based systems for safer reuse of treated wastewater. PMID:11425762

  1. Cryptosporidium parvum-induced ileo-caecal adenocarcinoma and Wnt signaling in a mouse model

    Directory of Open Access Journals (Sweden)

    Sadia Benamrouz

    2014-06-01

    Full Text Available Cryptosporidium species are apicomplexan protozoans that are found worldwide. These parasites constitute a large risk to human and animal health. They cause self-limited diarrhea in immunocompetent hosts and a life-threatening disease in immunocompromised hosts. Interestingly, Cryptosporidium parvum has been related to digestive carcinogenesis in humans. Consistent with a potential tumorigenic role of this parasite, in an original reproducible animal model of chronic cryptosporidiosis based on dexamethasone-treated or untreated adult SCID mice, we formerly reported that C. parvum (strains of animal and human origin is able to induce digestive adenocarcinoma even in infections induced with very low inoculum. The aim of this study was to further characterize this animal model and to explore metabolic pathways potentially involved in the development of C. parvum-induced ileo-caecal oncogenesis. We searched for alterations in genes or proteins commonly involved in cell cycle, differentiation or cell migration, such as β-catenin, Apc, E-cadherin, Kras and p53. After infection of animals with C. parvum we demonstrated immunohistochemical abnormal localization of Wnt signaling pathway components and p53. Mutations in the selected loci of studied genes were not found after high-throughput sequencing. Furthermore, alterations in the ultrastructure of adherens junctions of the ileo-caecal neoplastic epithelia of C. parvum-infected mice were recorded using transmission electron microscopy. In conclusion, we found for the first time that the Wnt signaling pathway, and particularly the cytoskeleton network, seems to be pivotal for the development of the C. parvum-induced neoplastic process and cell migration of transformed cells. Furthermore, this model is a valuable tool in understanding the host-pathogen interactions associated with the intricate infection process of this parasite, which is able to modulate host cytoskeleton activities and several host

  2. Effects of ozone, chlorine dioxide, chlorine, and monochloramine on Cryptosporidium parvum oocyst viability

    Energy Technology Data Exchange (ETDEWEB)

    Korich, D.G.; Mead, J.R.; Madore, M.S.; Sinclair, N.A.; Sterling, C.R. (Univ. of Arizona, Tucson (USA))

    1990-05-01

    Purified Cryptosporidium parvum oocysts were exposed to ozone, chlorine dioxide, chlorine, and monochloramine. Excystation and mouse infectivity were comparatively evaluated to assess oocyst viability. Ozone and chlorine dioxide more effectively inactivated oocysts than chlorine and monochloramine did. Greater than 90% inactivation as measured by infectivity was achieved by treating oocysts with 1 ppm of ozone (1 mg/liter) for 5 min. Exposure to 1.3 ppm of chlorine dioxide yielded 90% inactivation after 1 h, while 80 ppm of chlorine and 80 ppm of monochloramine required approximately 90 min for 90% inactivation. The data indicate that C. parvum oocysts are 30 times more resistant to ozone and 14 times more resistant to chlorine dioxide than Giardia cysts exposed to these disinfectants under the same conditions. With the possible exception of ozone, the use of disinfectants alone should not be expected to inactivate C. parvum oocysts in drinking water.

  3. Effects of ozone, chlorine dioxide, chlorine, and monochloramine on Cryptosporidium parvum oocyst viability.

    Science.gov (United States)

    Korich, D G; Mead, J R; Madore, M S; Sinclair, N A; Sterling, C R

    1990-01-01

    Purified Cryptosporidium parvum oocysts were exposed to ozone, chlorine dioxide, chlorine, and monochloramine. Excystation and mouse infectivity were comparatively evaluated to assess oocyst viability. Ozone and chlorine dioxide more effectively inactivated oocysts than chlorine and monochloramine did. Greater than 90% inactivation as measured by infectivity was achieved by treating oocysts with 1 ppm of ozone (1 mg/liter) for 5 min. Exposure to 1.3 ppm of chlorine dioxide yielded 90% inactivation after 1 h, while 80 ppm of chlorine and 80 ppm of monochloramine required approximately 90 min for 90% inactivation. The data indicate that C. parvum oocysts are 30 times more resistant to ozone and 14 times more resistant to chlorine dioxide than Giardia cysts exposed to these disinfectants under the same conditions. With the possible exception of ozone, the use of disinfectants alone should not be expected to inactivate C. parvum oocysts in drinking water. PMID:2339894

  4. Serological detection and epidemiology of Neospora caninum and Cryptosporidium parvum antibodies in cattle in southern Egypt.

    Science.gov (United States)

    Fereig, Ragab M; AbouLaila, Mahmoud Rezk; Mohamed, Samy G A; Mahmoud, Hassan Y A H; Ali, Alsagher O; Ali, Asmaa F; Hilali, Mosaad; Zaid, Anis; Mohamed, Adel Elsayed Ahmed; Nishikawa, Yoshifumi

    2016-10-01

    Neospora caninum and Cryptosporidium parvum are intracellular protozoan parasites that are distributed worldwide and of major economical concern in cattle industry. N. caninum can cause abortion storms and high culling rates, whereas C. parvum has zoonotic implications and can cause diarrhea in calves. There are currently no data on the prevalence of neosporosis and cryptosporidiosis in humans or animals in southern Egypt. Prevalence of these two infections was determined in a sample of cattle from two different areas in southern Egypt, Sohag and Qena, using enzyme-linked immunosorbent assay. A total 301 cattle were sampled, of which 18.9% were positive for N. caninum, 35.9% were positive for C. parvum and 10.0% were positive for both. Geographical location and breeding system were considered as potential risk factors for C. parvum infection. A higher prevalence of infection was identified on small scale farms, compared with larger, intensive systems, with a prevalence of 50.2% compared with 37.8%, respectively. Animals in Sohag had a significantly higher prevalence compared with Qena, with a seroprevalence of 46.1% compared with 31.6%, respectively. In brief, marked seroprevalence recorded in this study indicates a high incidence of N. caninum and C. parvum infections in cattle, and this necessitates the application of more effective strategies for combating these types of infections on farms in Egypt.

  5. Chlorine Dioxide Inactivation of Cryptosporidium parvum Oocysts and Bacterial Spore Indicators

    Science.gov (United States)

    Chauret, Christian P.; Radziminski, Chris Z.; Lepuil, Michael; Creason, Robin; Andrews, Robert C.

    2001-01-01

    Cryptosporidium parvum, which is resistant to chlorine concentrations typically used in water treatment, is recognized as a significant waterborne pathogen. Recent studies have demonstrated that chlorine dioxide is a more efficient disinfectant than free chlorine against Cryptosporidium oocysts. It is not known, however, if oocysts from different suppliers are equally sensitive to chlorine dioxide. This study used both a most-probable-number–cell culture infectivity assay and in vitro excystation to evaluate chlorine dioxide inactivation kinetics in laboratory water at pH 8 and 21°C. The two viability methods produced significantly different results (P < 0.05). Products of disinfectant concentration and contact time (Ct values) of 1,000 mg · min/liter were needed to inactivate approximately 0.5 log10 and 2.0 log10 units (99% inactivation) of C. parvum as measured by in vitro excystation and cell infectivity, respectively, suggesting that excystation is not an adequate viability assay. Purified oocysts originating from three different suppliers were evaluated and showed marked differences with respect to their resistance to inactivation when using chlorine dioxide. Ct values of 75, 550, and 1,000 mg · min/liter were required to achieve approximately 2.0 log10 units of inactivation with oocysts from different sources. Finally, the study compared the relationship between easily measured indicators, including Bacillus subtilis (aerobic) spores and Clostridium sporogenes (anaerobic) spores, and C. parvum oocysts. The bacterial spores were found to be more sensitive to chlorine dioxide than C. parvum oocysts and therefore could not be used as direct indicators of C. parvum inactivation for this disinfectant. In conclusion, it is suggested that future studies address issues such as oocyst purification protocols and the genetic diversity of C. parvum, since these factors might affect oocyst disinfection sensitivity. PMID:11425712

  6. Chlorine dioxide inactivation of Cryptosporidium parvum oocysts and bacterial spore indicators.

    Science.gov (United States)

    Chauret, C P; Radziminski, C Z; Lepuil, M; Creason, R; Andrews, R C

    2001-07-01

    Cryptosporidium parvum, which is resistant to chlorine concentrations typically used in water treatment, is recognized as a significant waterborne pathogen. Recent studies have demonstrated that chlorine dioxide is a more efficient disinfectant than free chlorine against Cryptosporidium oocysts. It is not known, however, if oocysts from different suppliers are equally sensitive to chlorine dioxide. This study used both a most-probable-number-cell culture infectivity assay and in vitro excystation to evaluate chlorine dioxide inactivation kinetics in laboratory water at pH 8 and 21 degrees C. The two viability methods produced significantly different results (P < 0.05). Products of disinfectant concentration and contact time (Ct values) of 1,000 mg. min/liter were needed to inactivate approximately 0.5 log(10) and 2.0 log(10) units (99% inactivation) of C. parvum as measured by in vitro excystation and cell infectivity, respectively, suggesting that excystation is not an adequate viability assay. Purified oocysts originating from three different suppliers were evaluated and showed marked differences with respect to their resistance to inactivation when using chlorine dioxide. Ct values of 75, 550, and 1,000 mg. min/liter were required to achieve approximately 2.0 log(10) units of inactivation with oocysts from different sources. Finally, the study compared the relationship between easily measured indicators, including Bacillus subtilis (aerobic) spores and Clostridium sporogenes (anaerobic) spores, and C. parvum oocysts. The bacterial spores were found to be more sensitive to chlorine dioxide than C. parvum oocysts and therefore could not be used as direct indicators of C. parvum inactivation for this disinfectant. In conclusion, it is suggested that future studies address issues such as oocyst purification protocols and the genetic diversity of C. parvum, since these factors might affect oocyst disinfection sensitivity.

  7. COMPARISON OF TISSUE CULTURE AND ANIMAL MODELS FOR ASSESSMENT OF CRYPTOSPRIDIUM PARVUM INFECTION

    Science.gov (United States)

    Data from three different disinfection studies using both cell culture and mouse infectivity to assess Cryptosporidium parvum inactivation were evaluated in a total of 35 comparison including process controls and treated samples. C. parvum infectivity in the in vitro FDM-MPN assa...

  8. Comparative efficacy of conventional primer sets in detection of Cryptosporidium parvum for diagnostic use.

    Science.gov (United States)

    Kar, Sirri; Daugschies, Arwid; Bangoura, Berit

    2010-02-01

    In this study, the sensitivity and specificity of different previously described primer sets for Cryptosporidium parvum detection by polymerase chain reaction (PCR) was evaluated. For this purpose, the primer sets defined by Cacciò et al. (FEMS Microbiol Lett 170(1):173-179, 1999) (tub), Widmer et al. (Appl Environ Microbiol 64(11):4477-4481, 1998) (btub) and Rochelle et al. (Appl Environ Microbiol 63:2029-2037, 1997) (cphsp), respectively, were used. Deoxyribonucleic acid (DNA) was isolated from three different sample materials: (1) from the faeces of an experimentally C. parvum-infected calf, (2) from purified C. parvum oocysts, and (3) from C. parvum-infected HCT-8 cell cultures. The DNA samples were subjected to PCR reactions with each of the three given primer sets to investigate sensitivity and suitability for routine use. The primers described by Cacciò et al. (FEMS Microbiol Lett 170(1):173-179, 1999) (TUB) were superior regarding sensitivity and specificity in terms of detection of C. parvum in faeces, in purified oocysts and also in cell culture, and may thus be applied for routine diagnostic use in common sample materials.

  9. Putative cis-regulatory elements associated with heat shock genes activated during excystation of Cryptosporidium parvum.

    Directory of Open Access Journals (Sweden)

    Benjamin Cohn

    Full Text Available BACKGROUND: Cryptosporidiosis is a ubiquitous infectious disease, caused by the protozoan parasites Cryptosporidium hominis and C. parvum, leading to acute, persistent and chronic diarrhea worldwide. Although the complications of this disease can be serious, even fatal, in immunocompromised patients of any age, they have also been found to lead to long term effects, including growth inhibition and impaired cognitive development, in infected immunocompetent children. The Cryptosporidium life cycle alternates between a dormant stage, the oocyst, and a highly replicative phase that includes both asexual vegetative stages as well as sexual stages, implying fine genetic regulatory mechanisms. The parasite is extremely difficult to study because it cannot be cultured in vitro and animal models are equally challenging. The recent publication of the genome sequence of C. hominis and C. parvum has, however, significantly advanced our understanding of the biology and pathogenesis of this parasite. METHODOLOGY/PRINCIPAL FINDINGS: Herein, our goal was to identify cis-regulatory elements associated with heat shock response in Cryptosporidium using a combination of in silico and real time RT-PCR strategies. Analysis with Gibbs-Sampling algorithms of upstream non-translated regions of twelve genes annotated as heat shock proteins in the Cryptosporidium genome identified a highly conserved over-represented sequence motif in eleven of them. RT-PCR analyses, described herein and also by others, show that these eleven genes bearing the putative element are induced concurrent with excystation of parasite oocysts via heat shock. CONCLUSIONS/SIGNIFICANCE: Our analyses suggest that occurrences of a motif identified in the upstream regions of the Cryptosporidium heat shock genes represent parts of the transcriptional apparatus and function as stress response elements that activate expression of these genes during excystation, and possibly at other stages in the life

  10. Excystation of Cryptosporidium parvum at temperatures that are reached during solar water disinfection.

    Science.gov (United States)

    Gómez-Couso, H; Fontán-Sainz, M; Fernández-Alonso, J; Ares-Mazás, E

    2009-04-01

    Species belonging to the genera Cryptosporidium are recognized as waterborne pathogens. Solar water disinfection (SODIS) is a simple method that involves the use of solar radiation to destroy pathogenic microorganisms that cause waterborne diseases. A notable increase in water temperature and the existence of a large number of empty or partially excysted (i.e. unviable) oocysts have been observed in previous SODIS studies with water experimentally contaminated with Cryptosporidium parvum oocysts under field conditions. The aim of the present study was to evaluate the effect of the temperatures that can be reached during exposure of water samples to natural sunlight (37-50 degrees C), on the excystation of C. parvum in the absence of other stimuli. In samples exposed to 40-48 degrees C, a gradual increase in the percentage of excystation was observed as the time of exposure increased and a maximum of 53.81% of excystation was obtained on exposure of the water to a temperature of 46 degrees C for 12 h (versus 8.80% initial isolate). Under such conditions, the oocyst infectivity evaluated in a neonatal murine model decreased statistically with respect to the initial isolate (19.38% versus 100%). The results demonstrate the important effect of the temperature on the excystation of C. parvum and therefore on its viability and infectivity.

  11. Vaccination with pcDNA3-15/60 Naked DNA Encoding the Surface Proteinof Sporozoites in Cryptosporidium parvum

    Institute of Scientific and Technical Information of China (English)

    HEHong-xuan; ZHANGXi-chen; YINJi-gang; LIJian-hua; YANGJu

    2004-01-01

    The CP15/60 gene encoding the CP15/60 surface protein of sporozoites in Cryptosporidium parvum was obtained by PCR so as to research the nucleic vaccine against C.parvum. The eukaryotic expressing vector pcDNA3-15/60 was constructed by inserting CP15/60 gene into pcDNA3 (+) in Xho Ⅰ and EcoR Ⅰ. A vaccination protocol was the adult pregnant goats inoculated intranasally with the pcDNA3-15/60 plasmid and their offspring were infected with C.parvum oocysts. The results showed that the pcDNA3-15/60 plasmid can induce the immune response of goats and the vaccinated goats can transfer the immunity to offspring conferring protection against C.parvum infection. These suggested that the recombinant plasmid could be a DNA vaccine candidate.

  12. Vaccination with pcDNA3-15/60 Naked DNA Encoding the Surface Protein of Sporozoites in Cryptosporidium parvum

    Institute of Scientific and Technical Information of China (English)

    HE Hong-xuan; ZHANG Xi-chen; YIN Ji-gang; LI Jian-hua; YANG Ju

    2004-01-01

    The CP15/60 gene encoding the CP15/60 surface protein of sporozoites in Cryptosporidium parvum was obtained by PCR so as to research the nucleic vaccine against C.parvum. The eukaryotic expressing vector pcDNA3-15/60 was constructed by inserting CP15/60 gene into pcDNA3 (+) in Xho Ⅰ and EcoR Ⅰ. A vaccination protocol was the adult pregnant goats inoculated intranasally with the pcDNA3-15/60 plasmid and their offspring were infected with C.parvum oocysts. The results showed that the pcDNA3-15/60 plasmid can induce the immune response of goats and the vaccinated goats can transfer the immunity to offspring conferring protection against C.parvum infection. These suggested that the recombinant plasmid could be a DNA vaccine candidate.

  13. Viability of Cryptosporidium parvum during ensilage of perennial ryegrass.

    Science.gov (United States)

    Merry, R J; Mawdsley, J L; Brooks, A E; Davies, D R

    1997-01-01

    The survival of Cryptosporidium parvum during ensilage of perennial ryegrass was examined in laboratory silos with herbage prepared in one of three different ways; either untreated, inoculated with a strain of Lactobacillus plantarum or by direct acidification with formic acid. The pH values of all silages initially fell below 4.5, but only formic acid-treated silage remained stable at less than pH 4 after 106 d, with the pH of the untreated and inoculant-treated silages rising to above 6. The formic acid-treated silage had a high lactic acid concentration (109 g kg-1 dry matter (DM)) and low concentrations of propionic and butyric acids after 106 d. However, the untreated and inoculant-treated silages showed an inverse relationship, with low lactic acid concentrations and high concentrations of acetic, propionic and butyric acids. These silages also contained ammonia-N concentrations in excess of 9 g kg-1 DM. In terms of the viability of Cryptosporidium parvum oocysts very few differences were seen after 14 d of ensilage with ca 50% remaining viable, irrespective of treatment and total numbers had declined from the initial level of 5.9 x 10(4) to 1 x 10(4) g(-1) fresh matter. Total oocyst numbers remained approximately the same until the end of the ensiling period, with the percentage of viable oocysts declining to 46, 41 and 32% respectively for formic acid, inoculant and untreated silages. The results are discussed in terms of changes occurring during the silage fermentation, in particular the products which may influence the survival of Cryptosporidium and implications for agricultural practice and the health of silage fed livestock.

  14. Detection of Giardia lamblia and Cryptosporidium parvum by direct immunofluorescence assay in stool specimen.

    Science.gov (United States)

    Rahman, M M; Hossain, M A; Paul, S K; Ahmed, S; Islam, A; Ehsan, M A; Alam, M M; Kabir, M R; Sarkar, S R

    2014-07-01

    Giardia and Cryptosporidium are the pathogens which transmitted through contaminated soil and contaminated water are significant causes of diarrhea and nutritional disorders in institutional and community peoples. Children and immune compromise persons are more vulnerable for these infections. Both Giardiasis and Cryptosporidiosis were included in 2004 as WHO Neglected Disease. So this is a major public health problem in developing countries. The present study was carried out to detect the Giardia and Cryptosporidium from diarrheic or patient having loose stool by Direct Immunofluorescence assay. The study was conducted during July 20012 to February 2013 and the work was done in Mymensingh Medical College in the department of Microbiology and in Bangladesh Agricultural University in the department of Veterinary Medicine. A total of 100 loose stools were collected from school children of different area and hospital under sadar upazilla, Mymensingh. The detection of Giardia lamblia and Cryptosporidium parvum showed the individual prevalence 8% and 4% respectively. The highest cyst/oocyst count was 85,000 and 1,000/gm of stool and the lowest being 100 and 50/gm of stool for Giardiasis and Cryptosporidiosis respectively. The detection rate of Giardia and Cryptosporidium by Immunofluorescence assay was relatively higher than the previous study done in Bangladesh and this was the first report from Bangladesh over human stool specimen using Immunofluorescence assay. So, Immunofluorescence assay could be adapted for rapid and accurate detection of Giardia and Cryptosporidium.

  15. Hydrophobic and electrostatic cell surface properties of Cryptosporidium parvum.

    Science.gov (United States)

    Drozd, C; Schwartzbrod, J

    1996-04-01

    Microbial adhesion to hydrocarbons and microelectrophoresis were investigated in order to characterize the surface properties of Cryptosporidium parvum. Oocysts exhibited low removal rates by octane (only 20% on average), suggesting that the Cryptosporidium sp. does not demonstrate marked hydrophobic properties. A zeta potential close to -25 mV at pH 6 to 6.5 in deionized water was observed for the parasite. Measurements of hydrophobicity and zeta potential were performed as a function of pH and ionic strength or conductivity. Hydrophobicity maxima were observed at extreme pH values, with 40% of adhesion of oocysts to octane. It also appeared that ionic strength (estimated by conductivity) could influence the hydrophobic properties of oocysts. Cryptosporidium oocysts showed a pH-dependent surface charge, with zeta potentials becoming less negative as pH was reduced, starting at -35 mV for alkaline pH and reaching 0 at isoelectric points for pH 2.5. On the other hand, variation of surface charge with respect to conductivity of the suspension tested in this work was quite small. The knowledge of hydrophobic properties and surface charge of the parasite provides information useful in, for example, the choice of various flocculation treatments, membrane filters, and cleaning agents in connection with oocyst recovery.

  16. Immunogold labeling of stages of Cryptosporidium parvum recognized by immunoglobulins in hyperimmune bovine colostrum.

    Science.gov (United States)

    Fayer, R; Barta, J R; Guidry, A J; Blagburn, B L

    1991-06-01

    Ultrathin sections of mouse ileum infected with Cryptosporidium parvum were stained by immunogold techniques. Sections first were stained with polyvalent antibodies in whey from hyperimmune bovine colostrum (HBC), then stained by secondary antibodies in rabbit antibovine IgA, IgM, IgG1, and IgG2, and lastly labeled by goat anti-rabbit gold conjugate. Examination of the immunostained specimens by electron microscopy revealed that each bovine immunoglobulin isotype in the whey recognized antigens in meronts, merozoites, microgametocytes, microgametes, and macrogamonts. Based on these findings it is hypothesized that antigens in all stages of C. parvum provide targets of opportunity for the antiparasitic activity of HBC whey antibodies thereby accounting for its efficacy as an immunotherapeutic agent.

  17. AGING OF CRYPTOSPORIDIUM PARVUM OOCYSTS STUDIED BY MALDI-TOF MS

    Science.gov (United States)

    Cryptosporidium parvum is a protozoan parasite, and it causes a potentially fatal gastrointestinal illness. This water borne pathogen has been the subject of several high profile disease outbreaks in the US and abroad. C. parvum presents challenges for both compliance monitoring ...

  18. Photocatalytic inactivation of Cryptosporidium parvum on nanostructured titanium dioxide films.

    Science.gov (United States)

    Sunnotel, O; Verdoold, R; Dunlop, P S M; Snelling, W J; Lowery, C J; Dooley, J S G; Moore, J E; Byrne, J A

    2010-03-01

    Control of waterborne gastrointestinal parasites represents a major concern to water industries worldwide. In developed countries, pathogens in drinking water supplies are normally removed by sand filtration followed by chemical disinfection. Cryptosporidium spp. are generally resistant to common disinfection techniques and alternative control strategies are being sought. In the current study, the photocatalytic inactivation of C. parvum oocysts was shown to occur in buffer solution (78.4% after 180 min) and surface water (73.7% after 180 min). Viability was assessed by dye exclusion, excystation, direct examination of oocysts and a novel gene expression assay based on lactate dehydrogenase 1 (LDH1) expression levels. Collectively, this confirmed the inactivation of oocysts and scanning electron microscopy (SEM) confirmed cleavage at the suture line of oocyst cell walls, revealing large numbers of empty (ghost) cells after exposure to photocatalytic treatment.

  19. More productive in vitro culture of Cryptosporidium parvum for better study of the intra- and extracellular phases

    Directory of Open Access Journals (Sweden)

    G Perez Cordón

    2007-08-01

    Full Text Available The great difficulties in treating people and animals suffering from cryptosporidiosis have prompted the development of in vitro experimental models. Due to the models of in vitro culture, new extracellular stages of Cryptosporidium have been demonstrated. The development of these extracellular phases depends on the technique of in vitro culture and on the species and genotype of Cryptosporidium used. Here, we undertake the molecular characterization by polymerase chain reaction-restriction fragment lenght polymorphism of different Cryptosporidium isolates from calves, concluding that all are C. parvum of cattle genotype, although differing in the nucleotide at positions 472 and 498. Using these parasites, modified the in vitro culture technique for HCT-8 cells achieving greater multiplication of parasites. The HCT-8 cell cultures, for which the culture had not been renewed in seven days, were infected with C. parvum sporozoites in RPMI-1640 medium with 10% IFBS, CaCl2 and MgCl2 1 mM at pH 7.2. Percentages of cell parasitism were increased with respect to control cultures (71% at 48 h vs 14.5%, even after two weeks (47% vs 1.9%. Also, the percentage of extracellular stages augmented (25.3% vs 1.1% at 96 h. This new model of in vitro culture of C. parvum will enable easier study of the developmental phases of C. parvum in performing new chemotherapeutic assays.

  20. Pomegranate (Punica granatum) peel is effective in a murine model of experimental Cryptosporidium parvum.

    Science.gov (United States)

    Al-Mathal, Ebtisam M; Alsalem, Afaf M

    2012-07-01

    Cryptosporidiosis, a major health issue for neonatal calves, is caused by the parasite Cryptosporidium parvum, which is highly resistant to drug treatments. To date, many anti-parasitic drugs have been tested, but only a few have been shown to be partially effective in treating cryptosporidiosis. Previous studies have indicated that pomegranate (Punica granatum) possesses anti-plasmodium, anti-cestode, and anti-nematode activities. Therefore, the aim of this study was to evaluate the effect of P. granatum peel on suckling mice infected with experimental C. parvum. At 4days of age, 72 neonatal albino mice were randomly divided into five groups: G1: healthy controls, G2: infected/untreated controls, G3: uninfected/distilled water-treated, G4: uninfected/P. granatum peel-treated, and G5: infected/P. granatum peel-treated. Mice were experimentally-infected by oral administration of 1×10(3)C. parvum oocysts per animal. On day 7 post-inoculation (pi), treated mice received an aqueous suspension of P. granatum peel orally (3g/kg body weight). The presence of diarrhea, oocyst shedding, and weight gain/loss, and the histopathology of ileal sections were examined. Infected mice treated with the P. granatum peel suspension showed improvement in all parameters examined. Additionally, these mice did not exhibit any clinical symptoms and no deaths occurred. Oocyst shedding was very significantly reduced in the P. granatum-treated mice by day 14 pi (Parchitecture of villi from the P. granatum-treated mice on day 14 pi showed visible improvement in comparison with the infected/untreated controls, including renewed brush borders, reduced numbers of C. parvum trophozoites, and reduced lymphatic infiltration. On day 28 pi, tissues of the P. granatum-treated mice were very similar to those of healthy control mice. These results suggest that P. granatum peel is a promising anti-coccidial therapeutic treatment that lacks negative side effects.

  1. Two geographically separated food-borne outbreaks in Sweden linked by an unusual Cryptosporidium parvum subtype, October 2010.

    Science.gov (United States)

    Gherasim, A; Lebbad, M; Insulander, M; Decraene, V; Kling, A; Hjertqvist, M; Wallensten, A

    2012-11-15

    The number of sporadic cases of Cryptosporidium identified in the Stockholm county area increased above the expected limit during October 2010. Additionally, two food-borne outbreaks of cryptosporidiosis occurred in two other Swedish cities: Umeå (4 October) and Örebro (9 October). The outbreak investigations did not reveal any responsible food item, however fresh herbs were suspected. Thirty stool samples, originating from all three events, tested positive for Cryptosporidium oocysts. Polymerase chain reaction (PCR) and subsequent restriction fragment length polymorphism (RFLP) revealed that 27 individuals were infected with C. parvum, two with C. hominis, and one with C. felis. Using sequence analysis of the GP60 glycoprotein gene, a polymorphic marker with high intra-species diversity, we identified the same C. parvum subtype IIdA24G1 in samples from both the Umeå outbreak and the Stockholm area cases, thus indicating a possible outbreak in the Stockholm area and establishing a link between these two events. C. parvum IIdA24G1 has not previously been described in connection with a food-borne outbreak. For the outbreak in Örebro, another subtype was identified: C. parvum IIdA20G1e. These findings demonstrate that subtyping C. parvum isolates using GP60 gene amplification can be used to link cases in an outbreak investigation and we recommend its use in future similar events.

  2. Use of a Sentinel System for Field Measurements of Cryptosporidium parvum Oocyst Inactivation in Soil and Animal Waste

    OpenAIRE

    Jenkins, M. B.; Walker, M. J.; Bowman, D. D.; Anthony, L. C.; Ghiorse, W C

    1999-01-01

    A small-volume sentinel chamber was developed to assess the effects of environmental stresses on survival of sucrose-Percoll-purified Cryptosporidium parvum oocysts in soil and animal wastes. Chambers were tested for their ability to equilibrate with external chemical and moisture conditions. Sentinel oocysts were then exposed to stresses of the external environment that affected their viability (potential infectivity), as indicated by results of a dye permeability assay. Preliminary laborato...

  3. Detecting Cryptosporidium parvum and Giardia lamblia by coagulation concentration and real-time PCR quantification

    Institute of Scientific and Technical Information of China (English)

    Huining ZHANG; Xiaohu Zhang; Shuting ZHANG; Bo WEI; Qipei JIANG; Xin YU

    2013-01-01

    Rapid and sensitive pathogen detection methods are essential for the effective functioning of the water treatment industry, yet for many pathogens, effective detection and removal methods are still lacking. Cryptosporidium parvum oocysts and Giardia lamblia cysts are two of the most common waterborne pathogens currently infecting the water supply. In this study, a new method was developed for the detection of Cryptosporidium parvum oocysts and Giardia lamblia cysts. The method includes multi-steps as coagulation concentration of (oo)cysts in water, the dissolution of the resulting flocs into a small volume using acid, filtration of the (oo)cysts solution, and DNA extraction, purification, and examination using real-time PCR. The method was tested using spiked tap water and reservoir water as references, and the mean recovery ranged from 19.6% to 97.6% for oocysts, and from 51.4% to 98.7% for cysts. The method is economical and convenient, and is especially suitable for relatively high turbidity surface water.

  4. A unique hexokinase in Cryptosporidium parvum, an apicomplexan pathogen lacking the Krebs cycle and oxidative phosphorylation.

    Science.gov (United States)

    Yu, Yonglan; Zhang, Haili; Guo, Fengguang; Sun, Mingfei; Zhu, Guan

    2014-09-01

    Cryptosporidium parvum may cause virtually untreatable infections in AIDS patients, and is recently identified as one of the top four diarrheal pathogens in children in developing countries. Cryptosporidium differs from other apicomplexans (e.g., Plasmodium and Toxoplasma) by lacking many metabolic pathways including the Krebs cycle and cytochrome-based respiratory chain, thus relying mainly on glycolysis for ATP production. Here we report the molecular and biochemical characterizations of a hexokinase in C. parvum (CpHK). Our phylogenetic reconstructions indicated that apicomplexan hexokinases including CpHK were highly divergent from those of humans and animals (i.e., at the base of the eukaryotic clade). CpHK displays unique kinetic features that differ from those in mammals and Toxoplasma gondii (TgHK) in the preference towards various hexoses and its capacity to use ATP and other NTPs. CpHK also displays substrate inhibition by ATP. Moreover, 2-deoxy-D-glucose (2DG) could not only inhibit the CpHK activity, but also the parasite growth in vitro at concentrations nontoxic to host cells (IC(50) = 0.54 mM). While the exact action of 2-deoxy-D-glucose on the parasite is subject to further verification, our data suggest that CpHK and the glycolytic pathway may be explored for developing anti-cryptosporidial therapeutics.

  5. In vitro inhibitory effects of plant-derived by-products against Cryptosporidium parvum

    Directory of Open Access Journals (Sweden)

    Teichmann Klaus

    2016-01-01

    Full Text Available Disposal of organic plant wastes and by-products from the food or pharmaceutical industries usually involves high costs. In the present study, 42 samples derived from such by-products were screened in vitro against Cryptosporidium parvum, a protozoan parasite that may contaminate drinking water and cause diarrhoea. The novel bioassay was previously established in the microtitre plate format. Human ileocaecal adenocarcinoma (HCT-8 cell cultures were seeded with C. parvum oocysts and parasite development was monitored by an indirect fluorescent antibody technique (IFAT and microscopic assessment for clusters of secondary infection (CSI. Minimum inhibitory concentrations (MICs and potential detrimental effects on the host cells were determined. An ethanolic extract from olive (Olea europaea pomace, after oil pressing and phenol recovery, reproducibly inhibited C. parvum development (MIC = 250–500 μg mL−1, IC50 = 361 (279–438 μg mL−1, IC90 = 467 (398–615 μg mL−1. Accordingly, tyrosol, hydroxytyrosol, trans-coniferyl alcohol and oleuropein were selected as reference test compounds, but their contributions to the observed activity of the olive pomace extract were insignificant. The established test system proved to be a fast and efficient assay for identifying anti-cryptosporidial activities in biological waste material and comparison with selected reference compounds.

  6. In vitro inhibitory effects of plant-derived by-products against Cryptosporidium parvum

    Science.gov (United States)

    Teichmann, Klaus; Kuliberda, Maxime; Schatzmayr, Gerd; Pacher, Thomas; Zitterl-Eglseer, Karin; Joachim, Anja; Hadacek, Franz

    2016-01-01

    Disposal of organic plant wastes and by-products from the food or pharmaceutical industries usually involves high costs. In the present study, 42 samples derived from such by-products were screened in vitro against Cryptosporidium parvum, a protozoan parasite that may contaminate drinking water and cause diarrhoea. The novel bioassay was previously established in the microtitre plate format. Human ileocaecal adenocarcinoma (HCT-8) cell cultures were seeded with C. parvum oocysts and parasite development was monitored by an indirect fluorescent antibody technique (IFAT) and microscopic assessment for clusters of secondary infection (CSI). Minimum inhibitory concentrations (MICs) and potential detrimental effects on the host cells were determined. An ethanolic extract from olive (Olea europaea) pomace, after oil pressing and phenol recovery, reproducibly inhibited C. parvum development (MIC = 250–500 μg mL−1, IC50 = 361 (279–438) μg mL−1, IC90 = 467 (398–615) μg mL−1). Accordingly, tyrosol, hydroxytyrosol, trans-coniferyl alcohol and oleuropein were selected as reference test compounds, but their contributions to the observed activity of the olive pomace extract were insignificant. The established test system proved to be a fast and efficient assay for identifying anti-cryptosporidial activities in biological waste material and comparison with selected reference compounds. PMID:27627637

  7. Parasites and malignancies, a review, with emphasis on digestive cancer induced by Cryptosporidium parvum (Alveolata: Apicomplexa).

    Science.gov (United States)

    Benamrouz, S; Conseil, V; Creusy, C; Calderon, E; Dei-Cas, E; Certad, G

    2012-05-01

    The International Agency for Research on Cancer (IARC) identifies ten infectious agents (viruses, bacteria, parasites) able to induce cancer disease in humans. Among parasites, a carcinogenic role is currently recognized to the digenetic trematodes Schistosoma haematobium, leading to bladder cancer, and to Clonorchis sinensis or Opisthorchis viverrini, which cause cholangiocarcinoma. Furthermore, several reports suspected the potential association of other parasitic infections (due to Protozoan or Metazoan parasites) with the development of neoplastic changes in the host tissues. The present work shortly reviewed available data on the involvement of parasites in neoplastic processes in humans or animals, and especially focused on the carcinogenic power of Cryptosporidium parvum infection. On the whole, infection seems to play a crucial role in the etiology of cancer.

  8. Parasites and malignancies, a review, with emphasis on digestive cancer induced by Cryptosporidium parvum (Alveolata: Apicomplexa

    Directory of Open Access Journals (Sweden)

    Benamrouz S.

    2012-05-01

    Full Text Available The International Agency for Research on Cancer (IARC identifies ten infectious agents (viruses, bacteria, parasites able to induce cancer disease in humans. Among parasites, a carcinogenic role is currently recognized to the digenetic trematodes Schistosoma haematobium, leading to bladder cancer, and to Clonorchis sinensis or Opisthorchis viverrini, which cause cholangiocarcinoma. Furthermore, several reports suspected the potential association of other parasitic infections (due to Protozoan or Metazoan parasites with the development of neoplastic changes in the host tissues. The present work shortly reviewed available data on the involvement of parasites in neoplastic processes in humans or animals, and especially focused on the carcinogenic power of Cryptosporidium parvum infection. On the whole, infection seems to play a crucial role in the etiology of cancer.

  9. Cloning and Characterization of the Acidic Ribosomal Protein P2 of Cryptosporidium parvum, a New 17-Kilodalton Antigen▿

    Science.gov (United States)

    Priest, Jeffrey W.; Kwon, James P.; Montgomery, Joel M.; Bern, Caryn; Moss, Delynn M.; Freeman, Amanda R.; Jones, Cara C.; Arrowood, Michael J.; Won, Kimberly Y.; Lammie, Patrick J.; Gilman, Robert H.; Mead, Jan R.

    2010-01-01

    Cryptosporidium infection is commonly observed among children and immunocompromised individuals in developing countries, but large-scale outbreaks of disease among adults have not been reported. In contrast, outbreaks of cryptosporidiosis in the United States and Canada are increasingly common among patients of all ages. Thus, it seems likely that residents of regions where Cryptosporidium is highly endemic acquire some level of immunity, while residents of the developed world do not. A new immunodominant Cryptosporidium parvum antigen in the 15- to 17-kDa size range was identified as the Cryptosporidium parvum 60S acidic ribosomal protein P2 (CpP2). We developed a recombinant protein-based enzyme-linked immunosorbent assay for serologic population surveillance for antibodies that was 89% sensitive and 92% specific relative to the results of the large-format Western blot assay. The human IgG response is directed almost exclusively toward the highly conserved, carboxy-terminal 15 amino acids of the protein. Although IgG antibody cross-reactivity was documented with sera from patients with acute babesiosis, the development of an anti-CpP2 antibody response in our Peru study population correlated better with Cryptosporidium infection than with infection by any other parasitic protozoan. In Haiti, the prevalence of antibodies to CpP2 plateaus at 11 to 20 years of age. Because anti-CpP2 IgG antibodies were found only among residents of countries in the developing world where Cryptosporidium infection occurs early and often, we propose that this response may be a proxy for the intensity of infection and for acquired immunity. PMID:20410328

  10. Genome-wide upstream motif analysis of Cryptosporidium parvum genes clustered by expression profile.

    Science.gov (United States)

    Oberstaller, Jenna; Joseph, Sandeep J; Kissinger, Jessica C

    2013-07-29

    There are very few molecular genetic tools available to study the apicomplexan parasite Cryptosporidium parvum. The organism is not amenable to continuous in vitro cultivation or transfection, and purification of intracellular developmental stages in sufficient numbers for most downstream molecular applications is difficult and expensive since animal hosts are required. As such, very little is known about gene regulation in C. parvum. We have clustered whole-genome gene expression profiles generated from a previous study of seven post-infection time points of 3,281 genes to identify genes that show similar expression patterns throughout the first 72 hours of in vitro epithelial cell culture. We used the algorithms MEME, AlignACE and FIRE to identify conserved, overrepresented DNA motifs in the upstream promoter region of genes with similar expression profiles. The most overrepresented motifs were E2F (5'-TGGCGCCA-3'); G-box (5'-G.GGGG-3'); a well-documented ApiAP2 binding motif (5'-TGCAT-3'), and an unknown motif (5'-[A/C] AACTA-3'). We generated a recombinant C. parvum DNA-binding protein domain from a putative ApiAP2 transcription factor [CryptoDB: cgd8_810] and determined its binding specificity using protein-binding microarrays. We demonstrate that cgd8_810 can putatively bind the overrepresented G-box motif, implicating this ApiAP2 in the regulation of many gene clusters. Several DNA motifs were identified in the upstream sequences of gene clusters that might serve as potential cis-regulatory elements. These motifs, in concert with protein DNA binding site data, establish for the first time the beginnings of a global C. parvum gene regulatory map that will contribute to our understanding of the development of this zoonotic parasite.

  11. Triazole inhibitors of Cryptosporidium parvum inosine 5′-monophosphate dehydrogenase

    OpenAIRE

    Maurya, Sushil K.; Gollapalli, Deviprasad R.; Kirubakaran, Sivapriya; Zhang, Minjia; Johnson, Corey R.; Benjamin, Nicole N.; Hedstrom, Lizbeth; Gregory D Cuny

    2009-01-01

    Cryptosporidium parvum is an important human pathogen and potential bioterrorism agent. This protozoan parasite cannot salvage guanine or guanosine and therefore relies on inosine 5′-monophosphate dehydrogenase (IMPDH) for biosynthesis of guanine nucleotides and hence for survival. Since C. parvum IMPDH is highly divergent from the host counterpart, selective inhibitors could potentially be used to treat cryptosporidiosis with minimal effects on its mammalian host. A series of 1,2,3-triazole ...

  12. Leaching of Salmonella Senftenberg and Cryptosporidium Parvum in Intact Clay Columns

    Science.gov (United States)

    Bech, T. B.; Forslund, A.; Dalsgaard, A.; Jacobsen, O.; Jacobsen, C. S.

    2008-12-01

    Manure application on land has been associated with both environmental and public health problems, even when management is within the current guidelines. Outbreaks of infection have been associated with water or food, including processed fruits and vegetables, contaminated with animal manure. A wide range of pathogenic microorganisms can be found in animal waste, including bacteria, protozoan, and viruses. When animal waste is disposed on agricultural land different factors will influence the risk for contaminating the groundwater. 1) Animal waste application method, rate, volume and frequency will have an effect on contamination. 2) Survival of the pathogens in the soil will e.g. depend on soil water content, temperature and pH. Salmonella species can survive up to 332 days and Cryptosporidium species can remain viable for several years in the soil environment. In the present study we compared the transport between the pathogenic bacteria S. senftenberg and the pathogenic protozoan C. parvum in intact clay columns. Furthermore, we compared the effect from surface and sub-surface manure application on the transport potential. 15 intact clay columns were placed in an outdoor multi-column lysimeter for 36 days. Manure inoculated with S. senftenberg, C. parvum and chloride was added to the soil surface or injected 8 cm into the columns. Drainage water was collected from the soil columns and DNA was extracted to quantify S. senftenberg and C. parvum by quantitative PCR. In addition S. senftenberg was enumerated by plate counting. Acid yellow was applied to selected columns to visualize the pathway down through the soil column. The highest concentration of S. senftenberg was in the first drainage sample ranging from 100-10000 CFU/ml. Breakthrough curves for chloride and S. senftenberg indicates the importance of preferential flow as well as a faster transport for the bacteria compared to chloride. C. parvum is retained to a higher degree in the soil but is still found

  13. Cryptosporidium parvum genotype IIa and Giardia duodenalis assemblage A in Mytilus galloprovincialis on sale at local food markets.

    Science.gov (United States)

    Giangaspero, Annunziata; Papini, Roberto; Marangi, Marianna; Koehler, Anson V; Gasser, Robin B

    2014-02-03

    To date, there has been no study to establish the genotypic or subgenotypic identities of Cryptosporidium and Giardia in edible shellfish. Here, we explored the genetic composition of these protists in Mytilus galloprovincialis (Mediterranean mussel) purchased from three markets in the city of Foggia, Italy, from May to December 2012. Samples from the digestive glands, gills and haemolymph were tested by nested PCR, targeting DNA regions within the 60 kDa glycoprotein (gp60) gene of Cryptosporidium, and the triose-phosphate isomerase (tpi) and β-giardin genes of Giardia. In total, Cryptosporidium and Giardia were detected in 66.7% of mussels (M. galloprovincialis) tested. Cryptosporidium was detected mostly between May and September 2012. Sequencing of amplicons showed that 60% of mussels contained Cryptosporidium parvum genotype IIa (including subgenotypes A15G2R1, IIaA15G2 and IIaA14G3R1), 23.3% Giardia duodenalis assemblage A, and 6.6% had both genetic types. This is the first report of these types in fresh, edible shellfish, particularly the very commonly consumed M. galloprovincialis from highly frequented fish markets. These genetic types of Cryptosporidium and Giardia are known to infect humans and thus likely to represent a significant public health risk. The poor observance of hygiene rules by vendors, coupled to the large numbers of M. galloprovincialis sold and the eating habits of consumers in Italy, call for more effective sanitary measures pertaining to the selling of fresh shellfish in street markets.

  14. Cryptosporidium parvum GP60 subtypes in dairy cattle from Buenos Aires, Argentina

    Science.gov (United States)

    Cryptosporidium parvum from 73 dairy calves less than two months old from Buenos Aires province (Argentina) were molecularly characterized using sequence analysis of the GP60 gene. Seventy five sequences were obtained, and seven different subtypes were identified, all belonging to the IIa subtype f...

  15. Wastewater treatment with Moringa oleifera seed extract: Impact on turbidity and sedimentation of Cryptosporidium parvum oocysts

    DEFF Research Database (Denmark)

    Petersen, Heidi H.; Woolsey, Ian; Dalsgaard, Anders

    produced from seeds of the Moringa oleifera tree (MO) in reducing Cryptosporidium parvum oocysts and turbidity in wastewater. To a total of 5 x 12 glass jars containing 500 ml wastewater samples from a Danish treatment plant, 1.2 x 106 ± 1.2 x 105 oocysts L-1 were added. To half of the wastewater samples 8...

  16. Effects of Surfactants on Cryptosporidium parvum Mobility in Agricultural Soils from Illinois and Utah

    Science.gov (United States)

    Darnault, C. J.; Koken, E.; Jacobson, A. R.; Powelson, D.

    2011-12-01

    The occurence of the parasitic protozoan Cryptosporidium parvum in rural and agricultural watersheds due to agricultural activities and wildlife is inevitable. Understanding the behavior of C. parvum oocysts in the environment is critical for the protection of public health and the environment. To better understand the mechanisms by which the pathogen moves through soils and contaminates water resources, we study their mobility under conditions representative of real-world scenarios, where both C. parvum and chemicals that affect their fate are present in soils. Surfactants occur widely in soils due to agricultural practices such as wastewater irrigation and the application of pesticides or soil wetting agents. They affect water tension and, consequently, soil infiltration processes and the air-water interfaces in soil pores where C. parvum may be retained. We investigate the effects of surfactants on the mobility of C. parvum oocysts in agricultural soils from Illinois and Utah under unsaturated flow conditions. As it is critical to examine C. parvum in natural settings, we also developed a quantification method using RT-PCR for monitoring C. parvum oocysts in environmental soil and water samples. We optimized physico-chemical parameters to disrupt C. parvum oocysts and extract their DNA, and developed isolation methods to separate C. parvum oocysts from colloids in natural soil samples. The results of this research will lead to the development of an accurate and sensitive molecular method for the monitoring of C. parvum oocysts in environmental soil and water samples, and will further our understanding of the mechanisms controlling the behavior of C. parvum oocysts in soils, in particular the role of vadose zone processes, sorption to soil and surfactants.

  17. Inactivation of Cryptosporidium parvum oocysts using medium- and low-pressure ultraviolet radiation.

    Science.gov (United States)

    Craik, S A; Weldon, D; Finch, G R; Bolton, J R; Belosevic, M

    2001-04-01

    The effect of ultraviolet radiation from low- and medium-pressure mercury arc lamps on Cryptosporidium parvum oocysts was studied using a collimated beam apparatus. Experiments were conducted using parasites suspended in both filtered surface water and phosphate buffered laboratory water. Inactivation of oocysts was measured as reduction in infectivity using a CD-1 neonatal mouse model and was found to be a non-linear function of UV dose over the range of germicidal doses tested (0.8-119 mJ/cm2). Oocyst inactivation increased rapidly with UV dose at doses less than 25 mJ/cm2 with two and three log-units inactivation at approximately 10 and 25 mJ/cm2, respectively. The cause of significant leveling-off and tailing in the UV inactivation curve at higher doses was not determined. Maximum measured oocyst inactivation ranged from 3.4 to greater than 4.9 log-units and was dependent on different batches of parasites. Water type and temperature, the concentration of oocysts in the suspension, and the UV irradiance did not have significant impacts on oocyst inactivation. When compared on the basis of germicidal UV dose, the oocysts were equally sensitive to low- and medium-pressure UV radiation. With respect to Cryptosporidium, both low- and medium-pressure ultraviolet radiation are attractive alternatives to conventional chemical disinfection methods in drinking water treatment.

  18. (31)P NMR of apicomplexans and the effects of risedronate on Cryptosporidium parvum growth.

    Science.gov (United States)

    Moreno, B; Bailey, B N; Luo, S; Martin, M B; Kuhlenschmidt, M; Moreno, S N; Docampo, R; Oldfield, E

    2001-06-15

    High-resolution 303.6 MHz (31)P NMR spectra have been obtained of perchloric acid extracts of Plasmodium berghei trophozoites, Toxoplasma gondii tachyzoites, and Cryptosporidium parvum oocysts. Essentially complete resonance assignments have been made based on chemical shifts and by coaddition of authentic reference compounds. Signals corresponding to inorganic pyrophosphate were detected in all three species. In T. gondii and C. parvum, additional resonances were observed corresponding to linear triphosphate as well as longer chain polyphosphates. Spectra of P. berghei and T. gondii also indicated the presence of phosphomonoesters and nucleotide phosphates. We also report that the pyrophosphate analog drug, risedronate (used in bone resorption therapy), inhibits the growth of C. parvum in a mouse xenograft model. When taken together, our results indicate that all the major disease-causing apicomplexan parasites contain extensive stores of condensed phosphates and that as with Plasmodium falciparum and T. gondii, the pyrophosphate analog drug risedronate is an inhibitor of C. parvum cell growth.

  19. Cloning and expression of gene encoding P23 protein from Cryptosporidium parvum

    Directory of Open Access Journals (Sweden)

    Dinh Thi Bich Lan

    2014-12-01

    Full Text Available We cloned the cp23 gene coding P23 (glycoprotein from Cryptosporidium parvum isolated from Thua Thien Hue province, Vietnam. The coding region of cp23 gene from C. parvum is 99% similar with cp23 gene deposited in NCBI (accession number: U34390. SDS-PAGE and Western blot analysis showed that the cp23 gene in E. coli BL21 StarTM (DE3 produced polypeptides with molecular weights of approximately 37, 40 and 49 kDa. These molecules may be non-glycosylated or glycosylated P23 fusion polypeptides. Recombinant P23 protein purified by GST (glutathione S-transferase affinity chromatography can be used as an antigen for C. parvum antibody production as well as to develop diagnostic kit for C. parvum.

  20. MODELING CRYPTOSPORIDIUM PARVUM OOCYST INACTIVATION AND BROMATE IN A FLOW-THROUGH OZONE CONTACTOR TREATING NATURAL WATER

    Science.gov (United States)

    A reactive transport model was developed to simultaneously predict Cryptosporidium parvum oocyst inactivation and bromate formation during ozonation of natural water. A mechanistic model previously established to predict bromate formation in organic-free synthetic waters w...

  1. Intracellular levels of the viral symbiont CPV in Cryptosporidium parvum correlate with fecundity of the parasite in dairy calves

    Science.gov (United States)

    Previous reports have cited differences in clinical signs and oocyst output among strains of Cryptosporidium parvum. The purpose of this study was to determine if levels of the C. parvum intracellular viral symbiont CPV correlated with observed clinical and parasitological differences. Calves infe...

  2. Removal and fate of Cryptosporidium parvum, Clostridium perfringens and small-sized centric diatoms (Stephanodiscus hantzschii) in slow sand filters

    NARCIS (Netherlands)

    Hijnen, W.A.M.; Dullemont, Y.J.; Schijven, J.F.; Brouwer-Hanzens, Anke J.; Rosielle, M.; Medema, Gerriet Jan

    2007-01-01

    The decimal elimination capacity (DEC) of slow sand filtration (SSF) for Cryptosporidium parvum was assessed to enable quantitative microbial risk analysis of a drinking water production plant. A mature pilot plant filter of 2.56 m2 was loaded with C. parvum oocysts and two other persistent organism

  3. Molecular detection of Cryptosporidium spp. infections in water buffaloes from northeast Thailand.

    Science.gov (United States)

    Inpankaew, Tawin; Jiyipong, Tawisa; Wongpanit, Kannika; Pinyopanuwat, Nongnuch; Chimnoi, Wissanuwat; Kengradomkij, Chanya; Xuan, Xuenan; Igarashi, Ikuo; Xiao, Lihua; Jittapalapong, Sathaporn

    2014-02-01

    The objectives of this study were to determine the individual and herd-level prevalence and genotype of Cryptosporidium and to identify putative risk factors associated with Cryptosporidium spp. infections in water buffaloes in northeast Thailand. Fecal samples from 600 water buffaloes of 287 farms in six provinces were collected and tested using DMSO-modified acid-fast staining and polymerase chain reaction. The overall prevalence of Cryptosporidium infections in buffaloes was 5.7 and 8.7% among individual animals and herds, respectively. The provinces with highest infected Cryptosporidium were located in the Sakon Nakhon Basin in the northern part of the region. In addition, higher herd prevalence was observed among farms with more than five buffaloes (30%) than those with five or less animals (16.2%). Thirty (88.2%) of the 34 Cryptosporidium-positive samples were Cryptosporidium parvum and four (11.8%) were Cryptosporidium ryanae.

  4. Epidemiology of Cryptosporidium infection in cattle in China: a review

    Directory of Open Access Journals (Sweden)

    Gong Chao

    2017-01-01

    Full Text Available The present review discusses the findings of cryptosporidiosis research conducted in cattle in China and highlights the currently available information on Cryptosporidium epidemiology, genetic diversity, and distribution in China, which is critical to understanding the economic and public health importance of cryptosporidiosis transmission in cattle. To date, 10 Cryptosporidium species have been detected in cattle in China, with an overall infection rate of 11.9%. The highest rate of infection (19.5% was observed in preweaned calves, followed by that in juveniles (10.69%, postweaned juveniles (9.0%, and adult cattle (4.94%. The dominant species were C. parvum in preweaned calves and C. andersoni in postweaned, juvenile, and adult cattle. Zoonotic Cryptosporidium species (C. parvum and C. hominis were found in cattle, indicating the possibility of transmission between humans and cattle. Different cattle breeds had significant differences in the prevalence rate and species of Cryptosporidium. This review demonstrates an age-associated, breed-associated, and geographic-related occurrence of Cryptosporidium and provides references for further understanding of the epidemiological characteristics, and for preventing and controlling the disease.

  5. Epidemiology of Cryptosporidium infection in cattle in China: a review

    Science.gov (United States)

    Gong, Chao; Cao, Xue-Feng; Deng, Lei; Li, Wei; Huang, Xiang-Ming; Lan, Jing-Chao; Xiao, Qi-Cheng; Zhong, Zhi-Jun; Feng, Fan; Zhang, Yue; Wang, Wen-Bo; Guo, Ping; Wu, Kong-Ju; Peng, Guang-Neng

    2017-01-01

    The present review discusses the findings of cryptosporidiosis research conducted in cattle in China and highlights the currently available information on Cryptosporidium epidemiology, genetic diversity, and distribution in China, which is critical to understanding the economic and public health importance of cryptosporidiosis transmission in cattle. To date, 10 Cryptosporidium species have been detected in cattle in China, with an overall infection rate of 11.9%. The highest rate of infection (19.5%) was observed in preweaned calves, followed by that in juveniles (10.69%), postweaned juveniles (9.0%), and adult cattle (4.94%). The dominant species were C. parvum in preweaned calves and C. andersoni in postweaned, juvenile, and adult cattle. Zoonotic Cryptosporidium species (C. parvum and C. hominis) were found in cattle, indicating the possibility of transmission between humans and cattle. Different cattle breeds had significant differences in the prevalence rate and species of Cryptosporidium. This review demonstrates an age-associated, breed-associated, and geographic-related occurrence of Cryptosporidium and provides references for further understanding of the epidemiological characteristics, and for preventing and controlling the disease. PMID:28098070

  6. Biofilm roughness determines Cryptosporidium parvum retention in environmental biofilms.

    Science.gov (United States)

    DiCesare, E A Wolyniak; Hargreaves, B R; Jellison, K L

    2012-06-01

    The genus Cryptosporidium is a group of waterborne protozoan parasites that have been implicated in significant outbreaks of gastrointestinal infections throughout the world. Biofilms trap these pathogens and can contaminate water supplies through subsequent release. Biofilm microbial assemblages were collected seasonally from three streams in eastern Pennsylvania and used to grow biofilms in laboratory microcosms. Daily oocyst counts in the influx and efflux flow allowed the calculation of daily oocyst retention in the biofilm. Following the removal of oocysts from the influx water, oocyst attachment to the biofilm declined to an equilibrium state within 5 days that was sustained for at least 25 days. Varying the oocyst loading rate for the system showed that biofilm retention could be saturated, suggesting that discrete binding sites determined the maximum number of oocysts retained. Oocyst retention varied seasonally but was consistent across all three sites; however, seasonal oocyst retention was not consistent across years at the same site. No correlation between oocyst attachment and any measured water quality parameter was found. However, oocyst retention was strongly correlated with biofilm surface roughness and roughness varied among seasons and across years. We hypothesize that biofilm roughness and oocyst retention are dependent on environmentally driven changes in the biofilm community rather than directly on water quality conditions. It is important to understand oocyst transport dynamics to reduce risks of human infection. Better understanding of factors controlling biofilm retention of oocysts should improve our understanding of oocyst transport at different scales.

  7. Seven years' experience with Cryptosporidium parvum in Guinea-Bissau, West Africa

    DEFF Research Database (Denmark)

    Perch, M; Sodemann, Morten; Jakobsen, M S;

    2001-01-01

    , exceeded only by Giardia lamblia which was found in 14.8% of the samples. The highest prevalence of cryptosporidium was found in children aged 6-11 months, whereas the prevalence of other enteric parasites increased with age. Cryptosporidiosis showed a marked seasonal variation, with peak prevalences found...... consistently at the beginning of or just before the rainy seasons, May through July. By contrast, no seasonality was found for the enteric parasites Giardia lamblia or Entamoeba histolytica. We conclude that Cryptosporidium parvum is an important pathogen in children with diarrhoea....

  8. Seven years' experience with Cryptosporidium parvum in Guinea-Bissau, West Africa

    DEFF Research Database (Denmark)

    Perch, M; Sodemann, Morten; Jakobsen, M S

    2001-01-01

    , exceeded only by Giardia lamblia which was found in 14.8% of the samples. The highest prevalence of cryptosporidium was found in children aged 6-11 months, whereas the prevalence of other enteric parasites increased with age. Cryptosporidiosis showed a marked seasonal variation, with peak prevalences found...... consistently at the beginning of or just before the rainy seasons, May through July. By contrast, no seasonality was found for the enteric parasites Giardia lamblia or Entamoeba histolytica. We conclude that Cryptosporidium parvum is an important pathogen in children with diarrhoea....

  9. Effect of chlorine, blanching, freezing, and microwave heating on Cryptosporidium parvum viability inoculated on green peppers.

    Science.gov (United States)

    Duhain, G L M C; Minnaar, A; Buys, E M

    2012-05-01

    Cryptosporidium parvum oocysts have been found on the surface of vegetables in both developed and developing countries. C. parvum can contaminate vegetables via various routes, including irrigation water. This study investigated the effect of individual treatments of chlorine, blanching, blast freezing, and microwave heating, as well as combined treatments of chlorine and freezing, and chlorine and microwave heating on the viability of C. parvum oocysts inoculated on green peppers. The viability of the oocysts after the treatments was assessed using propidium iodide and a flow cytometer. Based on the propidium iodide staining, the chlorine treatments did not affect the viability of the oocysts. Blast freezing significantly inactivated 20% of the oocysts. Microwave heating and blanching significantly inactivated 93% of oocysts. Treatment with chlorine followed by blast freezing did not affect the viability of the oocysts significantly. Treatment with chlorine and microwave heating was significantly more effective than microwave heating alone and inactivated 98% of the oocysts. The study indicates that C. parvum oocysts are sensitive to heat and, to some extent, to blast freezing, but are resistant to chlorine. Therefore, the use of chlorine during vegetable processing is not a critical control point for C. parvum oocysts, and the consumption of raw or minimally processed vegetables may constitute a health risk as C. parvum oocysts can still be found viable on ready-to-eat, minimally processed vegetables.

