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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. Bin 2 1-log treatment 1.5-log treatment 1-log treatment (1) Bin 3 2-log treatment 2.5-log treatment 2-log treatment (2) Bin 4 2.5-log treatment 3-log treatment 2.5-log treatment (3) 1 As determined by the State such that the total Cryptosporidium removal and inactivation is at least 4.0-log. 2...

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

    Code of Federal Regulations, 2012 CFR

    2012-07-01

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

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

    Code of Federal Regulations, 2013 CFR

    2013-07-01

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

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

    Code of Federal Regulations, 2011 CFR

    2011-07-01

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

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

    Code of Federal Regulations, 2014 CFR

    2014-07-01

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

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

    PubMed

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

    2015-01-01

    Wastewater disposal may be a source of environmental contamination by Cryptosporidium and Giardia. This study was conducted to evaluate the prevalence of Cryptosporidium oocysts and Giardia cysts in raw and treated wastewater effluents. A prevalence of 100% was demonstrated for Giardia cysts in raw wastewater, at a concentration range of 10 to 12,225 cysts L(-1), whereas the concentration of Cryptosporidium oocysts in raw wastewater was 4 to 125 oocysts L(-1). The removal of Giardia cysts by secondary and tertiary treatment processes was greater than those observed for Cryptosporidium oocysts and turbidity. Cryptosporidium and Giardia were present in 68.5% and 76% of the tertiary effluent samples, respectively, at an average concentration of 0.93 cysts L(-1) and 9.94 oocysts L(-1). A higher detection limit of Cryptosporidium oocysts in wastewater was observed for nested PCR as compared to immune fluorescent assay (IFA). C. hominis was found to be the dominant genotype in wastewater effluents followed by C. parvum and C. andersoni or C. muris. Giardia was more prevalent than Cryptosporidium in the studied community and treatment processes were more efficient for the removal of Giardia than Cryptosporidium. Zoonotic genotypes of Cryptosporidium were also present in the human community. To assess the public health significance of Cryptosporidium oocysts present in tertiary effluent, viability (infectivity) needs to be assessed. PMID:26301853

  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.

  10. 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

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

    PubMed

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

    2007-06-01

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

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

    Code of Federal Regulations, 2013 CFR

    2013-07-01

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

  13. 40 CFR 141.712 - Unfiltered system Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

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

  14. 40 CFR 141.712 - Unfiltered system Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

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

  15. 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...

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

    PubMed

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

    2006-10-01

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

  17. 40 CFR 141.713 - Schedule for compliance with Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

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

  18. 40 CFR 141.713 - Schedule for compliance with Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

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

  19. 40 CFR 141.713 - Schedule for compliance with Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

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

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

    Code of Federal Regulations, 2014 CFR

    2014-07-01

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

  1. 40 CFR 141.713 - Schedule for compliance with Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

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

  2. 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

  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. [Removal of Cryptosporidium and Giardia in drinking water treatment in a Tuscan area].

    PubMed

    Sacco, C; Bianchi, M; Lorini, C; Burrini, D; Berchielli, S; Lanciotti, E

    2006-01-01

    Five drinking water treatment plants in the Tuscan area, differing in relation to the treatments employed, were monitored for the presence of Cryptosporidium and Giardia (oo)cysts in order to estimate their removal capacity of each works. Twenty-eight water samples (16 from inflow raw water and 12 from outflow drinking water) were analysed during the one-year survey. The US EPA 1623 method was used, obtaining, according to the PHLS quality control system, a recovery rate of 51.3 +/- 22.9% for Cryptosporidium oocysts and 48.8 +/- 20.7% for Giardia cysts. The results showed that both the protozoa were present in surface raw waters: Cryptosporidium in 62.5% and Giardia in 87.5% of the samples. Considering the outflow drinking waters, the removal capacity varied from 100% in the technologically more complex plant (which supplies most of the area), to at least 90% in the remaining four plants, where less complex treatments are carried out. The occurrence of (oo)cysts was not associated with seasonality, turbidity or Clostridium perfringens (Cl. perfringens seems to be related with Cryptosporidium oocysts in surface raw waters; in this respect, our legislation states that the protozoa have to be searched for whenever a water sample is positive for Cl. perfringens). We confirmed the validity of the US-EPA 1623 method in assessing and managing the risk associated with waterborne protozoa, thus a valid tool to safeguard the population from exposure to such pathogens.

