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Sample records for additional cryptosporidium treatment

  1. 40 CFR 141.711 - Filtered system additional Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Treatment for Cryptosporidium Treatment Technique Requirements § 141.711 Filtered system additional Cryptosporidium treatment requirements. (a) Filtered systems must provide the level of additional treatment for....5-log. (b)(1) Filtered systems must use one or more of the treatment and management options...

  2. 40 CFR 141.711 - Filtered system additional Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 23 2014-07-01 2014-07-01 false Filtered system additional Cryptosporidium treatment requirements. 141.711 Section 141.711 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium...

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

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Cryptosporidium treatment requirements. 141.711 Section 141.711 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Treatment Technique Requirements § 141.711 Filtered system...

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

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Cryptosporidium treatment requirements. 141.711 Section 141.711 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Treatment Technique Requirements § 141.711 Filtered system...

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

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Cryptosporidium treatment requirements. 141.711 Section 141.711 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Treatment Technique Requirements § 141.711 Filtered system...

  6. Additional novel Cryptosporidium genotypes in ornamental fishes.

    PubMed

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

    2012-12-21

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

  7. Cryptosporidium

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  8. 40 CFR 141.712 - Unfiltered system Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 23 2011-07-01 2011-07-01 false Unfiltered system Cryptosporidium treatment requirements. 141.712 Section 141.712 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Treatment...

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

    PubMed

    Nasser, Abidelfatah M

    2016-02-01

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

  10. 40 CFR 141.712 - Unfiltered system Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... violation of the treatment technique requirement. (2) Systems that use UV light and fail to achieve the..., ozone, or UV as described in § 141.720 to meet the Cryptosporidium inactivation requirements of...

  11. 40 CFR 141.712 - Unfiltered system Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... violation of the treatment technique requirement. (2) Systems that use UV light and fail to achieve the..., ozone, or UV as described in § 141.720 to meet the Cryptosporidium inactivation requirements of...

  12. 40 CFR 141.712 - Unfiltered system Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... violation of the treatment technique requirement. (2) Systems that use UV light and fail to achieve the..., ozone, or UV as described in § 141.720 to meet the Cryptosporidium inactivation requirements of...

  13. 40 CFR 141.713 - Schedule for compliance with Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Cryptosporidium treatment requirements. 141.713 Section 141.713 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Treatment Technique Requirements § 141.713 Schedule for compliance...

  14. 40 CFR 141.713 - Schedule for compliance with Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Cryptosporidium treatment requirements. 141.713 Section 141.713 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Treatment Technique Requirements § 141.713 Schedule for compliance...

  15. 40 CFR 141.713 - Schedule for compliance with Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Cryptosporidium treatment requirements. 141.713 Section 141.713 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Treatment Technique Requirements § 141.713 Schedule for compliance...

  16. 40 CFR 141.713 - Schedule for compliance with Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Cryptosporidium treatment requirements. 141.713 Section 141.713 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium Treatment Technique Requirements § 141.713 Schedule for compliance...

  17. CONTROL OF CRYPTOSPORIDIUM OOCYSTS BY STEADY-STATE CONVENTIONAL TREATMENT

    EPA Science Inventory

    Pilot-scale experiments have been performed to assess the ability of conventional treatment to control Cryptosporidium oocysts under steady-state conditions. The work was performed with a pilot plant that was designed to minimize flow rates and, as a result, the number of oocyst...

  18. Effect of water treatment processes on Cryptosporidium infectivity.

    PubMed

    Keegan, Alexandra; Daminato, David; Saint, Christopher P; Monis, Paul T

    2008-03-01

    Conventional water treatment processes have the ability to remove Cryptosporidium oocysts through coagulation, flocculation, sedimentation and filtration, provided there is efficient management of plant performance. The potential exists for the breakthrough of oocysts through the treatment train. The effect of the water treatment chemical aluminium sulphate (alum) on Cryptosporidium oocyst infectivity has been assessed using an assay that combines cell culture and real-time polymerase chain reaction techniques. The infectivity of fresh and temperature-aged oocysts (stored up to 6 months at 4 or 15 degrees C) was unaffected by exposure to a range of doses of alum in standard jar test procedures and dissolved air flotation processes and subsequent exposure to chlorine or chloramine. Removal efficiencies and infectivity measures are important in determining risk to public health and will reflect the ability of water treatment plants to act as a barrier to these pathogens. PMID:18067945

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

    PubMed

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

    2013-01-01

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

  20. 40 CFR 141.713 - Schedule for compliance with Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 23 2014-07-01 2014-07-01 false Schedule for compliance with Cryptosporidium treatment requirements. 141.713 Section 141.713 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for Cryptosporidium...

  1. Occurrence of Cryptosporidium in a wastewater treatment plant in North Germany.

    PubMed

    Ajonina, Caroline; Buzie, Christopher; Ajonina, Irene U; Basner, Alexander; Reinhardt, Heiko; Gulyas, Holger; Liebau, Eva; Otterpohl, Ralf

    2012-01-01

    Cryptosporidium parvum is one of the most common human parasitic protozoa and is responsible for many waterborne outbreaks in several industrialized countries. The oocyst, which is the infective form, is known to be highly resistant to wastewater treatment procedures and represents a potential hazard to human populations through contaminated raw or treated wastewater. In this investigation, the occurrence of Cryptosporidium in wastewater samples was monitored and removal efficiency was assessed. Treated (effluent) and untreated (influent) wastewater samples were collected seasonally over a period of 2 years. Oocysts were repeatedly detected in influent and effluent samples collected from the treatment plant during all sampling seasons, with a mean concentration of 782 oocysts/L. The seasonal distribution showed that oocysts are predominant during autumn and winter. Molecular analyses via the small (18S) subunit of rRNA amplification and subsequent sequencing with an objective of characterizing the oocysts revealed that Cryptosporidium parvum was the dominant Cryptosporidium parasite present in wastewater. PMID:23095153

  2. Interaction between Cryptosporidium oocysts and water treatment coagulants.

    PubMed

    Bustamante, H A; Shanker, S R; Pashley, R M; Karaman, M E

    2001-09-01

    The electrokinetic properties of gamma-irradiated Cryptosporidium oocysts in the presence of coagulants (ferric chloride and alum) and coagulant aids (DADMAC based cationic polyelectrolytes) have been studied. The zeta potential of the oocysts was unaffected by the addition of ferric chloride at all pH values (3-10) studied. Addition of alum resulted in reversal of the oocysts charge, which suggests that the initial stage in the coagulation process leading to floc formation proceeds via the adsorption of hydrolysed aluminium species. The cationic polyelectrolyte Magnafloc LT35 was adsorbed onto iron flocs at doses of 0.1 mg/L even against an electrostatic barrier. The cationic polyelectrolyte only adsorbed and caused charge reversal at the oocyst surface at around 0.4 mg/L, suggesting a lower affinity for this surface. These results indicate that the oocysts, unlike inorganic colloidal materials such as metal oxides, appear to possess a lower surface density of active or charged sites. The lower density of sites, combined with the rapid precipitation of iron salts, may be responsible for the lack of specific adsorption of either hydroxylated ferric species or primary iron hydroxide particles on the oocysts. Further, this suggests that a process of sweep flocculation, where oocysts are engulfed in flocs during coagulation and floc formation, is the more likely mechanism involved. By comparison, it is likely that the specific interaction of hydrolysed aluminium species with the oocysts surface would result in a stronger link at the oocyst-floc interface and that the flocculation process may initially proceed via charge neutralisation. PMID:11487115

  3. REMOVAL OF CRYPTOSPORIDIUM AND GIARDIA THROUGH CONVENTIONAL WATER TREATMENT AND DIRECT FILTRATION

    EPA Science Inventory

    Pilot- and full-scale evaluations of Giardia and Cryptosporidium cyst removal through direction filtration and conventional water treatment were conducted by the Utah Department of Environmental Quality. Cysts were seeded continuously in a step dose at a 0.5 gpm pilot plant and i...

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

    PubMed

    Petterson, S; Roser, D; Deere, D

    2015-09-01

    It is proposed that the next revision of the Australian Drinking Water Guidelines will include 'health-based targets', where the required level of potable water treatment quantitatively relates to the magnitude of source water pathogen concentrations. To quantify likely Cryptosporidium concentrations in southern Australian surface source waters, the databases for 25 metropolitan water supplies with good historical records, representing a range of catchment sizes, land use and climatic regions were mined. The distributions and uncertainty intervals for Cryptosporidium concentrations were characterized for each site. Then, treatment targets were quantified applying the framework recommended in the World Health Organization Guidelines for Drinking-Water Quality 2011. Based on total oocyst concentrations, and not factoring in genotype or physiological state information as it relates to infectivity for humans, the best estimates of the required level of treatment, expressed as log10 reduction values, ranged among the study sites from 1.4 to 6.1 log10. Challenges associated with relying on historical monitoring data for defining drinking water treatment requirements were identified. In addition, the importance of quantitative microbial risk assessment input assumptions on the quantified treatment targets was investigated, highlighting the need for selection of locally appropriate values. PMID:26322774

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

    SciTech Connect

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

    1996-11-01

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

  6. Occurrence of Cryptosporidium, Giardia, and Cyclospora in influent and effluent water at wastewater treatment plants in Arizona.

    PubMed

    Kitajima, Masaaki; Haramoto, Eiji; Iker, Brandon C; Gerba, Charles P

    2014-06-15

    We investigated the occurrence of Cryptosporidium, Giardia, and Cyclospora at two wastewater treatment plants (WWTPs) in Arizona over a 12-month period, from August 2011 to July 2012. Influent and effluent wastewater samples were collected monthly, and protozoan (oo)cysts were concentrated using an electronegative filter, followed by the detection of protozoa using fluorescent microscopy (Cryptosporidium oocysts and Giardia cysts) and PCR-based methods (Cryptosporidium spp., Giardia intestinalis, and Cyclospora cayetanensis). The concentration of Giardia cysts in the influent was always higher than that of Cryptosporidium oocysts (mean concentration of 4.8-6.4×10(3) versus 7.4×10(1)-1.0×10(2)(oo)cysts/l) with no clear seasonality, and log10 reduction of Giardia cysts was significantly higher than that of Cryptosporidium oocysts for both WWTPs (P<0.05). Log10 reduction of Giardia cysts at the WWTP utilizing activated sludge was significantly higher than the other WWTP using trickling filter (P=0.014), while no statistically significant difference between the two WWTPs was observed for the log10 reduction of Cryptosporidium oocysts (P=0.207). Phylogenetic analysis revealed that G. intestinalis strains identified in wastewater belonged to two assemblages, AII and B, which are potentially infectious to humans. C. cayetanensis was also detected from both influent and effluent using a newly developed quantitative PCR, with the highest influent concentration of 1.2×10(4)copies/l. Our results demonstrated that these protozoan pathogens are prevalent in the study area and that efficacy of the conventional wastewater treatment processes at physically removing (oo)cysts is limited. PMID:24695096

  7. 40 CFR 141.712 - Unfiltered system Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for... water monitoring required under § 141.701(a), unfiltered systems must calculate the arithmetic mean of... initial source water monitoring based on the schedule in § 141.701(c). (2) Following completion of...

  8. Antibiotics for the treatment of Cholera, Shigella and Cryptosporidium in children

    PubMed Central

    2013-01-01

    Background Diarrhea is a major contributor to the burden of morbidity and mortality in children; it accounts for a median of 11% of all deaths among children aged less than 5 years, amounting to approximately 0.8 million deaths per year. Currently there is a dearth of literature exploring the effectiveness of antibiotics for diarrhea due to Cholera, Shigella and cryptosporidiosis in children. Methods We reviewed the literature reporting the effect of antibiotics for the treatment of diarrhea due to Cholera, Shigella and Cryptosporidium in children under five years. We used a standardized abstraction and grading format and performed meta-analyses to determine the effect of the treatment with various antibiotics on mortality and rates of clinical and bacteriological/parasitological failure. The CHERG Standard Rules were applied to determine the final effect of treatment with antibiotics on diarrhea morbidity and mortality. Results For Cholera; the evidence was weak to recommend any effect on mortality. For Shigella; there was no data on mortality; either all-cause or cause specific, hence we used clinical failure rates as a proxy for Shigella deaths and propose that treatment of Shigella dysentery with antibiotics can result in a 82% reduction in diarrhea mortality due to Shigella. For cryptosporidiosis; there was data on all-cause mortality but the evidence was weak hence we used clinical failure rates as a proxy for mortality to estimate that antimicrobial treatment of diarrhea due to cryptosporidiosis can result in a 54% reduction in mortality. Conclusions There is evidence to recommend antibiotic use for reduction of morbidity and mortality due to Cholera, Shigella and Cryptosporidium. We recommend that more clinical trials should be conducted to evaluate the efficacy and safety of first- and second- line drugs currently in use for treatment for diarrhea and dysentery in both developing and developed countries. PMID:24564492

  9. Occurrence of Cryptosporidium spp. and Giardia spp. in a public water-treatment system, Paraná, Southern Brazil.

    PubMed

    Almeida, Jonatas Campos; Martins, Felippe Danyel Cardoso; Ferreira Neto, José Maurício; Santos, Maíra Moreira Dos; Garcia, João Luis; Navarro, Italmar Teodorico; Kuroda, Emília Kiyomi; Freire, Roberta Lemos

    2015-01-01

    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. PMID:26291147

  10. Cryptosporidium: Treatment

    MedlinePlus

    ... Healthy Water Healthy Swimming/Recreational Water Global Water, Sanitation, & Hygiene (WASH) Other Uses of Water Water-related ... Healthy Water Healthy Swimming/Recreational Water Global Water, Sanitation, & Hygiene (WASH) Other Uses of Water Water-related ...

  11. Removal Efficiencies and Attachment Coefficients for Cryptosporidium in Sandy Alluvial Riverbank Sediment

    EPA Science Inventory

    Riverbank filtration has been shown to be effective at removing viable Cryptosporidium parvum oocysts and, therefore, drinking water systems that employ riverbank filtration may receive additional treatment credits beyond that which they can obtain using traditional engineering a...

  12. Human infective potential of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in urban wastewater treatment plant effluents.

    PubMed

    Ma, Jiawen; Feng, Yaoyu; Hu, Yue; Villegas, Eric N; Xiao, Lihua

    2016-06-01

    Cryptosporidiosis, giardiasis, and microsporidiosis are important waterborne diseases. In the standard for wastewater treatment plant (WWTP) effluents in China and other countries, the fecal coliform count is the only microbial indicator, raising concerns about the potential for pathogen transmission through WWTP effluent reuse. In this study, we collected 50 effluent samples (30 L/sample) from three municipal WWTPs in Shanghai, China, and analyzed for Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi by microscopy and/or polymerase chain reaction (PCR). Moreover, propidium monoazide (PMA)-PCR was used to assess the viability of oocysts/cysts. The microscopy and PCR-positive rates for Cryptosporidium spp. were 62% and 40%, respectively. The occurrence rates of G. duodenalis were 96% by microscopy and 92-100% by PCR analysis of three genetic loci. Furthermore, E. bieneusi was detected in 70% (35/50) of samples by PCR. Altogether, 10 Cryptosporidium species or genotypes, two G. duodenalis genotypes, and 11 E. bieneusi genotypes were found, most of which were human-pathogenic. The chlorine dioxide disinfection employed in WWTP1 and WWTP3 failed to inactivate the residual pathogens; 93% of the samples from WWTP1 and 83% from WWTP3 did not meet the national standard on fecal coliform levels. Thus, urban WWTP effluents often contain residual waterborne human pathogens. PMID:27280607

  13. Molecular Characterization of Cryptosporidium spp. Isolated From Immunocompromised Patients and Children

    PubMed Central

    Rafiei, Abdollah; Rashno, Zahra; Samarbafzadeh, Alireza; Khademvatan, Shahram

    2014-01-01

    Background: Cryptosporidium is known to be one of the most important causes of diarrhea in children and immunocompromised patients. Genotype characterization of Cryptosporidium species in each region would help in the treatment of this disease, as well as to locate the source of infection and to prevent the disease. Objectives: This current research was conducted in order to analyze the molecular characterization of isolated Cryptosporidium spp. in the Southwest of Iran. Materials and Methods: In this survey, 390 fecal samples were collected from immunocompromised individuals and children under five-years-of-age. Parasitic infection was evaluated using wet mount preparation, formalin ether, a modified acid fast staining method and microscopic examination. Finally, a PCR-RFLP assay was performed on the extracted DNA collected from fecal samples that were positive for Cryptosporidium by the acid fast method. Results: Among the 390 fecal samples, 16 cases (4.1%) were infected with Cryptosporidium. Molecular and genotype characterization found the following protozoan species; 11 Cryptosporidium parvum (68.8%), 4 C. hominis (25%), and one case of C. meleagridis (6.2%). Conclusions: The present study emphasized the public health importance of Cryptosporidium spp. in the study area. In addition, it seems that zoonotic species are the most important causes of infection in the region. As far as we are aware this the first report of a C. meleagridis infection in Iran. PMID:25147696

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

    PubMed

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

    2016-08-15

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

  15. Elimination of viruses, phages, bacteria and Cryptosporidium by a new generation Aquaguard point-of-use water treatment unit.

    PubMed

    Grabow, W O; Clay, C G; Dhaliwal, W; Vrey, M A; Müller, E E

    1999-09-01

    The elimination of human viruses, phages, bacteria and Cryptosporidium oocysts by a new generation commercial Aquaguard purifier for the domestic treatment of drinking water, has been evaluated. The unit basically consists of a candle prefilter, activated carbon filter and ultraviolet irradiation compartment. Drinking water seeded with selected laboratory test strains of resistant micro-organisms was passed through the unit. Similar tests were carried out with sewage-contaminated river water and secondary treated waste water containing naturally occurring organisms. Test procedures were based on internationally accepted principles for the evaluation of point-of-use water treatment units, including a standard test protocol of the United States Environmental Protection Agency. Reduction in numbers of seeded test organisms at several log levels higher than those expected in water for which the unit is intended, was determined by the cultivation of viable organisms. In the case of seeded viruses and Cryptosporidium parvum oocysts the qualitative absence of nucleic acid was determined by the reverse transcriptase polymerase chain reaction (RT-PCR). At the design flow rate of one litre per minute, numbers of polio, hepatitis A, adeno types 2 and 41, rota SA11, human rota and astro viruses, as well as somatic and MS2 coliphages, and Escherichia coli, Streptococcus faecalis, Clostridium perfringens, total coliform bacteria, enterococci, heterotrophic bacteria and C. parvum oocysts, were reduced by more than 99.999% in all waters tested. This efficiency conforms to specifications for such units. The quality of the treated water was well within microbiological limits of international specifications for drinking water. PMID:10546330

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

    PubMed

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

    2007-12-01

    Safe drinking water is a top priority in preventing disease outbreaks and is of general concern to everyone. This study examines the occurrence of Cryptosporidium and Giardia in Hungarian drinking water supplies for the first time. A total of 76 raw and drinking water samples were examined using the U.S. EPA Method 1623. From these 15 of 34 (48.4%) raw water samples tested positive for Giardia and 7 (26.6%) for Cryptosporidium. Twelve of 45 (26.7%) drinking water samples were positive for Giardia and 6 (13.3%) for Cryptosporidium. Overall, Giardia cysts and/or Cryptosporidium oocysts were detected in 48% of the raw water samples and 35% of the drinking water samples. The highest levels in drinking water were found to be 3 oocysts/100 litres of Cryptosporidium and 63.6 cysts/100 litres for Giardia, enough to cause giardiasis. The highest levels in raw water were 1,030 cysts/100 litres for Giardia and 50 oocysts/100 litres for Cryptosporidium and higher oocyst densities were associated with source water receiving effluents from sewage treatment plants or originating from a forest environment. In addition to this monitoring, riverbank filtrated water and raw water from the River Danube in Budapest were monitored in order to ascertain protozoan removal efficiency of riverbank filtration (RBF). A total of 157 samples, including 87 samples from the River Danube and 70 samples post RBF, were examined. Cryptosporidium and Giardia were detected regularly in the river water but never in riverbank filtered water suggesting the effectiveness of RBF as a purification method. The occurrence of Cryptosporidium oocysts and Giardia cysts in the investigated water supplies may require the water utilities and water authorities in Hungary to apply additional monitoring and treatment and/or watershed controls. PMID:17878568

  17. CRYPTOSPORIDIUM INACTIVATION AND REMOVAL RESEARCH

    EPA Science Inventory

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

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

    EPA Science Inventory

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

  19. 40 CFR 141.715 - Microbial toolbox options for meeting Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced.../intake management No prescribed credit. Systems may conduct simultaneous monitoring for treatment bin classification at alternative intake locations or under alternative intake management strategies....

  20. 40 CFR 141.715 - Microbial toolbox options for meeting Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced.../intake management No prescribed credit. Systems may conduct simultaneous monitoring for treatment bin classification at alternative intake locations or under alternative intake management strategies....

  1. 40 CFR 141.715 - Microbial toolbox options for meeting Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced.../intake management No prescribed credit. Systems may conduct simultaneous monitoring for treatment bin classification at alternative intake locations or under alternative intake management strategies....

  2. 40 CFR 141.715 - Microbial toolbox options for meeting Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced.../intake management No prescribed credit. Systems may conduct simultaneous monitoring for treatment bin classification at alternative intake locations or under alternative intake management strategies....

  3. 40 CFR 141.715 - Microbial toolbox options for meeting Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced.../intake management No prescribed credit. Systems may conduct simultaneous monitoring for treatment bin classification at alternative intake locations or under alternative intake management strategies....

  4. Recent additions in the treatment of cough

    PubMed Central

    Gibson, Peter G.

    2014-01-01

    Background Worldwide, cough is regarded as a challenging clinical problem due to its frequency and often limited therapeutic options. Chronic cough that remains refractory to usual medical treatment causes significant quality of life impairment in people with this problem. Methods We have examined current evidence on recent additions in the treatment of cough, specifically treatment of refractory chronic cough with speech pathology and gabapentin. Relevant randomised control trials, reviews and case reports were identified through a PubMed and SCOPUS search of English-language literature referring to these concepts over the last eight years. Summary Of the one hundred and two articles comprising this review the majority investigated the role of the transient receptor potential (TRP) receptors TRP Vanilloid 1 (TRPV1) and TRPA1 in cough and the potential of TRP antagonists as effective anti-tussives. However, these have only been tested in the laboratory and therefore their clinical effectiveness is unknown. Behavioural treatments such as speech pathology have gained momentum and this was evident in the increasing number of articles investigating its positive effect on cough. Investigation on the effectiveness of neuromodulating medications in the treatment of cough have been supported primarily through case series reports and prospective reviews however; their use (particularly gabapentin) has been significantly advanced through recently conducted randomised controlled trials. Conclusions Recent additions in the treatment of chronic cough have been significant as they consider cough to have a unifying diagnosis of cough hypersensitivity with or without the presence of a neuropathic basis. Primarily, effective treatments for chronic cough target these areas and include behavioural treatment such as speech pathology and pharmaceutical treatment with neuromodulating medications such as gabapentin. PMID:25383209

  5. The Cryptosporidium parvum Kinome

    PubMed Central

    2011-01-01

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

  6. Cryptosporidium (Crypto)

    MedlinePlus

    ... Symptoms Signs, symptoms, and outcomes... Treatment Medication... Pathogen & Environment Stages of parasite development... Information for Specific Groups Public Immunocompromised Persons Public ...

  7. Parasitic infection among HIV/AIDS patients at Bela-Bela clinic, Limpopo province, South Africa with special reference to Cryptosporidium.

    PubMed

    Samie, Amidou; Makuwa, Stanley; Mtshali, Sibusiso; Potgieter, Natasha; Thekisoe, Oriel; Mbati, Peter; Bessong, Pascal O

    2014-07-01

    Intestinal parasitic organisms are common pathogens among HIV patients worldwide and have been known to cause severe and life-threatening diarrhea in such subjects. In the present study, the prevalence of Cryptosporidium spp and other intestinal parasites in stool samples from 151 HIV/AIDS patients attending a HIV treatment center in South Africa was determined using' standard parasitological methods, as well as molecular methods including PCR and quantitative PCR for confirmation of Cryptosporidium spp. In addition, the loop-mediated isothermal amplification (LAMP) method was evaluated for detection of Cryptosporidium spp in 24 stool samples. Standard parasitological methods indicated that Cryptospo- ridium spp (26.5%), Entamoeba spp (26.5%) and Giardia lamblia (13%) were the most common protozoan parasites, while Ascaris lumbricoides (8%), Schistosoma mansoni (6%) and Trichuris trichiura (4.6%) were the most commonly found helminths. PCR, quantitative PCR and LAMP methods identified Cryptosporidium spp in 28% (30/106), 35% (53/151) and 58% (14/24) of the stool samples, respectively. Multiple infections (34%) were commonly found in the study population. Females above 45 years had the highest Cryptosporidium prevalence (58%). Prevention measures must be implemented in order to curb the negative impact of Cryptosporidium-causing diarrhea among HIV/AIDS patients in this region as well as other parasitic infections identified in this study. PMID:25427345

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

    PubMed

    Spanakos, Gregory; Biba, Anastasia; Mavridou, Athena; Karanis, Panagiotis

    2015-05-01

    Here, we present the first time findings regarding the occurrence of Cryptosporidium and Giardia in sewage waters and the first molecular characterization of Cryptosporidium species in Greece. Biological treatment plants from three regions in Greece have been investigated. The detection of Cryptosporidium oocysts was by modified Ziehl-Neelsen acid fast (MZN-AF) and by immunofluorescence microscopy (IFT) for Cryptosporidium and Giardia (oo)cysts, whereas nested PCR based on the SSU rDNA assay was used for molecular detection of Cryptosporidium followed by sequencing for the genetic characterization of the species. In total, 73 samples (37 raw sewage samples and 38 of treated water samples) were collected and analyzed. Of the 73 water samples, 4 samples were Cryptosporidium-positive by IFT and staining, 12 samples were Cryptosporidium-positive by nested PCR; 9 samples were Giardia-positive by IFT. We showed that Cryptosporidium cysts are found both in the input and the discharge of the biological treatment plants. Molecular characterization of Cryptosporidium based on the small subunit ribosomal DNA gene resulted in the determination of Cryptosporidium parvum and Cryptosporidium muris Greek isolates. This is the first report of Cryptosporidium and Giardia occurrence in wastewaters and the first molecular identification of Cryptosporidium species in Greek environments. As the treated water is used for irrigation, or it is discharged into the sea, our findings indicate that biological treatment facilities constitute a possible risk for public health because the related species are prevalent in humans; the results invite for further epidemiological investigations to evaluate the real public health risk in Greece. PMID:25687523

  9. Cryptosporidium (Crypto) Disease

    MedlinePlus

    ... Cryptosporidium Tracking in the United States CryptoNet Healthy Water Links Healthy Water Healthy Swimming/Recreational Water Global ... you requested has moved to Illness & Symptoms. Healthy Water Links Healthy Water Healthy Swimming/Recreational Water Global ...

  10. Cryptosporidium: Prevention - Immunocompromised Persons

    MedlinePlus

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

  11. Cryptosporidium, organism (image)

    MedlinePlus

    Cryptosporidium is a protozoan parasite found in contaminated water. It has been increasingly recognized as the cause of outbreaks of diarrhea when water supplies become contaminated. In normal individuals, it is a ...

  12. PAIRED CITY CRYPTOSPORIDIUM SEROSURVEY

    EPA Science Inventory

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

  13. [1N,12N]Bis(Ethyl)-cis-6,7-Dehydrospermine: a New Drug for Treatment and Prevention of Cryptosporidium parvum Infection of Mice Deficient in T-Cell Receptor Alpha

    PubMed Central

    Waters, W. R.; Frydman, B.; Marton, L. J.; Valasinas, A.; Reddy, V. K.; Harp, J. A.; Wannemuehler, M. J.; Yarlett, N.

    2000-01-01

    Cryptosporidium parvum infection of T-cell receptor alpha (TCR-α)-deficient mice results in a persistent infection. In this study, treatment with a polyamine analogue (SL-11047) prevented C. parvum infection in suckling TCR-α-deficient mice and cleared an existing infection in older mice. Treatment with putrescine, while capable of preventing infection, did not clear C. parvum from previously infected mice. These findings provide further evidence that polyamine metabolic pathways are targets for new anticryptosporidial chemotherapeutic agents. PMID:10991882

  14. Detection of Cryptosporidium sp. in two new seal species, Phoca vitulina and Cystophora cristata, and a novel Cryptosporidium genotype in a third seal species, Pagophilus groenlandicus, from the Gulf of Maine.

    PubMed

    Bass, A L; Wallace, C C; Yund, P O; Ford, T E

    2012-04-01

    Data on the geographic distribution and host specificity of Cryptosporidium spp. are critical for developing an understanding of likely transmission patterns in nature. During a molecular-based survey of fecal samples from 293 terrestrial and aquatic animals in Maine, USA, we detected Cryptosporidium sp. in 11 harbor seals (Phoca vitulina), 1 hooded seal (Cystophora cristata), and 1 harp seal (Pagophilus groenlandicus). None of the terrestrial or freshwater mammal fecal samples or bird samples tested positive for Cryptosporidium sp. However, the sequencing results of the small subunit (ssu) rRNA gene indicate that the seals were infected with an undescribed species of Cryptosporidium , previously isolated only from ringed seals (Phoca hispida) in northern Quebec, Canada. In addition, the Cryptosporidium sp. detected in the harp seal is significantly different from the previously observed Cryptosporidium sp. in other seals. We confirmed the genetic distinctiveness of this Cryptosporidium genotype and the identity of the other Cryptosporidium sp. seal ssu rRNA sequences by using data from the 70-kDa heat shock protein gene. Based on phylogenetic reconstructions of both genes, it seems that either Cryptosporidium canis or C. felis are sister species to the seal associated Cryptosporidium spp. Our findings extend the range of " Cryptosporidium sp. seal" well south of the 55th parallel, add other species to the list of seals affected by Cryptosporidium sp., and highlight the presence of unrecognized population and potentially species level variation in Cryptosporidium. PMID:22017467

  15. Cryptosporidium Pathogenicity and Virulence

    PubMed Central

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

    2013-01-01

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

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

    PubMed

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

    2012-10-15

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

  17. Global modelling of Cryptosporidium in surface water

    NASA Astrophysics Data System (ADS)

    Vermeulen, Lucie; Hofstra, Nynke

    2016-04-01

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

  18. Cost-effective alternatives for mitigating Cryptosporidium risk in drinking water and enhancing ecosystem services

    NASA Astrophysics Data System (ADS)

    Bryan, B. A.; Kandulu, J. M.

    2009-08-01

    Under the multibarrier paradigm, water quality management barriers that mitigate risk to consumers are required at multiple points from the catchment to the tap. We present a cost-effectiveness analysis of 13 catchment- and treatment-based management alternatives for mitigating Cryptosporidium risk in the Myponga water supply catchment, South Australia. A broad range of costs and benefits are identified and valued, including setup, operation and maintenance, and opportunity costs, and benefits for ecosystem services including water quality, biodiversity, carbon sequestration, and farm production services. The results suggest that the cost-effectiveness of investment in water quality management can be substantially enhanced by considering the costs of management and the benefits for ecosystem services, in addition to Cryptosporidium removal effectiveness. Cost-effectiveness of investment in management alternatives is dependent upon the desired level of Cryptosporidium removal effectiveness by both the catchment and treatment barriers. The combination of a spatially targeted 25% restriction in water course access of nondairy cattle and treatment by enhanced coagulation provides the most (net) cost-effective Cryptosporidium risk mitigation strategy. This combination may achieve 0.614 log removal at a net cost of A0.7 million and (net) cost-effectiveness of A1.14 million per log removal. Additional risk mitigation can be achieved through the addition of ultraviolet irradiation treatment, higher levels of water course access restriction for cattle, and the adoption of dung beetles in the catchment. Economic valuation of a range of costs and benefits of management priorities can support cost-effective water quality management investment decisions and inform elements of policy design such as cost-sharing arrangements and spatial targeting.

  19. Cryptosporidium and Cryptosporidiosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This 2nd edition of the book “Cryptosporidium and Cryptosporidiosis” has been greatly revised and expanded. Included are the following chapters and subject areas. Chapter 1 discusses general biological issues. It traces the history of discovery of the genus and species, updates the taxonomy, describ...

  20. Additional Treatments Offer Little Benefit for Pancreatic Cancer: Study

    MedlinePlus

    ... 158633.html Additional Treatments Offer Little Benefit for Pancreatic Cancer: Study Neither extra chemotherapy drug nor add-on ... 2016 (HealthDay News) -- Additional treatments for locally advanced pancreatic cancer don't appear to boost survival, a new ...

  1. Giardia and Cryptosporidium spp. dissemination during wastewater treatment and comparative detection via immunofluorescence assay (IFA), nested polymerase chain reaction (nested PCR) and loop mediated isothermal amplification (LAMP).

    PubMed

    Gallas-Lindemann, Carmen; Sotiriadou, Isaia; Plutzer, Judit; Noack, Michael J; Mahmoudi, Mohammad Reza; Karanis, Panagiotis

    2016-06-01

    Environmental water samples from the Lower Rhine area in Germany were investigated via immunofluorescence assays (IFAs), nested polymerase chain reaction (nested PCR) and loop-mediated isothermal amplification (LAMP) to detect the presence of Giardia spp. (n=185) and Cryptosporidium spp. (n=227). The samples were concentrated through filtration or flocculation, and oocysts were purified via centrifugation through a sucrose density gradient. For all samples, IFA was performed first, followed by DNA extraction for the nested PCR and LAMP assays. Giardia cysts were detected in 105 samples (56.8%) by IFA, 62 samples (33.5%) by nested PCR and 79 samples (42.7%) by LAMP. Cryptosporidium spp. were detected in 69 samples (30.4%) by IFA, 95 samples (41.9%) by nested PCR and 99 samples (43.6%) by LAMP. According to these results, the three detection methods are complementary for monitoring Giardia and Cryptosporidium in environmental waters. PMID:26880717

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  3. Giardia cysts and Cryptosporidium oocysts in membrane-filtered municipal wastewater used for irrigation.

    PubMed

    Lonigro, A; Pollice, A; Spinelli, R; Berrilli, F; Di Cave, D; D'Orazi, C; Cavallo, P; Brandonisio, O

    2006-12-01

    A wastewater tertiary treatment system based on membrane ultrafiltration and fed with secondary-treated municipal wastewater was evaluated for its Giardia cyst and Cryptosporidium oocyst removal efficiency. Giardia duodenalis (assemblages A and B) and Cryptosporidium parvum were identified in feed water but were found in filtered water only during occasional failure of the filtration system. PMID:17056696

  4. 10. Water treatment plant, view to S. 1965 addition is ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    10. Water treatment plant, view to S. 1965 addition is in the foreground - Fort Benton Water Treatment Plant, Filtration Plant, Lots 9-13 of Block 7, Fort Benton Original Townsite at Missouri River, Fort Benton, Chouteau County, MT

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

    PubMed

    Cummins, E; Kennedy, R; Cormican, M

    2010-01-15

    Cryptosporidium species are protozoan parasites associated with gastro-intestinal illness. Following a number of high profile outbreaks worldwide, it has emerged as a parasite of major public health concern. A quantitative Monte Carlo simulation model was developed to evaluate the annual risk of infection from Cryptosporidium in tap water in Ireland. The assessment considers the potential initial contamination levels in raw water, oocyst removal and decontamination events following various process stages, including coagulation/flocculation, sedimentation, filtration and disinfection. A number of scenarios were analysed to represent potential risks from public water supplies, group water schemes and private wells. Where surface water is used additional physical and chemical water treatment is important in terms of reducing the risk to consumers. The simulated annual risk of illness for immunocompetent individuals was below 1 x 10(-4) per year (as set by the US EPA) except under extreme contamination events. The risk for immunocompromised individuals was 2-3 orders of magnitude greater for the scenarios analysed. The model indicates a reduced risk of infection from tap water that has undergone microfiltration, as this treatment is more robust in the event of high contamination loads. The sensitivity analysis highlighted the importance of watershed protection and the importance of adequate coagulation/flocculation in conventional treatment. The frequency of failure of the treatment process is the most important parameter influencing human risk in conventional treatment. The model developed in this study may be useful for local authorities, government agencies and other stakeholders to evaluate the likely risk of infection given some basic input data on source water and treatment processes used. PMID:19945145

  6. 40 CFR 141.211 - Special notice for repeated failure to conduct monitoring of the source water for Cryptosporidium...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

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

  7. 40 CFR 141.211 - Special notice for repeated failure to conduct monitoring of the source water for Cryptosporidium...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

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

  8. 40 CFR 141.211 - Special notice for repeated failure to conduct monitoring of the source water for Cryptosporidium...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

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

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

    PubMed

    Arnone, Russell D; Walling, Joyce Perdek

    2006-06-01

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

  10. 39 CFR 3050.41 - Treatment of additional financial reports.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 39 Postal Service 1 2010-07-01 2010-07-01 false Treatment of additional financial reports. 3050.41 Section 3050.41 Postal Service POSTAL REGULATORY COMMISSION PERSONNEL PERIODIC REPORTING § 3050.41 Treatment of additional financial reports. (a) For purposes of the reports required by § 3050.40(a)(2),...

  11. Cryptosporidium source tracking in the Potomac River watershed.

    PubMed

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

    2008-11-01

    To better characterize Cryptosporidium in the Potomac River watershed, a PCR-based genotyping tool was used to analyze 64 base flow and 28 storm flow samples from five sites in the watershed. These sites included two water treatment plant intakes, as well as three upstream sites, each associated with a different type of land use. The uses, including urban wastewater, agricultural (cattle) wastewater, and wildlife, posed different risks in terms of the potential contribution of Cryptosporidium oocysts to the source water. Cryptosporidium was detected in 27 base flow water samples and 23 storm flow water samples. The most frequently detected species was C. andersoni (detected in 41 samples), while 14 other species or genotypes, almost all wildlife associated, were occasionally detected. The two common human-pathogenic species, C. hominis and C. parvum, were not detected. Although C. andersoni was common at all four sites influenced by agriculture, it was largely absent at the urban wastewater site. There were very few positive samples as determined by Environmental Protection Agency method 1623 at any site; only 8 of 90 samples analyzed (9%) were positive for Cryptosporidium as determined by microscopy. The genotyping results suggest that many of the Cryptosporidium oocysts in the water treatment plant source waters were from old calves and adult cattle and might not pose a significant risk to human health. PMID:18776033

  12. Human infective potential of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in urban wastewater treatment plant effluents

    EPA Science Inventory

    Cryptosporidiosis, giardiasis, and microsporidiosis are important waterborne diseases. In the standard for wastewater treatment plant (WWTP) effluents in China and other countries, fecal coliform is the only microbial indicator, raising concerns about the potential for pathogen t...

  13. Cryptosporidium Source Tracking in the Potomac River Watershed

    EPA Science Inventory

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

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

    EPA Science Inventory

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

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

    EPA Science Inventory

    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...

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

    EPA Science Inventory

    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...

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

    PubMed Central

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

    1991-01-01

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

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

    PubMed

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

    2015-12-01

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

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

    PubMed Central

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

    2015-01-01

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

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

    PubMed

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

    2015-06-01

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

  1. Cryptosporidium parvum Infection Following Contact with Livestock

    PubMed Central

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

    2016-01-01

    Context: Scours, or calf diarrhea, is an infectious gastrointestinal disease commonly found in the calves of dairy farms. It primarily presents with diarrhea that can be life threatening to the animal and is also contagious and threatening to the other livestock. Cryptosporidium is one of the major causes of scours and can be transmitted to humans via fecal-oral route, resulting in diarrheal illnesses. Cryptosporidiosis infection usually occurs as a waterborne outbreak with the potential to affect many people at once. Case Report: We report a case of a 24-year-old female farmer who presented to the emergency department with diarrhea after taking care of ill cattle with similar symptoms. Fecal cultures were positive for Cryptosporidium parvum. Given the patient was immunocompetent, no further treatment was warranted. Conclusion: Confirmed cases should be reported, however, treatment is only recommended in children and immunocompromised adults. Clinicians should educate patients on the importance of proper hygiene and handling techniques in order to decrease transmission and recurrence of the protozoan infection. PMID:27583243

  2. Host responses to Cryptosporidium infection.

    PubMed

    Gookin, Jody L; Nordone, Shila K; Argenzio, Robert A

    2002-01-01

    Cryptosporidium is a clinically and economically important infection whose pathogenic effect begins with colonization of the intestinal epithelium. Despite intensive efforts, a consistently effective therapy for the infection has yet to be identified. Morbidity and mortality results from ongoing loss of absorptive epithelium, which leads to villous atrophy and malabsorption and release of inflammatory mediators that stimulate electrolyte secretion and diarrhea. With further clarification of the mechanisms underlying enterocyte malfunction in Cryptosporidium infection, it should be possible to design rational nutritional and pharmacologic therapies to enhance nutrient and water absorption, promote the clearance of infected enterocytes, and restore normal villus architecture and mucosal barrier function. PMID:11822801

  3. Occurrence of Cryptosporidium suis and Cryptosporidium scrofarum on commercial swine farms in the Czech Republic and its associations with age and husbandry practices.

    PubMed

    Němejc, Karel; Sak, Bohumil; Květoňová, Dana; Kernerová, Naděžda; Rost, Michael; Cama, Vitaliano A; Kváč, Martin

    2013-03-01

    From 2009 to 2011, the occurrence of Cryptosporidium spp. was investigated on 22 farms in the Czech Republic. A total of 1,620 individual faecal samples of pigs of all age categories (pre-weaned, starters, pre-growers, growers, and sows) were evaluated for presence of Cryptosporidium spp. by standard microscopy and molecular tools. Genotyping was done through PCR amplification and characterization of the SSU rRNA (species-specific protocols) and GP60 loci. Cryptosporidium spp. was found on 16 of 22 farms with a range 0.9-71.4 %. Overall, 194 (12 %) specimens were positive by microscopy and 353 (21.8 %) by PCR. While RFLP and direct sequencing of the PCR-amplified products showed presence of Cryptosporidium suis (142), Cryptosporidium scrofarum (195), Cryptosporidium muris (3) and 13 samples had mixed infections with C. suis and C. scrofarum, species-specific molecular tools identified C. suis (224), C. scrofarum (208), Cryptosporidium parvum subtype IIa A16G1R1b (1), and C. muris (3). In addition, a total of 82 pigs had concurrent infections with C. suis and C. scrofarum. The analysis by age showed that C. suis was primarily detected among pre-weaned, whereas C. scrofarum was mostly detected among starters, especially those weaned at a younger age. Moreover, C. scrofarum never has been detected in animals younger than 6 weeks of age. Also, piglets weaned at 3 weeks of age were twice more likely to be infected with C. scrofarum than piglets weaned at an older age. Pigs raised on straw bedding were more likely to have Cryptosporidium than pigs raised on slats/slurry systems. The infections with different species were not associated with loose faeces or intensity of oocyst shedding, even when comparing different age groups. PMID:23271566

  4. Occurrence of Cryptosporidium suis and Cryptosporidium scrofarum on commercial swine farms in the Czech Republic and its associations with age and husbandry practices

    PubMed Central

    Němejc, Karel; Sak, Bohumil; Květoňová, Dana; Kernerová, Naděžda; Rost, Michael; Cama, Vitaliano A.; Kváč, Martin

    2015-01-01

    From 2009 to 2011, the occurrence of Cryptosporidium spp. was investigated on 22 farms in the Czech Republic. A total of 1,620 individual faecal samples of pigs of all age categories (pre-weaned, starters, pre-growers, growers, and sows) were evaluated for presence of Cryptosporidium spp. by standard microscopy and molecular tools. Genotyping was done through PCR amplification and characterization of the SSU rRNA (species-specific protocols) and GP60 loci. Cryptosporidium spp. was found on 16 of 22 farms with a range 0.9–71.4 %. Overall, 194 (12 %) specimens were positive by microscopy and 353 (21.8 %) by PCR. While RFLP and direct sequencing of the PCR-amplified products showed presence of Cryptosporidium suis (142), Cryptosporidium scrofarum (195), Cryptosporidium muris (3) and 13 samples had mixed infections with C. suis and C. scrofarum, species-specific molecular tools identified C. suis (224), C. scrofarum (208), Cryptosporidium parvum subtype IIa A16G1R1b (1), and C. muris (3). In addition, a total of 82 pigs had concurrent infections with C. suis and C. scrofarum. The analysis by age showed that C. suis was primarily detected among pre-weaned, whereas C. scrofarum was mostly detected among starters, especially those weaned at a younger age. Moreover, C. scrofarum never has been detected in animals younger than 6 weeks of age. Also, piglets weaned at 3 weeks of age were twice more likely to be infected with C. scrofarum than piglets weaned at an older age. Pigs raised on straw bedding were more likely to have Cryptosporidium than pigs raised on slats/slurry systems. The infections with different species were not associated with loose faeces or intensity of oocyst shedding, even when comparing different age groups. PMID:23271566

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

    PubMed

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

    2014-01-01

    Continuous exposure to low levels of Cryptosporidium oocysts is associated with production of protective antibodies. We investigated prevalence of antibodies against the 27-kDa Cryptosporidium oocyst antigen among blood donors in 2 areas of Scotland supplied by drinking water from different sources with different filtration standards: Glasgow (not filtered) and Dundee (filtered). During 2006-2009, seroprevalence and risk factor data were collected; this period includes 2007, when enhanced filtration was introduced to the Glasgow supply. A serologic response to the 27-kDa antigen was found for ≈75% of donors in the 2 cohorts combined. Mixed regression modeling indicated a 32% step-change reduction in seroprevalence of antibodies against Cryptosporidium among persons in the Glasgow area, which was associated with introduction of enhanced filtration treatment. Removal of Cryptosporidium oocysts from water reduces the risk for waterborne exposure, sporadic infections, and outbreaks. Paradoxically, however, oocyst removal might lower immunity and increase the risk for infection from other sources. PMID:24377436

  6. The General Biology of Cryptosporidium

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

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

    PubMed

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

    2010-12-01

    The objective of this study was to determine the mean concentration (per litre) of Cryptosporidium oocysts and Giardia cysts in recreational river areas (n = 28), drinking water treatments plants (DWTPs; n = 52) and wastewater treatment plants (WWTPs; n = 50) in Galicia (NW Spain). Water samples from rivers and from the influent (50-100 l) and the treated effluent (100 l) of the water plants were filtered using Filta-Max filters (IDEXX Laboratories, Inc., Westbrook, ME, USA). A total of 232 samples were processed and the (oo)cysts were concentrated, clarified by IMS and then detected by IFAT. The viability was determined by applying fluorogenic vital dye (PI). In the recreational areas, infective forms of Cryptosporidium and Giardia were detected in 16 (57.1%; 1-60 oocysts per litre) and 17 (60.7%; 1-160 cysts per litre) samples, respectively. In the water flowing into the water treatment plants, oocysts were detected in 21 DWTPs (40.4%; 1-13 oocysts per litre) and cysts were observed in 22 DWTPs (42.3%; 1-7 cysts per litre). In the effluents from the treatment plants, Cryptosporidium oocysts and Giardia cysts were identified in 17 DWTPs (32.7%; 1-4 oocysts per litre) and in 19 DWTPs (36.5%; 1-5 cysts per litre), respectively. The highest concentrations of (oo)cysts were found in the WWTPs; specifically, oocysts were detected in 29 (58.0%; 1-80 oocysts per litre) and cysts in 49 (98.0%; 2-14.400 cysts per litre) WWTP effluents. Cryptosporidium and Giardia were detected in 32 (64.0%; 1-120 oocysts per litre) and 48 (96.0%; 2-6.000 cysts per litre) WWTP effluents, respectively. The percentage viability of the (oo)cysts ranged between 90.0% and 95.0%. In all samples analysed. Moreover, it was found that the effluents from coastal WWTPs were discharged directly into the sea, while inland WWTPs were discharged directly into rivers. The concentrations of both enteropathogens detected in effluents from WWTPs therefore represent a significant risk to human and

  8. Fecal Indole as a Biomarker of Susceptibility to Cryptosporidium Infection.

    PubMed

    Chappell, Cynthia L; Darkoh, Charles; Shimmin, Lawrence; Farhana, Naveed; Kim, Do-Kyun; Okhuysen, Pablo C; Hixson, James

    2016-08-01

    Cryptosporidium causes significant diarrhea worldwide, especially among children and immunocompromised individuals, and no effective drug treatment is currently available for those who need it most. In this report, previous volunteer infectivity studies have been extended to examine the association between fecal indole and indole-producing (IP) gut microbiota on the outcome of a Cryptosporidium infection. Fecal indole concentrations (FICs) of 50 subjects and 19 taxa of common gut microbiota, including six IP taxa (11 subjects) were determined in stool samples collected before and after a challenge with Cryptosporidium oocysts. At the baseline, the mean FIC (± the standard deviation) was 1.66 ± 0.80 mM in those who became infected after a challenge versus 3.20 ± 1.23 mM in those who remained uninfected (P = 0.0001). Only 11.1% of the subjects with a FIC of >2.5 mM became infected after a challenge versus 65.2% of the subjects with a FIC of <2.5 mM. In contrast, the FICs of infected subjects at the baseline or during diarrhea were not correlated with infection intensity or disease severity. The relative abundances (percent) of Escherichia coli, Bacillus spp., and Clostridium spp. were greater ≥2.5-fold in volunteers with a baseline FIC of >2.5 mM, while those of Bacteroides pyogenes, B. fragilis, and Akkermansia muciniphila were greater in those with a baseline FIC of <2.5 mM. These data indicate that some IP bacteria, or perhaps indole alone, can influence the ability of Cryptosporidium to establish an infection. Thus, preexisting indole levels in the gut join the oocyst dose and immune status as important factors that determine the outcome of Cryptosporidium exposure. PMID:27245413

  9. Additional Treatment Services in a Cocaine Treatment Study: Level of Services Obtained and Impact on Outcome

    PubMed Central

    Worley, Matthew; Gallop, Robert; Gibbons, Mary Beth Connolly; Ring-Kurtz, Sarah; Present, Julie; Weiss, Roger D.; Crits-Christoph, Paul

    2009-01-01

    The objective of this study was to examine the level of additional treatment services obtained by patients enrolled in the NIDA Cocaine Collaborative Study, a multi-center efficacy trial of four treatments for cocaine dependence, and to determine whether these services impact treatment outcome. Cocaine-dependent patients (N = 487) were recruited at five sites and randomly assigned to six months of one of four psychosocial treatments. Assessments were made at baseline, monthly during treatment, and at follow-ups at 9, 12, 15, and 18 months post-randomization. On average, patients received little or no additional treatment services during active treatment (first 6 months), but the rate of obtaining most services increased during the follow-up phase (month 7 to 18). In general, the treatment groups did not differ in the rates of obtaining non-protocol services. For all treatment groups, patients with greater psychiatric severity received more medical and psychiatric services during active treatment and follow-up. Use of treatment services was unrelated to drug use outcomes during active treatment. However, during the follow-up period, increased use of psychiatric medication, 12-step attendance, and 12-step participation was related to less drug use. The results suggest that during uncontrolled follow-up phases, additional non-protocol services may potentially confound the interpretation of treatment group comparisons in drug use outcomes. PMID:18463998

  10. Censored data treatment using additional information in intelligent medical systems

    NASA Astrophysics Data System (ADS)

    Zenkova, Z. N.

    2015-11-01

    Statistical procedures are a very important and significant part of modern intelligent medical systems. They are used for proceeding, mining and analysis of different types of the data about patients and their diseases; help to make various decisions, regarding the diagnosis, treatment, medication or surgery, etc. In many cases the data can be censored or incomplete. It is a well-known fact that censorship considerably reduces the efficiency of statistical procedures. In this paper the author makes a brief review of the approaches which allow improvement of the procedures using additional information, and describes a modified estimation of an unknown cumulative distribution function involving additional information about a quantile which is known exactly. The additional information is used by applying a projection of a classical estimator to a set of estimators with certain properties. The Kaplan-Meier estimator is considered as an estimator of the unknown cumulative distribution function, the properties of the modified estimator are investigated for a case of a single right censorship by means of simulations.

  11. RNA Extraction Methods for Real-Time PCR and Microarray Analyses of Cryptosporidium and Toxoplasma gondii Oocysts - 2nd Presentation

    EPA Science Inventory

    The ability of infectious oocyst forms of Toxoplasma gondii and Cryptosporidium spp. to resist disinfection treatments and cause disease may have significant public health implications. Currently, little is known about oocyst-specific factors involved during host cell invasion pr...