  10. Detection of Cryptosporidium parvum Oocysts on Fresh Produce Using DNA Aptamers.

    Directory of Open Access Journals (Sweden)

    Asma Iqbal

    Full Text Available There are currently no standard methods for the detection of Cryptosporidium spp., or other protozoan parasites, in foods, and existing methods are often inadequate, with low and variable recovery efficiencies. Food testing is difficult due to the low concentrations of parasites, the difficulty in eluting parasites from some foods, the lack of enrichment methods, and the presence of PCR inhibitors. The main objectives of the present study were to obtain DNA aptamers binding to the oocyst wall of C. parvum, and to use the aptamers to detect the presence of this parasite in foods. DNA aptamers were selected against C. parvum oocysts using SELEX (Systematic Evolution of Ligands by EXponential enrichment. Ten rounds of selection led to the discovery of 14 aptamer clones with high affinities for C. parvum oocysts. For detecting parasite-bound aptamers, a simple electrochemical sensor was employed, which used a gold nanoparticle-modified screen-printed carbon electrode. This aptasensor was fabricated by self-assembling a hybrid of a thiolated ssDNA primer and the anti- C. parvum aptamer. Square wave voltammetry was employed to quantitate C. parvum in the range of 150 to 800 oocysts, with a detection limit of approximately 100 oocysts. The high sensitivity and specificity of the developed aptasensor suggests that this novel method is very promising for the detection and identification of C. parvum oocysts on spiked fresh fruits, as compared to conventional methods such as microscopy and PCR.

  11. Cryptosporidium parvum (Apicomplexa: Cryptosporidiidae) oocyst and sporozoite antigens recognized by bovine colostral antibodies.

    OpenAIRE

    Tilley, M; Fayer, R; Guidry, A; Upton, S J; Blagburn, B. L.

    1990-01-01

    Colostral whey from seven hyperimmunized and two control cows (hyperimmune bovine colostrum) was examined by Western immunoblotting for the presence of antibody against oocysts and sporozoites of Cryptosporidium parvum, using rabbit anti-bovine immunoglobulin A (IgA), IgG1, IgG2, and IgM antibodies, followed by a horseradish peroxidase goat anti-rabbit polyvalent antibody. Although considerable variation was found in binding activity between cows on different immunization protocols, IgA and I...

  12. Inactivation of Cryptosporidium parvum Oocysts in Fresh Apple Cider by UV Irradiation

    Science.gov (United States)

    Hanes, D. E.; Worobo, R. W.; Orlandi, P. A.; Burr, D. H.; Miliotis, M. D.; Robl, M. G.; Bier, J. W.; Arrowood, M. J.; Churey, J. J.; Jackson, G. J.

    2002-01-01

    This study evaluated the efficacy of UV irradiation on the inactivation of Cryptosporidium parvum oocysts in fresh apple cider. Cider was inoculated with oocysts and exposed to 14.32 mJ of UV irradiation/cm2. Oocyst viability was assessed with the gamma interferon gene knockout (GKO) mouse and infant BALB/cByJ mouse models. All GKO mice challenged with UV-treated cider demonstrated no morbidity or mortality, and infant BALB/c mice challenged with treated cider were negative for the presence of C. parvum. In contrast, the GKO mice challenged with non-UV-treated inoculated cider died and the parasite was detected in the ileums of all challenged infant mice. This study shows that UV irradiation can be used to inactivate C. parvum in fresh apple cider. PMID:12147528

  13. CP2 gene as a useful viability marker for Cryptosporidium parvum.

    Science.gov (United States)

    Lee, Soo-Ung; Joung, Migyo; Ahn, Myoung-Hee; Huh, Sun; Song, Hyunje; Park, Woo-Yoon; Yu, Jae-Ran

    2008-02-01

    The validity of the CP2 gene of Cryptosporidium parvum as a viability marker was evaluated using absolute quantitative real-time polymerase chain reaction (qPCR) assays. Total ribonucleic acid (RNA) was isolated from live and heat-killed C. parvum oocysts, and complementary deoxyribonucleic acid was synthesized and used as a template. The most accurate number of viable C. parvum oocysts was predicted when the CP2 gene was used as a target gene. The lower detection limit of the CP2 gene was ten oocysts, which was the most sensitive among examined target genes. With heat shock induction, only hsp70 messenger RNA (mRNA) was induced, and the predicted viable oocyst number was increased by heat shock for this marker. The CP2, hsp70, Cryptosporidium oocyst wall protein, and beta-tubulin mRNAs were not detected in heat-killed oocysts, but the 18S ribosomal ribonucleic acid (rRNA) showed heat stability until 48 h after heat killing. Although the 18S rRNA demonstrated the fastest response in crossing point (CP) value among the examined primer sets in qPCR, overestimation of viable oocysts was noted in the analysis with this gene. In conclusion, the CP2 gene was identified as the most sensitive, reliable, and accurate candidate of a viability marker of C. parvum by qPCR evaluation.

  14. Multicenter Evaluation of BD Max Enteric Parasite Real-Time PCR Assay for Detection of Giardia duodenalis, Cryptosporidium hominis, Cryptosporidium parvum, and Entamoeba histolytica

    Science.gov (United States)

    Relich, R. F.; Doyle, L.; Espina, N.; Fuller, D.; Karchmer, T.; Lainesse, A.; Mortensen, J. E.; Pancholi, P.; Veros, W.; Harrington, S. M.

    2016-01-01

    Common causes of chronic diarrhea among travelers worldwide include protozoan parasites. The majority of parasitic infections are caused by Giardia duodenalis, Entamoeba histolytica, Cryptosporidium parvum, and Cryptosporidium hominis. Similarly, these species cause the majority of parasitic diarrhea acquired in the United States. Detection of parasites by gold standard microscopic methods is time-consuming and requires considerable expertise; enzyme immunoassays and direct fluorescent-antibody (DFA) stains have lowered hands-on time for testing, but improvements in sensitivity and technical time may be possible with a PCR assay. We performed a clinical evaluation of a multiplex PCR panel, the enteric parasite panel (EPP), for the detection of these common parasites using the BD Max instrument, which performs automated extraction and amplification. A total of 2,495 compliant specimens were enrolled, including 2,104 (84%) specimens collected prospectively and 391 (16%) specimens collected retrospectively. Approximately equal numbers were received in 10% formalin (1,273 specimens) and unpreserved (1,222 specimens). The results from the EPP were compared to those from alternate PCR and bidirectional sequencing (APCR), as well as DFA (G. duodenalis and C. parvum or C. hominis) or trichrome stain (E. histolytica). The sensitivity and specificity for prospective and retrospective specimens combined were 98.2% and 99.5% for G. duodenalis, 95.5% and 99.6 for C. parvum or C. hominis, and 100% and 100% for E. histolytica, respectively. The performance of the FDA-approved BD Max EPP compared well to the reference methods and may be an appropriate substitute for microscopic examination or immunoassays. PMID:27535690

  15. A novel Cryptosporidium parvum antigen, CP2, preferentially associates with membranous structures.

    Science.gov (United States)

    O'Hara, Steven P; Yu, Jae-Ran; Lin, Jim Jung-Ching

    2004-03-01

    The present study addresses the cloning and characterization of a Cryptosporidium parvum antigen, CP2. Sequencing of cDNA and genomic clones revealed a novel gene capable of coding a message of 2,136 nucleotides flanked by 28 and 140 nucleotides of the 5'- and 3'-noncoding regions, respectively. The deduced amino acid sequence suggests that CP2 is a secreted and/or membrane protein. Immunofluorescence microscopy detected CP2 enrichment in sporozoites that subsequently appeared to encase type I meronts in infected HCT-8 cells. Immunogold electron microscopy revealed that CP2 consistently localized to membranous structures throughout development. In addition, progression from macrogametocyte to sporulated oocyst revealed CP2 initially at the periphery of amylopectin-like granules, in the cytoplasm and discrete vesicles, the parasitophorous vacuole, on the surface of sporozoites, and finally on the parasitophorous vacuole membrane (PVM). The observed expression pattern suggests that CP2 may be involved in the invasion process and/or PVM integrity.

  16. Calf-level risk factors for neonatal diarrhea and shedding of Cryptosporidium parvum in Ontario dairy calves.

    Science.gov (United States)

    Trotz-Williams, Lise A; Wayne Martin, S; Leslie, Kenneth E; Duffield, Todd; Nydam, Daryl V; Peregrine, Andrew S

    2007-11-15

    This work was conducted to investigate calf-level factors that influence the risk of neonatal diarrhea and shedding of Cryptosporidium parvum oocysts in calves, on dairy farms in Ontario with histories of calf diarrhea or cryptosporidiosis. Fecal samples were collected weekly for 4 weeks from each of 1045 calves under 30 days of age on 11 dairy farms in south-western Ontario during the summer of 2003 and the winter of 2004. A questionnaire designed to gather information on calf-level management factors was administered on farm for each calf in the study. Samples were examined for C. parvum oocysts by microscopy, and a subset of specimens was also tested for enterotoxigenic Escherichia coli, Salmonella, bovine rotavirus and bovine coronavirus. The consistency of each sample was scored and recorded at the time of collection in order to assess the presence or absence of diarrhea. In addition, a blood sample was taken from each calf upon enrollment in the study, for assessment of maternal antibody transfer and for polymerase chain reaction testing for persistent bovine viral diarrhea virus infection. Using the GLLAMM function in Stata 9.0, multilevel regression techniques were employed to investigate associations between management practices and the risk of C. parvum shedding or diarrhea. C. parvum oocysts were detected in the feces of 78% of the 919 calves from which all four fecal samples had been collected. Furthermore, 73% of the 846 calves for which all four fecal consistency scores had been recorded were diarrheic at the time of collection of at least one sample. Significant predictors of the calf-level risk of C. parvum shedding included the use of calf diarrhea prophylaxis in pregnant cows, and the type of maternity facilities in which the calves were born. Factors associated with an increased risk of diarrhea were leaving the calf with the dam for more than an hour after birth, and the birth of a calf in the summer as opposed to winter. Calves shedding C

  17. Identification of putative cis-regulatory elements in Cryptosporidium parvum by de novo pattern finding

    Directory of Open Access Journals (Sweden)

    Kissinger Jessica C

    2007-01-01

    Full Text Available Abstract Background Cryptosporidium parvum is a unicellular eukaryote in the phylum Apicomplexa. It is an obligate intracellular parasite that causes diarrhea and is a significant AIDS-related pathogen. Cryptosporidium parvum is not amenable to long-term laboratory cultivation or classical molecular genetic analysis. The parasite exhibits a complex life cycle, a broad host range, and fundamental mechanisms of gene regulation remain unknown. We have used data from the recently sequenced genome of this organism to uncover clues about gene regulation in C. parvum. We have applied two pattern finding algorithms MEME and AlignACE to identify conserved, over-represented motifs in the 5' upstream regions of genes in C. parvum. To support our findings, we have established comparative real-time -PCR expression profiles for the groups of genes examined computationally. Results We find that groups of genes that share a function or belong to a common pathway share upstream motifs. Different motifs are conserved upstream of different groups of genes. Comparative real-time PCR studies show co-expression of genes within each group (in sub-sets during the life cycle of the parasite, suggesting co-regulation of these genes may be driven by the use of conserved upstream motifs. Conclusion This is one of the first attempts to characterize cis-regulatory elements in the absence of any previously characterized elements and with very limited expression data (seven genes only. Using de novo pattern finding algorithms, we have identified specific DNA motifs that are conserved upstream of genes belonging to the same metabolic pathway or gene family. We have demonstrated the co-expression of these genes (often in subsets using comparative real-time-PCR experiments thus establishing evidence for these conserved motifs as putative cis-regulatory elements. Given the lack of prior information concerning expression patterns and organization of promoters in C. parvum we

  18. Multiplex-Touchdown PCR to Simultaneously Detect Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the Major Causes of Traveler's Diarrhea.

    Science.gov (United States)

    Shin, Ji-Hun; Lee, Sang-Eun; Kim, Tong Soo; Ma, Da-Won; Chai, Jong-Yil; Shin, Eun-Hee

    2016-10-01

    This study aimed to develop a multiplex-touchdown PCR method to simultaneously detect 3 species of protozoan parasites, i.e., Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the major causes of traveler's diarrhea and are resistant to standard antimicrobial treatments. The target genes included the Cryptosporidium oocyst wall protein for C. parvum, Glutamate dehydrogenase for G. lamblia, and 18S ribosomal RNA (18S rRNA) for C. cayetanensis. The sizes of the amplified fragments were 555, 188, and 400 bps, respectively. The multiplex-touchdown PCR protocol using a primer mixture simultaneously detected protozoa in human stools, and the amplified gene was detected in >1×10(3) oocysts for C. parvum, >1×10(4) cysts for G. lamblia, and >1 copy of the 18S rRNA gene for C. cayetanensis. Taken together, our protocol convincingly demonstrated the ability to simultaneously detect C. parvum, G. lamblia, and C. cayetanenesis in stool samples.

  19. Prevalence of Cryptosporidium parvum in private drinking water cisterns in Bani-Kenanah district, northern Jordan.

    Science.gov (United States)

    Abo-Shehada, Mahmoud; Hindyia, Mona; Saiah, Abbass

    2004-10-01

    Due to water scarcity in Jordan, the water authority only pump the water once or twice a week to the population. Thus people in rural areas, including the Bani-Kenanah district, make the most of their water resources by storing rainwater in private reservoirs for use during periods of water shortage. These reservoirs include; underground cisterns and concrete or metal tanks. The water collected in these reservoirs is at risk of contamination. During the period March-July 2002, the three types of reservoirs from 368 households were surveyed for presence of Escherichia coli and Cryptosporidium parvum, indicators of contamination. The cistern was the most contaminated reservoir with 17% (95% CI: 13,22) for E. coli (significant, P<0.05), and 2% (95% CI: 1,4) for C. parvum. Only 1% (95% CI: 1,6) of the metal reservoirs had E. coli, while concrete reservoirs were free. No C. parvum oocysts were detected in either the concrete or metal reservoirs. Reservoirs opening at floor level and the bucket kept outside the reservoir were significant (P<0.05) enhancing risk factors for contamination with C. parvum.

  20. Cell culture-Taqman PCR assay for evaluation of Cryptosporidium parvum disinfection.

    Science.gov (United States)

    Keegan, Alexandra R; Fanok, Stella; Monis, Paul T; Saint, Christopher P

    2003-05-01

    Cryptosporidium parvum represents a challenge to the water industry and a threat to public health. In this study, we developed a cell culture-quantitative PCR assay to evaluate the inactivation of C. parvum with disinfectants. The assay was validated by using a range of disinfectants in common use in the water industry, including low-pressure UV light (LP-UV), ozone, mixed oxidants (MIOX), and chlorine. The assay was demonstrated to be reliable and sensitive, with a lower detection limit of a single infectious oocyst. Effective oocyst inactivation was achieved (>2 log(10) units) with LP-UV (20 mJ/cm(2)) or 2 mg of ozone/liter (for 10 min). MIOX and chlorine treatments of oocysts resulted in minimal effective disinfection, with disinfection systems for drinking water and recycled water.

  1. Cryptosporidium parvum (Apicomplexa: Cryptosporidiidae) oocyst and sporozoite antigens recognized by bovine colostral antibodies.

    Science.gov (United States)

    Tilley, M; Fayer, R; Guidry, A; Upton, S J; Blagburn, B L

    1990-09-01

    Colostral whey from seven hyperimmunized and two control cows (hyperimmune bovine colostrum) was examined by Western immunoblotting for the presence of antibody against oocysts and sporozoites of Cryptosporidium parvum, using rabbit anti-bovine immunoglobulin A (IgA), IgG1, IgG2, and IgM antibodies, followed by a horseradish peroxidase goat anti-rabbit polyvalent antibody. Although considerable variation was found in binding activity between cows on different immunization protocols, IgA and IgG1 in whey recognized a greater variety of C. parvum antigens than did IgG2 and IgM. A band at 9 to 10 kilodaltons appeared unique in that it was recognized only by IgA.

  2. Viability and fate of Cryptosporidium parvum and Giardia lamblia in tubular anaerobic digesters.

    Science.gov (United States)

    Kinyua, Maureen N; Trimmer, John; Izurieta, Ricardo; Cunningham, Jeffrey; Ergas, Sarina J

    2016-06-01

    In many developing countries where pathogenic diseases of animal waste origin, such as giardiasis and cryptosporidiosis, are often prevalent, facilities are limited to treat livestock waste. However, household-scale anaerobic digesters are currently being promoted for bioenergy production from livestock manure. Since the effluent is often used as a fertilizer for food crops, it is critical to understand the effect of environmental conditions within household-scale digesters on the viability of Cryptosporidium parvum oocysts and Giardia lamblia cysts. In this study, key environmental parameters affecting (oo)cyst inactivation were measured in four tubular anaerobic digesters, which are a type of household-scale digester promoted for treatment of swine waste in rural Costa Rica. Interviews and participant observations were used to understand digester operation and maintenance procedures. Ambient temperatures (21-24°C), near-neutral pH, total ammonia nitrogen (TAN) concentrationsGiardia lamblia and Cryptosporidium parvum were 0.155 ± 0.041 and 0.054 ± 0.006 day(-1), respectively. Temperature and volatile fatty acids were the main factors contributing to Cryptosporidium parvum and Giardia lamblia inactivation. A mathematical model was developed that predicts the concentration of (oo)cysts in the liquid effluent of tubular digesters like those observed in Costa Rica. A mathematical model was developed that predicts the concentration of (oo)cysts in the liquid effluent of tubular digesters like those observed in Costa Rica. Two dimensionless groups can be used to predict the performance of the digesters for inactivating pathogens; both dimensionless groups depend upon the average HRT in the digester. This is the first study to combine mathematical modeling with qualitative analysis, field and laboratory studies to predict the concentrations of (oo)cysts in tubular digester effluents.

  3. Application of an ELISA-type screen printed electrode-based potentiometric assay to the detection of Cryptosporidium parvum oocysts.

    Science.gov (United States)

    Laczka, Olivier; Skillman, Lucy; Ditcham, William G; Hamdorf, Brenton; Wong, Danny K Y; Bergquist, Peter; Sunna, Anwar

    2013-11-01

    We report a novel electrochemical method for the rapid detection of the parasitic protozoan, Cryptosporidium parvum. An antibody-based capture format was transferred onto screen-printed electrodes and the presence of horseradish peroxidase-labelled antibodies binding to the oocysts was potentiometrically detected. This method allowed the detection of 5 × 10(2)Cryptosporidium oocysts per mL in 60 min.

  4. Fluorescent in situ hybridization as a tool to retrospectively identify Cryptosporidium parvum and Giardia lamblia in samples from terrestrial mammalian wildlife.

    Science.gov (United States)

    Bednarska, Malgorzata; Bajer, Anna; Sinski, Edward; Girouard, Autumn S; Tamang, Leena; Graczyk, Thaddeus K

    2007-02-01

    Fecal samples of five terrestrial mammalian wildlife species stored at 4 degrees C or at -20 degrees C for up to 36 months have been tested for human zoonotic enteric parasites (i.e., Cryptosporidium parvum and Giardia lamblia) using combined fluorescent in situ hybridization (FISH) and direct fluorescent antibody techniques. The prevalence of C. parvum and G. lamblia varied from 20 to 63% (mean, 45.8%) and from 13 to 100% (mean, 53.2%), respectively. The prevalence of C. parvum and G. lamblia infections was higher in small rodents (mean, 68.5%) than in other wildlife (mean, 21%). Overall, 31.1% of animals were coinfected, and coinfections were more prevalent in small rodents (mean, 52%) than in other wildlife species (mean, 13.2%). The present study has shown that the FISH assay can be retrospectively applied to fecal samples for the identification of C. parvum oocysts, but is less suitable for the identification of G. lamblia cysts in such samples. Terrestrial mammalian wildlife, particularly small rodents, can contribute to watershed contamination with C. parvum oocysts and G. lamblia cysts. To control contamination, the management of pristine watersheds used for drinking water purposes should incorporate control measures for terrestrial wildlife, especially field rodents residing within such watersheds.

  5. Low-level detection of Cryptosporidium parvum in field water using optical microfluidic biosensors

    Science.gov (United States)

    Angus, Scott V.; Kwon, Hyuck-Jin; Yoon, Jeong-Yeol

    2012-03-01

    Cryptosporidium parvum is a difficult-to-detect protozoan that causes diarrhea in the healthy adults and death in immunocompromised individuals. While it is easy to understand the transmission routes of Cryptosporidium, it is currently difficult to identify low concentrations of Cryptosporidium, especially when following EPA method 1623, which can easily require tens of liters of water to get a positive signal. The current detection method is unacceptable and severely inefficient when taking into account the time that goes into concentrating a sample, actual assays, and training associated with the assays. Using our method, it is possible to use only 15 μL of sample, which is an immunoagglutination assay that uses Mie scatter intensity changes to detect different Cryptosporidium concentrations. In addition to creating a standard curve using a clean sample matrix (i.e., phosphate buffered saline), field samples were collected from a chlorine treated swimming pool, a sump located on a farm, and a turtle pond. Each sample had different intensity changes but the trend represented within the data was the same. This assay has a detection limit of 100-101 oocysts/mL and can be done in as little as 10 minutes.

  6. Cryptosporidium infection in a veal calf cohort in France: molecular characterization of species in a longitudinal study

    Directory of Open Access Journals (Sweden)

    Follet Jérôme

    2011-12-01

    Full Text Available Abstract Feces from 142 animals were collected on 15 farms in the region of Brittany, France. Each sample was directly collected from the rectum of the animal and identified with the ear tag number. Animals were sampled three times, at 5, 15 and 22 weeks of age. After DNA extraction from stool samples, nested PCR was performed to amplify partial 18S-rDNA and 60 kDa glycoprotein genes of Cryptosporidium. The parasite was detected on all farms. One hundred out of 142 calves (70.4% were found to be parasitized by Cryptosporidium. Amplified fragments were sequenced for Cryptosporidium species identification and revealed the presence of C. parvum (43.8%, C. ryanae (28.5%, and C. bovis (27%. One animal was infected with Cryptosporidium ubiquitum. The prevalence of these species was related to the age of the animal. C. parvum caused 86.7% of Cryptosporidium infections in 5-week-old calves but only 1.7% in 15-week-old animals. The analysis of the results showed that animals could be infected successively by C. parvum, C. ryanae, and C. bovis for the study period. C. parvum gp60 genotyping identifies 6 IIa subtypes of which 74.5% were represented by IIaA15G2R1. This work confirms previous studies in other countries showing that zoonotic C. parvum is the dominant species seen in young calves.

  7. Occurrence of Cryptosporidium and Giardia in recycled waters used for irrigation and first description of Cryptosporidium parvum and C. muris in Greece.

    Science.gov (United States)

    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.

  8. Adhesion of Cryptosporidium parvum and Giardia lamblia to solid surfaces: the role of surface charge and hydrophobicity

    NARCIS (Netherlands)

    Dai, X.; Boll, J.; Hayes, M.E.; Aston, D.E.

    2004-01-01

    Adhesion of Cryptosporidium parvum and Giardia lamblia to four materials of different surface charge and hydrophobicity was investigated. Glass beads were used with and without three polymer coatings: aminosilines (A0750), fluorosilines (T2494), an amino cationic polymer. Surface charge density and

  9. Adhesion of Cryptosporidium parvum and Giardia lamblia to solid surfaces: the role of surface charge and hydrophobicity

    NARCIS (Netherlands)

    Dai, X.; Boll, J.; Hayes, M.E.; Aston, D.E.

    2004-01-01

    Adhesion of Cryptosporidium parvum and Giardia lamblia to four materials of different surface charge and hydrophobicity was investigated. Glass beads were used with and without three polymer coatings: aminosilines (A0750), fluorosilines (T2494), an amino cationic polymer. Surface charge density and

  10. Wastewater treatment with Moringa oleifera seed extract and impact on turbidity and sedimentation of Cryptosporidium parvum oocysts

    DEFF Research Database (Denmark)

    Petersen, Heidi Huus; Woolsey, Ian David; Dalsgaard, Anders

    produced from seeds of the Moringa oleifera tree (MO) in reducing Cryptosporidium parvum oocysts and turbidity in wastewater. To a total of 5 x 12 glass jars containing 500 ml wastewater samples from a Danish treatment plant, 1.2 x 106 ± 1.2 x 105 oocysts L-1 were added. To half of the wastewater samples 8...

  11. Stress-Induced Hsp70 Gene Expression and Inactivation of Cryptosporidium parvum Oocysts by Chlorine-Based Oxidants▿

    Science.gov (United States)

    Bajszár, George; Dekonenko, Alexander

    2010-01-01

    Our research on the mechanisms of action of chlorine-based oxidants on Cryptosporidium parvum oocysts in water revealed a dual-phase effect: (i) response to oxidative stress, which was demonstrated by induced expression of the Hsp70 heat shock gene, and (ii) oocyst inactivation as a result of long-term exposure to oxidants. The relative biocidal effects of sodium hypochlorite (bleach) and electrolytically generated mixed oxidant solution (MOS) on C. parvum oocysts were compared at identical free chlorine concentrations. Oocyst inactivation was determined by quantitative reverse transcription-PCR (qRT-PCR) amplification of the heat-induced Hsp70 mRNA and compared with tissue culture infectivity. According to both assays, within the range between 25 and 250 mg/liter free chlorine and with 4 h contact time, MOS exhibits a higher efficacy in oocyst inactivation than hypochlorite. Other RNA-based viability assays, aimed at monitoring the levels of β-tubulin mRNA and 18S rRNA, showed relatively slow decay rates of these molecules following disinfection by chlorine-based oxidants, rendering these molecular diagnostic viability markers inappropriate for disinfection efficacy assessment. PMID:20118357

  12. Triazole inhibitors of Cryptosporidium parvum inosine 5′-monophosphate dehydrogenase

    Science.gov (United States)

    Maurya, Sushil K.; Gollapalli, Deviprasad R.; Kirubakaran, Sivapriya; Zhang, Minjia; Johnson, Corey R.; Benjamin, Nicole N.; Hedstrom, Lizbeth; Cuny, Gregory D.

    2010-01-01

    Cryptosporidium parvum is an important human pathogen and potential bioterrorism agent. This protozoan parasite cannot salvage guanine or guanosine and therefore relies on inosine 5′-monophosphate dehydrogenase (IMPDH) for biosynthesis of guanine nucleotides and hence for survival. Since C. parvum IMPDH is highly divergent from the host counterpart, selective inhibitors could potentially be used to treat cryptosporidiosis with minimal effects on its mammalian host. A series of 1,2,3-triazole containing ether CpIMPDH inhibitors are described. A structure-activity relationship study revealed that a small alkyl group on the alpha-position of the ether was required with the (R)-enantiomer significantly more active than the (S)-enantiomer. Electron-withdrawing groups in the 3- and/or 4-positions of the pendent phenyl ring were best and conversion of the quinoline containing inhibitors to quinoline-N-oxides retained inhibitory activity both in the presence and absence of bovine serum albumin. The 1,2,3-triazole CpIMPDH inhibitors provide new tools for elucidating the role of IMPDH in C. parvum and may serve as potential therapeutics for treating cryptosporidiosis. PMID:19624136

  13. Pomegranate (Punica granatum) peel is effective in a murine model of experimental Cryptosporidium parvum ultrastructural studies of the ileum.

    Science.gov (United States)

    Al-Mathal, Ebtisam M; Alsalem, Afaf A

    2013-08-01

    The current treatments for cryptosporidiosis are ineffective, and there is an urgent need to search for more effective and safer alternatives. One such alternative may be treatments derived from natural resources. The pomegranate peel has been used effectively in traditional medicine to cure diarrhea and dysentery. The purpose of this study was to examine the effectiveness of a Punica granatum (pomegranate) peel suspension as a treatment for Cryptosporidium parvum infection. In this study, the effects of this treatment on the ultrastructure of both the intestinal epithelial layer of infected nursling mice and the parasite were observed with a transmission electron microscope. The histological study focused on the examination of the microvilli, columnar epithelium, goblet cells, lamina propria, and crypts of Lieberkuhn. Examination of the ileums of infected mice that received the pomegranate peel suspension demonstrated that the general structure of the ileal tissue of these mice was similar to that of the control group. In the infected mice treated with the suspension, but not the infected/untreated mice, there was an improvement in all ultrastructure aspects at 28days post-inoculation. The study of the ultrastructure of the parasite (C. parvum) in mice treated with the suspension showed that there was decomposition in the parasite to the extent that in some cases we were unable to identify the stage of the parasite due to the severe degeneration. Significant decomposition of the nutrition organ was also observed. Additionally, microgamonte and macrogamonte were not observed in the suspension-treated group, explaining the disappearance of the sexual phases of the parasite in the lumens of this group. In all, this examination demonstrated the restoration of the normal structures of villi and the disappearance of acute symptoms in the suspension-treated mice and showed that the suspension directly affected the parasite at various stages of its development and led to

  14. Movement of Cryptosporidium parvum Oocysts through Soils without Preferential Pathways: Exploratory Test

    Directory of Open Access Journals (Sweden)

    Christophe J. G. Darnault

    2017-06-01

    Full Text Available Groundwater contamination by oocysts of the waterborne pathogen Cryptosporidium parvum is a significant cause of animal and human disease worldwide. Although research has been undertaken in the past to determine how specific physical and chemical properties of soils affect the risk of groundwater contamination by C. parvum, there is as yet no clear conclusion concerning the range of mobility of C. parvum that one should expect in field soils. In this context, the key objective of this research was to determine the magnitude of C. parvum transport in a number of soils, under conditions in which fast and preferential transport has been successfully prevented. C. parvum oocysts were applied at the surface of different soils and subjected to artificial rainfall. Apparently for the first time, quantitative PCR was used to detect and enumerate oocysts in the soil columns and in the leachates. The transport of oocysts by infiltrating water, and the considerable retention of oocysts in soil was demonstrated for all soils, although differences in the degree of transport were observed with soils of different types. More oocysts were found in leachates from sandy loam soils than in leachates from loamy sand soils and the retention of oocysts in different soils did not significantly differ. The interaction of various processes of the hydrologic system and biogeochemical mechanisms contributed to the transport of oocysts through the soil matrix. Results suggest that the interplay of clay, organic matter, and Ca2+ facilitates and mediates the transfer of organic matter from mineral surfaces to oocysts surface, resulting in the enhanced breakthrough of oocysts through matrices of sandy loam soils compared to those of loamy sand soils. Although the number of occysts that penetrate the soil matrix account for only a small percentage of initial inputs, they still pose a significant threat to human health, especially in groundwater systems with a water table not

  15. [A new trichromic safranin stain for the detection of Cryptosporidium parvum, Cyclospora cayetanensis, species of Microsporidia and Isospora belli in fecal material].

    Science.gov (United States)

    Ponce de León, P; Flaherty, P; Zdero, M

    1999-01-01

    Cryptosporidium parvum, Isospora belli, Cyclospora cayetanensis and Microsporidia are frequent pathogens in the immunodeficient host, which may cause multiple infections. The above mentioned parasites are found in feces by the application of different specific tintorial techniques. The objective of this work was the development of a stain for the simultaneous detection of these parasites, reducing costs as well as the time taken to make the diagnosis. The safranin-trichrome stain is simple, chip and its results are similar to those of specific tints. All microorganisms are easy to detect and besides being perfectly distinguishable from fungi and faecal elements.

  16. Presence of Cryptosporidium scrofarum, C. suis and C. parvum subtypes IIaA16G2R1 and IIaA13G1R1 in Eurasian wild boars (Sus scrofa).

    Science.gov (United States)

    García-Presedo, Ignacio; Pedraza-Díaz, Susana; González-Warleta, Marta; Mezo, Mercedes; Gómez-Bautista, Mercedes; Ortega-Mora, Luis Miguel; Castro-Hermida, José Antonio

    2013-09-23

    The aim of the present study was to identify the species of Cryptosporidium infecting Eurasian wild boars (Sus scrofa) in Galicia (NW, Spain). A sampling of 209 wild boars shot in different game preserves was carried out during the hunting season in 2009-2010. All samples were examined for Cryptosporidium infection, using both immunological and molecular tools. Cryptosporidium oocysts in faecal samples were identified using a direct immunofluorescence technique with monoclonal antibodies (DFA). The presence of Cryptosporidium DNA was determined using nested PCR involving amplification of a fragment of the small-subunit (SSU) ribosomal RNA gene (SSU rRNA). A total of 35 (16.7%) samples tested positive with both techniques. However, sequencing was only possible in 27 samples. Cryptosporidium scrofarum, Cryptosporidium suis and Cryptosporidium parvum oocysts were identified in 19, 5 and 3 of the samples, respectively. Moreover, C. scrofarum was detected as a dominant species infecting all age groups (juveniles, sub adults and adults). Sequence analyses of the glycoprotein (GP60) gene revealed the presence of C. parvum subtypes IIaA16G2R1 in 2 juveniles and IIaA13G1R1 in 1 sub adult wild boar. These species and subtypes have previously been described in human patients, indicating that isolates from asymptomatic wild boars might have zoonotic potential. This is the first report of the presence of C. scrofarum, C. suis and C. parvum subtypes IIaA16G2R1 and IIaA13G1R1 in wild boars (S. scrofa) in Spain.

  17. Simultaneous detection of Giardia lamblia and Cryptosporidium parvum (oo)cysts in soil using immunomagnetic separation and direct fluorescent antibody staining.

    Science.gov (United States)

    Orlofsky, Ezra; Gillor, Osnat; Melli, Ann; Miller, Woutrina; Wuertz, Stefan; Bernstein, Nirit; Shapiro, Karen

    2013-09-01

    An improved approach for simultaneous detection of Cryptosporidium parvum and Giardia lamblia (oo)cysts in soil is described. Recoveries>70% were obtained for concentrations>55 and 21 (oo)cysts g(-1) for C. parvum and G. lamblia, respectively. The limits of detection were determined to be<5 (oo)cysts g(-1) soil.

  18. Risk of Handling as a Route of Exposure to Infectious Waterborne Cryptosporidium parvum Oocysts via Atlantic Blue Crabs (Callinectes sapidus)▿

    Science.gov (United States)

    Graczyk, Thaddeus K.; McOliver, Cynthia; Silbergeld, Ellen K.; Tamang, Leena; Roberts, Jennifer D.

    2007-01-01

    Commercial Atlantic blue crabs (Callinectes sapidus) were exposed to 2.0 × 104 infectious waterborne oocysts of Cryptosporidium parvum. The study demonstrated that blue crabs can transfer C. parvum oocysts to persons involved in handling or preparing crabs and that they may contaminate other surfaces or products during storage. PMID:17449680

  19. DETECTION OF PBMC T CELL SUBSETS AND mIL-2R OF THE PERSONS INFECTED BY CRYPTOSPORIDIUM PARVUM BY USING THE SYSTEM OF BSA%应用BSA系统检测隐孢子虫感染者外周血T细胞亚群及mIL-2R

    Institute of Scientific and Technical Information of China (English)

    许礼发; 李朝品; 张荣波; 王晓秋

    2004-01-01

    Objective To explore the changes of cellular immune function in the persons infected by Cryptosporidium parvum .MethodsThe system of biotin streptavidin (BSA) was used to detect T cell subsets and membrane interleukin 2 receptor (mIL 2R) in peripheral blood mononuclear cell (PBMC) of the persons infected byC. parvum . ResultsThe positive rates of CD3+,CD4+,CD8+ and CD4+/CD8+ in the persons in whose stoolsC. parvum oocysts were found were (54.77±7.33)%,(38.17±7.52)%,(30.64±5.87)% and 1.28±0.72 respectively,and the level of mIL 2R in induction period was (32.15±2.42)%. It could be concluded that the positive rates of CD3+,CD4+,CD8+ and CD4+/CD8+ in the persons in whose stoolsC. parvum oocysts were found were significantly different from those with noC. parvum oocysts were found (P <0.05~0.01). The level of mIL 2R in induction period was significantly different from which in resting period (P <0.05~0.01).ConclusionIn the persons infected byC. parvum,cellular immunity participates in resisting the parasite,and T cell subsets and mIL 2R play an important role.%目的探讨隐孢子虫感染者机体细胞免疫功能的变化. 方法采用生物素-链霉亲和素(Biotin-Streptavidin , BSA)法对卵囊阳性患者分别进行外周血T细胞亚群、膜白介素-2受体(mIL-2R)检测. 结果隐孢子虫卵囊阳性者的CD3+、CD4+、CD8+百分率和CD4+/CD8+比值分别为(54.77±7.33)%、(38.17±7.52)%、(30.64±5.87)% 和1.28±0.72,诱导期mIL-2R表达水平为(32.15±2.42)%,两者分别与卵囊阴性者及静息期mIL-2R相比,差异有显著性(P<0.05~0.01). 结论隐孢子虫感染者引起细胞免疫功能的变化,T细胞亚群、mIL-2R在抗隐孢子虫感染中起重要作用.

  20. Biological characterisation of Cryptosporidium parvum isolates of wildlife rodents in Poland.

    Science.gov (United States)

    Bednarska, Malgorzata; Bajer, Anna; Kulis, Karolina; Sinski, Edward

    2003-01-01

    The study was undertaken to characterise the C. parvum isolates originating from naturally infected woodland and field rodents: Clethrionomys glareolus (CG), Apodemus flavicollis (AF) and Microtus arvalis (MA). We found that the measurements of oocyst dimensions and oocyst morphology did not allow distinction between the parasite isolates from the 3 rodent species. The mean dimensions were: for CG 4.67 x 4.21, for AF 4.65 x 4.14 and for MA 4.66 x 4.16. These 3 groups of isolates have produced significantly different pictures of infection in C57BL/6 mice. The overall mean oocysts output was: in CG-mice 41,739, in AF-mice 18,000, in MA-mice 10,384 oocysts/1 g of faeces. From these data we suggest that rodent isolates of C. parvum could represent new subgroups in so-called "mouse" C. parvum strain. The successful cross-transmission from wild hosts to laboratory rodents and the close similarity of COWP sequence between our isolates and "mouse" genotype and between "mouse" and zoonotic genotype of C. parvum (genotype C) inform us that all these isolates should be treated as potentially hazardous for human health.

  1. Prevalencia del protozoario Cryptosporidium parvum en caninos en tres consultorios veterinarios en Tunja

    Directory of Open Access Journals (Sweden)

    Elkin Yadir Rodríguez-Becerra

    2011-02-01

    Full Text Available Se determinó por primera vez la prevalencia de Cryptosporidium parvum en caninos, en tres consultorios veterinarios de la ciudad de Tunja/Colombia, durante el periodo de tiempo comprendido entre agosto y diciembre de 2006. En el estudio se procesaron 132 muestras de materia fecal de perros, en el laboratorio clínico de la Universidad Pedagógica y Tecnológica de Colombia, UPTC, para lo cual se tuvo en cuenta su edad, raza y sexo, si presentaba diarrea, si se encontraba vacunado y/o desparasitado y, por último, a cada una de las muestras se le realizó la coloración de Ziehl – Nielsen modificada. El análisis estadístico se realizó mediante el programa Epiinfo. Del total de muestras examinadas, el 16,38% de los caninos fueron positivos a C. parvum. Del total de las muestras remitidas, un 47% correspondió a hembras y un 52,3% machos, con edades entre un mes y 13 años, con un promedio de tres años, de los cuales 34,8% presentaron diarrea. Las muestras positivas a C. parvum  se encontraron en perros entre 1 mes y 13 años de edad, con un promedio de cuatro años.

  2. Optimization of benzoxazole-based inhibitors of Cryptosporidium parvum inosine 5'-monophosphate dehydrogenase.

    Science.gov (United States)

    Gorla, Suresh Kumar; Kavitha, Mandapati; Zhang, Minjia; Chin, James En Wai; Liu, Xiaoping; Striepen, Boris; Makowska-Grzyska, Magdalena; Kim, Youngchang; Joachimiak, Andrzej; Hedstrom, Lizbeth; Cuny, Gregory D

    2013-05-23

    Cryptosporidium parvum is an enteric protozoan parasite that has emerged as a major cause of diarrhea, malnutrition, and gastroenteritis and poses a potential bioterrorism threat. C. parvum synthesizes guanine nucleotides from host adenosine in a streamlined pathway that relies on inosine 5'-monophosphate dehydrogenase (IMPDH). We have previously identified several parasite-selective C. parvum IMPDH (CpIMPDH) inhibitors by high-throughput screening. In this paper, we report the structure-activity relationship (SAR) for a series of benzoxazole derivatives with many compounds demonstrating CpIMPDH IC50 values in the nanomolar range and >500-fold selectivity over human IMPDH (hIMPDH). Unlike previously reported CpIMPDH inhibitors, these compounds are competitive inhibitors versus NAD(+). The SAR study reveals that pyridine and other small heteroaromatic substituents are required at the 2-position of the benzoxazole for potent inhibitory activity. In addition, several other SAR conclusions are highlighted with regard to the benzoxazole and the amide portion of the inhibitor, including preferred stereochemistry. An X-ray crystal structure of a representative E·IMP·inhibitor complex is also presented. Overall, the secondary amine derivative 15a demonstrated excellent CpIMPDH inhibitory activity (IC50 = 0.5 ± 0.1 nM) and moderate stability (t1/2 = 44 min) in mouse liver microsomes. Compound 73, the racemic version of 15a, also displayed superb antiparasitic activity in a Toxoplasma gondii strain that relies on CpIMPDH (EC50 = 20 ± 20 nM), and selectivity versus a wild-type T. gondii strain (200-fold). No toxicity was observed (LD50 > 50 μM) against a panel of four mammalian cells lines.

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

    Science.gov (United States)

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

    2014-12-15

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

  4. Transport and survival of Cryptosporidium Parvum Oocysts in Soil Columns Following Applications of Raw and Separated Liquid Slurry

    DEFF Research Database (Denmark)

    Petersen, H.H.; Enemark, Heidi L.; Olsen, A.;

    The widespread waterborne pathogen Cryptosporidium parvum is primarily transmitted to humans via contaminated drinking and recreational water. Nearly all drinking water in Denmark is groundwater, but this can be contaminated with oocysts from application of contaminated manure to the field. Oocysts...... transport to groundwater requires that the oocysts are transported through soil and bedrock to the water table. The purpose of this study was to determine the potential transport of the protozoan pathogen C. parvum through soil to land drains and, subsequently water courses in a laboratory setup using...... simulated rainfall and six 20 cm long replicate intact soil columns. Two types of contaminated slurry, namely raw slurry and the separated liquid fraction of the slurry were applied ten cm into the soil, following irrigation once a week over a four week period. C. parvum oocysts were detected...

  5. Cryptosporidium muris: Infectivity and Illness in Healthy Adult Volunteers

    Science.gov (United States)

    Chappell, Cynthia L.; Okhuysen, Pablo C.; Langer-Curry, Rebecca C.; Lupo, Philip J.; Widmer, Giovanni; Tzipori, Saul

    2015-01-01

    Although Cryptosporidium parvum and C. hominis cause the majority of human cryptosporidiosis cases, other Cryptosporidium species are also capable of infecting humans, particularly when individuals are immunocompromised. Ten C. muris cases have been reported, primarily in human immunodeficiency virus (HIV) -positive patients with diarrhea. However, asymptomatic cases were reported in two HIV-negative children, and in another case, age and immune status were not described. This study examines the infectivity of C. muris in six healthy adults. Volunteers were challenged with 105 C. muris oocysts and monitored for 6 weeks for infection and/or illness. All six patients became infected. Two patients experienced a self-limited diarrheal illness. Total oocysts shed during the study ranged from 6.7 × 106 to 4.1 × 108, and the number was slightly higher in volunteers with diarrhea (2.8 × 108) than asymptomatic shedders (4.4 × 107). C. muris-infected subjects shed oocysts longer than occurred with other species studied in healthy volunteers. Three volunteers shed oocysts for 7 months. Physical examinations were normal, with no reported recurrence of diarrhea or other gastrointestinal complaints. Two persistent shedders were treated with nitazoxanide, and the infection was resolved. Thus, healthy adults are susceptible to C. muris, which can cause mild diarrhea and result in persistent, asymptomatic infection. PMID:25311695

  6. Obtaining hyperimmune anti-Cryptosporidium parvum ovine colostrum. A study of the humoral immune response in immunized sheep.

    Science.gov (United States)

    Martín-Gómez, S; Alvarez-Sánchez, M A; Rojo-Vázquez, F A

    2006-01-01

    Three ewes were immunized five times over a 2-month period prior to giving birth by intramuscular injection, oral administration and intramammary infusion of antigen and viable or freeze-dried Cryptosporidium parvum oocyst solution emulsified with Freund's complete and incomplete adjuvant. Two animals served as controls and another two as adjuvant controls. Serum was collected at first immunization and thereafter every 2 to 4 weeks. Colostrum and milk were collected as well. All samples were assayed for C. parvum-specific antibodies using an enzyme-linked immunosorbent assay methodology, and Western blotting was used to recognize the C. parvum antigens. Hyperimmunization resulted in a progressive and significant increase in specific anti-C. parvum serum IgG, IgA and IgM titres, with the highest values noted at the point of lambing. Titres decreased slightly in milk, although they were in all cases higher than those in the control animals. Moreover, some 30 bands of C. parvum were recognized.

  7. THE EFFICACY OF THREE MEDICINAL PLANTS; GARLIC, GINGER AND MIRAZID AND A CHEMICAL DRUG METRONIDAZOLE AGAINST CRYPTOSPORIDIUM PARVUM: II-HISTOLOGICAL CHANGES.

    Science.gov (United States)

    Abouel-Nour, Mohamed F; El-Shewehy, Dina Magdy M; Hamada, Shadia F; Morsy, Tosson A

    2016-04-01

    Cryptosporidiosis parvum is a zoonotic protozoan parasite infects intestinal epithelial cells of man and animals causing a major health problem. This study was oriented to evaluate the protective and curative capacity of garlic, ginger and mirazid in comparison with metronidazole drug (commercially known) against Cryptosporidium in experimental mice. Male Swiss Albino mice experimentally infected with C. parvum were treated with medicinal plants extracts (Ginger, Mirazid, and Garlic) as compared to chemical drug Metronidazole. Importantly, C. parvum-infected mice treated with ginger, Mirazid, garlic and metronidazole showed a complete elimination in shedding oocysts by 9th day PI. The reduction and elimination of shedding oocysts in response to the treatments might be attributable to a direct effect on parasite growth in intestines, sexual phases production and/or the formation of oocysts. The results were evaluated histopathological examination of ideum section of control mice (uninfected, untreated) displayed normal architecture of the villi. Examiination of infected mice ileum section (infected, untreated) displayed histopathological alterations from uninfected groups. Examination of ileum section prepared from mice treated with garlic, ginger, mirazid, and metronidazole displayed histopathological alterations from that of the control groups, and showed marked histologic correction in the pattern with the four regimes used in comparison to control mice. Garlic successfully eradicated oocysts of infected mice from stool and intestine. Supplementation of ginger to infected mice markedly corrected elevation in the inflammatory risk factors and implied its potential antioxidant, anti-inflammatory and immunomodulatory capabilities. Infected mice treated with ginger, mirazid, garlic and metronidazole showed significant symptomatic improvements during treatment.

  8. Cryptosporidium Infections Among Children in Peru

    Centers for Disease Control (CDC) Podcasts

    2008-09-25

    Cryptosporidium is a waterborne bacteria that can cause severe diarrhea and vomiting. In this podcast, Dr. Vita Cama, CDC microbiologist, discusses an article in the October 2008 issue of Emerging Infectious Diseases. The paper examines Cryptosporidium infections among children in Peru, including the number of infections, symptoms experienced, and what species of Crypto were responsible.  Created: 9/25/2008 by Emerging Infectious Diseases.   Date Released: 9/25/2008.

  9. Expression of Cryptosporidium parvum thioredoxin peroxidase in COS-7 cells confers radioprotection.

    Science.gov (United States)

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

    2016-04-01

    Cryptosporidium parvum is one of the most radioresistant organisms identified to date. In a previous study, we found that thioredoxin peroxidase (CpTPx) was significantly upregulated in this species following exposure to high dose (10 kGy) of γ-irradiation. To assess the potential of CpTPx to confer radioprotection in mammalian cells, it was expressed in COS-7 African green monkey kidney cells (CpTPx-COS7). For comparison, the thioredoxin peroxidase of Cryptosporidium muris (CmTPx) was also expressed in these cells (CmTPx-COS7 cells), which has been confirmed to have lesser antioxidant activity than CpTPx in the previous study. Notably, the survival rates of CpTPx-COS7 cells were significantly higher (12-22%) at 72 h after 8 Gy irradiation than CmTPx-COS7 or non-transfected COS-7 (ntCOS-7) counterparts. In addition, CpTPx revealed a 50% of ROS reduction in irradiated CpTPx-COS7 cells, while γ-H2AX DNA damage marker expression was not significantly changed. Furthermore, the amount of apoptosis only increased to about 120% after 2-8 Gy irradiation compared to 200-300% increase observed in ntCOS-7 cells. CmTPx was shown to have antioxidant and DNA damage protection activities; however, these activities were always lower than those of CpTPx. These results suggest that the potent antioxidant and protective activities of CpTPx are well conserved in this cell-based system and that CpTPx contributed to the radioprotection of mammalian cells through its exceptional antioxidant activity.

  10. Multiplex-Touchdown PCR to Simultaneously Detect Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the Major Causes of Traveler’s Diarrhea

    Science.gov (United States)

    Shin, Ji-Hun; Lee, Sang-Eun; Kim, Tong Soo; Ma, Da-Won; Chai, Jong-Yil; Shin, Eun-Hee

    2016-01-01

    This study aimed to develop a multiplex-touchdown PCR method to simultaneously detect 3 species of protozoan parasites, i.e., Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the major causes of traveler’s diarrhea and are resistant to standard antimicrobial treatments. The target genes included the Cryptosporidium oocyst wall protein for C. parvum, Glutamate dehydrogenase for G. lamblia, and 18S ribosomal RNA (18S rRNA) for C. cayetanensis. The sizes of the amplified fragments were 555, 188, and 400 bps, respectively. The multiplex-touchdown PCR protocol using a primer mixture simultaneously detected protozoa in human stools, and the amplified gene was detected in >1×103 oocysts for C. parvum, >1×104 cysts for G. lamblia, and >1 copy of the 18S rRNA gene for C. cayetanensis. Taken together, our protocol convincingly demonstrated the ability to simultaneously detect C. parvum, G. lamblia, and C. cayetanenesis in stool samples. PMID:27853120

  11. Cryptosporidium species from human immunodeficiency-infected patients with chronic diarrhea in Jakarta, Indonesia.

    Science.gov (United States)

    Kurniawan, Agnes; Dwintasari, Sri W; Connelly, Lisa; Nichols, Rosely A B; Yunihastuti, Evy; Karyadi, Teguh; Djauzi, Samsuridjal

    2013-11-01

    Cryptosporidium is an opportunistic parasite that manifests as chronic and severe diarrhea in the immune-compromised subject. We investigated the species of Cryptosporidium to understand the epidemiology, mode of transmission, response to treatment, and prevention. Polymerase chain reaction/restriction fragment length polymorphism of the 18 S rRNA gene and sequencing were performed on 41 Cryptosporidium-positive stools from 36 patients with HIV AIDS, which comprised 36 pretreatment stools and 5 stools after treatment with Paromomycin. C. hominis, C. meleagridis, C. felis, and C. parvum were detected; 28 of 36 (77.7%) patients were infected with C. hominis and two (5.5%) patients with multiple species of Cryptosporidium. Treatment with Paromomycin resulted in different outcomes, perhaps because patients harbored other intestinal parasitic infections. Multiple infection with various Cryptosporidium species in the presence of other intestinal parasites occurs in patients with HIV AIDS suffering from chronic diarrhea who are severely immune-compromised. Common transmission of Cryptosporidium is anthroponotic. Copyright © 2013 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

    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...

  13. The Cryptosporidium parvum ApiAP2 gene family: insights into the evolution of apicomplexan AP2 regulatory systems.

    Science.gov (United States)

    Oberstaller, Jenna; Pumpalova, Yoanna; Schieler, Ariel; Llinás, Manuel; Kissinger, Jessica C

    2014-07-01

    We provide the first comprehensive analysis of any transcription factor family in Cryptosporidium, a basal-branching apicomplexan that is the second leading cause of infant diarrhea globally. AP2 domain-containing proteins have evolved to be the major regulatory family in the phylum to the exclusion of canonical regulators. We show that apicomplexan and perkinsid AP2 domains cluster distinctly from other chromalveolate AP2s. Protein-binding specificity assays of C. parvum AP2 domains combined with motif conservation upstream of co-regulated gene clusters allowed the construction of putative AP2 regulons across the in vitro life cycle. Orthologous Apicomplexan AP2 (ApiAP2) expression has been rearranged relative to the malaria parasite P. falciparum, suggesting ApiAP2 network rewiring during evolution. C. hominis orthologs of putative C. parvum ApiAP2 proteins and target genes show greater than average variation. C. parvum AP2 domains display reduced binding diversity relative to P. falciparum, with multiple domains binding the 5'-TGCAT-3', 5'-CACACA-3' and G-box motifs (5'-G[T/C]GGGG-3'). Many overrepresented motifs in C. parvum upstream regions are not AP2 binding motifs. We propose that C. parvum is less reliant on ApiAP2 regulators in part because it utilizes E2F/DP1 transcription factors. C. parvum may provide clues to the ancestral state of apicomplexan transcriptional regulation, pre-AP2 domination. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  14. Revisiting the reference genomes of human pathogenic Cryptosporidium species: reannotation of C. parvum Iowa and a new C. hominis reference

    Science.gov (United States)

    Isaza, Juan P.; Galván, Ana Luz; Polanco, Victor; Huang, Bernice; Matveyev, Andrey V.; Serrano, Myrna G.; Manque, Patricio; Buck, Gregory A.; Alzate, Juan F.

    2015-01-01

    Cryptosporidium parvum and C. hominis are the most relevant species of this genus for human health. Both cause a self-limiting diarrhea in immunocompetent individuals, but cause potentially life-threatening disease in the immunocompromised. Despite the importance of these pathogens, only one reference genome of each has been analyzed and published. These two reference genomes were sequenced using automated capillary sequencing; as of yet, no next generation sequencing technology has been applied to improve their assemblies and annotations. For C. hominis, the main challenge that prevents a larger number of genomes to be sequenced is its resistance to axenic culture. In the present study, we employed next generation technology to analyse the genomic DNA and RNA to generate a new reference genome sequence of a C. hominis strain isolated directly from human stool and a new genome annotation of the C. parvum Iowa reference genome. PMID:26549794

  15. Cryptosporidium infection in infancy as a cause of malnutrition

    DEFF Research Database (Denmark)

    Mølbak, Kare; Andersen, M; Aaby, Peter

    1997-01-01

    Cryptosporidium parvum causes persistent diarrhea in young children in developing countries. To determine the interaction between nutritional status and cryptosporidiosis, an open cohort of 1064 children younger than 3 y of age was followed for 1441 child-years by weekly diarrhea recall visits. A...

  16. Prevalence and genetic characterization of Cryptosporidium spp. and Cystoisospora belli in HIV-infected patients

    Directory of Open Access Journals (Sweden)

    Dnieber Chagas Assis

    2013-06-01

    Full Text Available Cryptosporidium spp. and Cystoisospora belli are monoxenic protozoa that have been recognized as the causative agents of chronic diarrhea in immunocompromised individuals, especially HIV-infected subjects. The objective of this study was to evaluate the frequency of these intestinal protozoa in HIV-positive patients in the Triângulo Mineiro region of Brazil and to correlate the presence of these infections with clinical, epidemiological and laboratory data of the patients. Oocysts were detected in stool samples of 10 (16.9% of the 59 patients studied, while Cryptosporidium spp. were present in 10.1% (6/59 and C. belli in 6.7% (4/59. The frequency of these parasites was higher among patients with diarrheic syndrome and CD4+ T lymphocyte counts < 200 cells/mm 3 , demonstrating the opportunistic characteristic of these infections. A significant association was observed between the lack of adherence to antiretroviral therapy and the presence of Cryptosporidium spp. and/or C. belli. Parasitism with Cryptosporidium spp. was more frequent in February and April, the months following the period of high rainfall. The same was not observed for C. belli. Genetic characterization of two isolates led to the identification of Cryptosporidium parvum, one of the main species associated with the zoonotic transmission of cryptosporidiosis.

  17. Prevalence and risk factors associated with Cryptosporidium spp. infection in young domestic livestock in India.

    Science.gov (United States)

    Maurya, Prem Sagar; Rakesh, Radhamma Lakshmipathy; Pradeep, Balaraju; Kumar, Saroj; Kundu, Krishnendu; Garg, Rajat; Ram, Hira; Kumar, Ashok; Banerjee, Partha Sarathi

    2013-04-01

    A total of 938 faecal samples (461 cattle calves, 264 buffalo calves, 55 lambs, 116 kids and 42 piglets) from different livestock farms and individual small holdings in six targeted states of India were collected and screened by modified Ziehl-Neelsen staining technique to determine the prevalence of Cryptosporidium spp. and its association with age, sex, season and faecal consistency in domesticated animals. Overall, 16.2 % of the animals were positive for Cryptosporidium infection with prevalence of 16.3, 24.2, 1.8, 3.5 and 19.1 % in cattle calves, buffalo calves, lambs, kids and piglets, respectively. The prevalence of infection was significantly higher (p0.05) was recorded in females than in males. Seasons had a significant effect (p0.05) in post-monsoon than in monsoon season. A high degree of association was noticed between Cryptosporidium infection and diarrhoea in ruminants screened during the present study. But, in case of pigs, the prevalence was higher in non-diarrhoeic than in diarrhoeic animals. Genotyping of Cryptosporidium spp. based on nested PCR amplification of partial 18S rRNA and its subsequent digestion with SspI, VspI and MboII restriction enzymes revealed prevalence of Cryptosporidium parvum in representative number of positive samples of cattle, buffalo and goats.

  18. Biotin- and Glycoprotein-Coated Microspheres as Surrogates for Studying Filtration Removal of Cryptosporidium parvum in a Granular Limestone Aquifer Medium.

    Science.gov (United States)

    Stevenson, M E; Blaschke, A P; Toze, S; Sidhu, J P S; Ahmed, W; van Driezum, I H; Sommer, R; Kirschner, A K T; Cervero-Aragó, S; Farnleitner, A H; Pang, L

    2015-07-01

    Members of the genus Cryptosporidium are waterborne protozoa of great health concern. Many studies have attempted to find appropriate surrogates for assessing Cryptosporidium filtration removal in porous media. In this study, we evaluated the filtration of Cryptosporidium parvum in granular limestone medium by the use of biotin- and glycoprotein-coated carboxylated polystyrene microspheres (CPMs) as surrogates. Column experiments were carried out with core material taken from a managed aquifer recharge site in Adelaide, Australia. For the experiments with injection of a single type of particle, we observed the total removal of the oocysts and glycoprotein-coated CPMs, a 4.6- to 6.3-log10 reduction of biotin-coated CPMs, and a 2.6-log10 reduction of unmodified CPMs. When two different types of particles were simultaneously injected, glycoprotein-coated CPMs showed a 5.3-log10 reduction, while the uncoated CPMs displayed a 3.7-log10 reduction, probably due to particle-particle interactions. Our results confirm that glycoprotein-coated CPMs are the most accurate surrogates for C. parvum; biotin-coated CPMs are slightly more conservative, while unmodified CPMs are markedly overly conservative for predicting C. parvum removal in granular limestone medium. The total removal of C. parvum observed in our study suggests that granular limestone medium is very effective for the filtration removal of C. parvum and could potentially be used for the pretreatment of drinking water and aquifer storage recovery of recycled water.

  19. DEVELOPMENT OF A CT EQUATION TAKING INTO CONSIDERATION THE EFFECT OF LOT VARIABILITY ON THE INACTIVATION OF CRYPTOSPORIDIUM PARVUM OOCYSTS WITH OZONE

    Science.gov (United States)

    Cryptosporidium parvum oocysts are prevalent in surface water and ground water under the influence of surface water, and are difficult to inactivate using free chlorine, the most common disinfectant currently used for treating drinking water. In contrast, it has been shown...

  20. Biotin- and glycoprotein-coated microspheres: potential surrogates for studying filtration of cryptosporidium parvum in porous media.