  5. 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.

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

    PubMed

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

    2012-11-01

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

  7. Removal and fate of Cryptosporidium in dissolved air drinking water treatment plants.

    PubMed

    Edzwald, J K; Tobiason, J E; Dunn, H; Kaminski, G; Galant, P

    2001-01-01

    In the first part of the paper, data from pilot plant studies are used to evaluate Cryptosporidium removal by dissolved air flotation (DAF) clarification and dual media filters under challenge conditions. Oocyst removals were investigated for design detention times and hydraulic loadings for winter and spring seasons. Coagulation was optimized for turbidity and removal of natural organic matter. DAF performance was better for spring water temperatures achieving 2.5 +/- 0.3 log removal of oocysts compared to 1.7 +/- 0.3 log removal in the winter. Cumulative log removal across DAF and filtration exceeded 5.4, and was not affected by water temperature. Low turbidities and particle counts are indicators of good treatment and good removals of Cryptosporidium. The second part of the paper uses a mathematical model to predict the fate of Cryptosporidium through a DAF plant and the impact of filter backwash recycle on oocyst build-up in the plant influent. Model predictions show that the fate of Cryptosporidium and the build-up of oocysts in the plant influent depend on: DAF performance, the percent of filtered water production used for backwashing, and the percent of filter backwash recycle flow. A DAF plant with 2.5% filtered water production for backwashing and that achieves 1.6 log removal or greater of oocysts by DAF clarification will not have a build-up of oocysts in the plant influent regardless of the recycle rate. This is because the oocysts are concentrated in the DAF floated sludge and not within granular filters.

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

    SciTech Connect

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

    1996-11-01

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

  9. 40 CFR 141.712 - Unfiltered system Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... inactivation. (c) Inactivation treatment technology requirements. Unfiltered systems must use chlorine dioxide... section. (1) Systems that use chlorine dioxide or ozone and fail to achieve the...

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

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... inactivation. (c) Inactivation treatment technology requirements. Unfiltered systems must use chlorine dioxide... section. (1) Systems that use chlorine dioxide or ozone and fail to achieve the...

  11. 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

  12. Treatment of Cryptosporidium: What We Know, Gaps, and the Way Forward

    PubMed Central

    Sparks, Hayley; Nair, Gayatri; Castellanos-Gonzalez, Alejandro; White, A. Clinton

    2015-01-01

    Cryptosporidiosis is increasingly recognized as an important global health concern. While initially reported in immunocompromised such as AIDS patients, cryptosporidiosis has now been documented as a major cause of childhood diarrhea and an important factor in childhood malnutrition. Currently, nitazoxanide is the only proven anti-parasitic treatment for Cryptosporidium infections. However, it is not effective in severely immunocompromised patients and there is limited data in infants. Immune reconstitution or decreased immunosuppression is critical to therapy in AIDS and transplant patients. This limitation of treatment options presents a major public health challenge given the important burden of disease. Repurposing of drugs developed for other indications and development of inhibitors for novel targets offer hope for improved therapies, but none have advanced to clinical studies. PMID:26568906

  13. 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

  14. 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 ...

  15. 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...

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

    PubMed

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

    2007-01-01

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

  17. 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

  18. New developments in Cryptosporidium research.

    PubMed

    Ryan, Una; Hijjawi, Nawal

    2015-05-01

    Cryptosporidium is an enteric parasite that is considered the second greatest cause of diarrhoea and death in children after rotavirus. Currently, 27 species are recognised as valid and of these, Cryptosporidium hominis and Cryptosporidium parvum are responsible for the majority of infections in humans. Molecular and biological studies indicate that Cryptosporidium is more closely related to gregarine parasites rather than to coccidians. The identification of gregarine-like gamont stages and the ability of Cryptosporidium to complete its life cycle in the absence of host cells further confirm its relationship with gregarines. This opens new avenues into the investigation of pathogenesis, epidemiology, treatment and control of Cryptosporidium. Effective drug treatments and vaccines are not yet available due, in part, to the technical challenges of working on Cryptosporidium in the laboratory. Whole genome sequencing and metabolomics have expanded our understanding of the biochemical requirements of this organism and have identified new drug targets. To effectively combat this important pathogen, increased funding is essential. PMID:25769247

  19. 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

  20. 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

  1. 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.