  12. RNA Extraction Methods for Reverse Transcriptase Real-Time PCR and Microarray Analysis of Cryptosporidium and Toxoplasma gondii Oocysts

    EPA Science Inventory

    The ability of infectious oocyst forms of Toxoplasma gondii and Cryptosporidium spp. to resist disinfection treatments and cause disease may have significant public health implications. Currently, little is known about oocyst-specific factors involved during host cell invasion p...

  13. Molecular identification of Cryptosporidium spp. in seagulls, pigeons, dogs, and cats in Thailand

    PubMed Central

    Koompapong, Khuanchai; Mori, Hirotake; Thammasonthijarern, Nipa; Prasertbun, Rapeepun; Pintong, Ai-rada; Popruk, Supaluk; Rojekittikhun, Wichit; Chaisiri, Kittipong; Sukthana, Yaowalark; Mahittikorn, Aongart

    2014-01-01

    Zoonotic Cryptosporidium spp., particularly C. meleagridis, C. canis, and C. felis, are enteric protozoa responsible for major public health concerns around the world. To determine the spread of this parasite in Thailand, we conducted molecular identification of Cryptosporidium spp. from animal samples around the country, by collecting and investigating the feces of seagulls (Chroicocephalus brunnicephalus and Chroicocephalus ridibundus), domestic pigeons (Columba livia domestica), dogs, and cats. Seagull and pigeon samples were collected at the seaside and on the riverside to evaluate their potential for waterborne transmission. Ten pigeon samples were combined into one set, and a total of seven sets were collected. Seventy seagull samples were combined into one set, and a total of 13 sets were collected. In addition, 111 dog samples were collected from cattle farms, and 95 dog and 80 cat samples were collected from a temple. We identified C. meleagridis in pigeons, Cryptosporidium avian genotype III in seagulls, C. canis in dogs, and C. felis in cats. In the temple, the prevalence was 2.1% (2/95) for dogs and 2.5% (2/80) for cats. No Cryptosporidium was found in dog samples from cattle farms. These are the first findings of C. meleagridis in domestic pigeons, and Cryptosporidium avian genotype III in seagulls. Our study invites further molecular epidemiological investigations of Cryptosporidium in these animals and their environment to evaluate the public health risk in Thailand. PMID:25297887

  14. Age-related detection and molecular characterization of Cryptosporidium suis and Cryptosporidium scrofarum in pre- and post-weaned piglets and adult pigs in Japan.

    PubMed

    Yui, Takeshi; Nakajima, Toshiyuki; Yamamoto, Norishige; Kon, Marina; Abe, Niichiro; Matsubayashi, Makoto; Shibahara, Tomoyuki

    2014-01-01

    We investigated the distribution of Cryptosporidium in pigs in Japan by immunofluorescence staining of fecal samples and characterization of isolates by multilocus sequencing. The 344 animals sampled on eight farms included pre-weaned piglets (<1 month old; n = 55), weaned piglets (1-2 months old; n = 65), finished pigs (2-4 months old, n = 105) and of 4-6 months old (n = 67), sows (n = 36), and boars (n = 16). Average prevalence of Cryptosporidium on farms was 32.6%, ranging from 4.9 to 58.1%, decreasing with animal age (prevalences of <1 month old, 1-2 months old, 2-4 months old, 4-6 months old, sows, and boars were 27.3, 47.7, 41.9, 22.4, 11.1, 18.8%, respectively). Piglets (<1 and 1-2 months old) showing signs of diarrhea shed relatively more oocysts (5.28 in average log scale of oocysts per gram) in feces than piglets with normal or loose stools (those of 4.90). Thirty seven successful sequencing of the 18S ribosomal RNA gene among 62 examined samples revealed that all of the identified isolates were Cryptosporidium suis or Cryptosporidium scrofarum, which are generally specific to pigs, and that other species, such as zoonotic Cryptosporidium parvum, were absent. Interestingly, C. suis was frequently found in piglets younger than 2 months old, while C. scrofarum infection was more prevalent in older pigs which also showed increased prevalence of mixed C. suis and C. scrofarum infections. Sequencing of actin gene loci revealed the existence of variants of both Cryptosporidium species in pigs in Japan. Although the number of pigs examined in this study was relatively low, our results suggest that Cryptosporidium infection is widespread among pigs in Japan. In addition, the possibility of age-related specificity and pathogenicity in pig infections is also suggested. PMID:24189974

  15. CRYPTOSPORIDIUM LOG-INACTIVATION WITH OZONE USING EFFLUENT CT 10, GEOMETRIC MEAN CT 10 EXTENDED INTEGRATED CT 10 AND EXTENDED-CSTR CALCULATIONS

    EPA Science Inventory

    The draft Long Term 2 Enhanced Surface Water Treatment Rule ("LT2ESWTR") contains Cryptosporidium log-inactivation CT tables. Depending on the water temperature, the Cryptosporidium CT values that are listed are 15 to 25 times greater than CT values fo...

  16. Prospects for immunotherapy and vaccines against Cryptosporidium

    PubMed Central

    Mead, Jan R

    2014-01-01

    Cryptosporidium spp is a ubiquitous parasite that has long been recognized as a frequent cause of protozoal diarrhea in humans. While infections in immunocompetent hosts are usually self-limiting, immunocompromised individuals can develop severe, chronic, and life-threatening illness. Vaccine development or immunotherapy that prevents disease or reduces the severity of infection is a relevant option since efficacious drug treatments are lacking. In particular, children in developing countries might benefit the most from a vaccine since cryptosporidiosis in early childhood has been reported to be associated with subsequent impairment in growth, physical fitness, and intellectual capacity. In this review, immunotherapies that have been used clinically are described as well as experimental vaccines and their evaluation in vivo. PMID:24638018

  17. Additives

    NASA Technical Reports Server (NTRS)

    Smalheer, C. V.

    1973-01-01

    The chemistry of lubricant additives is discussed to show what the additives are chemically and what functions they perform in the lubrication of various kinds of equipment. Current theories regarding the mode of action of lubricant additives are presented. The additive groups discussed include the following: (1) detergents and dispersants, (2) corrosion inhibitors, (3) antioxidants, (4) viscosity index improvers, (5) pour point depressants, and (6) antifouling agents.

  18. Genetic Diversity of Cryptosporidium spp. in Captive Reptiles

    PubMed Central

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

    2004-01-01

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

  19. Giardia and Cryptosporidium spp. in filtered drinking water supplies.

    PubMed

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

    1991-09-01

    Giardia and Cryptosporidium levels were determined by using a combined immunofluorescence test for filtered drinking water samples collected from 66 surface water treatment plants in 14 states and 1 Canadian province. Giardia cysts were detected in 17% of the 83 filtered water effluents. Cryptosporidium oocysts, were observed in 27% of the drinking water samples. Overall, cysts or oocysts were found in 39% of the treated effluent samples. Despite the frequent detection of parasites in drinking water, microscopic observations of the cysts and oocysts suggested that most of the organisms were nonviable. Compliance with the filtration criteria outlined by the Surface Water Treatment Rule of the U.S. Environmental Protection Agency did not ensure that treated water was free of cysts and oocysts. The average plant effluent turbidity for sites which were parasite positive was 0.19 nephelometric turbidity units. Of sites that were positive for Giardia or Cryptosporidium spp., 78% would have been able to meet the turbidity regulations of the Surface Water Temperature Rule. Evaluation of the data by using a risk assessment model developed for Giardia spp. showed that 24% of the utilities examined would not meet a 1/10,000 annual risk of Giardia infection. For cold water conditions (0.5 degree C), 46% of the plants would not achieve the 1/10,000 risk level. PMID:1768135

  20. Giardia and Cryptosporidium spp. in filtered drinking water supplies.

    PubMed Central

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

    1991-01-01

    Giardia and Cryptosporidium levels were determined by using a combined immunofluorescence test for filtered drinking water samples collected from 66 surface water treatment plants in 14 states and 1 Canadian province. Giardia cysts were detected in 17% of the 83 filtered water effluents. Cryptosporidium oocysts, were observed in 27% of the drinking water samples. Overall, cysts or oocysts were found in 39% of the treated effluent samples. Despite the frequent detection of parasites in drinking water, microscopic observations of the cysts and oocysts suggested that most of the organisms were nonviable. Compliance with the filtration criteria outlined by the Surface Water Treatment Rule of the U.S. Environmental Protection Agency did not ensure that treated water was free of cysts and oocysts. The average plant effluent turbidity for sites which were parasite positive was 0.19 nephelometric turbidity units. Of sites that were positive for Giardia or Cryptosporidium spp., 78% would have been able to meet the turbidity regulations of the Surface Water Temperature Rule. Evaluation of the data by using a risk assessment model developed for Giardia spp. showed that 24% of the utilities examined would not meet a 1/10,000 annual risk of Giardia infection. For cold water conditions (0.5 degree C), 46% of the plants would not achieve the 1/10,000 risk level. PMID:1768135

  1. Critical processes affecting Cryptosporidium oocyst survival in the environment.

    PubMed

    King, B J; Monis, P T

    2007-03-01

    Cryptosporidium are parasitic protozoans that cause gastrointestinal disease and represent a significant risk to public health. Cryptosporidium oocysts are prevalent in surface waters as a result of human, livestock and native animal faecal contamination. The resistance of oocysts to the concentrations of chlorine and monochloramine used to disinfect potable water increases the risk of waterborne transmission via drinking water. In addition to being resistant to commonly used disinfectants, it is thought that oocysts can persist in the environment and be readily mobilized by precipitation events. This paper will review the critical processes involved in the inactivation or removal of oocysts in the terrestrial and aquatic environments and consider how these processes will respond in the context of climate change. PMID:17096874

  2. Paired city cryptosporidium serosurvey in the southwest USA.

    PubMed Central

    Frost, F. J.; Muller, T.; Craun, G. F.; Calderon, R. L.; Roefer, P. A.

    2001-01-01

    In 1996, serological responses to two Cryptosporidium antigens were determined for 200 Las Vegas (LV), Nevada, and 200 Albuquerque, New Mexico, blood donors to evaluate associations between endemic infections, water exposures, and other risk factors. LV uses chlorinated filtered drinking water from Lake Mead while Albuquerque uses chlorinated ground water. The intensity of serological response to both markers was higher for older donors (P < 0.05). donors who washed food with bottled water (P < 0.05) and donors from LV (P < 0.05). A decreased serological response was not associated with bottled water consumption, nor was an increased response associated with self-reported cryptosporidiosis-like illness or residence in LV at the time of a cryptosporidiosis outbreak 2 years earlier. Although these findings suggest the serological response may be associated with type of tap water and certain foods, additional research is needed to clarifythe role of both food and drinking water in endemic Cryptosporidium infection. PMID:11349981

  3. Cryptosporidium species a "new" human pathogen.

    PubMed Central

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

    1985-01-01

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

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

    PubMed

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

    2014-05-15

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

  5. Cryptosporidium parvum, a potential cause of colic adenocarcinoma

    PubMed Central

    Certad, Gabriela; Ngouanesavanh, Tramy; Guyot, Karine; Gantois, Nausicaa; Chassat, Thierry; Mouray, Anthony; Fleurisse, Laurence; Pinon, Anthony; Cailliez, Jean-Charles; Dei-Cas, Eduardo; Creusy, Colette

    2007-01-01

    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 cryptosporidiosis using SCID mice

  6. Genotyping of Cryptosporidium spp. from free-living wild birds from Brazil.

    PubMed

    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

  7. Distribution and Clinical Manifestations of Cryptosporidium Species and Subtypes in HIV/AIDS Patients in Ethiopia

    PubMed Central

    Adamu, Haileeyesus; Petros, Beyene; Zhang, Guoqing; Kassa, Hailu; Amer, Said; Ye, Jianbin; Feng, Yaoyu; Xiao, Lihua

    2014-01-01

    Background Cryptosporidiosis is an important cause for chronic diarrhea and death in HIV/AIDS patients. Among common Cryptosporidium species in humans, C. parvum is responsible for most zoonotic infections in industrialized nations. Nevertheless, the clinical significance of C. parvum and role of zoonotic transmission in cryptosporidiosis epidemiology in developing countries remain unclear. Methodology/Principal Findings In this cross-sectional study, 520 HIV/AIDS patients were examined for Cryptosporidium presence in stool samples using genotyping and subtyping techniques. Altogether, 140 (26.9%) patients were positive for Cryptosporidium spp. by PCR-RFLP analysis of the small subunit rRNA gene, belonging to C. parvum (92 patients), C. hominis (25 patients), C. viatorum (10 patients), C. felis (5 patients), C. meleagridis (3 patients), C. canis (2 patients), C. xiaoi (2 patients), and mixture of C. parvum and C. hominis (1 patient). Sequence analyses of the 60 kDa glycoprotein gene revealed a high genetic diversity within the 82 C. parvum and 19 C. hominis specimens subtyped, including C. parvum zoonotic subtype families IIa (71) and IId (5) and anthroponotic subtype families IIc (2), IIb (1), IIe (1) and If-like (2), and C. hominis subtype families Id (13), Ie (5), and Ib (1). Overall, Cryptosporidium infection was associated with the occurrence of diarrhea and vomiting. Diarrhea was attributable mostly to C. parvum subtype family IIa and C. hominis, whereas vomiting was largely attributable to C. hominis and rare Cryptosporidium species. Calf contact was identified as a significant risk factor for infection with Cryptosporidium spp., especially C. parvum subtype family IIa. Conclusions/Significance Results of the study indicate that C. parvum is a major cause of cryptosporidiosis in HIV-positive patients and zoonotic transmission is important in cryptosporidiosis epidemiology in Ethiopia. In addition, they confirm that different Cryptosporidium species and

  8. Use of aerobic spores as a surrogate for cryptosporidium oocysts in drinking water supplies.

    PubMed

    Headd, Brendan; Bradford, Scott A

    2016-03-01

    Waterborne illnesses are a growing concern among health and regulatory agencies worldwide. The United States Environmental Protection Agency has established several rules to combat the contamination of water supplies by cryptosporidium oocysts, however, the detection and study of cryptosporidium oocysts is hampered by methodological and financial constraints. As a result, numerous surrogates for cryptosporidium oocysts have been proposed by the scientific community and efforts are underway to evaluate many of the proposed surrogates. The purpose of this review is to evaluate the suitability of aerobic bacterial spores to serve as a surrogate for cryptosporidium oocysts in identifying contaminated drinking waters. To accomplish this we present a comparison of the biology and life cycles of aerobic spores and oocysts and compare their physical properties. An analysis of their surface properties is presented along with a review of the literature in regards to the transport, survival, and prevalence of aerobic spores and oocysts in the saturated subsurface environment. Aerobic spores and oocysts share many commonalities with regard to biology and survivability, and the environmental prevalence and ease of detection make aerobic spores a promising surrogate for cryptosporidium oocysts in surface and groundwater. However, the long-term transport and release of aerobic spores still needs to be further studied, and compared with available oocyst information. In addition, the surface properties and environmental interactions of spores are known to be highly dependent on the spore taxa and purification procedures, and additional research is needed to address these issues in the context of transport. PMID:26734779

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

    PubMed

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

    2012-06-01

    Cryptosporidium and Giardia were characterized in a watershed in southern Ontario, Canada, over a 2½ year period. River samples were collected every two weeks, primarily near a municipal drinking water treatment plant intake. Cryptosporidium and Giardia were frequently detected with an overall occurrence rate of 88 and 97%, respectively. Giardia concentrations were higher than Cryptosporidium, with median values of 80 cysts 100 L(-1) and 12 oocysts 100 L(-1), respectively. Although pathogens rarely show a significant relationship with fecal or water quality indicators, this study determined that Cryptosporidium, but not Giardia, was significantly correlated with Escherichia coli, turbidity and river flow. There was no correlation between the two types of protozoa, and only Giardia showed a seasonal trend with higher concentrations at cold water temperatures. Cryptosporidium genotyping of all samples found that farm animals and wildlife were an important contributor of oocysts in the watershed, and that Cryptosporidium strains/genotypes of medium to high risk for human infection (C. hominis, C. parvum and C. ubiquitum) were detected in 16% of samples. This study was able to identify Cryptosporidium host sources and human health risk, and to identify differences between Cryptosporidium and Giardia occurrence in the watershed. PMID:22717756

  10. Structure of Cryptosporidium IMP dehydrogenase bound to an inhibitor with in vivo antiparasitic activity

    DOE PAGESBeta

    Kim, Youngchang; Makowska-Grzyska, Magdalena; Gorla, Suresh Kumar; Gollapalli, Deviprasad R.; Cuny, Gregory D.; Joachimiak, Andrzej; Hedstrom, Lizbeth

    2015-04-21

    Inosine 5´-monophosphate dehydrogenase (IMPDH) is a promising target for the treatment of Cryptosporidium infections. Here, the structure of C. parvum IMPDH (CpIMPDH) in complex with inosine 5´-monophosphate (IMP) and P131, an inhibitor with in vivo anticryptosporidial activity, is reported. P131 contains two aromatic groups, one of which interacts with the hypoxanthine ring of IMP, while the second interacts with the aromatic ring of a tyrosine in the adjacent subunit. In addition, the amine and NO2 moieties bind in hydrated cavities, forming water-mediated hydrogen bonds to the protein. The design of compounds to replace these water molecules is a new strategymore » for the further optimization of C. parvum inhibitors for both antiparasitic and antibacterial applications.« less

  11. AN EVALUATION OF CRYPTOSPORIDIUM PARVUM GENOTYPING

    EPA Science Inventory

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

  12. Cryptosporidium meleagridis: Infectivity in Healthy Adult Volunteers

    PubMed Central

    Chappell, Cynthia L.; Okhuysen, Pablo C.; Langer-Curry, Rebecca C.; Akiyoshi, Donna E.; Widmer, Giovanni; Tzipori, Saul

    2011-01-01

    Most Cryptosporidium infections in humans are caused by C. parvum or C. hominis. However, genotyping techniques have identified infections caused by unusual Cryptosporidium species. Cryptosporidium meleagridis has been identified in ≤ 1% of persons with diarrhea, although prevalence is higher in developing nations. We examined the infectivity of C. meleagridis in healthy adults. Five volunteers were challenged with 105 C. meleagridis oocysts and monitored six weeks for fecal oocysts and clinical manifestations. Four volunteers had diarrhea; three had detectable fecal oocysts; and one infected volunteer remained asymptomatic. Fecal DNA from two volunteers was amplified by using a polymerase chain reaction specific for the Cryptosporidium small subunit ribosomal RNA gene. Nucleotide sequence of these amplicons was diagnostic for C. meleagridis. All infections were self-limited; oocysts were cleared within ≤ 12 days of challenge. These studies establish that healthy adults can be infected and become ill from ingestion of C. meleagridis oocysts. PMID:21813841

  13. DETECTION OF CRYPTOSPORIDIUM OOCYSTS IN WATER MATRICES

    EPA Science Inventory

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

  14. INACTIVATION OF CRYPTOSPORIDIUM PARVUM OOCYSTS WITH OZONE

    EPA Science Inventory

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

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

    PubMed Central

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

    2015-01-01

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

  16. Occurrence of Giardia and Cryptosporidium in water supplies of Russia and Bulgaria.

    PubMed

    Karanis, P; Sotiriadou, I; Kartashev, V; Kourenti, C; Tsvetkova, N; Stojanova, K

    2006-11-01

    The aim of the present study was to investigate water supplies in southern Russia and Bulgaria, in order to estimate the occurrence of Giardia and Cryptosporidium in drinking water resources from these countries. A total of 166 water samples of different origin (surface, tap, bottled, well, spring and waste water) were collected from Rostov (southern Russia), Sofia and Varna (Bulgaria) Greater Areas and screened for the detection of Giardia cysts and Cryptosporidium oocysts. The method incorporated concentration of water samples by filtration and flocculation, sucrose purification, (oo)cyst detection/identification by immunofluorescence test and differential interference contrast. Sixteen out of 166 samples (9.6%) were positive for Giardia and 30 (18.1%) positive for Cryptosporidium. Both Giardia and Cryptosporidium were detected in tap, river, well and waste waters. Giardia cysts were additionally detected in bottled water. Particularly some river, waste and well water samples were highly contaminated with (oo)cysts. This study has shown that drinking water supplies in Russia and Bulgaria are subject to contamination with Giardia and Cryptosporidium, with potential hazards for the public health. PMID:16780829

  17. Polymeric additives for antistatic treatment of sheet molding composites (SMC)

    NASA Astrophysics Data System (ADS)

    Gedan-Smolka, Michaela; Schubert, Katrin; Taeger, Antje; Marks, Hagen

    2016-03-01

    Novel oligomeric/polymeric additives with ionic units were synthesized in a one pot procedure in order to enhance the specific surface resistivity of SMC-materials. In contrast to industrially used inorganic salts or carbon materials, such as carbon black or CNT's, these substances are nearly colorless and matrix compatible and, resulting from that, easy to homogenize with the SMC-matrix. Furthermore, it was found, that additives which can be covalently bonded to the matrix during the molding procedure are preferred compared to physically mixed substances. The effectiveness of chemically bonded substances regarding with the lowering of the specific surface resistivity is higher and selected additives do not change the reactivity and resulting mechanical properties of the molded material.

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

    SciTech Connect

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

    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.

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

    EPA Science Inventory

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

  20. Development of a multilocus sequence typing tool for Cryptosporidium muris and Cryptosporidium andersoni

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Although widely used in the characterization of the transmission of intestinal Cryptosporidium spp., genotyping tools are not available for C. muris and C. andersoni, two most common gastric Cryptosporidium spp. of mammals. In this study, we screened the C. muris whole genome sequencing data for mi...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  2. [New methods for the diagnosis of Cryptosporidium and Giardia].

    PubMed

    Cacciò, S M

    2004-06-01

    The accurate identification of a parasite at the species and/or genotype level has major implications for various aspects of human and veterinary parasitology, including the diagnosis, the taxonomy, the treatment and the control. The advent of molecular techniques, in particular those based on the in vitro amplification of nucleic acids, has dramatically improved our ability to detect infections caused by parasites. To illustrate the progress in molecular diagnostics, Cryptosporidium and Giardia are used here as examples of parasites for which both the diagnosis and the taxonomy have traditionally been problematic. These protozoan parasites, while very different for many aspects of their biology, shares a complex series of transmission routes, including anthroponotic and zoonotic transmission, as well as waterborne and foodborne transmission. The resistant stages produced by Cryptosporidium and Giardia (oocysts and cysts, respectively) are remarkably stable, and can survive for weeks to months in the environment. Further, the infective dose is low, and infectious dose studies and models suggest that even a single oocyst or cyst carries some probability of causing an infection. Finally, most faeces that contain (oo)cysts end up in the environment and can be spread to foods by irrigation or by direct contact, and can persist in the water, as routine treatments eliminate only a fraction of these stages. This situation explains the growing interest towards the development of methods that allows such stages to be detected with the highest sensitivity and specificity. A variety of Polymerase Chain Reaction (PCR) assays have been described for both Cryptosporidium and Giardia. The choice of a particular assay mainly depends on the amount of information carried by the genetic locus under analysis. Indeed, some assays can be used to identify the different species within a genus, while others allowed to distinguish between isolates of the same species (genotypes), and some

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

    PubMed Central

    2010-01-01

    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. PMID:20338035

  4. Prevalence and Genetic Characterization of Cryptosporidium Isolates from Common Brushtail Possums (Trichosurus vulpecula) Adapted to Urban Settings▿

    PubMed Central

    Hill, Nichola J.; Deane, Elizabeth M.; Power, Michelle L.

    2008-01-01

    The common brushtail possum (Trichosurus vulpecula) is one of the most abundant native marsupials in urban Australia, having successfully adapted to utilize anthropogenic resources. The habituation of possums to food and shelter available in human settlements has facilitated interaction with people, pets, and zoo animals, increasing the potential for transmission of zoonotic Cryptosporidium pathogens. This study sought to examine the identity and prevalence of Cryptosporidium species occurring in possums adapted to urban settings compared to possums inhabiting remote woodlands far from urban areas and to characterize the health of the host in response to oocyst shedding. Findings indicated that both populations were shedding oocysts of the same genotype (brushtail possum 1 [BTP1]) that were genetically and morphologically distinct from zoonotic species and genotypes and most closely related to Cryptosporidium species from marsupials. The urban population was shedding an additional five Cryptosporidium isolates that were genetically distinct from BTP1 and formed a sister clade with Cryptosporidium parvum and Cryptosporidium hominis. Possums that were shedding oocysts showed no evidence of pathogenic changes, including elevated levels of white blood cells, diminished body condition (body mass divided by skeletal body length), or reduced nutritional state, suggesting a stable host-parasite relationship typical of Cryptosporidium species that are adapted to the host. Overall, Cryptosporidium occurred with a higher prevalence in possums from urban habitat (11.3%) than in possums from woodland habitat (5.6%); however, the host-specific nature of the genotypes may limit spillover infection in the urban setting. This study determined that the coexistence of possums with sympatric populations of humans, pets, and zoo animals in the urban Australian environment is unlikely to present a threat to public health safety. PMID:18641156

  5. PCR slippage across the ML-2 microsatellite of the Cryptosporidium MIC1 locus enables development of a PCR assay capable of distinguishing the zoonotic Cryptosporidium parvum from other human infectious Cryptosporidium species.

    PubMed

    Webber, M A; Sari, I; Hoefel, D; Monis, P T; King, B J

    2014-08-01

    Cryptosporidium are ubiquitous and significant enteropathogens of all classes of vertebrates and a major cause of human morbidity and mortality worldwide. Of the 24 recognized species, the zoonotic Cryptosporidium parvum and the host-specific Cryptosporidium hominis cause the majority of cases of human cryptosporidiosis. Here, we report on structural and transcriptional variability between C. parvum and C. hominis at the MIC1 locus, which encodes a microneme localized thrombospondin-like domain containing protein previously demonstrated to be critical for host cell infection by C. parvum. We demonstrate, using reverse transcription quantitative PCR with the aid of genomic data from the EuPathDB site, that the transcribed product in C. hominis is both truncated and significantly down-regulated in the sporozoite. We hypothesize that CpMIC1 may be a genetic factor involved in facilitating the wider host range of C. parvum in comparison with the specific host range of C. hominis. Furthermore, we show that the presence of a microsatellite (ML-2) within the C. parvum MIC-1 locus enables the development of a PCR marker that can rapidly distinguish the zoonotic C. parvum from C. hominis and other significant human infectious Cryptosporidium species due to reproducible PCR slippage across the ML-2 microsatellite. Additionally, we demonstrate that this locus is tightly linked to the GP60 locus, a locus commonly used in the genetic characterization of C. parvum and C. hominis isolates. This marker should provide a robust and additional tool to aid in the rapid identification of C. parvum from other Cryptosporidium species. PMID:23954136

  6. DEVELOPMENT OF A CT EQUATION FOR THE INACTIVATION OF CRYPTOSPORIDIUM OOCYSTS WITH OZONE

    EPA Science Inventory

    Cryptosporidium parvum, a protozoan parasite, has been implicated in a number of disease outbreaks. It is difficult to inactivate using free chlorine, but appears to be inactivated by ozone. Therefore, the USEPA has promulgated the Interim Enhanced Surface Water Treatment Rule, w...

  7. INACTIVATION OF CRYPTOSPORIDIUM PARVUM OOCYSTS WITH OZONE AND MONOCHLORAMINE AT LOW TEMPERATURE. (R826830)

    EPA Science Inventory

    The rate of Cryptosporidium parvum inactivation decreased with decreasing temperature (1¯20°C) for ozone and for monochloramine applied alone as well as after pre-treatment with ozone. Synergy was observed at all temperatures studied for the ozone/m...

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

    PubMed Central

    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

  9. GUIDANCE DOCUMENT ON IMPLEMENTATION OF THE MICROBIAL TOOLBOX FOR COMPLIANCE WITH LONG TERM 2 ENHANCED SURFACE WATER TREATMENT RULE (LT2ESWTR)

    EPA Science Inventory

    The Agreement in Principle for the Stage 2 M-DBP Federal Advisory Committee contains a list of treatment processes and management practices for water systems to use in meeting additional Cryptosporidium treatment requirements under the LT2ESWTR. This list, termed the microbial t...

  10. A HAPPY Map of Cryptosporidium parvum

    PubMed Central

    Piper, Michael B.; Bankier, Alan T.; Dear, Paul H.

    1998-01-01

    We have constructed a HAPPY map of the apicomplexan parasite Cryptosporidium parvum. We have placed 204 markers on the 10.4-Mb genome, giving an average marker spacing of ∼50 kb, with an effective resolution of ∼40 kb. HAPPY mapping (an in vitro linkage technique based on screening approximately haploid amounts of DNA by the polymerase chain reaction) is fast and accurate and is not subject to the distortions inherent in cloning, meiotic recombination, or hybrid cell formation. In addition, little genomic DNA is needed as a substrate, and the AT content of the genome is largely immaterial, making it an ideal method for mapping otherwise intractable parasite genomes. The map, covering all eight chromosomes, consists of 10 linkage groups, each of which has been chromosomally assigned. We have verified the accuracy of the map by several methods, including the construction of a >140-kb PAC contig on chromosome VI. Less than 1% of our markers detect non-rDNA duplicated sequences. PMID:9872984

  11. Investigation of Cryptosporidium spp. antigen by ELISA method in stool specimens obtained from patients with diarrhea.

    PubMed

    Elgun, Gullu; Koltas, Ismail Soner

    2011-02-01

    Cryptosporidium spp. is an important parasitic protozoan causing diarrhea in developing and developed countries. The agent causes severe life-threatening diarrhea especially in immunocompromised hosts. Diagnosis of the Cryptosporidium oocyst in stool samples by conventional microscopy is labor-intensive and time-consuming. Thus, we aimed to evaluate the usefulness of a copro-antigen enzyme-linked immunosorbent assay (ELISA) test in detecting Cryptosporidium spp. from fecal specimens. For this aim, microscopy and specific antigen detection methods were compared to determine Cryptosporidium spp. In addition, specific antigen by ELISA method in stool was investigated in order to find out whether or not it contributes to the diagnosis of Cryptosporidium spp. One hundred and fifty-four stool specimens taken from patients whose ages ranged from 0 to 86 with diarrhea applied to Department of Parasitology, Balcali Hospital of Cukurova University in Adana, Turkey were used. All samples were examined for Cryptosporidium spp. antigen by ELISA and oocysts via gold standard modified acid-fast staining, between October 2008 and July 2009. Eight (5.19%) specimens were found to be positive by modified acid-fast staining method and 37 (24.03%) specimens by copro-antigen ELISA method were found to be positive. The sensitivity and specificity for copro-antigen ELISA were 100% and 80.1%, respectively. The results of copro-antigen ELISA indicate that the simple, rapid, reliable, and standardized immunoassay test is sensitive and specific for routine diagnosis and may be useful for large-scale epidemiological studies of cryptosporidiosis. PMID:20938687

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

    PubMed Central

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

    2004-01-01

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

  13. Cryptosporidium, Giardia, and Cyclospora in ancient Peruvians.

    PubMed

    Ortega, Ynes R; Bonavia, Duccio

    2003-06-01

    Twenty-two coprolites of human origin, collected from excavations along the north-central coast of Peru, were examined using fluorescent microscopy for the presence of fecal parasites, with emphasis on Cryptosporidium sp., Giardia sp., and Cyclospora sp. Three samples were positive. One coprolite dated between ca. 2,375 and 1,525 BC contained Giardia sp. cysts. This coprolite corresponded to the Peruvian preceramic period. Another positive coprolite ca. AD 770-830 corresponded to Epoch 3 of the Middle Horizon and contained Cryptosporidium sp. oocysts. The third positive coprolite (corresponding to the Middle Horizon. ca. AD 500-900) contained Giardia sp. cysts. This report demonstrates that Giardia sp. and Cryptosporidium sp. were present in Peruvian coastal populations for at least 4,300 and 1,100 BP. PMID:12880276

  14. Molecular epidemiology of Giardia and Cryptosporidium infections.

    PubMed

    Thompson, R C A; Ash, A

    2016-06-01

    Giardia and Cryptosporidium are ubiquitous enteric protozoan pathogens of vertebrates. Although recognised as the aetiological agents of disease in humans and domestic animals for many years, fundamental questions concerning their ecology have been unresolved. Molecular tools have helped to better understand their genetic diversity and in so doing have helped to resolve questions about their transmission patterns and associated impacts on public health. However, the value of molecular tools is often complicated by questions concerning their applications, interpretation of results and terminology. Taxonomic issues have, until recently, made it difficult to determine the epidemiology of infections with both Giardia and Cryptosporidium. Similarly, improved understanding of their respective phylogenetic relationships has helped to resolve questions about zoonotic potential and distribution in wildlife. In the case of Cryptosporidium, imaging technologies have complemented phylogenetic studies in demonstrating the parasite's affinities with gregarine protozoa and have further supported its extracellular developmental capability and potential role as an environmental pathogen. PMID:26458528

  15. Environmental ecology of Cryptosporidium and public health implications.

    PubMed

    Rose, J B

    1997-01-01

    Cryptosporidium has become the most important contaminant found in drinking water and is associated with a high risk of waterborne disease particularly for the immunocompromised. There have been 12 documented waterborne outbreaks in North America since 1985; in two of these (Milwaukee and Las Vegas) mortality rates in the immunocompromised ranged from 52% to 68%. The immunofluorescence antibody assay (IFA) using epifluorescence microscopy has been used to examine the occurrence of Cryptosporidium in sewage (1 to 120 oocysts/liter), filtered secondary treated wastewater (0.01 to 0.13 oocysts/liter), surface waters (0.001 to 107 oocysts/liter), groundwater (0.004 to 0.922 oocysts/liter) and treated drinking water (0.001 to 0.72 oocysts/liter). New rules are being developed (Information Collection Rule and Enhanced Surface Water Treatment Rule) to obtain more occurrence data for drinking water systems for use with new risk assessment models. Public health officials should consider a communication program to physicians treating the immunocompromised, nursing homes, develop a plan to evaluate cases of cryptosporidiosis in the community, and contribute to the development of public policies that limit contamination of source waters, improve water treatment, and protect public health. PMID:9143715

  16. Cryptosporidium and Giardia as foodborne zoonoses.

    PubMed

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

    2007-10-21

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

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

    EPA Science Inventory

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

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

    PubMed Central

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

    2013-01-01

    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. PMID:23916060

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

    PubMed

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

    2013-11-01

    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. PMID:23916060

  20. Unexpected finding of feline-specific Giardia duodenalis assemblage F and Cryptosporidium felis in asymptomatic adult cattle in Northern Spain.

    PubMed

    Cardona, Guillermo A; de Lucio, Aida; Bailo, Begoña; Cano, Lourdes; de Fuentes, Isabel; Carmena, David

    2015-04-30

    Giardia duodenalis and Cryptosporidium spp. are common enteric protozoan parasites in production animals, including cattle. Typically, both the clinical outcome of these infections and the distribution of G. duodenalis assemblages and Cryptosporidium species are age-dependent, with the occurrence of diarrhoeal disease being mainly associated with young animals and sub-clinical, low intensity infections being predominantly found in adult animals. To investigate the prevalence and genetic diversity of G. duodenalis and Cryptosporidium spp. in asymptomatic adult cattle, a total of 362 faecal samples were collected in four farms in the province of Álava, Northern Spain, between November 2011 and December 2012. The presence of G. duodenalis was estimated by real-time PCR, and the assemblages were determined by multilocus sequence-based genotyping of the glutamate dehydrogenase and β-giardin genes of the parasite. Detection and identification of Cryptosporidium species was carried out by sequencing of a partial fragment of the small-subunit ribosomal RNA gene. Overall, G. duodenalis and Cryptosporidium spp. were detected in 68 (18.8%) and 45 (12.4%), respectively, of the 362 animals tested. Strikingly, four isolates representing two novel sub-types of G. duodenalis assemblage F were identified at the gdh, but not the bg, locus. This is the first report describing the presence of the feline-specific G. duodenalis assemblage F in bovine isolates. Additionally, five (three novel and two known) sub-types of G. duodenalis assemblage E were also identified at the gdh locus and a single one (assigned to sub-assemblage EII) at the bg locus. Of nine Cryptosporidium isolates, four (including a novel sub-type) were assigned to the cat-specific C. felis, two were typed as C. bovis, and the remaining three were only characterized at the genus level. Data presented here provide epidemiological and molecular evidence demonstrating that the host range specificity of G. duodenalis

  1. Direct and indirect QMRA of infectious Cryptosporidium oocysts in reclaimed water.

    PubMed

    Agulló-Barceló, M; Casas-Mangas, R; Lucena, F

    2012-12-01

    Water scarcity leads to an increased use of reclaimed water, which in turn calls for an improvement in water reclamation procedures to ensure adequate quality of the final effluent. The presence of infectious Cryptosporidium oocysts (IOO) in reclaimed water is a health hazard for users of this resource. Here, we gathered information on Cryptosporidium (concentrations, infectivity and genotype) in order to perform quantitative microbial risk assessment (QMRA). Moreover, data concerning the spores of sulphite-reducing clostridia (SRC) were used to undertake QMRA at a screening level. Our results show that the probability of infection (PI) by Cryptosporidium depends on the tertiary treatment type. The mean PI using the exponential dose-response model was 3.69 × 10(-6) in tertiary effluents (TE) treated with UV light, whereas it was 3 log(10) units higher, 1.89 × 10(-3), in TE not treated with this disinfection method. With the β-Poisson model, the mean PI was 1.56 × 10(-4) in UV-treated TE and 2 log(10) units higher, 4.37 × 10(-2), in TE not treated with UV. The use of SRC to perform QMRA of Cryptosporidium showed higher PI than when using directly IOO data. This observation suggests the former technique is a conservative method of QMRA. PMID:23165711

  2. A review of the global burden, novel diagnostics, therapeutics, and vaccine targets for cryptosporidium

    PubMed Central

    Checkley, William; White, A Clinton; Jaganath, Devan; Arrowood, Michael J; Chalmers, Rachel M; Chen, Xian-Ming; Fayer, Ronald; Griffiths, Jeffrey K; Guerrant, Richard L; Hedstrom, Lizbeth; Huston, Christopher D; Kotloff, Karen L; Kang, Gagandeep; Mead, Jan R; Miller, Mark; Petri, William A; Priest, Jeffrey W; Roos, David S; Striepen, Boris; Thompson, R C Andrew; Ward, Honorine D; Van Voorhis, Wesley A; Xiao, Lihua; Zhu, Guan; Houpt, Eric R

    2015-01-01

    Cryptosporidium spp are well recognised as causes of diarrhoeal disease during waterborne epidemics and in immunocompromised hosts. Studies have also drawn attention to an underestimated global burden and suggest major gaps in optimum diagnosis, treatment, and immunisation. Cryptosporidiosis is increasingly identified as an important cause of morbidity and mortality worldwide. Studies in low-resource settings and high-income countries have confirmed the importance of cryptosporidium as a cause of diarrhoea and childhood malnutrition. Diagnostic tests for cryptosporidium infection are suboptimum, necessitating specialised tests that are often insensitive. Antigen-detection and PCR improve sensitivity, and multiplexed antigen detection and molecular assays are underused. Therapy has some effect in healthy hosts and no proven efficacy in patients with AIDS. Use of cryptosporidium genomes has helped to identify promising therapeutic targets, and drugs are in development, but methods to assess the efficacy in vitro and in animals are not well standardised. Partial immunity after exposure suggests the potential for successful vaccines, and several are in development; however, surrogates of protection are not well defined. Improved methods for propagation and genetic manipulation of the organism would be significant advances. PMID:25278220

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

    PubMed

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

    2016-09-01

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

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

    NASA Astrophysics Data System (ADS)

    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.

  5. IDENTIFICATION OF 'CRYPTOSPORIDIUM' OOCYSTS IN RIVER WATER

    EPA Science Inventory

    Water samples were collected from four rivers in Washington State and two rivers in California and examined for the presence of Cryptosporidium oocysts. Oocyst-sized particles were concentrated from 20-liter samples of water by membrane filtration, centrifugation, and differentia...

  6. Molecular characterization of cryptosporidium in brazilian sheep

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Feces were collected from 125 sheep between January and December 2007, on ten farms in the State of Rio de Janeiro, Brazil, and examined for the presence of Cryptosporidium. Ninety samples were collected from lambs 2 to 6 months of age, and 35 were from sheep over 12 months of age. All samples were...

  7. Experimental infection of piglets with cryptosporidium.

    PubMed

    Tzipori, S; McCartney, E; Lawson, G H; Rowland, A C; Campbell, I

    1981-11-01

    Piglets from five litters were doses orally with cryptosporidium originally derived from diarrhoeic calves. The piglets were either nursed by the sow, artificially reared after sucking colostrum, or weaned on to creep feed. Colostrum-fed, artificially reared piglets obtained from two litters and exposed in the first week of life developed clinical signs of inappetence, vomiting and diarrhoea and shed oocysts in the faeces. Histologically the parasite was observed throughout the small and large intestine attached to epithelial cell surfaces and its presence was associated with extensive mucosal damage, particularly in the posterior small intestine, stunting and fusion of villi, immaturity of villous epithelial cells and oedema with increased cellularity of the lamina propria. Piglets from two other litters, both sucking and colostrum-fed artificially reared, exhibited similar but milder clinical signs. Histological lesions were less severe and cryptosporidium infection less extensive. When weaned piglets were exposed they remained clinically healthy although histologically there was evidence of cryptosporidium attachment in the small intestine and minor mucosal damage. There appears to be a good correlation between the extent of intestinal infection, the degree of mucosal damage and the severity of clinical disease induced by cryptosporidium in piglets. PMID:7342229

  8. Molecular Genotyping of Viable Cryptosporidium Oocysts

    EPA Science Inventory

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

  9. Occurrence of Cryptosporidium andersoni in Brazilian cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Feces were collected from 68 cattle, 1 to 12 mo of age, on 12 farms in the municipality of Campos dos Goytacazes, Rio de Janeiro, Brazil, and examined for the presence of Cryptosporidium sp. All samples were subjected to molecular analysis by polymerase chain reaction (nested PCR) of the 18S rRNA. F...

  10. Infectivity of the cervine genotype of Cryptosporidium

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  11. CRYPTOSPORIDIUM IN CATTLE FROM OBSERVING TO UNDERSTANDING

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  12. RELIABILITY OF SURROGATES FOR DETERMINING CRYPTOSPORIDIUM REMOVAL

    EPA Science Inventory

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

  13. Age related susceptibility of pigs to Cryptosporidium scrofarum infection.

    PubMed

    Kváč, Martin; Němejc, Karel; Kestřánová, Michaela; Květoňová, Dana; Wagnerová, Pavla; Kotková, Michaela; Rost, Michael; Samková, Eva; McEvoy, John; Sak, Bohumil

    2014-05-28

    Piglets from 4 to 8 weeks of age originated from a Cryptosporidium-free research breed were orally inoculated with 1 × 10(6) infectious oocysts of Cryptosporidium scrofarum. The number of shed oocysts per gram of faeces served to describe the infection intensity and prepatent period. In addition, faecal samples collected daily and tissue samples of the small and large intestine collected at 30 days post-inoculation were examined for the C. scrofarum small subunit ribosomal RNA gene using PCR. The piglets inoculated at 4-weeks of age remained uninfected, whereas 5-week-old and older animals were fully susceptible with a prepatent period ranging from 4 to 8 days. Susceptible pigs shed oocysts intermittently, and shedding intensity, reaching a mean maximum of 6000 oocysts per gram, did not differ significantly among infected animals. This study demonstrates that pigs become susceptible to C. scrofarum infection as late as 5-weeks of age. The mechanisms of age related susceptibility remain unknown. PMID:24630710

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

    PubMed

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

    2013-10-18

    Faecal samples from two horse farms in Algeria keeping Arabian, Thoroughbred, and Barb horses were examined for the presence of Cryptosporidium in 2010-2011. A total of 138 faecal samples (16 from a farm keeping 50 animals and 122 from a farm with 267 horses) were screened for Cryptosporidium spp. infection using molecular tools. DNA was extracted from all samples. Nested PCR was performed to amplify fragments of the SSU rDNA and gp60 genes to determine the presence of Cryptosporidium species and genotypes. Sequence analyses of SSU and gp60 genes revealed four animals positive for the presence of subtype XIIIa A22R9 of the Cryptosporidium hedgehog genotype. The infections were not associated with diarrhoea. This study reports, for the first time, the occurrence of Cryptosporidium in Algeria and the first occurrence of the hedgehog genotype in horses. These findings support the potential role of infected horses in sylvatic-domestic transmission of Cryptosporidium. PMID:23731858

  15. Detection and quantification of Cryptosporidium oocysts in environmental surfaces of an Equine Perinatology Unit.

    PubMed

    Piva, Silvia; Caffara, Monica; Pasquali, Frédérique; Castagnetti, Carolina; Iacono, Eleonora; Massella, Elisa; Zanoni, Renato Giulio; Galuppi, Roberta

    2016-09-01

    The presence of Cryptosporidium in institutions such as veterinary teaching hospitals, where students and staff are in frequent contact with animals, could represent a serious public health risk. In this study the detection and quantification of the Cryptosporidium oocysts present on the environmental surfaces of an Equine Perinatology Unit (EPU) were investigated. During 3 foaling seasons 175 samples obtained by swabbing an area of the floor and walls of boxes and utility rooms of EPU with sterile gauze, in 3 different moments. Samples were collected at the end of foaling season (July), after washing procedures (September) and after washing and disinfecting procedures, at the beginning of a new foaling season (December). All the samples were subjected to nested-PCR, followed by genotyping and sub-typing methods and to qPCR, allowing the oocyst quantification. Cryptosporidium spp. was detected in 14 samples, of which 11 were from walls and three were from floors. The highest number of oocysts was found in a sample collected from the floor of one utility room used for setting up therapies and treatments. In most cases, oocyst numbers, estimated by qPCR, were reduced or eliminated after washing and disinfecting procedures. The genotyping and sub-typing methods allowed identification of 2 subtypes of C. parvum (IIaA15G2R1 and IIdA23G1) and 1 of Cryptosporidium horse genotype (VIaA15G4) that were described in foals hospitalized at the EPU in the same years. The results of the present study show that qPCR can be used to evaluate Cryptosporidium contamination of environmental surfaces of a veterinary teaching hospital and the efficacy of the disinfection procedures. PMID:27544254

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

    PubMed Central

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

    2015-01-01

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

  17. Microsporidia and Cryptosporidium in horses and donkeys in Algeria: detection of a novel Cryptosporidium hominis subtype family (Ik) in a horse.

    PubMed

    Laatamna, Abd Elkarim; Wagnerová, Pavla; Sak, Bohumil; Květoňová, Dana; Xiao, Lihua; Rost, Michael; McEvoy, John; Saadi, Ahmed Rachid; Aissi, Meriem; Kváč, Martin

    2015-03-15

    A total of 219 and 124 individual fecal samples of horses and donkeys, respectively, were screened for the presence of Cryptosporidium spp., Encephalitozoon spp., and Enterocytozoon bieneusi DNA by genus-specific nested PCR. Isolates were genotyped by sequence analysis of SSU rRNA, GP60, TRAP-C1, COWP, and HSP70 loci in Cryptosporidium, and the ITS region in microsporidia. Cryptosporidium spp. was detected on 3/18 horse farms and 1/15 farms where donkeys were kept. Overall, five (2.3%) horse and two (1.6%) donkey specimens were PCR positive for Cryptosporidium. Genotyping at SSU and GP60 loci revealed that three isolates from horses and donkeys were C. parvum subtype family IIaA16G1R1, one isolate from a horse was, C. muris RN66, and one isolate from a donkey was C. muris TS03. An isolate from a horse shared 99.4% and 99.3% similarity with Cryptosporidium hominis and C. cuniculus, respectively, at the SSU locus. This isolate shared 100% identity with C. hominis at the TRAP-C1, COWP, and HSP70 loci, and it was from the novel gp60 subtype family IkA15G1. Microsporidia were found on 6/18 horse and 2/15 donkey farms. E. bieneusi was identified in 6.8% (15/219) and 1.6% (2/124), and Encephalitozoon cuniculi was identified in 1.8% (4/219) and 1.6% (2/124), of horses and donkeys, respectively. Three genotypes of E. cuniculi (I, II and III) were detected in horses, and E. cuniculi genotype II was detected in donkeys. Four genotypes of E. bieneusi (horse1, horse 2, CZ3, D) were described in horses. An additional five horses and two donkeys were positive for E. bieneusi, but the isolated were not genotyped. Neither Cryptosporidium nor microsporidia prevalence were affected by sex, age, type of breeding, or whether the host was a horse or a donkey. PMID:25638716

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

    PubMed

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

    2015-01-01

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

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

    PubMed Central

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

    2015-01-01

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

  20. Cryptosporidium rubeyi n. sp. (Apicomplexa: Cryptosporidiidae) in multiple Spermophilus ground squirrel species.

    PubMed

    Li, Xunde; Pereira, Maria das Graças Cabral; Larsen, Royce; Xiao, Chengling; Phillips, Ralph; Striby, Karl; McCowan, Brenda; Atwill, Edward R

    2015-12-01

    Previously we reported the unique Cryptosporidium sp. "c" genotype (e.g., Sbey03c, Sbey05c, Sbld05c, Sltl05c) from three species of Spermophilus ground squirrel (Spermophilus beecheyi, Spermophilus beldingi, Spermophilus lateralis) located throughout California, USA. This follow-up work characterizes the morphology and animal infectivity of this novel genotype as the final step in proposing it as a new species of Cryptosporidium. Analysis of sequences of 18S rRNA, actin, and HSP70 genes of additional Cryptosporidium isolates from recently sampled California ground squirrels (S. beecheyi) confirms the presence of the unique Sbey-c genotype in S. beecheyi. Phylogenetic and BLAST analysis indicates that the c-genotype in Spermophilus ground squirrels is distinct from Cryptosporidium species/genotypes from other host species currently available in GenBank. We propose to name this c-genotype found in Spermophilus ground squirrels as Cryptosporidium rubeyi n. sp. The mean size of C. rubeyi n. sp. oocysts is 4.67 (4.4-5.0) μm × 4.34 (4.0-5.0) μm, with a length/width index of 1.08 (n = 220). Oocysts of C. rubeyi n. sp. are not infectious to neonatal BALB/c mice and Holstein calves. GenBank accession numbers for C. rubeyi n. sp. are DQ295012, AY462233, and KM010224 for the 18S rRNA gene, KM010227 for the actin gene, and KM010229 for the HSP70 gene. PMID:26543805

  1. Cryptosporidium rubeyi n. sp. (Apicomplexa: Cryptosporidiidae) in multiple Spermophilus ground squirrel species

    PubMed Central

    Li, Xunde; Pereira, Maria das Graças Cabral; Larsen, Royce; Xiao, Chengling; Phillips, Ralph; Striby, Karl; McCowan, Brenda; Atwill, Edward R.