    Science.gov (United States)

    Pang, Liping; Nowostawska, Urszula; Weaver, Louise; Hoffman, Gabrielle; Karmacharya, Anjuman; Skinner, Alexandra; Karki, Naveena

    2012-11-06

    Cryptosporidium parvum is a waterborne pathogen, yet no suitable surrogate has been established for quantifying its filtration removal in porous media. Carboxyl polystyrene microspheres with size, density, and shape similar to C. parvum were coated with biotin (free and containing amine, NH(2)) and glycoprotein. These biomolecules have isoelectric points similar to C. parvum (pH ≈ 2), and glycoprotein is a major type of surface protein that oocysts possess. Zeta potential (ζ) and filtration removal of particles in sand of two different grain sizes were examined. Compared to unmodified microspheres, modified microspheres achieved a superior match to the oocysts in ζ, concentration, mass recovery, and collision coefficient. They showed the same log reduction in concentration as oocysts, whereas results from unmodified microspheres deviated by 1 order of magnitude. Of the three types of modified microspheres, glycoprotein-coated microspheres best resembled oocyst concentration, despite having ζ similar to NH(2)-biotin-coated microspheres, suggesting that surface protein also played an important role in particle attachment on solid surfaces. With further validation in environmental conditions, the surrogates developed here could be a cost-effective new tool for assessing oocyst filtration in porous media, for example, to evaluate the performance of sand filters in water and wastewater treatment, water recycling through riverbank filtration, and aquifer recharge.

  1. Capture of water-borne colloids in granular beds using external electric fields: improving removal of Cryptosporidium parvum.

    Science.gov (United States)

    Kulkarni, Pramod; Dutari, Gabriel; Weingeist, David; Adin, Avner; Haught, Roy; Biswas, Pratim

    2005-03-01

    Suboptimal coagulation in water treatment plants often results in reduced removal efficiency of Cryptosporidium parvum oocysts by several orders of magnitude (J. AWWA 94(6) (2002) 97, J. AWWA 93(12) (2001) 64). The effect of external electric field on removal of C. parvum oocysts in packed granular beds was studied experimentally. A cylindrical configuration of electrodes, with granular media in the annular space was used. A negative DC potential was applied to the central electrode. No coagulants or flocculants were used and filtration was performed with and without application of an electric field to obtain improvement in removal efficiency. Results indicate that removal of C. parvum increased from 10% to 70% due to application of field in fine sand media and from 30% to 96% in MAGCHEM media. All other test particles (Kaolin and polystyrene latex microspheres) used in the study also exhibited increased removal in the presence of an electric field. Single collector efficiencies were also computed using approximate trajectory analysis, modified to account for the applied external electric field. The results of these calculations were used to qualitatively explain the trends in the experimental observations.

  2. Use of a sentinel system for field measurements of Cryptosporidium parvum oocyst inactivation in soil and animal waste.

    Science.gov (United States)

    Jenkins, M B; Walker, M J; Bowman, D D; Anthony, L C; Ghiorse, W C

    1999-05-01

    A small-volume sentinel chamber was developed to assess the effects of environmental stresses on survival of sucrose-Percoll-purified Cryptosporidium parvum oocysts in soil and animal wastes. Chambers were tested for their ability to equilibrate with external chemical and moisture conditions. Sentinel oocysts were then exposed to stresses of the external environment that affected their viability (potential infectivity), as indicated by results of a dye permeability assay. Preliminary laboratory experiments indicated that temperatures between 35 and 50 degrees C and decreases in soil water potential (-0.003 to -3.20 MPa) increased oocyst inactivation rates. The effects of two common animal waste management practices on oocyst survival were investigated on three dairy farms in Delaware County, N.Y., within the New York City watershed: (i) piling wastes from dairy youngstock (including neonatal calves) and (ii) spreading wastes as a soil amendment on an agricultural field. Sentinel containers filled with air-dried and sieved (2-mm mesh) youngstock waste or field soil were wetted and inoculated with 2 million oocysts in an aqueous suspension and then placed in waste piles on two different farms and in soil within a cropped field on one farm. Controls consisted of purified oocysts in either phosphate-buffered saline or distilled water contained in sealed microcentrifuge tubes. Two microdata loggers recorded the ambient temperature at each field site. Sentinel experiments were conducted during the fall and winter (1996 to 1997) and winter (1998). Sentinel containers and controls were removed at 2- to 4-week intervals, and oocysts were extracted and tested by the dye permeability assay. The proportions of potentially infective oocysts exposed to the soil and waste pile material decreased more rapidly than their counterpart controls exposed to buffer or water, indicating that factors other than temperature affected oocyst inactivation in the waste piles and soil. The

  3. Ureaplasma parvum infection alters filamin a dynamics in host cells

    Directory of Open Access Journals (Sweden)

    Brown Mary B

    2011-04-01

    Full Text Available Abstract Background Ureaplasmas are among the most common bacteria isolated from the human urogenital tract. Ureaplasmas can produce asymptomatic infections or disease characterized by an exaggerated inflammatory response. Most investigations have focused on elucidating the pathogenic potential of Ureaplasma species, but little attention has been paid to understanding the mechanisms by which these organisms are capable of establishing asymptomatic infection. Methods We employed differential proteome profiling of bladder tissues from rats experimentally infected with U. parvum in order to identify host cell processes perturbed by colonization with the microbe. Tissues were grouped into four categories: sham inoculated controls, animals that spontaneously cleared infection, asymptomatic urinary tract infection (UTI, and complicated UTI. One protein that was perturbed by infection (filamin A was used to further elucidate the mechanism of U. parvum-induced disruption in human benign prostate cells (BPH-1. BPH-1 cells were evaluated by confocal microscopy, immunoblotting and ELISA. Results Bladder tissue from animals actively colonized with U. parvum displayed significant alterations in actin binding proteins (profilin 1, vinculin, α actinin, and filamin A that regulate both actin polymerization and cell cytoskeletal function pertaining to focal adhesion formation and signal transduction (Fisher's exact test, P U. parvum perturbed the regulation of filamin A. Specifically, infected BPH-1 cells exhibited a significant increase in filamin A phosphorylated at serine2152 (P ≤ 0.01, which correlated with impaired proteolysis of the protein and its normal intracellular distribution. Conclusion Filamin A dynamics were perturbed in both models of infection. Phosphorylation of filamin A occurs in response to various cell signaling cascades that regulate cell motility, differentiation, apoptosis and inflammation. Thus, this phenomenon may be a useful

  4. Evaluation of solar photocatalysis using TiO2 slurry in the inactivation of Cryptosporidium parvum oocysts in water.

    Science.gov (United States)

    Abeledo-Lameiro, María Jesús; Ares-Mazás, Elvira; Gómez-Couso, Hipólito

    2016-10-01

    Cryptosporidium is a genus of enteric protozoan parasites of medical and veterinary importance, whose oocysts have been reported to occur in different types of water worldwide, offering a great resistant to the water treatment processes. Heterogeneous solar photocatalysis using titanium dioxide (TiO2) slurry was evaluated on inactivation of Cryptosporidium parvum oocysts in water. Suspensions of TiO2 (0, 63, 100 and 200mg/L) in distilled water (DW) or simulated municipal wastewater treatment plant (MWTP) effluent spiked with C. parvum oocysts were exposed to simulated solar radiation. The use of TiO2 slurry at concentrations of 100 and 200mg/L in DW yielded a high level of oocyst inactivation after 5h of exposure (4.16±2.35% and 15.03±4.54%, respectively, vs 99.33±0.58%, initial value), representing a good improvement relative to the results obtained in the samples exposed without TiO2 (51.06±9.35%). However, in the assays carried out using simulated MWTP effluent, addition of the photocatalyst did not offer better results. Examination of the samples under bright field and epifluorescence microscopy revealed the existence of aggregates comprising TiO2 particles and parasitic forms, which size increased as the concentration of catalyst and the exposure time increased, while the intensity of fluorescence of the oocyst walls decreased. After photocatalytic disinfection process, the recovery of TiO2 slurry by sedimentation provided a substantial reduction in the parasitic load in treated water samples (57.81±1.10% and 82.10±2.64% for 200mg/L of TiO2 in DW and in simulated MWTP effluent, respectively). Although further studies are need to optimize TiO2 photocatalytic disinfection against Cryptosporidium, the results obtained in the present study show the effectiveness of solar photocatalysis using TiO2 slurry in the inactivation of C. parvum oocysts in distilled water.

  5. Presence of Cryptosporidium parvum and Giardia lamblia in water samples from Southeast Asia: towards an integrated water detection system.

    Science.gov (United States)

    Kumar, Thulasi; Abd Majid, Mohamad Azlan; Onichandran, Subashini; Jaturas, Narong; Andiappan, Hemah; Salibay, Cristina C; Tabo, Hazel A L; Tabo, Norbel; Dungca, Julieta Z; Tangpong, Jitbanjong; Phiriyasamith, Sucheep; Yuttayong, Boonyaorn; Polseela, Raxsina; Do, Binh Nhu; Sawangjaroen, Nongyao; Tan, Tian-Chye; Lim, Yvonne A L; Nissapatorn, Veeranoot

    2016-01-13

    Access to clean and safe drinking water that is free from pathogenic protozoan parasites, especially Cryptosporidium parvum and Giardia lamblia that cause gastrointestinal illness in humans, is still an issue in Southeast Asia (SEA). This study is the first attempt to detect the aforementioned protozoan parasites in water samples from countries in SEA, using real-time polymerase chain reaction (qPCR) assays. A total of 221 water samples of 10 l each were collected between April and October 2013 from Malaysia (53), Thailand (120), the Philippines (33), and Vietnam (15). A physicochemical analysis was conducted. The water samples were processed in accordance with the US Environmental Protection Agency's methods 1622/1623.1, microscopically observed and subsequently screened using qPCR assays. Cryptosporidium oocysts were detected in treated water samples from the Philippines (1/10), with a concentration of 0.06 ± 0.19 oocyst/L, and untreated water samples from Thailand (25/93), Malaysia (17/44), and the Philippines (11/23), with concentrations ranging from 0.13 ± 0.18 to 0.57 ± 1.41 oocyst/L. Giardia cysts were found in treated water samples from the Philippines (1/10), with a concentration of 0.02 ± 0.06 cyst/L, and in untreated water samples from Thailand (20/93), Vietnam (5/10), Malaysia (22/44), and the Philippines (16/23), with concentrations ranging from 0.12 ± 0.3 to 8.90 ± 19.65 cyst/L. The pathogens C. parvum and G. lamblia were detected using using qPCR assays by targeting the 138-bp fragment and the small subunit gene, respectively. C. parvum was detected in untreated water samples from the Philippines (1/23) and Malaysia (2/44), whilst, G. lamblia detected was detected in treated water samples from the Philippines (1/10) and in untreated water samples from Thailand (21/93), Malaysia (12/44), and the Philippines (17/23). Nitrate concentration was found to have a high positive correlation with (oo)cyst (0.993). The presence of

  6. Prevalência do Cryptosporidium parvum em crianças abaixo de 5 anos, residentes na zona urbana de Campo Grande, MS, Brasil, 1996

    OpenAIRE

    Oshiro Elisa Teruya; Dorval Maria Elizabeth Cavalheiros; Nunes Vânia Lúcia Brandão; Silva Marcos Aurélio Almeida; Said Luis Augusto Morelli

    2000-01-01

    O presente estudo visou estabelecer a prevalência de Cryptosporidium parvum em crianças abaixo de 5 anos, residentes na zona urbana de Campo Grande, MS, 1996/97, através de exames coprológicos e avaliar epidemiologicamente os casos diagnosticados. Tratou-se de um estudo transversal com inquérito domiciliar, onde foram examinadas 1051 amostras fecais, processadas segundo a técnica de Blagg, utilizando-se a coloração de Ziehl-Neelsen modificada para a pesquisa de oocistos de C. parvum. Concluiu...

  7. Removal and fate of Cryptosporidium parvum, Clostridium perfringens and small-sized centric diatoms (Stephanodiscus hantzschii) in slow sand filters.

    Science.gov (United States)

    Hijnen, Wim A M; Dullemont, Yolanda J; Schijven, Jack F; Hanzens-Brouwer, Anke J; Rosielle, Martine; Medema, Gertjan

    2007-05-01

    The decimal elimination capacity (DEC) of slow sand filtration (SSF) for Cryptosporidium parvum was assessed to enable quantitative microbial risk analysis of a drinking water production plant. A mature pilot plant filter of 2.56m(2) was loaded with C. parvum oocysts and two other persistent organisms as potential surrogates; spores of Clostridium perfringens (SCP) and the small-sized (4-7microm) centric diatom (SSCD) Stephanodiscus hantzschii. Highly persistent micro-organisms that are retained in slow sand filters are expected to accumulate and eventually break through the filter bed. To investigate this phenomenon, a dosing period of 100 days was applied with an extended filtrate monitoring period of 150 days using large-volume sampling. Based on the breakthrough curves the DEC of the filter bed for oocysts was high and calculated to be 4.7log. During the extended filtrate monitoring period the spatial distribution of the retained organisms in the filter bed was determined. These data showed little risk of accumulation of oocysts in mature filters most likely due to predation by zooplankton. The DEC for the two surrogates, SCP and SSCD, was 3.6 and 1.8log, respectively. On basis of differences in transport behaviour, but mainly because of the high persistence compared to the persistence of oocysts, it was concluded that both spores of sulphite-reducing clostridia (incl. SCP) and SSCD are unsuited for use as surrogates for oocyst removal by slow sand filters. Further research is necessary to elucidate the role of predation in Cryptosporidium removal and the fate of consumed oocysts.

  8. Hydrologic and vegetative removal of Cryptosporidium parvum, Giardia lamblia, and Toxoplasma gondii Surrogate microspheres in coastal wetlands.

    Science.gov (United States)

    Hogan, Jennifer N; Daniels, Miles E; Watson, Fred G; Oates, Stori C; Miller, Melissa A; Conrad, Patricia A; Shapiro, Karen; Hardin, Dane; Dominik, Clare; Melli, Ann; Jessup, David A; Miller, Woutrina A

    2013-03-01

    Constructed wetland systems are used to reduce pollutants and pathogens in wastewater effluent, but comparatively little is known about pathogen transport through natural wetland habitats. Fecal protozoans, including Cryptosporidium parvum, Giardia lamblia, and Toxoplasma gondii, are waterborne pathogens of humans and animals, which are carried by surface waters from land-based sources into coastal waters. This study evaluated key factors of coastal wetlands for the reduction of protozoal parasites in surface waters using settling column and recirculating mesocosm tank experiments. Settling column experiments evaluated the effects of salinity, temperature, and water type ("pure" versus "environmental") on the vertical settling velocities of C. parvum, G. lamblia, and T. gondii surrogates, with salinity and water type found to significantly affect settling of the parasites. The mesocosm tank experiments evaluated the effects of salinity, flow rate, and vegetation parameters on parasite and surrogate counts, with increased salinity and the presence of vegetation found to be significant factors for removal of parasites in a unidirectional transport wetland system. Overall, this study highlights the importance of water type, salinity, and vegetation parameters for pathogen transport within wetland systems, with implications for wetland management, restoration efforts, and coastal water quality.

  9. Serum and colostrum antibody responses induced by jet-injection of sheep with DNA encoding a Cryptosporidium parvum antigen.

    Science.gov (United States)

    Jenkins, M; Kerr, D; Fayer, R; Wall, R

    1995-12-01

    In an effort to generate high titer colostrum for immunotherapy of cryptosporidiosis, a study was conducted to test the efficacy of immunizing sheep with recombinant plasmid DNA (pCMV-CP15/60) encoding epitopes of 15 and 60 kDa surface antigens of Cryptosporidium parvum sporozoites. The plasmid DNA was used to immunize preparturient ewes at three dose levels by jet-injection into either hind limb muscle (IM) or mammary tissue (IMAM). Regardless of route of injection, a dose-dependent anti-CP15/60 immunoglobulin response was observed in sera and colostrum from sheep immunized with pCMV-CP15/60 plasmid DNA. High titer antibody responses were observed in one of three animals per group receiving an IM injection of 100 or 1000 micrograms pCMV-CP15/60. IMAM immunization with 100 or 1000 micrograms pCMV-CP15/60 plasmid DNA elicited higher titer colostrum responses and more consistent serum responses compared to IM injections. A negligible serum and colostrum anti-CP15/60 response was observed in ewes injected IM with 10 micrograms pCMV-CP15/60 or 1000 micrograms control plasmid DNA. Immunoblotting of native C. parvum sporozoite/oocyst protein with hyperimmune serum and colostrum corroborated the increased titers against CP15/60 antigen. Serum and colostrum antibodies from pCMV-CP15/60-immunized sheep were eluted from native CP15 protein and bound a surface antigen of C. parvum sporozoites as indicated by indirect immunofluorescence staining.

  10. Comparative diagnostic techniques for cryptosporidium infection.

    Science.gov (United States)

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

    2014-02-24

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

  11. Comparative Diagnostic Techniques for Cryptosporidium Infection

    Directory of Open Access Journals (Sweden)

    Beauty E. Omoruyi

    2014-02-01

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

  12. Clinical and subclinical infections with Giardia and Cryptosporidium in animals

    Science.gov (United States)

    Giardia and Cryptosporidium are frequent parasites of livestock, companion animals, and wildlife, raising questions about the clinical significance of such infections. Infections with both parasites have a wide spectrum of symptoms that can vary between asymptomatic infections to serious infection ...

  13. Transcriptome analysis reveals unique metabolic features in the Cryptosporidium parvum Oocysts associated with environmental survival and stresses

    Directory of Open Access Journals (Sweden)

    Zhang Haili

    2012-11-01

    Full Text Available Abstract Background Cryptosporidium parvum is a globally distributed zoonotic parasite and an important opportunistic pathogen in immunocompromised patients. Little is known on the metabolic dynamics of the parasite, and study is hampered by the lack of molecular and genetic tools. Here we report the development of the first Agilent microarray for C. parvum (CpArray15K that covers all predicted ORFs in the parasite genome. Global transcriptome analysis using CpArray15K coupled with real-time qRT-PCR uncovered a number of unique metabolic features in oocysts, the infectious and environmental stage of the parasite. Results Oocyst stage parasites were found to be highly active in protein synthesis, based on the high transcript levels of genes associated with ribosome biogenesis, transcription and translation. The proteasome and ubiquitin associated components were also highly active, implying that oocysts might employ protein degradation pathways to recycle amino acids in order to overcome the inability to synthesize amino acids de novo. Energy metabolism in oocysts was featured by the highest level of expression of lactate dehydrogenase (LDH gene. We also studied parasite responses to UV-irradiation, and observed complex and dynamic regulations of gene expression. Notable changes included increased transcript levels of genes involved in DNA repair and intracellular trafficking. Among the stress-related genes, TCP-1 family members and some thioredoxin-associated genes appear to play more important roles in the recovery of UV-induced damages in the oocysts. Our observations also suggest that UV irradiation of oocysts results in increased activities in cytoskeletal rearrangement and intracellular membrane trafficking. Conclusions CpArray15K is the first microarray chip developed for C. parvum, which provides the Cryptosporidium research community a needed tool to study the parasite transcriptome and functional genomics. CpArray15K has been

  14. Inhibitory Activities of Epidermal Growth Factor Receptor Tyrosine Kinase-Targeted Dihydroxyisoflavone and Trihydroxydeoxybenzoin Derivatives on Sarcocystis neurona, Neospora caninum, and Cryptosporidium parvum Development

    Science.gov (United States)

    Gargala, G.; Baishanbo, A.; Favennec, L.; François, A.; Ballet, J. J.; Rossignol, J.-F.

    2005-01-01

    Several gene sequences of parasitic protozoa belonging to protein kinase gene families and epidermal growth factor (EGF)-like peptides, which act via binding to receptor tyrosine kinases of the EGF receptor (EGFR) family, appear to mediate host-protozoan interactions. As a clue to EGFR protein tyrosine kinase (PTK) mediation and a novel approach for identifying anticoccidial agents, activities against Sarcocystis neurona, Neospora caninum, and Cryptosporidium parvum grown in BM and HCT-8 cell cultures of 52 EGFR PTK inhibitor isoflavone analogs (dihydroxyisoflavone and trihydroxydeoxybenzoine derivatives) were investigated. Their cytotoxicities against host cells were either absent, mild, or moderate by a nitroblue tetrazolium test. At concentrations ranging from 5 to 10 μg/ml, 20 and 5 analogs, including RM-6427 and RM-6428, exhibited an in vitro inhibitory effect of ≥95% against at least one parasite or against all three, respectively. In immunosuppressed Cryptosporidium parvum-infected Mongolian gerbils orally treated with either 200 or 400 mg of agent RM-6427/kg of body weight/day for 8 days, fecal microscopic oocyst shedding was abolished in 6/10 animals (P of 0.05, respectively). After RM-6427 therapy (200 mg/kg/day for 8 days), the reduction in the ratio of animals with intracellular parasites was nearly significant in ileum (P = 0.067) and more marked in the biliary tract (P < 0.0013) than after nitazoxanide or paromomycin treatment (0.05 < P < 0.004). RM-6428 treatment at a regimen of 400 mg/kg/day for 12 days inhibited oocyst shedding, measured using flow cytometry from day 4 (P < 0.05) to day 12 (P < 0.02) of therapy, when 2/15 animals had no shedding (P < 0.0001) and 11/15 were free of gut and/or biliary tract parasites (P < 0.01). No mucosal alteration was microscopically observed for treated or untreated infected gerbils. To our knowledge, this report is the first to suggest that the isoflavone class of agents has the potential for

  15. Wastewater treatment with Moringa oleifera seed extract: Impact on turbidity and sedimentation of Cryptosporidium parvum oocysts

    OpenAIRE

    Petersen, Heidi H.; Woolsey, Ian; Dalsgaard, Anders; Enemark, Heidi; Olsen, Annette

    2014-01-01

    The use of low quality water for irrigation in agriculture is common practise in many countries due to limited freshwater resources. Pathogens may contaminate vegetables when faeces polluted water is used for irrigation. A laboratory study was carried out to investigate the effect of a coagulant produced from seeds of the Moringa oleifera tree (MO) in reducing Cryptosporidium parvum oocysts and turbidity in wastewater. To a total of 5 x 12 glass jars containing 500 ml wastewater samples from ...

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

    Science.gov (United States)

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

    2013-10-18

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

  17. Transport and survival of Cryptosporidium parvum oocysts in soil columns following applications of raw and separated liquid slurry

    DEFF Research Database (Denmark)

    Petersen, Heidi H.; Enemark, Heidi; Olsen, Annette;

    The widespread waterborne pathogen Cryptosporidium parvum is frequently transmitted to humans via contaminated drinking and recreational water. Nearly all drinking water in Denmark is groundwater, which can be contaminated with oocysts e.g. from application of contaminated manure to the field....... Oocysts transport to groundwater requires that the oocysts are transported through soil and bedrock to the water table. The purpose of this study was to determine the potential transport of the protozoan pathogen C. parvum through soil to land drains and, subsequently water sources in a laboratory setup...... using simulated rainfall and six 20 cm long replicate intact soil columns. Two types of contaminated slurry: raw slurry and the separated liquid fraction of the slurry were applied ten cm into the soil, which was subsequently irrigated once a week over a four-week period. C. parvum oocysts were detected...

  18. Evaluation of the BD MAX Enteric Parasite Panel for the detection of Cryptosporidium parvum/hominis, Giardia duodenalis and Entamoeba histolytica.

    Science.gov (United States)

    Perry, Michael D; Corden, Sally A; Lewis White, P

    2017-08-01

    Conventional laboratory detection methods for gastrointestinal parasites are time consuming, require considerable technical expertise and may suffer from poor analytical sensitivity. This study sought to evaluate the automated BD MAX Enteric Parasite Panel (EPP) for the detection of Cryptosporidium parvum/hominis, Entamoeba histolytica and Giardia duodenalis.Methodolgy. A total of 104 known positive samples (43 Cryptosporidium parvum/hominis and 61 G. duodenalis), 15 simulated samples (E. histolytica and other Entamoeba species) and 745 patient stool samples, submitted for enteric pathogen culture and microscopy, were inoculated into BD MAX EPP sample buffer tubes (SBTs). All specimens were blinded and tested within 7 days of SBT inoculation using the BD MAX EPP assay with results compared to those generated by microscopy.Results/Key findings. Combining the results from the known positive samples and anonymously tested patient samples, the sensitivity of the BD MAX EPP assay was 100 % for both Cryptosporidium spp. and G. duodenalis. Specificities of 99.7 and 98.9 % were calculated for the detection of Cryptosporidium spp. and G. duodenalis respectively. Insufficient clinical specimen data was available to determine the performance of the assay for E. histolytica detection. The findings of this study indicate that the BD MAX EPP is suitable for the detection of Cryptosporidium parvum/hominis and G. duodenalis from clinical specimens with reduced hands-on time and complexity compared to microscopy. Results for the detection of E. histolytica were promising although further work is required to evaluate the assay for the detection of this pathogen.

  19. Cryptosporidium parvum vaccine candidates are incompletely modified with O-linked-N-acetylgalactosamine or contain N-terminal N-myristate and S-palmitate.

    Science.gov (United States)

    Haserick, John R; Klein, Joshua A; Costello, Catherine E; Samuelson, John

    2017-01-01

    Cryptosporidium parvum (studied here) and Cryptosporidium hominis are important causes of diarrhea in infants and immunosuppressed persons. C. parvum vaccine candidates, which are on the surface of sporozoites, include glycoproteins with Ser- and Thr-rich domains (Gp15, Gp40, and Gp900) and a low complexity, acidic protein (Cp23). Here we used mass spectrometry to determine that O-linked GalNAc is present in dense arrays on a glycopeptide with consecutive Ser derived from Gp40 and on glycopeptides with consecutive Thr derived from Gp20, a novel C. parvum glycoprotein with a formula weight of ~20 kDa. In contrast, the occupied Ser or Thr residues in glycopeptides from Gp15 and Gp900 are isolated from one another. Gly at the N-terminus of Cp23 is N-myristoylated, while Cys, the second amino acid, is S-palmitoylated. In summary, C. parvum O-GalNAc transferases, which are homologs of host enzymes, densely modify arrays of Ser or Thr, as well as isolated Ser and Thr residues on C. parvum vaccine candidates. The N-terminus of an immunodominant antigen has lipid modifications similar to those of host cells and other apicomplexan parasites. Mass spectrometric demonstration here of glycopeptides with O-glycans complements previous identification C. parvum O-GalNAc transferases, lectin binding to vaccine candidates, and human and mouse antibodies binding to glycopeptides. The significance of these post-translational modifications is discussed with regards to the function of these proteins and the design of serological tests and vaccines.

  20. Removals of cryptosporidium parvum oocysts and cryptosporidium-sized polystyrene microspheres from swimming pool water by diatomaceous earth filtration and perlite-sand filtration.

    Science.gov (United States)

    Lu, Ping; Amburgey, James E; Hill, Vincent R; Murphy, Jennifer L; Schneeberger, Chandra L; Arrowood, Michael J; Yuan, Tao

    2017-06-01

    Removal of Cryptosporidium-sized microspheres and Cryptosporidium parvum oocysts from swimming pools was investigated using diatomaceous earth (DE) precoat filtration and perlite-sand filtration. In pilot-scale experiments, microsphere removals of up to 2 log were obtained with 0.7 kg·DE/m(2) at a filtration rate of 5 m/h. A slightly higher microsphere removal (2.3 log) was obtained for these DE-precoated filters when the filtration rate was 3.6 m/h. Additionally, pilot-scale perlite-sand filters achieved greater than 2 log removal when at least 0.37 kg/m(2) of perlite was used compared to 0.1-0.4 log removal without perlite both at a surface loading rate of 37 m/h. Full-scale testing achieved 2.7 log of microspheres and oocysts removal when 0.7 kg·DE/m(2) was used at 3.6 m/h. Removals were significantly decreased by a 15-minute interruption of the flow (without any mechanical agitation) to the DE filter in pilot-scale studies, which was not observed in full-scale filters. Microsphere removals were 2.7 log by perlite-sand filtration in a full-scale swimming pool filter operated at 34 m/h with 0.5 kg/m(2) of perlite. The results demonstrate that either a DE precoat filter or a perlite-sand filter can improve the efficiency of removal of microspheres and oocysts from swimming pools over a standard sand filter under the conditions studied.

  1. Prevalência do Cryptosporidium parvum em crianças abaixo de 5 anos, residentes na zona urbana de Campo Grande, MS, Brasil, 1996

    Directory of Open Access Journals (Sweden)

    Oshiro Elisa Teruya

    2000-01-01

    Full Text Available O presente estudo visou estabelecer a prevalência de Cryptosporidium parvum em crianças abaixo de 5 anos, residentes na zona urbana de Campo Grande, MS, 1996/97, através de exames coprológicos e avaliar epidemiologicamente os casos diagnosticados. Tratou-se de um estudo transversal com inquérito domiciliar, onde foram examinadas 1051 amostras fecais, processadas segundo a técnica de Blagg, utilizando-se a coloração de Ziehl-Neelsen modificada para a pesquisa de oocistos de C. parvum. Concluiu-se que: a prevalência de C. parvum (1,1% observada não foi estatisticamente significativa; foi relatado diarréia em 58,3% das crianças com diagnóstico positivo, supondo-se associação entre diarréia e a presença do parasita; o C. parvum foi mais freqüente em crianças com idade de 25 a 36 meses (50%, porém sem diferença estatisticamente significativa; o sexo não teve papel diferencial em relação ao parasitismo por C. parvum; entre as 12 crianças com criptosporidiose, 83,3% tiveram contato com animais domésticos (cão e ou gato.

  2. Construction of genomic libraries of Cryptosporidium parvum and identification of antigen-encoding genes.

    Science.gov (United States)

    Dykstra, C C; Blagburn, B L; Tidwell, R R

    1991-01-01

    Genomic libraries have been constructed from bovine C. parvum DNA in the lambda ZAP and lambda DASH vectors. Based on an estimated genome size of 2 x 10(4) kilobases (kb), each recombinant library contains greater than 10 genomic equivalents. The average recombinant size for the lambda ZAP library is 2.1 kb and for the lambda DASH library is 14 kb. We have identified genes to major antigens recognized by hyperimmune bovine antiserum. These recombinants are currently being purified and characterized. Limited DNA sequence analysis of random C. parvum clones confirms suggestions that the genome is quite AT-rich. The DNA sequence of random lambda ZAP fusion proteins has identified a potential ATPase, a structural protein and a DNA-binding protein.

  3. Evaluating the transport of bacillus subtilis spores as a potential surrogate for Cryptosporidium parvum Oocysts

    Science.gov (United States)

    The USEPA has recommended the use of aerobic spores as an indicator for Cryptosporidium oocysts when determining groundwater under the direct influence of surface water. Surface properties, interaction energies, transport, retention, and release behavior of B. subtilis spores were measured over a r...

  4. Modellering van de verwijdering van Cryptosporidium parvum,Giardia lamblia en enterovirussen in spaarbekkens

    NARCIS (Netherlands)

    Ratsak CH; MGB

    1996-01-01

    De protozoa Cryptosporidium en Giardia hebben de laatste decennia in de Verenigde Staten en in Groot-Brittannie een aanzienlijk aantal grote en kleine epidemieen van maag-darminfecties via drinkwater veroorzaakt. Wiskundige modellen kunnen worden gebruikt om de verwijdering van pathogene micro-o

  5. Modellering van de verwijdering van Cryptosporidium parvum,Giardia lamblia en enterovirussen in spaarbekkens

    NARCIS (Netherlands)

    Ratsak CH; MGB

    1996-01-01

    De protozoa Cryptosporidium en Giardia hebben de laatste decennia in de Verenigde Staten en in Groot-Brittannie een aanzienlijk aantal grote en kleine epidemieen van maag-darminfecties via drinkwater veroorzaakt. Wiskundige modellen kunnen worden gebruikt om de verwijdering van pathogene

  6. Cryptosporidium infections in children in Durban

    African Journals Online (AJOL)

    1991-03-16

    Mar 16, 1991 ... diarrhoea were found to be passing Cryptosporidium oocysts compared with only 2,4% ... this increase in morbidity and mortality in Cryptosporidium- associated .... taminated water.34 It is possible that contamination of rivers.

  7. Removal of Cryptosporidium parvum oocysts in low quality water using Moringa oleifera seed extract as coagulant

    DEFF Research Database (Denmark)

    Petersen, Heidi Huus; Petersen, T. B.; Enemark, Heidi

    2016-01-01

    The use of different types of low quality water for irrigation in agriculture is common practice in many countries due to limited freshwater resources. Pathogens may contaminate fruit and vegetables when feces contaminated water is used for irrigation or postharvest processing. A laboratory study...... demonstrate that MO seed extract may be used by farmers for treatment of different types of surface water prior to irrigation use. Yet, adding MO seed extract to the low quality water did not successfully remove all oocyst. However, treatment of wastewater with MO seed extract significantly improved the water...... quality with regard to number of oocysts present and turbidity of the water. Further experiments with addition of higher concentrations of MO are needed to establish whether MO seed extract can be used to obtain safe irrigation water free of C. parvum oocysts and other protozoan parasites....

  8. Cryptosporidium cell culture infectivity assay design.

    Science.gov (United States)

    King, B J; Keegan, A R; Robinson, B S; Monis, P T

    2011-05-01

    Members of the genus Cryptosporidium, which cause the gastrointestinal disease cryptosporidiosis, still represent a significant cause of water-borne disease worldwide. While intensive efforts have been invested in the development of techniques for parasite culture, in vitro growth has been hampered by a number of factors including low levels of infectivity as well as delayed life-cycle development and poor synchronicity. In this study we examined factors affecting the timing of contact between excysted sporozoites and target host cells and the subsequent impact of this upon the establishment of infection. We demonstrate that excystation rate impacts upon establishment of infection and that in our standard assay format the majority of sporozoites are not close enough to the cell monolayer when they are released from the oocyst to successfully establish infection. However, this can be easily overcome by centrifugation of oocysts onto the cell monolayer, resulting in approximately 4-fold increases in sporozoite attachment and subsequent infection. We further demonstrate that excystation procedures can be tailored to control excystation rate to match the assay end purpose and that excystation rate can influence data interpretation. Finally, the addition of both a centrifugation and washing step post-sporozoite attachment may be appropriate when considering the design of in vitro culture experiments for developmental analysis and stage-specific gene expression as this appears to increase the synchronicity of early developmental stages.

  9. Role of collector alternating charged patches on transport of Cryptosporidium parvum oocyst in a patchwise charged heterogeneous micromodel

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Yuanyuan; Zhang, Changyong; Hu, Dehong; Kuhlenschmidt, Mark S.; Kuhlenschmidt, Theresa B.; Mylon, Steven E.; Kong, Rong; Bhargava, Rohit; Nguyen, Thanh H.

    2013-02-04

    The role of collector surface charge heterogeneity on transport of Cryptosporidium parvum oocyst and carboxylate microsphere in 2-dimensional micromodels was studied. The cylindrical silica collectors within the micromodels were coated with 0, 10, 20, 50 and 100% Fe2O3 patches. The experimental values of average single collector removal efficiencies (η) of the Fe2O3 patches and on the entire collectors were determined. In the presence of significant (>3500 kT) Derjaguin–Landau–Verwey–Overbeek (DLVO) energy barrier between the microspheres and the silica collectors at pH 5.8 and 8.1, the values of η determined for Fe2O3 patches were significantly less (p < 0.05, t-test) than that obtained for collectors coated entirely with Fe2O3. However, η on Fe2O3 patches for microspheres at pH 4.4 and for oocysts at pH 5.8 and 8.1, where the DLVO energy barrier was relatively small (ca. 200-360 kT), were significantly greater (p < 0.05, t-test) than that on the collectors coated entirely with Fe2O3. The dependence of η determined for Fe2O3 patches on the DLVO energy barrier indicated the importance of periodic favorable and unfavorable electrostatic interactions between colloids and collectors with alternating Fe2O3 and silica patches. Differences between experimentally determined η and that predicted by a patchwise geochemical heterogeneous model was observed, but can be explained by the model’s lack of consideration for the spatial distribution of charge heterogeneity on the collector surface and colloid migration on patchwise heterogeneous collectors.

  10. Cryptosporidium varanii infection in leopard geckos (Eublepharis macularius in Argentina

    Directory of Open Access Journals (Sweden)

    A. Dellarupe

    2016-06-01

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

  11. Molecular characterization of Cryptosporidium spp. from HIV infected patients from an urban area of Brazil Caracterização molecular de Cryptosporidium spp. de pacientes de área urbana do Brasil infectados por HIV

    Directory of Open Access Journals (Sweden)

    Patrícia de Lucca

    2009-12-01

    Full Text Available Cryptosporidium spp. are important cause of enteric disease in humans, but may also infect animals. This study describes the relative frequency of several Cryptosporidium species found in human specimens from HIV infected patients in the São Paulo municipality obtained from January to July 2007. Sequence analysis of the products of nested-PCR based on small subunit rRNA and Cryptosporidium oocyst wall protein coding genes revealed 17 (63.0% isolates of C. hominis, four (14.8% C. parvum, five (18.5% C. felis and one (3.7% C. canis. These findings suggest that, in urban environments of Brazil, the cat adapted C. felis may play a potential role in the zoonotic transmission of cryptosporidiosis whereas the anthroponotic transmission of cryptosporidiosis caused by C. hominis seems to predominate.Cryptosporidium spp. são importantes causas de doenças entéricas em humanos, mas podem também ser encontrados em animais. O presente estudo descreve a frequência relativa de diversas espécies de Cryptosporidium em amostras de humanos da cidade de São Paulo, Brasil, obtidas de janeiro a julho de 2007. Análises de sequências de produtos de nested PCR direcionadas ao genes codificadores da menor unidade ribosomal e da proteina de parede de oocistos revelaram 17 (63,0% isolados de C. hominis, quatro (14,8% C. parvum, cinco (18,5% C. felis, e um (3,7% C. canis. Estes resultados sugerem que, em ambientes urbanos no Brasil, o genótipo adaptado ao gato pode desempenhar potencial papel na transmissão zoonótica de criptosporidiose, enquanto a transmissão antroponótica da criptosporidiose causada pelo C. hominis parece predominar.

  12. Comparison of different solar reactors for household disinfection of drinking water in developing countries: evaluation of their efficacy in relation to the waterborne enteropathogen Cryptosporidium parvum.

    Science.gov (United States)

    Gómez-Couso, H; Fontán-Sainz, M; Navntoft, C; Fernández-Ibáñez, P; Ares-Mazás, E

    2012-11-01

    Solar water disinfection (SODIS) is a type of treatment that can significantly improve the microbiological quality of drinking water at household level and therefore prevent waterborne diseases in developing countries. Cryptosporidium parvum is an obligate protozoan parasite responsible for the diarrhoeal disease cryptosporidiosis in humans and animals. Recently, this parasite has been selected by the WHO as a reference pathogen for protozoan parasites in the evaluation of household water treatment options. In this study, the field efficacy of different static solar reactors [1.5 l transparent plastic polyethylene terephthalate (PET) bottles as well as 2.5 l borosilicate glass and 25 l methacrylate reactors fitted with compound parabolic concentrators (CPC)] for solar disinfection of turbid waters experimentally contaminated with C. parvum oocysts was compared. Potential oocyst viability was determined by inclusion/exclusion of the fluorogenic vital dye propidium iodide. The results demonstrate that static solar reactors fitted with CPCs are an excellent alternative to the conventional SODIS method with PET bottles. These reactors improved the efficacy of the SODIS method by enabling larger volumes of water to be treated and, in some cases, the C. parvum oocysts were rendered totally unviable, minimising the negative effects of turbidity.

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

    Science.gov (United States)

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

    1997-01-01

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

  14. Cryptosporidium infection in children with cancer undergoing chemotherapy: how important is the prevention of opportunistic parasitic infections in patients with malignancies?

    Science.gov (United States)

    Berahmat, Reza; Mahami-Oskouei, Mahmoud; Rezamand, Azim; Spotin, Adel; Aminisani, Nayyereh; Ghoyounchi, Roghayeh; Madadi, Solmaz

    2017-07-20

    Cryptosporidiosis is a relatively uncommon disease in healthy individuals but could be potentially worrisome in immunocompromised patients. This study aimed to evaluate Cryptosporidium infection in children with cancer undergoing chemotherapy. A case-control study was conducted in 132 children with cancer undergoing chemotherapy and 132 non-cancer controls. The modified Ziehl-Neelsen (MZN) staining and polymerase chain reaction methods were used for the detection of Cryptosporidium parasite. All positive isolates were sequenced for phylogenetic analysis. Statistical analysis was performed using the SPSS version 16 and Fisher exact test. The rate of cryptosporidiosis in children with cancer undergoing chemotherapy was 3.8%, which was higher than that of the control group. Other intestinal parasites detected in patients with cancer included Giardia lamblia (3%), Entamoeba coli (1.5%), and Chilomastix mesnili (0.8%). In the control group, only two (1.5%) cases were positive for G. lamblia. No significant difference was observed between the gender, age, residency, contact with domestic animals, stool appearance, neutropenia, chemotherapy period, and type of malignancy with regard to cryptosporidiosis. Phylogenetic analysis revealed that Cryptosporidium parvum isolates in this study relied on a branch that represents similar sequences from Iran and other countries. Although the rate of Cryptosporidium infection was relatively higher in children with cancer undergoing chemotherapy compared to the control group, any statistically significant difference has not been found between them. These findings should not be contrary to the need for healthcare to prevent opportunistic parasitic infections in malignant and immunocompromised patients.

  15. GIS-based analysis of the fate of waste-related pathogens Cryptosporidium parvum, Giardia lamblia and Escherichia coli in a tropical canal network.

    Science.gov (United States)

    Diallo, Mamadou B C; Anceno, Alfredo J; Tawatsupa, Benjawan; Tripathi, Nitin K; Wangsuphachart, Voranuch; Shipin, Oleg V

    2009-03-01

    Urban canals play a major socio-economic role in many tropical countries and, particularly, Thailand. One of the overlooked functions that they perform is a significant attenuation of waste-related pathogens posing considerable health risk, as well as pollution attenuation in general. The study dealt with a comparison of three canals receiving: (i) municipal, (ii) mainly industrial and (iii) mainly agricultural wastewater, listed in order of progressively decreasing organic loading. The occurrence and fate of waterborne Cryptosporidium parvum, Giardia lamblia and Escherichia coli were monitored in the canals by both real-time PCR and conventionally for 12 months. The pathogens are etiological agents of an estimated 38% and 47% of diarrhea cases worldwide and in Thailand, respectively. The geographic information system (GIS) was used to evaluate and map point and, particularly, non-point pollution sources which allowed differentiating the canal sections in terms of predominant pathogen sources. The flowthrough canals, which can be viewed as waste stabilization ponds, were found to be efficiently removing the pathogens at the following generalized specific rates: 0.3 (C. parvum), 1.2 (G. lamblia), 1.8 (E. coli) log10/km.d in the dry season. The rates decreased in the rainy season for E. coli and G. lamblia, but increased for C. parvum which indicated different removal mechanisms. Data suggest that E. coli and G. lamblia were mainly removed through sedimentation and sunlight (UV) irradiation, while the likely mechanism for C. parvum was predation. Overall, the specific pathogen removal rates positively correlated with the canal organic loading rates in the rainy season. As an important result, an estimate of the municipal pollution mitigation by over 2280 km canals in the Greater Bangkok suggests that concomitant to the pathogens at least 36-95 tons of BOD5 is being removed daily, thereby saving the receiving Chao Phraya River and Bight of Bangkok, by far exceeding

  16. Prevalence and abundance of Cryptosporidium parvum and giardia spp. in wild rural rodents from the Mazury Lake District region of Poland.

    Science.gov (United States)

    Bajer, A; Bednarska, M; Pawełczyk, A; Behnke, J M; Gilbert, F S; Sinski, E

    2002-07-01

    Prevalence and abundance of Cryptosporodium parvum and Giardia spp. were studied in 3 species of rodents from forests and abandoned agricultural fields in N.E. Poland (Clethrionomys glareolus n = 459; Microtus arvalis n = 274; Apodemus flavicollis n = 209). Overall prevalence was consistently higher in the voles compared with A. flavicollis (70.6, 73.0 and 27.8% respectively for C. parvum and 93.9, 96.3 and 48.3% respectively for Giardia spp.). Prevalence and abundance of infection also varied markedly across 3 years with 1998 being a year of higher prevalence and abundance with both species. Fewer older animals (especially C. glareolus and M. arvalis) carried infection with C. parvum and infections in these animals were relatively milder. Although seasonal differences were significant, no consistent pattern of changes was apparent. Host sex did not influence prevalence or abundance of infection with C. parvum, but made a small contribution to a 4-way interaction (in 5-way ANOVA) with other factors in the case of Giardia spp. The 2 species co-occurred significantly and in animals carrying both parasites there was a highly significant positive correlation between abundance of infection with each, even with between-year, seasonal, host age, sex and species differences taken into account. Quantitative associations were confined to the 2 vole species in the study. These results are discussed in relation to the importance of wild rodents as reservoir hosts and sources of infection for local human communities.

  17. A recombinant DNA vaccine encoding C. andersoni oocyst wall protein induces immunity against experimental C. parvum infection.

    Science.gov (United States)

    Zheng, Jun; Ren, Wenzhi; Pan, Qingshan; Wang, Qiuyue; Elhag, I A Elfaki; Li, Jianhua; Li, Mingying; Gong, Pengtao; Liu, Yingli; Zhang, Xichen

    2011-06-30

    Cryptosporidium andersoni parasited in the abomasum has been demonstrated as a cause of reduction of milk production in dairy cow. In this study, a novel chimeric DNA vaccine pVAX1-AB was constructed and the efficacy against Cryptosporidium parvum was determined. BALB/c mice were divided into 3 groups and immunized with DNA vaccine expressing the oocyst wall protein, AB protein of C. andersoni, the recombinant plasmid containing the AB gene, respectively. After inoculation of 1 × 10(6) oocysts of C. parvum, the humoral and cellular immune responses were detected. Experimental results showed that the recombinant plasmid can induce corresponding specific antibody response, simultaneously influenced cellular immune responses, and provided greater protection rate (48.6%) than the other groups. These results indicated that chimeric DNA vaccine has a potential in Cryptosporidium vaccine development.

  18. Morphological structure and staining characteristics of Cryptosporidium parvum oocysts%微小隐孢子虫卵囊的形态结构与染色观察

    Institute of Scientific and Technical Information of China (English)

    陈甫

    2011-01-01

    Objective To observe the morphological structure and staining characteristics of Cryptosporidium parvum oocysts. Methods The morphological structure of C. Parvum oocysts was observed using a high-resolution imaging a-nalysis system and depicted using Adobe Photoshop CS4. Characteristics of C. Parvum oocysts stained by modified acid-fast staining, Giemsa staining, Alcian blue staining, and HE staining were observed. Results C. Parvum oocysts with thin oocyst walls appeared round and elliptical in shape and contain one to four crescent-shaped sporozoites and a round residual body. Individual oocysts were empty. C. Parvum oocysts appeared rose pink with modified acid-fast staining, blue with Giemsa staining, green with Alcian blue staining, and red with HE staining. Conclusion The morphological structure of C. Parvum oocysts varied, and oocysts were most effectively stained with modified acid-fast staining.%目的 研究隐孢子虫卵囊的形态与染色特点.方法 应用图像分析系统,观察隐孢子虫卵囊的形态与内部结构,并应用Adobe Photoshop CS4软件绘制各种卵囊形态的模拟图.卵囊经改良抗酸染色法、姬姆萨染色法、阿利新蓝染色法和苏木精-伊红染色法染色后观察其染色特点.结果 隐孢子虫卵囊呈圆形或椭圆形,外层有一薄的卵囊壁,内部含有0~4个月牙形子孢子和圆点状残留体,个别卵囊呈空泡状.隐孢子虫卵囊经改良抗酸染色、姬姆萨染色、阿利新蓝染色和HE染色后,分别呈现玫瑰红色、蓝色、绿色和红色.结论 隐孢子虫卵囊形态和内部结构多样,改良抗酸染色法染色效果较好.

  19. Molecular identification of Giardia and Cryptosporidium from dogs and cats

    Directory of Open Access Journals (Sweden)

    Sotiriadou Isaia

    2013-01-01

    Full Text Available The aim of the present study was to diagnose the presence of Giardia cysts and Cryptosporidium oocysts in household animals using nested polymerase chain reaction (PCR and sequence analysis. One hundred faecal samples obtained from 81 dogs and 19 cats were investigated. The Cryptosporidium genotypes were determined by sequencing a fragment of the small subunit (SSU rRNA gene, while the Giardia Assemblages were determined through analysis of the glutamate dehydrogenase (GDH locus. Isolates from five dogs and two cats were positive by PCR for the presence of Giardia, and their sequences matched the zoonotic Assemblage A of Giardia. Cryptosporidium spp. isolated from one dog and one cat were both found to be C. parvum. One dog isolate harboured a mixed infection of C. parvum and Giardia Assemblage A. These findings support the growing evidence that household animals are potential reservoirs of the zoonotic pathogens Giardia spp. and Cryptosporidium spp. for infections in humans.

  20. Molecular identification of Giardia and Cryptosporidium from dogs and cats.

    Science.gov (United States)

    Sotiriadou, Isaia; Pantchev, Nikola; Gassmann, Doreen; Karanis, Panagiotis

    2013-01-01

    The aim of the present study was to diagnose the presence of Giardia cysts and Cryptosporidium oocysts in household animals using nested polymerase chain reaction (PCR) and sequence analysis. One hundred faecal samples obtained from 81 dogs and 19 cats were investigated. The Cryptosporidium genotypes were determined by sequencing a fragment of the small subunit (SSU) rRNA gene, while the Giardia Assemblages were determined through analysis of the glutamate dehydrogenase (GDH) locus. Isolates from five dogs and two cats were positive by PCR for the presence of Giardia, and their sequences matched the zoonotic Assemblage A of Giardia. Cryptosporidium spp. isolated from one dog and one cat were both found to be C. parvum. One dog isolate harboured a mixed infection of C. parvum and Giardia Assemblage A. These findings support the growing evidence that household animals are potential reservoirs of the zoonotic pathogens Giardia spp. and Cryptosporidium spp. for infections in humans.

  1. Evaluation of the Solar Water Disinfection Process (SODIS) Against Cryptosporidium parvum Using a 25-L Static Solar Reactor Fitted with a Compound Parabolic Collector (CPC)

    Science.gov (United States)

    Fontán-Sainz, María; Gómez-Couso, Hipólito; Fernández-Ibáñez, Pilar; Ares-Mazás, Elvira

    2012-01-01

    Water samples of 0, 5, and 30 nephelometric turbidity units (NTU) spiked with Cryptosporidium parvum oocysts were exposed to natural sunlight using a 25-L static solar reactor fitted with a compound parabolic collector (CPC). The global oocyst viability was calculated by the evaluation of the inclusion/exclusion of the fluorogenic vital dye propidium iodide and the spontaneous excystation. After an exposure time of 8 hours, the global oocyst viabilities were 21.8 ± 3.1%, 31.3 ± 12.9%, and 45.0 ± 10.0% for turbidity levels of 0, 5, and 30 NTU, respectively, and these values were significantly lower (P 10 times). PMID:22302852

  2. Disinfection and toxicological assessments of pulsed UV and pulsed-plasma gas-discharge treated-water containing the waterborne protozoan enteroparasite Cryptosporidium parvum.

    Science.gov (United States)

    Hayes, Jennifer; Kirf, Dominik; Garvey, Mary; Rowan, Neil

    2013-09-01

    We report for the first time on the comparative use of pulsed-plasma gas-discharge (PPGD) and pulsed UV light (PUV) for the novel destruction of the waterborne enteroparasite Cryptosporidium parvum. It also describes the first cyto-, geno- and ecotoxicological assays undertaken to assess the safety of water decontaminated using PPGD and PUV. During PPGD treatments, the application of high voltage pulses (16 kV, 10 pps) to gas-injected water (N2 or O2, flow rate 2.5L/min) resulted in the formation of a plasma that generated free radicals, ultraviolet light, acoustic shock waves and electric fields that killed ca. 4 log C. parvum oocysts in 32 min exposure. Findings showed that PPGD-treated water produced significant cytotoxic properties (as determined by MTT and neutral red assays), genotoxic properties (as determined by comet and Ames assays), and ecotoxic properties (as determined by Microtox™, Thamnotox™ and Daphnotox™ assays) that are representative of different trophic levels in aquatic environment (pozone (0.8 mg/L) and/or dissociated nitric and nitrous acid that contributed to the observed disinfection and toxicity. Chemical analysis of PPGD-treated water revealed increasing levels of electrode metals that were present at ≤ 30 times the tolerated respective values for EU drinking water. PUV-treated water did not exhibit any toxicity and was shown to be far superior to that of PPGD for killing C. parvum oocysts taking only 90 s of pulsing [UV dose of 6.29 μJ/cm(2)] to produce a 4-log reduction compared to a similar reduction level achieved after 32min PPGD treatment as determined by combined in vitro CaCo-2 cell culture-qPCR.

  3. Comparing the efficacy of chlorine, chlorine dioxide, and ozone in the inactivation of Cryptosporidium parvum in water from Parana State, Southern Brazil.

    Science.gov (United States)

    Pereira, Juliana Tracz; Costa, Adriana Oliveira; de Oliveira Silva, Márcia Benedita; Schuchard, Wagner; Osaki, Silvia Cristina; de Castro, Edilene Alcântara; Paulino, Rosangela Clara; Soccol, Vanete Thomaz

    2008-12-01

    In the present work, assays were performed to compare the efficacy of hypochlorous acid, chlorine dioxide, and ozone in the inactivation of Cryptosporidium oocyst in public water supply from Brazilian South conditions. Experiments were carried out in samples containing 2 x 10(4) oocysts/ml of C. parvum purified from feces of experimentally contaminated calves. An in vitro excystation method was used to evaluate oocysts' viability and to determine the inactivation rates of hypochlorous acid at 2 ppm, chlorine dioxide at 1, 2, and 5 ppm, and ozone at the doses of 0.18, 0.24, 0.36, 0.48, and 1.44 mg/l. By using hypochlorous acid, the maximum inactivation rate obtained was 49.04% after 120 min. Chlorine dioxide at 5 ppm inactivated 90.56% of oocysts after 90 min of contact. Ozone was the most effective product, rendering an inactivation of 100% with the concentration of 24 mg/l. Resistance of Cryptosporidium to the usual disinfectants and the need for more effective water treatments to prevent waterborne diseases in Brazil are discussed in this manuscript.

  4. Solar Radiation Induces Non-Nuclear Perturbations and a False Start to Regulated Exocytosis in Cryptosporidium parvum

    Science.gov (United States)

    King, Brendon J.; Hoefel, Daniel; Ee Wong, Pao; Monis, Paul T.

    2010-01-01

    Stratospheric ozone depletion, climate warming and acidification of aquatic ecosystems have resulted in elevated levels of solar radiation reaching many aquatic environments with an increased deleterious impact on a wide range of living organisms. While detrimental effects on living organisms are thought to occur primarily through DNA damage, solar UV can also damage cellular proteins, lipids and signalling pathways. Cryptosporidium, a member of the eukaryotic phylum Apicomplexa, contain numerous vesicular secretory organelles and their discharge via regulated exocytosis is essential for the successful establishment of infection. Using flow cytometric techniques we demonstrate that solar UV rapidly induces sporozoite exocytosis resulting in a significant reduction in the ability of sporozoites to attach and invade host cells. We found that solar UV induced sporozoite membrane depolarization, resulting in reduced cellular ATP and increased cytosolic calcium. These changes were accompanied by a reduction in the internal granularity of sporozoites, indicative of apical organelle discharge, which was confirmed by analysis of sporozoites with an exocytosis-sensitive dye. The precise timing of apical organelle discharge in the presence of a compatible host cell is critical for sporozoite attachment and invasion. Our results demonstrate for the first time how solar UV radiation can interfere with exocytosis, a fundamental cellular process in all eukaryotic cells. We contend that not only may the forecast increases in solar radiation in both aquatic and terrestrial environments significantly affect members of the Apicomplexa, solar UV-induced membrane depolarizations resulting in cytosolic calcium perturbation may affect a wider range of eukaryotic organisms through antagonistic effects on a myriad of calcium dependant cellular functions. PMID:20668710

  5. Solar radiation induces non-nuclear perturbations and a false start to regulated exocytosis in Cryptosporidium parvum.

    Science.gov (United States)

    King, Brendon J; Hoefel, Daniel; Wong, Pao Ee; Monis, Paul T

    2010-07-23

    Stratospheric ozone depletion, climate warming and acidification of aquatic ecosystems have resulted in elevated levels of solar radiation reaching many aquatic environments with an increased deleterious impact on a wide range of living organisms. While detrimental effects on living organisms are thought to occur primarily through DNA damage, solar UV can also damage cellular proteins, lipids and signalling pathways. Cryptosporidium, a member of the eukaryotic phylum Apicomplexa, contain numerous vesicular secretory organelles and their discharge via regulated exocytosis is essential for the successful establishment of infection. Using flow cytometric techniques we demonstrate that solar UV rapidly induces sporozoite exocytosis resulting in a significant reduction in the ability of sporozoites to attach and invade host cells. We found that solar UV induced sporozoite membrane depolarization, resulting in reduced cellular ATP and increased cytosolic calcium. These changes were accompanied by a reduction in the internal granularity of sporozoites, indicative of apical organelle discharge, which was confirmed by analysis of sporozoites with an exocytosis-sensitive dye. The precise timing of apical organelle discharge in the presence of a compatible host cell is critical for sporozoite attachment and invasion. Our results demonstrate for the first time how solar UV radiation can interfere with exocytosis, a fundamental cellular process in all eukaryotic cells. We contend that not only may the forecast increases in solar radiation in both aquatic and terrestrial environments significantly affect members of the Apicomplexa, solar UV-induced membrane depolarizations resulting in cytosolic calcium perturbation may affect a wider range of eukaryotic organisms through antagonistic effects on a myriad of calcium dependant cellular functions.

  6. Speeding up the solar water disinfection process (SODIS) against Cryptosporidium parvum by using 2.5l static solar reactors fitted with compound parabolic concentrators (CPCs).

    Science.gov (United States)

    Gómez-Couso, H; Fontán-Sainz, M; Fernández-Ibáñez, P; Ares-Mazás, E

    2012-12-01

    Water samples of 0, 5, and 100 nephelometric turbidity units (NTU) spiked with Cryptosporidium parvum oocysts were exposed to natural sunlight in 2.5l static borosilicate solar reactors fitted with two different compound parabolic concentrators (CPCs), CPC1 and CPC1.89, with concentration factors of the solar radiation of 1 and 1.89, respectively. The global oocyst viability was calculated by the evaluation of the inclusion/exclusion of the fluorogenic vital dye propidium iodide and the spontaneous excystation. Thus, the initial global oocyst viability of the C. parvum isolate used was 95.3 ± 1.6%. Using the solar reactors fitted with CPC1, the global viability of oocysts after 12h of exposure was zero in the most turbid water samples (100 NTU) and almost zero in the other water samples (0.3 ± 0.0% for 0 NTU and 0.5 ± 0.2% for 5 NTU). Employing the solar reactors fitted with CPC1.89, after 10h exposure, the global oocyst viability was zero in the non-turbid water samples (0 NTU), and it was almost zero in the 5 NTU water samples after 8h of exposure (0.5 ± 0.5%). In the most turbid water samples (100 NTU), the global viability was 1.9 ± 0.6% after 10 and 12h of exposure. In conclusion, the use of these 2.5l static solar reactors fitted with CPCs significantly improved the efficacy of the SODIS technique as these systems shorten the exposure times to solar radiation, and also minimize the negative effects of turbidity. This technology therefore represents a good alternative method for improving the microbiological quality of household drinking water in developing countries. Copyright © 2012 Elsevier B.V. All rights reserved.

  7. Cryptosporidium enteritis

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/article/000617.htm Cryptosporidium enteritis To use the sharing features on this page, please enable JavaScript. Cryptosporidium enteritis is an infection of the small intestine that ...

  8. Comparison of transport and attachment behaviors of Cryptosporidium parvum oocysts and oocyst-sized microspheres being advected through three minerologically different granular porous media.