  2. 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

  3. 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.

  4. Nonthermal Biological Treatments Using Discharge Plasma Produced by Pulsed Power 5. Inactivation of Cryptosporidium Oocysts by UV Emission Generated from Pulsed Arc Discharge in Water

    NASA Astrophysics Data System (ADS)

    Kunitomo, Shinta

    Cryptosporidium contaminates most surface waters around the world. It is difficult to remove through conventional treatment processes, and is extremely resistant to the method of chemical disinfection typically used to inactivate these microorganisms. We have developed a new technology for inactivating Cryptosporidium oocysts by using a pulsed arc discharge in water, which creates shock waves, UV emissions, and radicals. The pulsed arc is generated between two cylindrical stainless steel rod electrodes, 6 mm in diameter, and 2 mm apart. We applied this method to the inactivation of oocysts in backwash water from a sand-filter unit of a drinking water plant. The results indicate that the major factor influencing inactivation is UV emissions, and that more than 99% of the oocysts in the high turbidity backwash water (80 NTU) are inactivated with an energy of 0.24 kWh/m3.

  5. 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...

  6. 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...

  7. 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

  8. Cryptosporidium muris in a Texas canine population.

    PubMed

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

    2008-06-01

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

  9. 40 CFR 141.715 - Microbial toolbox options for meeting Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... criteria are in § 141.716(b). Pre Filtration Toolbox Options (3) Presedimentation basin with coagulation 0... separate granular media filtration stage if treatment train includes coagulation prior to first...

  10. 40 CFR 141.715 - Microbial toolbox options for meeting Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... criteria are in § 141.716(b). Pre Filtration Toolbox Options (3) Presedimentation basin with coagulation 0... separate granular media filtration stage if treatment train includes coagulation prior to first...

  11. 40 CFR 141.715 - Microbial toolbox options for meeting Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... criteria are in § 141.716(b). Pre Filtration Toolbox Options (3) Presedimentation basin with coagulation 0... separate granular media filtration stage if treatment train includes coagulation prior to first...

  12. 40 CFR 141.715 - Microbial toolbox options for meeting Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... criteria are in § 141.716(b). Pre Filtration Toolbox Options (3) Presedimentation basin with coagulation 0... separate granular media filtration stage if treatment train includes coagulation prior to first...

  13. 40 CFR 141.715 - Microbial toolbox options for meeting Cryptosporidium treatment requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... criteria are in § 141.716(b). Pre Filtration Toolbox Options (3) Presedimentation basin with coagulation 0... separate granular media filtration stage if treatment train includes coagulation prior to first...

  14. 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

  15. Cryptosporidium (Crypto)

    MedlinePlus

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

  16. 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

  17. 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:25507595

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

    PubMed

    Jex, Aaron R; Gasser, Robin B

    2009-03-01

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

  19. 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

  20. 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.

  1. 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...

  2. 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 ...

  3. 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.

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

    PubMed

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

    2014-02-01

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

  5. 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

  6. 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

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

    PubMed

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

    2007-06-01

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

  8. 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

  9. 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.

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

    PubMed

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

    2015-01-15

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

  11. 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.

  12. Comparative diagnostic techniques for cryptosporidium infection.

    PubMed

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

    2014-02-24

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

  13. 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

  14. 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.