    2015-01-01

    Previously we reported the unique Cryptosporidium sp. “c” genotype (e.g., Sbey03c, Sbey05c, Sbld05c, Sltl05c) from three species of Spermophilus ground squirrel (Spermophilus beecheyi, Spermophilus beldingi, Spermophilus lateralis) located throughout California, USA. This follow-up work characterizes the morphology and animal infectivity of this novel genotype as the final step in proposing it as a new species of Cryptosporidium. Analysis of sequences of 18S rRNA, actin, and HSP70 genes of additional Cryptosporidium isolates from recently sampled California ground squirrels (S. beecheyi) confirms the presence of the unique Sbey-c genotype in S. beecheyi. Phylogenetic and BLAST analysis indicates that the c-genotype in Spermophilus ground squirrels is distinct from Cryptosporidium species/genotypes from other host species currently available in GenBank. We propose to name this c-genotype found in Spermophilus ground squirrels as Cryptosporidium rubeyi n. sp. The mean size of C. rubeyi n. sp. oocysts is 4.67 (4.4–5.0) μm × 4.34 (4.0–5.0) μm, with a length/width index of 1.08 (n = 220). Oocysts of C. rubeyi n. sp. are not infectious to neonatal BALB/c mice and Holstein calves. GenBank accession numbers for C. rubeyi n. sp. are DQ295012, AY462233, and KM010224 for the 18S rRNA gene, KM010227 for the actin gene, and KM010229 for the HSP70 gene. PMID:26543805

  2. Prevalence of Cryptosporidium oocysts and Giardia cysts in raw and treated sewage sludges.

    PubMed

    Amorós, Inmaculada; Moreno, Yolanda; Reyes, Mariela; Moreno-Mesonero, Laura; Alonso, Jose L

    2016-11-01

    Treated sludge from wastewater treatment plants (WWTPs) is commonly used in agriculture as fertilizers and to amend soils. The most significant health hazard for sewage sludge relates to the wide range of pathogenic microorganisms such as protozoa parasites.The objective of this study was to collect quantitative data on Cryptosporidium oocysts and Giardia cysts in the treated sludge in wastewater treatment facilities in Spain. Sludge from five WWTPs with different stabilization processes has been analysed for the presence of Cryptosporidium and Giardia in the raw sludge and after the sludge treatment. A composting plant (CP) has also been assessed. After a sedimentation step, sludge samples were processed and (oo)cysts were isolated by immunomagnetic separation (IMS) and detected by immunofluorescence assay (IFA). Results obtained in this study showed that Cryptosporidium oocysts and Giardia cysts were present in 26 of the 30 samples (86.6%) of raw sludge samples. In treated sludge samples, (oo)cysts have been observed in all WWTP's analysed (25 samples) with different stabilization treatment (83.3%). Only in samples from the CP no (oo)cysts were detected. This study provides evidence that (oo)cysts are present in sewage sludge-end products from wastewater treatment processes with the negative consequences for public health. PMID:27080207

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

    PubMed

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

    2016-01-30

    Cryptosporidium is a protozoan that can cause gastro-intestinal illness with diarrhoea in a wide range of hosts. In fact some species of Cryptosporidium can infect the broad range of hosts. The current paper is focused to investigate monthly prevalence and diversity of Cryptosporidium spp. during the spring and early summer (March-June) in 2009 and 2010 in farms with no history of cryptosporidiosis. Animal samples were analyzed to elucidate the prevalence of Cryptosporidium in two regions, West and the East catchments in Ireland. Our investigation demonstrates the prevalence ranges from 14% to 26% an early summer peak (June) was observed. Based on the findings of this study Cryptosporidium ryanae (in cattle, horses), and Cryptosporidium bovis/xiaoi followed by Cryptosporidium parvum (in sheep) were found to be the predominant species in asymptomatic cases. The circulation of other Cryptosporidium species such as C. parvum, C. bovis, C. ubiquitum, C. andersoni and Cryptosporidium horse and pig genotypes in livestock was investigated. PMID:26801590

  4. Cryptosporidium parvum and Cryptosporidium andersoni infection in naturally infected cattle of northwest Iran

    PubMed Central

    Mirzai, Yousef; Yakhchali, Mohammad; Mardani, Karim

    2014-01-01

    The protozoan intestinal parasite Cryptosporidium commonly infects cattle throughout the world and Iran. The present study was undertaken to determine the abundance and associated risk factors of Cryptosporidium infection in cattle herds of northwestern Iran. A total number of 246 fecal samples from 138 (56.1%) diarrheic (D) and 108 (43.9%) non-diarrheic (ND) cattle were randomly collected and examined by fecal smears stained with Ziehl-Neelsen. For molecular specification, DNA was extracted from collected Cryptosporidium oocysts and a fragment of 1325 bp in size from 18S rRNA gene was amplified. The overall prevalence of Cryptosporidium infection was 22.3% (55/246). The prevalence of Cryptosporidium infection in examined calves less than 6 month-old was significantly higher than adult cattle. C. parvum and C. andersoni were identified in 20.3% (50/246) and 2.03% (5/246) of examined cattle, respectively. The highest prevalence of C. parvum infection was found in D calves < 6 month-old (13.4%, 33/246), while C. andersoni was only detected in ND cattle (8.9%, 22/246). There was significant difference in the prevalence between male than female cattle. There was no significant difference between prevalence and seasons of investigation. It was concluded that C. parvum was the prevalent species in younger animals compared to older ones as a potentially zoonotic agent in the region. PMID:25568693

  5. Impact of Age, Gender, and Addition of Probiotics on Treatment Success for Helicobacter pylori in Children

    PubMed Central

    Weiner, Noam; Shaoul, Ron

    2015-01-01

    The primary objective of this study was to evaluate the effect of age, gender, and the use of probiotics with standard treatment regimen on Helicobacter pylori eradication. Based on endoscopic findings and clinical presentation, selected patients were treated with standard triple therapy (omeprazole, clarithromycin, and amoxicillin). Those who failed were offered a repeat treatment with omeprazole, metronidazole, and amoxicillin. After the publications of the possible advantages of probiotic treatment on H pylori eradication, the probiotic agent “Probiotica Forte” was routinely added to the treatment. Eradication was noted for 94/130 patients (72%) and for 128/197 patients (65%) with or without probiotic agent, respectively (P = .23). For second-line treatment eradication was noted in 33/46 (72%) and in 9/20 (45%) with or without probiotic agent, respectively (P = .053). The addition of probiotics may improve eradication success especially in addition to second-line treatment. PMID:27335981

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

    PubMed Central

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

    2014-01-01

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

  7. THE WIDE GEOGRAPHIC RANGE OF CRYPTOSPORIDIUM BOVIS AND THE DEER-LIKE GENOTYPE IN BOVINES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recent studies in the United States reported that of preweaned dairy calves infected with Cryptosporidium ~85% were infected with zoonotic C. parvum whereas of postweaned calves infected with Cryptosporidium, only 1% were infected with C. parvum. Cryptosporidium bovis (syn. Cryptosporidium bovine ...

  8. Cryptosporidium parvum: From foal to veterinary students.

    PubMed

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

    2016-03-30

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

  9. Methods for Detection and Control of Cryptosporidium in the Environment

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryptosporidiosis is a gastrointestinal disease of humans and many animals caused by protozoan parasites in the genus Cryptosporidium. Cryptosporidium has become the most important newly recognized pathogen in drinking water, detected in over 90% of the surface waters tested in the United States an...

  10. Cryptosporidium xiaoi n. sp. (Apicomplexa: Cryptosporidiidae) in sheep (Ovis aries)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A new species, Cryptosporidium xiaoi, is described from sheep. Oocysts of C. xiaoi, previously identified as the Cryptosporidium bovis-like genotype and as the ovine genotype from sheep in Australia and the United States are recorded as such in GenBank (AY587166, EU203216, DQ182597, AY741309, and DQ...

  11. Clinical and subclinical infections with Cryptosporidium in animals

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  12. EVALUATING CRYPTOSPORIDIUM AND GIARDIA CONCENTRATIONS IN COMBINED SEWER OVERFLOW

    EPA Science Inventory

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

  13. Detection of viable Cryptosporidium parvum oocysts by PCR.

    PubMed Central

    Wagner-Wiening, C; Kimmig, P

    1995-01-01

    PCR was used to detect and specifically identify a gene fragment from Cryptosporidium parvum. An 873-bp region of a 2,359-bp DNA fragment encoding a repetitive oocyst protein of C. parvum was shown to be specifically amplified in C. parvum. An excystation protocol before DNA extraction allowed the differentiation between live and dead Cryptosporidium parvum oocysts. PMID:8534121

  14. IMMUNOFLUORESCENCE DETECTION OF 'CRYPTOSPORIDIUM' OOCYSTS IN FECAL SMEARS

    EPA Science Inventory

    An indirect fluorescent antibody (IFA) procedure was developed for the detection of Cryptosporidium sp. oocysts in human, nonhuman primate, and bovine fecal smears. The procedure, which takes about 90 min to perform, involves the use of a rabbit antiserum against Cryptosporidium ...

  15. Quantification of Treatment Effect Modification on Both an Additive and Multiplicative Scale

    PubMed Central

    Girerd, Nicolas; Rabilloud, Muriel; Pibarot, Philippe; Mathieu, Patrick; Roy, Pascal

    2016-01-01

    Background In both observational and randomized studies, associations with overall survival are by and large assessed on a multiplicative scale using the Cox model. However, clinicians and clinical researchers have an ardent interest in assessing absolute benefit associated with treatments. In older patients, some studies have reported lower relative treatment effect, which might translate into similar or even greater absolute treatment effect given their high baseline hazard for clinical events. Methods The effect of treatment and the effect modification of treatment were respectively assessed using a multiplicative and an additive hazard model in an analysis adjusted for propensity score in the context of coronary surgery. Results The multiplicative model yielded a lower relative hazard reduction with bilateral internal thoracic artery grafting in older patients (Hazard ratio for interaction/year = 1.03, 95%CI: 1.00 to 1.06, p = 0.05) whereas the additive model reported a similar absolute hazard reduction with increasing age (Delta for interaction/year = 0.10, 95%CI: -0.27 to 0.46, p = 0.61). The number needed to treat derived from the propensity score-adjusted multiplicative model was remarkably similar at the end of the follow-up in patients aged < = 60 and in patients >70. Conclusions The present example demonstrates that a lower treatment effect in older patients on a relative scale can conversely translate into a similar treatment effect on an additive scale due to large baseline hazard differences. Importantly, absolute risk reduction, either crude or adjusted, can be calculated from multiplicative survival models. We advocate for a wider use of the absolute scale, especially using additive hazard models, to assess treatment effect and treatment effect modification. PMID:27045168

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

    PubMed

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

    2016-05-01

    Proventriculus and intestinal samples from 70 North American red-winged blackbirds (Agelaius phoeniceus; order Passeriformes) were examined for the presence of Cryptosporidium by PCR amplification and sequence analysis of the 18S ribosomal RNA (18S rRNA), actin, and 70-kDa heat shock protein (HSP70) genes. Twelve birds (17.1 %) were positive for the Cryptosporidium 18S rRNA gene: six birds were positive at the proventriculus site only and six birds were positive at the proventriculus and intestinal sites. Sequence analysis of the 18S rRNA, actin and HSP70 genes showed the presence of the gastric species Cryptosporidium galli in a single proventriculus sample and a closely related genotype, which we have named Cryptosporidium avian genotype VI, in all other positive samples. These findings contribute to our understanding of Cryptosporidium diversification in passerines, the largest avian order. PMID:26818945

  17. Addition of alkali to the hydrothermal-mechanochemical treatment of Eucalyptus enhances its enzymatic saccharification.

    PubMed

    Ishiguro, Maki; Endo, Takashi

    2014-02-01

    The effects of alkali on hydrothermal-mechanochemical treatment (hydrothermal treatment combined with wet-milling) were examined with the aim of improving pretreatment of lignocellulosic biomass before enzymatic saccharification. After enzymatic saccharification, the highest glucose yield was obtained by autoclaving at 170°C in the presence of 20% NaOH per substrate weight. The wood fiber was unraveled into finer nanofibers by hydrothermal-mechanochemical treatment, thus increasing the specific surface area of the substrate from 11 to 132m(2)/g. Adding 20% NaOH to the treatment further increased the specific surface area of the already fibrillated substrate by 76% (232m(2)/g) due to lignin removal and ester bond cleavage between lignin and hemicellulose. This increase in specific surface area was closely related to the increase in enzymatic digestibility; therefore, NaOH addition may have enhanced the effect of hydrothermal-mechanochemical treatment. PMID:24378778

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

    PubMed Central

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

    2009-01-01

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

  19. Detection of Cryptosporidium parvum and Cryptosporidium hominis in human patients in Cairo, Egypt.

    PubMed

    Abd El Kader, Nour M; Blanco, María-Alejandra; Ali-Tammam, Marwa; Abd El Ghaffar, Abd El Rahman B; Osman, Ahmed; El Sheikh, Nabila; Rubio, José Miguel; de Fuentes, Isabel

    2012-01-01

    Cryptosporidium is a significant cause of diarrheal disease in developing and industrialized nations. Cryptosporidium hominis and Cryptosporidium parvum are the main agents of cryptosporidiosis in humans. In Egypt, very little is known about genetic structure of Cryptosporidium spp. Therefore, this study was designed to examine samples from sporadic cases of cryptosporidiosis in Egyptians in order to identify the species involved in infection as well as the transmission dynamics and distribution of the parasite in the Great Cairo area. A total of 391 human faecal samples were collected, between May 2008 and March 2009, from ten public hospitals in Great Cairo. Initial screening by immunochromatographic detection kit "the Stick Crypto-Giardia; Operon" showed 23 possible positive cases. Twenty of them were confirmed by microscopic examination. PCR was performed by amplification of the oocyst wall protein (COWP) gene where 18 out of 23 samples were positive, one not detected by microscopy. Cryptosporidium genotyping was performed by RFLP analysis of PCR products of the diagnosis PCR. Only 15 samples rendered a digestion pattern. The genotyping distribution was nine cases showing C. hominis genotype, three showing C. parvum genotype and three showing mixed infection by C. hominis and C. parvum. The data showed an elevated prevalence of C. hominis (80.0%), the most anthroponotic species, suggesting a human-human transmission. Furthermore, the presence of up to 40% of samples infected with C. parvum shows that further investigations are required to determine the subgenotypes of C. parvum to clarify the mode of transmission in order to improve the control measures. PMID:21607688

  20. Pre-treatment factors associated with detecting additional brain metastases at stereotactic radiosurgery.

    PubMed

    Wardak, Zabi; Augustyn, Alexander; Zhu, Hong; Mickey, Bruce E; Whitworth, Louis A; Madden, Christopher J; Barnett, Samuel L; Abdulrahman, Ramzi E; Nedzi, Lucien A; Timmerman, Robert D; Choe, Kevin S

    2016-06-01

    The number of brain metastases identified on diagnostic magnetic resonance imaging (MRI) is a key factor in consideration of stereotactic radiosurgery (SRS). However, additional lesions are often detected on high-resolution SRS-planning MRI. We investigated pre-treatment clinical characteristics that are associated with finding additional metastases at SRS. Patients treated with SRS for brain metastases between the years of 2009-2014 comprised the study cohort. All patients underwent frame-fixed, 1 mm thick MRI on the day of SRS. Patient, tumor, and treatment characteristics were analyzed for an association with increase in number of metastases identified on SRS-planning MRI. 289 consecutive SRS cases were analyzed. 725 metastases were identified on pre-treatment MRI and 1062 metastases were identified on SRS-planning MRI. An increase in the number of metastases occurred in 34 % of the cases. On univariate analysis, more than four metastases and the diameter of the largest lesion were significantly associated with an increase in number of metastases on SRS-planning MRI. When stratified by the diameter of the largest lesion into <2, 2-3, or ≥3 cm, additional metastases were identified in 37, 29, and 18 %, respectively. While this increase in the number of metastases is largely due to the difference in imaging technique, the number and size of the metastases were also associated with finding additional lesions. These clinical factors may be considered when determining treatment options for brain metastases. PMID:26966096

  1. Sintering and properties of Si3N4 with and without additives by HIP treatment

    NASA Technical Reports Server (NTRS)

    Kuratani, S.; Shimada, M.; Koizumi, M.

    1986-01-01

    Hot Isostatic Pressing (HIP) of Si3N4 powders with and without additives was performed using a glass container, and various kinds of pressureless-sintered Si3N4 were HIP'ed without a container. The effects of HIP treatment on density, microstructure, flexural strength, microhardness, and fracture toughness on Si3N4 ceramics were studied. Using a glass container it was difficult to reach theoretical density. The microhardness of HIP'ed Si3N4 without additives was low, and the fracture toughness of HIP'ed Si3N4 with and without additives was 22 to 25 W/m-K, and it decreased with increasing the amount of additives. The density and flexural strength, and hardness of pressureless-sintered Si3N4 which contained Al2O and Y2O3 as oxide additives were remarkably improved by HIP treatment using nitrogen as a pressure transmitting gas. It is very important to select the sintering conditions for fabricating the presintered body of Si3N4 in order to improve the mechanical properties of Si3N4 by HIP treatment.

  2. Biology of Cryptosporidium parvum in pigs: from weaning to market.

    PubMed

    Guselle, N J; Appelbee, A J; Olson, M E

    2003-04-01

    Cryptosporidium parvum is commonly identified as infecting domestic livestock and humans. Prevalence of C. parvum in pigs has been reported, however, the duration and infection pattern of naturally acquired Cryptosporidium infections in pigs has not been reported. This study was undertaken to investigate the age of oocyst shedding and duration of natural Cryptosporidium parvum infections in pigs from weaning to market weight. Fecal samples were collected from weaned Yorkshire-Landrace piglets (n=33) twice per week until Cryptosporidium oocysts were detected. Upon oocyst detection, fecal samples were collected three times per week and pigs were monitored throughout the study for diarrhea and examined after concentration and immunofluroescent staining. Cryptosporidium isolates were genotyped by polymerase chain reaction to amplify the HSP70 gene which was subsequently sequence analyzed. All 33 pigs shed oocysts some time during the study. The mean age of initial oocyst detection was 45.2 days post-weaning with the mean duration of infection 28.7 days. Mean number of Cryptosporidium oocysts was low and declined to zero prior to study completion. Episodes of diarrhea were not associated with oocyst excretion. Genetic sequences were obtained for 10 of the pigs. All of the 10 isolates aligned as the Cryptosporidium parvum 'pig' genotype. This study demonstrates that the age and duration of oocyst shedding in pigs infected with C. parvum porcine genotype is different from other livestock species. PMID:12651214

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

    PubMed Central

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

    2001-01-01

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

  4. Development of a nested-PCR assay for the detection of Cryptosporidium parvum in finished water.

    PubMed

    Monis, P T; Saint, C P

    2001-05-01

    A nested-PCR assay, incorporating an internal positive control, was developed for Cryptosporidium monitoring in finished water. This assay was capable of reproducibly detecting 8 oocysts in spiked-filtered water samples collected from 5 South Australian water treatment plants. The RT-PCR assay of Kaucner and Stinear (Appl. Environ. Microbiol. 64(5) (1998) 1743) was also evaluated for the detection of Cryptosporidium parvum. Initially, under our experimental conditions, a detection level of 27 oocysts was achieved for spiked reagent water samples. This level was improved to 5 oocysts by modification of the method. Untreated South Australian source waters concentrated by calcium carbonate flocculation were found to be highly inhibitory to the RT-PCR assay. Concentration of similar samples using Envirochek filters appeared to eliminate PCR inhibition. While both methods possessed similar sensitivities the nested-PCR assay was more reproducible, more cost effective, simpler to perform and could detect both viable and non-viable intact Cryptosporidium parvum oocysts, which is an important consideration for plant operators. These factors make the nested-PCR assay the method of choice for screening large numbers of potable water samples, where a reliable low level of detection is essential. PMID:11329665

  5. Detection and molecular characterisation of Cryptosporidium parvum in British European hedgehogs (Erinaceus europaeus).

    PubMed

    Sangster, Lucy; Blake, Damer P; Robinson, Guy; Hopkins, Timothy C; Sa, Ricardo C C; Cunningham, Andrew A; Chalmers, Rachel M; Lawson, Becki

    2016-02-15

    Surveillance was conducted for the occurrence of protozoan parasites of the genus Cryptosporidium in European hedgehogs (Erinaceus europaeus) in Great Britain. In total, 108 voided faecal samples were collected from hedgehogs newly admitted to eight wildlife casualty treatment and rehabilitation centres. Terminal large intestinal (LI) contents from three hedgehog carcasses were also analysed. Information on host and location variables, including faecal appearance, body weight, and apparent health status, was compiled. Polymerase Chain Reaction (PCR) targeting the 18S ribosomal RNA gene, confirmed by sequencing, revealed an 8% (9/111) occurrence of Cryptosporidium parvum in faeces or LI contents, with no significant association between the host or location variables and infection. Archived small intestinal (SI) tissue from a hedgehog with histological evidence of cryptosporidiosis was also positive for C. parvum by PCR and sequence analysis of the 18S rRNA gene. No other Cryptosporidium species were detected. PCR and sequencing of the glycoprotein 60 gene identified three known zoonotic C. parvum subtypes not previously found in hedgehogs: IIdA17G1 (n=4), IIdA19G1 (n=1) and IIdA24G1 (n=1). These subtypes are also known to infect livestock. Another faecal sample contained C. parvum IIcA5G3j which has been found previously in hedgehogs, and for which there is one published report in a human, but is not known to affect livestock. The presence of zoonotic subtypes of C. parvum in British hedgehogs highlights a potential public health concern. Further research is needed to better understand the epidemiology and potential impacts of Cryptosporidium infection in hedgehogs. PMID:26827859

  6. Cryptosporidium and cryptosporidiosis: the African perspective.

    PubMed

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

    2016-07-01

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

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

    PubMed

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

    1990-04-01

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

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

    PubMed

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

    2016-01-01

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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

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

    2016-01-01

    Water monitoring is essential to ensure safe drinking water for consumers. However existing methods have several drawbacks, particularly with regard to the poor recovery of Cryptosporidium due to the inability to efficiently elute Cryptosporidium oocysts during the established detection process used by water utilities. Thus the development of new inexpensive materials that could be incorporated into the concentration and release stage that would control Cryptosporidium oocysts adhesion would be beneficial. Here we describe improved filter performance following dip-coating of the filters with a "bioactive" polyacrylate. Specifically 69% more oocysts were eluted from the filter which had been coated with a polymer than on the naked filter alone. PMID:26009471

  11. Unusual Enterocytozoon bieneusi genotypes and Cryptosporidium hominis subtypes in HIV-infected patients on highly active antiretroviral therapy.

    PubMed

    Akinbo, Frederick O; Okaka, Christopher E; Omoregie, Richard; Adamu, Haileeyesus; Xiao, Lihua

    2013-07-01

    Human immunodeficiency virus (HIV)-infected persons are commonly infected with Cryptosporidium species and Enterocytozoon bieneusi in both developed and developing countries, particularly patients with CD4+ cell counts below 200 cells/μL; 285 HIV-infected patients on highly active antiretroviral therapy (HAART) were enrolled in this study, and both stool and blood specimens were collected from participants. The stool specimens were analyzed and typed for E. bieneusi and Cryptosporidium spp. by polymerase chain reaction (PCR) and DNA sequencing. CD4 count was analyzed using flow cytometry. E. bieneusi and Cryptosporidium were detected in 18 (6.3%) and 4 (1.4%) patients, respectively. The E. bieneusi detected mostly belonged to a new genotype group that, thus far, has only been found in a few humans: genotype Nig4 in 2 patients and two new genotypes related to Nig4 in 12 patients. The Cryptosporidium detected included C. hominis (two patients), C. parvum (one patient), and C. felis (one patient), with the two C. hominis infections belonging to an unusual subtype family. Additional studies are required to determine whether some E. bieneusi genotypes and C. hominis subtypes are more prevalent in HIV patients on HAART. PMID:23629938

  12. Structure of Cryptosporidium IMP dehydrogenase bound to an inhibitor with in vivo antiparasitic activity

    SciTech Connect

    Kim, Youngchang; Makowska-Grzyska, Magdalena; Gorla, Suresh Kumar; Gollapalli, Deviprasad R.; Cuny, Gregory D.; Joachimiak, Andrzej; Hedstrom, Lizbeth

    2015-04-21

    Inosine 5´-monophosphate dehydrogenase (IMPDH) is a promising target for the treatment of Cryptosporidium infections. Here, the structure of C. parvum IMPDH (CpIMPDH) in complex with inosine 5´-monophosphate (IMP) and P131, an inhibitor with in vivo anticryptosporidial activity, is reported. P131 contains two aromatic groups, one of which interacts with the hypoxanthine ring of IMP, while the second interacts with the aromatic ring of a tyrosine in the adjacent subunit. In addition, the amine and NO2 moieties bind in hydrated cavities, forming water-mediated hydrogen bonds to the protein. The design of compounds to replace these water molecules is a new strategy for the further optimization of C. parvum inhibitors for both antiparasitic and antibacterial applications.

  13. CRYPTOSPORIDIUM LOG INACTIVATION CALCULATION METHODS

    EPA Science Inventory

    Appendix O of the Surface Water Treatment Rule (SWTR) Guidance Manual introduces the CeffT10 (i.e., reaction zone outlet C value and T10 time) method for calculating ozone CT value and Giardia and virus log inactivation. The LT2ESWTR Pre-proposal Draft Regulatory Language for St...

  14. Improved superconducting properties of melt-textured Nd123 by additional heat treatment

    NASA Astrophysics Data System (ADS)

    Chikumoto, N.; Yoshioka, J.; Murakami, M.

    1997-02-01

    We have investigated the effect of additional heat-treatment on the superconducting transition and the flux pinning properties of NdBaCuO melt-textured in air. After the heat-treatment at high temperatures, >900°C, under low oxygen partial pressure, P(O 2) = 0.001 atm, the superconducting transition became sharper accompanied by an increase of Jc. However, the increase of Jc was very small and the secondary peak effect commonly observed in NdBaCuO melt textured in low P(O 2) could not be observed. Transmission electron microscopic observations and energy dispersive X-ray analyses show that the spatial variation of the Nd/Ba ratio is reduced after high-temperature heat-treatment, which indicates that an improvement in Tc and Jc is attributed to a suppression of Nd substitution on the Ba site.

  15. Macitentan: An important addition to the treatment of pulmonary arterial hypertension

    PubMed Central

    Khadka, Anjan; Singh Brashier, Dick B; Tejus, Anantharamu; Sharma, Ashok Kumar

    2015-01-01

    Macitentan is an orphan drug for the treatment of pulmonary arterial hypertension (PAH). Endothelin-1 (ET-1) plays a critical role of pathophysiology of PAH. Macitentan, a new dual endothelin receptor antagonist, has reportedly improved prognosis of PAH patients by delaying the progression of disease. It prevents the binding of ET-1 to both endothelin A (ETA) and endothelin B (ETB) receptors. Macitentan displays higher efficacy, lesser adverse effects and drug interactions. It has completed phase III trials in 2012 for treatment of PAH and has been tried for ischemic digital ulcers in systemic sclerosis, recurrent glioblastoma and combination with chemotherapeutic agents against various cancers. Safety data for macitentan were obtained primarily from a placebo-controlled clinical study in 742 patients with PAH. The Food and Drug Administration (FDA) approved the drug on 13 October 2013. It is an important addition to long-term treatment of PAH. PMID:25709357

  16. Molecular genotyping and sub-genotyping of Cryptosporidium spp. isolates from symptomatic individuals attending two major public hospitals in Madrid, Spain.

    PubMed

    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

  17. ANIMAL SOURCE IDENTIFICATION USING A CRYPTOSPORIDIUM DNA CHARACTERIZATION TECHNIQUE

    EPA Science Inventory

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

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

    PubMed

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

    2013-01-01

    The aim of the present study was to diagnose the presence of Giardia cysts and Cryptosporidium oocysts in household animals using nested polymerase chain reaction (PCR) and sequence analysis. One hundred faecal samples obtained from 81 dogs and 19 cats were investigated. The Cryptosporidium genotypes were determined by sequencing a fragment of the small subunit (SSU) rRNA gene, while the Giardia Assemblages were determined through analysis of the glutamate dehydrogenase (GDH) locus. Isolates from five dogs and two cats were positive by PCR for the presence of Giardia, and their sequences matched the zoonotic Assemblage A of Giardia. Cryptosporidium spp. isolated from one dog and one cat were both found to be C. parvum. One dog isolate harboured a mixed infection of C. parvum and Giardia Assemblage A. These findings support the growing evidence that household animals are potential reservoirs of the zoonotic pathogens Giardia spp. and Cryptosporidium spp. for infections in humans. PMID:23477297

  19. Identification of a Novel Cryptosporidium Genotype in Pigs

    PubMed Central

    Ryan, U. M.; Samarasinghe, B.; Read, C.; Buddle, J. R.; Robertson, I. D.; Thompson, R. C. A.

    2003-01-01

    Over a 3-year period, a total of 646 fecal samples from pigs in 22 indoor and outdoor herds from Western Australia were screened for Cryptosporidium spp. by microscopy. Results revealed that 39 of 646 samples (6.03%) were positive for Cryptosporidium. Cryptosporidium was much more common in outdoor herds (17.2%) than in indoor herds (0.5%) and was more common in animals between the ages of 5 and 8 weeks (69.2%) than in younger animals (P < 0.0001). Molecular characterization of the positive samples at the 18S ribosomal DNA locus identified two distinct genotypes of Cryptosporidium: the previously identified pig genotype I and a novel pig genotype (pig genotype II), both of which warrant species status. PMID:12839769

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  1. Molecular-Based Detection Systems for Cryptosporidium Oocysts

    EPA Science Inventory

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

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

    EPA Science Inventory

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

  3. 40 CFR 141.718 - Treatment performance toolbox components.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for... performance. The State may approve Cryptosporidium treatment credit for drinking water treatment processes... 40 Protection of Environment 22 2010-07-01 2010-07-01 false Treatment performance...

  4. 40 CFR 141.718 - Treatment performance toolbox components.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ...) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for... performance. The State may approve Cryptosporidium treatment credit for drinking water treatment processes... 40 Protection of Environment 24 2013-07-01 2013-07-01 false Treatment performance...

  5. 40 CFR 141.718 - Treatment performance toolbox components.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ...) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for... performance. The State may approve Cryptosporidium treatment credit for drinking water treatment processes... 40 Protection of Environment 23 2014-07-01 2014-07-01 false Treatment performance...

  6. 40 CFR 141.718 - Treatment performance toolbox components.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for... performance. The State may approve Cryptosporidium treatment credit for drinking water treatment processes... 40 Protection of Environment 24 2012-07-01 2012-07-01 false Treatment performance...

  7. 40 CFR 141.718 - Treatment performance toolbox components.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...) WATER PROGRAMS (CONTINUED) NATIONAL PRIMARY DRINKING WATER REGULATIONS Enhanced Treatment for... performance. The State may approve Cryptosporidium treatment credit for drinking water treatment processes... 40 Protection of Environment 23 2011-07-01 2011-07-01 false Treatment performance...

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

    PubMed

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

    2015-01-01

    The objectives of this cross-sectional study were to detect the presence of Cryptosporidium spp. and Giardia duodenalis in drinking water treatments plants (DWTPs) in Galicia (NW Spain) and to identify which species and genotype of these pathogenic protozoans are present in the water. Samples of untreated water (surface or ground water sources) and of treated drinking water (in total, 254 samples) were collected from 127 DWTPs and analysed by an immunofluorescence antibody test (IFAT) and by PCR. Considering the untreated water samples, Cryptosporidium spp. were detected in 69 samples (54.3%) by IFAT, and DNA of this parasite was detected in 57 samples (44.8%) by PCR, whereas G. duodenalis was detected in 76 samples (59.8%) by IFAT and in 56 samples (44.0%) by PCR. Considering the treated drinking water samples, Cryptosporidium spp. was detected in 52 samples (40.9%) by IFAT, and the parasite DNA was detected in 51 samples (40.1%) by PCR, whereas G. duodenalis was detected in 58 samples (45.6%) by IFAT and in 43 samples (33.8%) by PCR. The percentage viability of the (oo)cysts ranged between 90.0% and 95.0% in all samples analysed. Cryptosporidium andersoni, C. hominis, C. parvum and assemblages A-I, A-II, E of G. duodenalis were identified. The results indicate that Cryptosporidium spp. and G. duodenalis are widespread in the environment and that DWTPs are largely ineffective in reducing/inactivating these pathogens in drinking water destined for human and animal consumption in Galicia. In conclusion, the findings suggest the need for better monitoring of water quality and identification of sources of contamination. PMID:25270421

  9. Quantitative analysis of Cryptosporidium growth in in vitro culture--the impact of parasite density on the success of infection.

    PubMed

    Paziewska-Harris, Anna; Singer, Martin; Schoone, Gerard; Schallig, Henk

    2016-01-01

    Cryptosporidium is an important waterborne pathogen for which no treatment or vaccination is available. This study set out to quantify DNA replication of Cryptosporidium parvum in vitro. Cryptosporidium DNA could be detected at up to 60 % of input level in both host-cell-free and host cell containing cultures 6 days after infection with living sporozoites, but was lost within 2 days in cultures inoculated with UV-inactivated sporozoites. Total DNA increased between days 2 and 6, evidence of successful DNA replication in both cell-free and host-cell-containing cultures. Overall however, only a small fraction (up to 5 %) of parasite DNA could be found associated with host cells or bound to plastic of the cell-free cultures, and the majority of parasite DNA was present in the cell culture medium, separable by simple decantation. After 2 days, in host-cell-containing cultures, the parasite DNA could be concentrated by slow centrifugation, suggesting that it was associated with intact parasite cells, but at 6 days, the majority could not be centrifuged and is therefore thought to have represented copies associated with dead and degraded parasites. In cell-free cultures and in larger plates, the majority of DNA was in this form. Performance of the parasite was best in small culture plates, and least in the largest plate sizes. We interpret these results as suggesting that Cryptosporidium sporozoites first bind to the host cell monolayer or to the plasticware, but then by 2 days, there has been a substantial release of parasites back into the medium. Host-cell-free cultures also supported modest replication and may have represented DNA synthesis in cells beginning merogony. The role of the host cells is unclear, as so much of the parasite DNA is released into the medium. Host cells may provide a feeder role, conditioning the medium for Cryptosporidium development. PMID:26435485

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

    PubMed Central

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

    2011-01-01

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

  11. A longitudinal study on the occurrence of Cryptosporidium and Giardia in dogs during their first year of life

    PubMed Central

    Hamnes, Inger S; Gjerde, Bjørn K; Robertson, Lucy J

    2007-01-01

    Background The primary aim of this study was to obtain more knowledge about the occurrence of Cryptosporidium and Giardia in young dogs in Norway. The occurrence of these parasites was investigated in a longitudinal study by repeated faecal sampling of dogs between 1 and 12 months of age (litter samples and individual samples). The dogs were privately owned and from four large breeds. Individual faecal samples were collected from 290 dogs from 57 litters when the dogs were approximately 3, 4, 6, and 12 months old. In addition, pooled samples were collected from 43 of the litters, and from 42 of the mother bitches, when the puppies were approximately 1 and/or 2 months old. Methods The samples were purified by sucrose gradient flotation concentration and examined by immunofluorescent staining. Results 128 (44.1%) of the young dogs had one or more Cryptosporidium positive samples, whilst 60 (20.7%) dogs had one or more Giardia positive samples. The prevalence of the parasites varied with age. For Cryptosporidium, the individual prevalence was between 5.1% and 22.5%, with the highest level in dogs < 6 months old, and declining with age. For Giardia, the individual prevalence was between 6.0% and 11.4%, with the highest level in dogs > 6 months old, but the differences between age groups were not statistically significant. Significant differences in prevalences were found in relation to geographic location of the dogs. Both parasites occurred at low prevalences in Northern Norway. Conclusion Both Cryptosporidium and Giardia are common in Norwegian dogs, with Cryptosporidium more prevalent than Giardia. Prevalences of the parasites were found to be influenced by age, geographical location, and infection status before weaning. PMID:17848186

  12. Treatment-resistant depression in adolescents: is the addition of cognitive behavioral therapy of benefit?

    PubMed Central

    Hetrick, Sarah E; Cox, Georgina R; Merry, Sally N

    2011-01-01

    Background Many young people with major depression fail first-line treatments. Treatment-resistant depression has various definitions in the literature but typically assumes nonresponse to medication. In young people, cognitive behavioral therapy (CBT) is the recommended first-line intervention, thus the definition of treatment resistance should be expanded. Therefore, our aim was to synthesize the existing evidence of any interventions for treatment-resistant depression, broadly defined, in children and adolescents and to investigate the effectiveness of CBT in this context. Methods We used Cochrane Collaboration methodology, with electronic searches of Medline, PsycINFO, Embase, and the Cochrane Depression Anxiety and Neurosis Group trials registers. Only randomized controlled trials were included, and were assessed for risk of bias. Meta- analysis was undertaken where possible and appropriate. Results Of 953 articles retrieved, four trials were eligible for inclusion. For one study, only the trial registration document was available, because the study was never completed. All other studies were well conducted with a low risk of bias, although one study had a high dropout rate. Two studies assessed the effect of adding CBT to medication. While an assertive trial of antidepressants does appear to lead to benefit, when compared with placebo, there was no significant advantage, in either study, or in a meta-analysis of data from these trials, that clearly demonstrated an additional benefit of CBT. The third trial showed little advantage of a tricyclic antidepressant over placebo in the context of an inpatient admission. Conclusion Few randomized controlled trials have investigated interventions for treatment-resistant depression in young people, and results from these show modest benefit from antidepressants with no additional benefit over medication from CBT. Overall, there is a lack of evidence about effective interventions to treat young people who have failed to

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

    PubMed Central

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

    2015-01-01

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

  14. Cryptosporidium ryanae n. sp. (Apicomplexa: Cryptosporidiidae) in cattle (Bos taurus).

    PubMed

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

    2008-10-01

    A new species, Cryptosporidium ryanae, is described from cattle. Oocysts of C. ryanae, previously identified as the Cryptosporidium deer-like genotype and recorded as such in GenBank (AY587166, EU203216, DQ182597, AY741309, and DQ871345), are similar to those of Cryptosporidium parvum and Cryptosporidium bovis but smaller. This genotype has been reported to be prevalent in cattle worldwide. Oocysts obtained from a calf for the present study are the smallest Cryptosporidium oocysts reported in mammals, measuring 2.94-4.41micromx2.94-3.68microm (mean=3.16micromx3.73microm) with a length/width shape index of 1.18 (n=40). The pre-patent period for two Cryptosporidium-naïve calves fed C. ryanae oocysts was 11 days and the patent period was 15-17 days. Oocysts were not infectious for BALB/c mice or lambs. Fragments of the SSU-rDNA, HSP-70, and actin genes amplified by PCR were purified and PCR products were sequenced. Multi-locus analysis of the three unlinked loci demonstrated the new species to be distinct from all other species and also demonstrated a lack of recombination, providing further evidence of species status. Based on morphological, molecular and biological data, this geographically widespread parasite found only in Bos taurus calves is recognized as a new species and is named C. ryanae. PMID:18583057

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

    PubMed

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

    2014-03-17

    The morphological, biological, and molecular characteristics of Cryptosporidium hedgehog genotype are described, and the species name Cryptosporidium erinacei n. sp. is proposed to reflect its specificity for hedgehogs under natural and experimental conditions. Oocysts of C. erinacei are morphologically indistinguishable from Cryptosporidium parvum, measuring 4.5-5.8 μm (mean=4.9 μm) × 4.0-4.8 μm (mean=4.4 μm) with a length to width ratio of 1.13 (1.02-1.35) (n=100). Oocysts of C. erinacei obtained from a naturally infected European hedgehog (Erinaceus europaeus) were infectious for naïve 8-week-old four-toed hedgehogs (Atelerix albiventris); the prepatent period was 4-5 days post infection (DPI) and the patent period was longer than 20 days. C. erinacei was not infectious for 8-week-old SCID and BALB/c mice (Mus musculus), Mongolian gerbils (Meriones unguiculatus), or golden hamsters (Mesocricetus auratus). Phylogenetic analyses based on small subunit rRNA, 60 kDa glycoprotein, actin, Cryptosporidium oocyst wall protein, thrombospondin-related adhesive protein of Cryptosporidium-1, and heat shock protein 70 gene sequences revealed that C. erinacei is genetically distinct from previously described Cryptosporidium species. PMID:24529828

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

    PubMed

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

    2013-06-01

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

  17. Polycyclic aromatic hydrocarbon removal from petroleum sludge cake using thermal treatment with additives.

    PubMed

    Pakpahan, Edward Nixon; Isa, Mohamed Hasnain; Kutty, Shamsul Rahman Mohamed; Chantara, Somporn; Wiriya, Wan

    2013-01-01

    Petroleum sludge is a hazardous waste that contains various organic compounds including polycyclic aromatic hydrocarbons (PAHs) which have carcinogenic-mutagenic and toxic characteristics. This study focuses on the thermal treatment (indirect heating) of petroleum sludge cake for PAH degradation at 250, 450, and 650 degrees C using Ca(OH)2 + NaHCO3 as an additive. The treatment was conducted in a rotary drum electric heater. All experiments were carried out in triplicate. Concentrations of the 16 priority PAHs in gas (absorbed on Amberlite XAD-4 adsorbent), particulate (on quartz filter) and residue phases were determined using gas chromatography-mass spectrometry (GC-MS). The samples were extracted with acetonitrile by ultra-sonication prior to GC-MS analysis. The use of additive was beneficial and a temperature of 450 degrees C was suitable for PAH degradation. Low levels of PAH emissions, particularly carcinogenic PAH and toxic equivalent concentration (sigma TEC), were observed in gas, particulate and residue phases after treatment. PMID:23530354

  18. Investigating Attachment Behaviors of Cryptosporidium Parvum Oocysts Using Collision Efficiency in Laboratory Column Experiments

    NASA Astrophysics Data System (ADS)

    Park, Y.; Hou, L.; Atwill, R.; Packman, A. I.; Harter, T.

    2009-12-01

    Cryptosporidium is one of the most common enteric parasites of humans and domestic animals, and a number of outbreaks of Cryprosporidiosis, a diarrheal disease caused by Cryptosporidium have been reported worldwide. Natural porous media has been demonstrated to be an effective filter for removing Cryptosporidium parvum from contaminated water and the amount of Cryptosporidium filtered is known to be highly dependent on physical and chemical conditions of the porous media and the water. Cryptosporidium deposition in saturated porous media involves two main steps: approach and attachment. In contrast to the approach mechanisms, attachment processes have not been systematically described to predict a priori because theories that represent attachment behavior (colloid stability) such as DLVO are insufficient to explain experimental data. For this reason, attachment efficiency is calculated based on empirical data, typically experimental breakthrough curves in laboratory columns or field experiments. In this study, collision (attachment) efficiencies (α) of C. parvum oocyst were calculated to test the effect of chemical property changes on the association of oocysts with sand grains. The breakthrough curve data obtained from twelve column experiments and three models were employed to calculate single collector efficiency (η) and α. The first ten experiments were conducted by changing ionic strength and pH, and mixing with natural sediments under the same physical properties (same η). Our experiment results show that iron coating or clay/suspended solids mixture drastically enhanced oocyst deposition. The experiments also showed that increase in ionic strength and decrease in pH enhanced the attachment efficiency. However, the experiment with 100mM NaCl resulted in low attachment efficiency and the experiment with pH 8.5 showed similar attachment efficiency to the one at pH 7. Based on the results from two additional experiments with different flow velocities, it

  19. Effect of high hydrostatic pressure on Cryptosporidium parvum infectivity.

    PubMed

    Slifko, T R; Raghubeer, E; Rose, J B

    2000-09-01

    The incidence of foodborne disease outbreaks caused by contaminated low-pH fruit juices is increasing. With recent mandatory pasteurization of apple juice and the industry's concerns of food safety, fruit juice processors are showing more interest in alternative nonthermal technologies that can kill >99.99% of microbial pathogens present in foods. The association of the coccidian protozoan, Cryptosporidium, with diarrheal disease outbreaks from contaminated tap water and fruit juice raises a safety concern in the food and beverage industries. The objective of this study was to evaluate the effects of high hydrostatic pressure (HHP) on C. parvum oocysts. Oocysts were suspended in apple and orange juice and HHP treated at 5.5 x 10(8) Pa (80,000 psi) for 0, 30, 45, 60, 90, and 120 s. Oocyst viability was assessed by excystation using bile salts and trypsin while the cell culture foci detection method was used to assess infectivity. Results indicated that HHP inactivated C. parvum oocysts by at least 3.4 log10 after 30 s of treatment. No infectivity was detected in samples exposed to > or =60 s of HHP and >99.995% inactivation was observed. This study demonstrated that HHP efficiently rendered the oocysts nonviable and noninfectious after treatment at 5.5 x 10(8) Pa. PMID:10983803

  20. The Facial Aesthetic index: An additional tool for assessing treatment need

    PubMed Central

    Sundareswaran, Shobha; Ramakrishnan, Ranjith

    2016-01-01

    Objectives: Facial Aesthetics, a major consideration in orthodontic diagnosis and treatment planning, may not be judged correctly and completely by simply analyzing dental occlusion or osseous structures. Despite this importance, there is no index to guarantee availability of treatment or prioritize patients based on their soft tissue treatment needs. Individuals having well-aligned teeth but unaesthetic convex profiles do not get included for treatment as per current malocclusion indices. The aim of this investigation is to develop an aesthetic index based on facial profiles which could be used as an additional tool with malocclusion indices. Materials and Methods: A chart showing typical facial profile changes due to underlying malocclusions was generated by soft tissue manipulations of standardized profile photographs of a well-balanced male and female face. A panel of 62 orthodontists judged the profile photographs of 100 patients with different soft tissue patterns for assessing profile variations and treatment need. The index was later tested in a cross-section of school population. Statistical analysis was done using “irr” package of R environment version 2.15.1. Results: The index exhibited very good reliability in determining profile variations (Fleiss kappa 0.866, P < 0.001), excellent reproducibility (kappa 0.9078), high sensitivity, and specificity (95.7%). Testing in population yielded excellent agreement among orthodontists (kappa 0.9286). Conclusions: A new Facial Aesthetic index, based on patient's soft tissue profile requirements is proposed, which can complement existing indices to ensure treatment to those in need. PMID:27127752

  1. Effect of organic acids and hydrogen peroxide on Cryptosporidium parvum viability in fruit juices.

    PubMed

    Kniel, Kalmia E; Sumner, Susan S; Lindsay, David S; Hackney, Cameron R; Pierson, Merle D; Zajac, Anne M; Golden, David A; Fayer, Ronald

    2003-09-01

    Cryptosporidium parvum has historically been associated with waterborne outbreaks of diarrheal illness. Foodborne cryptosporidiosis has been associated with unpasteurized apple cider. Infectious oocysts are shed in the feces of common ruminants like cattle and deer in and near orchards. In this study, the ability of organic acids and hydrogen peroxide (H2O2) added to fruit juice to inhibit the survival of C. parvum was analyzed. Oocyst viability was analyzed by a cell culture infectivity assay with the use of a human ileocecal cell line (HCT-8) whose infectivity pattern is similar to that for human oral infectivity. Cell monolayers were infected with 10(6) treated oocysts or a series of 10-fold dilutions. Parasitic life stages were visualized through immunohistochemistry with 100 microscope fields per monolayer being counted. In vitro excystation assays were also used to evaluate these treatments. Organic acids and H2O2 were added to apple cider, orange juice, and grape juices on a weight/volume basis. Malic, citric, and tartaric acids at concentrations of 1 to 5% inhibited C. parvum's infectivity of HCT-8 cells by up to 88%. Concentrations ranging from 0.025 to 3% H2O2 were evaluated. The addition of 0.025% H2O2 to each juice resulted in a >5-log reduction of C. parvum infectivity as determined with a most-probable-number-based cell culture infectivity assay. As observed with differential interference contrast and scanning electron microscopy, reduced infectivity may be mediated through effects on the oocyst wall that are caused by the action of H2O2 or related oxygen radicals. The addition of low concentrations of H2O2 can represent a valuable alternative to pasteurization. PMID:14503720

  2. Additive protection by LDR and FGF21 treatment against diabetic nephropathy in type 2 diabetes model

    PubMed Central

    Shao, Minglong; Yu, Lechu; Zhang, Fangfang; Lu, Xuemian; Li, Xiaokun; Cheng, Peng; Lin, Xiufei; He, Luqing; Jin, Shunzi; Tan, Yi; Yang, Hong; Cai, Lu

    2015-01-01

    The onset of diabetic nephropathy (DN) is associated with both systemic and renal changes. Fibroblast growth factor (FGF)-21 prevents diabetic complications mainly by improving systemic metabolism. In addition, low-dose radiation (LDR) protects mice from DN directly by preventing renal oxidative stress and inflammation. In the present study, we tried to define whether the combination of FGF21 and LDR could further prevent DN by blocking its systemic and renal pathogeneses. To this end, type 2 diabetes was induced by feeding a high-fat diet for 12 wk followed by a single dose injection of streptozotocin. Diabetic mice were exposed to 50 mGy LDR every other day for 4 wk with and without 1.5 mg/kg FGF21 daily for 8 wk. The changes in systemic parameters, including blood glucose levels, lipid profiles, and insulin resistance, as well as renal pathology, were examined. Diabetic mice exhibited renal dysfunction and pathological abnormalities, all of which were prevented significantly by LDR and/or FGF21; the best effects were observed in the group that received the combination treatment. Our studies revealed that the additive renal protection conferred by the combined treatment against diabetes-induced renal fibrosis, inflammation, and oxidative damage was associated with the systemic improvement of hyperglycemia, hyperlipidemia, and insulin resistance. These results suggest that the combination treatment with LDR and FGF21 prevented DN more efficiently than did either treatment alone. The mechanism behind these protective effects could be attributed to the suppression of both systemic and renal pathways. PMID:25968574

  3. Identification of algae which interfere with the detection of Giardia cysts and Cryptosporidium oocysts and a method for alleviating this interference.

    PubMed Central

    Rodgers, M R; Flanigan, D J; Jakubowski, W

    1995-01-01

    Fifty-four algal species were tested for cross-reaction in the American Society for Testing and Materials Giardia/Cryptosporidium indirect immunofluorescence assay, and 24 showed some degree of fluorescence. Two species, Navicula minima and Synechococcus elongatus, exhibited a bright apple green fluorescence. The addition of goat serum to the assay mixture blocked the fluorescence of most nontarget organisms tested and also decreased the background fluorescence. Goat serum did not interfere with the fluorescence of Giardia cysts or Cryptosporidium oocysts or the identification of cyst and oocyst internal structures. PMID:7487013

  4. A novel multiplex PCR coupled with Luminex assay for the simultaneous detection of Cryptosporidium spp., Cryptosporidium parvum and Giardia duodenalis.

    PubMed

    Li, Wei; Zhang, Nan; Gong, Pengtao; Cao, Lili; Li, Jianhua; Su, Libo; Li, Shuhong; Diao, Yumei; Wu, Kang; Li, He; Zhang, Xichen

    2010-10-11

    Cryptosporidium parvum and Giardia duodenalis are the most frequently identified enteric parasites associated with diarrhea-causing disease outbreaks, and many non-parvum species of Cryptosporidium also can replicate and cause illness in mammals including humans. In this study, we describe a novel multiplex PCR coupled with Luminex assay for the identification of Cryptosporidium spp., C. parvum and G. duodenalis in a rapid manner. The multiplex PCR for the simultaneous detection of Cryptosporidium and Giardia was developed using three pairs of biotinylated primers which amplify 424, 223 and 267 bp products from the U1 small nuclear ribonucleoprotein (U1 snr) gene, 18S rRNA gene of Cryptosporidium and the beta-giardin gene of Giardia, respectively. The genus and species-specific capture probes linked to carboxylated Luminex microspheres hybridized to the multiplex PCR amplicons to enhance sensitivity and specificity. The conditions of multiplex PCR and Luminex hybridization reaction were optimized to enable the minimum detection limits of 5x10(-6), 5x10(-6), and 5x10(-6) ng DNAs (corresponding approximately to 0.1 oocyst/cyst). The Luminex approach proved to be 100% specific and accurate by testing a total of 240 fecal samples compared with microscopic examination of fecal smears and further modified acid-fast staining or iodine-staining observation. The established assay offers the potential for rapid detection of Cryptosporidium spp., C. parvum and G. duodenalis in fecal and environmental samples. PMID:20594647

  5. In Vitro Cultivation of Cryptosporidium Species

    PubMed Central

    Arrowood, Michael J.