    Science.gov (United States)

    Mohanram, Arvind; Ray, Chittaranjan; Harvey, Ronald W; Metge, David W; Ryan, Joseph N; Chorover, Jon; Eberl, D D

    2010-10-01

    In order to gain more information about the fate of Cryptosporidium parvum oocysts in tropical volcanic soils, the transport and attachment behaviors of oocysts and oocyst-sized polystyrene microspheres were studied in the presence of two soils. These soils were chosen because of their differing chemical and physical properties, i.e., an organic-rich (43-46% by mass) volcanic ash-derived soil from the island of Hawaii, and a red, iron (22-29% by mass), aluminum (29-45% by mass), and clay-rich (68-76% by mass) volcanic soil from the island of Oahu. A third agricultural soil, an organic- (13% by mass) and quartz-rich (40% by mass) soil from Illinois, was included for reference. In 10-cm long flow-through columns, oocysts and microspheres advecting through the red volcanic soil were almost completely (98% and 99%) immobilized. The modest breakthrough resulted from preferential flow-path structure inadvertently created by soil-particle aggregation during the re-wetting process. Although a high (99%) removal of oocysts and microsphere within the volcanic ash soil occurred initially, further examination revealed that transport was merely retarded because of highly reversible interactions with grain surfaces. Judging from the slope of the substantive and protracted tail of the breakthrough curve for the 1.8-μm microspheres, almost all (>99%) predictably would be recovered within ∼4000 pore volumes. This suggests that once contaminated, the volcanic ash soil could serve as a reservoir for subsequent contamination of groundwater, at least for pathogens of similar size or smaller. Because of the highly reversible nature of organic colloid immobilization in this soil type, C. parvum could contaminate surface water should overland flow during heavy precipitation events pick up near-surface grains to which they are attached. Surprisingly, oocyst and microsphere attachment to the reference soil from Illinois appeared to be at least as sensitive to changes in pH as was

  9. An unexpected phosphate binding site in Glyceraldehyde 3-Phosphate Dehydrogenase: Crystal structures of apo, holo and ternary complex of Cryptosporidium parvum enzyme

    Directory of Open Access Journals (Sweden)

    Chattopadhyay Debasish

    2009-02-01

    Full Text Available Abstract Background The structure, function and reaction mechanism of glyceraldehyde 3-phosphate dehydrogenase (GAPDH have been extensively studied. Based on these studies, three anion binding sites have been identified, one 'Ps' site (for binding the C-3 phosphate of the substrate and two sites, 'Pi' and 'new Pi', for inorganic phosphate. According to the original flip-flop model, the substrate phosphate group switches from the 'Pi' to the 'Ps' site during the multistep reaction. In light of the discovery of the 'new Pi' site, a modified flip-flop mechanism, in which the C-3 phosphate of the substrate binds to the 'new Pi' site and flips to the 'Ps' site before the hydride transfer, was proposed. An alternative model based on a number of structures of B. stearothermophilus GAPDH ternary complexes (non-covalent and thioacyl intermediate proposes that in the ternary Michaelis complex the C-3 phosphate binds to the 'Ps' site and flips from the 'Ps' to the 'new Pi' site during or after the redox step. Results We determined the crystal structure of Cryptosporidium parvum GAPDH in the apo and holo (enzyme + NAD state and the structure of the ternary enzyme-cofactor-substrate complex using an active site mutant enzyme. The C. parvum GAPDH complex was prepared by pre-incubating the enzyme with substrate and cofactor, thereby allowing free movement of the protein structure and substrate molecules during their initial encounter. Sulfate and phosphate ions were excluded from purification and crystallization steps. The quality of the electron density map at 2Å resolution allowed unambiguous positioning of the substrate. In three subunits of the homotetramer the C-3 phosphate group of the non-covalently bound substrate is in the 'new Pi' site. A concomitant movement of the phosphate binding loop is observed in these three subunits. In the fourth subunit the C-3 phosphate occupies an unexpected site not seen before and the phosphate binding loop remains in

  10. Comparison of transport and attachment behaviors of Cryptosporidium parvum oocysts and oocyst-sized microspheres being advected through three minerologically different granular porous media

    Science.gov (United States)

    Mohanram, A.; Ray, C.; Harvey, R.W.; Metge, D.W.; Ryan, J.N.; Chorover, J.; Eberl, D.D.

    2010-01-01

    In order to gain more information about the fate of Cryptosporidium parvum oocysts in tropical volcanic soils, the transport and attachment behaviors of oocysts and oocyst-sized polystyrene microspheres were studied in the presence of two soils. These soils were chosen because of their differing chemical and physical properties, i.e., an organic-rich (43-46% by mass) volcanic ash-derived soil from the island of Hawaii, and a red, iron (22-29% by mass), aluminum (29-45% by mass), and clay-rich (68-76% by mass) volcanic soil from the island of Oahu. A third agricultural soil, an organic- (13% by mass) and quartz-rich (40% by mass) soil from Illinois, was included for reference. In 10-cm long flow-through columns, oocysts and microspheres advecting through the red volcanic soil were almost completely (98% and 99%) immobilized. The modest breakthrough resulted from preferential flow-path structure inadvertently created by soil-particle aggregation during the re-wetting process. Although a high (99%) removal of oocysts and microsphere within the volcanic ash soil occurred initially, further examination revealed that transport was merely retarded because of highly reversible interactions with grain surfaces. Judging from the slope of the substantive and protracted tail of the breakthrough curve for the 1.8-??m microspheres, almost all (>99%) predictably would be recovered within ~4000 pore volumes. This suggests that once contaminated, the volcanic ash soil could serve as a reservoir for subsequent contamination of groundwater, at least for pathogens of similar size or smaller. Because of the highly reversible nature of organic colloid immobilization in this soil type, C. parvum could contaminate surface water should overland flow during heavy precipitation events pick up near-surface grains to which they are attached. Surprisingly, oocyst and microsphere attachment to the reference soil from Illinois appeared to be at least as sensitive to changes in pH as was observed

  11. [Occurrence of Cryptosporidium spp. infection in antillean manatee (Trichechus manatus)].

    Science.gov (United States)

    Borges, João Carlos Gomes; Alves, Leucio Câmara; Vergara-Parente, Jociery Einhardt; Faustino, Maria Aparecida da Glória; Machado, Erilane de Castro Lima

    2009-01-01

    Cryptosporidiosis is a zoonosis which can affect man and a wide range of domestic and wild animals, mainly immunodeficient individuals. The objective of this paper was reported the occurrence of a Cryptosporidium infection in Antillean manatee. After an unusual behavior of an Antillean manatee kept in captivity at the Centro Mamíferos Aquáticos, ICMBio--FMA, clinical examination and posterior fecal sampling was performed. Fecal samples were examined by the Kinyoun technique, Direct Immunofluorescence Test and also examined by 4',6'-Diamidino-2-Phenylindole (DAPI) staining. At the clinical examination, the animal showed signs of abdominal pain. The results obtained by light and fluorescence microscopy analysis showed the presence of Cryptosporidium spp. oocyst in feces of this manatee.

  12. Genotyping of Cryptosporidium isolates from human clinical cases in Poland.

    Science.gov (United States)

    Bajer, Anna; Bednarska, Małgorzata; Cacciò, Simone M; Wolska-Kuśnierz, Beata; Heropolitanska-Pliszka, Edyta; Bernatowska, Ewa; Wielopolska, Małgorzata; Paziewska, Anna; Welc-faleciak, Renata; Siński, Edward

    2008-06-01

    Cryptosporidium spp. infection is usually self-limited in immunocompetent hosts but can be severe and life threatening in children and in immunocompromised individuals including those with primary or acquired immunodeficiencies. One hundred and three faecal samples were collected from 35 hospitalised patients with different symptoms and tested for the presence of the parasite. Cryptosporidium oocysts were found in four of 35 patients (11.4%) using Ziehl-Neelsen staining of faecal smears and immunofluorescence assay, whereas 12 (34.3%) samples tested positive by nested polymerase chain reaction assay. Cryptosporidium DNA was detected in one bile sample but not in a liver tissue biopsy sample collected from a patient who suffered from sclerosing cholangitis. Sequence analysis of oocyst wall protein and beta-tubulin gene fragments revealed three different parasite species (Cryptosporidium hominis, Cryptosporidium meleagridis and Cryptosporidium parvum) in children with primary immunodeficiencies, whereas only C. parvum was found in immunocompetent individuals and in those with secondary immunodeficiencies. This study has revealed a high prevalence of Cryptosporidium infection in hospitalised patients in Poland and confirmed that molecular techniques enable a more sensitive detection of the parasite.

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

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    Yakhchali M.

    2012-02-01

    Full Text Available 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.

  14. Identification of Cryptosporidium Species in Fish from Lake Geneva (Lac Leman in France.

    Directory of Open Access Journals (Sweden)

    Gabriela Certad

    Full Text Available 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

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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

  17. Occurrence of Giardia and Cryptosporidium in pigs on Prince Edward Island, Canada.

    Science.gov (United States)

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

    2012-02-28

    In a cross-sectional study of 633 pigs from 21 herds on Prince Edward Island, Canada (PEI), the prevalence of infection with Cryptosporidium and Giardia, and the genotypes and species of isolates were determined in order to establish the zoonotic potential of pigs in this region. As determined by direct immunofluorescence microscopy (DFA), 18 herds (86%) and 163 animals (26%; 95% CI: 22-29%) tested positive for Cryptosporidium, while just 3 herds (14%) and 6 animals (1%; 95% CI: 0.4-2%) tested positive for Giardia. Cryptosporidium spp. isolates were detected in 39% (95% CI: 34-44%) of weanlings (1-3 months of age) and 9% (95% CI: 6-13) of sows (>8months of age). Molecular characterization using the 18S rDNA and HSP70 gene fragments revealed the presence of Cryptosporidium sp. pig genotype II, C. suis, C. parvum, and Cryptosporidium sp. mouse genotype. Among the 113 isolates of Cryptosporidium spp. successfully genotyped, pig genotype II (61%) predominated, with C. suis (36%) being the next most prominant isolate. C. parvum (2%; two isolates) and Cryptosporidium sp. mouse genotype (0.9%; one isolate) were only occasionally isolated. The only two Cryptosporidium-positive genotyped isolates from sows included one each of C. suis and Cryptosporidium sp. pig genotype II. All but one of the six Giardia positive isolates were detected in weanling pigs. None of the Giardia-positive isolates was amenable to PCR. This study demonstrates that Cryptosporidium spp. are highly prevalent in pigs on PEI, Canada, are found mostly in weanlings (1-3 months of age). Furthermore, the pigs are primarily infected by the host-specific genotypes and species, Cryptosporidium sp. pig genotype II and C. suis, whereas the zoonotic C. parvum is rare. Giardia duodenalis is only occasionally found in pigs. These findings suggest that domestic pigs on PEI, Canada, likely do not pose a significant health risk to humans from these parasites.

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

    Science.gov (United States)

    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.

  19. Epidemiology and public health significance of Cryptosporidium isolated from cattle, buffaloes, and humans in Egypt.

    Science.gov (United States)

    Ibrahim, M A; Abdel-Ghany, A E; Abdel-Latef, G K; Abdel-Aziz, S A; Aboelhadid, S M

    2016-06-01

    The epidemiology and public health significance of Cryptosporidium species and genotypes were investigated in Beni-Suef Governorate, Egypt. A total of 610 animal fecal samples (480 from cattle and 130 from buffaloes) beside 290 stool samples from humans were collected in the period between January and December 2014. Based on the microscopic examination, the overall estimated prevalence of Cryptosporidium spp. in cattle, buffaloes, and humans was 10.2, 12.3, and 19 %, respectively. The highest detection rates were in calves less than 2 months of age (17.1 %) and diarrheic animals (13.0 %). Likewise in humans, the highest prevalence of Cryptosporidium was in infants (31.3 %) and diarrheic individuals (21.1 %). The gender distribution in humans denoted that Cryptosporidium was reported more frequently in males (21.7 %) than females (14.5 %). Based on the molecular characterization of Cryptosporidium, Cryptosporidium oocyst wall protein (COWP) and gp60 genes were successfully amplified in 36 out of 50 samples subjected to genotyping. Restriction fragment length polymorphism (RFLP) analysis of the COWP fragments revealed that Cryptosporidium parvum was the only species detected in cattle (12 isolates) and buffaloes (4 isolates), while in humans, the detected species were Cryptosporidium hominis (15 isolates) and C. parvum (5 isolates). Sequence analysis of the gp60 gene identified the subtype IIdA20G1 within C. parvum isolated from both animals and humans. The common occurrence of zoonotic subtypes of C. parvum in cattle and buffaloes highlights the potential role of these animals as significant reservoirs of infection to humans. Also, the presence of C. hominis and C. parvum in humans indicates that both anthroponotic and zoonotic pathways are expected.

  20. The cell biology of cryptosporidium infection.

    Science.gov (United States)

    O'Hara, Steven P; Chen, Xian-Ming

    2011-08-01

    Cryptosporidiosis remains a significant cause of enteric disease worldwide. Basic investigations of host: pathogen interactions have revealed the intricate processes mediating infection. The following summarizes the interactions that mediate infection and the host responses that both permit and ultimately clear the infection.

  1. Comparison of the Triage Micro Parasite Panel and Microscopy for the Detection of Entamoeba histolytica/Entamoeba dispar, Giardia lamblia, and Cryptosporidium parvum in Stool Samples Collected in Kenya

    Directory of Open Access Journals (Sweden)

    Brett Swierczewski

    2012-01-01

    Full Text Available Entamoeba histolytica, Giardia lamblia, and Cryptosporidium parvum are three of the most important parasitic causes of acute diarrhea worldwide. Laboratory diagnosis of these parasites is usually done by ova and parasite examination (O&P examination via microscopy. The sensitivity and specificity of O&P examination varies among laboratories and can be labor intensive and time consuming. The Triage Micro Parasite Panel (BioSite, San Diego, California is an enzyme immunoassay kit that can detect E. histolytica/E. dispar, G. lamblia, and C. parvum simultaneously using fresh or frozen stool. The present study evaluated the Triage Micro Parasite Panel in detecting E. histolytica/E. dispar, G. lamblia, and C. parvum compared to O&P examination in 266 stool samples collected at medical facilities in Kenya. The sensitivity and specificity results for the Triage Micro Parasite Panel were: for E. histolytica/E. dispar: 100%, 100%, G. lamblia: 100%, 100% and C. parvum: 73%, 100%. There was no evidence of cross reactivity using the kit with other parasites identified in the stool specimens. These results indicate that the Triage Micro Parasite Panel is a highly sensitive kit that can be used for screening purposes in large scale studies or outbreak investigations or as a possible alternative to O&P examination.

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

    Directory of Open Access Journals (Sweden)

    Sahatchai Tangtrongsup

    2017-05-01

    Full Text Available 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.

  3. Enterocytozoon bieneusi, giardia, and Cryptosporidium infecting white-tailed deer.

    Science.gov (United States)

    Santin, Monica; Fayer, Ronald

    2015-01-01

    Despite a white-tailed deer (WTD) population in the United States of approximately 32 million animals extremely little is known of the prevalence and species of the protists that infect these animals. This study was undertaken to determine the presence of potential human protist pathogens in culled WTD in central Maryland. Feces from fawns to adults were examined by molecular methods. The prevalence of Enterocytozoon bieneusi, Cryptosporidium, and Giardia was determined by PCR. All PCR-positive specimens were sequenced to determine the species and genotype(s). Of specimens from 80 WTD, 26 (32.5%) contained 17 genotypes of E. bieneusi. Four genotypes were previously reported (I, J, WL4, LW1) and 13 novel genotypes were identified and named DeerEb1-DeerEb13. Genotypes I, J, and LW1 are known to infect humans. Ten (12.5%) specimens contained the Cryptosporidium deer genotype, and one (1.25%) contained Giardia duodenalis Assemblage A. The identification zoonotic G. duodenalis Assemblage A as well as four E. bieneusi genotypes previously identified in humans suggest that WTD could play a role in the transmission of those parasites to humans.

  4. Candidatus Rickettsia andeanae, a spotted fever group agent infecting Amblyomma parvum ticks in two Brazilian biomes.

    Science.gov (United States)

    Nieri-Bastos, Fernanda Aparecida; Lopes, Marcos Gomes; Cançado, Paulo Henrique Duarte; Rossa, Giselle Ayres Razera; Faccini, João Luiz Horácio; Gennari, Solange Maria; Labruna, Marcelo Bahia

    2014-04-01

    Adult ticks of the species Amblyomma parvum were collected from the vegetation in the Pantanal biome (state of Mato Grosso do Sul) and from horses in the Cerrado biome (state of Piauí) in Brazil. The ticks were individually tested for rickettsial infection via polymerase chain reaction (PCR) targeting three rickettsial genes, gltA, ompA and ompB. Overall, 63.5% (40/63) and 66.7% (2/3) of A. parvum ticks from Pantanal and Cerrado, respectively, contained rickettsial DNA, which were all confirmed by DNA sequencing to be 100% identical to the corresponding fragments of the gltA, ompA and ompB genes of Candidatus Rickettsia andeanae. This report is the first to describe Ca. R. andeanae in Brazil.

  5. Candidatus Rickettsia andeanae, a spotted fever group agent infecting Amblyomma parvum ticks in two Brazilian biomes

    Directory of Open Access Journals (Sweden)

    Fernanda Aparecida Nieri-Bastos

    2014-04-01

    Full Text Available Adult ticks of the species Amblyomma parvum were collected from the vegetation in the Pantanal biome (state of Mato Grosso do Sul and from horses in the Cerrado biome (state of Piauí in Brazil. The ticks were individually tested for rickettsial infection via polymerase chain reaction (PCR targeting three rickettsial genes, gltA, ompA and ompB. Overall, 63.5% (40/63 and 66.7% (2/3 of A. parvum ticks from Pantanal and Cerrado, respectively, contained rickettsial DNA, which were all confirmed by DNA sequencing to be 100% identical to the corresponding fragments of the gltA, ompA and ompB genes of Candidatus Rickettsia andeanae. This report is the first to describe Ca. R. andeanae in Brazil.

  6. Prevalence of the protozoan parasite Cryptosporidium on three organic pig farms in Denmark

    DEFF Research Database (Denmark)

    Petersen, Heidi H.; Jianmin, Wang; Mejer, Helena;

    2013-01-01

    Pigs are a potential source of contamination with Cryptosporidium spp., which can lead to infection in humans. Two species C. parvum and C. hominis can cause an acute diarrheal illness in humans, which can become severe in e.g. patients with HIV. The oocyst can survive for long periods in the env...

  7. Evidence for a new species of Cryptosporidium infecting tortoises: Cryptosporidium ducismarci

    Directory of Open Access Journals (Sweden)

    Traversa Donato

    2010-03-01

    Full Text Available Abstract Cryptosporidiosis affects the gastrointestinal and respiratory tract of humans as well as of a wide range of companion, farm, laboratory and wild animals. In the past few years, three independent studies have provided strong evidence for the existence of a distinct Cryptosporidium species affecting tortoises and likely circulating in other reptile species as well. A new Cryptosporidium genotype was firstly detected and genetically characterized in a marginated tortoise in Italy in 2007 and named Cryptosporidium sp. ex Testudo marginata CrIT-20. The phylogenetic analysis of this isolate indicated that this Cryptosporidium was unique and belonged to the intestinal clade. These findings were later on confirmed by the detection of genetic homologies of isolates from a python and a chameleon from Spain and by recent research in the United States. The latter study presented both the occurrence of intestinal lesions in a pancake tortoise and a Russian tortoise and the genetic characterization of the isolates, together with the first pictures of the endogenous stages of Cryptosporidium CrIT-20. Phylogenetic inference based on the sequences representing small subunit of the nuclear ribosomal RNA gene (SSU of these isolates confirmed the pathological findings because this Cryptosporidium was related to the intestinal group and supported previous results in T. marginata from Italy. The present scientific data on the Cryptosporidium CrIT-20 support its classification as a new species of Cryptosporidium causing intestinal diseases in tortoises. Although further morphological (i.e. exogenous stages and biological aspects (i.e. complete host range are yet to be elucidated, it is proposed that this Cryptosporidium is designated Cryptosporidium ducismarci.

  8. 微小隐孢子虫的基因检测实验研究%EXPERIMENTAL STUDY ON GENE DETECTION OF CRYPTOSPORIDIUM PARVUM

    Institute of Scientific and Technical Information of China (English)

    丁慧萍; 卢思奇; 李凤舞; 戎煜; 张西臣; 王凤云

    2002-01-01

    目的探讨聚合酶链反应(PCR)检测微小隐孢子虫(Cryptosporidium parvum)感染的特异性和敏感性. 方法用Sheather's蔗糖梯度离心法从微小隐孢子虫感染者粪便标本中分离纯化卵囊.用酚-氯仿法抽提卵囊总DNA.针对本虫18S rRNA基因片段设计1对引物,对模板DNA进行PCR扩增,同时以蓝氏贾第鞭毛虫(Giardia lamblia)、溶组织内阿米巴(Entamoeba histolytica)、阴道毛滴虫(Trichomonas vaginalis)、刚地弓形虫(Toxoplasma gondii)、日本血吸虫(Schistosoma japonicum)和旋毛虫(Trichinella spiralis),以及人体血细胞等DNA样本为对照.用琼脂糖电泳和溴乙腚染色对PCR产物进行检测. 结果仅微小隐孢子虫DNA被扩增出一 500 bp的DNA片段.其它对照DNA样本均未出现扩增反应. 结论 PCR对隐孢子虫检测的特异性可高达100%,敏感性也很强,可检测到20 pg微小隐孢子虫卵囊的DNA.表明本实验建立的检测微小隐孢子虫的PCR方法有效.

  9. Influence of organic carbon loading, sediment associated metal oxide content and sediment grain size distributions upon Cryptosporidium parvum removal during riverbank filtration operations, Sonoma County, CA

    Science.gov (United States)

    Metge, D.W.; Harvey, R.W.; Aiken, G.R.; Anders, R.; Lincoln, G.; Jasperse, J.

    2010-01-01

    This study assessed the efficacy for removing Cryptosporidium parvum oocysts of poorly sorted, Fe- and Al-rich, subsurface sediments collected from 0.9 to 4.9 and 1.7-13.9??m below land surface at an operating riverbank filtration (RBF) site (Russian River, Sonoma County, CA). Both formaldehyde-killed oocysts and oocyst-sized (3????m) microspheres were employed in sediment-packed flow-through and static columns. The degree of surface coverage of metal oxides on sediment grain surfaces correlated strongly with the degrees of oocyst and microsphere removals. In contrast, average grain size (D50) was not a good indicator of either microsphere or oocyst removal, suggesting that the primary mechanism of immobilization within these sediments is sorptive filtration rather than physical straining. A low specific UV absorbance (SUVA) for organic matter isolated from the Russian River, suggested that the modest concentration of the SUVA component (0.8??mg??L-1) of the 2.2??mg??L-1 dissolved organic carbon (DOC) is relatively unreactive. Nevertheless, an amendment of 2.2??mg??L-1 of isolated river DOC to column sediments resulted in up to a 35.7% decrease in sorption of oocysts and (or) oocyst-sized microspheres. Amendments (3.2????M) of the anionic surfactant, sodium dodecyl benzene sulfonate (SDBS) also caused substantive decreases (up to 31.9 times) in colloid filtration. Although the grain-surface metal oxides were found to have a high colloid-removal capacity, our study suggested that any major changes within the watershed that would result in long-term alterations in either the quantity and (or) the character of the river's DOC could alter the effectiveness of pathogen removal during RBF operations.

  10. [Progress on the application of Cryptosporidium infected animal models and in vitro cultivation].

    Science.gov (United States)

    Yin, Jian-Hai; Shen, Yu-Juan; Cao, Jian-Ping

    2010-10-30

    The genus Cryptosporidium is composed of protozoan parasites that infect epithelial cells in the microvillus border of the gastrointestinal tract of all classes of vertebrates, and cause severe diarrheal disease in a variety of neonatal animals, children and immunocompromised persons. Establishment of Cryptosporidium infected animal models and its in vitro cultivation system have established a good foundation for characterizing life cycle stage, exploring immunological mechanism, developing vaccines, screening and evaluating potential drugs, as well as assessing oocyst inactivation techniques. This paper reviews recent development and application of the Cryptosporidium infected animal models and its in vitro cultivation.

  11. Molecular characterization of Cryptosporidium isolates from pre-weaned calves in western France in relation to age.

    Science.gov (United States)

    Rieux, Anaïs; Paraud, Carine; Pors, Isabelle; Chartier, Christophe

    2013-10-18

    Eighteen pre-weaned female calves from a single beef cattle herd in western France were sampled weekly from birth to 21/2 months of age in order to characterize Cryptosporidium oocyst output. 182 fecal samples were screened for the presence of oocysts after concentration using immunofluorescence analysis. DNA was extracted from positive samples and a PCR-RFLP protocol, with the restriction enzyme SspI and MboII, to amplify the partial SSU rRNA gene was performed. For the subtyping of Cryptosporidium parvum, a gp60 PCR was carried out. All animals excreted oocysts at at least one sampling date and 80% of the calves presented a mild diarrhea at at least one occasion, with no mortality. The prevalence of excretion reached 94% when calves were 17-23 days of age. The mean number of oocysts at the peak of excretion (10-16 days) was 5 × 10(5) oocysts per gram of feces. PCR-RFLP analysis was successful for 61 of 84 positive samples: 14 were identified as C. parvum, 15 as Cryptosporidium bovis, and 22 as Cryptosporidium ryanae. Ten mixed infections with all combinations of these species were also identified. Calves excreted the following Cryptosporidium species: C. parvum between 7 and 27 days of age, C. bovis between 11 and 38 days and C. ryanae from 19 to 72 days. The IIaA15G2R1 zoonotic subtype of C. parvum subtype was the only subtype identified. We observed the presence of different Cryptosporidium species depending on the age of the animals. This study showed that C. parvum and C. bovis can infect beef calf neonates at similar levels of oocyst excretion with or without clinical signs and that C. parvum isolates had zoonotic potential. Copyright © 2013 Elsevier B.V. All rights reserved.

  12. Prevalence and molecular characterization of Cryptosporidium spp. and Giardia duodenalis in dairy cattle in Beijing, China.

    Science.gov (United States)

    Li, Fuhuang; Wang, Haiyan; Zhang, Zhenjie; Li, Junqiang; Wang, Chenrong; Zhao, Jinfeng; Hu, Suhui; Wang, Rongjun; Zhang, Longxian; Wang, Ming

    2016-03-30

    822 fecal samples from cattle in six areas of Beijing were examined with microscopy for Cryptosporidium oocysts and Giardia cysts. The overall infection rates for Cryptosporidium spp. and Giardia duodenalis were 2.55% and 1.09%, respectively. Cryptosporidium was only detected in calves and heifers, whereas G. duodenalis was found in all age groups. Cryptosporidium spp. were characterized with a PCR-restriction fragment length polymorphism analysis and DNA sequence analysis of the small subunit (SSU) rRNA gene. Two Cryptosporidium species were identified: Cryptosporidium parvum (n=12) and Cryptosporidium andersoni (n=9). Six C. parvum isolates were successfully subtyped with the gp60 gene and three subtypes were detected: IIdA19G1 (n=1), IIdA17G1 (n=1), and IIdA15G1 (n=4). Subtype IIdA17G1 is reported for the first time in cattle worldwide. Nine G. duodenalis isolates were analyzed by sequencing the triosephosphate isomerase (tpi) gene, and only G. duodenalis assemblage E was identified. Therefore, the predominance of C. parvum detected in calves was identical to that found in the Xinjiang Uyghur and Ningxia Hui Autonomous Regions, but differed considerably from that in Henan, Heilongjiang, and Shannxi Provinces. In contrast, the predominance of G. duodenalis assemblage E was more or less similar to its predominance in other areas of China or countries. Our findings confirm the unique character of the C. parvum IId subtypes in China. More systematic studies are required to better understand the transmission of Cryptosporidium and G. duodenalis in cattle in China.

  13. Detection and differentiation of Cryptosporidium by real-time polymerase chain reaction in stool samples from patients in Rio de Janeiro, Brazil

    Directory of Open Access Journals (Sweden)

    Roberta Flávia Ribeiro Rolando

    2012-06-01

    Full Text Available This study reports the first genetic characterisation of Cryptosporidium isolates in Brazil using real-time polymerase chain reaction (RT-PCR. A total of 1,197 faecal specimens from children and 10 specimens from human immunodeficiency virus-infected patients were collected between 1999-2010 and screened using microscopy. Forty-eight Cryptosporidium oocyst-positive isolates were identified and analysed using a generic TaqMan assay targeting the 18S rRNA to detect Cryptosporidium species and two other TaqMan assays to identify Cryptosporidium hominis and Cryptosporidium parvum. The 18S rRNA assay detected Cryptosporidium species in all 48 of the stool specimens. The C. parvum TaqMan assay correctly identified five/48 stool samples, while 37/48 stool specimens were correctly amplified in the C. hominis TaqMan assay. The results obtained in this study support previous findings showing that C. hominis infections are more prevalent than C. parvum infections in Brazil and they demonstrate that the TaqMan RT-PCR procedure is a simple, fast and valuable tool for the detection and differentiation of Cryptosporidium species.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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

  16. Molecular characterization of Cryptosporidium spp. in dairy calves from the state of São Paulo, Brazil.

    Science.gov (United States)

    Meireles, Marcelo V; de Oliveira, Fernando P; Teixeira, Weslen Fabrício P; Coelho, William M D; Mendes, Luiz Cláudio N

    2011-09-01

    Cryptosporidiosis is a common protozoan disease observed in a wide range of vertebrate hosts, including ruminants. Cattle can be a potential reservoir of Cryptosporidium spp., leading to environmental contamination with oocysts of zoonotic species. The molecular characterization of Cryptosporidium spp. isolated from cattle from the state of São Paulo, Brazil, was accomplished using nested polymerase chain reaction for amplification of fragments of the 18S rRNA gene and the glycoprotein GP60 gene, following sequencing of amplified fragments. Positivity for Cryptosporidium was found in 10.7% (21/196) of the samples. Four species of Cryptosporidium were identified: C. andersoni, C. bovis, C. parvum subtype IIaA15G2R1, and C. ryanae. To the best of our knowledge, this is the first report of infection by C. ryanae and C. parvum IIaA15G2R1 in cattle from Brazil.

  17. Age-related Infection with Cryptosporidium Species and Genotype in Pigs in China

    Institute of Scientific and Technical Information of China (English)

    YIN Jian Hai; YUAN Zhong Ying; CAI Hui Xia; SHEN Yu Juan; JIANG Yan Yan; ZHANG Jing; WANG Yan Juan; CAO Jian Ping

    2013-01-01

    Objective Pigs, as hosts of zoonotic Cryptosporidium species/genotypes, are domestic animals with public health significance. The present study was to characterize the infection rate and species/genotype of Cryptosporidium in pre-weaned and post-weaned pigs from Shanghai and Shaoxing, China. Methods A total of 208 fecal samples (42 from pre-weaned piglets, and 166 from post-weaned pigs) were examined by nested PCR of the 18S rRNA gene and analyzed by phylogenetic DNA fragment sequencing of secondary PCR products. Results Infection was detected in 79 samples (19/42 pre-weaned piglets, and 60/166 post-weaned pigs). C. suis (14/79) and Cryptosporidium pig genotype II (65/79) were identified; piglets were more susceptible to the former (13/14) and post-weaned pigs to the latter (59/65). Conclusion Infection of Cryptosporidium spp. in pigs was age-specific;piglets were more susceptible to C. suis while pigs were more susceptible to Cryptosporidium pig genotype II. These findings combined with the isolation of the two Cryptosporidium from water suggest that pigs may be a source of zoonotic Cryptosporidium water pollution. Improvements in pig feeding practices, sewage discharge, feces disposal and field worker protection are therefore important to prevent potential public health problems.

  18. Is real-time PCR-based diagnosis similar in performance to routine parasitological examination for the identification of Giardia intestinalis, Cryptosporidium parvum/Cryptosporidium hominis and Entamoeba histolytica from stool samples? Evaluation of a new commercial multiplex PCR assay and literature review.

    Science.gov (United States)

    Laude, A; Valot, S; Desoubeaux, G; Argy, N; Nourrisson, C; Pomares, C; Machouart, M; Le Govic, Y; Dalle, F; Botterel, F; Bourgeois, N; Cateau, E; Leterrier, M; Le Pape, P; Morio, F

    2016-02-01

    Microscopy is the reference standard for routine laboratory diagnosis in faecal parasitology but there is growing interest in alternative methods to overcome the limitations of microscopic examination, which is time-consuming and highly dependent on an operator's skills and expertise. Compared with microscopy, DNA detection by PCR is simple and can offer a better turnaround time. However, PCR performances remain difficult to assess as most studies have been conducted on a limited number of positive clinical samples and used in-house PCR methods. Our aim was to evaluate a new multiplex PCR assay (G-DiaParaTrio; Diagenode Diagnostics), targeting Giardia intestinalis, Cryptosporidium parvum/Cryptosporidium hominis and Entamoeba histolytica. To minimize the turnaround time, PCR was coupled with automated DNA extraction (QiaSymphony; Qiagen). The PCR assay was evaluated using a reference panel of 185 samples established by routine microscopic examination using a standardized protocol including Ziehl-Neelsen staining and adhesin detection by ELISA (E. histolytica II; TechLab). This panel, collected from 12 French parasitology laboratories, included 135 positive samples for G. intestinalis (n = 38), C. parvum/C. hominis (n = 26), E. histolytica (n = 5), 21 other gastrointestinal parasites, together with 50 negative samples. In all, the G-DiaParaTrio multiplex PCR assay identified 38 G. intestinalis, 25 C. parvum/C. hominis and five E. histolytica leading to sensitivity/specificity of 92%/100%, 96%/100% and 100%/100% for G. intestinalis, C. parvum/C. hominis and E. histolytica, respectively. This new multiplex PCR assay offers fast and reliable results, similar to microscopy-driven diagnosis for the detection of these gastrointestinal protozoa, allowing its implementation in routine clinical practice.

  19. Prevalence and molecular characterization of bovine Cryptosporidium in Qazvin province, Iran.

    Science.gov (United States)

    Keshavarz, Akbar; Haghighi, Ali; Athari, Amid; Kazemi, Bahram; Abadi, Alireza; Nazemalhosseini Mojarad, Ehsan

    2009-03-23

    The aim of this study was to determine the prevalence, variability with host age, and the genotypes of species of Cryptosporidium in cattle from 15 dairy farms in Qazvin province, Iran. Fecal samples, collected from 272 cattle during May 2006 to December 2007, were characterized microscopically. Oocysts from 51 positive samples were analyzed using PCR assay of 18S SSU rRNA, restriction fragment length polymorphism (RFLP) and sequencing. We identified 72.6% of the positive samples as Cryptosporidium parvum, 17.7% as Cryptosporidium andersoni, 7.8% as Cryptosporidium bovis and 1.9% as a novel genotype of C. parvum possessing a single mutation on MboII restriction. An infection rate of 19.5% of C. parvum among 174 pre-weaned calves was significantly higher than the 3.1% among 98 post-weaned calves (P<0.0006). This is the first report of C. bovis and the new subgenotype of C. parvum in Iranian cattle.

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

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    Xiao-Xuan Zhang

    2015-01-01

    Full Text Available 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.

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

    Science.gov (United States)

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

    2012-02-28

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

  3. Genotypic characterization and phylogenetic analysis of Cryptosporidium sp. from domestic animals in Brazil.

    Science.gov (United States)

    Huber, F; da Silva, S; Bomfim, T C B; Teixeira, K R S; Bello, A R

    2007-11-30

    The purpose of the present study was the genetic characterization, sequencing and phylogenetic analysis of 18S rDNA sequences of Cryptosporidium isolates obtained from different animal hosts in Brazil. Fecal samples containing Cryptosporidium oocysts were obtained from chickens, ducks, quails, guinea pigs, dairy calves, dogs and cats. For amplification of 18S rDNA sequences the Secondary-PCR product of the extracted DNA from fecal suspension of each studied animal was utilized. The primary genetic characterization of Cryptosporidium sp. was performed using RFLP with the enzymes SspI and VspI. DNA samples were sequenced and subjected to phylogenetic analysis. The results showed C. baileyi infecting two ducks and one quail and C. melagridis infecting one chicken. The sequences obtained from Cryptosporidium sp. infecting guinea pigs were not identified within groups of known Cryptosporidium species. The isolates found parasitizing cats and one dog were diagnosed as C. felis and C. canis, respectively. One isolate of calf origin was identified as C. parvum. The phylogenetic analysis showed clear distribution of isolates between two Cryptosporidium sp. groups according to their gastric or intestinal parasitism. A great genetic distance was observed between C. felis and C. canis from Brazil when compared to the reference sequences obtained from GenBank. The results obtained during this study constitute the first report of rDNA sequences from C. baileyi, C. meleagridis, C. felis, C. canis and C. parvum isolated in Brazil.

  4. Giardia and Cryptosporidium in cetaceans on the European Atlantic coast.

    Science.gov (United States)

    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.

  5. Synthesis and modulation properties of imidazo[4,5-b]pyridin-7-one and indazole-4,7-dione derivatives towards the Cryptosporidium parvum CpABC3 transporter.

    Science.gov (United States)

    Zeinyeh, Waël; Xia, Hexue; Lawton, Philippe; Radix, Sylvie; Marminon, Christelle; Nebois, Pascal; Walchshofer, Nadia

    2010-06-01

    The syntheses of new N-polysubstituted imidazo[4,5-b]pyridine-7-one (IP, 5 and 8a-8f) and indazole-4,7-dione (ID, 9 and 10) derivatives are described. The binding affinity of IP and ID towards the recombinant Nucleotide Binding Domain NBD1 of Cryptosporidium parvum CpABC3 was evaluated by intrinsic fluorescence quenching. IP induced a moderate quenching of the intrinsic fluorescence of H6-NBD1 whereas IDs 9 and 10 showed a binding affinity comparable to the ATP analogue TNP-ATP. In addition, 8d, 8e and 10 were shown to be competitive inhibitors of the ATPase activity, but with low affinity. These compounds could thus act like some flavonoid derivatives, which can partly overlap both the nucleotide-binding site and the adjacent hydrophobic steroid-binding region of mammalian P-glycoproteins.

  6. Cryptosporidium infection in cattle in Ogun state, Nigeria

    African Journals Online (AJOL)

    ADEYEYE

    2014-06-30

    Jun 30, 2014 ... different studies to identify Cryptosporidium spp. (Kaur et al., 2002; Mahdi .... Pakistan Journal of Biological Sciences, .... Empty rectum and caudal abdominal pain with crepitating .... 1.pdf, retrieved 25-05-2012. Anonymous ...

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

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    Catrin E Moore

    2016-07-01

    Full Text Available 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

  8. AIDS-associated diarrhea and wasting in northeast Brazil is associated with subtherapeutic plasma levels of antiretroviral medications and with both bovine and human subtypes of Cryptosporidium parvum

    Directory of Open Access Journals (Sweden)

    Richard K. Brantley

    2003-02-01

    Full Text Available Advanced HIV infection is frequently complicated by diarrhea, disruption of bowel structure and function, and malnutrition. Resulting malabsorption of or pharmacokinetic changes in antiretroviral agents might lead to subtherapeutic drug dosing and treatment failure in individual patients, and could require dose adjustment and/or dietary supplements during periods of diarrheal illness. We determined the plasma levels of antiretroviral medications in patients that had already been started on medication by their physicians in an urban infectious diseases hospital in northeast Brazil. We also obtained blood samples from patients hospitalized for diarrhea or AIDS-associated wasting, and we found reduced stavudine and didanosine levels in comparison with outpatients without diarrhea or wasting who had been treated at the same hospital clinic. There was a predominance of the protozoal pathogens Cryptosporidium and Isospora belli, typical opportunistic pathogens of AIDS-infected humans, in the stool samples of inpatients with diarrhea. We conclude that severe diarrhea and wasting in this population is associated with both protozoal pathogens and subtherapeutic levels of antiretroviral medications.

  9. Natural infections with Cryptosporidium in the endangered spotted souslik (Spermophilus suslicus).

    Science.gov (United States)

    Kloch, Agnieszka; Bajer, Anna

    2012-03-01

    Cryptosporidium is an intestinal protozoan parasite prevalent in a wide range of mammals. Although it has been recorded in many hosts, its impact on endangered species is poorly understood. Here we present a preliminary study of four populations of the highly threatened spotted souslik (Spermophilus suslicus), living in the westernmost part of the species range. The populations inhabit fragmented habitats and suffer from loss of genetic variation. An IFA test revealed that 35.9% of sampled animals (41/114) was infected with Cryptosporidium and none with Giardia. The prevalence and infection intensity differed among the populations. In areas grazed by cattle it was about 3 folds higher, which suggests a possible transmission route. To the authors best knowledge the present study is the first report of Cryptosporidium infections in S. suslicus.

  10. Ureaplasma urealyticum, Ureaplasma parvum, Mycoplasma hominis and Mycoplasma genitalium infections and semen quality of infertile men

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    Rebai Tarek

    2007-11-01

    Full Text Available Abstract Background Genital ureaplasmas (Ureaplasma urealyticum and Ureaplasma parvum and mycoplasmas (Mycoplasma genitalium and Mycoplasma hominis are potentially pathogenic species playing an etiologic role in both genital infections and male infertility. Reports are, however, controversial regarding the effects of these microorganisms infections in the sperm seminological variables. This study aimed at determining the frequency of genital ureplasmas and mycoplasmas in semen specimens collected from infertile men, and at comparing the seminological variables of semen from infected and non-infected men with these microorganisms. Methods A total of 120 semen samples collected from infertile men were investigated. Semen specimens were examined by in-house PCR-microtiter plate hybridization assay for the presence of genital ureaplasmas and mycoplasmas DNA. Semen analysis was assessed according to the guidelines of the World Health Organization. Standard parametric techniques (t-tests and nonparametric techniques (Wilcoxon tests were used for statistical analysis. Results The frequency of genital ureaplasmas and mycoplasmas detected in semen samples of infertile men was respectively 19.2% (23/120 and 15.8% (19/120. The frequency of Ureaplasma urealyticum (15% was higher than that of Mycoplasma hominis (10.8%, Ureaplasma parvum (4.2% and Mycoplasma genitalium (5%. Mixed species of mycoplasmas and ureaplasmas were detected in 6.7% of semen samples. Comparison of the parameters of the standard semen analysis between the male partners of the infertile couples with and without genital ureaplasmas and mycoplasmas infection showed that the presence of Mycoplasma hominis DNA in semen samples is associated with low sperm concentration (p = 0.007 and abnormal sperm morphology (p = 0.03 and a negative correlation between sperm concentration and the detection of Mycoplasma genitalium in semen samples of infertile men (p = 0.05. The mean values of seminal

  11. Maternal Intravenous Treatment with either Azithromycin or Solithromycin Clears Ureaplasma parvum from the Amniotic Fluid in an Ovine Model of Intrauterine Infection

    OpenAIRE

    2014-01-01

    Intrauterine infection with Ureaplasma spp. is strongly associated with preterm birth and adverse neonatal outcomes. We assessed whether combined intraamniotic (IA) and maternal intravenous (IV) treatment with one of two candidate antibiotics, azithromycin (AZ) or solithromycin (SOLI), would eradicate intrauterine Ureaplasma parvum infection in a sheep model of pregnancy. Sheep with singleton pregnancies received an IA injection of U. parvum serovar 3 at 85 days of gestational age (GA). At 12...

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

    Science.gov (United States)

    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.

  13. The survey of Cryptosporidium infection among young children in kindergartens in Anhui province

    Institute of Scientific and Technical Information of China (English)

    Jun Lu; Chaopin Li; Shan Jiang; Song Ye

    2008-01-01

    Objective: To study the Cryptospondium infection among young children in Anhui province, and to explore the best way to diagnose the disease. Methods: Stool specimens of 1204 children were collected; oocysts of Cryptosporidium were identified with auramine O-modified staining, acid-fast staining, safranine T and methylene blue staining, and auramine O-modified acid-fast staining. Results:The detectable rate of Cryptosporidium in four stainings were respectively 2.46%, 1.50%, 1.98% and 3.46%, and there was a higher significant difference in the rate between auramine O-modified acid-fast staining and the others(P < 0.005). The detectable rate was significantly lower in urban children(2.14%, 15/684) than in rural ones(5.19%, 27/520). Boys and girls had similar detectable rate (1.99%, 24/1 204 vs. 1.50%, 18/1 204). Cryptosporidium infection was usually subclinical, and its major clinical features included benign diarrhea, mild abdominal pain and nausea. Conclusion:Cryptospondium infection was relatively common in kindergartens and a higher infection rate was found in rural children. As the majority of the Cryptosporidium infections were subclinical, diagnosis is important although difficult.

  14. EVALUATION OF EPIDEMIOLOGICAL AND HEALTH FACTORS ASSOCIATED WITH INFECTION BY Cryptosporidium spp. IN DOMICILED DOGS

    Directory of Open Access Journals (Sweden)

    Giselle Ramos da Silva

    2016-07-01

    Full Text Available The genus Cryptosporidium consists of protozoa with large reproduction and dissemination capacity. Transmission can occur indirectly by ingestion of water contaminated with viable oocysts or directly between animals, between humans, or between animals and human. In various parts of the world, pets have been cited as potential sources of infection by Cryptosporidium spp. The present study aimed to evaluate the occurrence of natural Cryptosporidium spp. infection in domestic dogs in the city of Recife, Pernambuco State, Brazil, as well as the association of the infection with epidemiological and sanitary data. Stool samples from 97 dogs were analyzed using the techniques of centrifugal sedimentation in formalin-ether with subsequent staining by Kinyoun method. Statistical analysis was performed using the chi-square test of Pearson and Fisher Exact test, with significance level of 5.0%. Positivity was detected in 31.9% (31/97 of samples. It was the first record of infection by Cryptosporidium spp. in dogs from northeastern Brazil. We verified significant association between the rate of infection and veterinary medical care, vaccination, and type of environment in which the dogs were kept. Keywords: canine; cryptosporidiosis; epidemiology; zoonosis.

  15. Cryptosporidium and Giardia as foodborne zoonoses.

    Science.gov (United States)

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

    2007-10-21

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

  16. [Clinical feature of cryptosporidium infection in HIV/AIDS patients with chronic diarrhea].

    Science.gov (United States)

    Jiao, Bing-xin; Wang, Hui-zhu; Liu, Ying; Li, Juan; Guo, Jie; Li, Min; Wan, Gang; Hua, Wen-hao

    2011-10-11

    To investigate the clinical feature of cryptosporidium infection in HIV/AIDS patients with chronic diarrhea. 253 Stool samples were collected from HIV/AIDS patients with chronic diarrhea during Nov.2009 to Dec.2010. The samples were concentrated by Formalin-Ethyl Acetate Sedimentation technique and stained by Modified acid-fast stain (AFS) for the identification of oocysts by microscopy. Divided the cases into three groups according to their CD4 cell counts (AIDS patients was 12.6% in 253 cases. CD4(+) T-lymphocyte counts was related to the infection rates of cryptosporidium, the difference was statistically significant (χ(2) = 10.33, P difference was no statistically significant (P > 0.05). HIV/AIDS patients with chronic diarrhea who progressed during asymptomatic period, pre-AIDS period, AIDS period, had the infection rate of 0(0/7), 21.3% (19/89), 8.3% (13/157) respectively, the difference was statistically significant (χ(2) = 9.822, P HIV patients were diagnosed with enteritis, the infection rate in urban area and rural area was 6.5% (7/104) and 16.8% (25/149) separately, the difference was statistically significant (χ(2) = 5.596, P different age groups, Cryptosporidium infection status were separately 7.3% (4/55), 13.4% (22/164), 17.6% (6/34). Each group's comparative difference was no statistically significant (χ(2) = 2.29, P > 0.05). The infection rate of cryptosporidium and clinical severity of cryptosporidium infection are statistically correlated with CD4(+) T-lymphocyte counts, with AIDS stage, with HIV associated proctitis.

  17. Genes expressed in grapevine leaves reveal latent wood infection by the fungal pathogen Neofusicoccum parvum.

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    Stefan Czemmel

    Full Text Available Some pathogenic species of the Botryosphaeriaceae have a latent phase, colonizing woody tissues while perennial hosts show no apparent symptoms until conditions for disease development become favorable. Detection of these pathogens is often limited to the later pathogenic phase. The latent phase is poorly characterized, despite the need for non-destructive detection tools and effective quarantine strategies, which would benefit from identification of host-based markers in leaves. Neofusicoccum parvum infects the wood of grapevines and other horticultural crops, killing the fruit-bearing shoots. We used light microscopy and high-resolution computed tomography (HRCT to examine the spatio-temporal relationship between pathogen colonization and anatomical changes in stem sections. To identify differentially-expressed grape genes, leaves from inoculated and non-inoculated plants were examined using RNA-Seq. The latent phase occurred between 0 and 1.5 months post-inoculation (MPI, during which time the pathogen did not spread significantly beyond the inoculation site nor were there differences in lesion lengths between inoculated and non-inoculated plants. The pathogenic phase occurred between 1.5 and 2 MPI, when recovery beyond the inoculation site increased and lesion lengths of inoculated plants tripled. By 2 MPI, inoculated plants also had decreased starch content in xylem fibers and rays, and increased levels of gel-occluded xylem vessels, the latter of which HRCT revealed at a higher frequency than microscopy. RNA-Seq and screening of 21 grape expression datasets identified 20 candidate genes that were transcriptionally-activated by infection during the latent phase, and confirmed that the four best candidates (galactinol synthase, abscisic acid-induced wheat plasma membrane polypeptide-19 ortholog, embryonic cell protein 63, BURP domain-containing protein were not affected by a range of common foliar and wood pathogens or abiotic stresses

  18. Prevalence and determinants of Cryptosporidium spp. infection in smallholder dairy cattle in Iringa and Tanga Regions of Tanzania

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    E.S. Swai

    2007-09-01

    Full Text Available The prevalence of Cryptosporidium spp. infection in a cross-sectional study of dairy cattle, from two contrasting dairying regions in Tanzania, were determined by staining smears of faecal samples with the modified Ziehl-Neelsen technique. Of the 1 126 faecal samples screened, 19.7% were positive for Cryptosporidium spp. The prevalence was lower in Tanga Region than in Iringa Region. The prevalence of affected farms was 20% in Tanga and 21% in Iringa. In both regions, the probability of detecting Cryptosporidium oocysts in faeces varied with animal class, but these were not consistent in both regions. In Tanga Region, Cryptosporidium oocysts were significantly more likely to be found in the faeces of milking cows. In Iringa Region, the likelihood that cattle had Cryptosporidium-positive faeces declined with age, and milking cattle were significantly less likely to have Cryptosporidium positive faeces. In this region, 7% of cattle were housed within the family house at night, and this was marginally associated with a higher likelihood that animals had Cryptosporidium-positive faeces. Our study suggests that even though herd sizes are small, Cryptosporidium spp. are endemic on many Tanzanian smallholder dairy farms. These protozoa may impact on animal health and production, but also on human health, given the close associations between the cattle and their keepers. Further studies are required to assess these risks in more detail, and understand the epidemiology of Cryptosporidium spp. in this management system.

  19. Effects of sediment-associated extractable metals, degree of sediment grain sorting, and dissolved organic carbon upon cryptosporidium parvum removal and transport within riverbank filtration sediments, Sonoma County, California

    Science.gov (United States)

    Metge, D.W.; Harvey, R.W.; Aiken, G.R.; Anders, R.; Lincoln, G.; Jasperse, J.; Hill, M.C.

    2011-01-01

    Oocysts of the protozoan pathogen Cryptosporidium parvum are of particular concern for riverbank filtration (RBF) operations because of their persistence, ubiquity, and resistance to chlorine disinfection. At the Russian River RBF site (Sonoma County, CA), transport of C. parvum oocysts and oocyst-sized (3 ??m) carboxylate-modified microspheres through poorly sorted (sorting indices, ??1, up to 3.0) and geochemically heterogeneous sediments collected between 2 and 25 m below land surface (bls) were assessed. Removal was highly sensitive to variations in both the quantity of extractable metals (mainly Fe and Al) and degree of grain sorting. In flow-through columns, there was a log-linear relationship (r2 = 0.82 at p < 0.002) between collision efficiency (??, the probability that colloidal collisions with grain surfaces would result in attachment) and extractable metals, and a linear relationship (r2 = 0.99 at p < 0.002) between ?? and ??1. Collectively, variability in extractable metals and grain sorting accounted for ???83% of the variability in ?? (at p < 0.0002) along the depth profiles. Amendments of 2.2 mg L-1 of Russian River dissolved organic carbon (DOC) reduced ?? for oocysts by 4-5 fold. The highly reactive hydrophobic organic acid (HPOA) fraction was particularly effective in re-entraining sediment-attached microspheres. However, the transport-enhancing effects of the riverine DOC did not appear to penetrate very deeply into the underlying sediments, judging from high ?? values (???1.0) observed for oocysts being advected through unamended sediments collected at ???2 m bls. This study suggests that in evaluating the efficacy of RBF operations to remove oocysts, it may be necessary to consider not only the geochemical nature and size distribution of the sediment grains, but also the degrees of sediment sorting and the concentration, reactivity, and penetration of the source water DOC. ?? 2011 American Chemical Society.

  20. The Structural Basis of Cryptosporidium-Specific IMP Dehydrogenase Inhibitor Selectivity

    Energy Technology Data Exchange (ETDEWEB)

    MacPherson, Iain S.; Kirubakaran, Sivapriya; Gorla, Suresh Kumar; Riera, Thomas V.; D’Aquino, J. Alejandro; Zhang, Minjia; Cuny, Gregory D.; Hedstrom, Lizbeth (BWH); (Brandeis)

    2010-03-29

    Cryptosporidium parvum is a potential biowarfare agent, an important AIDS pathogen, and a major cause of diarrhea and malnutrition. No vaccines or effective drug treatment exist to combat Cryptosporidium infection. This parasite relies on inosine 5{prime}-monophosphate dehydrogenase (IMPDH) to obtain guanine nucleotides, and inhibition of this enzyme blocks parasite proliferation. Here, we report the first crystal structures of CpIMPDH. These structures reveal the structural basis of inhibitor selectivity and suggest a strategy for further optimization. Using this information, we have synthesized low-nanomolar inhibitors that display 10{sup 3} selectivity for the parasite enzyme over human IMPDH2.

  1. Determination of the level of parasitic infection (Cryptosporidium and Giardia of the vegetables marketed in Ilam city

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    Moyad Avazpoor

    2015-01-01

    Full Text Available Background: Infected with intestinal parasites is one of the most important health and economical problems, which could have different effects, such as diarrheal diseases or death associated. The purpose of this study was to determine the level of prevalence of Cryptosporidium and Giardia parasites in the vegetable marketed in Ilam city. Methods: This study was performed on 280 samples of fresh vegetables and lettuce in Ilam. The samples were taken at the level of 500 grams from the places where vegetables and lettuce are sold. Micro liters of each sample was placed on the slide using automatic micropipette, and Logel and Zyl-Nelson stainings were performed in order to identify Cryptosporidium and Giardia. Results: From 200 samples, 54 samples were contaminated to Cryptosporidium oocyte and 13 samples to Giardia cysts. From 80 lettuce samples also 32 samples were contaminated to Cryptosporidium oocyte, and 6 samples contaminated to Giardia cysts. The results showed that the overall infection was 37%. Infection with Giardia cysts was 6.8% and infection with Cryptosporidium oocyte was 30.7%, and Cryptosporidium infection rates in vegetables and lettuce were different. This difference was statistically significant (P<0.05. Conclusion: As a result of this research it is determined that the prevalence of Giardia and Cryptosporidium in Ilam vegetables is significantly higher, and the contamination of lettuce is far greater. Therefore, authorities should be more attentive to the field of education and the control of parasitic diseases.

  2. Molecular and parasitological study of cryptosporidium isolates from cattle in ilam, west of iran.

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    Mahmoud Mahami Oskouei

    2014-09-01

    Full Text Available Cryptosporidiosis is one of the most important parasitic infections in human and animals. This study was designed for survey on the prevalence of Cryptosporidium infection in farms of Ilam, west of Iran, using parasitology method and genotyping by Nested PCR-RFLP.Fecal samples of 217 cattle were collected fresh and directly from the rectum of cattle. All of the samples were examined by microscopic observation after staining with modified Ziehl-Neelsen (MZN. Genomic DNA extracted by using EURx DNA kit. A Nested PCR-RFLP protocol amplifying 825 bp fragment of 18s rRNA gene conducted to differentiate species and genotyping of the isolates using SspI and VspI as restriction enzymes.The prevalence of Cryptosporidium infection in cattle using both methods is 3.68%. Most of the positive cattle were calves under six months. Species diagnosis carried out by digesting the secondary PCR product with SspI that C. parvum generated 3 visible bands of 448, 247 and 106 bp and digested by VspI restriction enzyme generated 2 visible bands of 628 and 104bp. In this investigation all of the positive samples were Cryptosporidium parvum.C. parvum (bovine genotype detected in all positive cattle samples in Ilam, west of Iran. The results of the present study can help for public health care systems to prevention and management of cryptosporidiosis in cattle and the assessment of cattle cryptosporidiosis as a reservoir for the human infection.

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

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    Daniel J Becker

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

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

    Science.gov (United States)

    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.

  5. A Review of Some Parasite Diseases of African Fish Gut Lumen Protozoa, Coccidioses, Cryptosporidium Infections, Haemoprotozoa, Haemosporidia

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    A.A. Ekubo

    2012-06-01

    Full Text Available A review of some parasite diseases of African fish: Gut lumen protozoa, coccidioses, cryptosporidium infections, haemoprotozoa, haemosporidia was carried out from some existing literature to provide fish culturists and the public sector information on some challenges faced in culture fisheries. The Description, taxonomy, diagnosis Life cycles, biology, Epizootiology and control, Protozoans of the gut lumen, Coccidioses, Cryptosporidium Infections, Haemoprotozoa (Haemoflagellates, Haemosporidia - Dactylosoma and Hemogregarines are reviewed in this study.

  6. Drug treatment and novel drug target against Cryptosporidium

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    Gargala G.

    2008-09-01

    Full Text Available Cryptosporidiosis emergence triggered the screening of many compounds for potential anti-cryptosporidial activity in which the majority were ineffective. The outbreak of cryptosporidiosis which occurred in Milwaukee in 1993 was not only the first significant emergence of Cryptosporidium spp. as a major human pathogen but also a huge waterborne outbreak thickening thousands of people from a major city in North America. Since then, outbreaks of cryptosporidiosis are regularly occurring throughout the world. New drugs against this parasite became consequently urgently needed. Among the most commonly used treatments against cryptosporidiosis are paromomycin, and azithromycin, which are partially effective. Nitazoxanide (NTZ’s effectiveness was demonstrated in vitro, and in vivo using several animal models and finally in clinical trials. It significantly shortened the duration of diarrhea and decreased mortality in adults and in malnourished children. NTZ is not effective without an appropriate immune response. In AIDS patients, combination therapy restoring immunity along with antimicrobial treatment of Cryptosporidium infection is necessary. Recent investigations focused on the potential of molecular-based immunotherapy against this parasite. Others tested the effects of probiotic bacteria, but were unable to demonstrate eradication of C. parvum. New synthetic isoflavone derivatives demonstrated excellent activity against C. parvum in vitro and in a gerbil model of infection. Newly synthesized nitroor non nitro- thiazolide compounds, derived from NTZ, have been recently shown to be at least as effective as NTZ against C. parvum in vitro development and are promising new therapeutic agents.