  15. 40 CFR 141.211 - Special notice for repeated failure to conduct monitoring of the source water for Cryptosporidium...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

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

  16. 40 CFR 141.211 - Special notice for repeated failure to conduct monitoring of the source water for Cryptosporidium...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

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

  17. Cryptosporidium species in humans and animals: current understanding and research needs.

    PubMed

    Ryan, Una; Fayer, Ronald; Xiao, Lihua

    2014-11-01

    Cryptosporidium is increasingly recognized as one of the major causes of moderate to severe diarrhoea in developing countries. With treatment options limited, control relies on knowledge of the biology and transmission of the members of the genus responsible for disease. Currently, 26 species are recognized as valid on the basis of morphological, biological and molecular data. Of the nearly 20 Cryptosporidium species and genotypes that have been reported in humans, Cryptosporidium hominis and Cryptosporidium parvum are responsible for the majority of infections. Livestock, particularly cattle, are one of the most important reservoirs of zoonotic infections. Domesticated and wild animals can each be infected with several Cryptosporidium species or genotypes that have only a narrow host range and therefore have no major public health significance. Recent advances in next-generation sequencing techniques will significantly improve our understanding of the taxonomy and transmission of Cryptosporidium species, and the investigation of outbreaks and monitoring of emerging and virulent subtypes. Important research gaps remain including a lack of subtyping tools for many Cryptosporidium species of public and veterinary health importance, and poor understanding of the genetic determinants of host specificity of Cryptosporidium species and impact of climate change on the transmission of Cryptosporidium. PMID:25111501

  18. Molecular characterisation of Cryptosporidium isolates from pet reptiles.

    PubMed

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

    2009-03-23

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

  19. 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...

  20. 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...

  1. 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...

  2. 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...

  3. 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

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

    PubMed

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

    2007-08-01

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

  5. 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...

  6. Cryptosporidium Source Tracking in the Potomac River Watershed▿

    PubMed Central

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

    2008-01-01

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

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

    PubMed

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

    2011-06-30

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

  8. Drug repurposing screen reveals FDA-approved inhibitors of human HMG-CoA reductase and isoprenoid synthesis that block Cryptosporidium parvum growth.

    PubMed

    Bessoff, Kovi; Sateriale, Adam; Lee, K Kyungae; Huston, Christopher D

    2013-04-01

    Cryptosporidiosis, a diarrheal disease usually caused by Cryptosporidium parvum or Cryptosporidium hominis in humans, can result in fulminant diarrhea and death in AIDS patients and chronic infection and stunting in children. Nitazoxanide, the current standard of care, has limited efficacy in children and is no more effective than placebo in patients with advanced AIDS. Unfortunately, the lack of financial incentives and the technical difficulties associated with working with Cryptosporidium parasites have crippled efforts to develop effective treatments. In order to address these obstacles, we developed and validated (Z' score = 0.21 to 0.47) a cell-based high-throughput assay and screened a library of drug repurposing candidates (the NIH Clinical Collections), with the hopes of identifying safe, FDA-approved drugs to treat cryptosporidiosis. Our screen yielded 21 compounds with confirmed activity against C. parvum growth at concentrations of <10 μM, many of which had well-defined mechanisms of action, making them useful tools to study basic biology in addition to being potential therapeutics. Additional work, including structure-activity relationship studies, identified the human 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase inhibitor itavastatin as a potent inhibitor of C. parvum growth (50% inhibitory concentration [IC(50)] = 0.62 μM). Bioinformatic analysis of the Cryptosporidium genomes indicated that the parasites lack all known enzymes required for the synthesis of isoprenoid precursors. Additionally, itavastatin-induced growth inhibition of C. parvum was partially reversed by the addition of exogenous isopentenyl pyrophosphate, suggesting that itavastatin reduces Cryptosporidium growth via on-target inhibition of host HMG-CoA reductase and that the parasite is dependent on the host cell for synthesis of isoprenoid precursors.

  9. Cryptic parasite revealed improved prospects for treatment and control of human cryptosporidiosis through advanced technologies.