    2002-01-01

    The in vitro cultivation of protozoan parasites of the genus Cryptosporidium has advanced significantly in recent years. These obligate, intracellular parasites colonize the epithelium of the digestive and respiratory tracts, are often difficult to obtain in significant numbers, produce durable oocysts that defy conventional chemical disinfection methods, and are persistently infectious when stored at refrigerated temperatures (4 to 8°C). While continuous culture and efficient life cycle completion (oocyst production) have not yet been achieved in vitro, routine methods for parasite preparation and cell culture infection and assays for parasite life cycle development have been established. Parasite yields may be limited, but in vitro growth is sufficient to support a variety of research studies, including assessing potential drug therapies, evaluating oocyst disinfection methods, and characterizing life cycle stage development and differentiation. PMID:12097247

  6. Cryptosporidium: detection in water and food.

    PubMed

    Smith, Huw V; Nichols, Rosely A B

    2010-01-01

    Water and food are major environmental transmission routes for Cryptosporidium, but our ability to identify the spectrum of oocyst contributions in current performance-based methods is limited. Determining risks in water and foodstuffs, and the importance of zoonotic transmission, requires the use of molecular methods, which add value to performance-based morphologic methods. Multi-locus approaches increase the accuracy of identification, as many signatures detected in water originate from species/genotypes that are not infectious to humans. Method optimisation is necessary for detecting small numbers of oocysts in environmental samples consistently, and further work is required to (i) optimise IMS recovery efficiency, (ii) quality assure performance-based methods, (iii) maximise DNA extraction and purification, (iv) adopt standardised and validated loci and primers, (v) determine the species and subspecies range in samples containing mixtures, and standardising storage and transport matrices for validating genetic loci, primer sets and DNA sequences. PMID:19501088

  7. Infection status of pigs with Cryptosporidium parvum

    PubMed Central

    Yu, Jae-Ran

    2004-01-01

    To investigate the infection status of pigs with Cryptosporidium parvum, 589 fecal samples were collected from pigs raised at farm in Chungcheongbuk-do and Chungcheongnam-do. Of the 589 pig fecal samples, 62 (10.5%) were positive for C. parvum. The area showing the highest positive rate was Dangjin-gun, Chungcheongnam-do (14.0%), and the lowest (0%) Salmi-myon, Chungcheongbuk-do. The positive rate of C. parvum in Judok-eup increased from 12.7% in the winter to 22.1% in the summer. The results of this study suggest that the pigs may be a source of human C. parvum infection. PMID:15060340

  8. Improved Risk Analysis by Dual Direct Detection of Total and Infectious Cryptosporidium Oocysts on Cell Culture in Combination with Immunofluorescence Assay▿

    PubMed Central

    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

  9. ETV STATEMENT - PHYSICAL REMOVAL OF CRYPTOSPORIDIUM OOCYSTS AND GIARDIA CYSTS IN DRINKING WATER - ZENON ZEEWEED ZW500 ULTRAFILTRATION MEMBRANE SYSTEM AT PITTSBURGH, PA

    EPA Science Inventory

    Verification testing of the ZENON Environmental Inc. ZeeWeed ZW500 UF Drinking Water System was conducted from 2/6-3/7/99. The treatment system underwent Giardia and Cryptosporidium removal challenge testing on 3/2/99 and demonstrated a 5.3 log10 removal of Giardia cysts and a 6...

  10. Effects of thermal treatments on the characterisation and utilisation of red mud with sawdust additive.

    PubMed

    Liu, Yanju; Naidu, Ravi; Ming, Hui; Dharmarajan, Rajarathnam; Du, Jianhua

    2016-06-01

    Extremely large amounts of red mud (bauxite residue) are generated globally every year from alumina refining industries, which are being disposed of on engineered landfills. The objective of this study is to investigate the effects of thermal treatments on red mud for development of utilisation strategies. Thermal treatments of red mud samples and their characterisations were investigated under inert (N2) and oxidative (air) conditions with and without sawdust addition at 200-600°C. After calcination, the resulting samples were analysed using thermogravimetric-infrared spectroscopy (TG-IR) for functional group transformations, thermogravimetric analysis (TGA) for thermal loss profiles and X-ray diffraction (XRD) for mineral transformations. The characterisation results showed that in N2 environment, boehmite in red mud was transferred to transition alumina at around 400°C while losing water from structural components. The addition of sawdust for incubation and calcination of red mud in air increased the surface area, whereas that in nitrogen atmosphere lead to reduction of hematite to magnetite at around 500°C. The incorporated carbon materials played a major role in increasing the surface area especially for pore size less than 2.5 nm. This treated red mud with altered mineral composition and improved properties for binding contaminants can be used for environmental remediation and in the process of metal recovery such as iron. PMID:26951343

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

    PubMed

    Liu, Xuehan; He, Tingmei; Zhong, Zhijun; Zhang, Hemin; Wang, Rongjun; Dong, Haiju; Wang, Chengdong; Li, Desheng; Deng, Jiabo; Peng, Guangneng; Zhang, Longxian

    2013-10-01

    Fifty-seven fecal samples were collected from giant pandas (Ailuropoda melanoleuca) in the China Conservation and Research Centre for the Giant Panda (CCRCGP) in Sichuan and examined for Cryptosporidium oocysts by Sheather's sugar flotation technique. An 18-year-old male giant panda was Cryptosporidium positive, with oocysts of an average size of 4.60×3.99 μm (n=50). The isolate was genetically analyzed using the partial 18S rRNA, 70 kDa heat shock protein (HSP70), Cryptosporidium oocyst wall protein (COWP) and actin genes. Multi-locus genetic characterization indicated that the present isolate was different from known Cryptosporidium species and genotypes. The closest relative was the Cryptosporidium bear genotype, with 11, 10, and 6 nucleotide differences in the 18S rRNA, HSP70, and actin genes, respectively. Significant differences were also observed in the COWP gene compared to Cryptosporidium mongoose genotype. The homology to the bear genotype at the 18S rRNA locus was 98.6%, which is comparable to that between Cryptosporidium parvum and Cryptosporidium hominis (99.2%), or between Cryptosporidium muris and Cryptosporidium andersoni (99.4%). Therefore, the Cryptosporidium in giant pandas in this study is considered as a new genotype: the Cryptosporidium giant panda genotype. PMID:23810821

  12. USE OF LONG ACTING STEROID METHYLPREDNISOLONE ACETATE FOR THE PROLONGATION OF CRYPTOSPORIDIUM SP. IN MICE

    EPA Science Inventory

    Current protocols for Cryptosporidium sp. propagation in mice vary according to the strain and age of mouse as well as the species of Cryptosporidium. Cryptosporidium muris is a natural parasite of mice, but only neonatal mice are susceptible to C. parvum infection. Published C...

  13. A multiplex PCR to simultaneously distinguish Cryptosporidium species of veterinary and public health concern in cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Four species of Cryptosporidium are routinely found in cattle: Cryptosporidium parvum, C. bovis, C. ryanae, and C. andersoni. It is important to determine the species of Cryptosporidium in infected cattle because C. parvum is the only serious pathogen, for humans as well as cattle. Identification of...

  14. Nitrification in lake sediment with addition of drinking water treatment residuals.

    PubMed

    Wang, Changhui; Liu, Juanfeng; Wang, Zhixin; Pei, Yuansheng

    2014-06-01

    Drinking water treatment residuals (WTRs), non-hazardous by-products generated during potable water production, can effectively reduce the lake internal phosphorus (P) loading and improve water quality in lakes. It stands to reason that special attention regarding the beneficial reuse of WTRs should be given not only to the effectiveness of P pollution control, but also to the effects on the migration and transformation of other nutrients (e.g., nitrogen (N)). In this work, based on laboratory enrichment tests, the effects of WTRs addition on nitrification in lake sediment were investigated using batch tests, fluorescence in situ hybridization, quantitative polymerase chain reaction and phylogenetic analysis techniques. The results indicated that WTRs addition had minor effects on the morphologies of AOB and NOB; however, the addition slightly enhanced the sediment nitrification potential from 12.8 to 13.2 μg-N g(-1)-dry sample h(-1) and also increased the ammonia oxidation bacteria (AOB) and nitrite oxidizing bacteria (NOB) abundances, particularly the AOB abundances (P < 0.05), which increased from 1.11 × 10(8) to 1.31 × 10(8) copies g(-1)-dry sample. Moreover, WTRs addition was beneficial to the enrichment of Nitrosomonas and Nitrosospira multiformis and promoted the emergence of a new Nitrospira cluster, causing the increase in AOB and NOB diversities. Further analysis showed that the variations of nitrification in lake sediment after WTRs addition were primarily due to the decrease of bioavailable P, the introduction of new nitrifiers and the increase of favorable carriers for microorganism attachment in sediments. Overall, these results suggested that WTRs reuse for the control of lake internal P loading would also lead to conditions that are beneficial to nitrification. PMID:24681379

  15. Addition of meloxicam to the treatment of clinical mastitis improves subsequent reproductive performance.

    PubMed

    McDougall, S; Abbeloos, E; Piepers, S; Rao, A S; Astiz, S; van Werven, T; Statham, J; Pérez-Villalobos, N

    2016-03-01

    A blinded, negative controlled, randomized intervention study was undertaken to test the hypothesis that addition of meloxicam, a nonsteroidal anti-inflammatory drug, to antimicrobial treatment of mild to moderate clinical mastitis would improve fertility and reduce the risk of removal from the herd. Cows (n=509) from 61 herds in 8 regions (sites) in 6 European countries were enrolled. Following herd-owner diagnosis of mild to moderate clinical mastitis within the first 120 d of lactation in a single gland, the rectal temperature, milk appearance, and California Mastitis Test score were assessed. Cows were randomly assigned within each site to be treated either with meloxicam or a placebo (control). All cows were additionally treated with 1 to 4 intramammary infusions of cephalexin and kanamycin at 24-h intervals. Prior to treatment and at 14 and 21 d posttreatment, milk samples were collected for bacteriology and somatic cell count. Cows were bred by artificial insemination and pregnancy status was subsequently defined. General estimating equations were used to determine the effect of treatment (meloxicam versus control) on bacteriological cure, somatic cell count, the probability of being inseminated by 21 d after the voluntary waiting period, the probability of conception to first artificial insemination, the number of artificial insemination/conception, the probability of pregnancy by 120 or 200 d postcalving, and the risk of removal by 300 d after treatment. Cox's proportional hazards models were used to test the effect of treatment on the calving to first insemination and calving to conception intervals. Groups did not differ in terms of age, clot score, California Mastitis Test score, rectal temperature, number of antimicrobial treatments given or bacteria present at the time of enrollment, but cows treated with meloxicam had greater days in milk at enrollment. Cows treated with meloxicam had a higher bacteriological cure proportion than those treated with

  16. Effectiveness of Standard UV Depuration at Inactivating Cryptosporidium parvum Recovered from Spiked Pacific Oysters (Crassostrea gigas)▿

    PubMed Central

    Sunnotel, O.; Snelling, W. J.; McDonough, N.; Browne, L.; Moore, J. E.; Dooley, J. S. G.; Lowery, C. J.

    2007-01-01

    When filter-feeding shellfish are consumed raw, because of their ability to concentrate and store waterborne pathogens, they are being increasingly associated with human gastroenteritis and have become recognized as important pathogen vectors. In the shellfish industry, UV depuration procedures are mandatory to reduce pathogen levels prior to human consumption. However, these guidelines are based around more susceptible fecal coliforms and Salmonella spp. and do not consider Cryptosporidium spp., which have significant resistance to environmental stresses. Thus, there is an urgent need to evaluate the efficiency of standard UV depuration against the survival of Cryptosporidium recovered from shellfish. Our study found that in industrial-scale shellfish depuration treatment tanks, standard UV treatment resulted in a 13-fold inactivation of recovered, viable C. parvum oocysts from spiked (1 × 106 oocysts liter −1) Pacific oysters. Depuration at half power also significantly reduced (P < 0.05; ninefold) the number of viable oocysts recovered from oysters. While UV treatment resulted in significant reductions of recovered viable oocysts, low numbers of viable oocysts were still recovered from oysters after depuration, making their consumption when raw a public health risk. Our study highlights the need for increased periodic monitoring programs for shellfish harvesting sites, improved depuration procedures, and revised microbial quality control parameters, including Cryptosporidium assessment, to minimize the risk of cryptosporidiosis. PMID:17574996

  17. Inactivation of Bacillus subtilis spores using various combinations of ultraviolet treatment with addition of hydrogen peroxide.

    PubMed

    Zhang, Yiqing; Zhou, Lingling; Zhang, Yongji; Tan, Chaoqun

    2014-01-01

    This study aims at comparing the inactivation of Bacillus subtilis spores by various combinations of UV treatment and hydrogen peroxide (H2O2) addition. The combinations included sequential (UV-H2O2, H2O2-UV) and simultaneous (UV/H2O2) processes. Results showed that B. subtilis spores achieved a certain inactivation effect through UV treatment. However, hardly any inactivation effect by H2O2 alone was observed. H2O2 had a significant synergetic effect when combined with UV treatment, while high irradiance and H2O2 concentration both favored the reaction. When treated with 0.60 mm H2O2 and 113.0 μW/cm(2) UV irradiance for 6 min, the simultaneous UV/H2O2 treatment showed significantly improved disinfection effect (4.13 log) compared to that of UV-H2O2 (3.03 log) and H2O2-UV (2.88 log). The relationship between the inactivation effect and the exposure time followed a typical pseudo-first-order kinetics model. The pseudo-first-order rate constants were 0.478, 0.447 and 0.634 min(-1), for the UV-H2O2, H2O2-UV and UV/H2O2 processes, respectively, further confirming the optimal disinfection effect of the UV/H2O2 process. The disinfection could be ascribed to the OH radicals, as verified by the level of para-chlorobenzoic acid (pCBA). PMID:24447294

  18. Survival of Cryptosporidium parvum oocysts in the presence of hydrated lime.

    PubMed

    Zintl, A; Keogh, B; Ezzaty-Mirhashemi, M; De Waal, T; Scholz, D; Mulcahy, G

    2010-03-01

    To investigate the effects of hydrated lime on the survival of Cryptosporidium oocysts, the percentage viability of oocysts was assessed using fluorescent in situ hybridisation. In the absence of lime and with lime at a concentration of 1 per cent, there was a gradual decline in oocyst viability during the 10-day trial. Although the addition of 5 or 10 per cent lime caused the total number of oocysts to decrease, there appeared to be an increase in the proportion of potentially viable oocysts. PMID:20208077

  19. Drug Development Against the Major Diarrhea-Causing Parasites of the Small Intestine, Cryptosporidium and Giardia.

    PubMed

    Miyamoto, Yukiko; Eckmann, Lars

    2015-01-01

    Diarrheal diseases are among the leading causes of morbidity and mortality in the world, particularly among young children. A limited number of infectious agents account for most of these illnesses, raising the hope that advances in the treatment and prevention of these infections can have global health impact. The two most important parasitic causes of diarrheal disease are Cryptosporidium and Giardia. Both parasites infect predominantly the small intestine and colonize the lumen and epithelial surface, but do not invade deeper mucosal layers. This review discusses the therapeutic challenges, current treatment options, and drug development efforts against cryptosporidiosis and giardiasis. The goals of drug development against Cryptosporidium and Giardia are different. For Cryptosporidium, only one moderately effective drug (nitazoxanide) is available, so novel classes of more effective drugs are a high priority. Furthermore, new genetic technology to identify potential drug targets and better assays for functional evaluation of these targets throughout the parasite life cycle are needed for advancing anticryptosporidial drug design. By comparison, for Giardia, several classes of drugs with good efficacy exist, but dosing regimens are suboptimal and emerging resistance begins to threaten clinical utility. Consequently, improvements in potency and dosing, and the ability to overcome existing and prevent new forms of drug resistance are priorities in antigiardial drug development. Current work on new drugs against both infections has revealed promising strategies and new drug leads. However, the primary challenge for further drug development is the underlying economics, as both parasitic infections are considered Neglected Diseases with low funding priority and limited commercial interest. If a new urgency in medical progress against these infections can be raised at national funding agencies or philanthropic organizations, meaningful and timely progress is possible

  20. Drug Development Against the Major Diarrhea-Causing Parasites of the Small Intestine, Cryptosporidium and Giardia

    PubMed Central

    Miyamoto, Yukiko; Eckmann, Lars

    2015-01-01

    Diarrheal diseases are among the leading causes of morbidity and mortality in the world, particularly among young children. A limited number of infectious agents account for most of these illnesses, raising the hope that advances in the treatment and prevention of these infections can have global health impact. The two most important parasitic causes of diarrheal disease are Cryptosporidium and Giardia. Both parasites infect predominantly the small intestine and colonize the lumen and epithelial surface, but do not invade deeper mucosal layers. This review discusses the therapeutic challenges, current treatment options, and drug development efforts against cryptosporidiosis and giardiasis. The goals of drug development against Cryptosporidium and Giardia are different. For Cryptosporidium, only one moderately effective drug (nitazoxanide) is available, so novel classes of more effective drugs are a high priority. Furthermore, new genetic technology to identify potential drug targets and better assays for functional evaluation of these targets throughout the parasite life cycle are needed for advancing anticryptosporidial drug design. By comparison, for Giardia, several classes of drugs with good efficacy exist, but dosing regimens are suboptimal and emerging resistance begins to threaten clinical utility. Consequently, improvements in potency and dosing, and the ability to overcome existing and prevent new forms of drug resistance are priorities in antigiardial drug development. Current work on new drugs against both infections has revealed promising strategies and new drug leads. However, the primary challenge for further drug development is the underlying economics, as both parasitic infections are considered Neglected Diseases with low funding priority and limited commercial interest. If a new urgency in medical progress against these infections can be raised at national funding agencies or philanthropic organizations, meaningful and timely progress is possible

  1. Hot-Pack and 1-MHz Ultrasound Treatments Have an Additive Effect on Muscle Temperature Increase

    PubMed Central

    Draper, David O.; Harris, Shane T.; Schulthies, Shane; Durrant, Earlene; Knight, Kenneth L.; Ricard, Mark

    1998-01-01

    Objective: Therapeutic ultrasound is an effective deep heating modality commonly applied alone or after cooling or heating of the treatment area. The purpose of this study was to examine the tissue temperature rise in the human triceps surae muscle group after ultrasound with prior heating via a silicate gel hot pack. Design and Setting: This study was designed as a 2 × 2 × 3 factorial with repeated measures on two factors (depth and time). Independent variables were temperature of pack (hot and room temperature), depth of measurement (1 cm and 3 cm), and time (beginning, after pack application, and after ultrasound). The dependent variable was tissue temperature. Subjects were assigned to one of two treatment groups: ultrasound preceded by a 15-minute hot pack treatment or ultrasound preceded by a 15-minute application with a silicate gel pack at room temperature. Measurements were taken while subjects were treated in a university training room. Subjects: Twenty-one uninjured male and female college student volunteers were randomly assigned to one of the two pack groups. Measurements: The hot packs were stored in 75°C water. A 1-MHz ultrasound treatment was administered for 10 minutes at an intensity of 1.5 W/cm². Tissue temperature was measured every 30 seconds using 23-gauge hypodermic microprobes interfaced with a telethermometer and inserted 1 and 3 cm below the surface of anesthetized triceps surae muscle. Results: At both tissue depths, there was a 0.8°C greater increase in tissue temperature with hot packs and ultrasound. At 1 cm, ultrasound increased temperature 3.5°C after a 0.5°C rise during the room temperature-pack application, but only 0.6°C after a 3.8°C increase during hot-pack application. At 3 cm, ultrasound increased temperature 3.85°C following a slight (-0.26°C) decrease during the room temperature-pack application and 3.68°C after a 0.74°C increase during hot-pack application. Conclusions: Vigorous increases in deep muscle

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

    PubMed Central

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

    2014-01-01

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

  3. Transport of Cryptosporidium parvum Oocysts in a Silicon Micromodel

    SciTech Connect

    Liu, Yuanyuan; Zhang, Changyong; Hilpert, Markus; Kuhlenschmidt, Mark S.; Kuhlenschmidt, Theresa B.; Nguyen, Thanh H.

    2012-02-01

    Effective removal of Cryptosporidium parvum oocysts by granular filtration requires the knowledge of oocyst transport and deposition mechanisms, which can be obtained based on real time microscopic observation of oocyst transport in porous media. Attachment of oocysts to silica surface in a radial stagnation point flow (RSPF) cell and in a micromodel, which has 2-dimensional (2-D) microscopic pore structures consisting of an array of cylindrical collectors, was studied and compared. Real time transport of oocysts in the micromodel was recorded to determine the attached oocyst distributions in transversal and longitudinal directions. In the micromodel, oocysts attached to the forward portion of clean collectors, where the flow velocity was lowest. After initial attachment, oocysts attached onto already attached oocysts. As a result, the collectors ripened and the region available for flow was reduced. Results of attachment and detachment experiments suggest that surface charge heterogeneity allowed for oocyst attachment. In addition to experiments, Lattice-Boltzmann simulations helped understanding the slightly non-uniform flow field and explained differences in the removal efficiency in the transversal direction. However, the hydrodynamic modeling could not explain differences in attachment in the longitudinal direction.

  4. Treatment of enterococcus faecalis bacteria by a helium atmospheric cold plasma brush with oxygen addition

    SciTech Connect

    Chen Wei; Huang Jun; Wang Xingquan; Lv Guohua; Zhang Guoping; Du Ning; Liu Xiaodi; Guo Lihong; Yang Size

    2012-07-01

    An atmospheric cold plasma brush suitable for large area and low-temperature plasma-based sterilization is designed. Results demonstrate that the He/O{sub 2} plasma more effectively kills Enterococcus faecalis than the pure He plasma. In addition, the sterilization efficiency values of the He/O{sub 2} plasma depend on the oxygen fraction in Helium gas. The atmospheric cold plasma brush using a proper ratio of He/O{sub 2} (2.5%) reaches the optimum sterilization efficiency. After plasma treatment, the cell structure and morphology changes can be observed by the scanning electron microscopy. Optical emission measurements indicate that reactive species such as O and OH play a significant role in the sterilization process.

  5. Treatment of enterococcus faecalis bacteria by a helium atmospheric cold plasma brush with oxygen addition

    NASA Astrophysics Data System (ADS)

    Chen, Wei; Huang, Jun; Du, Ning; Liu, Xiao-Di; Wang, Xing-Quan; Lv, Guo-Hua; Zhang, Guo-Ping; Guo, Li-Hong; Yang, Si-Ze

    2012-07-01

    An atmospheric cold plasma brush suitable for large area and low-temperature plasma-based sterilization is designed. Results demonstrate that the He/O2 plasma more effectively kills Enterococcus faecalis than the pure He plasma. In addition, the sterilization efficiency values of the He/O2 plasma depend on the oxygen fraction in Helium gas. The atmospheric cold plasma brush using a proper ratio of He/O2 (2.5%) reaches the optimum sterilization efficiency. After plasma treatment, the cell structure and morphology changes can be observed by the scanning electron microscopy. Optical emission measurements indicate that reactive species such as O and OH play a significant role in the sterilization process.

  6. Exhaust after-treatment system with in-cylinder addition of unburnt hydrocarbons

    DOEpatents

    Coleman, Gerald N.; Kesse, Mary L.

    2007-10-30

    Certain exhaust after-treatment devices, at least periodically, require the addition of unburnt hydrocarbons in order to create reductant-rich exhaust conditions. The present disclosure adds unburnt hydrocarbons to exhaust from at least one combustion chamber by positioning, at least partially within a combustion chamber, a mixed-mode fuel injector operable to inject fuel into the combustion chamber in a first spray pattern with a small average angle relative to a centerline of the combustion chamber and a second spray pattern with a large average angle relative to the centerline of the combustion chamber. An amount of fuel is injected in the first spray pattern into a non-combustible environment within the at least one combustion chamber during at least one of an expansion stroke and exhaust stroke. The exhaust with the unburnt amount of fuel is moved into an exhaust passage via an exhaust valve.

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

    PubMed

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

    2015-02-01

    The occurrence of Giardia and Cryptosporidium was investigated in cetacean specimens stranded on the northwestern coast of Spain (European Atlantic coast) by analysis of 65 samples of large intestine from eight species. The parasites were identified by direct immunofluorescence antibody test (IFAT) and by PCR amplification of the β-giardin gene, the ITS1-5.8S-ITS2 region and the SSU-rDNA gene of Giardia and the SSU-rDNA gene of Cryptosporidium. Giardia and Cryptosporidium were detected in 7 (10.8 %) and 9 samples (13.8 %), respectively. In two samples, co-infection with both parasites was observed. Giardia duodenalis assemblages A, C, D and F, and Cryptosporidium parvum were identified. This is the first report of G. duodenalis in Balaenoptera acutorostrata, Kogia breviceps and Stenella coeruleoalba and also the first report of Cryptosporidium sp. in B. acutorostrata and of C. parvum in S. coeruleoalba and Tursiops truncatus. These results extend the known host range of these waterborne enteroparasites. PMID:25418072

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

    PubMed

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

    2014-10-15

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

  9. Infection and diarrhea caused by Cryptosporidium sp. among Guatemalan infants.

    PubMed Central

    Cruz, J R; Cano, F; Càceres, P; Chew, F; Pareja, G

    1988-01-01

    During July 1985 to June 1986, fecal excretion of Cryptosporidium oocysts was determined prospectively in a cohort of 130 infants, aged 0 to 11 months, living in a marginal urban area of Guatemala City, Guatemala. A total of 1,280 stool specimens were examined; 158 of them were collected during episodes of diarrhea, and 1,122 were collected during symptom-free periods, every 2 to 3 weeks, from every child. Of the children, 20 (15.4%) excreted Cryptosporidium oocysts during the observation period. Of the diarrheal episodes, 13 (8.3%) were associated with Cryptosporidium sp. Of the control specimens, seven (0.6%) were positive for oocysts. Most of the infections were documented during the months of February to May, at the end of the dry season. Cryptosporidium infections are very common among Guatemalan infants and are an important cause of diarrhea and weight loss. The introduction of liquid or solid foods in the diets of the children, the presence of domestic animals (dogs, cats, or poultry), and the absence of toilet facilities in the house seem to be important risk factors for infection; also, deficient nutritional status may predispose the infected child to Cryptosporidium-associated illness. PMID:3343318

  10. Efficacy of gaseous chlorine dioxide as a sanitizer against Cryptosporidium parvum, Cyclospora cayetanensis, and Encephalitozoon intestinalis on produce.

    PubMed

    Ortega, Ynes R; Mann, Amy; Torres, Maria P; Cama, Vitaliano

    2008-12-01

    The efficacy of gaseous chlorine dioxide to reduce parasite and bacterial burden in produce was studied. Basil and lettuce leaves were inoculated with Cryptosporidium parvum and Cyclospora cayetanensis oocysts, Encephalitozoon intestinalis spores, and a cocktail of two isolates of nalidixic acid-resistant Escherichia coli O157:H7. The inoculated samples were then treated for 20 min with gaseous chlorine dioxide at 4.1 mg/liter. Cryptosporidium had a 2.6 and 3.31 most-probable-number log reduction in basil and lettuce, respectively. Reduction of Encephalitozoon in basil and lettuce was 3.58 and 4.58 CFU/g respectively. E. coli loads were significantly reduced (2.45 to 3.97 log), whereas Cyclospora sporulation was not affected by this treatment. PMID:19244892

  11. Effect of PAC addition on immersed ultrafiltration for the treatment of algal-rich water.

    PubMed

    Zhang, Yan; Tian, Jiayu; Nan, Jun; Gao, ShanShan; Liang, Heng; Wang, Meilian; Li, Guibai

    2011-02-28

    The aim of this study was to evaluate the effect of powdered activated carbon (PAC) addition on the treatment of algal-rich water by immersed ultrafiltration (UF), in terms of permeate quality and membrane fouling. Experiments were performed with a hollow-fiber polyvinyl chloride ultrafiltration membrane at a laboratory scale, 20-25°C and 10 L/(m(2) h) constant permeate flux. UF could achieve an absolute removal of Microcystis aeruginosa cells, but a poor removal of algogenic organic matter (AOM) released into water, contaminants responsible for severe membrane fouling. The addition of 4 g/L PAC to the immersed UF reactor significantly alleviated the development of trans-membrane pressure and enhanced the removal of dissovled organic carbon (by 10.9±1.7%), UV(254) (by 27.1±1.7%), and microcystins (expressed as MC-LR(eq), by 40.8±4.2%). However, PAC had little effect on the rejection of hydrophilic high molecular weight AOM such as carbohydrates and proteins. It was also identified that PAC reduced the concentrations of carbohydrates and proteins in the reactor due to decreased light intensity, as well as the MC-LR(eq) concentration by PAC adsorption. PMID:21216530

  12. Cryptosporidium Prevalence and Risk Factors among Mothers and Infants 0 to 6 Months in Rural and Semi-Rural Northwest Tanzania: A Prospective Cohort Study

    PubMed Central

    Pedersen, Sarah H.; Wilkinson, Amanda L.; Andreasen, Aura; Warhurst, David C.; Kinung'hi, Safari M.; Urassa, Mark; Mkwashapi, Denna M.; Todd, Jim; Changalucha, John; McDermid, Joann M.

    2014-01-01

    ineffective treatment in young infants, minimizing the overall environmental burden (e.g. contaminated water) and particularly, maternal Cryptosporidium infection burden as a means to protect against early infant infection needs prioritization. PMID:25275519

  13. Biochar Addition to Stormwater Treatment Media for Enhanced Removal of Nitrogen

    NASA Astrophysics Data System (ADS)

    Imhoff, P. T.; Jin, J.; Tian, J.; Chiu, P.; Guo, M.

    2015-12-01

    Urban stormwater management systems, such as bioretention facilities, require substantial land area and are often ineffective in removing nitrogen. This project seeks to improve nitrogen removal in bioretention media by modifying the hydraulic and treatment characteristics of the infiltration medium with biochar addition. A commercial wood biochar pyrolyzed from Southern Yellow Pine at 500°C was used. Laboratory experiments demonstrated that biochar addition to a typical bioretention medium (soil-mix: 4% saw dust, 88% sand, 8% clay) increased ammonium sorption at typical stormwater concentrations (2 mg/L) by a factor of 6, total porosity by 16.6%, and water retention at most matric potentials. The effect of the biochar-amended medium on nitrate removal was evaluated in pilot-scale experiments. Side-by-side experimental cells (91 cm dia., 1.2 m deep) were constructed to treat stormwater runoff from a parking lot. The control cell contained 100% soil mix while the biochar cell contained 4% biochar and 96% soil-mix by mass. Treatment media were 76.2 cm in depth and overlain by 5.1 cm of wood mulch in both cells, with a water table maintained at the bottom of the treatment zones. Cells were instrumented with TDR moisture sensors, pressure transducers, and redox and temperature sensors. Two pilot-scale experiments were conducted that included a bromide tracer and nitrate with a hydraulic loading of 5.5cm/h for 24 h in early spring and 36 h in summer. Effluent was continuously sampled for nitrogen compounds during these tests. Tracer tests and TDR measurements showed that biochar increased the average volumetric water content of the vadose zone by 14.7% and the mean residence time by 12.6%. For the spring field test at 14°C, nitrate in the control cell effluent increased by 6.1% but decreased by 43.5% for the biochar cell. For the summer field test at 22°C, 30.6% and 84.7% of influent nitrate was removed in the control and biochar cells, respectively. In the summer

  14. Quantification of viable Giardia cysts and Cryptosporidium oocysts in wastewater using propidium monoazide quantitative real-time PCR.

    PubMed

    Alonso, José L; Amorós, Inmaculada; Guy, Rebecca A

    2014-07-01

    Real-time PCR (qPCR) is a rapid tool to quantify pathogens in the aquatic environment; however, it quantifies all pathogens, including both viable and nonviable. Propidium monoazide (PMA) is a membrane-impairment dye that penetrates only membrane-damaged cells. Once inside the cell, PMA is covalently cross-linked to DNA through light photoactivation, and PCR amplification is strongly inhibited. The goal of this study was to evaluate PMA-qPCR assays for rapid quantification of viable and heat-treated Giardia cysts and Cryptosporidium oocysts in wastewater. We observed a reduction in detection of heat-treated Giardia duodenalis cysts of 83.2, 89.9, 98.2, or 97% with PMA-qPCR assays amplifying a 75 base-pair (bp) β-giardin target, 77-bp triosephosphate isomerase (tpi), 133-bp glutamate dehydrogenase (GDH), and 143-bp β-giardin gene target, respectively. Thus, the exclusion of dead cysts was more effective when qPCR assays that produced larger amplicons were used. The PMA treatment of Cryptosporidium oocysts plus/minus heat treatment abolished the fluorescent signal for dead oocysts with a PMA-qPCR assay amplifying a Cryptosporidium parvum (150-bp) oocyst wall protein (COWP) gene. The PMA-qPCR 143-bp β-giardin assay for Giardia and the PMA-qPCR 150-bp COWP assay for Cryptosporidium accurately quantified live oo(cysts), and failed to detect dead oo(cysts), when phosphate-buffered saline and tertiary effluent wastewater were spiked with concentrations of 10(3) or 10(2) dead oo(cysts), respectively. Therefore, these assays are suitable for the detection of viable parasites that are typically present in tertiary wastewater effluents at concentrations of <10(3) oo(cysts)/l and can provide rapid risk assessments of environmental water. PMID:24781028

  15. Effect of tillage and rainfall on transport of manure-applied Cryptosporidium parvum oocysts through soil.

    PubMed

    Ramirez, Norma E; Wang, Ping; Lejeune, Jeff; Shipitalo, Martin J; Ward, Lucy A; Sreevatsan, Srinand; Dick, Warren A

    2009-01-01

    Most waterborne outbreaks of cryptosporidiosis have been attributed to agricultural sources due to the high prevalence of Cryptosporidium oocysts in animal wastes and manure spreading on farmlands. No-till, an effective conservation practice, often results in soil having higher water infiltration and percolation rates than conventional tillage. We treated six undisturbed no-till and six tilled soil blocks (30 by 30 by 30 cm) with 1 L liquid dairy manure containing 10(5) C. parvum oocysts per milliliter to test the effect of tillage and rainfall on oocyst transport. The blocks were subjected to rainfall treatments consisting of 5 mm or 30 mm in 30 min. Leachate was collected from the base of the blocks in 35-mL increments using a 64-cell grid lysimeter. Even before any rain was applied, approximately 300 mL of water from the liquid manure (30% of that applied) was transported through the no-till soil, but none through the tilled blocks. After rain was applied, a greater number and percentage of first leachate samples from the no-till soil blocks compared to the tilled blocks tested positive for Cryptosporidium oocysts. In contrast to leachate, greater numbers of oocysts were recovered from the tilled soil, itself, than from the no-till soil. Although tillage was the most important factor affecting oocyst transport, rainfall timing and intensity were also important. To minimize transport of Cryptosporidium in no-till fields, manure should be applied at least 48 h before heavy rainfall is anticipated or methods of disrupting the direct linkage of surface soil to drains, via macropores, need to be used. PMID:19875795

  16. Short report: possible Cryptosporidium muris infection in humans.

    PubMed

    Katsumata, T; Hosea, D; Ranuh, I G; Uga, S; Yanagi, T; Kohno, S

    2000-01-01

    Oocysts of cryptosporidia whose morphology resembled that of Cryptosporidium muris were found in the stool of 2 healthy girls in Surabaya, Indonesia. The oocysts were predominantly oval and measured 7.75+/-0.17 x 5.55+/-0.13 microm (mean+/-SD). The number of oocysts excreted were more than 10(5) per gram of stool. The oocysts were well stained with fluorescein-conjugated monoclonal antibody to Cryptosporidium. The specimens from both girls containing the oocysts showed a positive result by the polymerase chain reaction (PCR) using primers specific for the genus Cryptosporidium, but a negative result by the PCR using primers specific for C. parvum. The 2 girls passed oocysts for 5 and 6 days, respectively. They did not complain of any symptoms during the passage of oocysts. PMID:10761726

  17. Cryptosporidium Species and Subtypes and Clinical Manifestations in Children, Peru

    PubMed Central

    Cama, Vitaliano A.; Bern, Caryn; Roberts, Jacqueline; Cabrera, Lilia; Sterling, Charles R.; Ortega, Ynes; Gilman, Robert H.

    2008-01-01

    To determine whether clinical manifestations are associated with genotypes or subtypes of Cryptosporidium spp., we studied a 4-year longitudinal birth cohort of 533 children in Peru. A total of 156 infection episodes were found in 109 children. Data from first infections showed that C. hominis was associated with diarrhea, nausea, vomiting, general malaise, and increased oocyst shedding intensity and duration. In contrast, C. parvum, C. meleagridis, C. canis, and C. felis were associated with diarrhea only. C. hominis subtype families were identified (Ia, Ib, Id, and Ie); all were associated with diarrhea. Ib was also associated with nausea, vomiting, and general malaise. All C. parvum specimens belonged to subtype family IIc. Analysis of risk factors did not show associations with specific Cryptosporidium spp. genotypes or subtypes. These findings strongly suggest that Cryptosporidium spp. and subtypes are linked to different clinical manifestations in children. PMID:18826821

  18. Detection of a novel genotype of Cryptosporidium in Antarctic pinnipeds.

    PubMed

    Rengifo-Herrera, Claudia; Ortega-Mora, Luis Miguel; Gómez-Bautista, Mercedes; García-Peña, Francisco Javier; García-Párraga, Daniel; Pedraza-Díaz, Susana

    2013-01-16

    A study was conducted to investigate the presence of Cryptosporidium and Giardia in Antarctic marine mammals. A total of 270 faecal samples from different species of pinnipeds from different locations in the South Shetland Islands and Antarctic Peninsula were analysed by immunofluorescence microscopy and PCR. Cryptosporidium was detected by PCR in three samples from Southern elephant seals (Mirounga leonina) and 2 Weddell seals (Leptonychotes weddellii). However, no oocysts were observed in any of the samples by immunofluorescence microscopy. Molecular characterisation of the isolates, using the 18S rDNA, the HSP70 and the COWP loci, revealed the presence of a Cryptosporidium sp., previously reported from an Antarctic Southern elephant seal, in the elephant seals and a novel genotype in Weddell seals. Giardia could not be detected in any of the samples analysed. PMID:23021408

  19. Effect of cyanuric acid on the inactivation of Cryptosporidium parvum under hyperchlorination conditions.

    PubMed

    Murphy, Jennifer L; Arrowood, Michael J; Lu, Xin; Hlavsa, Michele C; Beach, Michael J; Hill, Vincent R

    2015-06-16

    Cyanuric acid (CYA) is a chlorine stabilizer used in swimming pools to limit UV degradation of chlorine, thus reducing chlorine use and cost. However, CYA has been shown to decrease the efficacy of chlorine disinfection. In the event of a diarrheal incident, CDC recommends implementing 3-log10 inactivation conditions for Cryptosporidium (CT value = 15 300 mg·min/L) to remediate pools. Currently, CYA's impact on Cryptosporidium inactivation is not fully determined. We investigated the impact of multiple concentrations of CYA on C. parvum inactivation (at 20 and 40 mg/L free chlorine; average pH 7.6; 25 °C). At 20 mg/L free chlorine, average estimated 3-log10 CT values were 17 800 and 31 500 mg·min/L with 8 and 16 mg/L CYA, respectively, and the average estimated 1-log10 CT value was 76 500 mg·min/L with 48 mg/L CYA. At 40 mg/L free chlorine, 3-log10 CT values were lower than those at 20 mg/L, but still higher than those of free chlorine-only controls. In the presence of ∼100 mg/L CYA, average 0.8- and 1.4-log10 reductions were achieved by 72 h at 20 and 40 mg/L free chlorine, respectively. This study demonstrates CYA significantly delays chlorine inactivation of Cryptosporidium oocysts, emphasizing the need for additional pool remediation options following fecal incidents. PMID:26042636

  20. Coevolution of Cryptosporidium tyzzeri and the house mouse (Mus musculus)✯

    PubMed Central

    Kváč, Martin; McEvoy, John; Loudová, Martina; Stenger, Brianna; Sak, Bohumil; Květoňová, Dana; Ditrich, Oleg; Rašková, Veronika; Moriarty, Elaine; Rost, Michael; Macholán, Miloš; Piálek, Jaroslav

    2013-01-01

    Two house mouse subspecies occur in Europe, eastern and northern Mus musculus musculus (Mmm) and western and southern Mus musculus domesticus (Mmd). A secondary hybrid zone occurs where their ranges meet, running from Scandinavia to the Black Sea. In this paper, we tested a hypothesis that the apicomplexan protozoan species Cryptosporidium tyzzeri has coevolved with the house mouse. More specifically, we assessed to what extent the evolution of this parasite mirrors divergence of the two subspecies. In order to test this hypothesis, we analyzed sequence variation at five genes (ssrRNA, Cryptosporidium oocyst wall protein (COWP), thrombospondin-related adhesive protein of Cryptosporidium 1 (TRAP-C1), actin and gp60) in C. tyzzeri isolates from Mmd and Mmm sampled along a transect across the hybrid zone from the Czech Republic to Germany. Mmd samples were supplemented with mice from New Zealand. We found two distinct isolates of C. tyzzeri, each occurring exclusively in one of the mouse subspecies (C. tyzzeri-Mmm and C. tyzzeri-Mmd). In addition to genetic differentiation, oocysts of the C. tyzzeri-Mmd subtype (mean: 4.24 × 3.69 μm) were significantly smaller than oocysts of C. tyzzeri-Mmm (mean: 4.49 × 3.90 μm). Mmm and Mmd were susceptible to experimental infection with both C. tyzzeri subtypes; however, the subtypes were not infective for the rodent species Meriones unguiculatus, Mastomys coucha, Apodemus flavicollis or Cavia porcellus. Overall, our results support the hypothesis that C. tyzzeri is coevolving with Mmm and Mmd. PMID:23791796

  1. Can Additional Homeopathic Treatment Save Costs? A Retrospective Cost-Analysis Based on 44500 Insured Persons

    PubMed Central

    Ostermann, Julia K.; Reinhold, Thomas; Witt, Claudia M.

    2015-01-01

    Objectives The aim of this study was to compare the health care costs for patients using additional homeopathic treatment (homeopathy group) with the costs for those receiving usual care (control group). Methods Cost data provided by a large German statutory health insurance company were retrospectively analysed from the societal perspective (primary outcome) and from the statutory health insurance perspective. Patients in both groups were matched using a propensity score matching procedure based on socio-demographic variables as well as costs, number of hospital stays and sick leave days in the previous 12 months. Total cumulative costs over 18 months were compared between the groups with an analysis of covariance (adjusted for baseline costs) across diagnoses and for six specific diagnoses (depression, migraine, allergic rhinitis, asthma, atopic dermatitis, and headache). Results Data from 44,550 patients (67.3% females) were available for analysis. From the societal perspective, total costs after 18 months were higher in the homeopathy group (adj. mean: EUR 7,207.72 [95% CI 7,001.14–7,414.29]) than in the control group (EUR 5,857.56 [5,650.98–6,064.13]; p<0.0001) with the largest differences between groups for productivity loss (homeopathy EUR 3,698.00 [3,586.48–3,809.53] vs. control EUR 3,092.84 [2,981.31–3,204.37]) and outpatient care costs (homeopathy EUR 1,088.25 [1,073.90–1,102.59] vs. control EUR 867.87 [853.52–882.21]). Group differences decreased over time. For all diagnoses, costs were higher in the homeopathy group than in the control group, although this difference was not always statistically significant. Conclusion Compared with usual care, additional homeopathic treatment was associated with significantly higher costs. These analyses did not confirm previously observed cost savings resulting from the use of homeopathy in the health care system. PMID:26230412

  2. An additional simple denitrification bioreactor using packed gel envelopes applicable to industrial wastewater treatment.

    PubMed

    Morita, Masahiko; Uemoto, Hiroaki; Watanabe, Atsushi

    2007-08-15

    A simple denitrification bioreactor for nitrate-containing wastewater without organic compounds was developed. This bioreactor consisted of packed gel envelopes in a single tank. Each envelope comprised two plates of gels containing Paracoccus denitrificans cells with an internal space between the plates. As an electron donor for denitrification, ethanol was injected into the internal space and not directly into the wastewater. P. denitrificans cells in the gel reduced nitrate to nitrogen gas by using the injected ethanol. Nitrate-containing desulfurization wastewater derived from a coal-fired thermal power plant was continuously treated with 20 packed gel envelopes (size, 1,000 x 900 x 12 mm; surface area, 1.44 m(2)) in a reactor tank (volume 1.5 m(3)). When the total nitrogen concentration in the inflow was around 150 mg-N x L(-1), the envelopes removed approximately 60-80% of the total nitrogen, and the maximum nitrogen removal rate was 5.0 g-N x day(-1) per square meter of the gel surface. This value corresponded to the volumetric nitrogen removal performance of 0.109 kg-N x m(-3) x day(-1). In each envelope, a high utilization efficiency of the electron donor was attained, although more than the double amount of the electron donor was empirically injected in the present activated sludge system to achieve denitrification when compared with the theoretical value. The bioreactor using the envelopes would be extremely effective as an additional denitrification system because these envelopes can be easily installed in the vacant spaces of preinstalled water treatment systems, without requiring additional facilities for removing surplus ethanol and sludge. PMID:17252606

  3. Long-Term Transport of Cryptosporidium Parvum

    NASA Astrophysics Data System (ADS)

    Andrea, C.; Harter, T.; Hou, L.; Atwill, E. R.; Packman, A.; Woodrow-Mumford, K.; Maldonado, S.

    2005-12-01

    The protozoan pathogen Cryptosporidium parvum is a leading cause of waterborne disease. Subsurface transport and filtration in natural and artificial porous media are important components of the environmental pathway of this pathogen. It has been shown that the oocysts of C. parvum show distinct colloidal properties. We conducted a series of laboratory studies on sand columns (column length: 10 cm - 60 cm, flow rates: 0.7 m/d - 30 m/d, ionic strength: 0.01 - 100 mM, filter grain size: 0.2 - 2 mm, various solution chemistry). Breakthrough curves were measured over relatively long time-periods (hundreds to thousands of pore volumes). We show that classic colloid filtration theory is a reasonable tool for predicting the initial breakthrough, but it is inadequate to explain the significant tailing observed in the breakthrough of C. parvum oocyst through sand columns. We discuss the application of the Continuous Time Random Walk approach to account for the strong tailing that was observed in our experiments. The CTRW is generalized transport modeling framework, which includes the classic advection-dispersion equation (ADE), the fractional ADE, and the multi-rate mass transfer model as special cases. Within this conceptual framework, it is possible to distinguish between the contributions of pore-scale geometrical (physical) disorder and of pore-scale physico-chemical heterogeneities (e.g., of the filtration, sorption, desorption processes) to the transport of C. parvum oocysts.

  4. Cheap and Cheerful Stream Restoration - An Example of System Wide Woody Addition Treatment

    NASA Astrophysics Data System (ADS)

    Wheaton, J. M.; Bennett, S. N.; Bouwes, N.; Camp, R.

    2012-12-01

    Stream restoration has been plagued with high price tags, limited spatial extents, and questionable effectiveness in light of largely absent monitoring efforts. One prominent example is the placement of large woody debris (LWD) structures and engineered log jams that are frequently employed to promote heterogeneity of instream habitat. Ironically, many of these treatments attempt to lock in place and over-engineer the woody structures as opposed to allowing them to adjust and rearrange themselves as natural LWD would have. We are in the midst of a large scale restoration experiment using LWD to recover ESA-listed steelhead trout (Oncorhynchus mykiss) populations in the Asotin Creek Watershed of Southeast Washington. The project is an Intensively Monitored Watershed (IMW) where the restoration treatment and monitoring use a hierarchal staircase design maximizing the power to detect a population level response in steelhead. We are treating over 12 km of stream with enough LWD input (> 200 pieces per km) to mimic the historic background wood loading and encourage the stream to reshape and regularly rework itself leaving. We are using hundreds of structures we call DWS (dynamic woody structures), which generally consist of a series of wooden fence posts driven into the stream bed and complex LWD anchored between them to invoke a specific hydrogeomorphic response. The real advantage of these DWS are their cost. They can be installed quickly (15-30 minutes each) and cheaply (< $100/DWS); even in remote settings with a 2-3 person crew, hydraulic post pounder, very cheap materials, and avoiding impacts associated with operating heavy equipment. This allows us to install lots of the structures at high density (every 5-15 channel widths) over an entire stream system. We call this overall approach System Wide Woody Addition Treatment (SWWAT). In the long term, we hypothesize that the SWWAT will provide an intial input LWD that will become a part of study creeks which are more

  5. Influence of attrition scrubbing, ultrasonic treatment, and oxidant additions on uranium removal from contaminated soils

    SciTech Connect

    Timpson, M.E.; Elless, M.P.; Francis, C.W.

    1994-06-01

    As part of the Uranium in Soils Integrated Demonstration Project being conducted by the US Department of Energy, bench-scale investigations of selective leaching of uranium from soils at the Fernald Environmental Management Project site in Ohio were conducted at Oak Ridge National Laboratory. Two soils (storage pad soil and incinerator soil), representing the major contaminant sources at the site, were extracted using carbonate- and citric acid-based lixiviants. Physical and chemical processes were used in combination with the two extractants to increase the rate of uranium release from these soils. Attrition scrubbing and ultrasonic dispersion were the two physical processes utilized. Potassium permanganate was used as an oxidizing agent to transform tetravalent uranium to the hexavalent state. Hexavalent uranium is easily complexed in solution by the carbonate radical. Attrition scrubbing increased the rate of uranium release from both soils when compared with rotary shaking. At equivalent extraction times and solids loadings, however, attrition scrubbing proved effective only on the incinerator soil. Ultrasonic treatments on the incinerator soil removed 71% of the uranium contamination in a single extraction. Multiple extractions of the same sample removed up to 90% of the uranium. Additions of potassium permanganate to the carbonate extractant resulted in significant changes in the extractability of uranium from the incinerator soil but had no effect on the storage pad soil.

  6. A fast method for detecting Cryptosporidium parvum oocysts in real world samples

    NASA Astrophysics Data System (ADS)

    Stewart, Shona; McClelland, Lindy; Maier, John

    2005-04-01

    Contamination of drinking water with pathogenic microorganisms such as Cryptosporidium has become an increasing concern in recent years. Cryptosporidium oocysts are particularly problematic, as infections caused by this organism can be life threatening in immunocompromised patients. Current methods for monitoring and analyzing water are often laborious and require experts to conduct. In addition, many of the techniques require very specific reagents to be employed. These factors add considerable cost and time to the analytical process. Raman spectroscopy provides specific molecular information on samples, and offers advantages of speed, sensitivity and low cost over current methods of water monitoring. Raman spectroscopy is an optical method that has demonstrated the capability to identify and differentiate microorganisms at the species and strain levels. In addition, this technique has exhibited sensitivities down to the single organism detection limit. We have employed Raman spectroscopy and Raman Chemical Imaging, in conjunction with chemometric techniques, to detect small numbers of oocysts in the presence of interferents derived from real-world water samples. Our investigations have also indicated that Raman Chemical Imaging may provide chemical and physiological information about an oocyst sample which complements information provided by the traditional methods. This work provides evidence that Raman imaging is a useful technique for consideration in the water quality industry.

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

    PubMed

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

    2005-03-01

    We compared serological responses to Cryptosporidium parvum antigens using surplus sera from females undergoing routine screening for pregnancy from three counties in Hungary where bank-filtered surface water, conventionally filtered and disinfected surface water, and groundwater from either a karst or confined aquifer are commonly used for drinking water. The primary purpose was to determine whether the prevalence and intensity of serological responses, indicators of prior Cryptosporidium infection were similar for these populations. Women using groundwater from a confined aquifer had significantly lower mean serological responses for both the 15/17-kDa and 27-kDa (p < 0.0001) antigen groups than women using conventionally filtered and disinfected surface water or karst well water. This is suggestive of less frequent infections. Women using bank-filtered water also had lower mean responses for both antigen groups. Among women using bank-filtered water, the mean intensity of response for both antigen groups was almost one-third of the mean response observed for women using conventionally filtered and disinfected surface water. These findings suggest that riverbank filtration may be an effective alternative to conventional treatment for reducing Cryptosporidium exposures and infection from surface drinking water sources. PMID:15952455

  8. Quantitative microbial risk assessment of Cryptosporidium and Giardia in well water from a native community of Mexico.