  7. Presence and molecular characterisation of Giardia and Cryptosporidium in alpacas (Vicugna pacos) from Peru.

    Science.gov (United States)

    Gómez-Couso, Hipólito; Ortega-Mora, Luis M; Aguado-Martínez, Adriana; Rosadio-Alcántara, Raúl; Maturrano-Hernández, Lenin; Luna-Espinoza, Luis; Zanabria-Huisa, Víctor; Pedraza-Díaz, Susana

    2012-07-06

    The presence of Giardia and Cryptosporidium was investigated in 274 faecal samples of alpacas (Vicugna pacos) from 12 herds from Peru by immunofluorescence microscopy and PCR amplification and sequencing of fragments of the ssu-rRNA and β-giardin genes from Giardia spp., as well as the ssu-rRNA gene from Cryptosporidium spp. A total of 137 samples (50.0%) were positive for Giardia spp., and 12 samples (4.4%) for Cryptosporidium spp. In ten samples (3.6%), co-infection by both pathogens was found. Herd prevalence was found to be 91.7% (11/12 herds) for Giardia and 58.3% (7/12 herds) for Cryptosporidium. Regarding the age of the animals, although Giardia was detected in animals as young as 1 week, the prevalence increased with age, reaching 80% by 8 weeks. Similarly, the highest percentage of Cryptosporidium detection (20%) was also found in the 8 week-old group. By PCR, 92 of the 274 analysed samples were positive for Giardia. Sequencing of the amplicons showed the existence of Giardia duodenalis assemblage A in 67 samples; G. duodenalis assemblage E in 24 samples; and inconsistent results between the two molecular markers used in a further sample. Cryptosporidium was only detected by PCR in 3 of the 274 samples; Cryptosporidium parvum was identified in two samples and Cryptosporidium ubiquitum in one sample. This study is the first performing molecular characterisation of both parasites in Peruvian alpacas, and the first report of C. ubiquitum in this host. The identification of G. duodenalis assemblage A, C. parvum and C. ubiquitum, suggests that zoonotic transmission of these enteropathogens between alpacas and humans is possible.

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

    Science.gov (United States)

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

    2009-12-01

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

  9. Small rodents as reservoirs of Cryptosporidium spp. and Giardia spp. in south-western Poland.

    Science.gov (United States)

    Perec-Matysiak, Agnieszka; Buńkowska-Gawlik, Katarzyna; Zaleśny, Grzegorz; Hildebrand, Joanna

    2015-01-01

    Cryptosporidium spp. and Giardia spp. have been detected in a range of host species, including rodents. The aim of this study was to determine the distribution of these pathogens and recognition of the reservoir role of rodents in the maintenance of these pathogens in south-western Poland. Additionally, preliminary molecular studies were conducted to elucidate the species and genotypes of Cryptosporidium and Giardia identified in this study. Stool samples (n=266) from A. agrarius, A. flavicollis and M. glareolus, were subjected for analyses. Values of prevalence were 61.7, 68.3 and 68.1%, respectively, for Cryptosporidium spp. and 41.7, 24.4 and 38.4%, respectively, for Giardia spp. There was a statistically significant correlation between host species and Giardia infection where A. agrarius was the species of the highest prevalence. Statistically significant differences were not found for comparisons made for study sites and occurrence of Giardia spp. and Cryptosporidium spp. Due to preliminary nested PCR results, specific amplifications of Cryptosporidium COWP and SSU rRNA genes were obtained for several isolates taken from rodent host species. One isolate recovered from A. agrarius (from a semi-aquatic, urban area) was identified as C. parvum and revealed 100% similarity with sequences obtained from humans. To the best of the knowledge of the authors, this is the first record of the C. parvum zoonotic species from the striped field mouse. Also recorded were the first findings of C. ubiquitum from three small rodent species.

  10. Between-year variation and spatial dynamics of Cryptosporidium spp. and Giardia spp. infections in naturally infected rodent populations.

    Science.gov (United States)

    Bajer, A

    2008-12-01

    Prevalence and abundance of Cryptosporidium spp. and Giardia spp. infections were studied over the 8-year period in 3 species of rodents in N.E. Poland (bank vole Myodes glareolus-1523; yellow-necked mouse Apodemus flavicollis- 638; common vole Microtus arvalis- 419). Prevalence was 53.8, 28.1 and 62.3% respectively for Cryptosporidium spp. and 58.3, 24.4 and 74.2% respectively for Giardia spp. Prevalence and abundance of infection varied markedly across 8 years of the study with 1998 and 2002 being years of higher prevalence and abundance, following changes in the densities of host species. The distribution of intestinal protozoa in forest rodents did not vary in the 3 isolated sites during the 4-year study. In the case of Cryptosporidium, fewer older animals carried infection and infections of the oldest bank and common voles were relatively milder. In the case of Giardia in yellow-necked mice, infections were more common in older age classes (2 and 3). The two species showed significant co-occurrence and in animals carrying both species there was a strong significant positive correlation between abundance of infection with each. These data are discussed in relation to the parasite genotypes identified in this region and in respect of the role of various ecological factors in shaping of intestinal protozoa communities.

  11. Molecular characterization of Cryptosporidium spp. in children from Mexico.

    Science.gov (United States)

    Valenzuela, Olivia; González-Díaz, Mariana; Garibay-Escobar, Adriana; Burgara-Estrella, Alexel; Cano, Manuel; Durazo, María; Bernal, Rosa M; Hernandez, Jesús; Xiao, Lihua

    2014-01-01

    Cryptosporidiosis is a parasitic disease caused by Cryptosporidium spp. In immunocompetent individuals, it usually causes an acute and self-limited diarrhea; in infants, infection with Cryptosporidium spp. can cause malnutrition and growth retardation, and declined cognitive ability. In this study, we described for the first time the distribution of C. parvum and C. hominis subtypes in 12 children in Mexico by sequence characterization of the 60-kDa glycoprotein (GP60) gene of Cryptosporidium. Altogether, 7 subtypes belonging to 4 subtype families of C. hominis (Ia, Ib, Id and Ie) and 1 subtype family of C. parvum (IIa) were detected, including IaA14R3, IaA15R3, IbA10G2, IdA17, IeA11G3T3, IIaA15G2R1 and IIaA16G1R1. The frequency of the subtype families and subtypes in the samples analyzed in this study differed from what was observed in other countries.

  12. Molecular characterization of Cryptosporidium spp. in children from Mexico.

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    Olivia Valenzuela

    Full Text Available Cryptosporidiosis is a parasitic disease caused by Cryptosporidium spp. In immunocompetent individuals, it usually causes an acute and self-limited diarrhea; in infants, infection with Cryptosporidium spp. can cause malnutrition and growth retardation, and declined cognitive ability. In this study, we described for the first time the distribution of C. parvum and C. hominis subtypes in 12 children in Mexico by sequence characterization of the 60-kDa glycoprotein (GP60 gene of Cryptosporidium. Altogether, 7 subtypes belonging to 4 subtype families of C. hominis (Ia, Ib, Id and Ie and 1 subtype family of C. parvum (IIa were detected, including IaA14R3, IaA15R3, IbA10G2, IdA17, IeA11G3T3, IIaA15G2R1 and IIaA16G1R1. The frequency of the subtype families and subtypes in the samples analyzed in this study differed from what was observed in other countries.

  13. Efficacy of chlorine dioxide tablets on inactivation of cryptosporidium oocysts.

    Science.gov (United States)

    Murphy, Jennifer L; Haas, Charles N; Arrowood, Michael J; Hlavsa, Michele C; Beach, Michael J; Hill, Vincent R

    2014-05-20

    The ability of chlorine dioxide (ClO2) to achieve 2-log inactivation of Cryptosporidium in drinking water has been documented. No studies have specifically addressed the effects of ClO2 on C. parvum oocyst infectivity in chlorinated recreational water venues (e.g., pools). The aim of this research was to determine the efficacy of ClO2 as an alternative to existing hyperchlorination protocols that are used to achieve a 3-log inactivation of Cryptosporidium in such venues. To obtain a 3-log inactivation of C. parvum Iowa oocysts, contact times of 105 and 128 min for a solution containing 5 mg/L ClO2 with and without the addition of 2.6 mg/L free chlorine, respectively, were required. Contact times of 294 and 857 min for a solution containing 1.4 mg/L ClO2 with and without the addition of 3.6 mg/L free chlorine, respectively, were required. The hyperchlorination control (21 mg/L free chlorine only) required 455 min for a 3-log inactivation. Use of a solution containing 5 mg/L ClO2 and solutions containing 5 or 1.4 mg/L ClO2 with the addition of free chlorine appears to be a promising alternative to hyperchlorination for inactivating Cryptosporidium in chlorinated recreational water venues, but further studies are required to evaluate safety constraints on use.

  14. Molecular Characterization of Cryptosporidium spp. in Children from Mexico

    Science.gov (United States)

    Valenzuela, Olivia; González-Díaz, Mariana; Garibay-Escobar, Adriana; Burgara-Estrella, Alexel; Cano, Manuel; Durazo, María; Bernal, Rosa M.; Hernandez, Jesús; Xiao, Lihua

    2014-01-01

    Cryptosporidiosis is a parasitic disease caused by Cryptosporidium spp. In immunocompetent individuals, it usually causes an acute and self-limited diarrhea; in infants, infection with Cryptosporidium spp. can cause malnutrition and growth retardation, and declined cognitive ability. In this study, we described for the first time the distribution of C. parvum and C. hominis subtypes in 12 children in Mexico by sequence characterization of the 60-kDa glycoprotein (GP60) gene of Cryptosporidium. Altogether, 7 subtypes belonging to 4 subtype families of C. hominis (Ia, Ib, Id and Ie) and 1 subtype family of C. parvum (IIa) were detected, including IaA14R3, IaA15R3, IbA10G2, IdA17, IeA11G3T3, IIaA15G2R1 and IIaA16G1R1. The frequency of the subtype families and subtypes in the samples analyzed in this study differed from what was observed in other countries. PMID:24755606

  15. Evaluation of gastroscopic biopsy for diagnosis of Cryptosporidium sp. infection in snakes.

    Science.gov (United States)

    Cerveny, Shannon N S; Garner, Michael M; D'Agostino, Jennifer J; Sekscienski, Stacey R; Payton, Mark E; Davis, Michelle R

    2012-12-01

    Cryptosporidium sp. is a protozoal parasite that can cause hypertrophic gastritis, regurgitation, and chronic wasting in snakes. Diagnosis can be challenging because of intermittent shedding of the organism in the feces. The purpose of this study was to evaluate gastroscopic biopsy as a diagnostic method for Cryptosporidium sp. screening during an outbreak at the Oklahoma City Zoo. Twenty-two snakes were immobilized by chamber induction with an inhalant anesthetic. Gross lesions were seen in seven snakes and consisted of hyperemia, thickening of the gastric mucosa as observed by lack of rugal folds, and a cobblestoned appearance. Gastric biopsy was performed for histologic evaluation (n = 21). Hypertrophic gastritis with intralesional cryptosporidia was diagnosed in five animals. Eleven animals died or were euthanized and complete necropsy was used to evaluate diagnostic measurements of performance of gastric biopsy. Sensitivity for gastric biopsy was 71.4% and specificity was 100%. Gastroscopy appears to be a useful diagnostic method for Cryptosporidium infection in snakes and can help distinguish true infection from prey-based cryptosporidia seen in fecal samples.

  16. Development of an IMS-qPCR Method for Detection of Cryptosporidium parvum in Water%IMS-qPCR检测水源性微小隐孢子虫方法的建立

    Institute of Scientific and Technical Information of China (English)

    高姗姗; 吴绍强; 罗静; 王承民; 张敏; 赵宝华; 何宏轩

    2013-01-01

    Objective To develop a detection method for Cryptosporidium parvum oocysts from water samples,which combined immunomagnetic separation (IMS) with Taqman real-time PCR (qPCR).Methods Conditions of separation and enrichment of IMS method by using specific streptavidin magnetic beads coated with monoclonal antibodies Cp23 directed against C.parvum oocysts were optimized.Special primers of PCR and Taqman probes were designed referring to the 18S rDNA gene of C.parvum (GenBank Accession No.AB513881.1).The conserved genes were amplified from genomic DNA of C.parvum,and then cloned into Peasy-T1 vector.Tenfold dilutions of positive plasmids(104-108copy/μl) were used to construct a standard curves by Taqman qPCR.The specificity of the assay was determined using genomic DNA of C.baileyi,Toxoplasma gondii,C.canis and Escherichia coli.The sensitivity of this assay was evaluated by analyzing 10-fold serially dilutions of plasmids ranging from 100 to 108 copy/μl.Both IMS-qPCR assay and indirect immunofluorescent-antibody assay (IFA) were applied to detect 50 water samples collected from the dairy cattle farms in Hebei.Results The optimal incubation concentration and time of antibody Cp23 were 20 ng/ml and 30 min,respectively,and the catching time was 30 min,the recovery rate was more than 95%.PCR product was 272 bp,and identified by restriction enzyme digestion and nucleotide sequencing.There was a good linear relationship between the standard plasmids and Ct value (correlation r2=0.996 1) of the Taqman qPCR.No cross-reactivity was observed with C.baileyi,T.gondii,C.canis and E.coli.The sensitivity of C.parvum-specific assay was 10 copy/μl.Compared with IFA as golden standard method,the specificity and sensitivity of IMS-qPCR for 50 water samples was 100%(18/18) and 93.8%(30/32),respectively.Conclution The IMS-qPCR assay can be used to specifically detect C.parvum oocysts in water samples.%目的 将免疫磁珠分离技术(IMS)和荧光探针定量PCR (qPCR)相结

  17. First molecular characterisation of Cryptosporidium and Giardia from Bubalus bubalis (water buffalo) in Victoria, Australia.

    Science.gov (United States)

    Abeywardena, Harshanie; Jex, Aaron R; von Samson-Himmelstjerna, Georg; Haydon, Shane R; Stevens, Melita A; Gasser, Robin B

    2013-12-01

    We conducted a molecular epidemiological survey of Cryptosporidium and Giardia from Bubalus bubalis (water buffalo) on two extensive farms (450 km apart) in Victoria, Australia. Faecal samples (n=476) were collected from different age groups of water buffalo at two time points (six months apart) and tested using a PCR-based mutation scanning-targeted sequencing-phylogenetic approach, employing markers within the small subunit of ribosomal RNA (designated pSSU) and triose phosphate isomerase (ptpi) genes. Based on pSSU data, Cryptosporidium parvum, Cryptosporidium bovis and Cryptosporidium genotypes 1, 2 (each 99% similar genetically to Cryptosporidium ryanae) and 3 (99% similar to Cryptosporidium suis) were detected in two (0.4%), one (0.2%), 38 (8.0%), 16 (3.4%) and one (0.2%) of the 476 samples tested, respectively. Using ptpi, Giardia duodenalis assemblages A and E were detected in totals of 56 (11.8%) and six (1.3%) of these samples, respectively. Cryptosporidium was detected on both farms, whereas Giardia was detected only on farm B, and both genera were detected in 1.5% of all samples tested. The study showed that water buffaloes on these farms excreted C. parvum and/or G. duodenalis assemblage A, which are consistent with those found in humans, inferring that these particular pathogens are of zoonotic significance. Future work should focus on investigating, in a temporal and spatial manner, the prevalence and intensity of such infections in water buffaloes in various geographical regions in Australia and in other countries.

  18. Evaluation of immunofluorescence microscopy and enzyme-linked immunosorbent assay in detection of Cryptosporidium and Giardia infections in asymptomatic dogs

    DEFF Research Database (Denmark)

    Rimhanen-Finne, R.; Enemark, Heidi L.; Kolehmainen, J.;

    2007-01-01

    The performance of immunofluorescence microscopy (IF) and enzyme-linked immunosorbent assay (ELISA) in canine feces was evaluated. IF and Cryptosporidium ELISA detected 10(5) oocysts/g, while the detection limit for Giardia ELISA was 10(4) cysts/g. The Cryptosporidium ELISA showed 94% specificity...... but only 71% sensitivity. The Giardia ELISA correlated well with IF (sensitivity 100%, specificity 96%) and was capable of detecting animal specific Giardia duodenalis genotypes. Visual interpretation appeared appropriate for assessment of ELISA results. The proportion of positive samples and possible...... zoonotic character of Cryptosporidium and Giardia infections in 150 asymptomatic Finnish dogs from the Helsinki area were studied. The overall proportion of dogs positive for Cryptosporidium was 5% (7/150) and that for Giardia 5% (8/150). In dogs...

  19. [Cryptosporidium sp infections and other intestinal parasites in food handlers from Zulia state, Venezuela].

    Science.gov (United States)

    Freites, Azael; Colmenares, Deisy; Pérez, Marly; García, María; Díaz de Suárez, Odelis

    2009-03-01

    Cryptosporidiosis in food handlers from Venezuela is unknown, being this an important public health problem in immunosuppressed patients. To determine the prevalence of Cryptosporidium sp and other intestinal parasites in food handlers from Zulia State, one hundred nineteen fecal samples were evaluated by wet mount, concentrated according to Ritchie and modified Ziehl-Neelsen staining. Fourteen (11.8%) were positive for Cryptosporidium sp and associated with other protozoosis (P < 0.05), being most frequent Endolimax nana (42.9%). The general prevalence of the intestinal parasitism was 48.7%, emphasizing E. nana (41.2%), followed by Blastocystis hominis (38.7%) and Entamoeba coli (17.6%). The most frequent pathogenic protozoa was Giardia lamblia (13.4%), followed by the complex Entamoeba histolytica/E. dispar (9.2%). 4.1% were positive for intestinal helminthes. The infection by Cryptosporidium sp is frequent in food handlers from Zulia State. Given to the results of this investigation and the nonexistence of studies in this population, is necessary to deepen in the impact of this parasitism in food handlers and the consumers of their products.

  20. [Detection of Cryptosporidium infection among HIV/AIDS patients with chronic diarrhea in Beijing, Henan and Xinjiang of China].

    Science.gov (United States)

    Wang, Hui-zhu; Jiao, Bing-xin; Tian, Jing-hua; Li, Min; Guo, Jie; Liu, Ying; Li, Xing-wang; Wang, Yu-guang

    2011-09-01

    To investigate the Cryptosporidium infection and its epidemiological characteristics in HIV/AIDS patients with chronic diarrhea. Stool samples collected from HIV/AIDS confirmed patients with chronic diarrhea who lived in Beijing, Henan and Xinjiang. Samples were concentrated by Formalin-Ethyl Acetate Sedimentation technique and stained by modified acid-fast stain (AFS) for the identification of oocysts by microscopy. CD4(+)T cells count was performed by Flow Cytometry. The overall infection rate of Cryptosporidium in AIDS patients was 12.6% (32/253). The infection rates of oocysts in the area of Beijing, Henan and Xinjiang were 5.97% (4/67), 16.1% (24/149) and 10.8% (4/37) respectively. The infection rate of oocysts in the urban areas was 6.5% (7/104) while in the countryside it was 16.8% (25/149) and the difference was significantly different. However, there were no any differences discovered between the infection rates on patient's gender or on infection occurred in different seasons. The infectious rates of oocyst in patients on different stages of the disease were also significantly different (P AIDS patients infected by Cryptosporidium were not rarely seen in northern China. The rate of infection was not associated with patient's gender but was associated with patient's living environments. Patients living in the countryside, with lower lever of CD4(+)T cells counts and at the middle/late stage of the disease, Cryptosporidium infection appeared to be high.

  1. Environmental risk factors for Cryptosporidium infection in an island from Western Venezuela

    Directory of Open Access Journals (Sweden)

    Leonor Chacín-Bonilla

    2008-02-01

    Full Text Available Few investigations have been conducted on risk factors for Cryptosporidium infection in communities from developing countries. A study was conducted to determine the prevalence and risk factors for cryptosporidiosis in San Carlos island, Venezuela. A sample of 515 subjects (mean age ± SD: 21.4 ± 17.8 years was surveyed. Single fecal specimens were collected and modified Ziehl-Neelsen carbolfuchsin staining of formalin-ether concentrate stools were examined for identification of the parasite. Infections with Cryptosporidium (67 of 515, 13% were common. Prevalence of the parasite varied among sectors of the community; 34 of 67(50.7% cases of cryptosporidiosis clustered in two sectors with extreme poverty. Variables strongly associated with a higher risk for the infection (p < 0.01 were residing in these sectors versus the remainder, living in a hut or small residence versus a brick or larger house, using an area of backyard rather than a toilet or latrine for defecation, and having contact with soil contaminated with human feces. Crowding was also a risk (p < 0.05. Contact with human feces contaminated-soil may be an important mode of transmission and poverty a predisposing factor for the infection.

  2. Frequency of cryptosporidium infection in children under five years ...

    African Journals Online (AJOL)

    STORAGESEVER

    2010-02-22

    Feb 22, 2010 ... person, contact with infected animal or by ingestion of contaminated food ... evident by correlation between lower CD4 T cells count, risk and severity of ... for biliary symptom and for death within a year after infection (Vakil et ...

  3. Cryptosporidium genotypes in children and calves living at the wildlife or livestock interface of the Kruger National Park, South Africa.

    Science.gov (United States)

    Abu Samra, Nada; Jori, Ferran; Cacciò, Simone M; Frean, John; Poonsamy, Bhavani; Thompson, Peter N

    2016-05-20

    Cryptosporidium infection is one of the most common causes of parasitic diarrhoea worldwide in cattle and humans. In developing countries, human cryptosporidiosis is most prevalent during early childhood and links between zoonotic infection and animal related activities have been demonstrated. This study investigated the prevalence and species/genotype distribution of Cryptosporidium among children (Kruger National Park in South Africa, where interactions between humans and wild and domestic animals are known to occur. Cryptosporidium oocysts were detected in 8/143 stool samples of children recruited within the hospital system (5.6%; 95% CI 2.4%, 10.7%) and in 2/352 faecal samples of calves (0.6%; 95% CI 0.1%, 2.0%) using the modified Ziehl-Neelsen (MZN) staining technique. Microscopy positive samples from children were further analysed by PCR targeting the 18S rRNA gene and identified as Cryptosporidium hominis (3/4) and Cryptosporidium meleagridis (1/4). Regardless of the microscopy outcome, randomly selected samples (n = 36) from calves 0-4 months of age were amplified and sequenced at the 18S rRNA gene using nested PCR. Two calves tested positive (5.6%; 95% CI 1.7%, 18.7%), and revealed the presence of Cryptosporidium parvum and Cryptosporidium bovis. The detection of only two zoonotic species (C. parvum in one calf and C. meleagridis in one child) suggests that zoonotic cryptosporidiosis is not currently widespread in our study area; however, the potential exists for amplification of transmission in an immunocompromised population.

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

    Science.gov (United States)

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

    2012-06-01

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

  5. Ureaplasma parvum serovar 3 multiple banded antigen size variation after chronic intra-amniotic infection/colonization.

    Directory of Open Access Journals (Sweden)

    James W Robinson

    Full Text Available Ureaplasma species are the microorganisms most frequently associated with adverse pregnancy outcomes. The multiple banded antigen (MBA, a surface-exposed lipoprotein, is a key virulence factor of ureaplasmas. The MBA demonstrates size variation, which we have shown previously to be correlated with the severity of chorioamnion inflammation. We aimed to investigate U. parvum serovar 3 pathogenesis in vivo, using a sheep model, by investigating: MBA variation after long term (chronic and short term (acute durations of in utero ureaplasma infections, and the severity of chorioamnionitis and inflammation in other fetal tissues. Inocula of 2 × 10(7 colony-forming-units (CFU of U. parvum serovar 3 (Up or media controls (C were injected intra-amniotically into pregnant ewes at one of three time points: day 55 (69d Up, n = 8; C69, n = 4; day 117 (7d Up, n = 8; C7, n = 2; and day 121 (3d Up, n = 8; C3, n = 2 of gestation (term = 145-150d. At day 124, preterm fetuses were delivered surgically. Samples of chorioamnion, fetal lung, and umbilical cord were: (i snap frozen for subsequent ureaplasma culture, and (ii fixed, embedded, sectioned and stained by haematoxylin and eosin stain for histological analysis. Selected fetal lung clinical ureaplasma isolates were cloned and filtered to obtain cultures from a single CFU. Passage 1 and clone 2 ureaplasma cultures were tested by western blot to demonstrate MBA variation. In acute durations of ureaplasma infection no MBA variants (3d Up or very few MBA variants (7d Up were present when compared to the original inoculum. However, numerous MBA size variants were generated in vivo (alike within contiguous tissues, amniotic fluid and fetal lung, but different variants were present within chorioamnion, during chronic, 69d exposure to ureaplasma infection. For the first time we have shown that the degree of ureaplasma MBA variation in vivo increased with the duration of gestation.

  6. Ureaplasma parvum serovar 3 multiple banded antigen size variation after chronic intra-amniotic infection/colonization.

    Science.gov (United States)

    Robinson, James W; Dando, Samantha J; Nitsos, Ilias; Newnham, John; Polglase, Graeme R; Kallapur, Suhas G; Pillow, J Jane; Kramer, Boris W; Jobe, Alan H; Payton, Diane; Knox, Christine L

    2013-01-01

    Ureaplasma species are the microorganisms most frequently associated with adverse pregnancy outcomes. The multiple banded antigen (MBA), a surface-exposed lipoprotein, is a key virulence factor of ureaplasmas. The MBA demonstrates size variation, which we have shown previously to be correlated with the severity of chorioamnion inflammation. We aimed to investigate U. parvum serovar 3 pathogenesis in vivo, using a sheep model, by investigating: MBA variation after long term (chronic) and short term (acute) durations of in utero ureaplasma infections, and the severity of chorioamnionitis and inflammation in other fetal tissues. Inocula of 2 × 10(7) colony-forming-units (CFU) of U. parvum serovar 3 (Up) or media controls (C) were injected intra-amniotically into pregnant ewes at one of three time points: day 55 (69d Up, n = 8; C69, n = 4); day 117 (7d Up, n = 8; C7, n = 2); and day 121 (3d Up, n = 8; C3, n = 2) of gestation (term = 145-150d). At day 124, preterm fetuses were delivered surgically. Samples of chorioamnion, fetal lung, and umbilical cord were: (i) snap frozen for subsequent ureaplasma culture, and (ii) fixed, embedded, sectioned and stained by haematoxylin and eosin stain for histological analysis. Selected fetal lung clinical ureaplasma isolates were cloned and filtered to obtain cultures from a single CFU. Passage 1 and clone 2 ureaplasma cultures were tested by western blot to demonstrate MBA variation. In acute durations of ureaplasma infection no MBA variants (3d Up) or very few MBA variants (7d Up) were present when compared to the original inoculum. However, numerous MBA size variants were generated in vivo (alike within contiguous tissues, amniotic fluid and fetal lung, but different variants were present within chorioamnion), during chronic, 69d exposure to ureaplasma infection. For the first time we have shown that the degree of ureaplasma MBA variation in vivo increased with the duration of gestation.

  7. Ocorrência de infecção Cryptosporidium spp. em peixe-boi marinho (Trichechus manatus Occurrence of Cryptosporidium spp. infection in antillean manatee (Trichechus manatus

    Directory of Open Access Journals (Sweden)

    João Carlos Gomes Borges

    2009-03-01

    Full Text Available A criptosporidiose constitui-se como uma zoonose que pode afetar o homem e uma ampla variedade de animais domésticos e silvestres, principalmente indivíduos imunodeficientes. O objetivo desse trabalho foi registrar a ocorrência de infecção por Cryptosporidium em peixe-boi marinho. Após ser constatada a mudança de comportamento de um peixe-boi marinho mantido nos oceanários do Centro Mamíferos Aquáticos, ICMBio - FMA, animal foi submetido à exame clínico e, posteriormente, à coleta de amostra fecal. As amostras fecais foram analisadas pela técnica de Kinyoun, teste de imunofluorescência direta e pelo corante 4'.6'-Diamidino-2-Phenilindole (DAPI. No exame clínico, o animal apresentou sinais de desconforto abdominal. Os resultados obtidos nas análises de microscopia de luz e fluorescente revelaram a presença de oocistos de Cryptosporidium nas fezes desse peixe-boi.Cryptosporidiosis is a zoonosis which can affect man and a wide range of domestic and wild animals, mainly immunodeficient individuals. The objective of this paper was reported the occurrence of a Cryptosporidium infection in Antillean manatee. After an unusual behavior of an Antillean manatee kept in captivity at the Centro Mamíferos Aquáticos, ICMBio - FMA, clinical examination and posterior fecal sampling was performed. Fecal samples were examined by the Kinyoun technique, Direct Immunofluorescence Test and also examined by 4'.6'-Diamidino-2-Phenylindole (DAPI staining. At the clinical examination, the animal showed signs of abdominal pain. The results obtained by light and fluorescence microscopy analysis showed the presence of Cryptosporidium spp. oocyst in feces of this manatee.

  8. Cryptosporidium genotypes in children and calves living at the wildlife or livestock interface of the Kruger National Park, South Africa

    Directory of Open Access Journals (Sweden)

    Nada Abu Samra

    2016-03-01

    Full Text Available Cryptosporidium infection is one of the most common causes of parasitic diarrhoea worldwide in cattle and humans. In developing countries, human cryptosporidiosis is most prevalent during early childhood and links between zoonotic infection and animal related activities have been demonstrated. This study investigated the prevalence and species/genotype distribution of Cryptosporidium among children (< 5 years and calves (< 6 months living in a rural farming area adjacent to the Kruger National Park in South Africa, where interactions between humans and wild and domestic animals are known to occur. Cryptosporidium oocysts were detected in 8/143 stool samples of children recruited within the hospital system (5.6%; 95% CI 2.4%, 10.7% and in 2/352 faecal samples of calves (0.6%; 95% CI 0.1%, 2.0% using the modified Ziehl–Neelsen (MZN staining technique. Microscopy positive samples from children were further analysed by PCR targeting the 18S rRNA gene and identified as Cryptosporidium hominis (3/4 and Cryptosporidium meleagridis (1/4. Regardless of the microscopy outcome, randomly selected samples (n = 36 from calves 0–4 months of age were amplified and sequenced at the 18S rRNA gene using nested PCR. Two calves tested positive (5.6%; 95% CI 1.7%, 18.7%, and revealed the presence of Cryptosporidium parvum and Cryptosporidium bovis. The detection of only two zoonotic species (C. parvum in one calf and C. meleagridis in one child suggests that zoonotic cryptosporidiosis is not currently widespread in our study area; however, the potential exists for amplification of transmission in an immunocompromised population.Keywords: Cryptosporidium; children; calves; South Africa; genotyping; GP60 subtyping

  9. COMPARAÇÃO DA EFICIÊNCIA DAS COLORAÇÕES DE ZIEHL- NEELSEN MODIFICADO E SAFRANINA MODIFICADA NA DETECÇÃO DE OOCISTOS DE Cryptosporidium spp. (EUCOCCIDIORIDA, CRYPTOSPORIDIIDAE A PARTIR DE AMOSTRAS FECAIS DE BEZERROS DE 0 A 3 MESES

    Directory of Open Access Journals (Sweden)

    Renata Dias Rodrigues

    2016-01-01

    Full Text Available Bovine cryptosporidiosis is caused by four differents s pecies: C ryptosporidium parvum, Cryptosporidium bovis, Cryptosporidium andersoni and Cryptosporidium ryanae. The species Cryptosporidium parvum (Order: Eucoccidiorida, Family: Cryptosporidiidae is considered of high zoonotic potential and it can infect humans through the elimination of oocysts by both cattle and by humans. The objective of this research was to detect oocysts of the genus Cryptosporidium spp. in fecal contents of calves (75 males and 77 females. We collected 152 stool samples from animals aged between 0 day and 3 months. The material was subjected to modified Ziehl-Neelsen and modified Safranin techniques, the slides were observed in its entire length by optical microscopy to verify the presence of oocysts of this parasitic infections. The results showed 17.1% (26/152 positivity in the samples examined, and the statistical analysis showed no difference between sex and the staining techniques used in this study. We concluded the infection by Cryptosporidium spp. is present in the evaluated properties, but more studies are needed, so that the risk of infection is measured properly and prophylactic measures are implemented.

  10. Intestinal immune response to human Cryptosporidium sp. infection

    Science.gov (United States)

    2008-01-01

    intestinal tissues from AIDS patients with cryptosporidiosis, we did not detect CXCL-8 (90). Finally, CXCL-8 attracts mainly granu - locytes. However, in...cryptosporidiosis but not after reconstitution of immunity. Infect. Immun. 75:481–487. 91. Weinstock, J. V., A. Blum, J. Walder, and R. Walder. 1988. Eosinophils

  11. Parasitic infections in HIV infected individuals: Diagnostic & therapeutic challenges

    OpenAIRE

    Nissapatorn, Veeranoot; Sawangjaroen, Nongyao

    2011-01-01

    After 30 years of the human immunodeficiency virus (HIV) epidemic, parasites have been one of the most common opportunistic infections (OIs) and one of the most frequent causes of morbidity and mortality associated with HIV-infected patients. Due to severe immunosuppression, enteric parasitic pathogens in general are emerging and are OIs capable of causing diarrhoeal disease associated with HIV. Of these, Cryptosporidium parvum and Isospora belli are the two most common intestinal protozoan p...

  12. Comparação entre os métodos de Ziehl-Neelsen modificado e Acid-Fast-Trichrome para a pesquisa fecal de Cryptosporidium parvum e Isospora belli

    Directory of Open Access Journals (Sweden)

    Rigo Carla R.

    2002-01-01

    Full Text Available Devido a crescente importância dos coccídios intestinais (Cryptosporidium, Isospora e Cyclospora como parasitos oportunistas, é fundamental para os laboratórios diferenciar morfologicamente estes protozoários; a técnica de Ziehl-Neelsen modificada (ZNm é amplamente utilizada para este fim; recentemente, foi proposto um novo procedimento, a coloração combinada do ácido tricrômico (Acid-Fast-Trichrome - AFT. O objetivo do presente estudo foi comparar os processos AFT e ZNm para a detecção destes coccídios em amostras fecais de pacientes portadores do vírus VIH. Foram selecionados dois grupos de indivíduos, para inclusão no estudo, segundo a presença (n=60 ou ausência de diarréia (n=60. As amostras de fezes foram coletadas em solução de formalina 10% e os esfregaços fecais preparados i diretamente das fezes e ii após concentração prévia a 500xg (10 minutos, foram submetidos aos diferentes processos de coloração. Considerando-se a positividade por técnica (AFT e ZNm, verificou-se a superioridade do procedimento de ZNm (n=19; 100% dos casos positivos sobre o de AFT (n=8; 42,1%. Ambos possibilitaram a identificação dos 101 casos verdadeiramente negativos. Coccidiose intestinal foi mais frequente entre os pacientes que apresentaram diarréia (26,6% em comparação à positividade observada entre os indíviduos assintomáticos (5% sendo que C. cayetanensis não foi detectada em ambos os grupos. Foi de nosso interesse avaliar a aplicabilidade da técnica AFT para a coloração deste protozoário. Devido à sensibilidade e especificidade obtida neste estudo (100%, conclui-se que o método de ZNm continua sendo o mais indicado para o diagnóstico da criptosporidiose e isosporose, principalmente quando associado ao procedimento de centrífugo-concentração (500xg, 10 minutos. Embora a coloração AFT tenha baixo custo, faz-se necessário o seu aperfeiçoamento pois este procedimento permite o diagnóstico simultâneo dos

  13. Cryptosporidium infection in infancy as a cause of malnutrition

    DEFF Research Database (Denmark)

    Mølbak, Kare; Andersen, M; Aaby, Peter

    1997-01-01

    . A total of 5072 weight and 4264 height measurements was made. There were no tendencies of low weight (P = 0.38) or height (P = 0.16) in children who acquired cryptosporidiosis. Cryptosporidiosis in infancy was accompanied by an estimated weight loss of 392 g (95% CI: 247, 538 g) in boys and 294 g (95% CI......: 109, 479 g) in girls, corresponding to 3.7% and 2.9% of mean weight, respectively, at 2 y of age. No significant catch-up growth covered for this loss in weight. A similar effect in linear growth was shown (P = 0.02). Although it has been suggested that the effect of infections on nutrition is usually...

  14. Diversity of Cryptosporidium species occurring in sheep and goat breeds reared in Poland.

    Science.gov (United States)

    Kaupke, Agnieszka; Michalski, Mirosław M; Rzeżutka, Artur

    2017-03-01

    The aim of this study was molecular identification of Cryptosporidium species and assessment of their prevalence in different breeds of sheep and goat reared in Poland. In addition, the relationship between animal age, breed type, and the frequency of Cryptosporidium infections was determined. Fecal samples from 234 lambs and 105 goat kids aged up to 9 weeks, representing 24 breeds and their cross-breeds were collected from 71 small ruminant farms across Poland. The identification of Cryptosporidium species was performed at the 18 SSU ribosomal RNA (rRNA) and COWP loci followed by subtyping of C. parvum and C. hominis strains at GP60 gene locus. The presence of Cryptosporidium DNA at the 18 SSU rRNA locus was detected in 45/234 (19.2%) lamb feces samples and in 39/105 (37.1%) taken from goats. The following Cryptosporidium species: C. xiaoi, C. bovis, C. ubiquitum, C. parvum, and C. hominis were detected in small ruminants. Infections caused by C. xiaoi were predominant without favoring any tested animal species. Subsequent GP60 subtyping revealed the presence of C. parvum IIaA17G1R1 subtype in sheep and IIdA23G1 subtype in goats. IIdA23G1 subtype was detected in a goat host for the first time. There were no significant differences found in frequency of infections between the age groups ( 0.05) or goat kids (P = 0.06, α > 0.05). In addition, there was no correlation observed between the frequency in occurrence of particular parasite species and breed type in relation to native sheep breeds (F = 0.11; P = 0.990 > 0.05). In the case of goats, more breed-related differences in parasite occurrence were found. The results of this study improve our knowledge on the breed-related occurrence of Cryptosporidium infections in the population of small ruminants reared in Poland.

  15. Molecular characterization of isolates of waterborne Cryptosporidium spp. collected during an outbreak of gastroenteritis in South Burgundy, France.

    Science.gov (United States)

    Dalle, Frédéric; Roz, Pascale; Dautin, Guillaume; Di-Palma, Marc; Kohli, Evelyne; Sire-Bidault, C; Fleischmann, Marie George; Gallay, Anne; Carbonel, Sylvia; Bon, Fabienne; Tillier, Claude; Beaudeau, Pascal; Bonnin, Alain

    2003-06-01

    In September 2001, a waterborne outbreak of gastroenteritis occurred in eastern France. Of 31 fecal samples from symptomatic individuals, 19 tested positive for Cryptosporidium with two PCRs targeting the Hsp70 and the 18S rRNA genes of CRYPTOSPORIDIUM: Sequencing of the PCR fragments produced sequences identical to that of Cryptosporidium parvum genotype 1.

  16. Cryptosporidium proliferans n. sp. (Apicomplexa: Cryptosporidiidae: Molecular and Biological Evidence of Cryptic Species within Gastric Cryptosporidium of Mammals.

    Directory of Open Access Journals (Sweden)

    Martin Kváč

    Full Text Available The morphological, biological, and molecular characteristics of Cryptosporidium muris strain TS03 are described, and the species name Cryptosporidium proliferans n. sp. is proposed. Cryptosporidium proliferans obtained from a naturally infected East African mole rat (Tachyoryctes splendens in Kenya was propagated under laboratory conditions in rodents (SCID mice and southern multimammate mice, Mastomys coucha and used in experiments to examine oocyst morphology and transmission. DNA from the propagated C. proliferans isolate, and C. proliferans DNA isolated from the feces of an African buffalo (Syncerus caffer in Central African Republic, a donkey (Equus africanus in Algeria, and a domestic horse (Equus caballus in the Czech Republic were used for phylogenetic analyses. Oocysts of C. proliferans are morphologically distinguishable from C. parvum and C. muris HZ206, measuring 6.8-8.8 (mean = 7.7 μm × 4.8-6.2 μm (mean = 5.3 with a length to width ratio of 1.48 (n = 100. Experimental studies using an isolate originated from T. splendens have shown that the course of C. proliferans infection in rodent hosts differs from that of C. muris and C. andersoni. The prepatent period of 18-21 days post infection (DPI for C. proliferans in southern multimammate mice (Mastomys coucha was similar to that of C. andersoni and longer than the 6-8 DPI prepatent period for C. muris RN66 and HZ206 in the same host. Histopatologicaly, stomach glands of southern multimammate mice infected with C. proliferans were markedly dilated and filled with necrotic material, mucus, and numerous Cryptosporidium developmental stages. Epithelial cells of infected glands were atrophic, exhibited cuboidal or squamous metaplasia, and significantly proliferated into the lumen of the stomach, forming papillary structures. The epithelial height and stomach weight were six-fold greater than in non-infected controls. Phylogenetic analyses based on small subunit rRNA, Cryptosporidium

  17. Multilocus genotyping of Cryptosporidium hominis associated with diarrhea outbreak in a day care unit in São Paulo Genotipagem de multilocus de Cryptosporidium hominis associado a surto diarréico em creche de São Paulo

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    Elenice Messias do Nascimento Gonçalves

    2006-04-01

    Full Text Available A number of species of Cryptosporidium are associated with diarrhea worldwide. Little data exists regarding the genotypes and species of Cryptosporidium associated with cases of infections in Brazil. PURPOSE: In the present study, we ascertained by molecular methods the species and the genotype of Cryptosporidium sp from a diarrhea outbreak diagnosed in a day care at the Hospital Clínicas, São Paulo University Medical School. MATERIALS AND METHODS: Specific identification and typing of the isolates associated with the outbreak was done by DNA sequencing analysis of fragments amplified by polymerase chain reaction (PCR from 3 different Cryptosporidium loci: the SSUrRNA coding region, the Cryptosporidium oocyst wall protein (COWP gene, and the microsatellite locus 1 (ML1, a tandem GAG-trinucleotide repeat containing substitutions that differentiate the genotypes of Cryptosporidium parvum and Cryptosporidium hominis. RESULTS: A total of 29 positive samples from the outbreak were studied by the molecular methods described. Our study revealed the presence of a single genotype of Cryptosporidium hominis in all samples. CONCLUSION: The molecular analysis reinforced the hypothesis that the transmission of Cryptosporidium hominis during the period the samples were collected occurred in an outbreak pattern, possibly by person-to-person contact through the fecal-oral route. As far as we know, this is the first time that molecular tools have been used to identify the species and the genotype of isolates showing the presence of the ML1 genotype in samples from Brazilian patients.Mundialmente, diferentes espécies de Cryptosporidium estão relacionadas com doenças diarréicas. No Brasil há poucos dados sobre os genótipos das espécies de Cryptosporidium associadas a infecções. OBJETIVO: No presente estudo, caracterizamos, por métodos moleculares, a espécie e o genótipo de Cryptosporidium sp diagnosticado em surto diarréico ocorrido na creche do

  18. Eimeria and Cryptosporidium in Estonian dairy farms in regard to age, species, and diarrhoea.

    Science.gov (United States)

    Lassen, Brian; Viltrop, Arvo; Raaperi, Kerli; Järvis, Toivo

    2009-12-23

    Eimeria and Cryptosporidium are among the most common bovine parasites in the world, but little is known about them in Estonia. Basic field research is needed to gain insight into pathogen dynamics, providing knowledge for veterinarians and research. A survey of 45 Estonian dairy farms in 15 counties was carried out between 2006 and 2007. Three age groups: 12 months old animals were sampled. Collected faeces were examined by quantitative flotation and Ziehl-Neelsen contrast staining, and species examined morphologically. Selected samples containing Cryptosporidium were additionally examined by polymerase-chain-reaction (PCR) and sequencing to determine genotypes. Twelve species of Eimeria were identified, seven previously unknown in Estonia. Main species in samples were E. bovis (30%), E. zuernii (23%), and E. ellipsoidalis (14%). All herds were infected and animals aged 3-12 months were more commonly infected with Eimeria oocysts (63%) than any other group. Calves Eimeria and the age category. Cryptosporidium were detected in 84% of the farms, and C. andersoni and C. parvum were successfully identified. Though prevalences of Cryptosporidium in the age groups were similar to the sample prevalence (30%) an increase in the infections was found with increasing age (pEimeria spp. infection (slope=-0.08, pEimeria species.

  19. Detection of Cryptosporidium sp infection by PCR and modified acid fast staining from potassium dichromate preserved stool

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    Agnes Kurniawan

    2009-09-01

    Full Text Available Aim To identify the frequency of Cryptosporidium infection in children below 3 years old by examining concentrated long term preserved stool using PCR detection of 18S rRNA gene and compared with modified acid fast staining technique.Methods Hundred eighty eight stools from children ≤ 3 years old were stored for 13 months in 2.5% K2Cr2O7 solution at 40C. Cryptosporidium oocysts were isolated by water-ether concentration technique. The concentrates were smeared onto object glass and stained with modified acid fast staining, and the rest of the concentrates were DNA extracted by freezing and thawing cycles and proteinase K digestion, then direct PCR was done to detect 18S rRNA gene.Result The proportion of positive stools for Cryptosporidium sp by acid fast staining from concentrated stools and 18S rRNA PCR were 4.8% and 34.6% respectively, which showed statistically significant difference.Conclusion The frequency of Cryptosporidium infection among children ≤ 3 years old was very high and stool storage in K2Cr2O7 for 13 months did not affect the PCR result. High prevalence of Cryptosporidium infection indicated high transmission in that area and the potential to be transmitted to other individuals such as the immunocompromised. (Med J Indones 2009;18:147-52Key words: 18S rRNA, cryptosporidiosis

  20. Clinical, serological, and parasitological analysis of snakes naturally infected with Cryptosporidium serpentis.

    Science.gov (United States)

    Paiva, Philipp Ricardo S O; Grego, Kathleen F; Lima, Valéria M F; Nakamura, Alex A; da Silva, Deuvânia C; Meireles, Marcelo V

    2013-11-15

    Infection by Cryptosporidium serpentis is one of the most important diseases in reptiles and is characterized by chronic clinical or subclinical infection and the presence of hypertrophic gastritis, food regurgitation, progressive weight loss, mortality, and intermittent or continuous shedding of oocysts in the feces. The objectives of this study were to standardize an indirect enzyme-linked immunosorbent assay (ELISA) to detect antibodies against C. serpentis and to evaluate the clinical, parasitological, and humoral immune response in snakes naturally infected with C. serpentis. Twenty-one snakes naturally infected with C. serpentis and housed at the Butantan Institute, São Paulo, Brazil, underwent clinical and parasitological analyses for C. serpentis infection through daily records of clinical signs and a monthly survey of fecal shedding of oocysts using the Kinyoun's acid-fast staining. The serological evaluation was performed monthly by indirect ELISA using crude total antigen from oocysts of C. serpentis to detect anti-C. serpentis antibodies. Clinical symptoms consisted of food regurgitation, inappetence, and progressive weight loss. The parasitological analysis revealed intermittent fecal shedding of a variable number of oocysts in all snakes, with positivity in 85.32% (157/184) of the samples. The indirect ELISA was positive in 68.25% (86/126) of the samples. A humoral immune response was observed in most animals; however, fluctuating antibodies levels, leading to alternating positive and negative results, were observed in most snakes.

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

    Science.gov (United States)

    Adamska, Małgorzata

    2015-02-01

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

  2. Small rodents as reservoirs of Cryptosporidium spp. and Giardia spp. in south-western Poland

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    Agnieszka Perec-Matysiak

    2015-02-01

    Full Text Available [i]Cryptosporidium[/i] spp. and [i]Giardia[/i] spp. have been detected in a range of host species, including rodents. The aim of this study was to determine the distribution of these pathogens and recognition of the reservoir role of rodents in the maintenance of these pathogens in south-western Poland. Additionally, preliminary molecular studies were conducted to elucidate the species and genotypes of [i]Cryptosporidium[/i] and [i]Giardia[/i] identified in this study. Stool samples (n=266 from [i]A. agrarius[/i],[i] A. flavicollis[/i] and [i]M. glareolus[/i], were subjected for analyses. Values of prevalence were 61.7, 68.3 and 68.1%, respectively, for [i]Cryptosporidium[/i] spp. and 41.7, 24.4 and 38.4%, respectively, for [i]Giardia[/i] spp. There was a statistically significant correlation between host species and [i]Giardia[/i] infection where[i] A. agrarius[/i] was the species of the highest prevalence. Statistically significant differences were not found for comparisons made for study sites and occurrence of [i]Giardia[/i] spp. and [i]Cryptosporidium[/i] spp. Due to preliminary nested PCR results, specific amplifications of [i]Cryptosporidium[/i] COWP and SSU rRNA genes were obtained for several isolates taken from rodent host species. One isolate recovered from [i]A. agrarius[/i] (from a semi-aquatic, urban area was identified as [i]C. parvum[/i] and revealed 100% similarity with sequences obtained from humans. To the best of the knowledge of the authors, this is the first record of the [i]C. parvum[/i] zoonotic species from the striped field mouse. Also recorded were the first findings of [i]C. ubiquitum[/i] from three small rodent species.

  3. MALDI-MIS INVESTIGATIONS OF DRINKING WATER PATHOGENS--GIARDIA AND CRYPTOSPORIDIUM

    Science.gov (United States)

    The protozoan parasites, Cryptosporidium parvum and Giardia lamblia, have been responsible for numerous waterborne outbreaks of gastrointestinal illness in the United States. The 1993 cryptosporidiosis outbreak in Milwaukee affected approximately 400,000 people and resulted in o...

  4. Literature Reference for Cryptosporidium spp. (Applied and Environmental Microbiology. 1999. 65(9): 3936–3941)

    Science.gov (United States)

    Procedures are described for analysis of animal samples using tissue culture techniques that may be adapted for assessment of solid, particulate, liquid and water samples contaminated with Cryptosporidium parvum.

  5. SPECIES AND GENUS DIFFERENTIATION OF PARASITES (GIARDIA AND CRYPTOSPORIDIUM) BY MALDI - MASS SPECTROMETRY

    Science.gov (United States)

    The protozoan parasites, Cryptosporidium parvum and Giardia lamblia, have been responsible for numerous waterborne outbreaks of gastrointestinal illness in the United States. The 1993 cryptosporidiosis outbreak in Milwaukee affected approximately 400,000 people and resulted in o...

  6. MALDI-MIS INVESTIGATIONS OF DRINKING WATER PATHOGENS--GIARDIA AND CRYPTOSPORIDIUM

    Science.gov (United States)

    The protozoan parasites, Cryptosporidium parvum and Giardia lamblia, have been responsible for numerous waterborne outbreaks of gastrointestinal illness in the United States. The 1993 cryptosporidiosis outbreak in Milwaukee affected approximately 400,000 people and resulted in o...

  7. Cryptosporidium spp. infection in mares and foals of the northwest region of São Paulo State, Brazil.

    Science.gov (United States)

    Inácio, Sandra Valéria; de Brito, Roberta Lomonte Lemos; Zucatto, Anaiza Simão; Coelho, Willian Marinho Dourado; de Aquino, Monally Conceição Costa; Aguirre, André de Abreu Rangel; Perri, Silvia Helena Venturoli; Meireles, Marcelo Vasconcelos; Bresciani, Katia Denise Saraiva

    2012-01-01

    The present study aimed to analyze the occurrence of infection by Cryptosporidium spp. in mares and their respective foals. This study was carried out in 11 farms located in the municipalities of Araçatuba, Birigui, Guararapes and Santo Antônio do Aracangua, in the northwest region of the State of Sao Paulo, from November 2010 to March 2011. A total of 98 mares and 98 foals of several breeds were analyzed; among foals, 59 were males and 39 females, aged from three to 330 days. Feces were collected directly from the rectal ampulla, purified and processed according to modified Kinyoun stain. Occurrence of Cryptosporidium spp. was 21.4% (21/98) for foals and 18.4% (18/98) for mares. Occurrence of Cryptosporidium spp. had significant association with breeds and age of animals. Results obtained led to the conclusion that foals older than two months and Mangalarga animals are less susceptible to the occurrence of Cryptosporidium spp.

  8. Cryptosporidium and Giardia in Danish organic pig farms: seasonal and age-related variation in prevalence, infection intensity and species/genotypes

    DEFF Research Database (Denmark)

    Petersen, Heidi Huus; Jianmin, Wang; Katakam, Kiran K.

    2015-01-01

    intensity was age-related for both parasites, and dual-infected pigs tended to excrete lower levels of oocysts compared to pigs harbouring only Cryptosporidium. Likewise, pigs infected with C. scrofarum excreted fewer oocysts (mean CPG: 54,848±194,508CI: 9085–118,781) compared......Although pigs are commonly infected with Cryptosporidium spp. and Giardia duodenalis, including potentially zoonotic species or genotypes, little is known about age-related infection levels, seasonal differences and genetic variation in naturally infected pigs raised in organic management systems....... Therefore, the current study was conducted to assess seasonal and age-related variations in prevalence and infection intensity of Cryptosporidium and Giardia, evaluate zoonotic potential and uncover correlations between species/genotypes, infection intensity and faecal consistency. Shedding of oocysts...

  9. Molecular characterization of Cryptosporidium isolates from beef calves under one month of age over three successive years in one herd in western France.

    Science.gov (United States)

    Rieux, Anaïs; Paraud, Carine; Pors, Isabelle; Chartier, Christophe

    2014-05-28

    Cohorts of pre-weaned calves were studied for Cryptosporidium infection over three successive years (2010-2012) in one beef cattle herd in western France. Each year 25-34 calves were sampled weekly from 3 days to one month of age in order to characterize oocyst output, Cryptosporidium species and clinical features associated with infection. Faecal samples were screened for the presence of oocysts using immunofluorescence analysis. DNA was extracted from positive samples and a PCR SSU rRNA followed by RFLP or sequencing was performed. For the subtyping of C. parvum, a gp60 PCR was carried out. Regardless of the year, 92-100% of the animals excreted oocysts on at least one sampling date. Depending on the year of observation, the age of highest prevalence varied. In contrast, the peak of excretion was systematically observed almost at the same age (2nd-3rd week of life) with excretion levels ranging from between 100 and 1.7 × 10(7)oocysts/g of faeces. Differences concerning clinical signs depending on the year of sampling were observed. Different species patterns were observed, with a predominance of C. bovis in the 1st year and a predominance of C. parvum in the last year. Moreover, two zoonotic subtypes of C. parvum, IIaA15G2R1 and IIaA18G2R1, were recorded in different years. This study shows that, in a given farm, the Cryptosporidium species and C. parvum subtypes identified as well as the prevalence of infection and level of excretion may vary greatly and show distinct patterns according to the year. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Pathological features of Cryptosporidium andersoni-induced lesions in SCID mice.

    Science.gov (United States)

    Masuno, Koichi; Yanai, Tokuma; Sakai, Hiroki; Satoh, Masaaki; Kai, Chieko; Nakai, Yutaka

    2013-07-01

    To assess the infectivity and the istopathological features of Cryptosporidium andersoni (C. andersoni) in laboratory animals, SCID mice were orally inoculated with oocysts of C. andersoni. Starting one week after inoculation, the SCID mice began shedding oocysts, and this continued for ten weeks. Histopathologically, myriads of C. andersoni were observed on the apical surface of the epithelium in the gastric pit of the glandular stomach. There were few lesions in the gastric epithelium except C. andersoni adhesion. In the lamina propria of the affected mucosa, minimum infiltration of inflammatory cells was observed. Immunohistochemically, C. andersoni demonstrated a positive reaction to a number of primary antibodies of Cryptosporidium parvum. In the experiment described here, few increases were seen in apoptotic epithelial cells in the affected mucosas of the SCID mice, and the nuclear augmentation was not enhanced. It was hypothesized that the absence of apoptosis and cell division were due to a lack of inflammatory cell reaction in the lamina propria.

  11. Molecular and phylogenetic characterization of Cryptosporidium and Giardia from pigs and cattle in Denmark

    DEFF Research Database (Denmark)

    Langkjær, Rikke Breinhold; Vigre, Håkan; Enemark, Heidi L.

    2007-01-01

    . suis, pig genotype II, C. parvum (cattle genotype), C. bovis, Cryptosporidium deer-like genotype and a novel C. suis-like genotype. For both cattle and pigs, a host age-related change in distribution of species/genotypes was observed. The zoonotic C. parvum (cattle genotype) was most prevalent in young...... purely by isolates of the livestock group, Assemblage E....

  12. AGING OF CRYPTOSPORIDIUM PARVUUM OOCYSTS STUDIED BY MALDI-TOF MS

    Science.gov (United States)

    Cryptosporidium parvum is a protozoan parasite, and it causes a potentially fatal gastrointestinal illness. This water borne pathogen has been the subject of several high profile disease outbreaks in the US and abroad. C. parvum presents challenges for both compliance monitorin...

  13. Identification of Cryptosporidium species and genotypes in dairy cattle in Brazil Identificação de espécies e genótipos de Cryptosporidium em bovinos leiteiros no Brasil

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    Flavio Medeiros Paz e Silva

    Full Text Available In this study, we identified Cryptosporidium species and genotypes present in dairy cattle in the central region of São Paulo state, Brazil. Fecal specimens were collected from 200 animals (100 calves and 100 cows in ten dairy farms. Fecal samples were examined using microscopic examination (ME, enzyme immunoassay (EIA and polymerase chain reaction (PCR. Cryptosporidium species and genotypes were determined by restriction fragment length polymorphism (RFLP or DNA sequencing analysis of the SSU-rRNA and GP60 genes. The occurrence of Cryptosporidium spp. infection was 14% (28/200. The occurrence in calves (26% was significantly higher than in cows (2%. Of the 27 Cryptosporidium-positive specimens submitted to genotyping, C. andersoni was identified in 23 (85.1%, C. bovis in three (11.1%, and the zoonotic C. parvum subtype IIaA15G2R1 in one (3.7%. The study demonstrates that Cryptosporidium spp. infection was common and widespread in dairy cattle in this region and that calves have a high prevalence of C. andersoni. Furthermore, the presence of C. parvum subtype IIaA15G2R1 indicates that dairy calves from this region should be considered a potential source of zoonotic Cryptosporidium oocysts.No presente estudo foram identificadas espécies e genótipos de Cryptosporidium originadas de bovinos leiteiros na região central do estado de São Paulo, Brasil. Amostras fecais foram coletadas de 200 animais (100 bezerros e 100 vacas em 10 propriedades leiteiras. As amostras foram examinadas utilizando os métodos de microscopia óptica (MO, ensaio imunoenzimático (EI e reação em cadeia da polimerase (PCR. As espécies e genótipos de Cryptosporidium foram determinados pelo método de polimorfismo no tamanho dos fragmentos de restrição (RFLP ou sequenciamento dos genes SSU-rRNA e GP60. A infecção por Cryptosporidium spp. teve ocorrência de 14% (28/200. A ocorrência em bezerros (26% foi significativamente maior do que em vacas (2%. Do total de 27

  14. Genetic diversity of Cryptosporidium identified in clinical samples from cities in Brazil and Argentina

    Science.gov (United States)

    Peralta, Regina Helena Saramago; Velásquez, Jorge Néstor; Cunha, Flavia de Souza; Pantano, María Laura; Sodré, Fernando Campos; da Silva, Sidnei; Astudillo, Osvaldo Germán; Peralta, José Mauro; Carnevale, Silvana

    2016-01-01

    The identification and characterisation of Cryptosporidiumgenotypes and subtypes are fundamental to the study of cryptosporidiosis epidemiology, aiding in prevention and control strategies. The objective was to determine the genetic diversity ofCryptosporidium in samples obtained from hospitals of Rio de Janeiro, Brazil, and Buenos Aires, Argentina. Samples were analysed by microscopy and TaqMan polymerase chain reaction (PCR) assays forCryptosporidium detection, genotyped by nested-PCR-restriction fragment length polymorphism (RFLP) analysis of the 18S rRNA gene and subtyped by DNA sequencing of the gp60 gene. Among the 89 samples from Rio de Janeiro, Cryptosporidium spp were detected in 26 by microscopy/TaqMan PCR. In samples from Buenos Aires,Cryptosporidium was diagnosed in 15 patients of the 132 studied. The TaqMan PCR and the nested-PCR-RFLP detected Cryptosporidium parvum, Cryptosporidium hominis, and co-infections of both species. In Brazilian samples, the subtypes IbA10G2 and IIcA5G3 were observed. The subtypes found in Argentinean samples were IbA10G2, IaA10G1R4, IaA11G1R4, and IeA11G3T3, and mixed subtypes of Ia and IIa families were detected in the co-infections. C. hominis was the species more frequently detected, and subtype family Ib was reported in both countries. Subtype diversity was higher in Buenos Aires than in Rio de Janeiro and two new subtypes were described for the first time. PMID:26814641

  15. Genetic diversity of Cryptosporidium identified in clinical samples from cities in Brazil and Argentina

    Directory of Open Access Journals (Sweden)

    Regina Helena Saramago Peralta

    2016-01-01

    Full Text Available The identification and characterisation of Cryptosporidiumgenotypes and subtypes are fundamental to the study of cryptosporidiosis epidemiology, aiding in prevention and control strategies. The objective was to determine the genetic diversity ofCryptosporidium in samples obtained from hospitals of Rio de Janeiro, Brazil, and Buenos Aires, Argentina. Samples were analysed by microscopy and TaqMan polymerase chain reaction (PCR assays forCryptosporidium detection, genotyped by nested-PCR-restriction fragment length polymorphism (RFLP analysis of the 18S rRNA gene and subtyped by DNA sequencing of the gp60 gene. Among the 89 samples from Rio de Janeiro, Cryptosporidium spp were detected in 26 by microscopy/TaqMan PCR. In samples from Buenos Aires,Cryptosporidium was diagnosed in 15 patients of the 132 studied. The TaqMan PCR and the nested-PCR-RFLP detected Cryptosporidium parvum, Cryptosporidium hominis, and co-infections of both species. In Brazilian samples, the subtypes IbA10G2 and IIcA5G3 were observed. The subtypes found in Argentinean samples were IbA10G2, IaA10G1R4, IaA11G1R4, and IeA11G3T3, and mixed subtypes of Ia and IIa families were detected in the co-infections. C. hominis was the species more frequently detected, and subtype family Ib was reported in both countries. Subtype diversity was higher in Buenos Aires than in Rio de Janeiro and two new subtypes were described for the first time.