    PubMed

    Jex, Aaron R; Smith, Huw V; Nolan, Matthew J; Campbell, Bronwyn E; Young, Neil D; Cantacessi, Cinzia; Gasser, Robin B

    2011-01-01

    Cryptosporidium is an important genus of parasitic protozoa of humans and other vertebrates and is a major cause of intestinal disease globally. Unlike many common causes of infectious enteritis, there are no widely available, effective vaccine or drug-based intervention strategies for Cryptosporidium, and control is focused mainly on prevention. This approach is particularly deficient for infections of severely immunocompromised and/or suppressed, the elderly or malnourished people. However, cryptosporidiosis also presents a significant burden on immunocompetent individuals, and can, for example have lasting effects on the physical and mental development of children infected at an early age. In the last few decades, our understanding of Cryptosporidium has expanded significantly in numerous areas, including the parasite life-cycle, the processes of excystation, cellular invasion and reproduction, and the interplay between parasite and host. Nonetheless, despite extensive research, many aspects of the biology of Cryptosporidium remain unknown, and treatment and control are challenging. Here, we review the current state of knowledge of Cryptosporidium, with a focus on major advances arising from the recently completed genome sequences of the two species of greatest relevance in humans, namely Cryptosporidium hominis and Cryptosporidium parvum. In addition, we discuss the potential of next-generation sequencing technologies, new advances in in silico analyses and progress in in vitro culturing systems to bridge these gaps and to lead toward effective treatment and control of cryptosporidiosis.

  10. Nutrient addition to enhance biological treatment of greywater.

    PubMed

    Jefferson, B; Burgess, J E; Pichon, A; Harkness, J; Judd, S J

    2001-08-01

    This study compares the chemical oxygen demand (COD) removal and respiration rates of a microbial population treating real and synthetic greywaters dosed with nutrient supplements. The nutrient composition of the real and synthetic greywaters was analysed and the dosing regime for nitrogen, phosphorus and a range of trace metals planned accordingly. The doses consisted of eight single additives (macronutrients and trace metals) to the control greywater and six trace metal additions to C: N : P balanced greywater. The COD removal for the control real and synthetic greywater in lab-scale activated sludge systems (0.038 and 0.286 kg COD kg MLSS(-1) d(-1), respectively) confirmed nutrient limitation and the poor degree of greywater treatment. Nutrient dosing increased the COD removal rate and oxygen uptake rate in many cases. The greatest stimulation of microbial activity was observed with zinc additions to C: N: P balanced real greywater (1.291 kg COD kg MLSS(-1) d(-1) over 30 times the control). Inhibitory effects to various extents were rare and limited mainly to the additions of metals to synthetic greywater. The dominance of chemicals effects was observed on addition of some micronutrients; notably iron and aluminium, metals on which many coagulants for use in biotreatment of other wastewaters are based. The data indicate that the impact of understanding microbial processes and the nutrients required for wastewater treatment can only serve to optimise process efficiency for the proposed treatment of greywater.

  11. 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...

  12. 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...

  13. 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...

  14. 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...

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

    PubMed Central

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

    1993-01-01

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

  16. Identification of additional MAP kinases activated upon PAMP treatment

    PubMed Central

    Nitta, Yukino; Ding, Pingtao; Zhang, Yuelin

    2014-01-01

    Mitogen-activated protein (MAP) kinase cascades play important roles in plant immunity. Upon pathogen associated molecular pattern (PAMP) treatment, MPK3, MPK6 and MPK4 are quickly activated by upstream MKKs through phosphorylation. Western blot analysis using α-phospho-p44/42-ERK antibody suggests that additional MPKs with similar size as MPK4 are also activated upon PAMP perception. To identify these MAP kinases, 7 candidate MPKs with similar sizes as MPK4 were selected for further analysis. Transgenic plants expressing these MPKs with a ZZ-3xFLAG double tag of 17 kD were generated and analyzed by western blot. MPK1, MPK11 and MPK13 were found to be phosphorylated upon treatment with flg22. Our study revealed additional MAPKs being activated during PAMP-triggered immunity. PMID:25482788

  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.