    PubMed

    Balderrama-Carmona, Ana Paola; Gortáres-Moroyoqui, Pablo; Álvarez-Valencia, Luis Humberto; Castro-Espinoza, Luciano; Balderas-Cortés, José de Jesús; Mondaca-Fernández, Iram; Chaidez-Quiroz, Cristóbal; Meza-Montenegro, María Mercedes

    2015-01-01

    Cryptosporidium and Giardia are gastrointestinal disease-causing organisms transmitted by the fecal-oral route, zoonotic and prevalent in all socioeconomic segments with greater emphasis in rural communities. The goal of this study was to assess the risk of cryptosporidiosis and giardiasis of Potam dwellers consuming drinking water from communal well water. To achieve the goal, quantitative microbial risk assessment (QMRA) was carried out as follows: (a) identification of Cryptosporidium oocysts and Giardia cysts in well water samples by information collection rule method, (b) assessment of exposure to healthy Potam residents, (c) dose-response modelling, and (d) risk characterization using an exponential model. All well water samples tested were positive for Cryptosporidium and Giardia. The QMRA results indicate a mean of annual risks of 99:100 (0.99) for cryptosporidiosis and 1:1 (1.0) for giardiasis. The outcome of the present study may drive decision-makers to establish an educational and treatment program to reduce the incidence of parasite-borne intestinal infection in the Potam community, and to conduct risk analysis programs in other similar rural communities in Mexico. PMID:25494486

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

    EPA Science Inventory

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

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

    PubMed

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

    2015-05-15

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

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

    PubMed Central

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

    2015-01-01

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

  12. Combined cetuximab and genistein treatment shows additive anti-cancer effect on oral squamous cell carcinoma.

    PubMed

    Park, Sung-Jin; Kim, Myung-Jin; Kim, Yu-Kyoung; Kim, Soung-Min; Park, Ju-Yong; Myoung, Hoon

    2010-06-01

    The purpose of this study was to evaluate the potency of EGFR pathway inhibition achieved by combining cetuximab, an anti-EGFR monoclonal antibody, and genistein, a tyrosine kinase inhibitor, which target extracellular and intracellular domains of the receptor, respectively, in oral squamous cell carcinoma (OSCC) in vitro and in vivo. Two OSCC cell lines, HSC3 and KB, were treated with cetuximab (C, 0-400mug/ml), genistein (G, 0-80muM), or a combination of both at a range of concentrations. Downstream protein expression of EGFR, p-EGFR, and p-Akt were evaluated by Western blot. Cell proliferation and apoptosis indices were calculated to assess anti-cancer effects in vitro. The in vivo effects of cetuximab and genistein on tumor cell growth were examined using an OSCC xenografted nude mouse model and immunohistochemical analyses of proliferation (PCNA) and microvessel density (CD31). Treatment of cells with dual anti-EGFR agents reduced the expressions of p-EGFR, and p-Akt in HSC3 cell line, but there was no significant difference in downregulation between cetuximab alone and in combination with genistein in KB cells. Both HSC3 and KB cells showed a dose-dependent decrease in cell proliferation significantly with single agent treatment and combination (p<0.05). In low concentration, combined cetuximab and genistein therapy resulted in additive growth inhibition and more apoptosis compared to that achieved with single-agent exposure in both cell lines. A combination of cetuximab and genistein significantly inhibited tumor growth and caused a substantial growth delay in in vivo models of both cell lines while each single-agent exposure caused no delay of tumor growth. Immunohistochemical staining with PCNA revealed that the group receiving combined cetuximab and genistein exhibited the lowest number of proliferating cells and microvessel density (p<0.05). Combined therapy with genistein and cetuximab can add the potency of EGFR signaling inhibition. Because not all

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

    PubMed

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

    2013-02-01

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

  14. North American tree squirrels and ground squirrels with overlapping ranges host different Cryptosporidium species and genotypes.

    PubMed

    Stenger, Brianna L S; Clark, Mark E; Kváč, Martin; Khan, Eakalak; Giddings, Catherine W; Prediger, Jitka; McEvoy, John M

    2015-12-01

    Wildlife-associated Cryptosporidium are an emerging cause of cryptosporidiosis in humans. The present study was undertaken to determine the extent to which North American tree squirrels and ground squirrels host zoonotic Cryptosporidium species and genotypes. Fragments of the Cryptosporidium 18S rRNA and actin genes were amplified and sequenced from fecal samples obtained from three tree squirrel and three ground squirrel species. In tree squirrels, Cryptosporidium was identified in 40.5% (17/42) of American red squirrels (Tamiasciurus hudsonicus), 40.4% (55/136) of eastern gray squirrels (Sciurus carolinensis), and 28.6% (2/7) of fox squirrels (Sciurus niger). Human-pathogenic Cryptosporidium ubiquitum and Cryptosporidium skunk genotype were the most prevalent species/genotypes in tree squirrels. Because tree squirrels live in close proximity to humans and are frequently infected with potentially zoonotic Cryptosporidium species/genotypes, they may be a significant reservoir of infection in humans. In ground squirrels, Cryptosporidium was detected in 70.2% (33/47) of 13-lined ground squirrels (Ictidomys tridecemlineatus), 35.1% (27/77) of black-tailed prairie dogs (Cynomys ludovicianus), and the only golden-mantled ground squirrel (Callospermophilus lateralis) that was sampled. Cryptosporidium rubeyi and ground squirrel genotypes I, II, and III were identified in isolates from these ground squirrel species. In contrast to the Cryptosporidium infecting tree squirrels, these species/genotypes appear to be specific for ground squirrels and are not associated with human disease. PMID:26437239

  15. Efficacy of a potential DNA vaccine encoding Cryptosporidium baileyi rhomboid protein against homologous challenge in chickens.

    PubMed

    Yang, Yimin; Xue, Xue; Yang, Yi; Chen, Xueqiu; Du, Aifang

    2016-07-30

    The parasite Cryptosporidium baileyi can infect the larynx, trachea, bursa and cloaca of poultry, causing high mortality during severe infection and leading to substantial economic losses of the poultry industry. The rhomboid protein is very important in Cryptosporidium infection. In this study, a nucleic acid based vaccine candidate pEGFP-CbROM was constructed. After orally challenging with C. baileyi oocysts, the corresponding immune responses induced were analyzed and the immunoprotective effect evaluated in chickens. Obtained results revealed that this nucleic acid based vaccine could induce antibody responses and peripheral blood T lymphocytes proliferation significantly (P<0.05), while the peripheral blood B lymphocyte proliferation increased significantly (P<0.05) only at a high dose of 100μg of pEGFP-CbROM, compared with the PBS control group. After C. baileyi infection, the duration of oocysts shedding was shortened by 2days in the 100μg pEGFP-CbROM group, and the rate of reduction could reach to around 71.3%. While no significant difference in body weight gain was observed among the immunized groups (P>0.05), the differences between the immunized and the non-immunized groups were found to be significant (P<0.05). Our data provides a useful basis for further work in cryptosporidiosis prevention and treatment. PMID:27369569

  16. Substituted Pyrrolo[2,3-d]pyrimidines as Cryptosporidium hominis Thymidylate Synthase Inhibitors

    PubMed Central

    Kumar, Vidya P.; Frey, Kathleen M.; Wang, Yiqiang; Jain, Hitesh K.; Gangjee, Aleem; Anderson, Karen S.

    2013-01-01

    Cryptosporidiosis, a gastrointestinal disease caused by a protozoan Cryptosporidium hominis is often fatal in immunocompromised individuals. There is little clinical data to show that the existing treatment by nitazoxanide and paromomycin is effective in immunocompromised individuals1, 2. Thymidylate synthase (TS) and dihydrofolate reductase (DHFR) are essential enzymes in the folate biosynthesis pathway and are well established as drug targets in cancer and malaria. A novel series of classical antifolates, 2-amino-4-oxo-5-substituted pyrrolo[2,3-d]pyrimidines have been evaluated as Cryptosporidium hominis thymidylate synthase (ChTS) inhibitors. Crystal structure in complex with the most potent compound, a 2’-chlorophenyl with a sulfur bridge with a Ki of 8.83 ± 0.67 nM is discussed in terms of several Van de Waals, hydrophobic and hydrogen bond interactions with the protein residues and the substrate analog 5-fluorodeoxyuridine monophosphate. Of these interactions, two interactions with the non-conserved residues (A287 and S290) offer an opportunity to develop ChTS specific inhibitors. Compound 6 serves as a lead compound for analog design and its crystal structure provides clues for the design of ChTS specific inhibitors. PMID:23927969

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

    PubMed

    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. PMID:11317885

  18. Selective and potent urea inhibitors of Cryptosporidium parvum inosine 5′ monophosphate dehydrogenase

    PubMed Central

    Gorla, Suresh Kumar; Kavitha, Mandapati; Zhang, Minjia; Liu, Xiaoping; Sharling, Lisa; Gollapalli, Deviprasad R.; Striepen, Boris; Hedstrom, Lizbeth; Cuny, Gregory D.

    2012-01-01

    Cryptosporidium parvum and related species are zoonotic intracellular parasites of the intestine. Cryptosporidium is a leading cause of diarrhea in small children around the world. Infection can cause severe pathology in children and immunocompromised patients. This waterborne parasite is resistant to common methods of water treatment and therefore a prominent threat to drinking and recreation water even in countries with strong water safety systems. The drugs currently used to combat these organisms are ineffective. Genomic analysis revealed that the parasite relies solely on inosine-5′-monophosphate dehydrogenase (IMPDH) for the biosynthesis of guanine nucleotides. Herein, we report a selective urea-based inhibitor of C. parvum IMPDH (CpIMPDH) identified by high throughput screening. We performed a SAR study of these inhibitors with some analogues exhibiting high potency (IC50 < 2 nM) against CpIMPDH, excellent selectivity > 1000-fold versus human IMPDH type 2 and good stability in mouse liver microsomes. A subset of inhibitors also displayed potent antiparasitic activity in a Toxoplasma gondii model. PMID:22950983

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

    PubMed Central

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

    2012-01-01

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

  20. A modified EPA Method 1623 that uses tangential flow hollow-fiber ultrafiltration and heat dissociation steps to detect waterborne Cryptosporidium and Giardia spp.

    PubMed

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

    2012-01-01

    Cryptosporidium and Giardia species are two of the most prevalent protozoa that cause waterborne diarrheal disease outbreaks worldwide. To better characterize the prevalence of these pathogens, EPA Method 1623 was developed and used to monitor levels of these organisms in US drinking water supplies (12). The method has three main parts; the first is the sample concentration in which at least 10 L of raw surface water is filtered. The organisms and trapped debris are then eluted from the filter and centrifuged to further concentrate the sample. The second part of the method uses an immunomagnetic separation procedure where the concentrated water sample is applied to immunomagnetic beads that specifically bind to the Cryptosporidium oocysts and Giardia cysts allowing for specific removal of the parasites from the concentrated debris. These (oo)cysts are then detached from the magnetic beads by an acid dissociation procedure. The final part of the method is the immunofluorescence staining and enumeration where (oo)cysts are applied to a slide, stained, and enumerated by microscopy. Method 1623 has four listed sample concentration systems to capture Cryptosporidium oocysts and Giardia cysts in water: Envirochek filters (Pall Corporation, Ann Arbor, MI), Envirochek HV filters (Pall Corporation), Filta-Max filters (IDEXX, Westbrook, MA), or Continuous Flow Centrifugation (Haemonetics, Braintree, MA). However, Cryptosporidium and Giardia (oo)cyst recoveries have varied greatly depending on the source water matrix and filters used(1,14). A new tangential flow hollow-fiber ultrafiltration (HFUF) system has recently been shown to be more efficient and more robust at recovering Cryptosporidium oocysts and Giardia cysts from various water matrices; moreover, it is less expensive than other capsule filter options and can concentrate multiple pathogens simultaneously(1-3,5-8,10,11). In addition, previous studies by Hill and colleagues demonstrated that the HFUF significantly

  1. Genetic modification of the diarrhoeal pathogen Cryptosporidium parvum.

    PubMed

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

    2015-07-23

    Recent studies into the global causes of severe diarrhoea in young children have identified the protozoan parasite Cryptosporidium as the second most important diarrhoeal pathogen after rotavirus. Diarrhoeal disease is estimated to be responsible for 10.5% of overall child mortality. Cryptosporidium is also an opportunistic pathogen in the contexts of human immunodeficiency virus (HIV)-caused AIDS and organ transplantation. There is no vaccine and only a single approved drug that provides no benefit for those in gravest danger: malnourished children and immunocompromised patients. Cryptosporidiosis drug and vaccine development is limited by the poor tractability of the parasite, which includes a lack of systems for continuous culture, facile animal models, and molecular genetic tools. Here we describe an experimental framework to genetically modify this important human pathogen. We established and optimized transfection of C. parvum sporozoites in tissue culture. To isolate stable transgenics we developed a mouse model that delivers sporozoites directly into the intestine, a Cryptosporidium clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system, and in vivo selection for aminoglycoside resistance. We derived reporter parasites suitable for in vitro and in vivo drug screening, and we evaluated the basis of drug susceptibility by gene knockout. We anticipate that the ability to genetically engineer this parasite will be transformative for Cryptosporidium research. Genetic reporters will provide quantitative correlates for disease, cure and protection, and the role of parasite genes in these processes is now open to rigorous investigation. PMID:26176919

  2. Genetic modification of the diarrheal pathogen Cryptosporidium parvum

    PubMed Central

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

    2015-01-01

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

  3. Molecular epidemiology of Cryptosporidium and Giardia in cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryptosporidium spp. and Giardia duodenalis are enteric protozoan parasites that infect a wide range of vertebrate hosts including humans. Infections with both parasites are known as one of the most common causes of diarrhea in humans and livestock. The epidemiology of cryptosporidiosis and giardias...

  4. Identification of Cryptosporidium from Dairy Cattle in Pahang, Malaysia

    PubMed Central

    Hisamuddin, Nur Hazirah; Hashim, Najat; Soffian, Sharmeen Nellisa; Amin, Mohd Hishammfariz Mohd; Wahab, Ridhwan Abdul; Mohammad, Mardhiah; Isa, Muhammad Lokman Md; Yusof, Afzan Mat

    2016-01-01

    Cryptosporidium, a protozoan parasite, can cause cryptosporidiosis which is a gastrointestinal disease that can infect humans and livestock. Cattle are the most common livestock that can be infected with this protozoan. This study was carried out to determine the prevalence of Cryptosporidium infection in cattle in Kuantan, Pahang, Malaysia and to find out the association between the occurrence of infection and 3 different ages of cattle (calves less than 1 year, yearling, and adult cattle). The samples were processed by using formol-ether concentration technique and stained by modified Ziehl Neelsen. The results showed that 15.9% (24/151) of cattle were positive for Cryptosporidium oocysts. The occurrence of Cryptosporidium in calves less than 1 year was the highest with the percentage of 20.0% (11/55) followed by yearling and adult cattle, with the percentage occurrence of 15.6 % (7/45) and 11.8% (6/51), respectively. There was no significant association between the occurrence and age of cattle and presence of diarrhea. Good management practices and proper hygiene management must be taken in order to reduce the infection. It is highly important to control the infection since infected cattle may serve as potential reservoirs of the infection to other animals and humans, especially animal handlers. PMID:27180579

  5. A COMPARISON OF ENUMERATION TECHNIQUES FOR CRYPTOSPORIDIUM PARVUM OOCYSTS

    EPA Science Inventory

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

  6. A highly divergent 33 kDa Cryptosporidium parvum antigen

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  7. DETECTION OF CRYPTOSPORIDIUM OOCYSTS IN SOURCE AND FINISHED WATERS

    EPA Science Inventory

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

  8. THE INFECTIVITY OF CRYPTOSPORIDIUM PARVUM IN HEALTHY VOLUNTEERS

    EPA Science Inventory

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

  9. CRYPTOSPORIDIUM VIRULENCE DETERMINANTS-ARE WE THERE YET? (R828035)

    EPA Science Inventory

    Exposure to Cryptosporidium parvum in healthy individuals results in transient infection that may be asymptomatic or can result in self-limited diarrhoea. In contrast, acquired immune deficiency syndrome patients with cryptosporidiosis can experience severe manifestatio...

  10. Genotyping Cryptosporidium andersoni in Cattle in Shaanxi Province, Northwestern China

    PubMed Central

    Gao, Man; Bian, Qing-Qing; Hu, Bing; Cong, Mei-Mei; Lin, Qing; Wang, Rong-Jun; Qi, Meng; Qi, Mao-Zhen; Zhu, Xing-Quan; Zhang, Long-Xian

    2013-01-01

    The present study examined the prevalence and genotypes of Cryptosporidium andersoni in cattle in Shaanxi province, China. A total of 2071 fecal samples (847 from Qinchuan cattle and 1224 from dairy cattle) were examined for the presence of Cryptosporidium oocysts, and 70 samples (3.4%) were C. andersoni-positive and those positive samples were identified by PCR amplification of the small subunit ribosomal RNA (SSU rRNA) and the Cryptosporidium oocyst wall protein (COWP) genes. C. andersoni was the only species found in the examined cattle in this province. Fifty-seven C. andersoni isolates were characterized into 5 MLST subtypes using multilocus sequence typing analysis, including a new subtype in the native beef breed Qinchuan cattle. All of these C. andersoni isolates presented a clonal genetic structure. These findings provide new insights into the genetic structure of C. andersoni isolates in Shaanxi province and basic data of Cryptosporidium prevalence status, which in turn have implications for controlling cryptosporidiosis in this province. PMID:23560072

  11. Gene expression during excystation of Cryptosporidium parvum oocysts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The present study describes transcription of mRNA from genes encoding metabolic or structural proteins during excystation of Cryptosporidium parvum oocysts. RNA was harvested from C. parvum oocysts before excystation, and at 5, 10, and 15 min during excystation. Subtractive cDNA libraries were pre...

  12. Prevalence of Cryptosporidium in small ruminants from Veracruz, Mexico

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryptosporidiosis is a zoonotic disease caused by the protozoan parasite Cryptosporidium spp. that can affect domestic animal and human populations. In newborn ruminants, cryptosporidiosis is characterized by outbreaks of diarrhea, which can result in high morbidity and economic impact. The aim of t...

  13. Characteristics of a Novel Type of Bovine Cryptosporidium andersoni

    PubMed Central

    Satoh, Masaaki; Hikosaka, Kenji; Sasaki, Takako; Suyama, Yoshihisa; Yanai, Tokuma; Ohta, Minoru; Nakai, Yutaka

    2003-01-01

    We isolated oocysts that resemble Cryptosporidium andersoni from cattle grazing on a farm in Japan. The partial sequences of genes from the isolate were coincident with published sequences of genes of C. andersoni. Since the isolate was able to infect SCID mice, the isolate appears to be a novel type of C. andersoni. PMID:12514063

  14. TESTING METHODS FOR DETECTION OF CRYPTOSPORIDIUM SPP. IN WATER SAMPLES

    EPA Science Inventory

    A large waterborne outbreak of cryptosporidiosis in Milwaukee, Wisconsin, U.S.A. in 1993 prompted a search for ways to prevent large-scale waterborne outbreaks of protozoan parasitoses. Methods for detecting Cryptosporidium parvum play an integral role in strategies that lead to...

  15. PROBES FOR THE SPECIFIC DETECTION OF CRYPTOSPORIDIUM PARVUM

    EPA Science Inventory

    A probe set, consisting of two synthetic oligonucleotides each tagged with a fluorescent reporter molecule, has been developed for specific detection of Cryptosporidium parvum.Each probe strand detects ribosomal RNA from a range of isolates of this species, and the combination is...

  16. UTILITY OF SURROGATES FOR MEASURING CRYPTOSPORIDIUM OOCYST INFECTIVITY

    EPA Science Inventory

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

  17. UTILITY OF SURROGATES FOR MEASURING CRYPTOSPORIDIUM OOCYSTS INFECTIVITY

    EPA Science Inventory

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

  18. Subtyping Cryptosporidium ubiquitum, an emerging zoonotic pathogen in humans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryptosporidium ubiquitum is an emerging and important human pathogen with a broad range of mammalian hosts. Linkage between human and animal cases of C. ubiquitum has not been possible because of a lack of subtyping tools. In this study, we utilized the 60 kDa glycoprotein gene of C. ubiquitum to d...

  19. A sensitive method for detecting and genotyping Cryptosporidium parvum oocysts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryptosporidium parvum oocysts represent a considerable health risk to humans and animals because the parasite has a low infectious dose and usually exists in low numbers in environmental samples, which makes detection problematic. The purpose of this study was to evaluate Cryspovirus as a target f...

  20. Prevalence and species of Cryptosporidium in human populations worldwide

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A high prevalence of cryptosporidiosis has been reported in children under 5 years of age in sub-Saharan Africa, especially, but not exclusively, the malnourished and those positive for HIV infection. Infection rates are highest during the rainy season. Cryptosporidium hominis, which is transmitted ...

  1. Identification of Cryptosporidium from Dairy Cattle in Pahang, Malaysia.

    PubMed

    Hisamuddin, Nur Hazirah; Hashim, Najat; Soffian, Sharmeen Nellisa; Amin, Mohd Hishammfariz Mohd; Wahab, Ridhwan Abdul; Mohammad, Mardhiah; Isa, Muhammad Lokman Md; Yusof, Afzan Mat

    2016-04-01

    Cryptosporidium, a protozoan parasite, can cause cryptosporidiosis which is a gastrointestinal disease that can infect humans and livestock. Cattle are the most common livestock that can be infected with this protozoan. This study was carried out to determine the prevalence of Cryptosporidium infection in cattle in Kuantan, Pahang, Malaysia and to find out the association between the occurrence of infection and 3 different ages of cattle (calves less than 1 year, yearling, and adult cattle). The samples were processed by using formol-ether concentration technique and stained by modified Ziehl Neelsen. The results showed that 15.9% (24/151) of cattle were positive for Cryptosporidium oocysts. The occurrence of Cryptosporidium in calves less than 1 year was the highest with the percentage of 20.0% (11/55) followed by yearling and adult cattle, with the percentage occurrence of 15.6 % (7/45) and 11.8% (6/51), respectively. There was no significant association between the occurrence and age of cattle and presence of diarrhea. Good management practices and proper hygiene management must be taken in order to reduce the infection. It is highly important to control the infection since infected cattle may serve as potential reservoirs of the infection to other animals and humans, especially animal handlers. PMID:27180579

  2. Three sample preparation protocols for polymerase chain reaction based detection of Cryptosporidium parvum in environmental samples.

    PubMed

    Kostrzynska, M; Sankey, M; Haack, E; Power, C; Aldom, J E; Chagla, A H; Unger, S; Palmateer, G; Lee, H; Trevors, J T; De Grandis, S A

    1999-02-01

    Cryptosporidium parvum is a protozoan parasite responsible for an increasing number of outbreaks of gastrointestinal illness worldwide. In this report, we describe development of sample preparation protocols for polymerase chain reaction (PCR)-based detection of C. parvum in fecal material and environmental water samples. Two of these methods were found adequate for isolation of Cryptosporidium DNA from filtered water pellet suspensions. The first involved several filtration steps, immunomagnetic separation and freeze-thaw cycles. The second method involved filtration, addition of EnviroAmp lysis reagent, freeze-thaw cycles and precipitation of the DNA with isopropanol. Using nested PCR, we detected 100 oocysts/ml of filtered water pellet suspension, with either of the above sample preparation procedures. Nested PCR increased sensitivity of the assay by two to three orders of magnitude as compared to the primary PCR. The detection limit for seeded fecal samples was 10-fold higher than for filtered environmental water pellet suspension. Nested PCR results showed 62.4 and 91.1% correlation with immunofluorescence assay (IFA) for fecal samples and filtered environmental water pellet suspensions, respectively. This correlation decreased to 47.2% and 44.4%, respectively, when only IFA positive samples were analyzed. However, in fecal samples contaminated with a high number (> 10(5)/g) of C. parvum oocysts, this correlation was 100%. PMID:10076632

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

    PubMed Central

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

    1995-01-01

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

  4. Additional treatment of wastewater reduces endocrine disruption in wild fish--a comparative study of tertiary and advanced treatments.

    PubMed

    Baynes, Alice; Green, Christopher; Nicol, Elizabeth; Beresford, Nicola; Kanda, Rakesh; Henshaw, Alan; Churchley, John; Jobling, Susan

    2012-05-15

    Steroid estrogens are thought to be the major cause of feminization (intersex) in wild fish. Widely used wastewater treatment technologies are not effective at removing these contaminants to concentrations thought to be required to protect aquatic wildlife. A number of advanced treatment processes have been proposed to reduce the concentrations of estrogens entering the environment. Before investment is made in such processes, it is imperative that we compare their efficacy in terms of removal of steroid estrogens and their feminizing effects with other treatment options. This study assessed both steroid removal and intersex induction in adult and early life stage fish (roach, Rutilus rutilus). Roach were exposed directly to either secondary (activated sludge process (ASP)), tertiary (sand filtrated (SF)), or advanced (chlorine dioxide (ClO(2)), granular activated charcoal (GAC)) treated effluents for six months. Surprisingly, both the advanced GAC and tertiary SF treatments (but not the ClO(2) treatment) significantly removed the intersex induction associated with the ASP effluent; this was not predicted by the steroid estrogen measurements, which were higher in the tertiary SF than either the GAC or the ClO(2). Therefore our study highlights the importance of using both biological and chemical analysis when assessing new treatment technologies. PMID:22500691

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

    PubMed Central

    Kváč, Martin; Havrdová, Nikola; Hlásková, Lenka; Daňková, Tereza; Kanděra, Jiří; Ježková, Jana; Vítovec, Jiří; Sak, Bohumil; Ortega, Ynes; Xiao, Lihua; Modrý, David; Chelladurai, Jeba Rose Jennifer Jesudoss; Prantlová, Veronika; McEvoy, John

    2016-01-01

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

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

    PubMed

    Kváč, Martin; Havrdová, Nikola; Hlásková, Lenka; Daňková, Tereza; Kanděra, Jiří; Ježková, Jana; Vítovec, Jiří; Sak, Bohumil; Ortega, Ynes; Xiao, Lihua; Modrý, David; Chelladurai, Jeba Rose Jennifer Jesudoss; Prantlová, Veronika; McEvoy, John

    2016-01-01

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

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

    PubMed

    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 (P<.05), and was completely eliminated by day 28 pi. The mean weight gain of the P. granatum-treated mice was significantly higher than that of the infected/untreated controls throughout the study (P<.01). Histopathological analysis of ileal sections further supported the clinical and parasitological findings. The histological architecture 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

  8. Evaluation of a Combination Rapid Immunoassay for Detection of Giardia and Cryptosporidium Antigens

    PubMed Central

    Chan, Raymond; Chen, Jing; York, Mary K.; Setijono, Norman; Kaplan, Raymond L.; Graham, Fitzroy; Tanowitz, Herbert B.

    2000-01-01

    A combination cassette format nonenzymatic rapid immunoassay for detection of Giardia and Cryptosporidium antigens was evaluated by using 556 patient stool specimens from three clinical laboratories. This assay (Genzyme Diagnostics Contrast Giardia/Cryptosporidium), which can be used with fresh or formalin-fixed specimens, had unadjusted sensitivities and specificities of 96.1 and 98.5% for Giardia and 100 and 98.7% for Cryptosporidium, respectively, in this study. PMID:10618122

  9. Genetic characterization of Cryptosporidium isolates from ringed seals (Phoca hispida) in Northern Quebec, Canada.

    PubMed

    Santín, Mónica; Dixon, Brent R; Fayert, Ronald

    2005-06-01

    This study reports the molecular characterization of Cryptosporidium spp. isolates identified from intestinal contents of ringed seals (Phoca hispida) from Nunavik (Quebec, Canada). Cryptosporidium spp. fragments of 18S rRNA, HSP-70, and actin loci were amplified by PCR from seal intestinal contents. PCR-positive specimens were sequenced and compared with other Cryptosporidium species and genotypes reported previously. Sequence analysis showed the presence of C. muris and 2 novel genotypes in ringed seals. PMID:16108576

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

    PubMed

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

    2016-08-15

    Cryptosporidium is a protozoan parasite that infects a wide range of hosts, yet relatively little is known about the epidemiology of cryptosporidiosis in fish. Here we report a disseminated Cryptosporidium infection in a male Koi carp (Cyprinus carpio), with parasite stages identified deep within the epithelium of the intestine, kidneys, spleen, liver and gills causing severe granulomatous inflammatory lesions. Molecular characterization at two loci; 18S ribosomal RNA (rRNA) and actin, revealed this to be a novel Cryptosporidium genotype, most closely related to Cryptosporidium molnari. PMID:27514884

  11. First Molecular Characterization of Cryptosporidium spp. Infecting Buffalo Calves in Brazil.

    PubMed

    Aquino, Monally C C; Widmer, Giovanni; Zucatto, Anaiza S; Viol, Milena A; Inácio, Sandra V; Nakamura, Alex A; Coelho, Willian M D; Perri, Silvia H V; Meireles, Marcelo V; Bresciani, Katia D S

    2015-01-01

    With the aim of determining the occurrence of Cryptosporidium spp., 222 fecal samples were collected from Murrah buffalo calves aged up to 6 mo. Fecal DNA was genotyped with a nested polymerase chain reaction targeting the 18S rRNA gene and sequencing of the amplified fragment. Nested 18S PCR was positive for 48.2% of the samples. Sequence analysis showed that the most frequent species in these animals was Cryptosporidium ryanae, which was present in buffalo calves as young as 5 d. The zoonotic species Cryptosporidium parvum was detected in one animal. An uncommon Cryptosporidium 18S genotype was found in buffaloes. PMID:25941018

  12. Molecular characterization of Cryptosporidium spp. from fecal samples of birds kept in captivity in Brazil.

    PubMed

    Nakamura, Alex Akira; Simões, Daniel Castendo; Antunes, Rômulo Godik; da Silva, Deuvânia Carvalho; Meireles, Marcelo Vasconcelos

    2009-12-01

    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). PMID:19683397

  13. Decrease in the acrylamide content in canned coffee by heat treatment with the addition of cysteine.

    PubMed

    Narita, Yusaku; Inouye, Kuniyo

    2014-12-17

    Acrylamide (AA) is classified as a Group 2A carcinogen according to the International Agency for Research on Cancer. Although coffee contains a small amount of AA, it is a popular beverage worldwide. Approximately 10 billion canned coffees are consumed each year in Japan. In this study, we investigated how to decrease AA contained in canned coffee by modifying the heat treatment used for sterilization during the manufacturing process. The AA content of both types of canned coffee (black and milk) was decreased by approximately 95% by heat treatment with adding cysteine at 121 °C for 6 min. The content was also decreased by heat treatment with dithiothreitol, although that with cystine had no effect. Therefore, it is shown that thiol groups in cysteine and dithiothreitol might play an important role in decreasing the AA content. PMID:25420187

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

    EPA Science Inventory

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

  15. Additive, Multi-Component Treatment of Emerging Refusal Topographies in a Pediatric Feeding Disorder

    ERIC Educational Resources Information Center

    Sharp, William G.; Jaquess, David L.; Bogard, Jennifer D.; Morton, Jane F.

    2010-01-01

    This case study describes inter-disciplinary treatment of chronic food refusal and tube dependency in a 2-year-old female with a pediatric feeding disorder. Evidence-based behavioral components--including escape extinction (EE), differential reinforcement of alterative mealtime behavior (DRA), and stimulus fading--were introduced sequentially as…

  16. UASB Treatment of Methanolic Pulp Wastewater with Addition of Waste Starch and Incinerated Ash

    NASA Astrophysics Data System (ADS)

    Takahashi, Shintaro; Kobaysashi, Takuro; Li, Yu-You; Harada, Hideki

    The pulp wastewater consists mainly of methanol. It is expected to treat using upflow anaerobic sludge blanket (UASB) process. Paper manufactories also produce waste starch and incinerated ash. The integrated treating for these wastes is desirable. In this study, two UASB reactors were operated to treat pulp wastewater with addition of waste starch and with addition of incinerated ash, receptively. Continuous operations of a UASB reactor treating pulp wastewater with addition of waste starch (PS reactor) and a UASB reactor treating pulp wastewater with addition of incinerated ash (PA reactor) , were investigated at mesophilic conditions. The PS reactor performed well with an average 93.7% total CODCr and 97.3% soluble CODCr removal efficiency in average at a maximum volumetric loading rate (VLR) of 16.0 kgCOD/m3/d. The PA reactor was also successfully operated with an average 95.3% total CODCr and 97.5% soluble CODCr removal efficiency in average at a maximum VLR of 14.6 kgCOD/m3/d. Successfully developed granules were obtained after over 140 days of operation in both reactors, and the granules were 1 to 2 mm in mean diameter. Microbial analysis revealed the genus Methanomethylovorans was predominant in the granules of both reactors.

  17. Hydrothermal treatment of incineration fly ash for PCDD/Fs decomposition: the effect of iron addition.

    PubMed

    Chen, De-Zhen; Hu, Yu-Yan; Zhang, Peng-Fei

    2012-12-01

    The catalytic effect of Fe addition on the decomposition of polychlorinated dibenzo-dioxins and polychlorinated dibenzofurans (PCDD/Fs) contained in municipal solid waste incineration (MSWI) fly ash during the hydrothermal process was investigated. Influencing factors, such as Fe addition mode, reaction time and cooling procedure after reaction, were tested to evaluate their effects. Experimental results indicated that Fe addition in the form of a mixture of ferrous sulphate and ferric sulphate enhanced decomposition of PCDD/Fs contained in the MSWI fly ash, particularly for the decomposition of 2,3,7,8-tetrachlorodibenzo-dioxin and 2,3,7,8-tetrachlorodibenzo-furan under the reaction temperature of 563 K. The decomposition rate of PCDD/Fs reached 90.33% by international toxicity equivalent (I-TEQ) when Fe was added as a mixture of ferrous and ferric sulphates by 5% (wt/wt) with the Fe (III)/Fe (II) ratio being 2; without Fe addition, the decomposition rate of PCDD/Fs was only 46.17% by I-TEQ in the same process. Fe addition in the form of ferrous sulphate alone also showed an enhancing effect on PCDD/Fs decomposition, but the associated decomposition rates were relatively lower, suggesting iron oxides formed from the mixture of ferric and ferrous sulphates are more favourable catalysts. At the same time, the cooling procedure after the hydrothermal reaction became more flexible if Fe was added in the form of a mixture of ferric and ferrous sulphates. Although a longer reaction time was helpful to increase decomposition rates of PCDD/Fs, 1 h was proved to be a reasonable time under this condition. PMID:23437649

  18. Is There an Additional Value of Inpatient Treatment for Patients with Atopic Dermatitis?

    PubMed

    van der Schaft, Jorien; Keijzer, Welmoed W; Sanders, Koos J G; de Groot, Jette J C; van den Bersselaar, Donna L C M; van Os-Medendorp, Harmieke; van Doorn-Op den Akker, Margreet M; Bruijnzeel-Koomen, Carla A F M; de Bruin-Weller, Marjolein S

    2016-08-23

    An inpatient treatment and education programme has been developed for patients with difficult to control atopic dermatitis (AD), with the aim of achieving adequate self-management and long-term disease control. This observational study included adult patients diagnosed with difficult to control AD, admitted for a structured inpatient treatment and education programme. The primary outcome was the Six Area, Six Sign Atopic Dermatitis (SASSAD) score. In total, 79 patients (mean ± SD age 38.8 ± 17.1 years) were included. The median duration of hospitalization was 11 days (interquartile range 8-14). The mean percentage decrease in SASSAD score between admission and discharge was 60.7%, of which 64 (81.0%) patients achieved SASSAD50. The mean percentage decrease in SASSAD score was 69.0% during follow-up, of which 63 (79.7%) patients still had a SASSAD50. In the majority of these patients with difficult to control AD the admission resulted in sustained disease control. This could be achieved by optimization of treatment with topical corticosteroids. PMID:26983375

  19. Ultraviolet and pulsed electric field treatments have additive effect on inactivation of E. coli in apple juice.

    PubMed

    Gachovska, T K; Kumar, S; Thippareddi, H; Subbiah, J; Williams, F

    2008-11-01

    Apple juice inoculated with Escherichia coli ATCC 23472 was processed continuously using either ultraviolet (UV), high-voltage pulsed electric field (PEF), or a combination of the PEF and UV treatment systems. Apple juice was pumped through either of the systems at 3 flow rates (8, 14, and 20 mL/min). E. coli was reduced by 3.46 log CFU/mL when exposed in a 50 cm length of UV treatment chamber at 8 mL/min (2.94 s treatment time with a product temperature increase of 13 degrees C). E. coli inactivation of 4.87 log CFU/mL was achieved with a peak electric field strength of 60 kV/cm and 11.3 pulses (average pulse width of 3.5 mus, product temperature increased to 52 degrees C). E. coli reductions resulting from a combination treatment of UV and PEF applied sequentially were evaluated. A maximum E. coli reduction of 5.35 log CFU/mL was achieved using PEF (electrical field strength of 60 kV/cm, specific energy of 162 J/mL, and 11.3 pulses) and UV treatments (length of 50 cm, treatment time of 2.94 s, and flow rate of 8 mL/min). An additive effect was observed for the combination treatments (PEF and UV), regardless of the order of treatment (P > 0.05). E. coli reductions of 5.35 and 5.30 log CFU/mL with PEF treatment (electrical field strength of 60 kV/cm, specific energy of 162 J/mL, and 11.3 pulses) followed by UV (length of 30 cm, treatment time of 1.8 s, and flow rate of 8 mL/min) and UV treatment followed by PEF (same treatment conditions), respectively. No synergistic effect was observed. PMID:19021811

  20. Detection and characterization of a Cryptosporidium isolate from a southern elephant seal (Mirounga leonina) from the Antarctic peninsula.

    PubMed

    Rengifo-Herrera, C; Ortega-Mora, L M; Gómez-Bautista, M; García-Moreno, F T; García-Párraga, D; Castro-Urda, J; Pedraza-Díaz, S

    2011-02-01

    The presence of Cryptosporidium and Giardia in 221 fecal samples from different species of Antarctic pinnipeds was investigated by immunofluorescence microscopy and PCR. Cryptosporidium, a skunk-like genotype, was detected only in a southern elephant seal. Giardia was not detected. This is the first report of a Cryptosporidium sp. in Antarctic marine mammals. PMID:21169427

  1. Diarrhea in lambs experimentally infected with Cryptosporidium isolated from calves.

    PubMed

    Tzipori, S; Angus, K W; Gray, E W; Campbell, I; Allan, F

    1981-08-01

    Two to 5 days after oral inoculation with Cryptosporidium obtained from infected calves, 7 newborn specific-pathogen-free lambs became depressed and anorectic and developed diarrhea. Four moribund lambs were killed within 3 days after onset of illness, and the 3 other lambs died after protracted intermittent diarrhea and reduced milk intake, which lasted up to 14 days. The small and large intestines were heavily infected with Cryptosporidium, with the terminal portion of the ileum being the most severely affected. Histologic examination revealed villous atrophy, with considerable fusion and epithelial cross-bridging between villi. Lambs infected at 5 to 20 days of age had less severe clinical signs of disease, with intermittent diarrhea, reduced milk intake, and growth retardation. Lambs infected at 30 days of age became infected with the organism, but did not develop clinical signs of disease or growth retardation. PMID:7294476

  2. Possible zoonotic transmission of Cryptosporidium felis in a household

    PubMed Central

    Beser, Jessica; Toresson, Linda; Eitrem, Rickard; Troell, Karin; Winiecka-Krusnell, Jadwiga; Lebbad, Marianne

    2015-01-01

    In humans, the risk of contracting cryptosporidiosis caused by Cryptosporidium felis is considered to be relatively low, and most of the confirmed cases have been observed in immunocompromised patients. Both anthroponotic and zoonotic transmission routes have been suggested. Here, we report a case of suspected zoonotic transmission of C. felis from a cat to a human. The cat developed diarrhea several months before such symptoms were displayed by its owner, a 37-year-old immunocompetent woman. The presence of identical C. felis SSU rRNA, HSP70, and COWP gene sequences was verified in both hosts. In conclusion, it is highly probable that the cat was the initial source of infection and not the opposite. Our results show that Cryptosporidium infection can be transmitted from pets to humans and that molecular analysis is needed to confirm the identity of the oocysts. PMID:26446304

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

    PubMed

    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. PMID:19500463

  4. [Cryptosporidium parvum Gastroenteritis in a Patient with Renal Transplantation].

    PubMed

    Çetinkaya, Ülfet; Dursun, İsmail; Kuk, Salih; Şahin, İzzet; Yazar, Süleyman

    2015-09-01

    In this study, a case who starting abundant watery diarrhea on the 14th day of renal transplantation is presented. Stool sample was analyzed for Cryptosporidium spp. by carbol fuchsin staining method, copro-ELISA and nested polimeraze chain reaction (PCR). From sample found positive by Carbol-fuchsin staining method and Copro-ELISA, DNA sequence analysis was performed, gel-purified from amplicon obtained by nested PCR. As a result of DNA sequence analysis was determined to be Cryptosporidium parvum. Although C. parvum is a rare causative agent of gastroenteritis it can be cause serious clinical diarrhea solid organ transplantation patient. As a result, also C.parvum must be considered as a causative agent of diarrhea occurring after organ transplantation. PMID:26470932

  5. Cryptosporidium avium n. sp. (Apicomplexa: Cryptosporidiidae) in birds.

    PubMed

    Holubová, Nikola; Sak, Bohumil; Horčičková, Michaela; Hlásková, Lenka; Květoňová, Dana; Menchaca, Sarah; McEvoy, John; Kváč, Martin

    2016-06-01

    The morphological, biological, and molecular characteristics of Cryptosporidium avian genotype V are described, and the species name Cryptosporidium avium is proposed to reflect its specificity for birds under natural and experimental conditions. Oocysts of C. avium measured 5.30-6.90 μm (mean = 6.26 μm) × 4.30-5.50 μm (mean = 4.86 μm) with a length to width ratio of 1.29 (1.14-1.47). Oocysts of C. avium obtained from four naturally infected red-crowned parakeets (Cyanoramphus novaezealandiae) were infectious for 6-month-old budgerigars (Melopsittacus undulatus) and hens (Gallus gallus f. domestica). The prepatent periods in both susceptible bird species was 11 days postinfection (DPI). The infection intensity of C. avium in budgerigars and hens was low, with a maximum intensity of 5000 oocysts per gram of feces. Oocysts of C. avium were microscopically detected at only 12-16 DPI in hens and 12 DPI in budgerigars, while PCR analyses revealed the presence of specific DNA in fecal samples from 11 to 30 DPI (the conclusion of the experiment). Cryptosporidium avium was not infectious for 8-week-old SCID and BALB/c mice (Mus musculus). Naturally or experimentally infected birds showed no clinical signs of cryptosporidiosis, and no pathology was detected. Developmental stages of C. avium were detected in the ileum and cecum using scanning electron microscopy. Phylogenetic analyses based on small subunit rRNA, actin, and heat shock protein 70 gene sequences revealed that C. avium is genetically distinct from previously described Cryptosporidium species. PMID:26905074

  6. Addition of a Magnetite Layer onto a Polysulfone Water Treatment Membrane to Enhance Virus Removal

    NASA Astrophysics Data System (ADS)

    Raciny, Isabel

    The applicability of low-pressure membranes systems in distributed (point of use) water treatment is hindered by, among other things, their inability to remove potentially harmful viruses and ions via size exclusion. According to the USEPA and the Safe Drinking Water Act, drinking water treatment processes must be designed for 4-log virus removal. Batch experiments using magnetite nanoparticle (nano-Fe3O4) suspensions and water filtration experiments with Polysulfone (PSf) membranes coated with nano-Fe3O 4 were conducted to assess the removal of a model virus (bacteriophage MS2). The membranes were coated via a simple filtration protocol. Unmodified membranes were a poor adsorbent for MS2 bacteriophage with less than 0.5-log removal, whereas membranes coated with magnetite nanoparticles exhibited a removal efficiency exceeding 99.99% (4-log). Thus, a cartridge of PSf membranes coated with nano-Fe3O4 particles could be used to remove viruses from water. Such membranes showed negligible iron leaching into the filtrate, thus obviating concern about colored water. Further research is needed to reduce the loss of water flux caused by coating.

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

    PubMed

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

    2014-01-01

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

  8. Diarrhea due to Cryptosporidium infection in artificially reared lambs.

    PubMed Central

    Tzipori, S; Angus, K W; Campbell, I; Clerihew, L W

    1981-01-01

    Severe diarrhea which lasted 7 to 12 days occurred in 40 of 48 artificially reared lambs within 5 to 12 days of birth, and 16 of them died. Of 16 diarrheic fecal samples examined, 10 contained Cryptosporidium oocysts and 1 contained rotavirus, but no other known enteropathogen was detected. Upon histological examination, cryptosporidia were found in the ilea of three affected lambs, and in one of them, villous atrophy and fusion, with epithelial cross-bridging between villi, were present in distal small intestine. Diarrhea was induced in two specific pathogen-free lambs by oral inoculation with fecal homogenate containing Cryptosporidium oocysts. Both the small and large intestines became infected with the organism, and associated lesions included stunting, fusion, and deformities of villi in the distal small intestine, with replacement of columnar enterocytes by immature cuboidal cells. Subclinical infections were induced in newborn specific pathogen-free mice and rats. Judged by these data, the lamb-derived Cryptosporidium sp. is similar to those recovered from calves, deer, and humans. Images PMID:7263849

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

    PubMed

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

    1996-10-01

    A recent report suggested that an isolate of Cryptosporidium parvum had established infections in fish, amphibians, and reptiles and raises concern that animals other than mammals might be a potential source of waterborne Cryptosporidium oocysts. To test this possibility, viable C. parvum oocysts, infectious for neonatal BALB/c mice, were delivered by gastric intubation to bluegill sunfish, poison-dart frogs, African clawed frogs, bearded dragon lizards, and corn snakes. Histological sections of the stomach, jejunum, ileum, and cloaca prepared from tissues collected on days 7 and 14 postinoculation (PI) were negative for Cryptosporidium developmental stages. However, inoculum-derived oocysts were detectable by fluorescein-labeled monoclonal antibody in feces of inoculated animals from day 1 to day 12 PI in fish and frogs, and up to day 14 PI in lizards. Snakes did not defecate for 14 days PI. Impression smears taken at necropsy on days 7 and 14 PI revealed C. parvum oocysts in the lumen of the cloaca of 2 fish and 1 lizard on day 7 PI only. Because tissue stages of the pathogen were not found, it appears that C. parvum was not heterologously transmitted to lower vertebrates. Under certain circumstances, however, such as after the ingestion of C. parvum-infected prey, lower vertebrates may disseminate C. parvum oocysts in the environment. PMID:8885883

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

    PubMed Central

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

    2014-01-01

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

  11. Effectiveness of additional supervised exercises compared with conventional treatment alone in patients with acute lateral ankle sprains: systematic review

    PubMed Central

    van Ochten, John; Luijsterburg, Pim A J; van Middelkoop, Marienke; Koes, Bart W; Bierma-Zeinstra, Sita M A

    2010-01-01

    Objective To summarise the effectiveness of adding supervised exercises to conventional treatment compared with conventional treatment alone in patients with acute lateral ankle sprains. Design Systematic review. Data sources Medline, Embase, Cochrane Central Register of Controlled Trials, Cinahl, and reference screening. Study selection Included studies were randomised controlled trials, quasi-randomised controlled trials, or clinical trials. Patients were adolescents or adults with an acute lateral ankle sprain. The treatment options were conventional treatment alone or conventional treatment combined with supervised exercises. Two reviewers independently assessed the risk of bias, and one reviewer extracted data. Because of clinical heterogeneity we analysed the data using a best evidence synthesis. Follow-up was classified as short term (up to two weeks), intermediate (two weeks to three months), and long term (more than three months). Results 11 studies were included. There was limited to moderate evidence to suggest that the addition of supervised exercises to conventional treatment leads to faster and better recovery and a faster return to sport at short term follow-up than conventional treatment alone. In specific populations (athletes, soldiers, and patients with severe injuries) this evidence was restricted to a faster return to work and sport only. There was no strong evidence of effectiveness for any of the outcome measures. Most of the included studies had a high risk of bias, with few having adequate statistical power to detect clinically relevant differences. Conclusion Additional supervised exercises compared with conventional treatment alone have some benefit for recovery and return to sport in patients with ankle sprain, though the evidence is limited or moderate and many studies are subject to bias. PMID:20978065

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

    PubMed

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

    2010-06-01

    A field study in Valencia, Spain, was done to determine the occurrence of Giardia and Cryptosporidium on salad products that are frequently eaten raw, such as lettuces and Chinese cabbage, and in irrigation waters. Four water samples were taken weekly 1 month before harvesting the vegetables. All water samples were analyzed using techniques included in the U.S. Environmental Protection Agency Method 1623. Standard methods for detecting protozoan parasites on salad vegetables are not available. Published techniques for the isolation of parasites from vegetables generally have low and variable recovery efficiencies. In this study, vegetables were analyzed using a recently reported method for detection of Cryptosporidium oocysts and Giardia cysts on salad products. The waters tested were positive for both Cryptosporidium and Giardia. Of 19 salad products studied, we observed Cryptosporidium in 12 samples and Giardia in 10 samples. Recoveries of the Texas Red-stained Cryptosporidium and Giardia, which were used as internal controls, were 24.5% +/- 3.5% for Cryptosporidium and 16.7% +/- 8.1% for Giardia (n = 8). This study provides data on the occurrence of Cryptosporidium and Giardia in salad products in Spain. The method was useful in the detection of Cryptosporidium oocysts and Giardia cysts on the vegetables tested, and it provides a useful analytical tool for occurrence monitoring. PMID:20537274

  13. Cryptosporidium ryanae n.sp. (Apicomplexa:Cryptosporidiidae)in cattle (Bos Taurus)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A new species, Cryptosporidium ryanae, is described from cattle. Oocysts of C. ryanae, previously identified as the Cryptosporidium deer-like genotype and recorded as such in GenBank (AY587166, EU203216, DQ182597, AY741309, and DQ871345), are morphologically similar to those of C. parvum and C. bovi...

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

    EPA Science Inventory

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

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

    EPA Science Inventory

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

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

    EPA Science Inventory

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

  17. The first large scale molecular study of Cryptosporidium and Giardia in horses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The prevalence of species and genotypes of Giardia and Cryptosporidium in horses is poorly known. The present study examined feces from 195 horses, 1 month to 17 years of age, in 4 locations in Colombia. Prevalence and age distribution of Giardia and Cryptosporidium were determined by PCR. All PCR p...

  18. Wide distribution of Cryptosporidium bovis and the deer-like genotype in bovines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We recently reported that on 14 dairy farms from Vermont to Florida ~85% of pre-weaned dairy calves were infected with zoonotic Cryptosporidium parvum whereas only 1-2% of post-weaned calves and 1-2 year-old heifers were infected with this species. Cryptosporidium bovis and the deer-like genotype w...

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

    PubMed

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

    2015-06-01

    To investigate the prevalence and assess the zoonotic transmission burden of Cryptosporidium species in domestic pigeons in Guangdong Province, Southern China, 244 fecal samples were collected from four pigeon breeding farms between June 2012 and March 2013. Cryptosporidium oocysts were purified by Sheather's sugar flotation technique and characterized by DNA sequencing of small subunit ribosomal RNA (SSU rRNA) gene. Cryptosporidium species were determined by comparison of sequences with corresponding Cryptosporidium sequences in GenBank and phylogenetic analysis using neighbor-joining (NJ) in MEGA5.2. The overall prevalence of Cryptosporidium infection in domestic pigeons in Guangdong Province was 0.82% (2/244). Two Cryptosporidium species, namely Cryptosporidium baileyi and Cryptosporidium meleagridis, were identified in Huizhou and Chaozhou farm, respectively. These findings confirmed the existence of C. meleagridis infection in domestic pigeons in China for the first time and provided base-line information for further studies to evaluate the public health risk from pigeon to human. PMID:25773186

  20. Molecular investigation of Cryptosporidium in small caged pets in northeast China: host specificity and zoonotic implications.

    PubMed

    Li, Qiao; Li, Lu; Tao, Wei; Jiang, Yanxue; Wan, Qiang; Lin, Yongchao; Li, Wei

    2016-07-01

    This study screened 151 pet-derived fecal specimens randomly collected from four commercial markets in northeast China for the presence of Cryptosporidium by genus-specific nested PCRs of the small subunit rRNA gene. Of these, 14 specimens (9.3 %) from nine species of birds, two types of rodents, and a hedgehog were positive for Cryptosporidium. Sequence analysis on the PCR-positive isolates facilitated identification of three Cryptosporidium species (C. baileyi, C. galli, and C. ubiquitum) and two Cryptosporidium genotypes (ferret genotype and avian genotype V). The study birds were affected predominantly with bird-specific C. baileyi (Atlantic canary, budgerigar, crested myna, rock dove, and silky fowl), C. galli (Chinese hwamei), and Cryptosporidium avian genotype V (Fischer's lovebird and rosy-faced lovebird). Cryptosporidium ferret genotype previously considered rodent-adapted was identified in three specimens from budgerigar, chipmunk, and red squirrel. Two specimens collected from common hill myna and hedgehog were positive for C. ubiquitum. The species of birds that can be colonized by Cryptosporidium were extended. Moreover, the data expanded the host range of Cryptosporidium ferret genotype and C. ubiquitum, especially the birds. The carriage of zoonotic C. ubiquitum in small caged pets is of public health importance. PMID:27107987

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

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

    EPA Science Inventory

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

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

    EPA Science Inventory

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

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

    EPA Science Inventory

    Since the first identified Cryptosporidium outbreaks occurred in the 1980s and the massive 1993 Milwaukee, WI outbreak affected more than 400,000 people, the concern over the public health risks related to protozoan pathogens Cryptosporidium and Giardia has g...