  16. The role of the multiple banded antigen of Ureaplasma parvum in intra-amniotic infection: major virulence factor or decoy?

    Science.gov (United States)

    Dando, Samantha J; Nitsos, Ilias; Kallapur, Suhas G; Newnham, John P; Polglase, Graeme R; Pillow, J Jane; Jobe, Alan H; Timms, Peter; Knox, Christine L

    2012-01-01

    The multiple banded antigen (MBA) is a predicted virulence factor of Ureaplasma species. Antigenic variation of the MBA is a potential mechanism by which ureaplasmas avoid immune recognition and cause chronic infections of the upper genital tract of pregnant women. We tested whether the MBA is involved in the pathogenesis of intra-amniotic infection and chorioamnionitis by injecting virulent or avirulent-derived ureaplasma clones (expressing single MBA variants) into the amniotic fluid of pregnant sheep. At 55 days of gestation pregnant ewes (n = 20) received intra-amniotic injections of virulent-derived or avirulent-derived U. parvum serovar 6 strains (2×10⁴ CFU), or 10B medium (n = 5). Amniotic fluid was collected every two weeks post-infection and fetal tissues were collected at the time of surgical delivery of the fetus (140 days of gestation). Whilst chronic colonisation was established in the amniotic fluid of animals infected with avirulent-derived and virulent-derived ureaplasmas, the severity of chorioamnionitis and fetal inflammation was not different between these groups (p>0.05). MBA size variants (32-170 kDa) were generated in vivo in amniotic fluid samples from both the avirulent and virulent groups, whereas in vitro antibody selection experiments led to the emergence of MBA-negative escape variants in both strains. Anti-ureaplasma IgG antibodies were detected in the maternal serum of animals from the avirulent (40%) and virulent (55%) groups, and these antibodies correlated with increased IL-1β, IL-6 and IL-8 expression in chorioamnion tissue (p<0.05). We demonstrate that ureaplasmas are capable of MBA phase variation in vitro; however, ureaplasmas undergo MBA size variation in vivo, to potentially prevent eradication by the immune response. Size variation of the MBA did not correlate with the severity of chorioamnionitis. Nonetheless, the correlation between a maternal humoral response and the expression of chorioamnion cytokines is a

  17. The role of the multiple banded antigen of Ureaplasma parvum in intra-amniotic infection: major virulence factor or decoy?

    Directory of Open Access Journals (Sweden)

    Samantha J Dando

    Full Text Available The multiple banded antigen (MBA is a predicted virulence factor of Ureaplasma species. Antigenic variation of the MBA is a potential mechanism by which ureaplasmas avoid immune recognition and cause chronic infections of the upper genital tract of pregnant women. We tested whether the MBA is involved in the pathogenesis of intra-amniotic infection and chorioamnionitis by injecting virulent or avirulent-derived ureaplasma clones (expressing single MBA variants into the amniotic fluid of pregnant sheep. At 55 days of gestation pregnant ewes (n = 20 received intra-amniotic injections of virulent-derived or avirulent-derived U. parvum serovar 6 strains (2×10⁴ CFU, or 10B medium (n = 5. Amniotic fluid was collected every two weeks post-infection and fetal tissues were collected at the time of surgical delivery of the fetus (140 days of gestation. Whilst chronic colonisation was established in the amniotic fluid of animals infected with avirulent-derived and virulent-derived ureaplasmas, the severity of chorioamnionitis and fetal inflammation was not different between these groups (p>0.05. MBA size variants (32-170 kDa were generated in vivo in amniotic fluid samples from both the avirulent and virulent groups, whereas in vitro antibody selection experiments led to the emergence of MBA-negative escape variants in both strains. Anti-ureaplasma IgG antibodies were detected in the maternal serum of animals from the avirulent (40% and virulent (55% groups, and these antibodies correlated with increased IL-1β, IL-6 and IL-8 expression in chorioamnion tissue (p<0.05. We demonstrate that ureaplasmas are capable of MBA phase variation in vitro; however, ureaplasmas undergo MBA size variation in vivo, to potentially prevent eradication by the immune response. Size variation of the MBA did not correlate with the severity of chorioamnionitis. Nonetheless, the correlation between a maternal humoral response and the expression of chorioamnion

  18. Flowering as the Most Highly Sensitive Period of Grapevine (Vitis vinifera L. cv Mourvèdre to the Botryosphaeria Dieback Agents Neofusicoccum parvum and Diplodia seriata Infection

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    Alessandro Spagnolo

    2014-05-01

    Full Text Available Botryosphaeria dieback is a fungal grapevine trunk disease that currently represents a threat for viticulture worldwide because of the important economical losses due to reduced yield of affected plants and their premature death. Neofusicoccum parvum and Diplodia seriata are among the causal agents. Vine green stems were artificially infected with N. parvum or D. seriata at the onset of three different phenological stages (G stage (separated clusters, flowering and veraison. Highest mean lesion lengths were recorded at flowering. Major proteome changes associated to artificial infections during the three different phenological stages were also reported using two dimensional gel electrophoresis (2D-based analysis. Twenty (G stage, 15 (flowering and 13 (veraison differentially expressed protein spots were subjected to nanoLC-MS/MS and a total of 247, 54 and 25 proteins were respectively identified. At flowering, a weaker response to the infection was likely activated as compared to the other stages, and some defense-related proteins were even down regulated (e.g., superoxide dismutase, major latex-like protein, and pathogenesis related protein 10. Globally, the flowering period seemed to represent the period of highest sensitivity of grapevine to Botryosphaeria dieback agent infection, possibly being related to the high metabolic activity in the inflorescences.

  19. Flowering as the Most Highly Sensitive Period of Grapevine (Vitis vinifera L. cv Mourvèdre) to the Botryosphaeria Dieback Agents Neofusicoccum parvum and Diplodia seriata Infection

    Science.gov (United States)

    Spagnolo, Alessandro; Larignon, Philippe; Magnin-Robert, Maryline; Hovasse, Agnès; Cilindre, Clara; Van Dorsselaer, Alain; Clément, Christophe; Schaeffer-Reiss, Christine; Fontaine, Florence

    2014-01-01

    Botryosphaeria dieback is a fungal grapevine trunk disease that currently represents a threat for viticulture worldwide because of the important economical losses due to reduced yield of affected plants and their premature death. Neofusicoccum parvum and Diplodia seriata are among the causal agents. Vine green stems were artificially infected with N. parvum or D. seriata at the onset of three different phenological stages (G stage (separated clusters), flowering and veraison). Highest mean lesion lengths were recorded at flowering. Major proteome changes associated to artificial infections during the three different phenological stages were also reported using two dimensional gel electrophoresis (2D)-based analysis. Twenty (G stage), 15 (flowering) and 13 (veraison) differentially expressed protein spots were subjected to nanoLC-MS/MS and a total of 247, 54 and 25 proteins were respectively identified. At flowering, a weaker response to the infection was likely activated as compared to the other stages, and some defense-related proteins were even down regulated (e.g., superoxide dismutase, major latex-like protein, and pathogenesis related protein 10). Globally, the flowering period seemed to represent the period of highest sensitivity of grapevine to Botryosphaeria dieback agent infection, possibly being related to the high metabolic activity in the inflorescences. PMID:24886812

  20. Detection and molecular characterization of Giardia and Cryptosporidium in common dolphins (Delphinus delphis) stranded along the Galician coast (Northwest Spain).

    Science.gov (United States)

    Reboredo-Fernández, A; Gómez-Couso, H; Martínez-Cedeira, J A; Cacciò, S M; Ares-Mazás, E

    2014-05-28

    The ubiquitous protozoan parasites Giardia and Cryptosporidium have been detected from many species of captive and free-living wildlife, representing most mammalian orders. Twenty species of marine mammals have been reported to inhabit Galician waters and the region has one of the highest rates of stranding in Europe. Evidence from stranding, reported by-catches and sightings, suggests that the common dolphin (Delphinus delphis) is the most abundant cetacean on the Galician coast (Northwest Spain). The objective of this study was to detect and molecularly characterize isolates of Giardia and Cryptosporidium obtained from common dolphins stranded in this area. Between 2005 and 2012, sections of large intestine from 133 common dolphins stranded along the Galician coast were collected by the personnel of the Galician Stranding Network (Coordinadora para o Estudo dos Mamíferos Mariños, CEMMA). Using direct immunofluorescence antibody test (IFAT) and PCR amplification and sequencing of the SSU-rDNA, β-giardin genes and the ITS1-5.8S-ITS2 region, Giardia and Cryptosporidium were detected in 8 (6.0%) and 12 samples (9.0%), respectively. In two samples, co-infection by both parasites was observed. The molecular characterization revealed the presence of Giardia duodenalis assemblages A (genotypes A1 and A2) and B and Cryptosporidium parvum in these samples. This constitutes the first study in which the presence of Giardia and Cryptosporidium has been investigated in common dolphins on the European Atlantic coast, and it is also the first report of C. parvum in this host. Our findings indicate that these animals could act as reservoir of these waterborne parasites or could be victims of the contamination originated by anthropogenic activities.

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

    Science.gov (United States)

    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

    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.

  2. A Survey Study of Cryptosporidium Infection in Children under 10 Years Old Referred to the Health Care Centers of Hamadan District in 2013

    Directory of Open Access Journals (Sweden)

    M. Asadi

    2014-10-01

    Full Text Available Introduction & Objective: Cryptosporidium is one of the most important zoonotic and oppor-tunistic protozoa and can cause diarrhea in those with impaired immune systems, as well as the children. Considering the high sensitivity of children against infection caused by crypto-sporidium, its zoonotic nature and lack of treatment, this study aimed to determine the prevalence of cryptosporidium infection in children under 10 years old, referred to the health care centers of Hamadan district. Materials & Methods: This study was conducted in 2013 on 420 children (222 males and 198 females, who were referred to urban and rural health care centers in Hamadan district. Stool samples were examined using formalin-ether method and modified Ziehl-Neelsen staining technique. The results were analyzed with chi-square test. Results: Of the 420 children studied, 2 individuals (0.47% (A 16-month-old boy and a 6-year-old girl were infected with cryptosporidium spp. The infection was observed only in rural areas and in children that were in direct contact with the animals. Conclusion: The results of this study showed a presence of cryptosporidium in rural areas compared to urban areas and in children in contact with animals. Therefore it is necessary to promote the public health awareness of rural population. (Sci J Hamadan Univ Med Sci 2014; 21 (3: 211-217

  3. Neofusicoccum parvum Colonization of the Grapevine Woody Stem Triggers Asynchronous Host Responses at the Site of Infection and in the Leaves

    Directory of Open Access Journals (Sweden)

    Mélanie Massonnet

    2017-06-01

    Full Text Available Grapevine trunk diseases cause important economic losses in vineyards worldwide. Neofusicoccum parvum, one of the most aggressive causal agents of the trunk disease Botryosphaeria dieback, colonizes cells and tissues of the grapevine wood, leading to the formation of an internal canker. Symptoms then extend to distal shoots, with wilting of leaves and bud mortality. Our aim was to characterize the transcriptional dynamics of grapevine genes in the woody stem and in the leaves during Neofusicoccum parvum colonization. Genome-wide transcriptional profiling at seven distinct time points (0, 3, and 24 hours; 2, 6, 8, and 12 weeks showed that both stems and leaves undergo extensive transcriptomic reprogramming in response to infection of the stem. While most intense transcriptional responses were detected in the stems at 24 hours, strong responses were not detected in the leaves until the next sampling point at 2 weeks post-inoculation. Network co-expression analysis identified modules of co-expressed genes common to both organs and showed most of these genes were asynchronously modulated. The temporal shift between stem vs. leaf responses affected transcriptional modulation of genes involved in both signal perception and transduction, as well as downstream biological processes, including oxidative stress, cell wall rearrangement and cell death. Promoter analysis of the genes asynchronously modulated in stem and leaves during N. parvum colonization suggests that the temporal shift of transcriptional reprogramming between the two organs might be due to asynchronous co-regulation by common transcriptional regulators. Topology analysis of stem and leaf co-expression networks pointed to specific transcription factor-encoding genes, including WRKY and MYB, which may be associated with the observed transcriptional responses in the two organs.

  4. Molecular characterization of Cryptosporidium spp. in pre-weaned kids in a dairy goat farm in western France.

    Science.gov (United States)

    Rieux, Anais; Paraud, Carine; Pors, Isabelle; Chartier, Christophe

    2013-02-18

    A longitudinal study was undertaken to characterize the course of Cryptosporidium infection in a dairy goat farm located in western France. Two cohorts of twenty-five and fifteen animals, respectively, were sampled once a week from birth to weaning. Each individual fecal sample was screened using direct immunofluorescence (IFT) and if found positive, the Cryptosporidium species was identified using PCR analysis. Seventeen (68% [95% CI: 44-91]) animals were positive at least once during the first study and 14 (93% [95% CI: 80-100]) during the second, after IFT examination. In the first study, the age at first excretion was 17 days and the peak of excretion (mean arithmetic excretion: 22,700 oocysts per gram (opg) of feces) was recorded when kids were between 22 and 28 days old. For the second study, the age at first excretion was 10 days and the peak of excretion (mean arithmetic excretion: 3.4 × 10(6)opg) was recorded in animals aged between 10 and 14 days. Clinical signs were observed only in animals of the second cohort. DNA sequence analysis at the 18S ribosomal RNA locus was successful for 9 of the 27 IFT-positive samples in the first cohort and for 10 of the 34 positive isolates in the second cohort. All isolates were identified as Cryptosporidium xiaoi except one which was identified as Cryptosporidium parvum. Our results confirm that goat kids are hosts for C. parvum and C. xiaoi and that infection by C. xiaoi may be associated with mild clinical signs. Copyright © 2012 Elsevier B.V. All rights reserved.

  5. Cryptosporidium and Giardia in Danish organic pig farms: Seasonal and age-related variation in prevalence, infection intensity and species/genotypes.

    Science.gov (United States)

    Petersen, Heidi H; Jianmin, Wang; Katakam, Kiran K; Mejer, Helena; Thamsborg, Stig M; Dalsgaard, Anders; Olsen, Annette; Enemark, Heidi L

    2015-11-30

    Although pigs are commonly infected with Cryptosporidium spp. and Giardia duodenalis, including potentially zoonotic species or genotypes, little is known about age-related infection levels, seasonal differences and genetic variation in naturally infected pigs raised in organic management systems. Therefore, the current study was conducted to assess seasonal and age-related variations in prevalence and infection intensity of Cryptosporidium and Giardia, evaluate zoonotic potential and uncover correlations between species/genotypes, infection intensity and faecal consistency. Shedding of oocysts and cysts ((oo-)cysts) was monitored at quarterly intervals (September 2011-June 2012) in piglets (n = 152), starter pigs (n = 234), fatteners (n = 230) and sows (n = 240) from three organic farms in Denmark. (oo-)Cysts were quantified by immunofluorescence microscopy; and 56/75 subsamples from Cryptosporidium infected pigs were successfully analysed by PCR amplification and partial sequencing of the small subunit (SSU) 18S rRNA and hsp70genes, while 13/67 Giardia subsamples were successfully analysed by amplification and partial sequencing of the 18S rRNA and the gdh genes. Altogether, Cryptosporidium or Giardia infections were observed in 40.9% (350/856) and 14.0% (120/856) of the pigs, respectively, including 8.2% (70/856) infected with both parasites. Prevalence, intensity of infections and presence of Cryptosporidium species varied significantly between age-groups; 53.3% piglets, 72.2% starter pigs, 40.4% fatteners and 2.9% sows were infected with Cryptosporidium, whereas 2.0% piglets, 27.4% starter pigs, 17.8% fatteners and 5.0% sows were infected with Giardia. The overall prevalence was stable throughout the year, except for dual-infections that were more prevalent in September and December (p parasites, and dual-infected pigs tended to excrete lower levels of oocysts compared to pigs harbouring only Cryptosporidium. Likewise, pigs infected with Cryptosporidium

  6. Cryptosporidiosis in Haiti: surprisingly low level of species diversity revealed by molecular characterization of Cryptosporidium oocysts from surface water and groundwater

    Science.gov (United States)

    Damiani, Céline; Balthazard-Accou, Ketty; Clervil, Elmyre; Diallo, Aïssata; Da Costa, Cécilia; Emmanuel, Evens; Totet, Anne; Agnamey, Patrice

    2013-01-01

    The protozoan parasite Cryptosporidium sp. has emerged as one of the most important water contaminants, causing waterborne outbreaks of diarrhoeal diseases worldwide. In Haiti, cryptosporidiosis is a frequent cause of diarrhoea in children under the age of five years, HIV-infected individuals, and people living in low socioeconomic conditions, mainly due to the consumption of water or food polluted by Cryptosporidium oocysts. The aim of this study was to detect and identify Cryptosporidium oocysts present in 12 water samples collected in Port-au-Prince and 4 water samples collected in Cap Haïtien. Initial detection consisted of immunomagnetic separation – immunofluorescence assay (IMS-IFA), which was confirmed by nested PCR, targeting the most polymorphic region of the 18S rRNA gene in 15/16 samples. Genotyping was performed by PCR-restriction fragment length polymorphism (RFLP) analysis and DNA sequencing. Under our working conditions, neither nested PCR-RFLP nor direct DNA sequencing revealed the expected species diversity, as only Cryptosporidium parvum was identified in the water samples studied. This study highlights the difficulty of detecting mixed populations of Cryptosporidium species in environmental samples. PMID:24252814

  7. Cryptosporidium and Giardia: new challenges to the water industry

    NARCIS (Netherlands)

    Medema, Gerriet Jan

    2001-01-01

    The protozoan parasites Cryptosporidium parvum and Giardia intestinalis have emerged as significant waterborne pathogens over the past decades. Many outbreaks of waterborne cryptosporidiosis and giardiasis have been recorded,primarily in the United States and the United Kingdom.Chapter 1 gives an ov

  8. A study of risk factors associated with the prevalence of Cryptosporidium in villages around Lake Atitlan, Guatemala

    Directory of Open Access Journals (Sweden)

    H. E. Laubach

    2004-08-01

    Full Text Available Cryptosporidium parvum is an endemic, zoonotic coccidian parasitosis that is highly prevalent in third-world countries where waterborne fecal contamination of food and drink or person-to-person contact with oocysts are the most common methods of transmission of the enteric protozoan. This type of transmission of the parasite made the villages around Lake Atitlan, Guatemala a unique site to compare environmental risk factors with the level of Cryptosporidium infections in the local residents. The study was carried out in two villages, San Antonio Palopo and Santa Catarina Palopo, located in the highlands above the shores of the lake. Smears from stool specimens of patients with gastroenteritis were processed using Kinyoun's modified acid-fast stain and observed with light microscopy. Of the 100 residents examined from the two villages, 32% had Cryptosporidium infections. Female children had the highest prevalence of infection (44% in San Antonio Palopo and 46% in Santa Catarina Palopo, p<0.05, and they also had significantly higher infection rates than males, 50% vs. 17%, respectively. The prevalence rate was not influenced by the season of the year or by the location of the residents. We found differences in prevalence rates due to age and gender, and we suggest that the high infection rates of specific groups are associated with their exposure to the contaminated water supply from Lake Atitlan.

  9. Cryptosporidium and Giardia in Danish organic pig farms: seasonal and age-related variation in prevalence, infection intensity and species/genotypes

    DEFF Research Database (Denmark)

    Petersen, Heidi Huus; Jianmin, Wang; Katakam, Kiran K.;

    2015-01-01

    infected pigs were successfully analysed by PCR amplification and partial sequencing of the small subunit (SSU) 18S rRNA and hsp70genes, while 13/67 Giardia subsamples were successfully analysed by amplification and partial sequencing of the 18S rRNA and the gdh genes. Altogether, Cryptosporidium...

  10. Assessing calves as carriers of Cryptosporidium and Giardia with zoonotic potential on dairy and beef farms within a water catchment area by mutation scanning.

    Science.gov (United States)

    Abeywardena, Harshanie; Jex, Aaron R; Firestone, Simon M; McPhee, Sandra; Driessen, Nicole; Koehler, Anson V; Haydon, Shane R; von Samson-Himmelstjerna, Georg; Stevens, Melita A; Gasser, Robin B

    2013-08-01

    In the present study, we undertook a molecular epidemiological survey of Cryptosporidium and Giardia in calves on three dairy and two beef farms within an open drinking water catchment area (Melbourne, Australia). Faecal samples (n = 474) were collected from calves at two time points (5 months apart) and tested using a PCR-based mutation scanning-targeted sequencing phylogenetic approach, employing regions within the genes of small subunit (SSU) of ribosomal RNA (designated partial SSU), 60 kDa glycoprotein (pgp60) and triose phosphate isomerase (ptpi) as genetic markers. Using partial SSU, the C. bovis, C. parvum, C. ryanae and a new genotype of Cryptosporidium were characterised from totals of 74 (15.6%), 35 (7.3%), 37 (7.8%) and 9 (1.9%) samples, respectively. Using pgp60, C. parvum genotype IIa subgenotype A18G3R1 was detected in 29 samples. Using ptpi, G. duodenalis assemblages A and E were detected in totals of 10 (2.1%) and 130 (27.4%) samples, respectively. The present study showed that a considerable proportion of dairy and beef calves in this open water catchment region excreted Cryptosporidium (i.e. subgenotype IIaA18G3R1) and Giardia (e.g. assemblage A) that are consistent with those infecting humans, inferring that they are of zoonotic importance. Future work should focus on exploring, in a temporal and spatial way, whether these parasites occur in the environment and water of the catchment reservoir.

  11. Comparação entre os métodos de Ziehl-Neelsen modificado e Acid-Fast-Trichrome para a pesquisa fecal de Cryptosporidium parvum e Isospora belli

    OpenAIRE

    Rigo,Carla R.; Franco,Regina Maura B.

    2002-01-01

    Devido a crescente importância dos coccídios intestinais (Cryptosporidium, Isospora e Cyclospora) como parasitos oportunistas, é fundamental para os laboratórios diferenciar morfologicamente estes protozoários; a técnica de Ziehl-Neelsen modificada (ZNm) é amplamente utilizada para este fim; recentemente, foi proposto um novo procedimento, a coloração combinada do ácido tricrômico (Acid-Fast-Trichrome - AFT). O objetivo do presente estudo foi comparar os processos AFT e ZNm para a detecção de...

  12. Cryptosporidium: from laboratory diagnosis to surveillance and outbreaks

    Directory of Open Access Journals (Sweden)

    Chalmers R.M.

    2008-09-01

    Full Text Available The burden of disease caused by the protozoan parasite Cryptosporidium is unknown. However, routine laboratory diagnosis and surveillance enables the basic epidemiology to be described, changes to be monitored and under-ascertainment to be measured. Although the two main species involved in human disease in developed countries, Cryptosporidium parvum and Cryptosporidium hominis, have differing epidemiologies and risk factors, national surveillance is generally from isolates identified to the genus level only. Enhancing the data by typing, at least to identify the isolates to the species level, removes some of the noise generated and better identifies the risks than when reports are not species-specific. This level of identification is also valuable for outbreak investigations, but further investigation of the population genetics of C. parvum and C. hominis is required for the development of more readily applied subtyping tools.

  13. Cryptosporidium, Enterocytozoon, and Cyclospora Infections in Pediatric and Adult Patients with Diarrhea in Tanzania.

    OpenAIRE

    Cegielski, J P; Ortega, Y. R.; McKee, S; Madden, J F; Gaido, L; Schwartz, D. A.; Manji, K; Jorgensen, A F; Miller, S E; Pulipaka, U P; Msengi, A E; Mwakyusa, D H; Sterling, C R; Reller, L B

    1999-01-01

    Cryptosporidiosis, microsporidiosis, and cyclosporiasis were studied in four groups of Tanzanian inpatients: adults with AIDS-associated diarrhea, children with chronic diarrhea (of whom 23 of 59 were positive [+] for human immunodeficiency virus [HIV]), children with acute diarrhea (of whom 15 of 55 were HIV+), and HIV control children without diarrhea. Cryptosporidium was identified in specimens from 6/86 adults, 5/59 children with chronic diarrhea (3/5, HIV+), 7/55 children with acute diar...

  14. Genetic characterisation of Cryptosporidium and Giardia from dairy calves: discovery of species/genotypes consistent with those found in humans.

    Science.gov (United States)

    Abeywardena, Harshanie; Jex, Aaron R; Nolan, Matthew J; Haydon, Shane R; Stevens, Melita A; McAnulty, Robin W; Gasser, Robin B

    2012-12-01

    Cryptosporidium and Giardia are important genera of parasitic protists that can cause significant diarrhoeal diseases in humans and other animals. Depending on the species/genotype of parasite, human infection may be acquired via anthroponotic or zoonotic transmission routes. Here, we undertook a molecular epidemiological investigation of these two genera of parasites in pre- and post-weaned calves from eight locations in Canterbury, New Zealand, by PCR-coupled sequencing and phylogenetic analysis of sequence data for regions in the 60 kDa glycoprotein (pgp60) gene of Cryptosporidium and/or the triose-phosphate isomerase (ptpi) gene of Giardia. The pgp60 and ptpi regions were specifically amplified from 15 (8.3%) and 11 (6.1%) of the 180 individual faecal samples, respectively. The sequences derived from all of the amplicons were aligned with homologous reference sequences and subjected to phylogenetic analysis by Bayesian inference. For Cryptosporidium, three samples contained Cryptosporidium parvum genotype IIa, subgenotypes IIaA15G3R1, IIaA19G3R1 and IIaA23G4. Twelve samples contained Cryptosporidium hominis genotype Ib, subgenotype IbA10G2R2. While subgenotypes IIaA15G3R1 and IIaA23G4 are new records, IIaA19G3R1 and IbA10G2R2 are commonly found in humans in various countries. For Giardia, two samples contained Giardia duodenalis assemblage A, also common in humans. In contrast, nine samples contained G. duodenalis assemblage E, which is the first report of this assemblage in cattle in New Zealand. Therefore, the present results indicate that dairy calves on the South Island of New Zealand harbour 'zoonotic' genotypes of Cryptosporidium and Giardia, which is likely to have significant public health implications.

  15. DETECTION OF CRYPTOSPORIDIUM OOCYSTS AND SERUM IMMUNOGLOBULIN G (LGG ANTIBODIES IN NATURALLY INFECTED CALVES

    Directory of Open Access Journals (Sweden)

    Rahmatullah Rind, A.J. Probert1 and M.I. Rind2

    2000-01-01

    Full Text Available Sixty three faecal as well as blood samples from a group of 15 young Friesian calves under 2 months of age at Aber Farm Bangor, U.K. were collected on monthly basis and examined for the presence of Cryptosporidium oocysts and serum immunoglobulin G (IgG antibodies, Twelve (19.23 % were found positive with Cryptosporium species while in 5 (7.9 % faecal samples both Cryptosporidium and Eimeria were present but 46 (73.0 % samples were negative. In 9 out of 12 (75.0 % cases where Cryptosporidium ocysts were present, a positive IF AT was observed while in 4 out of 5 (80.0 % positives were seen in the presence of both Cryptosporium and Eimeria oocysts. In contrast only 6 out of 46 (13.1% cases, a positive IFAT was also seen when no oocysts were recorded. Oocysts fluoresced brightly with positive serum samples and only faintly or not at all with the negative samples or the conjugate alone.

  16. [Cryptosporidium hominis diarrhea outbreak and transmission linked to diaper infant use].

    Science.gov (United States)

    Teresa Ortega, María; Vergara, Alberto; Guimbao, Joaquín; Clavel, Antonio; Gavín, Patricia; Ruiz, Andrés

    2006-11-04

    On the basis of several cases of cryptosporidiosis detected in a child day-care center, we stablished the extent of the outbreak and investigated causes of parasite transmission. A retrospective cohort study was designed on all children attending day-care center and care givers to determine their infection status and identify risk factors associated to the outbreak. 24 cases of cryptosporidiosis were detected, with an attack rate of 0.46 (24/52); 12 of them were parasitologycally confirmed. All care givers were negative for Cryptosporidium and none of them reported symptoms of acute gastroenteritis. Transmission pattern was compatible with person to person modes. Among the factors investigated, two were associated with the risk of disease: diaper wear (relative risk = 2.06; p = 0.059); and diarrhea in relatives (relative risk = 2.05; p = 0.01). In all confirmed cases, Cryptosporidium hominis (previously known as C. parvum, genotype 1), was identified. Cryptosporidiosis should be considered as a possible cause of outbreaks of gastroenteritis at day-care centers. Increasing care on diaper changing practices, specially over children with diarrhea, may be the key factor to prevent transmission of Cryptosporidium.

  17. Effective Concentration and Detection of Cryptosporidium, Giardia, and the Microsporidia from Environmental Matrices

    Directory of Open Access Journals (Sweden)

    Joseph A. Moss

    2014-01-01

    Full Text Available Cryptosporidium spp., Giardia spp., and members of Microsporidia are enteropathogenic parasites of humans and animals, producing asymptomatic to severe intestinal infections. To circumvent various impediments associated with current detection methods, we tested a method providing multistage purification and separation in a single, confined step. Standard real-time PCR was used as a detection method. Samples spiked with C. parvum and G. intestinalis were split for comparison to standard Method 1623. Results were equivalent to immunomagnetic procedures for Cryptosporidium, and Giardia. Overall percent recovery for Cryptosporidium with Method 1623 averaged 26.89% (std 21.44%; min = 0%; max = 73% and was similar but less variable for qPCR method at an estimated average of 27.67 (std 17.65%; min = 5%; max = 63%. For Giardia, Method 1623 had an overall average recovery of 27.11% (std 17.98%; min = 1%; max = 58%, while multistage purification and qPCR had an estimated lower overall recovery at 18.58% (std 13.95%; min = 0%; max = 35%. Microsporidia were also readily detected with an estimated recovery of 46.81% overall (std 17.66%; min = 18%; max = 70% for E. intestinalis and 38.90% (std 14.36%; min = 13%; max = 62% for E. bieneusi.

  18. Infecção natural por Cryptosporidium sp. em aves domésticas comercializadas em mercados municipais do Estado do Rio de Janeiro Natural infection by Cryptosporidium sp. in domestic fowls sold at local markets

    Directory of Open Access Journals (Sweden)

    Raquel Saucier Gomes

    2009-10-01

    Full Text Available O objetivo do trabalho foi comparar a dinâmica, a ocorrência, a morfometria de oocistos e os períodos patentes de Cryptosporidium sp. em aves domésticas, patos (Anas platyrhynchos, pintos (Gallus gallus e codornas (Coturnix japonica, naturalmente infectadas, provenientes de dois mercados municipais do Estado do Rio de Janeiro, Rio de Janeiro (RJ. Houve diferenças quanto à ocorrência da infecção entre os dois locais e entre pintos e patos, mas não entre codornas. Para a morfometria, foram observadas diferenças estatísticas nas medidas dos diâmetros maior e menor e para o índice morfométrico calculado (P0,05. Na comparação do período de eliminação, patos tiveram um maior período com maiores quantidades de oocistos eliminados. Codornas e pintos apresentaram dinâmica de eliminação semelhante e não houve diferença quanto à concentração de oocistos. Pintos foram mais susceptíveis à infecção seguidos por patos e codornas. Pode-se concluir que a infecção natural por Cryptoporidium sp. foi frequente nas aves estudadas. Patos, pintos e codornas podem ser disseminadores do protozoário em mercados municipais do Rio de Janeiro, RJ. Assim, podem constituir risco de infecção.The objective of the current study was comparing the dynamic and occurrence of Cryptosporidium sp., as well as the morphometry and elimination period of oocysts in naturally infected ducks (Anas platyrhynchos, chickens (Gallus gallus and Japanese quails (Coturnix japonica from two local markets of Rio de Janeiro, RJ. There were significant differences considering the occurrence of infection between the two markets, and also between chickens and ducks, but not among Japanese quails. Also, significant statistical differences were observed in morphometry, considering the major and minor diameters of oocysts and the calculated morphometric index (P0.05. According to the elimination period, ducks eliminated oocysts for a longer period and in a higher number

  19. A new genotype of Cryptosporidium from giant panda (Ailuropoda melanoleuca) in China.

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  1. Distribution of Cryptosporidium and Giardia spp. in selected species of protected and game mammals from North-Eastern Poland.

    Science.gov (United States)

    Paziewska, Anna; Bednarska, Małgorzata; Niewegłowski, Hubert; Karbowiak, Grzegorz; Bajer, Anna

    2007-01-01

    Cryptosporidium spp. and Giardia spp. are wide-spread pathogens of humans and many species of mammals. The ways of transmission are very complex and difficult to define. Both parasites occur in similar environments and share a broad host range. However, in Poland there is still little known about the epidemiology of these parasites due to the paucity of data on human cases and only few studies in wildlife. The aim of our study was to determine the distribution of two intestinal protozoa in a few species of protected and game mammals in North-Eastern Poland. Additionally, we wanted to compare prevalence and abundance of these parasites between wild and farm animals, and to determine the species/genotypes of Cryptosporidium. Fecal samples collected from protected species (European beaver-22, grey wolf-14, European bison-55, Polish Konik (horse)-5) and game mammals (red deer-52, roe deer-22, boar-5) were examined by IFA. We also studied a group of samples collected from farm animals: beaver-30, red deer-66, Polish konik-5. Cryptosporidium oocysts were identified in 5 of 7 studied animal species (prevalence from 9% in roe deer to 36% in wolves), Giardia cysts in 4 of 6 studied species (prevalence from 1.7% in red deer to 7.7% in European beaver). Sequencing analysis of COWP gene fragment revealed that 5 Cryptosporidium isolates from wolves were C. parvum genotype 2 (zoonotic). The results show the important role of examined species in maintaining the natural sources of Cryptosporidium spp. and Giardia spp. infections in the environment.

  2. Classification of Cryptosporidium species from patients with sporadic cryptosporidiosis by use of sequence-based multilocus analysis following mutation scanning.

    Science.gov (United States)

    Jex, Aaron R; Pangasa, Aradhana; Campbell, Bronwyn E; Whipp, Margaret; Hogg, Geoff; Sinclair, Martha I; Stevens, Melita; Gasser, Robin B

    2008-07-01

    In the present study, we analyzed genetic variation in Cryptosporidium species from humans (n = 62) with clinical cryptosporidiosis in South Australia. Sequence variation was assessed in regions within the small subunit of nuclear rRNA (p-SSU), the 70-kDa heat shock protein (p-hsp70), and the 60-kDa glycoprotein (p-gp60) genes by employing single-strand conformation polymorphism analysis and sequencing. Based on the analyses of p-SSU and p-hsp70, Cryptosporidium hominis (n = 38) and Cryptosporidium parvum (n = 24) were identified. The analysis of p-gp60 revealed eight distinct subgenotypes, classified as C. hominis IaA17R1 (n = 3), IbA9G3R2 (n = 14), IbA10G2R2 (n = 20), and IfA12G1R1 (n = 1), as well as C. parvum IIaA18G3R1 (n = 15), IIaA20G3R1 (n = 6), IIaA22G4R1 (n = 2), and IIcA5G3R2 (n = 1). Subgenotypes IaA17R1 and IIaA22G4R1 are new. Of the six other subgenotypes, IbA10G2R2, IIaA18G3R1, IIaA20G3R1, and IIcA5G3R2 were reported previously from the state of Victoria. This is the fourth record in Australia of C. parvum subgenotype IIaA18G3R1 from humans, which, to date, has been isolated only from cattle in other countries. This subgenotype might be a significant contributor to sporadic human cryptosporidiosis and may indicate a greater zoonotic contribution to the infection of humans in the area of study. Comparative analyses revealed, for the first time, the differences in the genetic makeup of Cryptosporidium populations between two relatively close, major metropolitan cities.

  3. Glycoproteins and Gal-GalNAc cause Cryptosporidium to switch from an invasive sporozoite to a replicative trophozoite.

    Science.gov (United States)

    Edwinson, Adam; Widmer, Giovanni; McEvoy, John

    2016-01-01

    The apicomplexan parasite Cryptosporidium causes cryptosporidiosis, a diarrheal disease that can become chronic and life threatening in immunocompromised and malnourished people. There is no effective drug treatment for those most at risk of severe cryptosporidiosis. The disease pathology is due to a repeated cycle of host cell invasion and parasite replication that amplifies parasite numbers and destroys the intestinal epithelium. This study aimed to better understand the Cryptosporidium replication cycle by identifying molecules that trigger the switch from invasive sporozoite to replicative trophozoite. Our approach was to treat sporozoites of Cryptosporidium parvum and Cryptosporidium hominis, the species causing most human cryptosporidiosis, with various media under axenic conditions and examine the parasites for rounding and nuclear division as markers of trophozoite development and replication, respectively. FBS had a concentration-dependent effect on trophozoite development in both species. Trophozoite development in C. parvum, but not C. hominis, was enhanced when RPMI supplemented with 10% FBS (RPMI-FBS) was conditioned by HCT-8 cells for 3h. The effect of non-conditioned and HCT-8 conditioned RPMI-FBS on trophozoite development was abrogated by proteinase K and sodium metaperiodate pretreatment, indicating a glycoprotein trigger. Cryptosporidium parvum and C. hominis trophozoite development also was triggered by Gal-GalNAc in a concentration-dependent manner. Cryptosporidium parvum replication was greatest following treatments with Gal-GalNAc, followed by conditioned RPMI-FBS and non-conditioned RPMI-FBS (PGalNAc (1mM).

  4. An IC-PCR method for detection of Cryptosporidium and Giardia in natural surface waters in Finland.

    Science.gov (United States)

    Rimhanen-Finne, Ruska; Hörman, Ari; Ronkainen, Pilvi; Hänninen, Marja Liisa

    2002-08-01

    We developed an immunocapture-based polymerase chain reaction (PCR) assay for simultaneous detection of Cryptosporidium parvum oocysts and Giardia intestinalis cysts in surface water. Using primer pairs Cry9/Cry15 and LaxA/LaxB for Cryptosporidium and Gdh1/Gdh4 for Giardia, the sensitivity of the entire detection procedure (dealing with concentration, separation, DNA purification and PCR amplification) was at the level of 50-100 oocysts and cysts. Of 54 surface water samples, 4 were positive for Cryptosporidium and 1 for Giardia. Cryptosporidium and Giardia were detected for the first time in surface water in Finland.

  5. Some Aspects of Protozoan Infections in Immunocompromised Patients: A Review

    Directory of Open Access Journals (Sweden)

    Ferreira Marcelo Simão

    2002-01-01

    Full Text Available Protozoa are among the most important pathogens that can cause infections in immunocompromised hosts. These microorganisms particularly infect individuals with impaired cellular immunity, such as those with hematological neoplasias, renal or heart transplant patients, patients using high doses of corticosteroids, and patients with acquired immunodeficiency syndrome. The protozoa that most frequently cause disease in immunocompromised patients are Toxoplasma gondii, Trypanosoma cruzi, different Leishmania species, and Cryptosporidium parvum; the first two species cause severe acute meningoencephalitis and acute myocarditis, Leishmania sp. causes mucocutaneous or visceral disease, and Cryptosporidium can lead to chronic diarrhea with hepatobiliary involvement. Various serological, parasitological, histological and molecular methods for the diagnosis of these infections are currently available and early institution of specific therapy for each of these organisms is a basic measure to reduce the morbidity and mortality associated with these infections.

  6. Cryptosporidium and Giardia spp. infections in humans, animals and the environment in Poland.

    Science.gov (United States)

    Bajer, Anna

    2008-12-01

    Cryptosporidium spp. and Giardia spp. are intestinal protozoan parasites that are prevalent and widespread pathogens of humans and many other species of mammals. The aim of this review is to summarise the last 20 years of research on the epidemiology of these parasites, with a particular emphasis on the environment and the role played by different groups of animals in Poland. The prevalence of both species has been studied in different groups of humans, in wildlife, pets and farm animals and in environmental samples. Additionally, current knowledge on the distribution of zoonotic and non-zoonotic species/genotypes has been reviewed. The usefulness of different methods for the detection and identification of the parasites in different types of samples has been evaluated. Finally, because of the wide distribution and high prevalence of both species in a range of hosts and possible vectors involved in mechanical transmission, the overall risk of outbreaks of cryptosporidiosis and giardiosis in Poland has been assessed as relatively high.

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

    Science.gov (United States)

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

    2012-03-23

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

  8. Study of Cryptosporidium Infection in the Livestock (Cattle, Sheep, Dogs, Fowls and Humans, in Hamadan City and Its Suburbs during 2006-2011

    Directory of Open Access Journals (Sweden)

    H. Heidari

    2012-10-01

    Full Text Available Introduction & Objective: Cryptosporidium is one of the important pathogenic agents of gastero intestinal tract of mammals, especially human and domestic animals. This parasite is one of the principal causative agents of mortality in young animals. Considering the zoonotic importance of the parasite, infected animals can be very dangerous to the public health. This study was performed to determine the prevalence of cryptosporidial infection in humans, cattle, sheep, horses, dogs, domestic fowls and turkeys in Hamadan and its suburbs and also to evaluate the role of the animals of this area in transmission of the infection to human inhabitants of Hamadan. Materials & Methods: In this cross sectional descriptive study fecal samples were collected from 477 cattle, 220 sheep, 158 horses, 210 dogs, 200 domestic fowls, 200 domestic turkeys and 300 humans in Hamadan city. Cryptosporidium oocysts were concentrated by using the formalin-ether sedimentation method followed by the modified Ziehl-Neelsen staining technique and examined by light microcopy.Results: The prevalence rate of the infection in cattle, sheep, horses, dogs, domestic fowls, domestic turkeys and humans was 15.93%, 8.6%, 12.6%, 3.8%, 2.5%, 4% and 5.33%, respectively. Conclusion: Cryptosporidium can cause diarrhea by itself or with other microbial agents in inhabitants of Hamadan. Considering the high prevalence and intensity of the infection in the cattle of the suburbs of Hamadan and with regard to the results of the other researches in the world and Iran, cattle are one of the most important sources of the infection for the inhabitants of Hamadan.(Sci J Hamadan Univ Med Sci 2012;19(3:67-74

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

    Science.gov (United States)

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

    2016-01-01

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

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

    Science.gov (United States)

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

    2016-08-01

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

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

    Science.gov (United States)

    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 (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.

  12. The effects of Moringa lieifera seed powder on turbidity and sedimentation of Cryptosporidium spp. in wastewater

    DEFF Research Database (Denmark)

    Petersen, H. H.; Wolsey, I.; Dalsgaard, A.

    2013-01-01

    or water used for postharvest washing of the produce is contaminated. A laboratory study was carried out to investigate the effect of a coagulant from the seeds of Moringa oleifera (MO) in reducing Cryptosporidium parvum oocysts and turbidity in Danish wastewater. To each of five time points, 12 replicates...

  13. INFECTIVITY OF A CRYPTOSPORIDIUM PARVUM ISOLATE OF CERVINE ORIGIN FOR HEALTHY ADULTS AND GAMMA INTERFERON-G KNOCKOUT MICE. (R828035)

    Science.gov (United States)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  14. INFECTIVITY OF A CRYPTOSPORIDIUM PARVUM ISOLATE OF CERVINE ORIGIN FOR HEALTHY ADULTS AND GAMMA INTERFERON-G KNOCKOUT MICE. (R829180)

    Science.gov (United States)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  15. INFECTIVITY OF A CRYPTOSPORIDIUM PARVUM ISOLATE OF CERVINE ORIGIN FOR HEALTHY ADULTS AND GAMMA INTERFERON-G KNOCKOUT MICE. (R828035)

    Science.gov (United States)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  16. INFECTIVITY OF A CRYPTOSPORIDIUM PARVUM ISOLATE OF CERVINE ORIGIN FOR HEALTHY ADULTS AND GAMMA INTERFERON-G KNOCKOUT MICE. (R829180)

    Science.gov (United States)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  17. Occurrence and molecular characterization of Giardia duodenalis and Cryptosporidium spp. in sheep and goats reared under dairy husbandry systems in Greece☆

    Science.gov (United States)

    Tzanidakis, Nikolaos; Sotiraki, Smaragda; Claerebout, Edwin; Ehsan, Amimul; Voutzourakis, Nikolaos; Kostopoulou, Despoina; Stijn, Casaert; Vercruysse, Jozef; Geurden, Thomas

    2014-01-01

    Giardia duodenalis and Cryptosporidium spp. are gastro-intestinal protozoa known to infect small ruminants. Both protozoa are also considered as a potential public health concern. The objective of this study was to determine their prevalence in lambs and goat kids kept under common Mediterranean dairy husbandry systems and to identify the species and genotypes infecting these small ruminants. In total, 684 faecal samples (429 from lambs and 255 from goat kids) were collected on 21 farms in Greece and examined using a quantitative immunofluorescence assay. G. duodenalis was detected in 37.3% of the lambs and 40.4% of the goat kids. On all but one of the farms G. duodenalis was detected. Most samples were typed as a mono-infection with G. duodenalis assemblage E, both on the β-giardin gene and the triose phosphate isomerase gene. Only 10% of samples were typed as mixed assemblage A and E infections. The prevalence of Cryptosporidium spp. was 5.1% in lambs and 7.1% in goat kids. In total, 8 out of the 14 farms with a sheep flock and 7 out of the 14 farms with a goat flock were positive. Cryptosporidium parvum (subtype IId), C. ubiquitum and C. xiaoi were identified, the latter especially in goat kids. In conclusion, the results of the present study illustrate that G. duodenalis and Cryptosporidium spp. occur frequently on both sheep and goats farms. The prevalence of zoonotic genotypes or species was low, indicating a limited but existing risk for zoonotic infections. PMID:25187088

  18. Host and environmental risk factors associated with Cryptosporidium scophthalmi (Apicomplexa) infection in cultured turbot, Psetta maxima (L.) (Pisces, Teleostei).

    Science.gov (United States)

    Alvarez-Pellitero, Pilar; Perez, Andrés; Quiroga, M Isabel; Redondo, M José; Vázquez, Sonia; Riaza, Ana; Palenzuela, Oswaldo; Sitjà-Bobadilla, Ariadna; Nieto, José M

    2009-11-12

    An epidemiological cohort study of Cryptosporidium scophthalmi in cultured turbot Psetta maxima L. of Northwestern Spain was conducted along a four-year period. Four different ongrowing cohorts were monitored monthly from introduction into the ongrowing tanks (10-50 g) until reaching market size (400-1400 g). The association of host and environmental factors with five categories of parasite abundance was assessed using a multivariable regression framework. Epidemiological factors assessed here were water temperature, weight, length, month of collection, season, age, origin, condition factor, water filtration, and status to the myxozoan Enteromyxum scophthalmi infection. E. scophthalmi was included into the analysis because it targets the same organ than C. scophthalmi and it was prevalent in the studied population. The multivariable analysis demonstrated the statistically significant association between several factors and parasite abundance. C. scophthalmi abundance was associated (P<0.05) with age, condition factor, season, and status to E. scophthalmi infection. Young animals, with poor condition factor, during spring or summer, and not infected with the myxozoan were most likely to be highly infected by C. scophthalmi. Inclusion of these four variables significantly (P<0.05) improved the model, compared to the model that did not include any of these epidemiological factors. Increasing levels of C. scophthalmi abundance were associated (P<0.01) with higher severity of C. scophthalmi-compatible lesions. The frequency of distribution of C. scophthalmi abundance was clearly right-skewed and fitted a negative binomial distribution, whereas the intensity of infection fitted a Poisson distribution. The quantification of the variance-to-mean ratio stratified by age demonstrated overdispersion for 8-16 months old fish, although this bivariate association is likely affected by several other factors, as suggested by the results of the multivariable analysis. The negative

  19. The General Biology of Cryptosporidium

    Science.gov (United States)

    The protozoan parasites belonging to the genus Cryptosporidium infect all classes of vertebrate animals. Of the sixteen valid species and nearly forty genotypes, three species are responsible for the majority of economic losses to livestock and morbidity and mortality to humans. This chapter summari...

  20. Validation of multiple diagnostic techniques to detect Cryptosporidium sp. and Giardia sp. in free-ranging western lowland gorillas (Gorilla gorilla gorilla) and observations on the prevalence of these protozoan infections in two populations in Gabon.

    Science.gov (United States)

    van Zijll Langhout, Martine; Reed, Patricia; Fox, Mark

    2010-06-01

    Anthropozoonotic diseases threaten the survival of western lowland gorillas (Gorilla gorilla gorilla). Use of accurate diagnostic techniques in gorilla health monitoring contributes to the conservation of gorillas by providing robust information for appropriate management decisions. To identify suitable protozoa diagnostic techniques for wild gorillas, 95 fecal specimens were collected in Lopé National Park and east of Moukalaba-Doudou National Park in Gabon, areas with high and low levels of human activity, respectively. The samples were examined for Cryptosporidium sp. and Giardia sp. by using the following diagnostic techniques: a commercially available immunofluorescent antibody test kit, Merifluor, and a rapid immune-assay, ImmunoCard STAT!, to detect Cryptosporidium sp. and Giardia sp., and a modified Ziehl-Neelsen stain to detect Cryptosporidium sp. oocysts. The results obtained from the Merifluor test, considered the "gold standard" in human studies, were used to estimate the prevalence of Cryptosporidium sp. and Giardia sp. infections in Lopé National Park (19.0% and 22.6%, respectively) and east of Moukalaba-Doudou National Park (0% and 9.1%, respectively). The difference in prevalence in both areas may be associated with differing levels of anthropogenic disturbance. The sensitivity and specificity of the latter two diagnostic techniques were calculated by using the Merifluor test as a control. The ImmunoCard STAT! was found suitable for Giardia sp. antigen detection (specific but not sensitive) and inappropriate for Cryptosporidium sp. antigen detection (not specific or sensitive). The modified Ziehl-Neelsen stain was found to be highly specific but not sensitive in the detection of Cryptosporidium sp. oocysts. These results underline the necessity of using ancillary tests and concentration methods to correctly identify positive samples. This is the first report of Cryptosporidium sp. and Giardia sp. infections in free-ranging western lowland gorillas

  1. Molecular characterization of Cryptosporidium spp. from fecal samples of birds kept in captivity in Brazil.

    Science.gov (United States)

    Nakamura, Alex Akira; Simões, Daniel Castendo; Antunes, Rômulo Godik; da Silva, Deuvânia Carvalho; Meireles, Marcelo Vasconcelos

    2009-12-03

    The aim of this study was to determine the prevalence of Cryptosporidium species and genotypes in birds kept in captivity in Brazil. A total of 966 samples from 18 families of birds was collected and stored in 5% potassium dichromate solution at 4 degrees C until processing. Oocysts were purified in Sheather sugar solution following extraction of genomic DNA. Molecular analyses were performed using nested-PCR for amplification of fragments of the 18S subunit of rRNA gene and of the actin gene. Amplification of Cryptosporidium DNA fragments was obtained in 47 (4.86%) samples. Sequencing of amplified fragments and phylogenetic analyses allowed the identification of Cryptosporidium baileyi in a black vulture (Coragyps atratus), a domestic chicken (Gallus gallus domesticus) and a saffron finch (Sicalis flaveola); Cryptosporidium galli in canaries (Serinus canaria), a cockatiel (Nymphicus hollandicus) and lesser seed-finches (Oryzoborus angolensis); Cryptosporidium meleagridis in a domestic chicken (G. g. domesticus); Cryptosporidium parvum in a cockatiel (N. hollandicus); Cryptosporidium avian genotype I in a canary (S. canaria) and an Indian peafowl (Pavo cristatus); Cryptosporidium avian genotype II in ostriches (Struthio camelus) and Cryptosporidium avian genotype III in a cockatiel (N. hollandicus) and a peach-faced lovebird (Agapornis roseicolis).

  2. Detection of concurrent infection of dairy cattle with Blastocystis, Cryptosporidium, Giardia, and Enterocytozoon by molecular and microscopic methods

    Science.gov (United States)

    Of fecal specimens examined from 47 dairy cattle ranging in age from neonates to multiparous cows, 9, 10, 24, and 17 were positive for Blastocystis spp., Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi, respectively, as determined by PCR. Eight 3- to 5-month-old cattle were co...

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

    Science.gov (United States)

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

    2013-11-08

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

  4. Cryptosporidium spp. infection in mares and foals of the northwest region of São Paulo State, Brazil Infecção por Cryptosporidium spp. em éguas e potros da região noroeste do estado de São Paulo, Brasil

    Directory of Open Access Journals (Sweden)

    Sandra Valéria Inácio

    2012-12-01

    Full Text Available The present study aimed to analyze the occurrence of infection by Cryptosporidium spp. in mares and their respective foals. This study was carried out in 11 farms located in the municipalities of Araçatuba, Birigui, Guararapes and Santo Antônio do Aracangua, in the northwest region of the State of Sao Paulo, from November 2010 to March 2011. A total of 98 mares and 98 foals of several breeds were analyzed; among foals, 59 were males and 39 females, aged from three to 330 days. Feces were collected directly from the rectal ampulla, purified and processed according to modified Kinyoun stain. Occurrence of Cryptosporidium spp. was 21.4% (21/98 for foals and 18.4% (18/98 for mares. Occurrence of Cryptosporidium spp. had significant association with breeds and age of animals. Results obtained led to the conclusion that foals older than two months and Mangalarga animals are less susceptible to the occurrence of Cryptosporidium spp.O presente estudo teve como objetivo analisar a ocorrência da infecção por Cryptosporidium spp. em éguas e seus respectivos potros. Este estudo foi realizado em 11 fazendas localizadas nos municípios de Araçatuba, Birigui, Guararapes e Santo Antônio do Aracangua, na região Noroeste do Estado de São Paulo, de novembro de 2010 a março de 2011. Um total de 98 éguas e 98 potros de diversas raças foram analisados, sendo que, entre os filhotes, 59 eram machos e 39 fêmeas, cujas idades variavam de três até 330 dias. Fezes foram colhidas diretamente da ampola retal, purificadas e processadas pela técnica de Kinyoun modificada. A ocorrência de Cryptosporidium spp. observada foi de 21,4% (21/98 para potros e 18,4% (18/98 para éguas. A ocorrência de Cryptosporidium spp. teve uma associação significativa com a raça e a idade dos animais. A partir dos resultados obtidos, conclui-se neste estudo que potros com idade superior a dois meses e animais da raça Mangalarga foram menos susceptíveis à ocorrência de

  5. Molecular Insights for Giardia, Cryptosporidium, and Soil-Transmitted Helminths from a Facility-Based Surveillance System in Guatemala

    Science.gov (United States)

    Velasquez, Daniel E.; Arvelo, Wences; Cama, Vitaliano A.; López, Beatriz; Reyes, Lissette; Roellig, Dawn M.; Kahn, Geoffrey D.; Lindblade, Kimberly A.

    2011-01-01

    We molecularly characterized samples with Giardia, Cryptosporidium, and soil-transmitted helminths from a facility-based surveillance system for diarrhea in Santa Rosa, Guatemala. The DNA sequence analysis determined the presence of Giardia assemblages A (N = 7) and B (N = 12) and, Cryptosporidium hominis (N = 2) and Cryptosporidium parvum (N = 2), suggestive of different transmission cycles. All 41 samples with soil-transmitted helminths did not have the β-tubulin mutation described for benzimidazole resistance, suggesting potential usefulness in mass drug administration campaigns. PMID:22144459

  6. Bioaccumulation of Toxoplasma and Cryptosporidium by the crustacean Gammarus fossarum: involvement in biomonitoring survey and trophic transfer

    Science.gov (United States)

    The protozoans Toxoplasma gondii and Cryptosporidium parvum are public health priorities and their oocysts can persist in environment for long time. They are present in various watercourses as recreational, surface, drinking, river, and seawater and could interact with organisms. To evaluate the cap...

  7. A multi-locus study of cryptosporidium parasites isolated from patients living in iran, Malawi, Nigeria, the United kingdom, and Vietnam.

    Directory of Open Access Journals (Sweden)

    Salman Ghaffari

    2014-03-01

    Full Text Available Cryptosporidium species are important cause of diarrheal diseases in both developing and developed countries. This study aimed to compare the performance of several molecular methods for identification of Cryptosporidium species, and to detect genetic variation among each of these species isolated from Iran, Malawi, Nigeria, Vietnam and the United Kingdom.The oocysts DNA samples were derived from 106 Cryptosporidium positive feces. Polymerase chain reaction, PCR- restriction fragment length polymorphism and DNA sequence analysis of the 18S rRNA and the Cryptosporidium oocysts wall protein genes; PCR and DNA sequence analysis of a fragment of 70 kDa heat shock protein and 60 kDa glycoprotein genes were carried out.Based on these analysis, three species of Cryptosporidium including C. hominis, C. parvum and C. meleagridis, and both C. hominis and C. parvum were found in Iranian and the UK samples, respectively. Also, three C. hominis (Ib, Ib3& Id and three C. parvum (IIa, IIc & IId subtypes were identified by sequence analysis of the GP60 gene. Of these, C. hominis Ib was predominant and interestingly, one subgenotype (C. hominis Ib A10G2 accounted for the majority of the samples.The current study demonstrates the complex subtypes of Cryptosporidium isolates in both developing and developed countries. This is the first report of C. parvum IId subgenotype and three new subtypes of C. parvum IIa in the UK, a new subtype of C. hominis Id from Malawi; and the first multi-locus study of three species of Cryptosporidium in human from Iran.

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

    Science.gov (United States)

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

    2016-10-14

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

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

    Science.gov (United States)

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

    2015-06-01

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

  10. Prevalence and genetic characteristics of Cryptosporidium, Enterocytozoon bieneusi and Giardia duodenalis in cats and dogs in Heilongjiang province, China.

    Science.gov (United States)

    Li, Wei; Li, Yijing; Song, Mingxin; Lu, Yixin; Yang, Jinping; Tao, Wei; Jiang, Yanxue; Wan, Qiang; Zhang, Siwen; Xiao, Lihua

    2015-03-15

    This study investigated 319 fecal specimens of cats (n=52) and dogs (n=267) from Heilongjiang province, China for the prevalence and genetic characteristics of Cryptosporidium, Enterocytozoon bieneusi, and Giardia duodenalis. PCR and DNA sequence analysis of the small subunit rRNA gene identified C. felis and C. parvum in one cat each (3.8%) and C. canis and C. ubiquitum in 6 dogs (2.2%). Polymorphisms in the ribosomal internal transcribed spacer and phylogenetic analysis characterized zoonotic E. bieneusi genotypes D, EbpC, NED1, and NED2 and host-adapted ones NED3, NED4, and PtEb IX in 18 dogs (6.7%) and human-pathogenic genotypes D and IV in 3 cats (5.8%). Genotyping based on the hypermutation of G. duodenalis triosephosphate isomerase gene (TPI) facilitated identification of assemblage F in a cat (1.9%) and assemblages C and E in 12 dogs (4.5%). Subtypes of G. duodenalis isolates were determined by measuring the diversity of both TPI nucleotide and amino acid sequences. C. canis, C. felis, C. parvum, E. bieneusi genotypes D, EbpC, and IV, and G. duodenalis assemblage C identified herein have been documented in human infections in China. C. canis, C. parvum, C. ubiquitum, and E. bieneusi genotypes D, EbpC, and IV carried by cats or dogs also existed in wastewater in China. The finding suggested pet animals could be reservoirs for human cryptosporidiosis, microsporidiosis, and giardiasis and potential sources of water contamination in China.