  19. 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

  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. 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

  2. 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

  3. Presence of Cryptosporidium spp. and Giardia duodenalis through drinking water.

    PubMed

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

    2008-11-01

    To evaluate the presence of Cryptosporidium spp. and Giardia duodenalis in the influent and final effluent of sixteen drinking water treatment plants located in a hydrographic basin in Galicia (NW Spain) - in which the principal river is recognised as a Site of Community Importance (SCI) - estimate the efficiency of treatment plants in removing these protozoans and determine the species and genotypes of the parasites by means of a molecular assay. All plant samples of influent and final effluent (50-100 l) were examined in the spring, summer, autumn and winter of 2007. A total of 128 samples were analysed by method 1623, developed by US Environmental Protection Agency for isolation and detection of both parasites. To identify the genotypes present the following genes were amplified and sequenced: 18S SSU rRNA (Cryptosporidium spp.) and b-giardina (G. duodenalis). The mean concentrations of parasites in the influent were 0.0-10.5 Cryptosporidium spp. oocysts per litre and 1.0-12.8 of G. duodenalis cysts per litre. In the final treated effluent, the mean concentration of parasites ranged from 0.0-3.0 oocysts per litre and 0.5-4.0 cysts per litre. The distribution of results by season revealed that in all plants, the highest numbers of (oo)cysts were recorded in spring and summer. Cryptosporidium parvum, C. andersoni, C. hominis and assemblages A-I, A-II, E of G. duodenalis were detected. Cryptosporidium spp. and G. duodenalis were consistently found at high concentrations in drinking water destined for human and animal consumption in the hydrographic basin under study, in Galicia (NW Spain). It is important that drinking water treatment authorities rethink the relevance of contamination levels of both parasites in drinking water and develop adequate countermeasures.

  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. 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.

  6. 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.

  7. Cryptosporidium infection in solid organ transplantation

    PubMed Central

    Florescu, Diana F; Sandkovsky, Uriel

    2016-01-01

    Diarrhea is a common complication in solid organ transplant (SOT) recipients and may be attributed to immunosuppressive drugs or infectious organisms such as bacteria, viruses or parasites. Cryptosporidium usually causes self-limited diarrhea in immunocompetent hosts. Although it is estimated that cryptosporidium is involved in about 12% of cases of infectious diarrhea in developing countries and causes approximately 748000 cases each year in the United States, it is still an under recognized and important cause of infectious diarrhea in SOT recipients. It may run a protracted course with severe diarrhea, fluid and electrolyte depletion and potential for organ failure. Although diagnostic methodologies have improved significantly, allowing for fast and accurate identification of the parasite, treatment of the disease is difficult because antiparasitic drugs have modest activity at best. Current management includes fluid and electrolyte replacement, reduction of immunosuppression and single therapy with Nitazoxanide or combination therapy with Nitazoxanide and other drugs. Future drug and vaccine development may add to the currently poor armamentarium to manage the disease. The current review highlights key epidemiological, diagnostic and management issues in the SOT population.

  8. Cryptosporidium infection in solid organ transplantation

    PubMed Central

    Florescu, Diana F; Sandkovsky, Uriel

    2016-01-01

    Diarrhea is a common complication in solid organ transplant (SOT) recipients and may be attributed to immunosuppressive drugs or infectious organisms such as bacteria, viruses or parasites. Cryptosporidium usually causes self-limited diarrhea in immunocompetent hosts. Although it is estimated that cryptosporidium is involved in about 12% of cases of infectious diarrhea in developing countries and causes approximately 748000 cases each year in the United States, it is still an under recognized and important cause of infectious diarrhea in SOT recipients. It may run a protracted course with severe diarrhea, fluid and electrolyte depletion and potential for organ failure. Although diagnostic methodologies have improved significantly, allowing for fast and accurate identification of the parasite, treatment of the disease is difficult because antiparasitic drugs have modest activity at best. Current management includes fluid and electrolyte replacement, reduction of immunosuppression and single therapy with Nitazoxanide or combination therapy with Nitazoxanide and other drugs. Future drug and vaccine development may add to the currently poor armamentarium to manage the disease. The current review highlights key epidemiological, diagnostic and management issues in the SOT population. PMID:27683627