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

    PubMed

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

    2015-10-01

    Cryptosporidium and Enterocytozoon bieneusi are the most prevalent protist pathogens responsible for inducing human and animal diseases worldwide. The aim of the present work was to determine the occurrence of Cryptosporidium spp. and E. bieneusi in pet chinchillas in China. One hundred forty fecal samples were collected from four cities: Beijing, Zhengzhou, Anyang and Guiyang. They were then examined with PCR amplification of the small subunit ribosomal RNA (SSU rRNA) of Cryptosporidium spp. and the internal transcribed spacer (ITS) of the ribosomal RNA of E. bieneusi. The infection rates for Cryptosporidium spp. and E. bieneusi were 10.0% and 3.6%, respectively. Sequence analysis of SSU rRNA gene products identified two Cryptosporidium spp., Cryptosporidium ubiquitum (n=13) and Cryptosporidium parvum (n=1). Subtyping with the 60-kDa glycoprotein (gp60) gene showed that all C. ubiquitum isolates belonged to zoonotic subtype family XIId, while the subtype of the C. parvum isolate could not be identified. Two E. bieneusi genotypes were identified in five samples, zoonotic genotypes BEB6 (n=3) and D (n=2). This is the first report of C. ubiquitum and C. parvum, and E. bieneusi in chinchillas. This result indicates that pet chinchillas may be a potential source of human infection with Cryptosporidium spp. and E. bieneusi. PMID:25988830

  6. Prevalence of Cryptosporidium infection in captive lesser panda (Ailurus fulgens) in China.

    PubMed

    Wang, Tao; Chen, Zuqin; Yu, Hua; Xie, Yue; Gu, Xiaobing; Lai, Weiming; Peng, Xuerong; Yang, Guangyou

    2015-02-01

    Cryptosporidium is a global epidemic parasite and one of the most important intestinal pathogens causing diarrhea in animals and humans. Despite extensive research on this parasite group, little is known about rates of Cryptosporidium infection in lesser pandas. In this study, we use molecular diagnostic tools to detect Cryptosporidium infections and identify Cryptosporidium species in the lesser panda. Using a PCR approach, we sequenced the 18S rRNA gene in fecal samples collected from 110 captive lesser pandas held throughout China (approximately one third of the captive population). We determined Cryptosporidium species via a BLAST comparison of our sequences against those of published Cryptosporidium sequences available in GenBank and subsequent phylogenetic analysis. We report that captive lesser pandas were infected with a single Cryptosporidium species, Cryptosporidium andersoni, at a prevalence of 6.36 % (7/110). The present investigation revealed the existence of C. andersoni infection in captive lesser panda and suggested that proper control measures should be taken carefully to protect the welfare of zoo workers and visitors. PMID:25563613

  7. Cryptosporidium species detected in calves and cattle in Dagoretti, Nairobi, Kenya.

    PubMed

    Kang'ethe, Erastus K; Mulinge, Erastus K; Skilton, Robert A; Njahira, Moses; Monda, Joseph G; Nyongesa, Concepta; Mbae, Cecilia K; Kamwati, Stanley K

    2012-09-01

    A total of 1,734 cattle faecal samples from 296 dairy-keeping households were collected from urban settings in Nairobi, Kenya. Modified Ziehl-Neelsen staining method and an immunofluorescence assay were used to identify those samples with Cryptosporidium oocyst infection. Oocysts from positive faecal samples were isolated by Sheather's sucrose flotation method and picked from the concentrate using cover slips. Genomic DNA was extracted from 124 of the faecal samples that were positive for Cryptosporidium and was used as template for nested PCR of the 18S rRNA gene. Twenty-five samples (20 %) were PCR-positive for Cryptosporidium, and 24 of the PCR products were successfully cloned and sequenced. Sequence and phylogenetic analysis identified 17 samples (68 %) as Cryptosporidium parvum-like, four samples (16 %) as Cryptosporidium ryanae, three samples (12 %) as Cryptosporidium andersoni and one sample (4 %) as Cryptosporidium hominis. To the best of our knowledge, this is the first genotyping study to report C. parvum-like, C. andersoni and C. hominis in cattle from Kenya. The results of this study show Cryptosporidium infections in calves and cattle may be potential zoonotic reservoirs of the parasite that infects humans. PMID:22797974

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

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

    EPA Science Inventory

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

  10. Gastroenteritis caused by the Cryptosporidium hedgehog genotype in an immunocompetent man.

    PubMed

    Kváč, Martin; Saková, Kamila; Kvĕtoňová, Dana; Kicia, Marta; Wesołowska, Maria; McEvoy, John; Sak, Bohumil

    2014-01-01

    The Cryptosporidium hedgehog genotype, which has been reported previously in hedgehogs and horses, was identified as the cause of the diarrheal disease cryptosporidiosis in an immunocompetent man in the Czech Republic. This is the first report of human illness caused by the Cryptosporidium hedgehog genotype. PMID:24131692

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

    EPA Science Inventory

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

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

    PubMed

    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) concentrations<250 mg/L and hydraulic retention times (HRTs) between 23 and 180 days were observed. Laboratory (oo)cysts inactivation studies were performed in bench-scale digesters, which were maintained under conditions similar to those observed in the field. Apparent first-order inactivation rate coefficients for Giardia 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

  13. Outcome following addition of peroneus brevis tendon transfer to treatment of acquired posterior tibial tendon insufficiency.

    PubMed

    Song, S J; Deland, J T

    2001-04-01

    The flexor digitorum longus, the tendon most often used for transfer in posterior tibial tendon insufficiency, is one-half to one-third the size of the posterior tibial tendon. Occasionally it may be particularly small or may have been previously used for transfer. In these cases, the senior author has felt that the addition of a transfer of the Peroneus Brevis (PBr) tendon may be helpful in maintaining sufficient tendon and muscle mass to rebalance the foot. Thirteen patients who underwent this procedure were retrospectively identified and matched by age and length of follow-up to patients who underwent a more standard tendon transfer operation minus the addition of the PBr transfer. Pain and functional status were then assessed by the American Orthopaedic Foot and Ankle Society's ankle/hindfoot rating scale. Each patient was tested by an independent physical therapist to evaluate inversion and eversion strength. The mean duration of follow-up was 20.6 months (12 to 34 months). The average AOFAS score of the PBr group was 75.8 compared to 71.5 for the standard control group. There was no significant difference between the groups when inversion or eversion strengths were compared. Inversion strength and eversion strength was rated good or excellent (4 or 5) in 12 out of 13 of the PBr transfer group patients. No major complications were encountered in either group. Although it does not increase inversion strength, a PBr transfer can be used to augment a small FDL without causing significant eversion weakness. This can be useful when the FDL is particularly small or in revision surgery. PMID:11354442

  14. Role of the addition of cabergoline to the management of acromegalic patients resistant to longterm treatment with octreotide LAR.

    PubMed

    Vilar, Lucio; Azevedo, Monalisa F; Naves, Luciana Ansaneli; Casulari, Luiz Augusto; Albuquerque, José Luciano; Montenegro, Renan M; Montenegro, Renan M; Figueiredo, Patricia; Nascimento, Gilvan C; Faria, Manuel S

    2011-06-01

    The aim of this prospective open trial was to evaluate the efficacy in normalizing IGF-I levels of the addition of cabergoline to the treatment of acromegalic patients partially responsive to Octreotide-LAR (OCT-LAR), a long acting somatotastin analog (SSA). Fifty-two patients who did not achieve hormonal control after longterm therapy (at least, 12 months) with OCT-LAR (30 mg every 28 days intramuscularly) were given cabergoline in addition to the SSA treatment. Normalization of IGF-I levels was achieved in 40.4% of patients by 6 months after the addition of cabergoline (1.0-3.0 mg/week; mean, 2.19 ± 0.64), and these patients were considered responsive. Compared to non-responsive subjects, responsive patients had significantly lower mean %ULNR-IGF-I and GH levels. However, the rate of hyperprolactinemia and positive immunohistochemical staining for PRL was similar in both groups, before the addition of cabergoline. Responsive patients were followed for at least 12 months on combination treatment and persisted with normal IGF-I levels. Patients with baseline %ULNR IGF-I up to 220% and/or GH up to 5 ng/ml were those who benefited the most from combination treatment. No patients with %ULNR-IGF-I>250% reached normalization of IGF-I levels. Our findings demonstrated that the addition of cabergoline, even at relatively low doses, is effective in both short- and long-term control of IGF-I levels in acromegalic patients partially responsive to octreotide LAR, particularly in those with mild/moderately elevated GH/IGF-levels, irrespective of prolactin status. PMID:21104199

  15. The additive and interactive effects of parenting style and temperament in obese youth seeking treatment

    PubMed Central

    Zeller, MH; Boles, RE; Reiter-Purtill, J

    2009-01-01

    Objective To examine maternal parenting behaviors, child temperament and their potential interactions in families of obese children and demographically similar families of nonoverweight children. Design A total of 77 obese youth (M body mass index (BMI) z-score values, zBMI = 2.4; ages 8–16, 59% female, 50% African American) and their parents were recruited from a pediatric weight management clinic and compared to 69 families of nonoverweight youth (M zBMI = − 0.03). Comparison youth were classmates of each obese participant matched on gender, race and age. Measurements Maternal report of child temperament, parenting style and anthropometric assessments were obtained. Results Compared to nonoverweight youth, mothers of obese youth described their child as having a more difficult temperament and their parenting style as lower in behavioral control. A logistic regression model indicated that difficult temperament, lower behavioral control and the interaction of low maternal warmth and difficult child temperament were associated with increased odds of a child being classified as obese. Conclusions Treatment-seeking obese youth and their parents are characterized by different parent and child factors when compared to nonoverweight comparison families. These findings direct investigators to test more complex models of the relation between parent and child characteristics and their mutual role in the weight-related behavior change process. PMID:18698318

  16. Is Correction of Iron Deficiency a New Addition to the Treatment of the Heart Failure?

    PubMed Central

    Silverberg, Donald S.; Wexler, Dov; Schwartz, Doron

    2015-01-01

    Anemia is present in about 40% of heart failure (HF) patients. Iron deficiency (ID) is present in about 60% of the patients with anemia (about 24% of all HF patients) and in about 40% of patients without anemia (about 24% of all HF patients). Thus ID is present in about half the patients with HF. The ID in HF is associated with reduced iron stores in the bone marrow and the heart. ID is an independent risk factor for severity and worsening of the HF. Correction of ID with intravenous (IV) iron usually corrects both the anemia and the ID. Currently used IV iron preparations are very safe and effective in treating the ID in HF whereas little information is available on the effectiveness of oral iron. In HF IV iron correction of ID is associated with improvement in functional status, exercise capacity, quality of life and, in some studies, improvement in rate of hospitalization for HF, cardiac structure and function, and renal function. Large long-term adequately-controlled intervention studies are needed to clarify the effect of IV iron in HF. Several heart associations suggest that ID should be routinely sought for in all HF patients and corrected if present. In this paper we present our approach to diagnosis and treatment of iron deficiency in heart failure. PMID:26096008

  17. Effect of Ag nanoparticle addition and ultrasonic treatment on a stable TiO2 nanofluid.

    PubMed

    Chakraborty, Subhrakanti; Mukherjee, Jonaki; Manna, Manindra; Ghosh, Pampa; Das, Sumitesh; Denys, Mark B

    2012-09-01

    Nanoparticles, when homogeneously dispersed in a base fluid, e.g. water, ethylene glycol etc. are commonly known as nanofluids. Nanofluids have gained attention in the scientific community for their enhanced thermal properties. One of the major problems in using nanofluids as a heat transfer medium for commercial applications is that, in most of the closed circuit industrial cooling processes, the cooling fluid has to be replaced after several cycles of cooling operation because of an increased presence of contaminants. If nanofluids were used as a coolant, it would be very hard to separate the nanoparticles from the waste fluid. The present work is aimed at the separation and recycling of nanoparticles from fluid waste by means of quick settling of titanium dioxide nanoparticles using silver nanoparticles along with ultrasonic treatment. It is observed that with increasing silver concentration and time of ultrasonication, the stability of the dispersion decreases. There is a value for both the silver concentration and ultrasonication time above which the settling time decreases drastically. PMID:22421063

  18. Additional Electrochemical Treatment Effects on the Switching Characteristics of Anodic Porous Alumina Resistive Switching Memory

    NASA Astrophysics Data System (ADS)

    Otsuka, Shintaro; Takeda, Ryouta; Furuya, Saeko; Shimizu, Tomohiro; Shingubara, Shouso; Iwata, Nobuyuki; Watanabe, Tadataka; Takano, Yoshiki; Takase, Kouichi

    2012-06-01

    We have investigated the current-voltage characteristics of a resistive switching memory (ReRAM), especially the reproducibility of the switching voltage between an insulating state and a metallic state. The poor reproducibility hinders the practical use of this memory. According to a filament model, the variation of the switching voltage may be understood in terms of the random choice of filaments with different conductivities and lengths at each switching. A limitation of the number of conductive paths is expected to lead to the suppression of the variation of switching voltage. In this study, two strategies for the limitation have been proposed using an anodic porous alumina (APA). The first is the reduction of the number of conductive paths by restriction of the contact area between the top electrodes and the insulator. The second is the lowering of the resistivity of the insulator, which makes it possible to grow filaments with the same characteristics by electrochemical treatments using a pulse-electroplating technique.

  19. Molecular identification of Cryptosporidium spp. and Giardia duodenalis in grazing horses from Xinjiang, China.

    PubMed

    Qi, Meng; Zhou, Huan; Wang, Haiyan; Wang, Rongjun; Xiao, Lihua; Arrowood, Michael J; Li, Junqiang; Zhang, Longxian

    2015-04-30

    A total of 262 fecal specimens collected from grazing horses at five locations in Xinjiang, China were examined by PCR for Cryptosporidium spp. and Giardia duodenalis. The Cryptosporidium and G. duodenalis infection rates were 2.7% and 1.5%, respectively. Seven Cryptosporidium-positive specimens were found in foals (16.3%), and four G. duodenalis-positive specimens were found in mares (2.5%). Sequence analyses of 18S rRNA and gp60 genes revealed that seven animals were positive for the subtype VIaA15G4 of Cryptosporidium horse genotype. G. duodenalis assemblages A and B were identified by molecular characterization of the 16S rRNA and tpi genes. This is the first report of Cryptosporidium horse genotype and G. duodenalis in grazing horses from China. PMID:25794943

  20. New pyrometallurgical process of EAF dust treatment with CaO addition

    NASA Astrophysics Data System (ADS)

    Chairaksa-Fujimoto, Romchat; Inoue, Yosuke; Umeda, Naoyoshi; Itoh, Satoshi; Nagasaka, Tetsuya

    2015-08-01

    The non-carbothermic zinc pyrometallurgical processing of electric arc furnace (EAF) dust was investigated on a laboratory scale. The main objective of this process was to convert highly stable zinc ferrite (ZnFe2O4), which accounts for more than half of total zinc in the EAF dust, into ZnO and Ca2Fe2O5 by CaO addition. The EAF dust was mixed with CaO powder in various ratios, pressed into pellets, and heated in a muffle furnace in air at temperatures ranging from 700 to 1100°C for a predetermined holding time. All ZnFe2O4 was transformed into ZnO and Ca2Fe2O5 at a minimum temperature of 900°C within 1 h when sufficient CaO to achieve a Ca/Fe molar ratio of 1.1 was added. However, at higher temperatures, excess CaO beyond the stoichiometric ratio was required because it was consumed by reactions leading to the formation of compounds other than ZnFe2O4. The evaporation of halides and heavy metals in the EAF dust was also studied. These components could be preferentially volatilized into the gas phase at 1100°C when CaO was added.

  1. Oxidative addition of hydrogen to bis(phosphine)platinum(0) complexes: an ab initio theroretical treatment

    SciTech Connect

    Noell, J.O.; Hay, P.J.

    1982-08-25

    Ab initio molecular orbital methods utilizing relativistic core potentials and correlated wave functions are employed to examine the oxidative addition reactions H/sub 2/ + Pt(PH/sub 3/)/sub 2/..-->..cis-Pt(PH/sub 3/)/sub 2/H/sub 2/ and H/sub 2/ + Pt(P(CH/sub 3/)/sub 3/)/sub 2/..-->..cis-Pt(P(CH/sub 3/)/sub 3/)/sub 2/H/sub 2/. For this symmetry-allowed process, an activation barrier of 17 kcal/mol and an exothermicity of 7 kcal/mol are calculated at the SCF level for the PH/sub 3/ liquid; similar values are obtained for the P(CH/sub 3/)/sub 3/ ligand. This implies a barrier of 24 kcal/mol for the reverse reductive elimination reaction. These values were not significantly altered in MC-SCF or CI calculations. This barrier is consistent with available data in the analogous process in six-coordinate complexes but is puzzling in light of the paucity of known four-coordinate cis dihydrides. The reaction is analyzed in terms of three phases: initial repulsion, partial transfer of charge from the platinum to the hydrogen, and final metal-hydrogen bond formation. The relative energies of the cis and trans isomers are also discussed.

  2. Indication of advanced orthokeratology as an additional treatment after refractive surgeries

    NASA Astrophysics Data System (ADS)

    Mitsui, Iwane; Yamada, Yoshida

    2005-04-01

    Ortho-K was indicated for twenty-three eyes of thirteen patients after refractive surgeries such as RK(1) ,PRK(2), and LASIK(3). The average of their Uncorrective Visual Acuity (UCVA) after surgeries was 20/30 or worse, and mean spherical equivalent (SE) was -2.42D. They were followed at least two years wearing of Advanced Ortho-K lenses during night. The following studies were examined on their auto-refraction, auto-keratometry, uncorrected and corrected visual acuity, intra-ocular pressure, corneal endothelium, corneal thickness, corneal curvature, and corneal shape for more than two years. 95% of the patients improved in UCVA up to 20/20 or better, 86% of them improved up to 20/15 or better, and 76% of them improved up to 20/10. The mean SEs improved to -1.20+/-1.02D during six months, - 1.03+/-0.83D during one year, and -0.73+/-0.64D during two years. Astigmatism also slightly decreased. Ophthalmologic examinations showed no abnormalities including flap formation, intra-ocular pressure, and endothelium. Among the refractive surgeries as well as RK and PRK, LASIK has been most popularly spread all over the world. However, patient's quality of vision is not always satisfied during and/or after refractive surgeries, because of several complications such as instability of flap formation, unexpected keratoectasia, diffuse lamellar keratitis, epithelial ingrowth, irregularity of corneal surface which caused myopia regression. In such cases, additional surgical procedures should not be indicated easily. However, Ortho-K is safe and effective enough to correct refractive errors still remained or re-appeared after refractive surgeries. It enables to restore the corneal irregularity to the ideal shape.

  3. Does the Addition of a Second Antipsychotic Drug Improve Clozapine Treatment?

    PubMed Central

    Barbui, Corrado; Signoretti, Alessandra; Mulè, Serena; Boso, Marianna; Cipriani, Andrea

    2009-01-01

    In patients with schizophrenia who do not have an optimal response to clozapine, it remains unclear if there is an evidence base to support a second antipsychotic in combination with clozapine. The present systematic review was therefore carried out to determine the efficacy of various clozapine combination strategies with antipsychotics. Relevant studies were located by searching the Cochrane Schizophrenia Group Trials Register, Medline, and Embase (up to November 2007). Only studies randomly allocating patients to clozapine plus another antipsychotic vs clozapine monotherapy were included. The search yielded 21 studies suitable for reanalysis. In 3 trials, clozapine was combined with a phenothiazine, in 8 trials with a benzamide, and in the remaining trials with risperidone. While the majority of randomized trials were not double blind, 6 studies were double-blind placebo-controlled trials. A total of 14 randomized open studies significantly favored clozapine combination strategy in terms of mean difference (random effect standardized mean difference [SMD] = −0.80, 95% confidence interval [CI] = −1.14 to −0.46); however, data extracted from 6 randomized double-blind studies did not show a statistically significant positive effect of this combination strategy in terms of mean difference (SMD = −0.12, 95% CI = −0.57 to 0.32). In terms of percentage of patients failing to show an improvement, a total of 10 randomized open studies significantly favored clozapine combination strategy (random effect relative risk [RR] = 0.64, 95% CI = 0.42 to 0.97), but data extracted from 6 randomized double-blind studies did not show a statistically significant positive effect of this combination strategy (RR = 0.91, 95% CI = 0.75 to 1.11). We conclude that the evidence base supporting a second antipsychotic in addition to clozapine in partially responsive patients with schizophrenia is weak. This weak evidence indicates modest to absent benefit. PMID:18436527

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

    PubMed

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

    2015-08-01

    Little is known of the prevalence of Cryptosporidium and Giardia in domestic cats in Western Australia and their potential role as zoonotic reservoirs for human infection. In the present study, a total of 345 faecal samples from four different sources were screened for the presence of Cryptosporidium and Giardia by PCR and genotyped by sequence analysis. Oocyst numbers and cyst numbers for Cryptosporidium and Giardia respectively were also determined using quantitative PCR assays. Cryptosporidium and Giardia were detected in 9.9% (95% CI 6.7-13.0) and 10.1% (95% CI 7.0-13.3) of cats in Western Australia respectively. Sequence analysis at the 18S rRNA locus identified five Cryptosporidium species/genotypes; C. felis (n = 8), C. muris (n = 1), C. ryanae (n = 1), Cryptosporidium rat genotype III (n = 5) and a novel genotype most closely related to Cryptosporidium rat genotype III in one isolate. This is the first report of C. ryanae and Cryptosporidium rat genotype III in cats. For Giardia, assemblage F the most commonly identified species, while only 1 assemblage sequence was detected. Since most human cases of cryptosporidiosis are caused by C. parvum and C. hominis and human cases of giardiasis are caused by G. duodenalis assemblage A and B, the domestic cats in the present study are likely to be of low zoonotic risk to pet owners in Perth. Risk analyses identified that elderly cats (more than 6 years) were more prone to Cryptosporidium and Giardia infections than kittens (less than 6 months) (P = 0.009). Clinical symptoms were not associated with the prevalence of Cryptosporidium and Giardia infections in cats. PMID:25959691

  5. Genetic Diversity of Cryptosporidium spp. within a Remote Population of Soay Sheep on St. Kilda Islands, Scotland

    PubMed Central

    Connelly, L.; Craig, B. H.; Jones, B.

    2013-01-01

    This is the first report to characterize the genotypes and subtypes of Cryptosporidium species infecting a geographically isolated population of feral Soay sheep (Ovis aries) on Hirta, St. Kilda, Scotland, during two distinct periods: (i) prior to a population crash and (ii) as host numbers increased. Cryptosporidium DNA was extracted by freeze-thawing of immunomagnetically separated (IMS) bead-oocyst complexes, and species were identified following nested-PCR-restriction fragment length polymorphism (RFLP)/PCR sequencing at two Cryptosporidium 18S rRNA loci. Two hundred fifty-five samples were analyzed, and the prevalent Cryptosporidium species in single infections were identified as C. hominis (11.4% of all samples tested), C. parvum (9%), C. xiaoi (12.5%), and C. ubiquitum (6.7%). Cryptosporidium parvum was also present with other Cryptosporidium species in 27.1% of all samples tested. Cryptosporidium parvum- and C. hominis-positive isolates were genotyped using two nested-PCR assays that amplify the Cryptosporidium glycoprotein 60 gene (GP60). GP60 gene analysis showed the presence of two Cryptosporidium genotypes, namely, C. parvum IIaA19G1R1 and C. hominis IbA10G2. This study reveals a higher diversity of Cryptosporidium species/genotypes than was previously expected. We suggest reasons for the high diversity of Cryptosporidium parasites within this isolated population and discuss the implications for our understanding of cryptosporidiosis. PMID:23354707

  6. The Effectiveness of Additional Treatment Modalities after the Failure of Recanalization by Thrombectomy Alone in Acute Vertebrobasilar Arterial Occlusion

    PubMed Central

    Kim, Seong Mook; Sohn, Sung-Il; Hong, Jeong-Ho; Chang, Hyuk-Won; Lee, Chang-Young

    2015-01-01

    Objective Acute vertebrobasilar artery occlusion (AVBAO) is a devastating disease with a high mortality rate. One of the most important factors affecting favorable clinical outcome is early recanalization. Mechanical thrombectomy is an emerging treatment strategy for achieving a high recanalization rates. However, thrombectomy alone can be insufficient to complete recanalization, especially for acute stroke involving large artery atheromatous disease. The purpose of this study is to investigate the safety and efficacy of mechanical thrombectomy in AVBAO. Methods Fourteen consecutive patients with AVBAO were treated with mechanical thrombectomy. Additional multimodal treatments were intra-arterial (IA) thrombolysis, balloon angioplasty, or permanent stent placement. Recanalization by thrombectomy alone and multimodal treatments were assessed by the Thrombolysis in Cerebral Infarction (TICI) score. Clinical outcome was determined using the National Institutes of Health Stroke Scale (NIHSS) at 7 days and the modified Rankin Scale (mRS) at 3 months. Results Thrombectomy alone and multimodal treatments were performed in 10 patients (71.4%) and 4 patients (28.6%), respectively. Successful recanalization (TICI 2b-3) was achieved in 11 (78.6%). Among these 11 patients, 3 (27.3%) underwent multimodal treatment due to underlying atherosclerotic stenosis. Ten (71.4%) of the 14 showed NIHSS score improvement of >10. Overall mortality was 3 (21.4%) of 14. Conclusion We suggest that mechanical thrombectomy is safe and effective for improving recanalization rates in AVBAO, with low complication rates. Also, in carefully selected patients after the failure of recanalization by thrombectomy alone, additional multimodal treatment such as IA thrombolysis, balloons, or stents can be needed to achieve successful recanalization. PMID:26713141

  7. Genotypes of Cryptosporidium spp. and Enterocytozoon bieneusi in Human Immunodeficiency Virus-Infected Patients in Lagos, Nigeria.

    PubMed

    Ojuromi, Oladele T; Duan, Liping; Izquierdo, Fernando; Fenoy, Soledad M; Oyibo, Wellington A; Del Aguila, Carmen; Ashafa, Anofi O T; Feng, Yaoyu; Xiao, Lihua

    2016-07-01

    Molecular characterization of Cryptosporidium spp. and Enterocytozoon bieneusi has improved our understanding of the transmission of both organisms in humans. In this study, to infer possible infection sources, Cryptosporidium spp. and E. bieneusi in fecal specimens from 90 HIV-infected patients attending antiretroviral clinics in Lagos, Nigeria were detected and genotyped by PCR and DNA sequencing. Cryptosporidium spp. and E. bieneusi were identified in four and five patients, respectively, including the occurrence of subtype IeA11T3G3 of Cryptosporidium hominis in two patients, subtype IIcA5G3k of Cryptosporidium parvum in one patient, and Type IV of E. bieneusi in four patients. Among the remaining positive patients, one had mixed infection of Cryptosporidium meleagridis and C. hominis and one had mixed E. bieneusi genotypes. These data highlight a possible difference in major transmission routes (anthroponotic vs. zoonotic) between Cryptosporidium spp. and E. bieneusi in HIV+ patients in the study area. PMID:26662459

  8. Identification of Cryptosporidium parvum Dihydrofolate Reductase Inhibitors by Complementation in Saccharomyces cerevisiae

    PubMed Central

    Brophy, Victoria Hertle; Vasquez, John; Nelson, Richard G.; Forney, John R.; Rosowsky, Andre; Sibley, Carol Hopkins

    2000-01-01

    There is a pressing need for drugs effective against the opportunistic protozoan pathogen Cryptosporidium parvum. Folate metabolic enzymes and enzymes of the thymidylate cycle, particularly dihydrofolate reductase (DHFR), have been widely exploited as chemotherapeutic targets. Although many DHFR inhibitors have been synthesized, only a few have been tested against C. parvum. To expedite and facilitate the discovery of effective anti-Cryptosporidium antifolates, we have developed a rapid and facile method to screen potential inhibitors of C. parvum DHFR using the model eukaryote, Saccharomyces cerevisiae. We expressed the DHFR genes of C. parvum, Plasmodium falciparum, Toxoplasma gondii, Pneumocystis carinii, and humans in the same DHFR-deficient yeast strain and observed that each heterologous enzyme complemented the yeast DHFR deficiency. In this work we describe our use of the complementation system to screen known DHFR inhibitors and our discovery of several compounds that inhibited the growth of yeast reliant on the C. parvum enzyme. These same compounds were also potent or selective inhibitors of the purified recombinant C. parvum DHFR enzyme. Six novel lipophilic DHFR inhibitors potently inhibited the growth of yeast expressing C. parvum DHFR. However, the inhibition was nonselective, as these compounds also strongly inhibited the growth of yeast dependent on the human enzyme. Conversely, the antibacterial DHFR inhibitor trimethoprim and two close structural analogs were highly selective, but weak, inhibitors of yeast complemented by the C. parvum enzyme. Future chemical refinement of the potent and selective lead compounds identified in this study may allow the design of an efficacious antifolate drug for the treatment of cryptosporidiosis. PMID:10722506

  9. Effect of two-stage coagulant addition on coagulation-ultrafiltration process for treatment of humic-rich water.

    PubMed

    Liu, Ting; Chen, Zhong-lin; Yu, Wen-zheng; Shen, Ji-min; Gregory, John

    2011-08-01

    A novel two-stage coagulant addition strategy applied in a coagulation-ultrafiltration (UF) process for treatment of humic-rich water at neutral pH was investigated in this study. When aluminum sulfate (alum) doses were set at a ratio of 3:1 added during rapid mix stage and half way through flocculation stage, the integrated process of two-stage alum addition achieved almost the same organic matter removal as that of conventional one-stage alum addition at the same overall dose. Whereas membrane fouling could be effectively mitigated by the two-stage addition exhibited by trans-membrane pressure (TMP) developments. The TMP developments were found to be primarily attributed to external fouling on membrane surface, which was closely associated with floc characteristics. The results of jar tests indicated that the average size of flocs formed in two-stage addition mode roughly reached one half larger than that in one-stage addition mode, which implied a beneficial effect on membrane fouling reduction. Moreover, the flocs with more irregular structure and lower effective density resulted from the two-stage alum addition, which caused higher porosity of cake layer formed by such flocs on membrane surface. Microscopic observations of membrane surface demonstrated that internal fouling in membrane pores could be also remarkably limited by two-stage alum addition. It is likely that the freshly formed hydroxide precipitates were distinct in surface characteristics from the aged precipitates due to formation of more active groups or adsorption of more labile aluminum species. Consequently, the flocs could further connect and aggregate to contribute to preferable properties for filtration performance of the coagulation-UF process. As a simple and efficient approach, two-stage coagulant addition strategy could have great practical significance in coagulation-membrane processes. PMID:21704354

  10. Intestinal Parasite Profile in the Stool of HIV Positive Patients in relation to Immune Status and Comparison of Various Diagnostic Techniques with Special Reference to Cryptosporidium at a Tertiary Care Hospital in South India

    PubMed Central

    Moorkoth, Anitha Puduvail; Mathew, Sheela

    2016-01-01

    Acquired immunodeficiency syndrome and related opportunistic infections are a significant cause of morbidity and mortality in susceptible population. This study aims to negate the paucity of data regarding the relation between CD4 levels, prevalence of enteric parasites, and the outcome of treatment with HAART (highly active antiretroviral therapy) and Cotrimoxazole in Kerala, India. Multiple stool samples from 200 patients in a cross-sectional study were subjected to microscopy and Cryptosporidium stool antigen ELISA. Parasites were identified in 18 samples (9%). Cystoisospora and Cryptosporidium spp. were seen in 9 cases (4.5%) and 5 cases (2.5%), respectively. Microsporidium spores and Chilomastix mesnili cysts were identified in 1 case each (0.5% each). Seven cases of Cystoisospora diarrhoea recovered after treatment with Cotrimoxazole. Diarrhoea due to Cryptosporidium spp. in all 5 cases subsided after immune reconstitution with HAART. This study concludes that a positive association was seen between low CD4 count (<200 cells/μL) and overall parasite positivity (P value < 0.01). ELISA is a more sensitive modality for the diagnosis of Cryptosporidium diarrhoea. Chilomastix mesnili, generally considered a nonpathogen, may be a cause of diarrhoeal disease in AIDS. Immune reconstitution and Cotrimoxazole prophylaxis remain to be the best therapeutic approach in AIDS-related diarrhoea. PMID:27493988

  11. Electrical impedance sensor for quantitative monitoring of infection processes on HCT-8 cells by the waterborne parasite Cryptosporidium.

    PubMed

    Dibao-Dina, Alfred; Follet, Jérôme; Ibrahim, Mouhamad; Vlandas, Alexis; Senez, Vincent

    2015-04-15

    Cryptosporidium is the main origin of worldwide waterborne epidemic outbreaks caused by protozoan parasites. Its resilience to water chemical treatments and the absence of therapy led to consider it as a reference pathogen to assess water quality and as a possible bioterrorism agent. We here show that an electrical impedance-based device is able to get insights on Cryptosporidium development on a cell culture and to quantify sample infectivity. HCT-8 cells were grown to confluency on Interdigitated Microelectrode Arrays (IMA's) during 76h and then infected by Cryptosporidium parvum during 60h. The impedimetric response was measured at frequencies ranging from 100Hz to 1MHz and a 7min sampling period. As the infection progresses the impedance signal shows a reproducible distinct succession of peaks at 12h post infection (PI), 23h PI and 31h PI and local minima at 9h PI, 19h PI and 28h PI. An equivalent circuit modeling-based approach indicates that these features are mostly originated from paracellular pathway modifications due to host-parasite interactions. Furthermore, our data present for the first time a real-time monitoring of early parasitic stage development with alternating zoite and meront predominances, observed respectively at peaks and local minima in the impedimetric signal. Finally, by quantifying the magnitude of the impedimetric response, we demonstrate this device can also be used as an infectivity sensor as early as 12h PI thus being at least 6 times faster than other state of the art techniques. PMID:25460884

  12. Enhanced performance of a submerged membrane bioreactor with powdered activated carbon addition for municipal secondary effluent treatment.

    PubMed

    Lin, Hongjun; Wang, Fangyuan; Ding, Linxian; Hong, Huachang; Chen, Jianrong; Lu, Xiaofeng

    2011-09-15

    The aim of this study was to investigate the feasibility of PAC-MBR process treating municipal secondary effluent. Two laboratory-scale submerged MBRs (SMBR) with and without PAC addition were continuously operated in parallel for secondary effluent treatment. Approximately 63%TOC, 95% NH(4)(+)-N and 98% turbidity in secondary effluent were removed by the PAC-MBR process. Most organics in the secondary effluent were found to be low molecular weight (MW) substances, which could be retained in the reactor and then removed to some extent by using PAC-MBR process. Parallel experiments showed that the addition of PAC significantly increased organic removal and responsible for the largest fraction of organic removal. Membrane fouling analysis showed the enhanced membrane performance in terms of sustainable operational time and filtration resistances by PAC addition. Based on these results, the PAC-MBR process was considered as an attractive option for the reduction of pollutants in secondary effluent. PMID:21794980

  13. Prevalence and age-related infection of Cryptosporidium suis, C. muris and Cryptosporidium pig genotype II in pigs on a farm complex in the Czech Republic.

    PubMed

    Kvác, Martin; Hanzlíková, Dagmar; Sak, Bohumil; Kvetonová, Dana

    2009-03-23

    A total of 413 pig faecal samples were collected from pre-weaners (119), starters (131), pre-growers (123) and sows (40) from a farm with a closed breeding system segmented into two breeding complexes and a growing complex in the region of Vysocina, Czech Republic and screened for the presence of Cryptosporidium using staining methods and genotyping (SSU rRNA). Cryptosporidium oocysts were detected by microscopy in the faeces of 21.1% of the samples (87/413). Sequence analyses and RFLP identified C. suis in 44, Cryptosporidium pig genotype II in 23 and C. muris in 2 samples. No mixed infections were found. Pigs under 7 weeks of age were infected with C. suis only. Cryptosporidium pig genotype II was found in animals from 7 weeks of age. No relationship was found between diarrhoea and any Cryptosporidium infection in any of the different age groups (P<0.05). The pre-weaned pigs shed significantly more Cryptosporidium oocysts than older pigs and it was associated with C. suis infection. PMID:19091471

  14. Improving the efficiency of polymer solar cells based on furan-flanked diketopyrrolopyrrole copolymer via solvent additive and methanol treatment

    NASA Astrophysics Data System (ADS)

    Zhou, Weilong; Chen, Huajie; Lv, Junjie; Chen, Youchun; Zhang, Weifeng; Yu, Gui; Li, Fenghong

    2015-09-01

    We present a furan-flanked DPP copolymer, poly{3,6-difuran-2-yl-2,5-di(2-octyldodecyl)-pyrrolo [3,4-c]pyrrole-1,4-dione-altthienylenevinylene} (PDVF-8), and highlight the improvement in the power conversion efficiency (PCE) of polymer solar cells (PSCs) based on the PDVF-8 as an electron donor via solvent additive and methanol treatment. When 3 vol% 1,8-diiodooctane (DIO) or 1-chloronaphthalene (CN) were used as a solvent additive to the PDVF-8:PC71BM solution in chloroform (CF), the PCE can increase from 0.79% to 3.73% or 4.26%. Methanol treatment (MT) can further enhance the PCE to 4.03% (DIO) and 4.69% (CN). The effect of the solvent additives (DIO and CN) and MT on the phase separation of the PDVF-8:PC71BM thin film has been investigated in detail using atomic force microscopy, transmission electron microscopy (TEM), TEM-energy dispersive spectroscopy and X-ray photoemission spectroscopy depth profiling.We present a furan-flanked DPP copolymer, poly{3,6-difuran-2-yl-2,5-di(2-octyldodecyl)-pyrrolo [3,4-c]pyrrole-1,4-dione-altthienylenevinylene} (PDVF-8), and highlight the improvement in the power conversion efficiency (PCE) of polymer solar cells (PSCs) based on the PDVF-8 as an electron donor via solvent additive and methanol treatment. When 3 vol% 1,8-diiodooctane (DIO) or 1-chloronaphthalene (CN) were used as a solvent additive to the PDVF-8:PC71BM solution in chloroform (CF), the PCE can increase from 0.79% to 3.73% or 4.26%. Methanol treatment (MT) can further enhance the PCE to 4.03% (DIO) and 4.69% (CN). The effect of the solvent additives (DIO and CN) and MT on the phase separation of the PDVF-8:PC71BM thin film has been investigated in detail using atomic force microscopy, transmission electron microscopy (TEM), TEM-energy dispersive spectroscopy and X-ray photoemission spectroscopy depth profiling. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr04393j

  15. Effects of plasmochemical treatments and cerium additions on the structural characteristics and activity of copper catalyst particles in isopropanol dehydrogenation

    NASA Astrophysics Data System (ADS)

    Platonov, E. A.; Lobanov, N. N.; Galimova, N. A.; Protasova, I. A.; Yagodovskii, V. D.

    2012-08-01

    The effect of the treatment of the 5 wt % Cu/SiO2 (I) and (5 wt % Cu + 0.5 wt % Ce)/SiO2 (II) catalysts with glow discharge plasma in O2, H2, and Ar on their structural characteristics was studied by X-ray phase analysis; the influence of cerium additions and plasmochemical treatments on the catalyst activity in isopropanol dehydrogenation was also investigated. Under the plasmochemical treatment, the diameters of Cu particles in catalyst I nearly doubled and microstresses in the metal particles also changed. Catalyst II was X-ray amorphous both before and after plasmochemical treatments. The activity of I after plasmochemical treatment increased because of the increase in the number of centers and changes in their composition. Growth of the activity of I compared with the activity of II was explained by the formation of new catalytic centers due to positive charging of the Ce+α adatom on the surface of the copper particle.

  16. Survival of Infectious Cryptosporidium parvum Oocysts in Seawater and Eastern Oysters (Crassostrea virginica) in the Chesapeake Bay

    PubMed Central

    Fayer, Ronald; Graczyk, Thaddeus K.; Lewis, Earl J.; Trout, James M.; Farley, C. Austin

    1998-01-01

    Oocysts of Cryptosporidium parvum placed in artificial seawater at salinities of 10, 20, and 30 ppt at 10°C and at 10 ppt at 20°C were infectious after 12 weeks. Those placed in seawater at 20 ppt and 30 ppt at 20°C were infectious for 8 and 4 weeks, respectively. These findings suggested that oocysts could survive in estuarine waters long enough to be removed by filter feeders such as oysters. Thereafter, 30 Eastern oysters, Crassostrea virginica, were collected with a dredge or with hand tongs at each of six sites within Maryland tributaries of the Chesapeake Bay in May and June and in August and September of 1997. Hemocytes and gill washings from all oysters were examined for the presence of Cryptosporidium oocysts and Giardia cysts by immunofluorescence microscopy utilizing a commercially available kit containing fluorescein isothiocyanate-conjugated monoclonal antibodies. Giardia was not detected by this method from any of the 360 oysters examined. Presumptive identification of Cryptosporidium oocysts was made in either hemocytes or gill washings of oysters from all six sites both times that surveys were conducted. In addition, during August and September, for each of the six sites, hemocytes from the 30 oysters were pooled and gill washings from the oysters were pooled. Each pool was delivered by gastric intubation to a litter of neonatal mice to produce a bioassay for oocyst infectivity. Intestinal tissue from two of three mice that received gill washings from oysters collected at a site near a large cattle farm and shoreline homes with septic tanks was positive for developmental stages of C. parvum. These findings demonstrate for the first time that oysters in natural waters harbor infectious C. parvum oocysts and can serve as mechanical vectors of this pathogen. PMID:9501446

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

    PubMed Central

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

    2016-01-01

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

  18. Effect of water treatment additives on lime softening residual trace chemical composition--implications for disposal and reuse.

    PubMed

    Cheng, Weizhi; Roessler, Justin; Blaisi, Nawaf I; Townsend, Timothy G

    2014-12-01

    Drinking water treatment residues (WTR) offer potential benefits when recycled through land application. The current guidance in Florida, US allows for unrestricted land application of lime softening WTR; alum and ferric WTR require additional evaluation of total and leachable concentrations of select trace metals prior to land application. In some cases a mixed WTR is produced when lime softening is accompanied by the addition of a coagulant or other treatment chemical; applicability of the current guidance is unclear. The objective of this research was to characterize the total and leachable chemical content of WTR from Florida facilities that utilize multiple treatment chemicals. Lime and mixed lime WTR samples were collected from 18 water treatment facilities in Florida. Total and leachable concentrations of the WTR were measured. To assess the potential for disposal of mixed WTR as clean fill below the water table, leaching tests were conducted at multiple liquid to solid ratios and under reducing conditions. The results were compared to risk-based soil and groundwater contamination thresholds. Total metal concentrations of WTR were found to be below Florida soil contaminant thresholds with Fe found in the highest abundance at a concentration of 3600 mg/kg-dry. Aluminum was the only element that exceeded the Florida groundwater contaminant thresholds using SPLP (95% UCL = 0.23 mg/L; risk threshold = 0.2 mg/L). Tests under reducing conditions showed elevated concentrations of Fe and Mn, ranging from 1 to 3 orders of magnitude higher than SPLP leachates. Mixed lime WTR concentrations (total and leachable) were lower than the ferric and alum WTR concentrations, supporting that mixed WTR are appropriately represented as lime WTR. Testing of WTR under reducing conditions demonstrated the potential for release of certain trace metals (Fe, Al, Mn) above applicable regulatory thresholds; additional evaluation is needed to assess management options where

  19. Radiological, physical, and chemical characterization of additional alpha contaminated and mixed low-level waste for treatment at the advanced mixed waste treatment project

    SciTech Connect

    Hutchinson, D.P.

    1995-07-01

    This document provides physical, chemical, and radiological descriptive information for a portion of mixed waste that is potentially available for private sector treatment. The format and contents are designed to provide treatment vendors with preliminary information on the characteristics and properties for additional candidate portions of the Idaho National Engineering Laboratory (INEL) and offsite mixed wastes not covered in the two previous characterization reports for the INEL-stored low-level alpha-contaminated and transuranic wastes. This report defines the waste, provides background information, briefly reviews the requirements of the Federal Facility Compliance Act (P.L. 102-386), and relates the Site Treatment Plans developed under the Federal Facility Compliance Act to the waste streams described herein. Each waste is summarized in a Waste Profile Sheet with text, charts, and tables of waste descriptive information for a particular waste stream. A discussion of the availability and uncertainty of data for these waste streams precedes the characterization descriptions.

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

    PubMed

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

    2016-05-01

    The genus Cryptosporidium, which is an obligate intracellular parasite, infects various vertebrates and causes a diarrheal disease known as cryptosporidiosis. Bats are naturally infected with zoonotic pathogens; thus, they are potential reservoirs of parasites. We investigated the species and genotype distribution as well as prevalence of Cryptosporidium and Eimeria in Philippine bats. We captured and examined 45 bats; four were positive for Cryptosporidium spp. and seven were positive for Eimeria spp. We detected Cryptosporidium bat genotype II from Ptenochirus jagori. Three other Cryptosporidium sequences, detected from Rhinolophus inops, Cynopterus brachyotis, and Eonycteris spelaea, could not be classified as any known species or genotype; we therefore propose the novel genotype Cryptosporidium bat genotypes V, VI, and VII. Bat genotype V is associated with human cryptosporidiosis clade, and therefore, this genotype may be transmissible to humans. Among the Eimeria sequences, BE3 detected from Scotophilus kuhlii was classified with known bat and rodent clades; however, other sequences detected from C. brachyotis, E. spelaea, Rousettus amplexicaudatus, and R. inops could not be classified with known Eimeria species. These isolates might represent a new genotype. Our findings demonstrate that the bats of the Philippines represent a reservoir of multiple Cryptosporidium and Eimeria spp. PMID:26833326

  1. Identification and differentiation of Cryptosporidium species by capillary electrophoresis single-strand conformation polymorphism.

    PubMed

    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. PMID:21087296

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

    PubMed

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

    2016-08-01

    It has been demonstrated that the allergic response can be ameliorated by the administration of pathogen derivatives that activate Toll-like receptors and induce a Th1-type immune response (IR). Cryptosporidium is a parasite that promotes an IR via Toll-like receptors and elicits the production of Th1-type cytokines, which limit cryptosporidiosis. The aim of this study was to investigate allergy-related immune markers in children naturally infected with Cryptosporidium. In a cross-sectional study, 49 children with or without clinical diagnosis of allergies, oocysts of Cryptosporidium spp. in the faeces were screened microscopically. We microscopically screened for leucocytes, examined T and B cells for allergy-related activation markers using flow cytometry and evaluated serum for total IgE using chemiluminescence. Children with allergies and Cryptosporidium in the faeces had significantly lower levels of total IgE, B cells, CD19(+) CD23(+) and CD19(+) CD124(+) cells as well as a greater percentage of interferon-gamma (IFN-γ(+) ) and IL-4(+) CD4(+) cells than children with allergies without Cryptosporidium. This is the first description of the modulation of the IR in children with allergic diseases in the setting of natural Cryptosporidium infection. Our findings suggest the involvement of CD4(+) cells producing IL-4 and IFN-γ in the IR to Cryptosporidium in naturally infected children. PMID:27150641

  3. Molecular detection and characterization of Cryptosporidium spp. among breeding cattery cats in Japan.

    PubMed

    Ito, Yoichi; Itoh, Naoyuki; Kimura, Yuya; Kanai, Kazutaka

    2016-05-01

    Cryptosporidium spp. are pathogenic protozoan that can cause gastrointestinal illness in mammalian hosts. As a result of the close contact between humans and cats, there is concern regarding the potential zoonotic transmission of Cryptosporidium spp. from infected cats; however, few data have been reported regarding the prevalence of this pathogen among cats. Here, we report the prevalence of Cryptosporidium spp. among breeding cattery cats in Japan. A total of 286 fresh fecal samples were collected from breeding cattery cats at seven facilities located across Japan. A nested polymerase chain reaction (PCR) assay targeting the 18S rRNA gene was employed for the detection of Cryptosporidium spp. Four cats (1.4 %), from two catteries, were positive for Cryptosporidium spp. Age and fecal condition were not significantly associated with prevalence. The four positive samples displayed 99-100 % sequence similarity to Cryptosporidium felis sequences. Our findings indicated that the prevalence of Cryptosporidium spp. was low among breeding cattery cats in Japan, and therefore the risk of zoonotic transmission to humans was also likely to be low. PMID:26960959

  4. Identity and public health potential of Cryptosporidium spp. in water buffalo calves in Egypt.

    PubMed

    Amer, Said; Zidan, Shereif; Feng, Yaoyu; Adamu, Haileeyesus; Li, Na; Xiao, Lihua

    2013-01-16

    Little is known about the diversity and public health significance of Cryptosporidium species in water buffaloes. In this study, we examined the distribution of Cryptosporidium spp. in water buffalo calves in Egypt. Rectal fecal specimens from 179 calves and 359 adults were screened microscopically for Cryptosporidium oocysts using modified Ziehl-Neelsen stain. Cryptosporidium spp. in 17 microscopy-positive specimens from calves were genotyped by DNA sequence analysis of the small-subunit rRNA gene, and Cryptosporidium parvum was subtyped by sequence analysis of the 60 kDa glycoprotein gene. Cryptosporidium ryanae was found in 10 specimens and C. parvum in 7 specimens, with the former belonging to the newly identified C. ryanae buffalo variant and the latter belonging to the subtypes IIdA20G1 (in 5 specimens) and IIaA15G1R1 (in 2 specimens). The prevailing occurrence of C. ryanae and the subtype family IId of C. parvum and the absence of C. bovis and C. andersoni represent some features of Cryptosporidium transmission in water buffaloes in Egypt. PMID:22963712

  5. Novel anti-Cryptosporidium activity of known drugs identified by high-throughput screening against parasite fatty acyl-CoA binding protein (ACBP)

    PubMed Central

    Fritzler, Jason M.; Zhu, Guan

    2012-01-01

    Background Cryptosporidium parvum causes an opportunistic infection in AIDS patients, and no effective treatments are yet available. This parasite possesses a single fatty acyl-CoA binding protein (CpACBP1) that is localized to the unique parasitophorous vacuole membrane (PVM). The major goal of this study was to identify inhibitors from known drugs against CpACBP1 as potential new anti-Cryptosporidium agents. Methods A fluorescence assay was developed to detect CpACBP1 activity and to identify inhibitors by screening known drugs. Efficacies of top CpACBP1 inhibitors against Cryptosporidium growth in vitro were evaluated using a quantitative RT–PCR assay. Results Nitrobenzoxadiazole-labelled palmitoyl-CoA significantly increased the fluorescent emission upon binding to CpACBP1 (excitation/emission 460/538 nm), which was quantified to determine the CpACBP1 activity and binding kinetics. The fluorescence assay was used to screen a collection of 1040 compounds containing mostly known drugs, and identified the 28 most active compounds that could inhibit CpACBP1 activity with sub-micromolar IC50 values. Among them, four compounds displayed efficacies against parasite growth in vitro with low micromolar IC50 values. The effective compounds were broxyquinoline (IC50 64.9 μM), cloxyquin (IC50 25.1 μM), cloxacillin sodium (IC50 36.2 μM) and sodium dehydrocholate (IC50 53.2 μM). Conclusions The fluorescence ACBP assay can be effectively used to screen known drugs or other compound libraries. Novel anti-Cryptosporidium activity was observed in four top CpACBP1 inhibitors, which may be further investigated for their potential to be repurposed to treat cryptosporidiosis and to serve as leads for drug development. PMID:22167242

  6. Incorporation of exogenous uracil by Cryptosporidium parvum in vitro.

    PubMed Central

    Upton, S J; Tilley, M; Mitschler, R R; Oppert, B S

    1991-01-01

    Oocysts of Cryptosporidium parvum were used to infect Madin-Darby bovine kidney cells. Cultures were incubated in a reduced-oxygen atmosphere in candle jars or in a 5% CO2-95% air atmosphere. At 72 h, parasites were quantitated microscopically and found to be enhanced 5.5-fold in the reduced-oxygen atmosphere. Using candle jars, we then determined that C. parvum was amenable to [3H]uracil incorporation assays and easily quantitated with this method. Images PMID:2056042

  7. Cystamine and Intrabody Co-treatment Confers Additional Benefits in a Fly Model of Huntington’s Disease

    PubMed Central

    Bortvedt, S. F.; McLear, J. A.; Messer, A.; Ahern-Rindell, A. J.; Wolfgang, W. J.