  11. Molecular appraisal of intestinal parasitic infection in transplant recipients

    Science.gov (United States)

    Yadav, Pooja; Khalil, Shehla; Mirdha, Bijay Ranjan

    2016-01-01

    Background & objectives: Diarrhoea is the main clinical manifestation caused by intestinal parasitic infections in patients, with special reference to transplant recipients who require careful consideration to reduce morbidity and mortality. Further, molecular characterization of some important parasites is necessary to delineate the different modes of transmission to consider appropriate management strategies. We undertook this study to investigate the intestinal parasitic infections in transplant recipients with or without diarrhoea, and the genotypes of the isolated parasites were also determined. Methods: Stool samples from 38 transplant recipients comprising 29 post-renal, two liver and seven bone marrow transplant (BMT) recipients presenting with diarrhoea and 50 transplant recipients (42 post-renal transplant, eight BMT) without diarrhoea were examined for the presence of intestinal parasites by light microscopy using wet mount, modified Ziehl–Neelsen staining for intestinal coccidia and modified trichrome staining for microsporidia. Genotypes of Cryptosporidium species were determined by multilocus genotyping using small subunit ribosomal (SSUrRNA), Cryptosporidium oocyst wall protein (COWP) and dihydrofolate reductase (DHFR) as the target genes. Assemblage study for Giardia lamblia was performed using triose phosphate isomerase (TPI) as the target gene. Samples were also screened for bacterial, fungal and viral pathogens. Results: The parasites that were detected included Cryptosporidium species (21%, 8/38), Cystoisospora (Isospora) belli (8%, 3), Cyclospora cayetanensis (5%, 2), G. lamblia (11%, 4), Hymenolepis nana (11%, 4), Strongyloides stercoralis (3%, 1) and Blastocystis hominis (3%, 1). Multilocus genotyping of Cryptosporidium species at SSUrRNA, COWP and DHFR loci could detect four isolates of C. hominis; two of C. parvum, one of mixed genotype and one could not be genotyped. All the C. hominis isolates were detected in adult post

  12. Molecular appraisal of intestinal parasitic infection in transplant recipients

    Directory of Open Access Journals (Sweden)

    Pooja Yadav

    2016-01-01

    Full Text Available Background & objectives: Diarrhoea is the main clinical manifestation caused by intestinal parasitic infections in patients, with special reference to transplant recipients who require careful consideration to reduce morbidity and mortality. Further, molecular characterization of some important parasites is necessary to delineate the different modes of transmission to consider appropriate management strategies. We undertook this study to investigate the intestinal parasitic infections in transplant recipients with or without diarrhoea, and the genotypes of the isolated parasites were also determined. Methods: Stool samples from 38 transplant recipients comprising 29 post-renal, two liver and seven bone marrow transplant (BMT recipients presenting with diarrhoea and 50 transplant recipients (42 post-renal transplant, eight BMT without diarrhoea were examined for the presence of intestinal parasites by light microscopy using wet mount, modified Ziehl-Neelsen staining for intestinal coccidia and modified trichrome staining for microsporidia. Genotypes of Cryptosporidium species were determined by multilocus genotyping using small subunit ribosomal (SSUrRNA, Cryptosporidium oocyst wall protein (COWP and dihydrofolate reductase (DHFR as the target genes. Assemblage study for Giardia lamblia was performed using triose phosphate isomerase (TPI as the target gene. Samples were also screened for bacterial, fungal and viral pathogens. Results: The parasites that were detected included Cryptosporidium species (21%, 8/38, Cystoisospora (Isospora belli (8%, 3, Cyclospora cayetanensis (5%, 2, G. lamblia (11%, 4, Hymenolepis nana (11%, 4, Strongyloides stercoralis (3%, 1 and Blastocystis hominis (3%, 1. Multilocus genotyping of Cryptosporidium species at SSUrRNA, COWP and DHFR loci could detect four isolates of C. hominis; two of C. parvum, one of mixed genotype and one could not be genotyped. All the C. hominis isolates were detected in adult post

  13. Molecular characterization of Cryptosporidium and Giardia in farmers and their ruminant livestock from the Coastal Savannah zone of Ghana.

    Science.gov (United States)

    Squire, Sylvia Afriyie; Yang, Rongchang; Robertson, Ian; Ayi, Irene; Ryan, Una

    2017-09-21

    Cryptosporidium and Giardia are major causes of diarrhoea in developing countries including Ghana, however, nothing is known about the species and subtypes of Cryptosporidium and Giardia in farmers and their ruminant livestock in this country. A total of 925 faecal samples from humans (n=95), cattle (n=328), sheep (n=217) and goats (n=285), were screened for Cryptosporidium and Giardia by quantitative PCR (qPCR) at the 18S rRNA and glutamate dehydrogenase (gdh) loci respectively. Cryptosporidium positives were typed by sequence analysis of 18S and 60kDa glycoprotein (gp60) loci amplicons. Giardia positives were typed at the triose phosphate isomerase (tpi), beta-giardin (bg) and gdh loci. The prevalence of Cryptosporidium and Giardia by qPCR was 8.4% and 10.5% in humans, 26.5% and 8.5% in cattle, 34.1% and 12.9% in sheep, and 33.3% and 12.3% in goat faecal samples, respectively. G. duodenalis assemblages A and B were detected in humans and assemblage E was detected in livestock. Cryptosporidium parvum was the only species identified in humans; C. andersoni, C. bovis, C. ryanae and C. ubiquitum were identified in cattle; C. xiaoi, C. ubiquitum and C. bovis in sheep; and C. xiaoi, C. baileyi and C. parvum in goats. This is the first molecular study of Cryptosporidium and Giardia in livestock in Ghana. The identification of zoonotic species and the identification of C. parvum subtype IIcA5G3q in livestock, which has previously been identified in children in Ghana, suggests potential zoonotic transmission. Further studies on larger numbers of human and animal samples, and on younger livestock are required to better understand the epidemiology and transmission of Cryptosporidium and Giardia in Ghana. Copyright © 2017. Published by Elsevier B.V.

  14. A new set of primers directed to 18S rRNA gene for molecular identification of Cryptosporidium spp. and their performance in the detection and differentiation of oocysts shed by synanthropic rodents.

    Science.gov (United States)

    Silva, Sheila O S; Richtzenhain, Leonardo J; Barros, Iracema N; Gomes, Alessandra M M C; Silva, Aristeu V; Kozerski, Noemila D; de Araújo Ceranto, Jaqueline B; Keid, Lara B; Soares, Rodrigo M

    2013-11-01

    Cryptosporidium spp. are cosmopolitan protozoa that infect fishes, reptiles, amphibians, birds and mammals. More than 20 species are recognized within this genus. Rodents are a group of abundant and ubiquitous organisms that have been considered reservoirs of Cryptosporidium for humans and livestock. The aim of this study was to design specific primers for the gene encoding 18S rRNA, potentially capable of amplifying any species or genotype of Cryptosporidium spp. and evaluate the diagnostic attributes of the nested-PCR based on such probes. The primers were designed to amplify the shortest segment as possible to maximize the sensitivity of the test, but preserving the discriminatory potential of the amplified sequences for phylogenetic inferences. The nested-PCR standardized in this study (nPCR-SH) was compared in terms of sensitivity with another similar assay (nPCR-XIAO) that has been largely used for the detection and identification of Cryptosporidium spp. worldwide. We also aimed to molecularly characterize samples of Cryptosporidum spp. isolated from synanthropic rodents using these probes. Forty-five rodents were captured in urban areas of the municipality of Umuarama, Paraná State, Brazil. Fecal samples were submitted to three molecular tests (nested-PCRs), two of them targeted to the 18S rDNA gene (nPCR-SH and nPCR-XIAO) and the third targeted to the gene encoding actin (nPCR-actin). The nPCR-SH was tested positive on samples of Cryptosporidum parvum, Cryptosporidum andersoni, Cryptosporidum meleagridis, Cryptosporidum hominis, Cryptosporidum canis, and Cryptosporidum serpentis. Sixteen samples of rodents were positive by nPCR-SH, six by nPCR-XIAO and five by nPCR-actin. Sequencing of amplified fragments allowed the identification of Cryptosporidum muris in three samples of Rattus rattus, and two genotypes of Cryptosporidium, the genotypes mouse II and III. Cryptosporidium genotype mouse II was found in one sample of Mus musculus and genotype mouse III

  15. Infections by protozoa in immunocompromised hosts

    Directory of Open Access Journals (Sweden)

    Ferreira Marcelo Simão

    2000-01-01

    Full Text Available Protozoa are among the most important pathogens that can cause infection in immunocompromised patients. They infect particularly individuals with impaired cell immunity, such as those with hematologic neoplasias, those submitted to transplant of solid organs, those under high-dose corticosteroid therapy, and carriers of the human immunodeficiency virus. Among the protozoa that most commonly cause disease in immunocompromised individuals are Toxoplasma gondii, Trypanosoma cruzi, Cryptosporidium parvum, Isospora belli, Cyclospora cayetanensis and microsporidia; the former two cause severe encephalitis and myocarditis, and the others cause gastrointestinal infections. Early diagnosis and prompt institution of specific therapy for each of these organisms are basic measures to decrease morbidity and mortality associated with these infections.

  16. Concurrent infections with Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi, and Blastocystis spp. in naturally infected dairy cattle from birth to two years of age

    Science.gov (United States)

    Fecal specimens were collected directly at weekly and then monthly intervals from each of 30 dairy calves from birth to 24 months to determine the prevalence and age distribution of Cryptosporidium spp., Giardia duodenalis assemblages, Enterocytozoon bieneusi genotypes, and Blastocystis spp subtypes...

  17. A Longitudinal Study of Cryptosporidium, Giardia, and Enterocytozoon Bieneusi Infections in Naturally Infected Dairy Cattle from Birth to Two Years of Age

    Science.gov (United States)

    Fecal specimens were collected from 30 calves from birth to 24 months of age at a dairy farm in Maryland to determine the prevalence and age distribution of Cryptosporidium, Giardia, Enterocytozoon bieneusi species/genotypes. It is crucial to know which species and/or genotypes are actually present ...

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

    Science.gov (United States)

    Nasser, Abidelfatah M

    2016-02-01

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

  19. Identification and differentiation of Cryptosporidium species by capillary electrophoresis single-strand conformation polymorphism.

    Science.gov (United States)

    Power, Michelle L; Holley, Marita; Ryan, Una M; Worden, Paul; Gillings, Michael R

    2011-01-01

    Cryptosporidium species generally lack distinguishing morphological traits, and consequently, molecular methods are commonly used for parasite identification. Various methods for Cryptosporidium identification have been proposed, each with their advantages and disadvantages. In this study, we show that capillary electrophoresis coupled with single-strand conformation polymorphism (CE-SSCP) is a rapid, simple and cost-effective method for the identification of Cryptosporidium species and genotypes. Species could be readily differentiated based on the SSCP mobility of amplified 18S rRNA gene molecules. Clones that differed by single-nucleotide polymorphisms could be distinguished on CE-SSCP mobility. Profiles of species known to have heterogenic copies of 18S rRNA gene contained multiple peaks. Cloning and sequencing of Cryptosporidium parvum, Cryptosporidium hominis, Cryptosporidium fayeri and Cryptosporidium possum genotype 18S rRNA gene amplicons confirmed that these multiple peaks represented type A and type B 18S rRNA gene copies. CE-SSCP provides a reliable and sensitive analysis for epidemiological studies, environmental detection and diversity screening.

  20. Probabilistic quantitative microbial risk assessment model of farmer exposure to Cryptosporidium spp. in irrigation water within Kumasi Metropolis-Ghana

    DEFF Research Database (Denmark)

    Sampson, Angelina; Owusu-Ansah, Emmanuel de-Graft Johnson; Mills-Robertson, Felix C.

    2017-01-01

    Cryptosporidium is a protozoan parasite which can be transmitted via food and water. Some studies have shown irrigation water to be routes of transmission for Cryptosporidium into the food chain, however, little information is known about Cryptosporidium levels in wastewater used for irrigation...... causing gastroenteritis. The results indicate high positive levels of Cryptosporidium in the irrigation water, however, the levels of Cryptosporidium decreases during the rainfall seasons, risk assessment results show that, farmers face a higher risk of being infected by Cryptosporidium due to frequent...

  1. Adhesive-tape recovery combined with molecular and microscopic testing for the detection of Cryptosporidium oocysts on experimentally contaminated fresh produce

    Science.gov (United States)

    Cryptosporidium parvum suspended in water were applied to the surface of apples, cucumbers, peaches, and tomatoes at concentrations of 100, 50 or 10 oocysts within circles drawn with a permanent marker. Approximately 18 hr later, skin containing uncontaminated and contaminated circles from each prod...

  2. Molecular Epidemiology of Giardia, Blastocystis and Cryptosporidium among Indigenous Children from the Colombian Amazon Basin

    Science.gov (United States)

    Sánchez, Angie; Munoz, Marina; Gómez, Natalia; Tabares, Juan; Segura, Laura; Salazar, Ángela; Restrepo, Cristian; Ruíz, Miguel; Reyes, Patricia; Qian, Yuchen; Xiao, Lihua; López, Myriam C.; Ramírez, Juan D.

    2017-01-01

    The incidence and prevalence of intestinal parasites in children is most likely due to lack of natural or acquired resistance and differences in behavior and habits closely related to environmental and socioeconomic determinants. The most important protozoa that parasitize humans are Giardia, Entamoeba, Blastocystis, and Cryptosporidium. These parasites present wide intraspecific genetic diversity and subsequently classified into assemblages and subtypes. The Amazon basin is the largest in the world and is the fifth freshwater reserve on the planet. Contradictorily, people living in these areas (Indigenous populations) have poor quality of life, which favors the infection of diseases of fecal-oral transmission. The aim of this work was to unravel the molecular epidemiology of Giardia, Blastocystis and Cryptosporidium across four communities (Puerto Nariño, San Juan del Soco, Villa Andrea and Nuevo Paraíso). We obtained 284 fecal samples from children under 15 years old that were analyzed by direct microscopy (261 samples) and Real Time PCR (qPCR) (284 samples). The positive samples for these protozoa were further characterized by several molecular markers to depict assemblages and subtypes. We observed a frequency of Giardia infection by microscopy of 23.7% (62 samples) and by qPCR of 64.8% (184 samples); for Blastocystis by microscopy of 35.2% (92 samples) and by qPCR of 88.7% (252 samples) and for Cryptosporidium only 1.9% (5 samples) were positive by microscopy and qPCR 1.8% (5 samples). Regarding the Giardia assemblages, using the glutamate dehydrogenase (gdh) marker we observed AI, BIII and BIV assemblages and when using triose phosphate isomerase (tpi) we observed assemblages AI, AII, BIII and BIV. In contrast, Blastocystis STs detected were 1, 2, 3, 4, and 6. Lastly, the species C. viatorum, C. hominis (with the subtypes IdA19 and IaA12R8) and C. parvum (with the subtype IIcA5G3c) were identified. We observed a high profile of zoonotic transmission

  3. 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 Ocorrência e caracterização molecular de Cryptosporidium spp. isolados de animais domésticos de propriedades rurais circunvizinhas a fragmentos de Floresta Atlântica Seca do Estado de São Paulo, Brasil

    Directory of Open Access Journals (Sweden)

    Anaiá da Paixão Sevá

    2010-12-01

    Full Text Available 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.O objetivo deste estudo foi avaliar a ocorrência de Cryptosporidium, em animais domésticos de propriedades rurais ao redor de fragmentos de mata Atlântica de interior no município de Teodoro Sampaio, por exame convencional de flutuação em solução de sacarose, seguido de caracterização molecular

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

    Science.gov (United States)

    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

  5. Occurrence of Giardia and Cryptosporidium in wild birds in Galicia (Northwest Spain).

    Science.gov (United States)

    Reboredo-Fernández, Aurora; Ares-Mazás, Elvira; Cacciò, Simone M; Gómez-Couso, Hipólito

    2015-06-01

    Faecal samples were obtained from 433 wild birds being treated in wildlife recovery centres in Galicia (Northwest Spain), between February 2007 and September 2009. The birds belonged to 64 species representing 17 different orders. Giardia cysts and Cryptosporidium oocysts were detected by an immunofluorescence antibody test and identified at the molecular level by established PCR-sequencing methods. The overall prevalence of Giardia was 2·1% and that of Cryptosporidium, 8·3%. To our knowledge, this is the first description of Giardia sp. in Tyto alba and Caprimulgus europaeus; and of Cryptosporidium sp. in Apus apus, Athene noctua, C. europaeus, Falco tinnunculus, Morus bassanus, Parabuteo unicinctus and Strix aluco. Furthermore, the first PCR-sequence confirmed detection of Giardia duodenalis assemblage B in, Buteo buteo, Coturnix coturnix and Pica pica; G. duodenalis assemblage D in Garrulus glandarius; and G. duodenalis assemblage F in Anas platyrhynchos; Cryptosporidium parvum in Accipiter nisus, B. buteo, Milvus migrans, Pernis apivorus and P. pica; and Cryptosporidium meleagridis in Streptopelia turtur. The study findings demonstrate the wide spread of Giardia and Cryptosporidium between wild birds.

  6. Molecular genotyping and sub-genotyping of Cryptosporidium spp. isolates from symptomatic individuals attending two major public hospitals in Madrid, Spain.

    Science.gov (United States)

    de Lucio, Aida; Merino, Francisco J; Martínez-Ruiz, Rocío; Bailo, Begoña; Aguilera, María; Fuentes, Isabel; Carmena, David

    2016-01-01

    Infections by members of the protozoan genus Cryptosporidium are among the most common causes of human gastrointestinal illness worldwide. In Spain cryptosporidiosis is not a compulsory notifiable disease, so the actual burden of the infection in both clinical and general populations remains largely unknown. We present here data on the diversity and frequency of the Cryptosporidium species and sub-genotypes identified in symptomatic individuals seeking medical care in two major hospitals in Madrid, Spain, between December 2013 and January 2015. Initial detection of the parasite was conducted on a total of 122 stool samples collected from 120 patients by microscopy with modified Ziehl-Neelsen and/or immunochromatographic tests. We used immunofluorescence, PCR-based methods and sequence analyses of the 60-kDa (GP60) glycoprotein and the small subunit ribosomal RNA (SSU rRNA) genes for confirmatory purposes and to characterize Cryptosporidium isolates. A total of 110 patients were confirmed with cryptosporidiosis. Overall, 101 isolates were successfully sub-genotyped at the GP60 locus, and an additional seven at the SSU rRNA locus. The analyses of all amplicons defined 10 distinct sequence types representing the GP60 family sub-genotypes IbA10G2 (78.7%), IeA11G3T3 (3.7%) of C. hominis, and the GP60 family sub-types IIaA15G2R1 (5.6%), IIaA18G6R1 (0.9%), IIcA5G3a (0.9%), IIdA18G1 (0.9%), IIdA19G1 (0.9%), IIdA21G1 (0.9%), and IIdA22G1 (0.9%) of C. parvum. A single isolate was assigned to C. felis (0.9%), two C. parvum isolates (1.9%) could not be characterized at the sub-genotype level and an additional four isolates (3.7%) were not typable. These results strongly suggest that transmission of cryptosporidiosis is mostly anthroponotic in origin in the clinical sample under study. We expect that our molecular epidemiological data will make a significant contribution to unravel the actual epidemiological situation of cryptosporidiosis in Spain, providing health care and

  7. Ruling out nosocomial transmission of Cryptosporidium in a renal transplantation unit: case report.

    Science.gov (United States)

    Brunet, J; Lemoine, J P; Pesson, B; Valot, S; Sautour, M; Dalle, F; Muller, C; Borni-Duval, C; Caillard, S; Moulin, B; Pfaff, A W; Razakandrainibe, R; Abou-Bacar, A; Favennec, L; Candolfi, E

    2016-08-02

    Cryptosporidium spp. is a ubiquitous parasite affecting humans as well as domestic and wild vertebrates, causing diarrhea in both immunocompetent and immunocompromised hosts worldwide. Its transmission occurs primarily by the fecal-oral route. In humans, C. parvum and C. hominis are the most prevalent species, whereas immunocompetent and immunocompromised individuals can also be infected by other zoonotic species. Renal transplant patients are prone to develop cryptosporidiosis, which can induce severe and life-threatening diarrhea. We report here a series of nearly concomitant cases of acute symptomatic cryptosporidiosis in three renal transplant patients attending the Strasbourg University Hospital Nephrology Unit. The clinical presentation was persistent diarrhea and acute renal failure. The diagnosis was confirmed by microscopic stool examination using a modified Ziehl-Neelsen staining method and species identification by molecular tools. All patients were treated with nitazoxanide and recovered from diarrhea after 14 days of therapy. Genotypic species identification was not consistent with an epidemic context, thus underlining the need for genotyping to monitor at risk patients.

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

    Science.gov (United States)

    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

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

    Directory of Open Access Journals (Sweden)

    Geisi Ferreira Mariné Oliveira

    2016-04-01

    Full Text Available 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.

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

    Science.gov (United States)

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

    2016-04-01

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

  11. Improved Risk Analysis by Dual Direct Detection of Total and Infectious Cryptosporidium Oocysts on Cell Culture in Combination with Immunofluorescence Assay▿

    Science.gov (United States)

    Lalancette, Cindy; Di Giovanni, George D.; Prévost, Michèle

    2010-01-01

    The inactivation of Cryptosporidium oocysts is a main driver in the selection of water treatment disinfection strategies, and microbial risk analysis provides a sound basis for optimizing water treatment processes. U.S. Environmental Protection Agency method 1622/23 provides an estimate of the total oocyst count; however, it cannot be used directly for risk assessment, as it does not determine the fraction of infectious oocysts. Improved assessment of the risk for designated sources or in treated water requires evaluation of the total number of oocysts and an estimate of their infectivity. We developed a dual direct detection method using differential immunofluorescent staining that allows detection of both oocysts and cell culture infection foci for each sample. Using Cryptosporidium parvum oocysts, various pH levels, proteases, and gastroenteric compounds and substrates were assessed to determine their abilities to enhance the number of infection foci. The results showed that the key trigger for oocyst stimulation was acidification. Addition of a low concentration of d-glucose (50 mM) to the infection media increased rates of infectivity, while a higher dose (300 mM) was inhibitory. The total number of oocysts in each sample was determined by counting the oocysts remaining on a cell monolayer and the oocysts recovered from cell monolayer washes during processing using a simple filtration technique. With the dual direct detection on cell culture with immunofluorescence assay method, it is now possible to determine the numbers of total and infectious oocysts for a given sample in a single analysis. Direct percentages of infectivity are then calculated, which allows more accurate assessments of risk. PMID:19933339

  12. Presence of Cryptosporidium spp. and Giardia duodenalis through drinking water.

    Science.gov (United States)

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

    2008-11-01

    To evaluate the presence of Cryptosporidium spp. and Giardia duodenalis in the influent and final effluent of sixteen drinking water treatment plants located in a hydrographic basin in Galicia (NW Spain) - in which the principal river is recognised as a Site of Community Importance (SCI) - estimate the efficiency of treatment plants in removing these protozoans and determine the species and genotypes of the parasites by means of a molecular assay. All plant samples of influent and final effluent (50-100 l) were examined in the spring, summer, autumn and winter of 2007. A total of 128 samples were analysed by method 1623, developed by US Environmental Protection Agency for isolation and detection of both parasites. To identify the genotypes present the following genes were amplified and sequenced: 18S SSU rRNA (Cryptosporidium spp.) and b-giardina (G. duodenalis). The mean concentrations of parasites in the influent were 0.0-10.5 Cryptosporidium spp. oocysts per litre and 1.0-12.8 of G. duodenalis cysts per litre. In the final treated effluent, the mean concentration of parasites ranged from 0.0-3.0 oocysts per litre and 0.5-4.0 cysts per litre. The distribution of results by season revealed that in all plants, the highest numbers of (oo)cysts were recorded in spring and summer. Cryptosporidium parvum, C. andersoni, C. hominis and assemblages A-I, A-II, E of G. duodenalis were detected. Cryptosporidium spp. and G. duodenalis were consistently found at high concentrations in drinking water destined for human and animal consumption in the hydrographic basin under study, in Galicia (NW Spain). It is important that drinking water treatment authorities rethink the relevance of contamination levels of both parasites in drinking water and develop adequate countermeasures.

  13. First genetic analysis of Cryptosporidium from humans from Tasmania, and identification of a new genotype from a traveller to Bali.

    Science.gov (United States)

    Koehler, Anson V; Whipp, Margaret; Hogg, Geoff; Haydon, Shane R; Stevens, Melita A; Jex, Aaron R; Gasser, Robin B

    2014-09-01

    Little is known about the molecular composition of Cryptosporidium species from humans living in the insular state of Tasmania, Australia. In the present study, we genetically characterized 82 samples of Cryptosporidium from humans following conventional coproscopic testing in a routine, diagnostic laboratory. Using a PCR-coupled single-strand conformation polymorphism (SSCP) technique, targeting portions of the small subunit rRNA (SSU), and 60 kDa glycoprotein (gp60) loci, we identified two species of Cryptosporidium, including C. hominis (subgenotypes IbA10G2, IdA16, IeA12G3T3, and IfA19G1) and C. parvum (IIaA16G1R1 and IIaA18G3), and a new operational taxonomic unit (OTU) that genetically closely resembled C. wrairi. This OTU was further characterized using markers in the actin, Cryptosporidium oocyst wall protein (COWP), and 70 kDa heat shock protein (hsp70) genes. This study provides the first characterization of species and genotypes of Cryptosporidium from Tasmania, and presents clear genetic evidence, using five independent genetic loci, for a new genotype or species of Cryptosporidium in a Tasmanian person with a recent history of travelling to Bali, Indonesia. It would be interesting to undertake detailed molecular-based studies of Cryptosporidium in Indonesia and neighbouring countries, in conjunction with morphological and experimental investigations of new genotypes.

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

    Science.gov (United States)

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

    2014-01-01

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

  15. Prevalence of Cryptosporidium sp. in patients with colorectal cancer.

    Science.gov (United States)

    Sulżyc-Bielicka, Violetta; Kołodziejczyk, Lidia; Jaczewska, Sylwia; Bielicki, Dariusz; Kładny, Józef; Safranow, Krzysztof

    2012-07-01

    Parasitic protozoans of the Cryptosporidium genus are intracellular intestinal parasites of mammals, causing cryptosporidiosis. Clinically, cryptosporidiosis manifests as chronic diarrhoea. Individuals with immune disorders, including those with neoplasms, are at risk of symptomatic invasion. Was the evaluation of Cryptosporidium sp. prevalence in patients with diagnosed colorectal cancer. The studied group encompassed 87 patients with diagnosed colorectal cancer, undergoing surgery at the Department of General and Oncological Surgery, Pomeranian Medical University, in the years 2009-2010. Immunoenzymatic tests for Cryptosporidium sp. on faeces samples were performed with the use of commercial test kit, ProSpecT(®)Cryptosporidium Microplate Assay (Remel Inc). The presence of Cryptosporidium sp. was found in 12.6% of studied patients with colorectal cancer. The performed statistical analysis did not reveal any correlation between Cryptosporidium sp. infection and gender, age, neoplasm advancement stage as per Astler-Coller scale, neoplasm differentiation grade, or neoplastic tumour localisation in relation to the splenic flexure. There was found high prevalence of Cryptosporidium sp. in patients with colorectal cancer. It was comparable to the prevalence reported for patients with immune deficiency.

  16. Identification of Cryptosporidium species and genotypes in Scottish raw and drinking waters during a one-year monitoring period.

    Science.gov (United States)

    Nichols, R A B; Connelly, L; Sullivan, C B; Smith, H V

    2010-09-01

    We analyzed 1,042 Cryptosporidium oocyst-positive slides (456 from raw waters and 586 from drinking waters) of which 55.7% contained 1 or 2 oocysts, to determine species/genotypes present in Scottish waters. Two nested PCR-restriction fragment length polymorphism (RFLP) assays targeting different loci (1 and 2) of the hypervariable region of the 18S rRNA gene were used for species identification, and 62.4% of samples were amplified with at least one of the PCR assays. More samples (577 slides; 48.7% from raw water and 51.3% from drinking water) were amplified at locus 1 than at locus 2 (419 slides; 50.1% from raw water and 49.9% from drinking water). PCR at loci 1 and 2 amplified 45.4% and 31.7% of samples containing 1 or 2 oocysts, respectively. We detected both human-infectious and non-human-infectious species/genotype oocysts in Scottish raw and drinking waters. Cryptosporidium andersoni, Cryptosporidium parvum, and the Cryptosporidium cervine genotype (now Cryptosporidium ubiquitum) were most commonly detected in both raw and drinking waters, with C. ubiquitum being most common in drinking waters (12.5%) followed by C. parvum (4.2%) and C. andersoni (4.0%). Numerous samples (16.6% total; 18.9% from drinking water) contained mixtures of two or more species/genotypes, and we describe strategies for unraveling their identity. Repetitive analysis for discriminating mixtures proved useful, but both template concentration and PCR assay influenced outcomes. Five novel Cryptosporidium spp. (SW1 to SW5) were identified by RFLP/sequencing, and Cryptosporidium sp. SW1 was the fourth most common contaminant of Scottish drinking water (3%).

  17. 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.

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    Samba O Sow

    2016-05-01

    Full Text Available 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

  18. 牧区儿童隐孢子虫感染情况及危险因素的调查%Investigation of the Infection Situation and the Risk Factors in Children of Cryptosporidium

    Institute of Scientific and Technical Information of China (English)

    孙玉荣

    2015-01-01

    目的:通过调查,了解牧区儿童隐孢子虫感染情况,观察牧区饮用水隐孢子虫污染环境情况,及时掌握牧区家养牲畜隐孢子虫感染情况,探讨和分析牧区儿童感染隐孢子虫的危险因素,为制定相关的预防控制策略和措施提供参考数据。方法选择2012年12月-2014年12月6个典型牧区的8所幼儿园1002名儿童粪便、120份牛粪便、120份羊粪便、120份猪粪便用RIDA快速检测法和金胺-酚复染法进行检测。同时向1002名儿童进行问卷调查,分析儿童对隐孢子虫的知晓率以及各类粪便中隐孢子虫检测结果情况。结果儿童对隐孢子虫知晓率为30.03%。1002份儿童粪便样品,有42份样品隐孢子虫卵囊阳性,阳性率为4.19%;120份牛粪便样品,有25份样品隐孢子虫卵囊阳性,阳性率为20.83%;120份羊粪便样品,有14份样品隐孢子虫卵囊阳性,阳性率为12.07%;120份猪粪便样品,有35份样品隐孢子虫卵囊阳性,阳性率为29.17%。结果差异有统计学意义(P<0.05)。结论牧区粪便污染较为严重,且儿童对其知晓程度较低,应加强儿童的健康教育,环境问题不容忽视。%Objective Through investigation, understand the infection status of children in pastoral areas of Cryptosporidium, ob-serve the pastoral areas of drinking water pollution of the environment condition of Cryptosporidium infection, timely grasp of pas-toral cattle Cryptosporidium, discussion and analysis of the risk factors of pastoral children infected with Cryptosporidium, for for-mulating prevention and control strategies and measures to provide relevant reference data. Methods Select from 2012 December to 2014 December 8 kindergartens in 6 typical pastoral 1002 children stool, 120 copies, 120 copies of cow dung manure, sheep, 120 swine feces with a rapid detection method of RIDA and auramine phenol staining method for detection of complex. At the same time, a

  19. Infection Status of Hospitalized Diarrheal Patients with Gastrointestinal Protozoa, Bacteria, and Viruses in the Republic of Korea

    Science.gov (United States)

    Cho, Shin-Hyeong; Lee, Jin-Hee; Lim, Yi-Young; Jeon, Ji-Hye; Yu, Jae-Ran; Kim, Tong-Soo; Lee, Won-Ja; Cho, Seung-Hak; Lee, Deog-Yong; Park, Mi-Seon; Jeong, Hye-Sook; Chen, Doo-Sung; Ji, Yeong-Mi; Kwon, Mi-Hwa

    2010-01-01

    To understand protozoan, viral, and bacterial infections in diarrheal patients, we analyzed positivity and mixed-infection status with 3 protozoans, 4 viruses, and 10 bacteria in hospitalized diarrheal patients during 2004-2006 in the Republic of Korea. A total of 76,652 stool samples were collected from 96 hospitals across the nation. The positivity for protozoa, viruses, and bacteria was 129, 1,759, and 1,797 per 10,000 persons, respectively. Especially, Cryptosporidium parvum was highly mixed-infected with rotavirus among pediatric diarrheal patients (29.5 per 100 C. parvum positive cases), and Entamoeba histolytica was mixed-infected with Clostridium perfringens (10.3 per 100 E. histolytica positive cases) in protozoan-diarrheal patients. Those infected with rotavirus and C. perfringens constituted relatively high proportions among mixed infection cases from January to April. The positivity for rotavirus among viral infection for those aged ≤ 5 years was significantly higher, while C. perfringens among bacterial infection was higher for ≥ 50 years. The information for association of viral and bacterial infections with enteropathogenic protozoa in diarrheal patients may contribute to improvement of care for diarrhea as well as development of control strategies for diarrheal diseases in Korea. PMID:20585526

  20. Infection status of hospitalized diarrheal patients with gastrointestinal protozoa, bacteria, and viruses in the Republic of Korea.

    Science.gov (United States)

    Cheun, Hyeng-Il; Cho, Shin-Hyeong; Lee, Jin-Hee; Lim, Yi-Young; Jeon, Ji-Hye; Yu, Jae-Ran; Kim, Tong-Soo; Lee, Won-Ja; Cho, Seung-Hak; Lee, Deog-Yong; Park, Mi-Seon; Jeong, Hye-Sook; Chen, Doo-Sung; Ji, Yeong-Mi; Kwon, Mi-Hwa

    2010-06-01

    To understand protozoan, viral, and bacterial infections in diarrheal patients, we analyzed positivity and mixed-infection status with 3 protozoans, 4 viruses, and 10 bacteria in hospitalized diarrheal patients during 2004-2006 in the Republic of Korea. A total of 76,652 stool samples were collected from 96 hospitals across the nation. The positivity for protozoa, viruses, and bacteria was 129, 1,759, and 1,797 per 10,000 persons, respectively. Especially, Cryptosporidium parvum was highly mixed-infected with rotavirus among pediatric diarrheal patients (29.5 per 100 C. parvum positive cases), and Entamoeba histolytica was mixed-infected with Clostridium perfringens (10.3 per 100 E. histolytica positive cases) in protozoan-diarrheal patients. Those infected with rotavirus and C. perfringens constituted relatively high proportions among mixed infection cases from January to April. The positivity for rotavirus among viral infection for those aged or = 50 years. The information for association of viral and bacterial infections with enteropathogenic protozoa in diarrheal patients may contribute to improvement of care for diarrhea as well as development of control strategies for diarrheal diseases in Korea.

  1. Prevalence and Genetic Characterization of Cryptosporidium Spp. In Diarrheic Children from Gonbad Kavoos City, Iran

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    Mitra SHARBATKHORI

    2015-10-01

    Full Text Available Background: Cryptosporidium is an intestinal protozean parasite causing water­borne and foodborne outbreaks of diarrheal diseases. The present study was per­formed in order to find prevalence and subtypes of Cryptosporidium among children with diarrhea in Gonbad Kavoos City, Northern Iran.Methods: Diarrheic samples were collected from 547 children. The initial parasitologi­cal diagnosis was made based on detection of oocysts using the modi­fied Ziehl-Neelsen acid-fast staining method. The positive microscopically samples were selected for sequence analysis of partial 60 kDa glycoprotein (gp60 gene. Results: Out of 547 collected samples, 27 (4.94% were positive for Cryptosporid­ium oocysts. Fifteen from 27 positive samples successfully amplified in PCR. Se­quences analysis of gp60 gene in 15 Cryptosporidium isolates revealed that all of them (100% were C. parvum. The results showed three subtypes of IIa subtype family (7 cases including IIaA16G2R1, IIaA17G1R1, IIaA22G3R1 and one subtype of IId subtype family (8 cases. The most common allele was IId A17G1d (53.3%. Conclusion: The predominance of zoonotic subtype families of C. parvum species (IIa, IId in the present study is in concordance with previous studies in Iran and emphasizes the significance of zoonotic transmission of cryptosporidiosis in the country.

  2. SPECIES AND STRAIN-SPECIFIC TYPING OF CRYPTOSPORIDIUM PARASITES IN CLINICAL AND ENVIRONMENTAL SAMPLES

    Science.gov (United States)

    Cryptosporidiosis has recently attracted attention as an emerging water borne and food borne disease as well as an opportunistic infection in HIV infected indivduals. The lack of genetic information, however, has resulted in confusion in the taxonomy of Cryptosporidium parasites ...

  3. Parasitismo por Giardia sp. e Cryptosporidium sp. em Coendou villosus Parasitism by Giardia sp. and Cryptosporidium sp. in Coendou villosus

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    João Fabio Soares

    2008-04-01

    Full Text Available O objetivo deste trabalho foi verificar o possível parasitismo por Giardia sp. e Cryptosporidium sp. em amostras de fezes de ouriço-cacheiro (Coendou villosus. As amostras foram analisadas pelo método de centrífugo-flutuação com sulfato de zinco e apresentaram elevada infecção por cistos de Giardia sp. e por oocistos de Cryptosporidium sp., embora os animais não apresentassem sinal clínico decorrente disso.This research was aimed at verifing the possible parasitism by Giardia sp. and Cryptosporidium sp. in porcupine (Coendou villosus faeces samples. Samples were analyzed by the centrifugal-flotation method with zinc sulphate and showed high infection by cysts of Giardia sp. and by oocysts of Cryptosporidium sp., although the animals did not show any associated clinical sign.

  4. Evolution of mitosome metabolism and invasion-related proteins in Cryptosporidium.

    Science.gov (United States)

    Liu, Shiyou; Roellig, Dawn M; Guo, Yaqiong; Li, Na; Frace, Michael A; Tang, Kevin; Zhang, Longxian; Feng, Yaoyu; Xiao, Lihua

    2016-12-08

    The switch from photosynthetic or predatory to parasitic life strategies by apicomplexans is accompanied with a reductive evolution of genomes and losses of metabolic capabilities. Cryptosporidium is an extreme example of reductive evolution among apicomplexans, with losses of both the mitosome genome and many metabolic pathways. Previous observations on reductive evolution were largely based on comparative studies of various groups of apicomplexans. In this study, we sequenced two divergent Cryptosporidium species and conducted a comparative genomic analysis to infer the reductive evolution of metabolic pathways and differential evolution of invasion-related proteins within the Cryptosporidium lineage. In energy metabolism, Cryptosporidium species differ from each other mostly in mitosome metabolic pathways. Compared with C. parvum and C. hominis, C. andersoni possesses more aerobic metabolism and a conventional electron transport chain, whereas C. ubiquitum has further reductions in ubiquinone and polyisprenoid biosynthesis and has lost both the conventional and alternative electron transport systems. For invasion-associated proteins, similar to C. hominis, a reduction in the number of genes encoding secreted MEDLE and insulinase-like proteins in the subtelomeric regions of chromosomes 5 and 6 was also observed in C. ubiquitum and C. andersoni, whereas mucin-type glycoproteins are highly divergent between the gastric C. andersoni and intestinal Cryptosporidium species. Results of the study suggest that rapidly evolving mitosome metabolism and secreted invasion-related proteins could be involved in tissue tropism and host specificity in Cryptosporidium spp. The finding of progressive reduction in mitosome metabolism among Cryptosporidium species improves our knowledge of organelle evolution within apicomplexans.

  5. Cryptosporidium and Giardia in recreational water in Belgium.

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    Ehsan, Md Amimul; Casaert, Stijn; Levecke, Bruno; Van Rooy, Liesbet; Pelicaen, Joachim; Smis, Anne; De Backer, Joke; Vervaeke, Bart; De Smedt, Sandra; Schoonbaert, Filip; Lammens, Saskia; Warmoes, Thierry; Geurden, Thomas; Claerebout, Edwin

    2015-09-01

    The objective of this study was to investigate the presence of Cryptosporidium and Giardia in different recreational water bodies in Belgium and to estimate the infection risk associated with swimming and other recreational activities. Cryptosporidium oocysts and/or Giardia cysts were detected in three out of 37 swimming pools, seven out of 10 recreational lakes, two out of seven splash parks and four out of 16 water fountains. In the swimming pools no infection risk for Cryptosporidium could be calculated, since oocysts were only detected in filter backwash water. The risk of Giardia infection in the swimming pools varied from 1.13×10(-6) to 2.49×10(-6) per swim per person. In recreational lakes, the infection risk varied from 2.79×10(-5) to 5.74×10(-5) per swim per person for Cryptosporidium and from 7.04×10(-5) to 1.46×10(-4) for Giardia. For other outdoor water recreation activities the estimated infection risk was 5.71×10(-6) for Cryptosporidium and 1.47×10(-5) for Giardia. However, most positive samples in the recreational lakes belonged to species/genotypes that are either animal-specific or predominantly found in animals. No Cryptosporidium was found in splash parks and water fountains, but the presence of Giardia cysts suggests a risk for human infection. The infection risk of Giardia infection during a 3.5-minute visit to a splash park for children equalled 1.68×10(-4).

  6. First molecular characterization of Cryptosporidium from three different points of two main rivers in Kuantan, Malaysia using 18S rRNA gene nested PCR

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    Mohd Aiman Barudin

    2017-09-01

    Full Text Available Objective: To identify 18S ribosomal RNA (18S rRNA partial sequences from three different points, namely, downstream, midstream and upstream of two major rivers in Kuantan, Pahang, Malaysia. Methods: In this study, six river water samples were collected from three different points of both Kuantan River and Balok River. Each water sample was processed and concentrated using continuous flow centrifuge machine and purified using immunomagnetic separation technique. Nested PCR was applied to detect 18S rRNA sequence in those samples after DNA extraction. Genotyping and phylogenetic analysis were carried out based on the gene of 18S rRNA sequence of Cryptosporidium. Results: A total of five samples from six different points of both Kuantan River and Balok River were contaminated with Cryptosporidium. Only river water sample from the upstream point of Kuantan River was negative for Cryptosporidium. In this study, the sequencing results of five samples from both Kuantan River and Balok River belonged to Cryptosporidium parvum. Conclusions: This is the first study of Cryptosporidium parvum genotyping in both Kuantan River and Balok River by using molecular approach nested PCR on 18S rRNA gene.

  7. Molecular evidence of Ureaplasma urealyticum and Ureaplasma parvum colonization in preterm infants during respiratory distress syndrome

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    Germani Rossella

    2006-11-01

    Full Text Available Abstract Background Ureaplasma urealyticum and U. parvum have been associated with respiratory diseases in premature newborns, but their role in the pathogenesis of the respiratory distress syndrome (RDS is unclear. The aim of this study was to detect, using molecular techniques, the role of Mycoplasma spp. and Ureaplasma spp. in respiratory secretion and blood specimens of preterm newborns with or without RDS and to evaluate the prevalence of perinatal U. urealyticum or U. parvum infection. The influence of chemotherapy on the clinical course was also evaluated. Methods Tracheal aspirate or nasopharingeal fluid samples from 50 preterm babies with (24 or without RDS (26 were analysed for detection of U. urealyticum and U. parvum by culture identification assay and PCR. Sequencing analysis of amplicons allowed us to verify the specificity of methods. Clarithromycin (10 mg kg-1 twice a day was administered in ureaplasma-positive patients who presented clinical signs of RDS. Results 15/24 neonates with RDS (p U. urealyticum or U. parvum. Culture identification assay was positive in 5/50 newborns, three of which with RDS. Sequencing analyses confirmed the specificity of these methods. Association of patent ductus arteriosus with ureaplasma colonization was more statistically significant (p = 0.0004 in patients with RDS than in those without RDS. Conclusion Colonization of the lower respiratory tract by Ureaplasma spp. and particularly by U. parvum in preterm newborns was related to RDS. The routine use of molecular methods could be useful to screen candidate babies for etiologic therapy.

  8. Antibody responses to the immunodominant Cryptosporidium gp15 antigen and gp15 polymorphisms in a case-control study of cryptosporidiosis in children in Bangladesh.

    Science.gov (United States)

    Allison, Genève M; Rogers, Kathleen A; Borad, Anoli; Ahmed, Sabeena; Karim, Mohammad Mahbubul; Kane, Anne V; Hibberd, Patricia L; Naumova, Elena N; Calderwood, Stephen B; Ryan, Edward T; Khan, Wasif A; Ward, Honorine D

    2011-07-01

    Although Cryptospridium hominis is the dominant Cryptosporidium species infecting humans, immune responses to cognate antigens in C. hominis-infected persons have not been reported. We investigated antibody responses to the immunodominant gp15 antigen from C. hominis and C. parvum, in C. hominis-infected Bangladeshi children less than five years of age with diarrhea (cases) and uninfected children with diarrhea (controls). We also investigated polymorphisms in the C. hominis gp15 sequence from cases. Serum IgG responses to gp15 from both species were significantly greater in cases than controls. In spite of polymorphisms in the gp15 sequence, there was a significant correlation between antibody levels to gp15 from both species, indicating cross-reactivity to conserved epitopes. Cases with acute diarrhea had a significantly greater serum IgA response to gp15 compared with those with persistent diarrhea, suggesting that this response may be associated with protection from prolonged disease. These findings support further investigation of gp15 as a vaccine candidate.

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

    Science.gov (United States)

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

    2013-06-01

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

  10. Molecular epidemiology of Cryptosporidium and Giardia in cattle

    Science.gov (United States)

    Cryptosporidium spp. and Giardia duodenalis are enteric protozoan parasites that infect a wide range of vertebrate hosts including humans. Infections with both parasites are known as one of the most common causes of diarrhea in humans and livestock. The epidemiology of cryptosporidiosis and giardias...

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

    Science.gov (United States)

    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.

  12. Parasitic infections in HIV infected individuals: Diagnostic & therapeutic challenges

    Science.gov (United States)

    Nissapatorn, Veeranoot; Sawangjaroen, Nongyao

    2011-01-01

    After 30 years of the human immunodeficiency virus (HIV) epidemic, parasites have been one of the most common opportunistic infections (OIs) and one of the most frequent causes of morbidity and mortality associated with HIV-infected patients. Due to severe immunosuppression, enteric parasitic pathogens in general are emerging and are OIs capable of causing diarrhoeal disease associated with HIV. Of these, Cryptosporidium parvum and Isospora belli are the two most common intestinal protozoan parasites and pose a public health problem in acquired immunodeficiency syndrome (AIDS) patients. These are the only two enteric protozoan parasites that remain in the case definition of AIDS till today. Leismaniasis, strongyloidiasis and toxoplasmosis are the three main opportunistic causes of systemic involvements reported in HIV-infected patients. Of these, toxoplasmosis is the most important parasitic infection associated with the central nervous system. Due to its complexity in nature, toxoplasmosis is the only parasitic disease capable of not only causing focal but also disseminated forms and it has been included in AIDS-defining illnesses (ADI) ever since. With the introduction of highly active anti-retroviral therapy (HAART), cryptosporidiosis, leishmaniasis, schistosomiasis, strongyloidiasis, and toxoplasmosis are among parasitic diseases reported in association with immune reconstitution inflammatory syndrome (IRIS). This review addresses various aspects of parasitic infections in term of clinical, diagnostic and therapeutic challenges associated with HIV-infection. PMID:22310820

  13. Experimental transmission of Cryptosporidium oocyst isolates from mammals, birds and reptiles to captive snakes.

    Science.gov (United States)

    Graczyk, T K; Cranfield, M R

    1998-01-01

    Groups of four to five, 3-month-old rat snakes (Elaphe obsoleta) were separately gastrically inoculated with 2.0 x 10(6) viable oocysts of Cryptosporidium muris (mice and calves), C. muris-like (Bactrian camels), C. wrairi (guinea pigs), C. baileyi (chickens), C. meleagridis (turkeys), Cryptosporidium sp. (turtles, tortoises, chameleons and lizards) and C. serpentis from clinically (fatal case) and subclinically infected snakes. None of the snakes inoculated with oocysts originating from homothermous vertebrates developed infection as determined by histology and serology, whereas all snakes challenged with reptilian oocyst isolates were infected with Cryptosporidium on weeks 6 and 10 post-inoculation (PI). One week 10 PI, the snakes displayed mild to serve, multifocal to widespread, thinning and disorganization of gastric epithelium and nine out of twelve snakes infected by oocysts originating from reptiles other than snakes displayed severe gastric hyperplasia. Three out of ten snakes infected oocysts originating from snakes had ELISA-detectable Cryptosporidium-specific antibody (Ab) titers on week 6 PI; all snakes were Cryptosporidium-seroconverted on week 10 PI and their serum Ab titer significantly increased. The study demonstrated that Cryptosporidium infections in snakes maintained on the diet of rodents or birds cannot be initiated via ingestion of an infected food item; however, snakes can void ingested oocysts. Lack of host specificity among reptiles to this pathogen, demonstrated for the first time in the present study, indicates that snake-attributed C. serpentis is not distinct from Cryptosporidium sp. infecting reptiles other than snakes, and that clinical manifestations and virulence of Cryptosporidium in snakes in modulated by the species of the host. Housing of snakes with other reptiles can enhance transmission of Cryptosporidium to snakes, and therefore should be avoided.

  14. Opportunistic intestinal protozoal infections in immunocompromised children.

    Science.gov (United States)

    Noureldin, M S; Shaltout, A A; El Hamshary, E M; Ali, M E

    1999-01-01

    Hundred immunocompromised children and 100 house contact controls were chosen. Patients included: 52 nephrotic syndrome children receiving corticosteroids for more than one month (age 5.28 +/- 2.32 years), 14 protein-calorie malnutrition (PCM) patients (8 cases of marasmus aged 6 +/- 2.27 months and 6 cases of marasmic kwashiorkor aged 1.39 +/- 0.88 years) and 34 lymphomas patients (22 cases of Hodgkin's disease and 12 cases of non-Hodgkin's lymphoma; age 4.5 +/- 3.54 years). Examination of concentrated stool was done using iodine stain of fresh mounts and modified Ziehl-Neelsen (cold acid-fast) to fixed smears. T-cell subsets were counted after staining with mouse monoclonal antibodies against CD4 and CD8 labeled with fluorescein. Both nephrotic syndrome and lymphomas groups showed affection of cellular immunity in the form of significant decrease in T-helper and H/S ratio and significant increase in suppressor T-cell subsets. Giardia lamblia, Entamoeba histolytica, Cryptosporidium parvum and Blastocystis hominis were the most frequent in patients group and were significantly more prevalent among patients than controls. No significant difference in the prevalence of Entamoeba coli and Chylomastix mesnili between the two groups. C. parvum infection were strictly confined to groups with T-cell subsets abnormalities i.e. nephrotic syndrome and lymphomas groups.

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

    Science.gov (United States)

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

    2016-10-01

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

  16. Artemisinin and its derivatives in treating protozoan infections beyond malaria.

    Science.gov (United States)

    Loo, Cecilia Shi Ni; Lam, Nelson Siu Kei; Yu, Deying; Su, Xin-Zhuan; Lu, Fangli

    2017-03-01

    Parasitic protozoan diseases continue to rank among the world's greatest global health problems, which are also common among poor populations. Currently available drugs for treatment present drawbacks, urging the need for more effective, safer, and cheaper drugs. Artemisinin (ART) and its derivatives are some of the most important classes of antimalarial agents originally derived from Artemisia annua L. However, besides the outstanding antimalarial and antischistosomal activities, ART and its derivatives also possess activities against other parasitic protozoa. In this paper we review the activities of ART and its derivatives against protozoan parasites in vitro and in vivo, including Leishmania spp., Trypanosoma spp., Toxoplasma gondii, Neospora caninum, Eimeria tenella, Acanthamoeba castellanii, Naegleria fowleri, Cryptosporidium parvum, Giardia lamblia, and Babesia spp. We conclude that ART and its derivatives may be good alternatives for treating other non-malarial protozoan infections in developing countries, although more studies are necessary before they can be applied clinically.

  17. Multilocus typing and population structure of Cryptosporidium from children in Zaragoza, Spain.

    Science.gov (United States)

    Ramo, Ana; Quílez, Joaquín; Vergara-Castiblanco, Claudia; Monteagudo, Luis; Del Cacho, Emilio; Clavel, Antonio

    2015-04-01

    A multilocus typing approach with eight variable-number tandem-repeat (VNTR) loci and the GP60 gene was used to analyze the inter- and intra-species variation of 44 Cryptosporidium isolates from pediatric patients in Zaragoza city (NE, Spain). Restriction and sequence analyses of the SSU rRNA gene revealed that Cryptosporidium transmission is mostly anthroponotic in this area, with the predominance of Cryptosporidium hominis (n: 41) over Cryptosporidium parvum (n: 3). GP60 subtyping showed limited genetic diversity and four subtypes were identified, including IbA10G2 (n: 35), IaA24R3 (n: 6), IIaA15G1R1 (n: 1) and IIaA15G2R1 (n: 2). Five out of eight VNTR loci showed a discriminatory power higher than the GP60 gene, although each locus had a predominant allele exhibited by more than 50% of isolates. All but four alleles were associated to either C. hominis or C. parvum and linked alleles at different loci were found. Multilocus typing substantially increased the discriminatory power (Hunter-Gaston index: 0.807, 95% CI, 0.683-0.926) and revealed that genetic diversity is much higher than that reported by GP60 sequencing, since 17 multilocus subtypes (MLTs) were identified. Nearly half of the specimens were allocated to a single major MLT. However, no more than three specimens were allocated to each of the remaining MLTs. Both phylogenetic and population analyses revealed a population clustering of C. hominis according to the GP60 subtype, which indicates the robustness of this marker to differentiate genetic subpopulations. Subpopulations had an overall clonal genetic structure, although traces of genetic flow between them were also observed.

  18. Effect of Ureaplasma parvum co-incubation on Chlamydia trachomatis maturation in human epithelial HeLa cells treated with interferon-γ.

    Science.gov (United States)

    Yamazaki, Tomohiro; Matsuo, Junji; Nakamura, Shinji; Oguri, Satoshi; Yamaguchi, Hiroyuki

    2014-08-01

    Chlamydia trachomatis is an obligate intracellular bacterium that causes a sexually transmitted disease. Ureaplasma parvum is commensal in the human genital tract, with a minimal contribution to urogenital infection. We have recently found that U. parvum has a significant effect on the presence of C. trachomatis in the genital tract of healthy women. We therefore assessed the effect of U. parvum co-incubation on C. trachomatis maturation from reticulate bodies (RBs) to elementary bodies (EBs) in HeLa cells in the absence or presence of interferon (IFN)-γ, which is a critical host defense factor. IFN-γ stimulation of viable U. parvum significantly prompted chlamydial growth with an increase in infectious particles, EBs, in HeLa cells. IFN-γ treatment of killed U. parvum had a similar effect on C. trachomatis maturation in HeLa cells. There was no change in expression of indoleamine 2,3-dioxygenase (IDO) in cultures of viable or killed U. parvum. We concluded that U. parvum co-incubation by IFN-γ helped C. trachomatis to mature from RBs to EBs in HeLa cells, independent of IDO expression. This suggests a novel survival strategy of C. trachomatis against IFN-γ exposure, prompting secondary infection of the genital mucosa, with possible clinical implications.

  19. Development of a Cryptosporidium oocyst assay using an automated fiber optic-based biosensor

    Directory of Open Access Journals (Sweden)

    Kramer Marianne F

    2007-10-01

    Full Text Available Abstract An intestinal protozoan parasite, Cryptosporidium parvum, is a major cause of waterborne gastrointestinal disease worldwide. Detection of Cryptosporidium oocysts in potable water is a high priority for the water treatment industry to reduce potential outbreaks among the consumer populace. Anti-Cryptosporidium oocyst polyclonal and monoclonal antibodies were tested as capture and detection reagents for use in a fiber optic biosensor assay for the detection of Cryptosporidium oocysts. Antibodies were validated using enzyme-linked immunosorbent assays, flow cytometry, Western blotting and fluorescent microscopy. Oocysts could be detected at a concentration of 105 oocysts/ml when the polyclonal antibodies were used as the capture and detection reagents. When oocysts were boiled prior to detection, a ten-fold increase in sensitivity was achieved using the polyclonal antibody. Western blotting and immunofluorescence revealed that both the monoclonal and polyclonal antibodies recognize a large (>300 kDa molecular weight mucin-like antigen present on the surface of the oocyst wall. The polyclonal antibody also reacted with a small (105 kDa molecular weight antigen that was present in boiled samples of oocysts. Preliminary steps to design an in-line biosensor assay system have shown that oocysts would have to be concentrated from water samples and heat treated to allow detection by a biosensor assay.

  20. Elevation and vegetation determine Cryptosporidium oocyst shedding by yellow-bellied marmots (Marmota flaviventris in the Sierra Nevada Mountains

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    Diego Montecino-Latorre

    2015-08-01

    Full Text Available Wildlife are increasingly recognized as important biological reservoirs of zoonotic species of Cryptosporidium that might contaminate water and cause human exposure to this protozoal parasite. The habitat range of the yellow-bellied marmot (Marmota flaviventris overlaps extensively with the watershed boundaries of municipal water supplies for California communities along the foothills of the Sierra Nevada. We conducted a cross-sectional epidemiological study to estimate the fecal shedding of Cryptosporidium oocysts by yellow-bellied marmots and to quantify the environmental loading rate and determine risk factors for Cryptosporidium fecal shedding in this montane wildlife species. The observed proportion of Cryptosporidium positive fecal samples was 14.7% (33/224, positive number relative to total number samples and the environmental loading rate was estimated to be 10,693 oocysts animal-1 day-1. Fecal shedding was associated with the elevation and vegetation status of their habitat. Based on a portion of the 18s rRNA gene sequence of 2 isolates, the Cryptosporidium found in Marmota flaviventris were 99.88%–100% match to multiple isolates of C. parvum in the GenBank.

  1. Prevalence rates of Giardia and Cryptosporidium among diarrheic patients in the Philippines.

    Science.gov (United States)

    Natividad, Filipinas F; Buerano, Corazon C; Lago, Catherine B; Mapua, Cynthia A; de Guzman, Blanquita B; Seraspe, Ebonia B; Samentar, Lorena P; Endo, Takuro

    2008-11-01

    The prevalence of Giardia and Cryptosporidium among 3,456 diarrheic patients corrected from May 2004 to May 2005 in the Philippines was determined. Of 133 (3.8%) positive samples, 69 (2.0%) were positive for Giardia and 67 (1.9%) for Cryptosporidium. Three samples had co-infection with Giardia and Cryptosporidium. Luzon had the highest positive samples (5.0%) followed by Mindanao (4.9%), then Visayas (2.2%). Giardia was most prevalent in Mindanao (3.6%) while Cryptosporidium was most prevalent in Luzon (3.1%). The prevalence of Giardia (2.0%) among pediatric patients (0-18 years) did not significantly differ from that (1.9%) among adults (> 18 years old). However, for Cryptosporidium, the prevalence (2.9%) among pediatric patients was significantly higher compared to that (0.2%) among adult patients. In the pediatric population, the highest percentage of patients with Giardia was the 5-9 year old age group, while that of Cryptosporidium was in the 0-4 year old group. The prevalence of Giardia, but not Cryptosporidium, was significantly higher in male than female adults. Seasonality had a distinct peak in September with Cryptosporidium more prevalent in the rainy (2.6%) than dry season (0.9%).

  2. The first report of Cryptosporidium andersoni in horses with diarrhea and multilocus subtype analysis.

    Science.gov (United States)

    Liu, Aiqin; Zhang, Jia; Zhao, Jingmin; Zhao, Wei; Wang, Rongjun; Zhang, Longxian

    2015-09-22

    Horses interact with humans in a wide variety of sport competitions and non-competitive recreational pursuits as well as in working activities. Cryptosporidium spp are one of the most important zoonotic pathogens causing diarrhea of humans and animals. The reports of Cryptosporidium in horses and the findings of zoonotic Cryptosporidium species/genotypes show a necessity to carry out molecular identification of Cryptosporidium in horses, especially in diarrheic ones. The aim of the present study was to understand Cryptosporidium infection and species/genotypes in diarrheic horses, and to trace the source of infection of horse-derived Cryptosporidium isolates at a subtype level. Fecal specimens of 29 diarrheic adult horses were collected in Taikang County in northeastern China's Heilongjiang Province. Cryptosporidium oocysts were concentrated by Sheather's sugar flotation technique, and then examined by a bright-field microscope. Meanwhile, all the specimens were subjected to PCR amplification of the small subunit (SSU) rRNA gene of Cryptosporidium. C. andersoni isolates were further subtyped by multilocus sequence typing (MLST) at the four microsatellite/minisatellite loci (MS1, MS2, MS3 and MS16). One and two Cryptosporidium-positive isolates were obtained in horses by microscopy and by PCR, respectively. The two C. andersoni isolates were identified by sequencing of the SSU rRNA gene of Cryptosporidium. Both of them were identical to each other at the MS1, MS2, MS3 and MS16 loci, and MLST subtype A4,A4,A4,A1 was found here. This is the first report of C. andersoni in horses. The fact that the MLST subtype A4,A4,A4,A1 was reported in cattle suggests a large possibility of transmission of C. andersoni between cattle and horses.