    2010-01-01

    Huntington’s disease (HD) is a lethal, neurodegenerative disorder caused by expansion of the polyglutamine repeat in the Huntingtin gene (HTT), leading to mutant protein misfolding, aggregation, and neuronal death. Feeding a Drosophila HD-model cystamine, or expressing a transgene encoding the anti-htt intracellular antibody (intrabody) C4-scFv in the nervous system, demonstrated therapeutic potential, but suppression of pathology was incomplete. We hypothesized that a combinatorial approach entailing drug and intrabody administration could enhance rescue of HD pathology in flies and that timing of treatment would affect outcomes. Feeding cystamine to adult HD flies expressing the intrabody resulted in a significant, additional rescue of photoreceptor neurodegeneration, but no additional benefit in longevity. Feeding cystamine during both larval and adult stages produced the converse result: longevity was significantly improved, but increased photoreceptor survival was not. We conclude that cystamine-intrabody combination therapies can be effective, reducing neurodegeneration and prolonging survival, depending on administration protocols. PMID:20399860

  8. Additional Reserve Recovery Using New Polymer Treatment on High Water Oil Ratio Wells in Alameda Field, Kingman County, Kansas

    SciTech Connect

    James Spillane

    2005-10-01

    The Chemical Flooding process, like a polymer treatment, as a tertiary (enhanced) oil recovery process can be a very good solution based on the condition of this field and its low cost compared to the drilling of new wells. It is an improved water flooding method in which high molecular-weight (macro-size molecules) and water-soluble polymers are added to the injection water to improve the mobility ratio by enhancing the viscosity of the water and by reducing permeability in invaded zones during the process. In other words, it can improve the sweep efficiency by reducing the water mobility. This polymer treatment can be performed on the same active oil producer well rather than on an injector well in the existence of strong water drive in the formation. Some parameters must be considered before any polymer job is performed such as: formation temperature, permeability, oil gravity and viscosity, location and formation thickness of the well, amount of remaining recoverable oil, fluid levels, well productivity, water oil ratio (WOR) and existence of water drive. This improved oil recovery technique has been used widely and has significant potential to extend reservoir life by increasing the oil production and decreasing the water cut. This new technology has the greatest potential in reservoirs that are moderately heterogeneous, contain moderately viscous oils, and have adverse water-oil mobility ratios. For example, many wells in Kansas's Arbuckle formation had similar treatments and we have seen very effective results. In addition, there were previous polymer treatments conducted by Texaco in Alameda Field on a number of wells throughout the Viola-Simpson formation in the early 70's. Most of the treatments proved to be very successful.

  9. Binding mode of inhibitors and Cryptosporidium parvum IMP dehydrogenase: A combined ligand- and receptor-based study.

    PubMed

    Li, R-J; Wang, Y-L; Wang, Q-H; Huang, W-X; Wang, J; Cheng, M-S

    2015-01-01

    A combined ligand- and target-based approach was used to analyse the interaction models of Cryptosporidium parvum inosine 5'-monophosphate dehydrogenase (CpIMPDH) with selective inhibitors. First, a ligand-based pharmacophore model was generated from 20 NAD(+) competitive CpIMPDH inhibitors with the HipHop module. The characteristic of the NAD(+) binding site of CpIMPDH was then described, and the binding modes of the representative inhibitors were studied by molecular docking. The combination of the pharmacophore model and the docking results allowed us to evaluate the pharmacophore features and structural information of the NAD(+) binding site of CpIMPDH. This research supports the proposal of an interaction model inside the NAD(+) binding site of CpIMPDH, consisting of four key interaction points: two hydrophobic-aromatic groups, a hydrophobic-aliphatic group and a hydrogen bond donor. This study also provides guidance for the design of more potent CpIMPDH inhibitors for the treatment of Cryptosporidium infections. PMID:25978645

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

    PubMed

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

    2012-03-01

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

  11. CRYPTOSPORIDIUM: THE MILWAUKEE EXPERIENCE AND RELEVANT RESEARCH

    EPA Science Inventory

    The Surface Water Treatment Rule (SWTR) was mandated on June 29, 1989 by the U.S. Environmental Protection Agency (USEPA). his Rule requires all water utilities that use surface waters for their source water for drinking water to filter the water before sending it out into the di...

  12. Addition of home-based cognitive retraining to treatment as usual in first episode schizophrenia patients: A randomized controlled study

    PubMed Central

    Hegde, Shantala; Rao, Shobini L.; Raguram, Ahalya; Gangadhar, Bangalore N.

    2012-01-01

    Objective: We examined the effectiveness of a 2-month-long home-based cognitive retraining program together with treatment as usual (TAU; psychoeducation and drug therapy) on neuropsychological functions, psychopathology, and global functioning in patients with first episode schizophrenia (FES) as well as on psychological health and perception of level of family distress in their caregivers. Materials and Methods: Forty-five FES patients were randomly assigned to either treatment group receiving home-based cognitive retraining along with TAU (n=22) or to control group receiving TAU alone (n=23). Patients and caregivers received psychoeducation. Patients and one of their caregivers were assessed for the above parameters at baseline, post-assessment (2 months) and at 6-months follow-up assessment. Results: Of the 45 patients recruited, 12 in the treatment group and 11 in the control group completed post-intervention and follow-up assessments. Addition of home-based cognitive retraining along with TAU led to significant improvement in neuropsychological functions of divided attention, concept formation and set-shifting ability, and planning. Effect sizes were large, although the sample size was small. Conclusions: Home-based cognitive retraining program has shown promise. However, further studies examining this program on a larger cohort with rigorous design involving independent raters are suggested. PMID:22556432

  13. A new heterogeneous family of telomerically encoded Cryptosporidium proteins

    PubMed Central

    Bouzid, Maha; Hunter, Paul R; McDonald, Vincent; Elwin, Kristin; Chalmers, Rachel M; Tyler, Kevin M

    2013-01-01

    Cryptosporidiosis is predominantly caused by two closely related species of protozoan parasites the zoonotic Cryptosporidium parvum and anthroponotic Cryptosporidium hominis which diverge phenotypically in respect to host range and virulence. Using comparative genomics we identified two genes displaying overt heterogeneity between species. Although initial work suggested both were species specific, Cops-1 for C. parvum and Chos-1 for C. hominis, subsequent study identified an abridged ortholog of Cops-1 in C. hominis. Cops-1 and Chos-1 showed limited, but significant, similarity to each other and share common features: (i) telomeric location: Cops-1 is the last gene on chromosome 2, whilst Chos-1 is the first gene on chromosome 5, (ii) encode circa 50-kDa secreted proteins with isoelectric points above 10, (iii) are serine rich, and (iv) contain internal nucleotide repeats. Importantly, Cops-1 sequence contains specific SNPs with good discriminatory power useful epidemiologically. C. parvum-infected patient sera recognized a 50-kDa protein in antigen preparations of C. parvum but not C. hominis, consistent with Cops-1 being antigenic for patients. Interestingly, anti-Cops-1 monoclonal antibody (9E1) stained oocyst content and sporozoite surface of C. parvum only. This study provides a new example of protozoan telomeres as rapidly evolving contingency loci encoding putative virulence factors. PMID:23467513

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

    PubMed

    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. PMID:25066778

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

    PubMed Central

    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

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

    NASA Astrophysics Data System (ADS)

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

    2004-05-01

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

  17. Effects of anodizing parameters and heat treatment on nanotopographical features, bioactivity, and cell culture response of additively manufactured porous titanium.

    PubMed

    Amin Yavari, S; Chai, Y C; Böttger, A J; Wauthle, R; Schrooten, J; Weinans, H; Zadpoor, A A

    2015-06-01

    Anodizing could be used for bio-functionalization of the surfaces of titanium alloys. In this study, we use anodizing for creating nanotubes on the surface of porous titanium alloy bone substitutes manufactured using selective laser melting. Different sets of anodizing parameters (voltage: 10 or 20V anodizing time: 30min to 3h) are used for anodizing porous titanium structures that were later heat treated at 500°C. The nanotopographical features are examined using electron microscopy while the bioactivity of anodized surfaces is measured using immersion tests in the simulated body fluid (SBF). Moreover, the effects of anodizing and heat treatment on the performance of one representative anodized porous titanium structures are evaluated using in vitro cell culture assays using human periosteum-derived cells (hPDCs). It has been shown that while anodizing with different anodizing parameters results in very different nanotopographical features, i.e. nanotubes in the range of 20 to 55nm, anodized surfaces have limited apatite-forming ability regardless of the applied anodizing parameters. The results of in vitro cell culture show that both anodizing, and thus generation of regular nanotopographical feature, and heat treatment improve the cell culture response of porous titanium. In particular, cell proliferation measured using metabolic activity and DNA content was improved for anodized and heat treated as well as for anodized but not heat-treated specimens. Heat treatment additionally improved the cell attachment of porous titanium surfaces and upregulated expression of osteogenic markers. Anodized but not heat-treated specimens showed some limited signs of upregulated expression of osteogenic markers. In conclusion, while varying the anodizing parameters creates different nanotube structure, it does not improve apatite-forming ability of porous titanium. However, both anodizing and heat treatment at 500°C improve the cell culture response of porous titanium. PMID

  18. Perturbation of the intestinal microbiota of mice infected with Cryptosporidium parvum.

    PubMed

    Ras, Refaat; Huynh, Kevin; Desoky, Enas; Badawy, Ahmed; Widmer, Giovanni

    2015-07-01

    Understanding the interaction between the intestinal microbiota (microbiome) and enteric pathogens is of interest in the development of alternative treatments that do not rely on chemotherapy and do not lead to drug resistance. We undertook research in a rodent model of cryptosporidiosis to assess whether the bacterial gut microbiota is impacted by infection with the protozoan pathogen Cryptosporidium parvum. The profile of the faecal bacterial microbiota in infected and uninfected animals was compared using 16S amplicon sequencing. In four independent experiments, the intestinal microbiota of infected mice differed from that of uninfected animals, regardless of the C. parvum isolate used to infect mice. The use of replicated treatment groups demonstrated that microbiota divergence between treatments was driven by the infection and did not result from spontaneous changes in the intestinal ecosystem unrelated to the infection. Microbiota perturbation induced by C. parvum appeared to be reversible, as we observed a tendency for the phylogenetic distance between infected and uninfected mice to diminish after mice cleared the infection. As mice infected with C. parvum do not develop diarrhoea, these observations indicate that microbiota perturbation results from other mechanisms than an accelerated movement of gut content. PMID:25913477

  19. Commercial Assay for Detection of Giardia lamblia and Cryptosporidium parvum Antigens in Human Fecal Specimens by Rapid Solid-Phase Qualitative Immunochromatography

    PubMed Central

    Garcia, Lynne S.; Shimizu, Robyn Y.; Novak, Susan; Carroll, Marilyn; Chan, Frank

    2003-01-01

    The ImmunoCard STAT! Cryptosporidium/Giardia rapid assay (Meridian Bioscience, Inc.) is a solid-phase qualitative immunochromatographic assay that detects and distinguishes between Giardia lamblia and Cryptosporidium parvum in aqueous extracts of human fecal specimens (fresh, frozen, unfixed, or fixed in 5 or 10% formalin or sodium acetate-acetic acid-formalin). By using specific antibodies, antigens specific for these organisms are isolated and immobilized on a substrate. After the addition of appropriate reagents, a positive test is detected visually by the presence of a gray-black color bar (regardless of the intensity) next to the organism name printed on the test device. A control is included in the device. Steps include tube preparation (buffer, patient specimen, conjugates A and B), testing (addition of sample onto the test device), and visual reading (total time, 12 min). Test performance was evaluated with known positive and negative stool specimens (170 specimens positive for Giardia and 231 specimens negative for Giardia) (85 specimens positive for Cryptosporidium and 316 specimens negative for Cryptosporidium); they were tested with trichrome, iron-hematoxylin, or modified acid-fast stains or the Meridian Bioscience, Inc., Giardia/Cryptosporidium Merifluor combination reagent; specimens with discrepant results were retested by using the Merifluor combination reagent. On the basis of the results of the reference methods, the sensitivities, specificities, and positive and negative predictive values were as follows: for G. lamblia, 93.5, 100, 100, and 95.5%, respectively; for C. parvum, 98.8, 100, 100, and 99.7%, respectively. False-negative results for G. lamblia were obtained with specimens with low parasite numbers (n = 7) or specimens containing trophozoites only (n = 3); one specimen with a false-negative result contained numerous cysts. The one specimen false negative for C. parvum was confirmed to be positive by immunofluorescence. No cross

  20. The Effect of pH on Stability and Sorption of Cryptosporidium parvum Oocysts by Nanoparticles

    EPA Science Inventory

    Cryptosporidium parvum (C. parvum) are waterborne pathogens, which are released into the environment through infected human or animal feces. Their ability to survive outside their host organisms in harsh environmental conditions presents one of the most challenging tasks in resea...

  1. Attachment, persistence and infectivity of Cryptosporidium parvum oocysts in experimentally contaminated fruits and vegetables

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryptosporidium parvum is an environmentally resistant, abundant, and ubiquitous protozoan parasite that causes severe diarrheal disease in humans and livestock. Consumer dietary preference towards fresh and organically grown produce correlates with a heightened occurrence of foodborne outbreaks of ...

  2. EVALUATING IN VITRO INFECTIVITY FOR MEASURING UV DISINFECTION OF CRYPTOSPORIDIUM PARVUM OOCYSTS IN FINISHED WATER

    EPA Science Inventory

    UV technology to inactivate Cryptosporidium parvum oocysts has become well established in the US. The challenge now is to effectively demonstrate UV reactor performance and disinfection capacity with various finished water matrices and under different operational conditions. In s...

  3. OCCURRENCE OF 'CRYPTOSPORIDIUM' OOCYSTS IN SEWAGE EFFLUENTS AND SELECTED SURFACE WATERS

    EPA Science Inventory

    An existing method for the detection of Cryptosporidium oocysts in water was modified to investigate oocyst prevalence in large volumes of water. Surface waters and sewage effluents were filtered, eluted from the filter, and concentrated using centrifugation. The resultant pellet...

  4. Prevalence of Cryptosporidium and Giardia infections on two Ohio pig farms with different management systems.

    PubMed

    Xiao, L; Herd, R P; Bowman, G L

    1994-04-01

    The prevalence of Cryptosporidium and Giardia infections in pigs was investigated by the use of a direct immunofluorescence assay on two Ohio farms with different management systems. Cryptosporidium and Giardia infections were detected only in weanlings on the farm with slotted and wire floors, but in both weanlings and nursing piglets on the farm with porous concrete floors. Giardia infection was also detected in sows on the latter farm. The farm with porous concrete floors had a significantly higher Cryptosporidium infection rate in nursing piglets and Giardia infection rates in weanlings than the farm with slotted and wire floors. Sows were implicated as the source of both Cryptosporidium and Giardia infections for nursing piglets. PMID:8073616

  5. Prevalence and molecular characterization of Cryptosporidium spp. in dairy cattle from farms in China

    PubMed Central

    Chen, Fu

    2012-01-01

    Fecal samples of 2,056 dairy cattle from 14 farms were collected in three geographical regions of China and stained using a modified acid-fast staining technique to identify Cryptosporidium oocysts. A total of 387 (18.82%) positive samples were identified and further analyzed by polymerase chain reaction (PCR) using primers designed to amplify DNA fragments from the small subunit ribosomal RNA. The PCR products were sequenced and the sequences were deposited in the GenBank database under accession numbers EU369377-84 and GU070730-33. Phylogenetic analysis was performed and a distances matrix generated from these sequences confirmed the existence of Cryptosporidium (C.) parvum 'mouse' genotype, C. bovis, C. andersoni, C. hominis, and C. serpentis in cattle. These results represent the first report on the prevalence and genetic identification of Cryptosporidium species, and may contribute to a better understanding of the epidemiology of Cryptosporidium in cattle in China. PMID:22437531

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

    EPA Science Inventory

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

    EPA Science Inventory

    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...

  8. SPECIES AND STRAIN-SPECIFIC TYPING OF CRYPTOSPORIDIUM PARASITES IN CLINICAL AND ENVIRONMENTAL SAMPLES

    EPA Science Inventory

    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 ...

  9. EVALUATION OF CRYPTOSPORIDIUM OOCYSTS AND GIARDIA CYSTS IN A WATERSHED RESERVOIR

    EPA Science Inventory

    This investigation evaluated the occurrence of Cryptosporidium oocysts and Giardia cysts at 17 sampling locations in Lake Texoma reservoir using method 1623 with standard Envirocheck" capsule filters. The watershed serves rural agricultural communities active in cattle ranching,...

  10. EVALUATION OF CRYPTOSPORIDIUM OOCYSTS AND GIARDIA CYSTS IN A WATERSHED RESERVOIR

    EPA Science Inventory

    This investigation evaluated the occurrence of Cryptosporidium oocysts and Giardia cysts at 17 sampling locations in Lake Texoma reservoir using method 1623 with standard Envirocheck™ capsule filters. The watershed serves rural agricultural communities active in cattle ranching, ...

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

    EPA Science Inventory

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

  12. Prevalence of Cryptosporidium species in wildlife populations within a watershed landscape in southeastern New York State.

    PubMed

    Ziegler, Peter E; Wade, Susan E; Schaaf, Stephanie L; Stern, David A; Nadareski, Christopher A; Mohammed, Hussni O

    2007-06-20

    A cross-sectional study was conducted to determine the prevalence of Cryptosporidium in wildlife in the New York City (NYC) Watershed in southeastern New York State. A total of 6227 fecal samples were collected and evaluated from 5892 mammals (38 species), 263 birds (14 species), 2 reptiles (2 species), 8 amphibians (4 species), and 62 fish (15 species). Cryptosporidium was detected in 30 species. Of the species found positive for Cryptosporidium, 16 represented new records for this parasite-Alosa pseudoharengus, Larus delawarensis, Blarina brevicauda, Sorex cinereus, Parascalops breweri, Myotis lucifugus, Peromyscus maniculatus, Microtus pennsylvanicus, Clethrionomys gapperi, Tamiasciurus hudsonicus, Marmota monax, Erethizon dorsatum, Canis latrans, Mustela erminea, Mustela vison, and Lynx rufus. Factors such as age, sex, season, and land use were evaluated to determine if there was any association with infection by this parasite. Animals were more likely to be positive for Cryptosporidium during spring and in agricultural land use. PMID:17466459

  13. Molecular characterisation of species and genotypes of Cryptosporidium and Giardia and assessment of zoonotic transmission

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The molecular characterization of species and genotypes of Cryptosporidium and Giardia is essential for accurately identifying organisms and assessing zoonotic transmission. Results of recent molecular epidemiologic studies strongly suggest that zoonotic transmission plays an important role in crypt...

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

    EPA Science Inventory

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

  15. Prevalence and molecular identification of Cryptosporidium isolates from pet lizards and snakes in Italy.

    PubMed

    Rinaldi, L; Capasso, M; Mihalca, A D; Cirillo, R; Cringoli, G; Cacciò, S

    2012-11-01

    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). PMID:23193530

  16. Prevalence and molecular identification of Cryptosporidium isolates from pet lizards and snakes in Italy

    PubMed Central

    Rinaldi, L.; Capasso, M.; Mihalca, A.D.; Cirillo, R.; Cringoli, G.; Cacciò, S.

    2012-01-01

    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). PMID:23193530

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

    EPA Science Inventory

    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...

  18. CRYPTOSPORIDIUM OOCYST RECOVERY IN WATER BY EPA METHOD 1623: EVALUATION OF A MODIFIED IMS DISSOCIATION

    EPA Science Inventory

    EPA Methods 1622 and 1623 are the benchmarks for detection of Cryptosporidium spp. oocysts in water. These methods consist of filtration, elution, purification by immunomagnetic separation (IMS), and microscopic analysis after staining with a fluorescein isothiocyanate conjugate...

  19. Sensitive quantitative detection/identification of infectious Cryptosporidium parvum oocysts by signature lipid biomarker analysis

    SciTech Connect

    White, D.C. |; Alugupalli, S.; Schrum, D.P.

    1997-08-01

    Unique signature lipid biomarkers were found in the acid-fast oocytes of Cryptosporidium parvum. This makes possible the rapid detection/identification and potential infectivity directly from drinking water membrane filtrates.

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

    PubMed

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

    2011-12-01

    Infections by Cryptosporidium spp. in aquatic mammals is a major concern due to the possibility of the waterborne transmission of oocysts. The aim of the present study was to report the occurrence of Cryptosporidium spp. in Antillean manatees (Trichechus manatus) and Amazonian manatees (Trichechus inunguis) from Brazil. Fecal samples were collected and processed using Kinyoun's method. Positive samples were also submitted to the direct immunofluorescence test. The results revealed the presence of Cryptosporidium spp. oocysts in 12.5% (17/136) of the material obtained from the Antillean manatees and in 4.3% (05/115) of the samples from the Amazonian manatees. Cryptosporidium spp. infection was more prevalent in captive animals than in free-ranging specimens. PMID:22204053

  1. EFFECTS OF OZONE, CHLORINE DIOXIDE, CHLORINE, AND MONOCHLORAMINE ON CRYPTOSPORIDIUM PARVUM OOCYST VIABILITY

    EPA Science Inventory

    Purified Cryptosporidium parvum oocysts were exposed to ozone, chlorine dioxide, chlorine, and monochloramine. xcystation and mouse infectivity were comparatively evaluated to assess oocyst viability. zone and chlorine dioxide more effectively inactivated oocysts than chlorine an...

  2. Molecular epidemiological analyses of Cryptosporidium parvum virus 1 (CSpV1), a symbiotic virus of Cryptosporidium parvum, in Japan.

    PubMed

    Murakoshi, Fumi; Ichikawa-Seki, Madoka; Aita, Junya; Yaita, Seiko; Kinami, Aiko; Fujimoto, Katsuhisa; Nishikawa, Yoshifumi; Murakami, Shin; Horimoto, Taisuke; Kato, Kentaro

    2016-01-01

    We show that Cryptosporidium parvum virus 1 (CSpV1), a member of the family Partitiviridae, genus Cryspovirus that can infect Cryptosporidium parvum, is a new candidate for high-resolution tool for tracing C. parvum. CSpV1 was detected in all C. parvum-positive samples tested. Phylogenetic analysis of dsRNA1 sequence from CSpV1 can distinguish infected areas of C. parvum on the national level. Sequences detected in samples from Iwate prefecture and other islands (Tanegashima, and Okinawa) belonged to a single clade. This system can differentiate the samples from Hokkaido and south part of Japan as well as from other countries. Samples from Iwate, Tanegashima, and Okinawa belonged to a single subclade, respectively. Therefore, the CSpV1 dsRNA sequences reflect the regional distribution of their host and have potential as a high-resolution tool to trace C. parvum IIaA15G2R1 subtype. PMID:26439535

  3. Reduction of dioxin-like toxicity in effluents by additional wastewater treatment and related effects in fish.

    PubMed

    Maier, Diana; Benisek, Martin; Blaha, Ludek; Dondero, Francesco; Giesy, John P; Köhler, Heinz-R; Richter, Doreen; Scheurer, Marco; Triebskorn, Rita

    2016-10-01

    Efficiency of advanced wastewater treatment technologies to reduce micropollutants which mediate dioxin-like toxicity was investigated. Technologies compared included ozonation, powdered activated carbon and granular activated carbon. In addition to chemical analyses in samples of effluents, surface waters, sediments, and fish, (1) dioxin-like potentials were measured in paired samples of effluents, surface waters, and sediments by use of an in vitro biotest (reporter gene assay) and (2) dioxin-like effects were investigated in exposed fish by use of in vivo activity of the mixed-function, monooxygenase enzyme, ethoxyresorufin O-deethylase (EROD) in liver. All advanced technologies studied, based on degradation or adsorption, significantly reduced dioxin-like potentials in samples and resulted in lesser EROD activity in livers of fish. Results of in vitro and in vivo biological responses were not clearly related to quantification of targeted analytes by use of instrumental analyses. PMID:27262214

  4. Addition of Al and Fe salts during treatment of paper mill effluents to improve activated sludge settlement characteristics.

    PubMed

    Agridiotis, V; Forster, C F; Carliell-Marquet, C

    2007-11-01

    Metal salts, ferrous sulphate and aluminium chloride, were added to laboratory-scale activated sludge plant treating paper mill effluents to investigate the effect on settlement characteristics. Before treatment the sludge was filamentous, had stirred sludge volume index (SSVI) values in excess of 300 and was moderately hydrophobic. The use of FeSO4.7H2O took three weeks to reduce the SSVI to 90. Microscopic examination showed that Fe had converted the filamentous flocs into a compact structure. When the iron dosing was stopped, the sludge returned to its bulking state within four weeks. In a subsequent trial, the addition of AlCl3 initially resulted in an improvement of the settlement index but then caused deterioration of the sludge properties. It is possible that aluminium was overdosed and caused charge reversal, increasing the SSVI. PMID:17113285

  5. Molecular seasonal, age and gender distributions of Cryptosporidium in diarrhoeic Egyptians: distinct endemicity.

    PubMed

    El-Badry, A A; Al-Antably, A S A; Hassan, M A; Hanafy, N A; Abu-Sarea, E Y

    2015-12-01

    Cryptosporidiosis is a worldwide gastrointestinal disease caused by the protozoan Cryptosporidium parasite. It has a broad range of seasonal and age-related prevalence. We aimed to study the molecular prevalence and seasonality of Cryptosporidium over a period of 1 year in a cohort of Egyptian diarrhoeic patients. Stool samples were collected from 865 diarrhoeic patients attending outpatient clinics of Cairo University hospitals, from all age groups over a 12-month period, examined microscopically for faecal Cryptosporidium oocysts by the acid-fast staining method and for copro-DNA detection using nested polymerase chain reaction (nPCR) assays. PCR-positive samples were characterised molecularly by nPCR-restriction fragment length polymorphism (RFLP) to determine Cryptosporidium genotypes. Cryptosporidium copro-DNA was detected in 19.5% of the collected samples throughout the year, with a major peak in summer (August) and a small rise in spring (April). Infection was mainly C. hominis (95.8%) followed by C. parvum (3.0%), affecting all age groups, with predominance in the pre-school age group, and decrease with age. There were statistically significant associations between the detection of Cryptosporidium and season, diarrhoea, patient age and drinking water, while gender, contact with animals and presence of mucus in stool showed no association. Cryptosporidium in diarrhoeic Egyptians was of distinct endemicity, with the bi-model mostly influenced by population dynamics, with a clear high prevalence in pre-school children and predominating anthroponotic (C. hominis) transmission throughout the year. The obtained results highlight Cryptosporidium as a water contaminant and an important cause of health problems in Egypt, necessitating further studies of the risk factors. PMID:26440040

  6. The therapeutic effect of 16 antimicrobial agents on Cryptosporidium infection in mice.

    PubMed

    Tzipori, S R; Campbell, I; Angus, K W

    1982-04-01

    The therapeutic efficiency of 16 anti-microbial agents (Ethopabate, Nicarbazin, Sulphaquinoxaline, Furaltadone, Enterolyte-N, Sulphamethazine, Trinamide, Amprol, Phenamidine, Zoaquin, Halofuginone, Salinomycin, Monensin, Emtryl, Arprinocid and Amprolium) were examined against Cryptosporidium infections in mice. The Cryptosporidium was originally isolated from a field outbreak of calf diarrhoea. The drugs neither prevented nor modified the course of the infection as compared with infected, untreated mice. PMID:7115220

  7. Prevalence of Cryptosporidium spp. and Giardia intestinalis in swimming pools, Atlanta, Georgia.

    PubMed

    Shields, Joan M; Gleim, Elizabeth R; Beach, Michael J

    2008-06-01

    Cryptosporidium spp. and Giardia intestinalis have been found in swimming pool filter backwash during outbreaks. To determine baseline prevalence, we sampled pools not associated with outbreaks and found that of 160 sampled pools, 13 (8.1%) were positive for 1 or both parasites; 10 (6.2%) for Giardia sp., 2 (1.2%) for Cryptosporidium spp., and 1 (0.6%) for both. PMID:18507911

  8. Prevalence of Cryptosporidium spp. and Giardia intestinalis in Swimming Pools, Atlanta, Georgia

    PubMed Central

    Gleim, Elizabeth R.; Beach, Michael J.

    2008-01-01

    Cryptosporidium spp. and Giardia intestinalis have been found in swimming pool filter backwash during outbreaks. To determine baseline prevalence, we sampled pools not associated with outbreaks and found that of 160 sampled pools, 13 (8.1%) were positive for 1 or both parasites; 10 (6.2%) for Giardia sp., 2 (1.2%) for Cryptosporidium spp., and 1 (0.6%) for both. PMID:18507911

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

    PubMed

    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. PMID:27044415

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

    PubMed

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

    2011-04-01

    Groundwater from karst aquifers is an important source of drinking water worldwide. Outbreaks of cryptosporidiosis linked to surface water and treated public water are regularly reported. Cryptosporidium oocysts are resistant to conventional drinking water disinfectants and are a major concern for the water industry. Here, we examined conditions associated with oocyst transport along a karstic hydrosystem, and the impact of intensive exploitation on Cryptosporidium oocyst contamination of the water supply. We studied a well-characterized karstic hydrosystem composed of a sinkhole, a spring and a wellbore. Thirty-six surface water and groundwater samples were analyzed for suspended particulate matter, turbidity, electrical conductivity, and Cryptosporidium and Giardia (oo)cyst concentrations. (Oo)cysts were identified and counted by means of solid-phase cytometry (ChemScan RDI(®)), a highly sensitive method. Cryptosporidium oocysts were detected in 78% of both surface water and groundwater samples, while Giardia cysts were found in respectively 22% and 8% of surface water and groundwater samples. Mean Cryptosporidium oocyst concentrations were 29, 13 and 4/100 L at the sinkhole, spring and wellbore, respectively. Cryptosporidium oocysts were transported from the sinkhole to the spring and the wellbore, with respective release rates of 45% and 14%, suggesting that oocysts are subject to storage and remobilization in karst conduits. Principal components analysis showed that Cryptosporidium oocyst concentrations depended on variations in hydrological forcing factors. All water samples collected during intensive exploitation contained oocysts. Control of Cryptosporidium oocyst contamination during intensive exploitation is therefore necessary to ensure drinking water quality. PMID:21477840

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

    PubMed

    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. PMID:26921041

  12. Dissolved organic matter removal during coal slag additive soil aquifer treatment for secondary effluent recharging: Contribution of aerobic biodegradation.

    PubMed

    Wei, Liangliang; Li, Siliang; Noguera, Daniel R; Qin, Kena; Jiang, Junqiu; Zhao, Qingliang; Kong, Xiangjuan; Cui, Fuyi

    2015-06-01

    Recycling wastewater treatment plant (WWTP) effluent at low cost via the soil aquifer treatment (SAT), which has been considered as a renewable approach in regenerating potable and non-potable water, is welcome in arid and semi-arid regions throughout the world. In this study, the effect of a coal slag additive on the bulk removal of the dissolved organic matter (DOM) in WWTP effluent during SAT operation was explored via the matrix configurations of both coal slag layer and natural soil layer. Azide inhibition and XAD-resins fractionation experiments indicated that the appropriate configuration designing of an upper soil layer (25 cm) and a mixture of soil/coal slag underneath would enhance the removal efficiency of adsorption and anaerobic biodegradation to the same level as that of aerobic biodegradation (31.7% vs 32.2%), while it was only 29.4% compared with the aerobic biodegradation during traditional 50 cm soil column operation. The added coal slag would preferentially adsorb the hydrophobic DOM, and those adsorbed organics could be partially biodegraded by the biomass within the SAT systems. Compared with the relatively lower dissolved organic carbon (DOC), ultraviolet light adsorption at 254 nm (UV-254) and trihalomethane formation potential (THMFP) removal rate of the original soil column (42.0%, 32.9%, and 28.0%, respectively), SSL2 and SSL4 columns would enhance the bulk removal efficiency to more than 60%. Moreover, a coal slag additive in the SAT columns could decline the aromatic components (fulvic-like organics and tryptophan-like proteins) significantly. PMID:25845997

  13. Additive Manufacturing of 17-4 PH Stainless Steel: Post-processing Heat Treatment to Achieve Uniform Reproducible Microstructure

    NASA Astrophysics Data System (ADS)

    Cheruvathur, Sudha; Lass, Eric A.; Campbell, Carelyn E.

    2016-03-01

    17-4 precipitation hardenable (PH) stainless steel is a useful material when a combination of high strength and good corrosion resistance up to about 315°C is required. In the wrought form, this steel has a fully martensitic structure that can be strengthened by precipitation of fine Cu-rich face-centered cubic phase upon aging. When fabricated via additive manufacturing (AM), specifically laser powder-bed fusion, 17-4 PH steel exhibits a dendritic structure containing a substantial fraction of nearly 50% of retained austenite along with body centered cubic/martensite and fine niobium carbides preferentially aligned along interdendritic boundaries. The effect of post-build thermal processing on the material microstructure is studied in comparison to that of conventionally produced wrought 17-4 PH with the intention of creating a more uniform, fully martensitic microstructure. The recommended stress relief heat treatment currently employed in industry for post-processing of AM 17-4 PH steel is found to have little effect on the as-built dendritic microstructure. It is found that, by implementing the recommended homogenization heat treatment regimen of Aerospace Materials Specification 5355 for CB7Cu-1, a casting alloy analog to 17-4 PH, the dendritic solidification structure is eliminated, resulting in a microstructure containing about 90% martensite with 10% retained austenite.

  14. Molecular characterization of Cryptosporidium in bats from Yunnan province, southwestern China.

    PubMed

    Wang, Wei; Cao, Lili; He, Biao; Li, Jiping; Hu, Tingsong; Zhang, Fuqiang; Fan, Quanshui; Tu, Changchun; Liu, Quan

    2013-12-01

    The present study was conducted to investigate the prevalence and genotypes of Cryptosporidium in bats. A total of 247 bats, belonging to Rhinolophus sinicus , Rousettus leschenaultia, Aselliscus stoliczkanus , and Hipposideros fulvus , were collected in Yunnan Province, Southwestern China, and the intestinal tissues were examined for Cryptosporidium infection by PCR amplification of the small subunit ribosomal RNA (SSU rRNA). The overall infection rate was 7.7% (95% CI, 4.5 to 11.0%), with R. sinicus having the highest level at 9.5% (95% CI, 2.8 to 16.1%) followed by A. stoliczkanus at 7.8% (95% CI, 2.2 to 18.9%), H. fulvus at 7.2% (95% CI, 1.1 to 13.4%), and R. leschenaultia at 5.7% (95% CI, 1.2 to 15.7%). DNA sequence and phylogenetic analyses based on SSU rRNA revealed the presence of 2 novel genotypes, designated as Cryptosporidium bat genotype I in A. stoliczkanus and R. sinicus and Cryptosporidium bat genotype II in R. leschenaultia, R. sinicus , and H. fulvus . This is the first report of Cryptosporidium genotypes in bats. Further biological and genetic characterization is needed to determine the relationship of the 2 novel genotypes to established Cryptosporidium species-genotypes. PMID:23886252

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

    PubMed

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

    2015-01-01

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

  16. Prevalence of the Cryptosporidium Pig Genotype II in Pigs from the Yangtze River Delta, China

    PubMed Central

    Yuan, Zhongying; Lu, Weiyuan; Xu, Yuxin; Cao, Jianping

    2011-01-01

    Background Cryptosporidium spp. is prevalent globally, pigs are an important Cryptosporidium reservoir. In China, little data regarding rates of Cryptosporidium infections in pigs are available. The present study was therefore aimed at characterizing the distribution of Cryptosporidium species in pigs from two different cities, Shaoxing and Shanghai, from the Yangtze River delta. Methodology/Principal Findings Nested PCR to amplify the 18S rRNA locus on DNA extracted from fecal samples (n = 94) revealed the positive rate of Cryptosporidium in pigs from two cities was approximately 17.0%. The positive rates in Shanghai and Shaoxing were 14.3% and 25.0% respectively. Amplified sequences were verified by sequencing. The identified strain belonged to the C. pig genotype II using BLAST analysis in the NCBI database. Conclusion/Significance Our finding of Cryptosporidium pig genotype II in pigs in the Yangtze River delta area suggests that pig farms in this region must be considered a public health threat and proper control measures be introduced. PMID:21677776

  17. Genotypic Characterization of Cryptosporidium hominis from Water Samples in São Paulo, Brazil

    PubMed Central

    Araújo, Ronalda S.; Dropa, Milena; Fernandes, Licia N.; Carvalho, Terezinha T.; Sato, Maria Inês Z.; Soares, Rodrigo M.; Matté, Glavur R.; Matté, Maria Helena

    2011-01-01

    The protozoan parasite Cryptosporidium has emerged as one of the most important water contaminants, causing waterborne outbreaks of diarrheal diseases worldwide. The small size of oocysts under the microscope and the possibility of changes in characteristics of oocysts, mainly in environmental samples, make the taxonomy of the genus difficult if morphologic characteristics are considered. This limitation encouraged the application of molecular methods to identify this microorganism. The aim of this study was to detect and identify by nested-polymerase chain reaction oocysts of Cryptosporidium present in water samples in the state of São Paulo, Brazil. Water samples were concentrated through a membrane filter, DNA was extracted by using a standard technique, and both amplification reactions used forward and reverse oligonucleotides that were complementary to Cryptosporidium 18S ribosomal RNA gene sequences. Thirty water samples from different sites of collection in the state of São Paulo were evaluated. Cryptosporidium oocysts were detected in 30% of the samples. By genoptyping, C. hominis and Cryptosporidium sp. were identified in recreational water and C. meleagridis was identified in surface water samples. This is the first report of C. hominis in environmental samples in Brazil. Although identification of Cryptosporidium is still a difficult task, molecular methods are essential for specific identification and are a helpful tool to aid to understand the epidemiology of this parasite in Brazil. PMID:22049036

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

    PubMed Central

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

    2015-01-01

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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

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

    2016-01-01

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

  1. Addition of hydrogen peroxide for the simultaneous control of bromate and odor during advanced drinking water treatment using ozone.

    PubMed

    Wang, Yongjing; Yu, Jianwei; Zhang, Dong; Yang, Min

    2014-03-01

    Complete removal of the characteristic septic/swampy odor from Huangpu River source water could only be achieved under an ozone dose as high as 4.0 mg/L in an ozone-biological activated carbon (O3-BAC) process, which would lead to the production of high concentrations of carcinogenic bromate due to the high bromide content. This study investigated the possibility of simultaneous control of bromate and the septic/swampy odor by adding H2O2 prior to the O3-BAC process for the treatment of Huangpu River water. H2O2 addition could reduce the bromate concentration effectively at an H2O2/O3 (g/g) ratio of 0.5 or higher. At the same time, the septic/swampy odor removal was enhanced by the addition of H2O2, although optimization of the H2O2/O3 ratio was required for each ozone dose. At an ozone dose of 2.0 mg/L, the odor was removed completely at an H2O2/O3 ratio of 0.5. The results indicated that H2O2 application at a suitable dose could enhance the removal of the septic/swampy odor while suppressing the formation of bromate during ozonation of Huangpu River source water. PMID:25079267

  2. Effect of supplementary carbon addition in the treatment of low C/N high-technology industrial wastewater by MBR.

    PubMed

    Kumar, Mathava; Lee, Pei-Yun; Fukusihma, Toshikazu; Whang, Liang-Ming; Lin, Jih-Gaw

    2012-06-01

    The effect of supplementary carbon addition for the treatment of high-technology industrial wastewater in a membrane bioreactor (MBR) was investigated. The MBR was operated for 302 days under different C/N (BOD(L)/NH(4)(+)-N) ratios, i.e. 0.9-1 to 20 days, 1.6-21 to 42 days, 2.9-43 to 82 days, 3.6-83 to 141 days, 4.8-165 to 233 days and 9.3-240 to 302 days. Irrespective of the C/N ratios investigated, SS and BOD(5) removal efficiencies were above 95% and above 80% COD removal efficiency was observed. In addition, complete nitrification was observed throughout the investigation. However, denitrification and total nitrogen removal efficiencies reached their maximum values at the highest C/N ratio (9.3) investigated. Real-time PCR analysis revealed 10 times higher ammonia oxidizing bacteria to total bacteria ratio under the highest C/N ratio condition (9.3) compared to the low C/N ratio condition (1.6). PMID:22300636

  3. An Irish perspective on Cryptosporidium. Part 1

    PubMed Central

    2006-01-01

    Cryptosporidiosis, a protozoal disease which causes significant morbidity in humans, is one of the chief causes of diarrhoea in neonatal ruminants. Although the parasite poses a significant threat to public health and animal health in Ireland, its epidemiology on the island is only poorly understood. Environmental studies have shown the waterborne parasite to be widespread in some untreated waterbodies around Ireland. The island's hydrogeological situation, combined with high stocking rates of livestock and the absence of filtration from regular water treatment, render it vulnerable to large-scale outbreaks. This review discusses the parasite in the Irish context and underlines the need for a reference facility to provide active surveillance on the island. PMID:21851685

  4. Prevalence and Genetic Diversity of Giardia duodenalis and Cryptosporidium spp. among School Children in a Rural Area of the Amhara Region, North-West Ethiopia

    PubMed Central

    de Lucio, Aida; Amor-Aramendía, Aranzazu; Bailo, Begoña; Saugar, José M.; Anegagrie, Melaku; Arroyo, Ana; López-Quintana, Beatriz; Zewdie, Derjew; Ayehubizu, Zimmam; Yizengaw, Endalew; Abera, Bayeh; Yimer, Mulat; Mulu, Wondemagen; Hailu, Tadesse; Herrador, Zaida; Fuentes, Isabel; Carmena, David

    2016-01-01

    Backgroud Giardia duodenalis and Cryptosporidium spp. are enteric protozoan causing gastrointestinal illness in humans and animals. Giardiasis and cryptosporidiosis are not formally considered as neglected tropical diseases, but belong to the group of poverty-related infectious diseases that impair the development and socio-economic potential of infected individuals in developing countries. Methods We report here the prevalence and genetic diversity of G. duodenalis and Cryptosporidium spp. in children attending rural primary schools in the Bahir Dar district of the Amhara Region, Ethiopia. Stool samples were collected from 393 children and analysed by molecular methods. G. duodenalis was detected by real-time PCR, and the assemblages and sub-assemblages were determined by multilocus sequence-based genotyping of the glutamate dehydrogenase and β-giardin genes of the parasite. Detection and identification of Cryptosporidium species was carried out by sequencing of a partial fragment of the small-subunit ribosomal RNA gene. Principal Findings The PCR-based prevalences of G. duodenalis and Cryptosporidium spp. were 55.0% (216/393) and 4.6% (18/393), respectively. A total of 78 G. duodenalis isolates were successfully characterized, revealing the presence of sub-assemblages AII (10.3%), BIII (28.2%), and BIV (32.0%). Discordant typing results AII/AIII and BIII/BIV were identified in 7.7% and 15.4% of the isolates, respectively. An additional five (6.4%) isolates were assigned to assemblage B. No mixed infections of assemblages A+B were found. Extensive genetic variation at the nucleotide level was observed within assemblage B (but no within assemblage A), resulting in the identification of a large number of sub-types. Cryptosporidium diversity was demonstrated by the occurrence of C. hominis, C. parvum, and C. viatorum in the population under study. Conclusions Our data suggest an epidemiological scenario with an elevated transmission intensity of a wide range of G

  5. EPA ORD/EPA REGION 2/PITTSBURGH WSA COOPERATIVE EFFORT: EVALUATING CRYPTOSPORIDIUM AND GIARDIA IN COMBINED SEWER OVERFLOW AS A THREAT TO DRINKING WATER SUPPLIES

    EPA Science Inventory

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

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

    EPA Science Inventory

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

  7. Flow cytometric detection of Cryptosporidium oocysts in human stool samples.

    PubMed Central

    Valdez, L M; Dang, H; Okhuysen, P C; Chappell, C L

    1997-01-01

    Cryptosporidium parvum is an important pathogen that causes diarrhea in virtually all human populations. Improved diagnostic methods are needed to understand the risk factors, modes of transmission, and impact of cryptosporidiosis. In the present study, we fluorescently labeled and counted C. parvum oocysts by flow cytometry (FC) and developed a simple and efficient method of processing human stool samples for FC analysis. Formed stool (suspended in phosphate-buffered saline) from an asymptomatic, healthy individual was seeded with known concentrations of oocysts, and oocysts were labeled with a cell wall-specific monoclonal antibody and detected by FC. The method described herein resulted in a mean oocyst recovery rate of 45% +/- 16% (median, 42%), which consistently yielded a fourfold increase in sensitivity compared to direct fluorescent-antibody assay of seeded stool samples. However, in many instances, FC detected as few as 10(3) oocysts per ml. Thus, FC provides a reproducible and sensitive method for C. parvum oocyst detection. PMID:9230372

  8. Early Treatment with Addition of Low Dose Prednisolone to Methotrexate Improves Therapeutic Outcome in Severe Psoriatic Arthritis

    PubMed Central

    Mahajan, Vikram K; Sharma, Anju Lath; Chauhan, Pushpinder S; Mehta, Karaninder S; Sharma, Nand Lal

    2013-01-01

    Psoriatic arthritis (PsA) is increasingly being recognized to cause progressive joint damage and disability. PsA unresponsive to non-steroidal anti-inflammatory drugs (NSAIDs), the conventional first-line choice of treatment, is usually managed with disease-modifying antirheumatic drugs (DMARDs) especially methotrexate. An 18-year-old HIV-negative male had progressively severe PsA of 4-month duration that was nearly confining him to a wheel chair. He did not respond to multiple NSAIDs, alone or in combination with methotrexate (15 mg/week), given for 4 weeks. Addition of prednisolone (10 mg on alternate days) controlled his symptoms within a week. The NSAIDs could be withdrawn after 4 weeks as the treatment progressed. The doses were tapered for methotrexate (5 mg/week) and prednisolone (2.5 mg on alternate days) every 8 weekly subsequently during 15 months of follow-up without recurrence/deformities or drug toxicity. For years, the use of corticosteroids in psoriasis has been criticized for their propensity to exacerbate the skin disease on withdrawal. However, monitored use of corticosteroids, even in low doses, combined with DMARDs may be a good therapeutic option in early stage of the PsA rather than ‘steroid rescue’ later. This will help in early control of joint inflammation, prevent joint damage and maintain long-term good functional capacity and quality of life. This may be useful when the cost or availability of biologics precludes their use. However, we discourage the use of corticosteroids as monotherapy. PMID:23723489

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

    PubMed

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

    2013-03-31

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

  10. Subtyping Novel Zoonotic Pathogen Cryptosporidium Chipmunk Genotype I

    PubMed Central

    Guo, Yaqiong; Cebelinski, Elizabeth; Matusevich, Christine; Alderisio, Kerri A.; Lebbad, Marianne; McEvoy, John; Roellig, Dawn M.; Yang, Chunfu

    2015-01-01

    Cryptosporidium chipmunk genotype I is an emerging zoonotic pathogen in humans. The lack of subtyping tools makes it impossible to determine the role of zoonotic transmission in epidemiology. To identify potential subtyping markers, we sequenced the genome of a human chipmunk genotype I isolate. Altogether, 9,509,783 bp of assembled sequences in 853 contigs were obtained, with an N50 of 117,886 bp and >200-fold coverage. Based on the whole-genome sequence data, two genetic markers encoding the 60-kDa glycoprotein (gp60) and a mucin protein (ortholog of cgd1_470) were selected for the development of a subtyping tool. The tool was used for characterizing chipmunk genotype I in 25 human specimens from four U.S. states and Sweden, one specimen each from an eastern gray squirrel, a chipmunk, and a deer mouse, and 4 water samples from New York. At the gp60 locus, although different subtypes were seen among the animals, water, and humans, the 15 subtypes identified differed mostly in the numbers of trinucleotide repeats (TCA, TCG, or TCT) in the serine repeat region, with only two single nucleotide polymorphisms in the nonrepeat region. Some geographic differences were found in the subtype distribution of chipmunk genotype I from humans. In contrast, only two subtypes were found at the mucin locus, which differed from each other in the numbers of a 30-bp minisatellite repeat. Thus, Cryptosporidium chipmunk genotype I isolates from humans and wildlife are genetically similar, and zoonotic transmission might play a potential role in human infections. PMID:25762767

  11. Insulin receptor isoform A and insulin-like growth factor II as additional treatment targets in human osteosarcoma.

    PubMed

    Avnet, Sofia; Sciacca, Laura; Salerno, Manuela; Gancitano, Giovanni; Cassarino, Maria Francesca; Longhi, Alessandra; Zakikhani, Mahvash; Carboni, Joan M; Gottardis, Marco; Giunti, Armando; Pollak, Michael; Vigneri, Riccardo; Baldini, Nicola

    2009-03-15

    Despite the frequent presence of an insulin-like growth factor I receptor (IGFIR)-mediated autocrine loop in osteosarcoma (OS), interfering with this target was only moderately effective in preclinical studies. Here, we considered other members of the IGF system that might be involved in the molecular pathology of OS. We found that, among 45 patients with OS, IGF-I and IGFBP-3 serum levels were significantly lower, and IGF-II serum levels significantly higher, than healthy controls. Increased IGF-II values were associated with a decreased disease-free survival. After tumor removal, both IGF-I and IGF-II levels returned to normal values. In 23 of 45 patients, we obtained tissue specimens and found that all expressed high mRNA level of IGF-II and >IGF-I. Also, isoform A of the insulin receptor (IR-A) was expressed at high level in addition to IGFIR and IR-A/IGFIR hybrids receptors (HR(A)). These receptors were also expressed in OS cell lines, and simultaneous impairment of IGFIR, IR, and Hybrid-Rs by monoclonal antibodies, siRNA, or the tyrosine kinase inhibitor BMS-536924, which blocks both IGFIR and IR, was more effective than selective anti-IGFIR strategies. Also, anti-IGF-II-siRNA treatment in low-serum conditions significantly inhibited MG-63 OS cells that have an autocrine circuit for IGF-II. In summary, IGF-II rather than IGF-I is the predominant growth factor produced by OS cells, and three different receptors (IR-A, HR(A), and IGFIR) act complementarily for an IGF-II-mediated constitutive autocrine loop, in addition to the previously shown IGFIR/IGF-I circuit. Cotargeting IGFIR and IR-A is more effective than targeting IGF-IR alone in inhibiting OS growth. PMID:19258511

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

    PubMed Central

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

    2015-01-01

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

  13. Investigation on the eco-toxicity of lake sediments with the addition of drinking water treatment residuals.

    PubMed

    Yuan, Nannan; Wang, Changhui; Pei, Yuansheng

    2016-08-01

    Drinking water treatment residuals (WTRs) have a potential to realize eutrophication control objectives by reducing the internal phosphorus (P) load of lake sediments. Information regarding the ecological risk of dewatered WTR reuse in aquatic environments is generally lacking, however. In this study, we analyzed the eco-toxicity of leachates from sediments with or without dewatered WTRs toward algae Chlorella vulgaris via algal growth inhibition testing with algal cell density, chlorophyll content, malondialdehyde content, antioxidant enzyme superoxide dismutase activity, and subcellular structure indices. The results suggested that leachates from sediments unanimously inhibited algal growth, with or without the addition of different WTR doses (10% or 50% of the sediment in dry weight) at different pH values (8-9), as well as from sediments treated for different durations (10 or 180days). The inhibition was primarily the result of P deficiency in the leachates owing to WTR P adsorption, however, our results suggest that the dewatered WTRs were considered as a favorable potential material for internal P loading control in lake restoration projects, as it shows acceptably low risk toward aquatic plants. PMID:27521931

  14. Effect of additional heat treatment of 2024-T3 on the growth of fatigue crack in air and in vacuum

    NASA Technical Reports Server (NTRS)

    Louwaard, E. P.

    1986-01-01

    In order to determine the influence of ductility on the fatigue crack growth rate of aluminum alloys, fatigue tests were carried out on central notched specimens of 2024-T3 and 2024-T8 sheet material. The 2024-T8 material was obtained by an additional heat treatment applied on 2024-T3 (18 hours at 192 C), which increased the static yield strength from 43.6 to 48.9 kgf/sq mm. A change in the ultimate strength was not observed. Fatigue tests were carried out on both materials in humid air and in high vacuum. According to a new crack propagation model, crack extension is supported to be caused by a slip-related process and debonding triggered by the environment. This model predicts an effect of the ductility on the crack growth rate which should be smaller in vacuum than in humid air; however, this was not confirmed. In humid air the crack-growth rate in 2024-T8 was about 2 times faster than in 2024-T3, while in vacuum the ratio was about 2.5. Crack closure measurements gave no indications that crack closure played a significant role in both materials. Some speculative explanations are briefly discussed.

  15. Reduction of sludge generation by the addition of support material in a cyclic activated sludge system for municipal wastewater treatment.

    PubMed

    Araujo, Moacir Messias de; Lermontov, André; Araujo, Philippe Lopes da Silva; Zaiat, Marcelo

    2013-09-01

    An innovative biomass carrier (Biobob®) was tested for municipal wastewater treatment in an activated sludge system to evaluate the pollutant removal performance and the sludge generation for different carrier volumes. The experiment was carried out in a pilot-scale cyclic activated sludge system (CASS®) built with three cylindrical tanks in a series: an anoxic selector (2.1 m(3)), an aerobic selector (2.5 m(3)) and the main aerobic reactor (25.1 m(3)). The results showed that by adding the Biobob® carrier decreased the MLVSS concentration, which consequently reduced the waste sludge production of the system. Having 7% and 18% (v/v) support material in the aerobic reactor, the observed biomass yield decreased 18% and 36%, respectively, relative to the reactor operated with suspended biomass. The addition of media did not affect the system's performance for COD and TSS removal. However, TKN and TN removal were improved by 24% and 14%, respectively, using 18% (v/v) carrier. PMID:23831747

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

    PubMed

    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. PMID:23886616

  17. Cryptosporidium suis infection in post-weaned and adult pigs in Shaanxi province, northwestern China.

    PubMed

    Lin, Qing; Wang, Xing-Ye; Chen, Jian-Wen; Ding, Ling; Zhao, Guang-Hui

    2015-02-01

    Cryptosporidium spp., ubiquitous enteric parasitic protozoa of vertebrates, recently emerged as an important cause of economic loss and zoonosis. The present study aimed to determine the distribution and species of Cryptosporidium in post-weaned and adult pigs in Shaanxi province, northwestern China. A total of 1,337 fresh fecal samples of post-weaned and adult pigs were collected by sterile disposable gloves from 8 areas of Shaanxi province. The samples were examined by Sheather's sugar flotation technique and microscopy at × 400 magnification for Cryptosporidium infection, and the species in positive samples was further identified by PCR amplification of the small subunit (SSU) rRNA gene. A total of 44 fecal samples were successfully amplified by the nested PCR of the partial SSU rRNA, with overall prevalence of 3.3%. The average prevalence of Cryptosporidium infection in each pig farms ranged from 0 to 14.4%. Species identification by sequencing of SSU rRNA gene revealed that 42 (3.1%) samples were Cryptosporidium suis and 2 (0.15%) were Cryptosporidium scrofarum. C. suis had the highest prevalence (7.5%) in growers and the lowest in breeding pigs (0.97%). C. suis was the predominant species in pre-weaned and adult pigs, while C. scrofarum infected pigs older than 3 months only. A season-related difference of C. suis was observed in this study, with the highest prevalence in autumn (5.5%) and the lowest (1.7%) in winter. The present study provided basic information for control of Cryptosporidium infection in pigs and assessment of zoonotic transmission of pigs in Shaanxi province, China. PMID:25748718

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

    PubMed

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

    2016-01-01

    The identification and characterisation of Cryptosporidium genotypes and subtypes are fundamental to the study of cryptosporidiosis epidemiology, aiding in prevention and control strategies. The objective was to determine the genetic diversity of Cryptosporidium 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 for Cryptosporidium 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

  19. Cryptosporidium suis Infection in Post-Weaned and Adult Pigs in Shaanxi Province, Northwestern China

    PubMed Central

    Lin, Qing; Wang, Xing-Ye; Chen, Jian-Wen; Ding, Ling; Zhao, Guang-Hui

    2015-01-01

    Cryptosporidium spp., ubiquitous enteric parasitic protozoa of vertebrates, recently emerged as an important cause of economic loss and zoonosis. The present study aimed to determine the distribution and species of Cryptosporidium in post-weaned and adult pigs in Shaanxi province, northwestern China. A total of 1,337 fresh fecal samples of post-weaned and adult pigs were collected by sterile disposable gloves from 8 areas of Shaanxi province. The samples were examined by Sheather’s sugar flotation technique and microscopy at×400 magnification for Cryptosporidium infection, and the species in positive samples was further identified by PCR amplification of the small subunit (SSU) rRNA gene. A total of 44 fecal samples were successfully amplified by the nested PCR of the partial SSU rRNA, with overall prevalence of 3.3%. The average prevalence of Cryptosporidium infection in each pig farms ranged from 0 to 14.4%. Species identification by sequencing of SSU rRNA gene revealed that 42 (3.1%) samples were Cryptosporidium suis and 2 (0.15%) were Cryptosporidium scrofarum. C. suis had the highest prevalence (7.5%) in growers and the lowest in breeding pigs (0.97%). C. suis was the predominant species in pre-weaned and adult pigs, while C. scrofarum infected pigs older than 3 months only. A season-related difference of C. suis was observed in this study, with the highest prevalence in autumn (5.5%) and the lowest (1.7%) in winter. The present study provided basic information for control of Cryptosporidium infection in pigs and assessment of zoonotic transmission of pigs in Shaanxi province, China. PMID:25748718

  20. Immune response of newborn BALB/c mice to Cryptosporidium infection.

    PubMed

    Ahmadian, Nasser; Pashaei-Asl, Roghiyeh; Ahmadian, Masomeh; Rahmati-Yamchi, Mohammad; Shahabi, Saed; Vazini, Hossein

    2016-09-01

    Cryptosporidium parvum is a protozoan parasite which causes diarrheal in human and animals worldwide. Infection transmission has reported through oral-fecal by infectious objects through foods and drinks. In this study we explored the immune response pathway in animal model for C. parvum to develop the new treatment way. Oocysts collected from fecal positive for C. parvum and diluted about 1:5 in sucrose solution. New born BALB/c mice (3 days) divided to 2 different groups. Control group hadn't received any oocyst, the test groups received 5 × 10(5) oocysts. 5 mice selected for each control group and 11 mice chosen for each test group. Blood collected from heart bleeds in days of 6, 9, 12 and 16. Protein concentrations determined by bio-photometer. Dot blotting used to find out total antibody concentrations oocyst antigen. Among the test and the control groups, blots appeared in test group which means antibody production, but not any blot observed in the control groups. The non-characteristic proteins in serum were measured by the biophotometer. In this study, we investigated antibody serum production against C. parvum oocysts in new born BALB/c mice. The detected antibody through dot blot technique was our aims which had conjugated to our characteristic antiserum. The recorded numbers for the controls by biophotometer related to the non characteristic proteins in serum. The results of this study can used to produce polyclonal or monoclonal antibodies against cryptosporidiosis. PMID:27605838

  1. Efficacy of using harmless Bacillus endospores to estimate the inactivation of Cryptosporidium parvum oocysts in water.

    PubMed

    Garvey, Mary; Clifford, Eoghan; O'Reilly, Edmond; Rowan, Neil J

    2013-06-01

    The need to use complex in vitro cell culture, expensive equipment, and highly-trained technicians that are available only to specialist laboratories has significantly limited studies assessing the potential of pulsed UV light (PUV) to inactivate the waterborne parasite Cryptosporidium parvum in drinking water. This constitutes the first study to report on the use of different non-pathogenic Bacillus endospores as potential surrogate organisms to indicate the PUV inactivation performance of a C. parvum oocyst suspended in water. Findings showed that PUV effectively inactivated approximately 5 log10 CFU/ml Bacillus megaterium and Bacillus pumilus endospores suspended in water at a UV dose of 9.72 μJ/cm(2) that also inactivated statistically similar levels of C. parvum oocysts (P < 0.05), as determined by combined in vitro HCT-8 cell culture and quantitative PCR. Specifically, this study demonstrated that B. megaterium exhibited greater or similar PUV-inactivation kinetic data compared to that of similarly treated C. parvum over the UV dose range 6.4 to 12.9 μJ/cm(2). Therefore, the former may be used as an indicator organism for safely investigating the PUV-inactivation performance of this chlorine-resistant, waterborne parasite at the waste-water treatment plant level. Findings presented will impact positively on future water quality studies and on public health. PMID:23145570

  2. ANTIPARASITIC ACTIVITY OF SILVER AND COPPER OXIDE NANOPARTICLES AGAINST ENTAMOEBA HISTOLYTICA AND CRYPTOSPORIDIUM PARVUM CYSTS.

    PubMed

    Saad, Halim A; Soliman, Mohamed I; Azzam, Ahmed M; Mostafa, B

    2015-12-01

    Nanoparticles (NPs) have received more attention as antiparasitic agents. In the present study, silver and copper nanoparticles were synthesized and characterized using scanning electron microscopy (SEM), transmission electron microscope (TEM) and X-ray fluorescence (XRF). The antiparasitic activity of Ag and CuO nanoparticles were tested against two of the most environmentally spread parasites in Egypt (Entamoeba histolytica and Cryptosporidium parvum). The average sizes of synthesized Ag NPs and CuO NPs were 9 & 29 nm respectively and a significant reduction for cysts viability (p > 0.05) was observed for CuO NPs against E. histolytica cysts and Ag NPs against C. parvum oocysts. Moreover, LC50-3h of CuO NPs for E. histolytica and C. parvum were 0.13 and 0.72 mg/l, while Ag NPs recorded 0.34 and 0.54 mg/l respectively. Accordingly, these NPs could be suggested as a new nanoform agent for safe and effective treatment of E. histolytica and C. parvum parasites. PMID:26939237

  3. Detection of Viable Cryptosporidium parvum in Soil by Reverse Transcription–Real-Time PCR Targeting hsp70 mRNA ▿

    PubMed Central

    Liang, Zhanbei; Keeley, Ann

    2011-01-01

    Extraction of high-quality mRNA from Cryptosporidium parvum is a key step in PCR detection of viable oocysts in environmental samples. Current methods for monitoring oocysts are limited to water samples; therefore, the goal of this study was to develop a rapid and sensitive procedure for Cryptosporidium detection in soil samples. The efficiencies of five RNA extraction methods were compared (mRNA extraction with the Dynabeads mRNA Direct kit after chemical and physical sample treatments, and total RNA extraction methods using the FastRNA Pro Soil-Direct, PowerSoil Total RNA, E.Z.N.A. soil RNA, and Norgen soil RNA purification kits) for the direct detection of Cryptosporidium with oocyst-spiked sandy, loamy, and clay soils by using TaqMan reverse transcription-PCR. The study also evaluated the presence of inhibitors by synthesis and incorporation of an internal positive control (IPC) RNA into reverse transcription amplifications, used different facilitators (bovine serum albumin, yeast RNA, salmon DNA, skim milk powder, casein, polyvinylpyrrolidone, sodium hexametaphosphate, and Salmonella enterica serovar Typhi) to mitigate RNA binding on soil components, and applied various treatments (β-mercaptoethanol and bead beating) to inactivate RNase and ensure the complete lysis of oocysts. The results of spiking studies showed that Salmonella cells most efficiently relieved binding of RNA. With the inclusion of Salmonella during extraction, the most efficient mRNA method was Dynabeads, with a detection limit of 6 × 102 oocysts g−1 of sandy soil. The most efficient total RNA method was PowerSoil, with detection limits of 1.5 × 102, 1.5 × 103, and 1.5 × 104 C. parvum oocysts g−1 soil for sandy, loamy, and clay samples, respectively. PMID:21803904

  4. A Screening Pipeline for Antiparasitic Agents Targeting Cryptosporidium Inosine Monophosphate Dehydrogenase

    PubMed Central

    Sharling, Lisa; Liu, Xiaoping; Gollapalli, Deviprasad R.; Maurya, Sushil K.; Hedstrom, Lizbeth; Striepen, Boris

    2010-01-01

    Background The protozoan parasite Cryptosporidium parvum is responsible for significant disease burden among children in developing countries. In addition Cryptosporidiosis can result in chronic and life-threatening enteritis in AIDS patients, and the currently available drugs lack efficacy in treating these severe conditions. The discovery and development of novel anti-cryptosporidial therapeutics has been hampered by the poor experimental tractability of this pathogen. While the genome sequencing effort has identified several intriguing new targets including a unique inosine monophosphate dehydrogenase (IMPDH), pursuing these targets and testing inhibitors has been frustratingly difficult. Methodology and Principal Findings Here we have developed a pipeline of tools to accelerate the in vivo screening of inhibitors of C. parvum IMPDH. We have genetically engineered the related parasite Toxoplasma gondii to serve as a model of C. parvum infection as the first screen. This assay provides crucial target validation and a large signal window that is currently not possible in assays involving C. parvum. To further develop compounds that pass this first filter, we established a fluorescence-based assay of host cell proliferation, and a C. parvum growth assay that utilizes automated high-content imaging analysis for enhanced throughput. Conclusions and Significance We have used these assays to evaluate C. parvum IMPDH inhibitors emerging from our ongoing medicinal chemistry effort and have identified a subset of 1,2,3-triazole ethers that exhibit excellent in vivo selectivity in the T. gondii model and improved anti-cryptosporidial activity. PMID:20706578

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

    USGS Publications Warehouse

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

    1997-01-01

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

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

    PubMed

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

    2014-12-15

    Protozoan pathogens present a significant human health concern, and prevention of contamination into potable networks remains a key focus for drinking water providers. Here, we monitored the change in Cryptosporidium concentration in source water during high flow events in a multi-use catchment. Furthermore, we investigated the diversity of Cryptosporidium species/genotypes present in the source water, and delivered an oocyst infectivity fraction. There was a positive and significant correlation between Cryptosporidium concentration and flow (ρ = 0.756) and turbidity (ρ = 0.631) for all rainfall-runoff events, despite variable source water pathogen concentrations. Cell culture assays measured oocyst infectivity and suggested an overall source water infectious fraction of 3.1%. No infectious Cryptosporidium parvum or Cryptosporidium hominis were detected, although molecular testing detected C. parvum in 7% of the samples analysed using PCR-based molecular techniques. Twelve Cryptosporidium species/genotypes were identified using molecular techniques, and were reflective of the host animals typically found in remnant vegetation and agricultural areas. The inclusion of molecular approaches to identify Cryptosporidium species and genotypes highlighted the diversity of pathogens in water, which originated from various sources across the catchment. We suggest this mixing of runoff water from a range of landuses containing diverse Cryptosporidium hosts is a key explanation for the often-cited difficulty forming strong pathogen-indicator relationships. PMID:25306487

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

    EPA Science Inventory

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

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

    PubMed Central

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

    1997-01-01

    For the first time, Cryptosporidium sp. oocysts were identified in fecal and intestinal samples from free-ranging marine turtles, Chelonia mydas, from the Hawaiian Islands. The oocysts produced positive reactions with commercial test kits recommended for the detection of human-infectious waterborne oocysts of Cryptosporidium parvum. PMID:16535658

  9. 40 CFR 141.211 - Special notice for repeated failure to conduct monitoring of the source water for Cryptosporidium...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 23 2011-07-01 2011-07-01 false Special notice for repeated failure to conduct monitoring of the source water for Cryptosporidium and for failure to determine bin classification or mean Cryptosporidium level. 141.211 Section 141.211 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS...

  10. 40 CFR 141.211 - Special notice for repeated failure to conduct monitoring of the source water for Cryptosporidium...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 22 2010-07-01 2010-07-01 false Special notice for repeated failure to conduct monitoring of the source water for Cryptosporidium and for failure to determine bin classification or mean Cryptosporidium level. 141.211 Section 141.211 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS...

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

    PubMed

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

    2015-12-15

    Cryptosporidium is an enteric parasite that infects humans and a wide range of animals. Relatively little is known about the epidemiology and taxonomy of Cryptosporidium in fish. In the present study, a total of 775 fish, belonging to 46 species and comprising ornamental fish, marine fish and freshwater fish were screened for the prevalence of Cryptosporidium by PCR. The overall prevalence of Cryptosporidium in fish was 5.3% (41/775), with prevalences ranging from 1.5 to 100% within individual host species. Phylogenetic analysis of these Cryptosporidium isolates as well as 14 isolates from previous studies indicated extensive genetic diversity as well as evidence for mixed infections. At the 18S locus the following species were identified; Cryptosporidium molnari-like genotype (n=14), Cryptosporidium huwi (n=8), piscine genotype 2 (n=4), piscine genotype 3-like (n=1), piscine genotype 4 (n=2), piscine genotype 5 (n=13), piscine genotype 5-like (n=1) and five novel genotypes (n=5). At the actin locus, species identification agreed with the 18S locus for only 52.3% of isolates sequenced, indicating high levels of mixed infections. Future studies will need to employ both morphological characterization and deep sequencing amplicon-based technologies to better understand the epidemiological and phylogenetic relationships of piscine-derived Cryptosporidium species and genotypes, particularly when mixed infections are detected. PMID:26527238

  12. NADPH oxidase, MPO, NE, ERK1/2, p38 MAPK and Ca2+ influx are essential for Cryptosporidium parvum-induced NET formation.

    PubMed

    Muñoz-Caro, Tamara; Lendner, Matthias; Daugschies, Arwid; Hermosilla, Carlos; Taubert, Anja

    2015-10-01

    Cryptosporidium parvum causes a zoonotic infection with worldwide distribution. Besides humans, cryptosporidiosis affects a wide range of animals leading to significant economic losses due to severe enteritis in neonatal livestock. Neutrophil extracellular trap (NET) formation has been demonstrated as an important host effector mechanism of PMN acting against several invading pathogens. In the present study, C. parvum-mediated NET formation was investigated in human and bovine PMN in vitro. We here demonstrate that C. parvum sporozoites indeed trigger NET formation in a time-dependent manner. Thereby, the classical characteristics of NETs were demonstrated by co-localization of extracellular DNA with histones, neutrophil elastase (NE) and myeloperoxidase (MPO). A significant reduction of NET formation was measured following treatments of PMN with NADPH oxidase-, NE- and MPO-inhibitors, confirming the key role of these enzymes in C. parvum-induced NETs. Additionally, sporozoite-triggered NETosis revealed as dependent on intracellular Ca(++) concentration and the ERK 1/2 and p38 MAPK-mediated signaling pathway. Moreover, sporozoite-triggered NET formation led to significant parasite entrapment since 15% of the parasites were immobilized in NET structures. Consequently, PMN-pre-exposed sporozoites showed significantly reduced infectivity for epithelial host cells confirming the capability of NETs to prevent active parasite invasion. Besides NETs, we here show that C. parvum significantly up-regulated CXCL8, IL6, TNF-α and of GM-CSF gene transcription upon sporozoite confrontation, indicating a pivotal role of PMN not only in the bovine and human system but most probably in other final hosts for C. parvum. PMID:26026247

  13. Prevalence and characterization of Cryptosporidium spp. in dairy cattle in Nile River delta provinces, Egypt.

    PubMed

    Amer, Said; Zidan, Shereif; Adamu, Haileeyesus; Ye, Jianbin; Roellig, Dawn; Xiao, Lihua; Feng, Yaoyu

    2013-11-01

    Molecular characterizations of Cryptosporidium spp. in dairy cattle in industrialized nations have mostly shown a dominance of Cryptosporidium parvum, especially its IIa subtypes in pre-weaned calves. Few studies, however, have been conducted on the distribution of Cryptosporidium species and C. parvum subtypes in various age groups of dairy cattle in developing countries. In this study, we examined the prevalence and molecular characteristics of Cryptosporidium in dairy cattle in four Nile River delta provinces in Egypt. Modified Ziehl-Neelsen acid-fast microscopy was used to screen for Cryptosporidium oocysts in 1974 fecal specimens from animals of different ages on 12 farms. Positive fecal specimens were identified from all studied farms with an overall prevalence of 13.6%. By age group, the infection rates were 12.5% in pre-weaned calves, 10.4% in post-weaned calves, 22.1% in heifers, and 10.7% in adults. PCR-RFLP and DNA sequence analyses of microscopy-positive fecal specimens revealed the presence of four major Cryptosporidium species. In pre-weaned calves, C. parvum was most common (30/69 or 43.5%), but Cryptosporidium ryanae (13/69 or 18.8%), Cryptosporidium bovis (7/69 or 10.2%), and Cryptosporidium andersoni (7/69 or 10.2%) were also present at much higher frequencies seen in most industrialized nations. Mixed infections were seen in 12/69 (17.4%) of genotyped specimens. In contrast, C. andersoni was the dominant species (193/195 or 99.0%) in post-weaned calves and older animals. Subtyping of C. parvum based on sequence analysis of the 60kDa glycoprotein gene showed the presence of subtypes IIdA20G1 in nine specimens, IIaA15G1R1 in 27 specimens, and a rare subtype IIaA14G1R1r1b in one specimen. The common occurrence of non-C. parvum species and IId subtypes in pre-weaned calves is a distinct feature of cryptosporidiosis transmission in dairy cattle in Egypt. The finding of the same two dominant IIa and IId C. parvum subtypes recently found in humans in

  14. First report of Cryptosporidium canis in foxes (Vulpes vulpes) and raccoon dogs (Nyctereutes procyonoides) and identification of several novel subtype families for Cryptosporidium mink genotype in minks (Mustela vison) in China.

    PubMed

    Zhang, Siwen; Tao, Wei; Liu, Chengwu; Jiang, Yanxue; Wan, Qiang; Li, Qiao; Yang, Hang; Lin, Yongchao; Li, Wei

    2016-07-01

    Despite the rapid and extensive advances in molecular epidemiology of Cryptosporidium in humans and a variety of animals, the prevalence and genetic traits of the parasite in wildlife bred in captivity and the role of the neglected hosts in zoonotic transmission of human cryptosporidiosis are rarely understood. This study investigated the prevalence, species/genotype, and subtype of Cryptosporidium in farmed fur animals in China and assessed the possibility of zoonotic transmission. Three of 191 (1.6%) foxes (Vulpes vulpes), 17 of 162 (10.5%) raccoon dogs (Nyctereutes procyonoides), and 48 of 162 (29.6%) minks (Mustela vison) were positive for Cryptosporidium by nested PCRs targeting the small subunit rRNA gene. Sequence analysis indicated the presence of only Cryptosporidium canis in foxes and raccoon dogs. There is no significant difference in prevalence between young and adult foxes (or raccoon dogs). Three Cryptosporidium species or genotype including C. canis, Cryptosporidium meleagridis, and mink genotype were determined in minks aged five to six months. Subtyping based on nucleotide and amino acid sequence polymorphisms of the 60kDa glycoprotein facilitated identification of three novel subtype families named as Xb to Xd for Cryptosporidium mink genotype. The presence of zoonotic C. canis, C. meleagridis, and Cryptosporidium mink genotype in captive-bred fur animals is of public health concerns. The findings expanded the host ranges of C. canis and C. meleagridis and confirmed genetic diversity at the subtype level in Cryptosporidium mink genotype. This is the first study reporting Cryptosporidium infections in foxes and raccoon dogs in China. PMID:27001467

  15. Occurrence of Cryptosporidium oocysts in fecal samples submitted for routine microbiological examination.

    PubMed Central

    Ratnam, S; Paddock, J; McDonald, E; Whitty, D; Jong, M; Cooper, R

    1985-01-01

    During a 7-month period, 2,252 fecal samples submitted for routine microbiological examination from 1,621 patients were screened for Cryptosporidium oocysts by the auramine staining method with Kinyoun acid-fast stain as the confirmatory stain. Cryptosporidium oocysts were detected in fecal samples from 19 (1.2%) patients, 18 of whom had gastroenteritis. Diarrheic stools from 14 of these 18 patients were negative for the usual enteropathogens but contained the oocysts in moderate to large numbers. Although Cryptosporidium oocysts were found in patients of all ages, they occurred slightly more frequently in infants and children than in the rest. Cryptosporidium species was one of the common enteropathogens identified in fecal samples submitted for routine parasitological examination during the period of the survey and was second only to Giardia species in terms of frequency. Considering cryptosporidiosis in the differential diagnosis of gastroenteritis in immunocompetent persons and including a search for Cryptosporidium oocysts in routine parasitological examinations of fecal samples appear warranted. PMID:4044798

  16. Cryptosporidium scrofarum n. sp. (Apicomplexa: Cryptosporidiidae) in domestic pigs (Sus scrofa)

    PubMed Central

    Kváč, Martin; Kestřánová, Michaela; Pinková, Martina; Květoňová, Dana; Kalinová, Jana; Wagnerová, Pavla; Kotková, Michaela; Vítovec, Jiří; Ditrich, Oleg; McEvoy, John; Stenger, Brianna; Sak, Bohumil

    2012-01-01

    We describe the morphological, biological, and molecular characteristics of Cryptosporidium pig genotype II and propose the species name Cryptosporidium scrofarum n. sp. to reflect its prevalence in adult pigs worldwide. Oocysts of C. scrofarum are morphologically indistinguishable from C. parvum, measuring 4.81–5.96 µm (mean = 5.16) × 4.23–5.29 µm (mean = 4.83) with a length to width ratio of 1.07 ± 0.06 (n = 400). Oocysts of C. scrofarum obtained from a naturally infected pig were infectious for 8-week-old pigs but not 4-week-old pigs. The prepatent period in 8-week-old Cryptosporidium-naive pigs was 4–6 days and the patent period was longer than 30 days. The infection intensity of C. scrofarum in pigs was generally low, in the range 250-4000 oocysts per gram of faeces. Infected pigs showed no clinical signs of cryptosporidiosis and no pathology was detected. Cryptosporidium scrofarum was not infectious for adult SCID mice, adult BALB c mice, Mongolian gerbils (Meriones unguiculatus), southern multimammate mice (Mastomys coucha), yellow-necked mice (Apodemus flavicollis), or guinea pigs (Cavia porcellus). Phylogenetic analyses based on Small subunit rRNA, actin, and heat shock protein 70 gene sequences revealed that C. scrofarum is genetically distinct from all known Cryptosporidium species. PMID:23021264

  17. Molecular characterization of Cryptosporidium isolates from pigs in Zaragoza (northeastern Spain).

    PubMed

    Suárez-Luengas, Laura; Clavel, Antonio; Quílez, Joaquín; Goñi-Cepero, María Pilar; Torres, Eucaris; Sánchez-Acedo, Caridad; del Cacho, Emilio

    2007-09-30

    A total of 142 stool specimens from pigs on 24 farms from the province of Zaragoza (northeastern Spain) were screened for Cryptosporidium spp. Samples were first analysed by routine techniques (formalin-ethyl acetate sedimentation method and modified Ziehl-Neelsen stain) selecting those microscopically positive for genetic characterization. Cryptosporidium species and genotypes were determined by a nested PCR-RFLP technique at the 18S ribosomal DNA locus and sequencing of the PCR-positive secondary products. Cryptosporidium oocysts were microscopically identified in the faeces of 32 pigs (22.5%) from 15 farms (62.5%). Infected animals included 23 weaned piglets (30.7%), 5 fattening pigs (11.9%) and 4 sows (16%). Diarrhoea was not detected in any of the infected pigs. The molecular characterization was successfully performed in 26 samples from 14 farms. Cryptosporidium suis was found in 10 specimens from 7 farms (nine weaned piglets and one sow) and the Cryptosporidium pig genotype II in 16 samples from 10 farms (13 weaned piglets and 3 fattening pigs). Both C. suis and the pig genotype II were concurrently detected on three farms. PMID:17683866

  18. Prevalence of Cryptosporidium genotypes in pre and post-weaned pigs in Australia.

    PubMed

    Johnson, Johanna; Buddle, Ross; Reid, Simon; Armson, Anthony; Ryan, Una M

    2008-07-01

    A total of 289 pig faecal samples were collected from pre-weaned and post-weaned piglets and sows from 1 indoor and 3 outdoor piggeries located in the south-west region of Western Australia and screened at the 18S rRNA locus for the presence of Cryptosporidium. An overall prevalence of 22.1% (64/289) was identified. Cryptosporidium was more prevalent in post-weaned animals (p<0.05); 32.7% (51/156) versus 10.6% (13/123) for pre-weaned animals. Sequence analysis identified Cryptosporidium suis in all pre-weaned isolates genotyped (7/13). In post-weaned pigs that were genotyped (n=38), the non-zoonotic Cryptosporidium species, pig genotype II was identified in 32 samples and C. suis in 6 samples. The zoonotic species Cryptosporidium parvum was not detected, suggesting that domestic pigs do not pose a significant public health risk. Pig genotype II was significantly (p<0.05) associated with 'normal' stools, indicating an asymptomatic nature in the porcine host. PMID:18486131

  19. Prevalence and pathogenicity of Cryptosporidium suis in pre- and post-weaned pigs.

    PubMed

    Vítovec, J; Hamadejová, K; Landová, L; Kvác, M; Kvetonová, D; Sak, B

    2006-06-01

    A total of 4338 faecal samples, 135 of sows, 3368 of pre-weaned and 835 of post-weaned piglets from eight farms in South Bohemia, Czech Republic were collected and examined for Cryptosporidium infection. No sow, but 5.7% pre-weaned and 24.1% post-weaned piglets were positive for Cryptosporidium infection. No relationship was found between diarrhoea and Cryptosporidium infection in any of the different age groups (pre- and post-weaned piglets). Four piglets, which were sporadically shedding cryptosporidia in faeces, were necropsied. Neither clinical signs of diarrhoea nor macroscopical changes were found. Histologically, a moderate infection of cryptosporidia was detected in the glandular epithelium along the large intestine, with predisposition to the ansa centralis of the colon. No inflammatory response in the lamina propria was observed. Cryptosporidia were also commonly found in the glandular epithelium of submucosal lymphoglandular complexes in the colon. Cryptosporidium isolates from all farms were identified as Cryptosporidium suis using molecular markers (SSU rRNA). All of the C. suis strains obtained were larger [6.2 (6.0-6.8) x 5.5 (5.3-5.7) microm] than any isolate described so far [4.6 (4.4-4.9) x 4.2 (4.0-4.3) microm] and did not appear to be infective for neonatal BALB/c mice. PMID:16732883

  20. Cryptosporidium within-host genetic diversity: systematic bibliographical search and narrative overview.

    PubMed

    Grinberg, Alex; Widmer, Giovanni

    2016-07-01

    Knowledge of the within-host genetic diversity of a pathogen often has broad implications for disease management. Cryptosporidium protozoan parasites are among the most common causative agents of infectious diarrhoea. Current limitations of in vitro culture impose the use of uncultured isolates obtained directly from the hosts as operational units of Cryptosporidium genotyping. The validity of this practice is centred on the assumption of genetic homogeneity of the parasite within the host, and genetic studies often take little account of the within-host genetic diversity of Cryptosporidium. Yet, theory and experimental evidence contemplate genetic diversity of Cryptosporidium at the within-host scale, but this diversity is not easily identified by genotyping methods ill-suited for the resolution of DNA mixtures. We performed a systematic bibliographical search of the occurrence of within-host genetic diversity of Cryptosporidium parasites in epidemiological samples, between 2005 and 2015. Our results indicate that genetic diversity at the within-host scale, in the form of mixed species or intra-species diversity, has been identified in a large number (n=55) of epidemiological surveys of cryptosporidiosis in variable proportions, but has often been treated as a secondary finding and not analysed. As in malaria, there are indications that the scale of this diversity varies between geographical regions, perhaps depending on the prevailing transmission pathways. These results provide a significant knowledge base from which to draw alternative population genetic structure models, some of which are discussed in this paper. PMID:27021167

  1. Cryptosporidium scrofarum n. sp. (Apicomplexa: Cryptosporidiidae) in domestic pigs (Sus scrofa).

    PubMed

    Kváč, Martin; Kestřánová, Michaela; Pinková, Martina; Květoňová, Dana; Kalinová, Jana; Wagnerová, Pavla; Kotková, Michaela; Vítovec, Jiří; Ditrich, Oleg; McEvoy, John; Stenger, Brianna; Sak, Bohumil

    2013-01-31

    We describe the morphological, biological, and molecular characteristics of Cryptosporidium pig genotype II and propose the species name Cryptosporidium scrofarum n. sp. to reflect its prevalence in adult pigs worldwide. Oocysts of C. scrofarum are morphologically indistinguishable from C. parvum, measuring 4.81-5.96 μm (mean=5.16)×4.23-5.29 μm (mean=4.83) with a length to width ratio of 1.07±0.06 (n=400). Oocysts of C. scrofarum obtained from a naturally infected pig were infectious for 8-week-old pigs but not 4-week-old pigs. The prepatent period in 8-week-old Cryptosporidium-naive pigs was 4-6 days and the patent period was longer than 30 days. The infection intensity of C. scrofarum in pigs was generally low, in the range 250-4000 oocysts per gram of feces. Infected pigs showed no clinical signs of cryptosporidiosis and no pathology was detected. Cryptosporidium scrofarum was not infectious for adult SCID mice, adult BALB/c mice, Mongolian gerbils (Meriones unguiculatus), southern multimammate mice (Mastomys coucha), yellow-necked mice (Apodemus flavicollis), or guinea pigs (Cavia porcellus). Phylogenetic analyses based on small subunit rRNA, actin, and heat shock protein 70 gene sequences revealed that C. scrofarum is genetically distinct from all known Cryptosporidium species. PMID:23021264

  2. Removal of Cryptosporidium and polystyrene microspheres from swimming pool water with sand, cartridge, and precoat filters.

    PubMed

    Amburgey, James E; Walsh, Kimberly J; Fielding, Roy R; Arrowood, Michael J

    2012-03-01

    Cryptosporidium has caused the majority of waterborne disease outbreaks in treated recreational water venues in the USA for many years running. This research project evaluated some common US swimming pool filters for removing Cryptosporidium oocysts, 5-µm diameter polystyrene microspheres, and 1-µm diameter polystyrene microspheres. A 946 L hot tub with interchangeable sand, cartridge, and precoat filters was used at room temperature for this research. Simulated pool water for each experiment was created from Charlotte, NC (USA) tap water supplemented with alkalinity, hardness, chlorine, and a mixture of artificial sweat and urine. Precoat (i.e., diatomaceous earth and perlite) filters demonstrated pathogen removal efficiencies of 2.3 to 4.4 log (or 99.4-99.996%). However, sand and cartridge filters had average Cryptosporidium removals of 0.19 log (36%) or less. The combined low filter removal efficiencies of sand and cartridge filters along with the chlorine-resistant properties of Cryptosporidium oocysts could indicate a regulatory gap warranting further attention and having significant implications on the protection of public health in recreational water facilities. The 5-µm microspheres were a good surrogate for Cryptosporidium oocysts in this study and hold promise for use in future research projects, field trials, and/or product testing on swimming pool filters. PMID:22361700

  3. Giardia and Cryptosporidium in harp and hooded seals from the Gulf of St. Lawrence, Canada.

    PubMed

    Appelbee, A J; Thompson, R C A; Measures, L M; Olson, M E

    2010-10-11

    Giardia and Cryptosporidium are protozoan parasites known to cause enteric disease in terrestrial wildlife species (mammals, reptiles and birds). Few surveys for Giardia and Cryptosporidium in marine wildlife species, such as pinnipeds, have been reported. The objective of this study was to determine the prevalence and genotype of Giardia and Cryptosporidium in two species of pinnipeds, harp seal (Phoca groenlandica) and hooded seal (Cystophora cristata), from the Gulf of St. Lawrence, Canada. Faecal samples were collected from pup and adult seals and examined for the presence of cysts of Giardia and oocysts of Cryptosporidium using microscopy and immunofluorescent staining. Tissues from the small intestine of adult seals were also collected and examined for infections using the polymerase chain reaction (PCR) technique. Giardia cysts were found in the faeces of 42% (16/38) of adult harp seals, but in none of the harp seal pups (0/20). Although Giardia cysts were not detected in faeces of adult hooded seals (0/10) using microscopy, 80% tested positive for Giardia using PCR of intestinal tissue indicative of a true replicating infection. Both harp and hooded seals harboured infections with the zoonotic strain, Giardia duodenalis Assemblage A, as determined using a nested-PCR technique to amplify a small subunit ribosomal (SSU-rRNA) gene of Giardia. Cryptosporidium was not detected by microscopy, nor using the PCR technique on intestinal tissues from any of the 68 seals examined. PMID:20594649

  4. Prevalence of Cryptosporidium Infection among Inhabitants of 2 Rural Areas in White Nile State, Sudan

    PubMed Central

    Sim, Seobo; Yu, Jae-Ran; Lee, Young-Ha; Lee, Jin-Su; Jeong, Hoo-Gn; Mohamed, Abd Al Wahab Saed; Hong, Sung-Tae

    2015-01-01

    Cryptosporidium , a protozoan parasite that causes watery diarrhea, is found worldwide and is common in areas with low water hygiene. In February 2014, 866 stool samples were collected from the inhabitants of 2 rural areas in White Nile State, Sudan. These stool samples were assessed by performing modified acid-fast staining, followed by examination under a light microscope. The overall positive rate of Cryptosporidium oocysts was 13.3%. Cryptosporidium oocysts were detected in 8.6% stool samples obtained from inhabitants living in the area having water purification systems and in 14.6% stool samples obtained from inhabitants living in the area not having water purification systems. No significant difference was observed in the prevalence of Cryptosporidium infection between men and women (14.7% and 14.1%, respectively). The positive rate of oocysts by age was the highest among inhabitants in their 60s (40.0%). These findings suggest that the use of water purification systems is important for preventing Cryptosporidium infection among inhabitants of these rural areas in Sudan. PMID:26797443

  5. Efficacy of Nitazoxanide against Cryptosporidium parvum in Cell Culture and in Animal Models

    PubMed Central

    Theodos, Cynthia M.; Griffiths, Jeffrey K.; D’Onfro, Jennifer; Fairfield, Alexandra; Tzipori, Saul

    1998-01-01

    Nitazoxanide (NTZ), a drug currently being tested in human clinical trials for efficacy against chronic cryptosporidiosis, was assessed in cell culture and in two animal models. The inhibitory activity of NTZ was compared with that of paromomycin (PRM), a drug that is partially effective against Cryptosporidium parvum. A concentration of 10 μg of NTZ/ml (32 μM) consistently reduced parasite growth in cell culture by more than 90% with little evidence of drug-associated cytotoxicity, in contrast to an 80% reduction produced by PRM at 2,000 μg/ml (3.2 mM). In contrast to its efficacy in vitro, NTZ at either 100 or 200 mg/kg of body weight/day for 10 days was ineffective at reducing the parasite burden in C. parvum-infected, anti-gamma-interferon-conditioned SCID mice. Combined treatment with NTZ and PRM was no more effective than treatment with PRM alone. Finally, NTZ was partially effective at reducing the parasite burden in a gnotobiotic piglet diarrhea model when given orally for 11 days at 250 mg/kg/day but not at 125 mg/kg/day. However, the higher dose of NTZ induced a drug-related diarrhea in piglets that might have influenced its therapeutic efficacy. As we have previously reported, PRM was effective at markedly reducing the parasite burden in piglets at a dosage of 500 mg/kg/day. Our results indicate that of all of the models tested, the piglet diarrhea model most closely mimics the partial response to NTZ treatment reported to occur in patients with chronic cryptosporidiosis. PMID:9687390

  6. 78 FR 14508 - Notice of Affirmation of Addition of a Treatment Schedule for Methyl Bromide Fumigation of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-06

    ... Treatment Schedule for Methyl Bromide Fumigation of Cottonseed AGENCY: Animal and Plant Health Inspection... methyl bromide fumigation of cottonseed for the fungal plant pathogen Fusarium oxysporum f. sp... Manual of the treatment described in the notice published at 77 FR 31564-31566 on May 29, 2012....

  7. Removal effect of the water purifier for home use against Cryptosporidium parvum oocysts.

    PubMed

    Matsui, Toshihiro; Kajima, Junko; Fujino, Takashi

    2004-08-01

    The removal effects of the faucet mounted type water purifier for home use were examined against Cryptosporidium parvum oocysts. The water purifier is composed of a layer of granular activated carbon and the hollow fiber membrane filter. The cartridges were unused, 25%, 50% and 75% flow down by Arizona-dust of U. S. A. Two respective cartridges were used of the examination. The faucet and the water purifier were connected by anti-pressure tube, and 3.0 x 10(7) oocysts of Cryptosporidium parvum were injected into anti-pressure tube while water was running. Twenty liter of collected purified water was examined under the fluorescent microscope. Any oocysts in the purified water collected from all cartridges were not found. Therefore, we considered this purifier as an effective one in removing Cryptosporidium oocysts from drinking water. PMID:15353844

  8. [Development of a technique for recovering Giardia cysts and Cryptosporidium oocysts from fresh vegetables].

    PubMed

    Di Benedetto, M A; Di Piazza, F; Oliveri, R; Cerame, G; Valenti, R; Firenze, A

    2006-01-01

    Techniques described for recovering Giardia and Cryptosporidium (oo)cysts from fruit and vegetables are generally inadequate and present variable recovery efficience and elevated costs. The aim of our study was to evaluate the recovery efficiency of a simple and economic technique to apply either to berry vegetables, like tomatoes and peppers, or to large leave vegetables, like lettuce and chicory. The method include contamination and further elution of the vegetables. Then sedimentation of (oo)cysts by centrifugation of the eluate of vegetables and their visualization by means of direct immunofluorescence. The higher recovery values for both protozoa were obtained in large leave vegetables with mean data above 70% for Giardia and 76% for Cryptosporidium, whereas the values observed in the berry vegetables were above 43% for Giardia and above 37% for Cryptosporidium on average. PMID:16649507

  9. Comparison of selected methods for recovery of Giardia spp. cysts and Cryptosporidium spp. oocysts in wastewater.

    PubMed

    Medeiros, Raphael Corrêa; Daniel, Luiz Antonio

    2015-09-01

    More precise methods are needed to recover Giardia and Cryptosporidium (oo)cysts from wastewater in order to advance research related to their inactivation, removal, quantification, and species differentiation. This study applied different methods to recover the maximum number of (oo)cysts from wastewater samples using ColorSeed®. Immunomagnetic separation assisted in capturing oocysts mainly in samples with medium and low turbidity. A triple centrifugation method reached recovery rates of 85% and 20%, for Giardia cysts and Cryptosporidium oocysts, respectively, in raw wastewater, and 62.5 and 17.5% in secondary-treated effluent. For low turbidity-treated effluent, membrane filtration reached 67.5% recovery for Giardia cysts and 22.5% for Cryptosporidium oocysts. Simple, quick and low-cost methods do not involve much handling of the samples and could be useful, particularly in developing countries. PMID:26322766

  10. Giardia spp. and Cryptosporidium spp. In the Ivaí Indigenous Land, Brazil.

    PubMed

    Nishi, Letícia; Bergamasco, Rosângela; Toledo, Max Jean de Ornelas; Falavigna, Dina Lúcia Morais; Gomes, Mônica Lúcia; Mota, Lúcio Tadeu; Falavigna-Guilherme, Ana Lúcia

    2009-10-01

    The objective of this study was to investigate the occurrence of cysts of Giardia spp. and oocysts of Cryptosporidium spp. in waters of the Ivaí Indigenous Land, Brazil. Samples of river and spring water and of treated water were filtered and analyzed by direct immunofluorescence (Merifluor kit, Meridian Bioscience, Cincinnati, Ohio). Of 21 samples, 7 from each locality, 3 (3/7, 42.8%) from a river were positive for Giardia (mean concentration 2.57 cysts/L), and 1 (1/7, 14.3%) was positive for Cryptosporidium (6 oocysts/L). From springs, 1 sample (1/7, 14.3%) was positive for Cryptosporidium (6 oocysts/L). One sample (1/7, 14.3%) from treated water was positive for both, with 4 oocysts/L and 2 cysts/L. Giardia was the more frequent protozoan present. PMID:18945186

  11. Common occurrence of zoonotic pathogen Cryptosporidium meleagridis in broiler chickens and turkeys in Algeria.

    PubMed

    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. PMID:23498647

  12. Development and Application of a gp60-Based Typing Assay for Cryptosporidium viatorum

    PubMed Central

    Elwin, K.; Winiecka-Krusnell, J.; Chalmers, R. M.; Xiao, L.; Lebbad, M.

    2015-01-01

    The apicomplexan intestinal parasites of the genus Cryptosporidium take a major toll on human and animal health and are frequent causes of waterborne outbreaks. Several species and genotypes can infect humans, including Cryptosporidium viatorum, which, to date, has only been found in humans. Molecular characterization of Cryptosporidium spp., critical to epidemiological analyses, is commonly based on gp60 gene analysis, which appears to require bespoke species- or group-specific PCR primers due to extensive genetic diversity across the genus. In this study, we amplified, sequenced, and characterized the gp60 gene of C. viatorum for the first time. Moreover, we developed and validated a gp60 typing assay for this species and applied it to 27 isolates originating from Asia, Africa, and Central America. A single subtype family, XVa, was identified containing multiple alleles. PMID:25832304

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

    USGS Publications Warehouse

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

    2006-01-01

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

  14. Expression of Cpgp40/15 in Toxoplasma gondii: a surrogate system for the study of Cryptosporidium glycoprotein antigens.

    PubMed

    O'Connor, R M; Kim, K; Khan, F; Ward, H D

    2003-10-01

    Cryptosporidium parvum is a waterborne enteric coccidian that causes diarrheal disease in a wide range of hosts. Development of successful therapies is hampered by the inability to culture the parasite and the lack of a transfection system for genetic manipulation. The glycoprotein products of the Cpgp40/15 gene, gp40 and gp15, are involved in C. parvum sporozoite attachment to and invasion of host cells and, as such, may be good targets for anticryptosporidial therapies. However, the function of these antigens appears to be dependent on the presence of multiple O-linked alpha-N-acetylgalactosamine (alpha-GalNAc) determinants. A eukaryotic expression system that would produce proteins bearing glycosylation patterns similar to those found on the native C. parvum glycoproteins would greatly facilitate the molecular and functional characterization of these antigens. As a unique approach to this problem, the Cpgp40/15 gene was transiently expressed in Toxoplasma gondii, and the expressed recombinant glycoproteins were characterized. Antisera to gp40 and gp15 reacted with the surface membranes of tachyzoites expressing the Cpgp40/15 construct, and this reactivity colocalized with that of antiserum to the T. gondii surface protein SAG1. Surface membrane localization was dependent on the presence of the glycophosphatidylinositol anchor attachment site present in the gp15 coding sequence. The presence of terminal O-linked alpha-GalNAc determinants on the T. gondii recombinant gp40 was confirmed by reactivity with Helix pomatia lectin and the monoclonal antibody 4E9, which recognizes alpha-GalNAc residues, and digestion with alpha-N-acetylgalactosaminidase. In addition to appropriate localization and glycosylation, T. gondii apparently processes the gp40/15 precursor into the gp40 and gp15 component glycopolypeptides, albeit inefficiently. These results suggest that a surrogate system using T. gondii for the study of Cryptosporidium biology may be useful. PMID:14500524

  15. MicroRNA-221 controls expression of intercellular adhesion molecule-1 in epithelial cells in response to Cryptosporidium parvum infection

    PubMed Central

    Gong, Ai-Yu; Hu, Guoku; Zhou, Rui; Liu, Jun; Feng, Yaoyu; Soukup, Garrett A.; Chen, Xian-Ming

    2011-01-01

    Cryptosporidium parvum is a protozoan parasite that infects gastrointestinal epithelial cells and causes diarrheal disease in humans and animals globally. Pathological changes following C. parvum infection include crypt hyperplasia, a modest inflammatory reaction with increased infiltration of lymphocytes into intestinal mucosa. Expression of adhesion molecules such as intercellular adhesion molecule-1 (ICAM-1), on infected epithelial cell surfaces may facilitate adhesion and recognition of lymphocytes at infection sites. MicroRNAs (miRNAs) are small RNA molecules of 23 nucleotides that negatively regulate protein-coding gene expression via translational suppression or mRNA degradation. We recently reported that microRNA-221 (miR-221) regulates ICAM-1 translation through targeting the ICAM-1 3′-untranslated region (UTR). In this study, we tested the role of miR-221 in regulating ICAM-1 expression in epithelial cells in response to C. parvum infection using an in vitro model of human biliary cryptosporidiosis. Up-regulation of ICAM-1 at both message and protein levels was detected in epithelial cells following C. parvum infection. Inhibition of ICAM-1 transcription with actinomycin D could only partially block C. parvum-induced ICAM-1 expression at the protein level. Cryptosporidium parvum infection decreased miR-221 expression in infected epithelial cells. When cells were transfected with a luciferase reporter construct covering the miR-221 binding site in the ICAM-1 3′-UTR and then exposed to C. parvum, an enhanced luciferase activity was detected. Transfection of miR-221 precursor abolished C. parvum-stimulated ICAM-1 protein expression. In addition, expression of ICAM-1 on infected epithelial cells facilitated epithelial adherence of co-cultured Jurkat cells. These results indicate that miR-221-mediated translational suppression controls ICAM-1 expression in epithelial cells in response to C. parvum infection. PMID:21236259

  16. Highly specific detection of Cryptosporidium spp. oocysts in human stool samples by undemanding and inexpensive phase contrast microscopy.

    PubMed

    Ignatius, Ralf; Klemm, Thomas; Zander, Steffen; Gahutu, Jean Bosco; Kimmig, Peter; Mockenhaupt, Frank P; Regnath, Thomas

    2016-03-01

    To compare phase contrast microscopy (PCM) of unstained slides for the detection of Cryptosporidium spp. oocysts with a commercially available enzyme immunoassay (EIA) for the detection of cryptosporidial antigen in human stool samples, we prospectively analysed by both methods 463 fresh human stool samples obtained from diarrhoeic patients between July and October 2014. Compared with the EIA, the sensitivity, specificity, positive and negative predictive value of PCM were 88.9 % (95 % confidence interval (CI), 66.0-98.1 %), 100 % (95 % CI, 99.0-100 %), 100 % (95 % CI, 77.3-100 %) and 99.6 % (95 % CI, 98.3-100 %), respectively. Additionally, we retrospectively examined with PCM 65 fixed stool samples that had been collected in 2010 from mostly asymptomatic Rwandan children <5 years of age; 14 of these samples had previously yielded positive results with a highly sensitive real-time (RT)-PCR. PCM detected cryptosporidia in 5/14 RT-PCR-positive samples, and notably, also in one of 51 RT-PCR-negative samples, which was subsequently confirmed by acid-fast staining. Positive and negative percent agreement of PCM with RT-PCR were 35.7 % (95 % CI, 16.2-61.4 %) and 98.0 % (95 % CI, 88.7-100 %), respectively. Positive PCM results were associated with higher RT-PCR cycle threshold values (p = 0.044). In conclusion, PCM offers a highly specific, undemanding and inexpensive method for the laboratory diagnosis of acute human cryptosporidiosis independent of the causative Cryptosporidium species. PMID:26646397

  17. 49 CFR 40.303 - What happens if the SAP believes the employee needs additional treatment, aftercare, or support...

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 1 2012-10-01 2012-10-01 false What happens if the SAP believes the employee... the Return-to-Duty Process § 40.303 What happens if the SAP believes the employee needs additional...? (a) As a SAP, if you believe that ongoing services (in addition to follow-up tests) are needed...

  18. 49 CFR 40.303 - What happens if the SAP believes the employee needs additional treatment, aftercare, or support...

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 1 2013-10-01 2013-10-01 false What happens if the SAP believes the employee... the Return-to-Duty Process § 40.303 What happens if the SAP believes the employee needs additional...? (a) As a SAP, if you believe that ongoing services (in addition to follow-up tests) are needed...

  19. 49 CFR 40.303 - What happens if the SAP believes the employee needs additional treatment, aftercare, or support...

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 1 2010-10-01 2010-10-01 false What happens if the SAP believes the employee... the Return-to-Duty Process § 40.303 What happens if the SAP believes the employee needs additional...? (a) As a SAP, if you believe that ongoing services (in addition to follow-up tests) are needed...

  20. 49 CFR 40.303 - What happens if the SAP believes the employee needs additional treatment, aftercare, or support...

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 1 2011-10-01 2011-10-01 false What happens if the SAP believes the employee... the Return-to-Duty Process § 40.303 What happens if the SAP believes the employee needs additional...? (a) As a SAP, if you believe that ongoing services (in addition to follow-up tests) are needed...