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

    Science.gov (United States)

    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%)(Plamblia 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.

  4. The prevalence of Cryptosporidium spp. oocysts in wild mammals in the Kruger National Park, South Africa.

    Science.gov (United States)

    Abu Samra, Nada; Jori, Ferran; Samie, Amidou; Thompson, Peter

    2011-01-10

    This study determined the prevalence of Cryptosporidium spp. oocysts in faecal samples from elephant (Loxodonta africana), buffalo (Syncerus caffer) and impala (Aepyceros melampus) in the Kruger National Park (KNP) and an adjacent game reserve in South Africa. Two of the study areas were in close proximity to rural communities on the western KNP boundary and the third study area was located in the centre of the KNP. Fresh stool samples (n=445) were collected and tested using an immunofluorescent antibody test (IFA) for Cryptosporidium parvum. A total of 278 of these were randomly selected (approximately 90 samples per wildlife species) and tested with the modified Ziehl Neelsen staining technique (ZN) for Cryptosporidium spp. The prevalence of Cryptosporidium spp. was highest in elephants (25.8% [95% confidence interval: 17.3, 35.9]), compared to buffalo (5.5% [1.8, 12.4]) and impala (4.3% [1.2, 10.5]). C. parvum showed similar patterns, being most prevalent in elephants (4.2% [1.5, 8.8]), compared to buffalo (1.4% [0.2, 5.1]) and impala (1.9% [0.4, 5.3]). 29 samples, including ZN positive and IFA positive samples, were retested using a real time PCR (rtPCR) technique. Of the 28 ZN-positive samples, 14 (50%) were positive with rtPCR and of the 9 IFA-positive samples 6 (67%) were confirmed positive by rtPCR. The prevalence of Cryptosporidium oocysts was significantly higher in both of the two study areas adjacent to the western KNP boundary compared to the area in the centre of the KNP (OR=3.2 [1.2, 9.0]; P=0.024). Our study demonstrates for the first time the presence of Cryptosporidium spp. in wildlife in South Africa. The transmission of this parasite between wildlife, domestic animals and humans is a plausible hypothesis and represents a potential risk for immunodeficient human populations.

  5. Genomic Variation in IbA10G2 and Other Patient-Derived Cryptosporidium hominis Subtypes.

    Science.gov (United States)

    Sikora, Per; Andersson, Sofia; Winiecka-Krusnell, Jadwiga; Hallström, Björn; Alsmark, Cecilia; Troell, Karin; Beser, Jessica; Arrighi, Romanico B G

    2017-03-01

    In order to improve genotyping and epidemiological analysis of Cryptosporidium spp., genomic data need to be generated directly from a broad range of clinical specimens. Utilizing a robust method that we developed for the purification and generation of amplified target DNA, we present its application for the successful isolation and whole-genome sequencing of 14 different Cryptosporidium hominis patient specimens. Six isolates of subtype IbA10G2 were analyzed together with a single representative each of 8 other subtypes: IaA20R3, IaA23R3, IbA9G3, IbA13G3, IdA14, IeA11G3T3, IfA12G1, and IkA18G1. Parasite burden was measured over a range of more than 2 orders of magnitude for all samples, while the genomes were sequenced to mean depths of between 17× and 490× coverage. Sequence homology-based functional annotation identified several genes of interest, including the gene encoding Cryptosporidium oocyst wall protein 9 (COWP9), which presented a predicted loss-of-function mutation in all the sequence subtypes, except for that seen with IbA10G2, which has a sequence identical to the Cryptosporidium parvum reference Iowa II sequence. Furthermore, phylogenetic analysis showed that all the IbA10G2 genomes form a monophyletic clade in the C. hominis tree as expected and yet display some heterogeneity within the IbA10G2 subtype. The current report validates the aforementioned method for isolating and sequencing Cryptosporidium directly from clinical stool samples. In addition, the analysis demonstrates the potential in mining data generated from sequencing multiple whole genomes of Cryptosporidium from human fecal samples, while alluding to the potential for a higher degree of genotyping within Cryptosporidium epidemiology.

  6. Intestinal parasites infection among immunocompromised patients in Riyadh, Saudi Arabia.

    Science.gov (United States)

    Al-Megrin, Wafa A I

    2010-04-15

    This study was conducted to determine the prevalence of intestinal parasites among immunocompromised patients in Riyadh, Saudi Arabia. Fecal samples were collected from 136 immunocompromised patients (52 females and 84 males), aged 2 to 69 years. Specimens were fixed with 10% formalin for 30 min (2-3 g fecal(-1)) and then concentrated by a formalin-ether sedimentation technique. Samples were examined as wet saline mounts and in iodine preparation for detection of protozoan oocysts, cysts, helminthic eggs and larvae. Permanent stained smears were performed for intestinal coccidian parasites by the modified Ziehl-Neelsen technique and the modified trichrome stain. Intestinal parasites were detected in 54 (39.7%) most of them (25.7%) were with diarrhea. More specifically the following parasites were detected Cryptosporidium parvum 11(8.1%), Giardia lamblia 9 (6.6%), Cyclospora cayetanensis 8 (5.9%), Blastocystis hominis 7 (5.2%), Entamoeba histolytica 7 (5.2%), Entamoeba coli 5 (3.7%), Strongyloides stercoralis 3 (2.2%), Ascaris lumbricoides 1 (0.7%), Hymenolepis nana 1 (0.7%), Dicrocoelium dendriticum 1 (0.7%) and Hook worm 1 (0.7%). The most common parasite associated with diarrhea in patients, was C. parvum (p < 0.001), followed by G. lamblia, C. cayetanensis and E. histolytica. However, differences between different age groups were statistically non-significant. Similarly, no significant difference in the infection rate were found between male and female patients.

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

    Science.gov (United States)

    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

  8. Origin of a major infectious disease in vertebrates: The timing of Cryptosporidium evolution and its hosts.

    Science.gov (United States)

    Garcia-R, Juan C; Hayman, David T S

    2016-11-01

    Protozoan parasites of the genus Cryptosporidium infect all vertebrate groups and display some host specificity in their infections. It is therefore possible to assume that Cryptosporidium parasites evolved intimately aside with vertebrate lineages. Here we propose a scenario of Cryptosporidium-Vertebrata coevolution testing the hypothesis that the origin of Cryptosporidium parasites follows that of the origin of modern vertebrates. We use calibrated molecular clocks and cophylogeny analyses to provide and compare age estimates and patterns of association between these clades. Our study provides strong support for the evolution of parasitism of Cryptosporidium with the rise of the vertebrates about 600 million years ago (Mya). Interestingly, periods of increased diversification in Cryptosporidium coincides with diversification of crown mammalian and avian orders after the Cretaceous-Palaeogene (K-Pg) boundary, suggesting that adaptive radiation to new mammalian and avian hosts triggered the diversification of this parasite lineage. Despite evidence for ongoing host shifts we also found significant correlation between protozoan parasites and vertebrate hosts trees in the cophylogenetic analysis. These results help us to understand the underlying macroevolutionary mechanisms driving evolution in Cryptosporidium and may have important implications for the ecology, dynamics and epidemiology of cryptosporidiosis disease in humans and other animals.

  9. Cryptosporidium baileyi infection associated with an outbreak of ocular and respiratory disease in otus owls (Otus scops) in a rehabilitation centre.

    Science.gov (United States)

    Molina-Lopez, Rafael A; Ramis, Antoni; Martin-Vazquez, Samuel; Gomez-Couso, Hipolito; Ares-Mazas, Elvira; Caccio, Simone Mario; Leiva, Marta; Darwich, Laila

    2010-06-01

    Cryptosporidiosis has been reported in more than 30 avian species worldwide. Although some cases of cryptosporidiosis have been described in captive birds of prey in the order Falconiformes, to date there have been no reports of the disease in wild raptors. Here we describe for first time an ocular and respiratory disease associated with Cryptosporidium baileyi in wild scops owl (Otus scops, order: Strigiformes). Sixteen otus owl fledglings born in the wild during the summer of 2008 were admitted to the Torreferrussa Wildlife Rehabilitation Centre (Catalonia, northern Spain) in July and August of the same year. In the middle of September, blepharoedema, conjunctival hyperaemia and mucopurulent ocular discharge were diagnosed unilaterally in 75% (12/16) of the birds and bilaterally in 25% (4/16). Moreover, five birds (31%) developed diffuse epithelial corneal oedema, one owl (6%) displayed mild anterior exudative uveitis and another developed rhinitis (6%). Two birds were euthanized because of the severity of disease. The histopathology demonstrated cryptosporidia-like structures in the conjunctival cells and in the nasal respiratory epithelium of one owl. Cryptosporidium spp. oocysts (6.5 to 7.0 x 5.0 to 5.5 microm) were identified by immunofluorescent antibody test (IFAT) in histological sections from eyelids, trachea and respiratory sinuses and in swab samples from the glottis, choanal slit and conjunctival sac. Polymerase chain reaction and DNA sequence analysis confirmed the presence of C. baileyi. Birds were treated orally with azithromycin (40 mg/kg) once a day for 15 days, and by the end of the treatment all owls tested negative for the parasites, by IFAT, and did not display further signs of disease.

  10. Development of a PCR Diagnostic Kit for Cryptosporidium andersoni in Dairy Cow

    Institute of Scientific and Technical Information of China (English)

    ZHOU Rong-qiong; LI Guo-qing; XIAO Shu-min; LI Wei-hua

    2007-01-01

    Cryptosporidiosis is an important zoonosis caused by the Cryptosporidium species. To develop a PCR diagnostic kit for molecular detection and differential diagnosis of Cryptosporidiurn spp., a portion of ITS-1 sequence of Cryptosporidium. Andersoni was chosen as the target DNA for designing the species-specific primers (ZRQF/ZR). The kit components were determined after the PCR amplification conditions were serially optimized. A series of tests were conducted in the specificity, sensitivity, stability, reproducibility, and stored period of the kit, respectively. The results showed that only C. Andersoni were amplified specific band of about 500 bp, while Cryptosporidium. Parvum, Cryptosporidium. Baileyi, Eimeria sp of dairy cow, Toxoplasma gondii, Eimeria sp of pig, Ascaris suum, Cyclospora sp, and E. Coli could not be amplified. 254 oocysts of C. Andersoni was the lowest number that could be detected using the kit. The kit worked well after being stored at room temperature, 4 and -20℃ for nine months. Fecal specimens, which were collected from a total of 243 calves on four different dairy farms in Guangdong Province, China, and one dairy farm in Henan Province, China, were examined using the kit; the positive rate of the kit was 2-13% higher than that of the routine methods. The results indicated that the kit can detect fecal samples faster, more sensitively, and conveniently, and can provide a useful tool for the identification and differentiation of C. Andersoni from the other Cryptosporidium species; it also has implications for further studies on molecular epidemiology and differential diagnostics of cryptosporidiosis in animals.

  11. PROPHYLACTIC EFFECT OF BOVINE ANTI-CRYPTOSPORIDIUM HYPERIMMUNE COLOSTRUM IN HEALTHY VOLUNTEERS CHALLENGED WITH CRYPTOSPORIDIUM PARVUM. (R824759)

    Science.gov (United States)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  12. Ocurrence of Cryptosporidium spp. in Amazonian manatees (Trichechus inunguis, Natterer, 1883

    Directory of Open Access Journals (Sweden)

    Maria Aparecida da Glória Faustino

    2007-09-01

    Full Text Available The apicomplexa protozoa Cryptosporidium infects several mammals, including terrestrial and aquatic species. In the epidemiology of this infection, the ingestion of water and/or food contamined with oocysts comprises the main mechanism of transmission to susceptible animals. Among the Sirenians, the occurrence of this coccidium has been reported in dugongs (Dugong dugon and Antillean manatee (Trichechus manatus manatus. The present study was conducted with the aim of verifying the occurrence of Cryptosporidium spp. in Amazonian manatee. For this purpose, fecal samples were collected from ten free-ranging Amazonian manatees, two specimens in captivity, and 103 supernatants fecal samples. The samples were processed by the sedimentation method in formol-ether and Kinyoun stain technique for the presence of Cryptosporidium spp.. The positive samples were then submitted to Direct Immunoflorescence Test. The results showed 4.34% (05/115 of positive samples. This is the first report of Cryptosporidium spp. in the Amazonian manatee.

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

    Science.gov (United States)

    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.

  14. Giardia and Cryptosporidium in Pristine Protected Catchments in Central Eastern Australia%澳大利亚中东部原始汇水保护区里的表吮贾第虫Giardia和隐小孢子虫Crptosporidium

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    @@ 我们首次证实在澳大利亚中东部没有都市和农业污染的原始汇水保护区的偏远小溪里存在着致病的原生动物表吮贾第虫(肠兰伯氏鞭毛虫)Giardia lamblia和隐小孢子虫Cryptosporidium parvum.

  15. Long-term monitoring of microsporidia, Cryptosporidium and Giardia infections in western Lowland Gorillas (Gorilla gorilla gorilla at different stages of habituation in Dzanga Sangha Protected Areas, Central African Republic.

    Directory of Open Access Journals (Sweden)

    Bohumil Sak

    Full Text Available BACKGROUND: Infectious diseases pose one of the greatest threats to endangered species, and a risk of gastrointestinal parasite transmission from humans to wildlife has always been considered as a major concern of tourism. Increased anthropogenic impact on primate populations may result in general changes in communities of their parasites, and also in a direct exchange of parasites between humans and primates. AIMS: To evaluate the impact of close contact with humans on the occurrence of potentially zoonotic protists in great apes, we conducted a long-term monitoring of microsporidia, Cryptosporidium and Giardia infections in western lowland gorillas at different stages of the habituation process, humans, and other wildlife in Dzanga-Sangha Protected Areas in the Central African Republic. RESULTS: We detected Encephalitozoon cuniculi genotypes I and II (7.5%, Enterocytozoon bieneusi genotype D and three novel genotypes (gorilla 1-3 (4.0%, Giardia intestinalis subgroup A II (2.0% and Cryptosporidium bovis (0.5% in gorillas, whereas in humans we found only G. intestinalis subgroup A II (2.1%. In other wild and domestic animals we recorded E. cuniculi genotypes I and II (2.1%, G. intestinalis assemblage E (0.5% and C. muris TS03 (0.5%. CONCLUSION: Due to the non-specificity of E. cuniculi genotypes we conclude that detection of the exact source of E. cuniculi infection is problematic. As Giardia intestinalis was recorded primarily in gorilla groups with closer human contact, we suggest that human-gorilla transmission has occurred. We call attention to a potentially negative impact of habituation on selected pathogens which might occur as a result of the more frequent presence of humans in the vicinity of both gorillas under habituation and habituated gorillas, rather than as a consequence of the close contact with humans, which might be a more traditional assumption. We encourage to observe the sections concerning hygiene from the IUCN best

  16. Developing risk models of Cryptosporidium transport in soils from vegetated, tilted soilbox experiments.

    Science.gov (United States)

    Harter, Thomas; Atwill, Edward R; Hou, Lingling; Karle, Betsy M; Tate, Kenneth W

    2008-01-01

    Transport of Cryptosporidium parvum through macroporous soils is poorly understood yet critical for assessing the risk of groundwater contamination. We developed a conceptual model of the physics of flow and transport in packed, tilted, and vegetated soilboxes during and immediately after a simulated rainfall event and applied it to 54 experiments implemented with different soils, slopes, and rainfall rates. Using a parsimonious inverse modeling procedure, we show that a significant amount of subsurface outflow from the soilboxes is due to macropore flow. The effective hydraulic properties of the macropore space were obtained by calibration of a simple two-domain flow and transport model that accounts for coupled flow in the matrix and in the macropores of the soils. Using linear mixed-effects analysis, macropore hydraulic properties and oocyst attenuation were shown to be associated with soil bulk density and rainfall rate. Macropore flow was shown to be responsible for bromide and C. parvum transport through the soil into the underlying pore space observed during the 4-h experiments. We confirmed this finding by conducting a pair of saturated soil column studies under homogeneously repacked conditions with no macropores in which no C. parvum transport was observed in the effluent. The linear mixed-effects and logistic regression models developed from the soilbox experiments provide a basis for estimating macropore hydraulic properties and the risk of C. parvum transport through shallow soils from bulk density, precipitation, and total shallow subsurface flow rate. The risk assessment is consistent with the reported occurrence of oocysts in springs or groundwater from fractured or karstic rocks protected only by shallow overlying soils.

  17. Occurrence of Cryptosporidium spp. and Giardia spp. in a public water-treatment system, Paraná, Southern Brazil

    Directory of Open Access Journals (Sweden)

    Jonatas Campos Almeida

    Full Text Available The purpose of this study was to investigate the occurrence of Cryptosporidium spp. and Giardia spp. in a public water-treatment system. Samples of raw and treated water were collected and concentrated using the membrane filtration technique. Direct Immunofluorescence Test was performed on the samples. DNA extraction using a commercial kit was performed and the DNA extracted was submitted to a nested-PCR reaction (n-PCR and sequencing. In the immunofluorescence, 2/24 (8.33% samples of raw water were positive for Giardia spp.. In n-PCR and sequencing, 2/24 (8.33% samples of raw water were positive for Giardia spp., and 2/24 (8.33% samples were positive for Cryptosporidium spp.. The sequencing showed Cryptosporidium parvum and Giardia duodenalis DNA. In raw water, there was moderate correlation among turbidity, color and Cryptosporidium spp. and between turbidity and Giardia spp.. The presence of these protozoans in the water indicates the need for monitoring for water-treatment companies.

  18. [Protozoan infection causes diarrhea in calves].

    Science.gov (United States)

    Geurden, T; Claerebout, E; Vercruysse, J

    2005-12-01

    The role of protozoan parasites in the etiology of diarrhea in calves is highlighted with emphasis on correct diagnosis. In neonatal calves, Cryptosporidium parvum is isolated in more than 44% of the faeces of diarrhetic calves. In calves older than one month, both Eimeria bovis and E. zuernii, and Giardia duodenalis are associated with diarrhea and poor growth. Clinical diagnosis has to be confirmed by examination of host faecal material. Both for C. parvum and G. duodenalis immunological assays are available. Control measures must aim to reduce or prevent oocyst or cyst transmission, by combining management measures, desinfection and chemotherapeutic treatment.

  19. Cryptosporidium cuniculus and Giardia duodenalis in rabbits: genetic diversity and possible zoonotic transmission.

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    Weizhe Zhang

    Full Text Available BACKGROUND: Cryptosporidium and Giardia are the two important zoonotic pathogens causing diarrhea of humans and animals worldwide. Considering the human cryptosporidiosis outbreak and sporadic cases caused by C. cuniculus, the important public health significance of G. duodenalis and little obtained information regarding rabbit infected with Cryptosporidium and Giardia in China, the aim of this study is to determine the prevalence and molecularly characterize Cryptosporidium and Giardia in rabbits in Heilongjiang Province, China. METHODOLOGY/PRINCIPAL FINDINGS: 378 fecal samples were obtained from rabbits in Heilongjiang Province. Cryptosporidium oocysts and Giardia cysts were detected using Sheather's sugar flotation technique and Lugol's iodine stain method, respectively. The infection rates of Cryptosporidium and Giardia were 2.38% (9/378 and 7.41% (28/378, respectively. Genotyping of Cryptosporidium spp. was done by DNA sequencing of the small subunit rRNA (SSU rRNA gene and all the nine isolates were identified as Cryptosporidium cuniculus. The nine isolates were further subtyped using the 60-kDa glycoprotein (gp60 gene and two subtypes were detected, including VbA32 (n = 3 and a new subtype VbA21 (n = 6. G. duodenalis genotypes and subtypes were identified by sequence analysis of the triosephosphate isomerase (TPI gene. The assemblage B (belonging to eight different subtypes B-I to B-VIII was found in 28 G. duodenalis-positive samples. CONCLUSIONS/SIGNIFICANCE: The rabbits have been infected with Cryptosporidium and Giardia in Heilongjiang Province. The results show that the rabbits pose a threat to human health in the studied areas. Genotypes and subgenotypes of C. cuniculus and G. duodenalis in this study might present the endemic genetic characterization of population structure of the two parasites.

  20. SPECTRUM OF OPPORTUNISTIC INFECTIONS IN HIV-AIDS PATIENTS

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    S S Madkar

    2012-10-01

    Full Text Available Aim: AIDS is characterized by a number of opportunistic infections which are responsible for high morbidity and mortality. The spectrum and distribution of opportunistic infections (OIs in AIDS patients is ever-expanding. This spectrum varies from continent to continent. The aim of the present study was to document the spectrum of OIs in HIV-infected patients in Ambajogai. Material and Method: 178 HIV positive symptomatic patients, either hospitalized or coming to ART (Antiretroviral Therapy centre in S.R.T.R. Medical College, Ambajogai, were included in the study for finding the spectrum of opportunistic infections. Result: The commonest opportunistic infection seen was tuberculosis (59% of patients, followed by oral candidiasis (37.6% of patients and parasitic diarrhea due to Cryptosporidium parvum(18 % of patients. It was observed that out of 178 patients, maximum 53.3% were in the age group of 29-38 years followed by 21% in the age group of 39-48 years. It was found that 77% were males and 23% were females, with male to female ratio is 3.3:1. Conclusion: This study demonstrates that tuberculosis is the commonest opportunistic infection seen in HIV patients. Clinicians should consider HIV in the differential diagnosis and management of all persons with tuberculosis.

  1. Cryptosporidium and Giardia in Africa: current and future challenges.

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  3. Prevalence and molecular identification of Cryptosporidium isolates from pet lizards and snakes in Italy

    Directory of Open Access Journals (Sweden)

    Rinaldi L.

    2012-11-01

    Full Text Available In order to acquire prevalence and genetic data on Cryptosporidium infections in captive lizards and snakes kept as pets, a survey was conducted on 150 individual reptiles from southern Italy. Fecal samples were preserved in 5% formalin and analyzed using a commercial immunofluorescence assay (IFA for the detection of Cryptosporidium oocysts and Giardia cysts. IFA revealed the presence of Cryptosporidium oocysts in nine of the 150 samples examined (6.0%, precisely in 6/125 snakes (4.8% and in 3/25 lizards (12.0%; all fecal samples tested negative for the presence of Giardia cysts. Molecular characterization based on nested PCR amplification and sequencing of the SSU-rRNA gene, revealed the presence of Cryptosporidium serpentis in three samples from snakes (Boa constrictor constrictor, Elapheguttata guttata guttata and Python molurus.

  4. Occurence of Cryptosporidium spp. in low quality water and on vegetables in Kumasi, Ghana

    DEFF Research Database (Denmark)

    Petersen, T. B.; Petersen, H. H.; Abaidoo, R. C.

    2014-01-01

    Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp. contaminat......Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp....... contamination level of river water, irrigation water and lettuce, 10L of water and 16 lettuce samples were collected four times in the period of, September – October 2013, with weekly intervals from six sample sites in and around Kumasi, Ghana. Oocysts were purified from water by sedimentation for 2 x 48 hours...... to characterize Cryptosporidium spp. positive samples were done by PCR amplification and sequencing of the SSU rRNA, the HSP70 and the GP60 genes after. Cryptosporidium oocysts were found in 75% of the water samples and on 43% of the lettuce with concentrations of 53 – 3268 per 10 L water and 11 – 118 oocyst per...

  5. Chemodiversity of Ladder-Frame Prymnesin Polyethers in Prymnesium parvum

    DEFF Research Database (Denmark)

    Rasmussen, Silas Anselm; Meier, Sebastian; Andersen, Nikolaj Gedsted;

    2016-01-01

    Blooms of the microalga Prymnesium parvum cause devastating fish kills worldwide, which are suspected to be caused by the supersized ladder-frame polyether toxins prymnesin-1 and -2. These toxins have, however, only been detected from P. parvum in rare cases since they were originally described two...

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

    Science.gov (United States)

    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.

  7. Prevalence and genetic characterizations of Cryptosporidium spp. in pre-weaned and post-weaned piglets in Heilongjiang Province, China.

    Directory of Open Access Journals (Sweden)

    Weizhe Zhang

    Full Text Available BACKGROUND: Cryptosporidium spp. are common intestinal protozoa of humans and animals. There have been few studies conducted on the molecular characterizations of pig-derived Cryptosporidium isolates worldwide, especially in China. Thus, the aim of the present study was to understand the prevalence, distribution and genotypes of Cryptosporidium in pigs in Heilongjiang Province, China. METHODOLOGY/PRINCIPAL FINDINGS: A total of 568 fecal samples from pre-weaned and post-weaned piglets were collected from eight pig farms from four areas of Heilongjiang Province. The average infection rate of Cryptosporidium was 1.6% (9/568 by microscopy. 113 samples were subjected to PCR amplification of the small subunit (SSU rRNA gene of Cryptosporidium, with 55.8% (63/113 being positive for Cryptosporidium. Cryptosporidium suis (n = 31 and C. scrofarumn (n = 32 were identified by DNA sequencing of the SSU rRNA gene. Three types of C. scrofarumn were found at the SSU rRNA locus, with one novel type being detected. Using species/genotype-specific primers for pig-adapted Cryptosporidium spp., 22 and 23 respectively belonged to C. suis and C. scrofarum mono-infections, with 18 co-infections detected. The infection peaks for C. suis (60%, 24/40 and C. scrofarum (51.2%, 21/41 were respectively found in the piglets of 5 to 8 weeks and more than 8 weeks. CONCLUSION/SIGNIFICANCE: The detection of C. suis and C. scrofarum in pre-weaned and post-weaned piglets has public health implications, due to the fact that the two species are both zoonotic Cryptosporidium. The novel C. scrofarum type detected may be endemic to China.

  8. Transport of Cryptosporidium parvum oocysts in soil columns following applications of raw and separated liquid slurry

    DEFF Research Database (Denmark)

    Petersen, Heidi Huus; Enemark, Heidi L.; Olsen, Annette;

    2012-01-01

    . Among leachate samples containing oocysts, 44/72 samples yielded viable oocysts as determined by a dye permeability assay (DAPI/PI) with the majority (41%) of viable oocysts found in leachate from soil columns with added liquid slurry. The number of viable oocysts was positively correlated (r=0...

  9. ESTIMATING MAXIMUM POSSIBLE ENVIRONMENTAL LOADING AMOUNTS OF CRYPTOSPORIDIUM PARVUM ATTRIBUTABLE TO BEEF CATTLE. (R828038)

    Science.gov (United States)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  10. Assessment of drugs against Cryptosporidium parvum using a simple in vitro screening method.

    Science.gov (United States)

    Armson, A; Meloni, B P; Reynoldson, J A; Thompson, R C

    1999-09-15

    A rapid semi-quantitative screening method was devised for assessing the anticryptosporidial and cytotoxic effects of putative chemotherapeutic compounds. The method is suitable as an initial rapid screening procedure from which compounds demonstrating anticryptosporidial activity can be identified for further analysis. It has the advantages of speed, low cost and concurrent assessment of anticryptosporidial and cytotoxic effects and allows accurate determination of minimum lethal concentrations. Of the 71 compounds screened, six completely inhibited cryptosporidial growth at 1 microM (monensin, salinomycin, alborixin, lasalocid, trifluralin and nicarbazin) and a further eight showed significant anticryptosporidial activity at 1 or 20 microM (halquinol, bleomycin, suramin, mitomycin, doxycycline hydrochloride, toltrazuril, chloroquine phosphate and teniposide). Twelve compounds were found to have some degree of cytotoxicity at 1 microM and a further 12 at 20 microM.

  11. PATHOGENESIS OF HUMAN AND BOVINE CRYPTOSPORIDIUM PARVUM IN GNOTOBIOTIC PIGS. (R826138)

    Science.gov (United States)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  12. PATHOGENESIS OF BOVINE (GENOTYPE 2) CRYPTOSPORIDIUM PARVUM STRAINS IN NEONATAL AND AGED GNOTOBIOTIC PIGS (R826138)

    Science.gov (United States)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  13. EFFECT OF LACTOBACILLUS AND BIFIDOBACTERIUM ON CRYPTOSPORIDIUM PARVUM OOCYST VIABILITY. (R826138)

    Science.gov (United States)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  14. Potential molecular tools for assessing the public health risk associated with waterborne Cryptosporidium oocysts.

    Science.gov (United States)

    Kothavade, Rajendra J

    2012-08-01

    The use of multiple barrier stages at water and wastewater treatment facilities allows for the effective removal of the vast majority of coliforms and other enteric and non-enteric microbes. Subsequent disinfection steps (chlorine, ozone and UV irradiation) are utilized to inactivate microbes that escape the preceding treatment stages. Most viruses, bacteria and protozoa, such as Giardia, are effectively inactivated by chlorination; however, Cryptosporidium is relatively more resistant to environmental conditions and to chlorination. Therefore, UV disinfection has been introduced at many water and wastewater treatment plants to increase log inactivation. Any accidental treatment failure may pose a significant risk to public health. Waterborne transmission of coccidian parasites such as Cryptosporidium and Giardia continues to be a major public health concern. No effective therapies currently exist to treat cryptosporidiosis and the global increase in immunocompromised populations has emphasized the need for water utilities and public health laboratories to have immediate and reliable access to highly sensitive test methods that can determine the host specificity, viability and infectivity of protozoa in the water supply. The most common method used for monitoring Cryptosporidium oocysts and Giardia cysts at intermediate treatment stages and in finished drinking water is the US EPA Method 1623. Although Cryptosporidium species are morphologically indistinguishable, they differ greatly in their host specificity and infectivity. Method 1623 provides quantitative information about Cryptosporidium and Giardia contamination but cannot distinguish between species for intervention purposes in outbreak situations, nor is this method reliable for determining whether the oocyst on the slide is infective for humans. Molecular methods have proven valuable in diagnosing infectious diseases, especially those for which the causative agent is difficult to grow in culture, and

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

    Science.gov (United States)

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

    2011-06-01

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

  16. Detection of Cryptosporidium oocysts in green mussels (Perna viridis) from shell-fish markets of Thailand.

    Science.gov (United States)

    Srisuphanunt, M; Wiwanitkit, Viroj; Saksirisampant, W; Karanis, P

    2009-09-01

    Mussels filter large volumes of water and can concentrate pathogenic organisms, which may act as potential vehicles of transmission to the consumer. A survey study was carried out to investigate the presence of Cryptosporidium protozoan parasites in green mussels (Perna viridis), the smussles pecies most destined for consumption in Thailand. In total, 56 samples were examined from Bangkok (n = 24) and Samut Prakan (n = 32) a wholesale shell-fish markets located at the mouth of the Chao Phraya River. The market for green mussels was closed to the mussel culture placed along the coastal line and this localization may have significant economical impact if the mussels' cultures are found contaminated. Cryptosporidium spp. oocysts were detected by the immunofluorescence antibody method (IFA) in 12.5% of the samples examined. The detection of Cryptosporidium oocysts in green mussels' population of Samut Prakan was higher (15.6%) than in Bangkok market (8.3%). These differences in positive samples from the two locations may be caused by physical, ecological and anthropogenic conditions. This could relay to different contamination levels of marine water by Cryptosporidium oocysts and consequently to contamination of harvested shellfish populations. The results demonstrate that the Cryptosporidium spp. oocysts were found indigenous in mussels from the coastal line of Thailand, indicating that mussels may act as a reservoir of Cryptosporidium foodborne infections for humans.

  17. Detection of Cryptosporidium oocysts in green mussels (Perna viridis from shell-fish markets of Thailand

    Directory of Open Access Journals (Sweden)

    Srisuphanunt M.

    2009-09-01

    Full Text Available Mussels filter large volumes of water and can concentrate pathogenic organisms, which may act as potential vehicles of transmission to the consumer. A survey study was carried out to investigate the presence of Cryptosporidium protozoan parasites in green mussels (Perna viridis, the smussles pecies most destined for consumption in Thailand. In total, 56 samples were examined from Bangkok (n = 24 and Samut Prakan (n = 32 a wholesale shell-fish markets located at the mouth of the Chao Phraya River. The market for green mussels was closed to the mussel culture placed along the coastal line and this localization may have significant economical impact if the mussels’ cultures are found contaminated. Cryptosporidium spp. oocysts were detected by the immunofluorescence antibody method (IFA in 12.5% of the samples examined. The detection of Cryptosporidium oocysts in green mussels’ population of Samut Prakan was higher (15.6% than in Bangkok market (8.3%. These differences in positive samples from the two locations may be caused by physical, ecological and anthropogenic conditions. This could relay to different contamination levels of marine water by Cryptosporidium oocysts and consequently to contamination of harvested shellfish populations. The results demonstrate that the Cryptosporidium spp. oocysts were found indigenous in mussels from the coastal line of Thailand, indicating that mussels may act as a reservoir of Cryptosporidium foodborne infections for humans.

  18. Het rendement van de detectiemethode voor Cryptosporidium en Giardia in water

    NARCIS (Netherlands)

    Schets FM; Medema GJ; Schijven JF; MGB

    2004-01-01

    Dutch drinking water legislation requires drinking water companies to perform a quantitative risk assessment for Cryptosporidium and Giardia. The risk of infection through consumption of drinking water should be below one infection in 10,000 persons per year. Risk assessment is based on the number

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

    Science.gov (United States)

    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.

  20. Chemodiversity of Ladder-Frame Prymnesin Polyethers in Prymnesium parvum

    DEFF Research Database (Denmark)

    Rasmussen, Silas Anselm; Meier, Sebastian; Andersen, Nikolaj Gedsted;

    2016-01-01

    Blooms of the microalga Prymnesium parvum cause devastating fish kills worldwide, which are suspected to be caused by the supersized ladder-frame polyether toxins prymnesin-1 and -2. These toxins have, however, only been detected from P. parvum in rare cases since they were originally described two......-HRMS) of 10 strains of P. parvum collected worldwide showed that only one strain produced the original prymnesin-1 and -2, whereas four strains produced the novel B-type prymnesin. In total 13 further prymnesin analogues differing in their core backbone and chlorination and glycosylation patterns could...

  1. Criptosporidiosis intestinal en niños con HIV/SIDA Intestinal cryptosporidiosis in HIV infected children

    Directory of Open Access Journals (Sweden)

    Graciela Barboni

    2008-06-01

    Full Text Available El Cryptosporidium parvum, protozoo parásito intracelular, infecta el epitelio gastrointestinal produciendo diarrea autolimitada en individuos inmunocompetentes pero potencialmente grave en pacientes inmunocomprometidos, especialmente en aquellos con Sida. En este trabajo se evaluó, durante un lap-so de 6 años, la incidencia de infección intestinal por C. parvum en una población pediátrica con HIV/Sida analizando las características clínicas e inmunológicas de la coinfección. Todos los pacientes iniciaron o continuaron el tratamiento antirretroviral de alta eficacia HAART durante el período de estudio, mientras que la infección intestinal fue tratada con azitromicina. La incidencia de criptosporidiosis fue de 13.7%. 33 de los 240 niños en seguimiento presentaron diarrea crónica de más de 14 días de evolución o recurrente, sin complicaciones hidroelectrolíticas. Los pacientes evaluados presentaron niveles porcentuales variables de células T CD4+ en sangre periférica, y la presencia del parásito no estuvo en relación con el compromiso inmunitario. Al momento del cuadro entérico 31 de los 33 pacientes tuvieron niveles plasmáticos de carga viral que superaban el límite de detección. Se observó eosinofilia leve o moderada en el 23% de los pacientes y la coinfección con otros parásitos fue detectada en 11 niños. No se obtuvieron diferencias significativas al relacionar el número de episodios intestinales con los estadios clínico-inmunológicos de los pacientes. La correcta implementación del HAART con la subsecuente restauración de la función inmune se relacionaría con la ausencia de cuadros diarreicos agudos y de las complicaciones hidroelectrolíticas derivadas de la coinfección con C. parvum.Cryptosporydium parvum is an intracellular parasite that infects gastrointestinal epithelium and produces diarrhea that is self-limited in immunocompetent persons but potentially life-threatening in immunocompromised

  2. [Cryptosporidium spp. and Giardia spp.--environmental studies in Poland].

    Science.gov (United States)

    Bajer, Anna; Bednarska, Małgorzata; Siński, Edward

    2009-01-01

    Cryptosporidium spp. and Giardia spp. are intestinal protozoan parasites of humans and many other species of mammals. The aim of this article was to summarize the last twenty years of research on the environmental distribution of these parasites, with a particular emphasis on the natural reservoir of invasion and human infections in Poland. The prevalence of Cryptosporidium and Giardia has been studied in different groups of humans, in wildlife, pets and farm animals and in environmental samples. Current knowledge on the distribution of zoonotic and non-zoonotic species/genotypes in reservoir hosts and environmental samples has been summarized. The usefulness of different methods for the detection and identification of the parasites in different types of samples has been presented. Due to the wide distribution and high prevalence of both species in a range of hosts and possible vectors involved in mechanical transmission, the overall risk of outbreaks of cryptosporidiosis and giardiosis in Poland has been assessed as relatively high.

  3. Transport of Cryptosporidium oocysts in porous media: Role of straining and physicochemical filtration

    Science.gov (United States)

    Tufenkji, N.; Miller, G.F.; Ryan, J.N.; Harvey, R.W.; Elimelech, M.

    2004-01-01

    The transport and filtration behavior of Cryptosporidium parvum oocysts in columns packed with quartz sand was systematically examined under repulsive electrostatic conditions. An increase in solution ionic strength resulted in greater oocyst deposition rates despite theoretical predictions of a significant electrostatic energy barrier to deposition. Relatively high deposition rates obtained with both oocysts and polystyrene latex particles of comparable size at low ionic strength (1 mM) suggest that a physical mechanism may play a key role in oocyst removal. Supporting experiments conducted with latex particles of varying sizes, under very low ionic strength conditions where physicochemical filtration is negligible, clearly indicated that physical straining is an important capture mechanism. The results of this study indicate that irregularity of sand grain shape (verified by SEM imaging) contributes considerably to the straining potential of the porous medium. Hence, both straining and physicochemical filtration are expected to control the removal of C. parvum oocysts in settings typical of riverbank filtration, soil infiltration, and slow sand filtration. Because classic colloid filtration theory does not account for removal by straining, these observations have important implications with respect to predictions of oocyst transport.

  4. Chitosan and metal salt coagulant impacts on Cryptosporidium and microsphere removal by filtration.

    Science.gov (United States)

    Brown, Trevor J; Emelko, Monica B

    2009-02-01

    Maintenance of appropriate chemical pretreatment is a critical component of ensuring proper filtration performance. Pilot-scale in-line filtration studies were performed to investigate the relative impacts of chitosan, alum, and FeCl(3) coagulation on the removal of Cryptosporidium parvum oocysts and oocyst-sized polystyrene microspheres by granular media filtration. Similar removals of oocysts and microspheres were achieved when optimal coagulant doses were utilized. Sub-optimal alum and FeCl(3) coagulation resulted in a deterioration filter effluent turbidity (0.2-0.3NTU) and total particle counts (30-100 total particles > or =2microm/mL) that were accompanied by reduced (by approximately 2-3-log) median oocyst and microsphere removals by filtration. At all doses investigated, chitosan coagulation resulted in excellent turbidity and particle reductions by filtration. Nonetheless, chitosan coagulation at doses of 0.1, 0.5, and 1.0mg/L did not result in appreciable improvements in C. parvum oocyst removal relative to complete coagulation failure (median oocyst removals were alum and FeCl(3) coagulation, but not chitosan coagulation.

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

    Science.gov (United States)

    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.

  6. Cryptosporidium en Giardia in Nederlandse zwembaden

    NARCIS (Netherlands)

    Schets FM; Engels GB; Leenen EJTM; MGB

    2003-01-01

    Swimming-pool associated outbreaks of cryptosporidiosis have been frequently reported in the UK and USA. Cryptosporidium oocysts could sometimes be detected in the pool water or the filter backwash water in cases where the source of the outbreak was confirmed. The occurrence of Cryptosporidium and G

  7. Emissie van Cryptosporidium en Giardia door landbouwhuisdieren

    NARCIS (Netherlands)

    Schrijven JF; Bruin HAM de; Engels GB; Leenen EJTM; MGB

    1999-01-01

    In this study, the relative contributions of the pathogenic protozoa Cryptosporidium and Giardia by manure of farm animals in The Netherlands to the total yearly environmental load was studied. Manure of veal calves forms a very large source of Cryptosporidium (1.5 m 10 square 16 oocysts per year) a

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

    Directory of Open Access Journals (Sweden)

    Joyce Siwila

    2012-02-01

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

  9. 猪源隐孢子虫卵囊的分离及基因型鉴定%Isolation and genotype identification of Cryptosporidium oocysts in pigs

    Institute of Scientific and Technical Information of China (English)

    陈甫; 黄克和

    2011-01-01

    To investigate the genotypes of Cryptosporidium parasiting in pigs in Nanjing,Shanghai and Hefei, the total RNA of the oocysts from 6 positive feces was extracted using the TRIZOL reagent,and the partial genes of the small subunit ribosomal RNA (SSU rRNA) were amplified by the reverse transcription polymerase chain reaction (RT-PCR). SSU rRNA gene fragments of the isolates were deposited in the GenBank database under accession NO. DQ855266,DQ855267 and compared with sequences of various Cryptosporidium spp. and genotypes on the NCBI server using the BLAST and DNAStar software by the analysis of the homologous rate and phylogenetic tree. The results showed that the SSU rRNA gene fragments,compared with sequences of various Cryptosporidium spp. and genotypes,belonged to the species C. parvum with 94 %-100 % homologous rate, and were most similar to the C. parvum “mouse” genotype with 99. 8%-100 % homologous rate. The phylogenetic tree and sequence distances gen erated showed that those isolates belong to the C. parvum groups,and lie in the same divarication with C. parvum “mouse” genotype. It is concluded that those isolates are clearly distinct from other sequences as to merit species status, which is C. parvum “mouse” genotype from pigs. There might be a cross-transmission between pigs and mice.%应用RT-PCR方法对南京、上海和合肥猪源隐孢子虫卵囊SSU rRNA部分序列进行扩增,产物测序后提交GenBank,收录号为DQ855266、DQ855267;用BLAST和DNAStar软件与GenBank参考序列进行比较,分析其同源性,绘制系统发育进化树,结合卵囊形态学观察和对小鼠、大鼠、兔、山羊和鸡的传染性试验确定隐孢子虫种类或基因型.结果表明,3地区猪源隐孢子虫分离株与微小隐孢子虫(C.parvum)同源性达94%~100%,与C.parvum"mouse"型有99.8%~100%的同源性,并处于进化树的同一分支.因此,3地区猪源隐孢子虫是C.parvum"mouse"型,提示猪和鼠之间存在交叉传播的可能.

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

    Science.gov (United States)

    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.

  11. Genotyping of Cryptosporidium spp. from free-living wild birds from Brazil.

    Science.gov (United States)

    Sevá, Anaiá da Paixão; Funada, Mikaela Renata; Richtzenhain, Leonardo; Guimarães, Marta Brito; Souza, Sheila de Oliveira; Allegretti, Luciana; Sinhorini, Juliana Anaya; Duarte, Vanessa Vertematti; Soares, Rodrigo Martins

    2011-01-10

    In wild and domestic birds, cryptosporidiosis is often associated with infections by Cryptosporidium galli, Cryptosporidium baileyi and Cryptosporidium meleagridis. In addition to these species, a number of avian Cryptosporidium species yet to be fully characterized are commonly found among exotic and wild avian isolates. The present study aimed to detect and identify samples of Cryptosporidium spp. from free-living wild birds, in order to contribute to the knowledge of the variability of this parasite in the free-living population of Brazil. Stool samples were collected from 242 birds, with the following proportions of individuals: 50 Emberizidae (20.7%), 112 Psittacidae (46.3%), 44 Cardinalidae (18.2%), 12 Turdidae (5.0%), eight Ramphastidae (3.3%), seven Icteridae (2.9%), three Estrilididae (1.2%), two Contigidae (0.8%), two Thraupidae (0.8%) and two Fringilidae (0.8%). Among the 242 fecal samples from wild birds, 16 (6.6%) were positive for the presence of oocysts of Cryptosporidium. Molecular characterization of the 16 samples of Cryptosporidium, were performed with phylogenetic reconstructions employing 292 positions of 18S rDNA. None of the samples of birds was characterized as C. meleagridis. C. galli was identified in one rufous-bellied thrush (Turdus rufiventris), five green-winged saltators (Saltator similis), one slate-coloured seedeater (Sporophila schistacea), one goldfinch (Carduelis carduelis) and three saffron finches (Sicalis flaveola). One goldfinch isolate, one buffy-fronted seedeater (Sporophila frontalis), one red-cowled cardinal (Paroaria dominicana) and one other saffron finch (S. flaveola) were identified as C. baileyi. Avian genotype II was found in an isolate from a white-eyed parakeet (Aratinga leucophthalma). Clinical symptoms of cryptosporidiosis in birds have already been described and the number of wild birds which were shedding parasites was high. Therefore, further epidemiological research and disease surveillance of birds in the

  12. Occurrence of Cryptosporidium spp. oocysts in low quality water and on vegetables irrigated with low quality water in Kumasi, Ghana

    DEFF Research Database (Denmark)

    Petersen, Tobias Boel; Petersen, Heidi Huus; Robert C., Abaidoo

    Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp....... contamination level of river water, irrigation water and lettuce, 10L of water and 16 lettuce samples were collected four times in the period September – October 2013, with weekly intervals from six sample sites in and around Kumasi, Ghana. Oocysts were purified from water by sedimentation for 2 x 48 hours...... off the slides and attempts to characterize Cryptosporidium spp. positive samples were done by PCR amplification and sequencing of the SSU rRNA, the HSP70 and the GP60 genes. Cryptosporidium oocysts were found in 75% of the water samples and on 43% of the lettuce with concentrations of 53 – 3,268 per...

  13. Occurrence of Cryptosporidium spp. oocysts in low quality water and on vegetables irrigated with low quality water in Kumasi, Ghana

    DEFF Research Database (Denmark)

    Petersen, Tobias B; Petersen, Heidi H.; Abaidoo, Robert C.

    Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp. contaminat......Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp....... contamination level of river water, irrigation water and lettuce, 10L of water and 16 lettuce samples were collected four times in the period September – October 2013, with weekly intervals from six sample sites in and around Kumasi, Ghana. Oocysts were purified from water by sedimentation for 2 x 48 hours...... off the slides and attempts to characterize Cryptosporidium spp. positive samples were done by PCR amplification and sequencing of the SSU rRNA, the HSP70 and the GP60 genes. Cryptosporidium oocysts were found in 75% of the water samples and on 43% of the lettuce with concentrations of 53 – 3,268 per...

  14. The public health and clinical significance of Giardia and Cryptosporidium in domestic animals.

    Science.gov (United States)

    Thompson, R C Andrew; Palmer, Carlysle S; O'Handley, Ryan

    2008-07-01

    Giardia and Cryptosporidium are common enteric parasites of domestic animals, particularly dogs, cats and livestock. Their occurrence is of potential significance from both clinical and public health perspectives yet, until recently, confusion over the taxonomy of these organisms prevented a clear understanding of the epidemiology of infections with both Giardia and Cryptosporidium. The recent application of molecular epidemiological tools has helped to resolve taxonomic issues, allowing cycles of transmission to be determined. In addition, advances have been made in elucidating mechanisms associated with pathogenesis, whereas only limited progress has been achieved in the areas of chemotherapy and prophylaxis.

  15. Possible Transmission of Cryptosporidium canis among Children and a Dog in a Household▿

    OpenAIRE

    Xiao, Lihua; Cama, Vitaliano A.; Cabrera, Lilia; Ortega, Ynes; Pearson, Julie; Gilman, Robert H.

    2007-01-01

    In a longitudinal cohort diarrhea study, a girl living in Lima, Peru, and her brother and dog were diagnosed with Cryptosporidium canis infections during the same period. Both children had transient diarrhea, but the dog was asymptomatic. This is the first report of possible transmission of cryptosporidiosis between humans and dogs.

  16. Acid-fast lipids are important structural components of oocyst walls of Cryptosporidium, Toxoplasma, and Eimeria

    Science.gov (United States)

    Coccidia are protozoan parasites that cause significant human disease and are of major agricultural importance. Cryptosporidium spp.cause diarrhea in humans and animals, while congenital Toxoplasma infections causes blindness and death. Eimeria kills chickens, so all poultry feed contain antibioti...

  17. [Semi-aquatic animals as a source of water contamination with Cryptosporidium and Giardia].

    Science.gov (United States)

    Bajer, Anna; Bednarska, Małgorzata; Paziewska, Anna; Romanowski, Jerzy; Siński, Edward

    2008-01-01

    Cryptosporidium and Giardia spp. are parasitic protozoa localized in the alimentary tract of many animal species and humans. Each of these parasite species produces very resistant invasive forms (cysts and oocysts) excreted to the environment with feces of infected hosts. Water contaminated with cysts/oocysts constitutes one of the main transmission routes and is responsible for the majority of infections in humans. Cryptosporidium and Giardia spp. were found in many different species of animals, including livestock, pets and free living animals. The aim of our study was to determine the prevalence of these protozoa in selected species of semi-aquatic mammals and to estimate their role in water contamination. In years 1996-98 the prevalence of Cryptosporidium and Giardia infections was high in muskrats (Ondatra zibethicus) (58 and 87%, respectively). The origin of animals (farmed or free living) affected the prevalence of both parasites in European beavers (Castor fiber). The prevalence of infection increased in second period of study and was 4 and 19% for Cryptosporidium and 0 and 8% for Giardia spp. in the two studied periods, respectively. Both parasite species were also identified in water vole (Arvicola terrestris) and rat (Rattus norvegicus).

  18. Occurrence of Cryptosporidium and Giardia in domestic animals in peri-urban communities of Kafue district, Zambia

    DEFF Research Database (Denmark)

    Siwila, J.; Phiri, I. G. K.; Enemark, Heidi L.

    2013-01-01

    Cryptosporidium spp. and Giardia duodenalis are important parasites infecting a wide range of domestic animals worldwide. The aim of the present study was to determine the occurrence of Cryptosporidium spp. and Giardia parasites in different domestic animals living in close contact with humans...... within rural/semiurban communities in Kafue district in Zambia. A single faecal sample per animal was collected from pigs, goats, dogs, ducks, chickens and pigeons and analysed by Merifluor Cryptosporidium/Giardia immunofluorescence antibody assay for the simultaneous detection of these parasites....../148), goats (5.9%; 1/17), dogs (25.0%; 5/20) and ducks (6.7%; 2/30). Diarrhoea was not associated with either infection. Age was also not associated with either infection except in dogs where Giardia infection was only detected in animals aged less than six months (p=0.009). It is concluded from this study...

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

    Science.gov (United States)

    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.

  20. Waterborne transmission of Giardia and Cryptosporidium at river beaches in Southern Europe (Portugal).

    Science.gov (United States)

    Júlio, Cláudia; Sá, Cátia; Ferreira, Idalina; Martins, Susana; Oleastro, Mónica; Angelo, Helena; Guerreiro, José; Tenreiro, Rogério

    2012-09-01

    Giardia and Cryptosporidium are the most frequent enteric protozoa causing gastroenteritis in humans worldwide. Intense recreational activity at Portuguese river beaches triggered the opportunity for a 2-year seasonal survey of 19 large river basin beaches. A total of 74 samples were collected and processed according to USEPA Method 1623 to detect Cryptosporidium and Giardia (oo)cysts. Faecal indicators (thermotolerant/total coliforms, Escherichia coli, and enterococci) and physicochemical parameters were also analysed according to the EU Bath Water Directive (BWD). Results pointed to a widespread presence of these protozoa at Portuguese river beaches. The percentage of samples testing positive for Giardia and Cryptosporidium were 85 and 82% respectively, with no significant differences between wet and dry seasons (p > 0.05). Although Portuguese river beaches present a very low exposure risk for infection with Giardia and Cryptosporidium (under 10(-3)), a few particular cases revealed values over 0.2%, and were related to stormy wet events. The correlation between levels of Giardia and thermotolerant coliforms, E. coli and enterococci, was high (r ≥ 0.87, p Giardia and Cryptosporidium whenever the values of those faecal indicators approach the maximum allowed level of the EU BWD.

  1. Occurrence of Cryptosporidium spp. and Giardia duodenalis in red foxes (Vulpes vulpes in Bosnia and Herzegovina

    Directory of Open Access Journals (Sweden)

    Adnan Hodžić

    2014-10-01

    Full Text Available The aim of this study was to determine the occurrence and distribution of Cryptosporidium spp. and Giardia duodenalis among free-living red foxes (Vulpes vulpes in Bosnia and Herzegovina. For this purpose, a total of 123 fecal samples fromred foxes, shot during hunting seasons between January 2011 and March 2012 were examined using immunofluorescent microscopy. Overall, observed prevalences of Cryptosporidium spp. and Giardia duodenalis were 3.2 % (4/123 and 7.3% (9/123, respectively. The results show that foxes might play the role of potential reservoirs of Cryptosporidium and Giardia parasites, but further molecular analysis are necessary to elucidate the source of infection, routes of transmission and zoonotic potential of these two pathogens.

  2. Common occurrence of zoonotic pathogen Cryptosporidium meleagridis in broiler chickens and turkeys in Algeria.

    Science.gov (United States)

    Baroudi, Djamel; Khelef, Djamel; Goucem, Rachid; Adjou, Karim T; Adamu, Haileeyesus; Zhang, Hongwei; Xiao, Lihua

    2013-09-23

    Only a small number of birds have been identified by molecular techniques as having Cryptosporidium meleagridis, the third most important species for human cryptosporidiosis. In this study, using PCR-RFLP analysis of the small subunit (SSU) rRNA gene, we examined the ileum of 90 dead chickens from 23 farms and 57 dead turkeys from 16 farms in Algeria for Cryptosporidium spp. C. meleagridis-positive specimens were subtyped by sequence analysis of the 60 kDa glycoprotein gene. Cryptosporidium infection rates were 34% and 44% in chickens and turkeys, respectively, with all positive turkeys (25) and most positive chickens (26/31) having C. meleagridis. All C. meleagridis specimens belonged to a new subtype family. The frequent occurrence of C. meleagridis in chickens and turkeys illustrates the potential for zoonotic transmission of cryptosporidiosis in Algeria.

  3. Prymnesium parvum exotoxins affect the grazing and viability of the calanoid copepod Eurytemora affinis

    DEFF Research Database (Denmark)

    Sopanen, S.; Koski, Marja; Uronen, P.

    2008-01-01

    The calanoid copepod Eurytemora affinis from the northern Baltic Sea was exposed to cell-free filtrates of the toxic haptophyte Prymnesium parvum as well as to cell mixtures of P. parvum and Rhodomonas salina. To test the effects of P. parvum exudates and allelopathy on selective grazers, copepods...

  4. Detection of Cryptosporidium - specific coproantigen in human immunodeficiency virus/acquired immunodeficiency syndrome patients by using a commercially available immunoenzymatic assay

    Directory of Open Access Journals (Sweden)

    Claudio Vieira Silva

    2003-12-01

    Full Text Available The aim of this study was to verify the occurrence of Cryptosporidium infection in 52 human immunodeficiency virus (HIV/acquired immunodeficiency syndrome (AIDS patients (group 1 and 38 clinically healthy individuals (group 2 by using enzyme immunoassay (EIA. All fecal samples collected were submitted to the Baermann, Lutz, and Ritchie methods, the Safranin/Methylene Blue, and Weber's chromotrope modified Trichrome staining techniques, and EIA. In group 1, parasitological staining techniques and EIA were both positive for Cryptosporidium sp. infection in 3/52 (5.8% samples and both negative in 45/52 (86.5% samples, while 4/52 (7.7% samples were positive in EIA and negative in parasitological staining techniques. Concerning group 2, all samples were negative by EIA and microscopy for Cryptosporidium infection. In conclusion, EIA may be an alternative method for detecting Cryptosporidium-specific coproantigen in HIV/AIDS patients.

  5. Cryptosporidium in pet snakes from Italy: molecular characterization and zoonotic implications.

    Science.gov (United States)

    Díaz, P; Rota, S; Marchesi, B; López, C; Panadero, R; Fernández, G; Díez-Baños, P; Morrondo, P; Poglayen, G

    2013-10-18

    To provide information on the occurrence of Cryptosporidium species and genotypes in captive snakes from Italy, faecal specimens from 120 snakes belonging to 13 different genera of the families Boidae, Colubridae and Pythonidae were collected. Faecal samples were taken from the ground of the terrarium when available; otherwise cloacal cotton swabs were used. No clinical signs of cryptosporidiosis were observed in any animal at the time of sampling. Samples were examined for the presence of Cryptosporidium by using a direct immunofluorescence antibody test (IFAT) and two-step nested PCR at the small subunit (SSU) rRNA locus. PCR-positive samples were genotyped by restriction fragment length polymorphism (RFLP) analysis with the endonucleases SspI and VspI. By IFAT, 42 out of 120 snakes (35.0%) were found to be shedding Cryptosporidium oocysts. A significant higher percentage of positive ophidians were detected by using faecal specimens obtained from the terrarium (55.5%) than by cloacal cotton swabs (29.0%). SSU rRNA gene products were obtained from 25 isolates. Twenty samples tested positive to both microscopy and molecular techniques. Our data reveal a wide extent of cryptosporidial infections in snake-food animals since most of the identified isolates belonged to Cryptosporidium species, some of them with zoonotic potential, considered specific for rodents and resulting from ingestion of infected preys. The reptilian-specific species Cryptosporidium serpentis was identified in only one isolate. The common presence of reptile non-specific and, in some cases, zoonotic Cryptosporidium oocysts in snake faeces should to be taken into consideration in order to avoid the misidentification of the protozoan as well as the possible public health implications.

  6. Determination of the recovery efficiency of cryptosporidium oocysts and giardia cysts from seeded bivalve mollusks.

    Science.gov (United States)

    Schets, Franciska M; van den Berg, Harold H J L; de Roda Husman, Ana Maria

    2013-01-01

    The intestinal parasites Cryptosporidium and Giardia are transmitted by water and food and cause human gastroenteritis. Filter-feeding bivalve mollusks, such as oysters and mussels, filter large volumes of water and thus concentrate such pathogens, which makes these bivalves potential vectors of disease. To assess the risk of infection from consumption of contaminated bivalves, parasite numbers and parasite recovery data are required. A modified immunomagnetic separation (IMS) procedure was used to determine Cryptosporidium oocyst and Giardia cyst numbers in individually homogenized oysters (Crassostrea gigas) and mussels (Mytilus edulis). About 12% of the commercial bivalves were positive, with low (oo)cyst numbers per specimen. The recovery efficiency of the IMS procedure was systematically evaluated. Experiments included seeding of homogenized bivalves and whole animals with 100 to 1,000 (oo)cysts. Both seeding procedures yielded highly variable recovery rates. Median Cryptosporidium recoveries were 7.9 to 21% in oysters and 62% in mussels. Median Giardia recoveries were 10 to 25% in oysters and 110% in mussels. Giardia recovery was significantly higher than Cryptosporidium recovery. (Oo)cysts were less efficiently recovered from seeded whole animals than from seeded homogenates, with median Cryptosporidium recoveries of 5.3% in oysters and 45% in mussels and median Giardia recoveries of 4.0% in oysters and 82% in mussels. Both bivalve homogenate seeding and whole animal seeding yielded higher (oo)cyst recovery in mussels than in oysters, likely because of the presence of less shellfish tissue in IMS when analyzing the smaller mussels compared with the larger oysters, resulting in more efficient (oo)cyst extraction. The data generated in this study may be used in the quantitative assessment of the risk of infection with Cryptosporidium or Giardia associated with the consumption of raw bivalve moll