  9. Cryptosporidium infection in solid organ transplantation.

    PubMed

    Florescu, Diana F; Sandkovsky, Uriel

    2016-09-24

    Diarrhea is a common complication in solid organ transplant (SOT) recipients and may be attributed to immunosuppressive drugs or infectious organisms such as bacteria, viruses or parasites. Cryptosporidium usually causes self-limited diarrhea in immunocompetent hosts. Although it is estimated that cryptosporidium is involved in about 12% of cases of infectious diarrhea in developing countries and causes approximately 748000 cases each year in the United States, it is still an under recognized and important cause of infectious diarrhea in SOT recipients. It may run a protracted course with severe diarrhea, fluid and electrolyte depletion and potential for organ failure. Although diagnostic methodologies have improved significantly, allowing for fast and accurate identification of the parasite, treatment of the disease is difficult because antiparasitic drugs have modest activity at best. Current management includes fluid and electrolyte replacement, reduction of immunosuppression and single therapy with Nitazoxanide or combination therapy with Nitazoxanide and other drugs. Future drug and vaccine development may add to the currently poor armamentarium to manage the disease. The current review highlights key epidemiological, diagnostic and management issues in the SOT population. PMID:27683627

  10. RENAL AND CLOACAL CRYPTOSPORIDIOSIS (CRYPTOSPORIDIUM AVIAN GENOTYPE V) IN A MAJOR MITCHELL'S COCKATOO (LOPHOCHROA LEADBEATERI).

    PubMed

    Curtiss, Jeffrey B; Leone, Angelique M; Wellehan, James F X; Emerson, Jessica A; Howerth, Elizabeth W; Farina, Lisa L

    2015-12-01

    A 7-yr-old male Major Mitchell's cockatoo (Lophochroa leadbeateri) presented with a recent history of lethargy and anorexia. Physical examination revealed poor body condition and cloacal prolapse. Abnormalities on serum chemistry included severe hyperuricemia and hyperphosphatemia with a low calcium-to-phosphorus ratio. Symptomatic treatment was initiated including intravenous fluids and antibiotics. The bird continued to decline and died within a few days. Visceral gout and renal and cloacal pathology were observed on gross necropsy. Histopathology revealed chronic inflammation within the kidney, ureter, and cloaca in association with protozoal organisms and an invasive cloacal adenocarcinoma tumor. The location and morphology was consistent with Cryptosporidium sp., confirmed by immunohistochemistry and molecular testing. Direct sequencing identified Cryptosporidium avian genotype V. To the author's knowledge, this is the first reported infection of Cryptosporidium avian genotype V associated with clinical disease in birds and the first renal Cryptosporidium infection in a psittacine.

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

    PubMed

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

    2013-05-01

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

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

    PubMed

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

    2013-07-01

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

  13. 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

  14. Potential molecular tools for assessing the public health risk associated with waterborne Cryptosporidium oocysts.

    PubMed

    Kothavade, Rajendra J

    2012-08-01

    The use of multiple barrier stages at water and wastewater treatment facilities allows for the effective removal of the vast majority of coliforms and other enteric and non-enteric microbes. Subsequent disinfection steps (chlorine, ozone and UV irradiation) are utilized to inactivate microbes that escape the preceding treatment stages. Most viruses, bacteria and protozoa, such as Giardia, are effectively inactivated by chlorination; however, Cryptosporidium is relatively more resistant to environmental conditions and to chlorination. Therefore, UV disinfection has been introduced at many water and wastewater treatment plants to increase log inactivation. Any accidental treatment failure may pose a significant risk to public health. Waterborne transmission of coccidian parasites such as Cryptosporidium and Giardia continues to be a major public health concern. No effective therapies currently exist to treat cryptosporidiosis and the global increase in immunocompromised populations has emphasized the need for water utilities and public health laboratories to have immediate and reliable access to highly sensitive test methods that can determine the host specificity, viability and infectivity of protozoa in the water supply. The most common method used for monitoring Cryptosporidium oocysts and Giardia cysts at intermediate treatment stages and in finished drinking water is the US EPA Method 1623. Although Cryptosporidium species are morphologically indistinguishable, they differ greatly in their host specificity and infectivity. Method 1623 provides quantitative information about Cryptosporidium and Giardia contamination but cannot distinguish between species for intervention purposes in outbreak situations, nor is this method reliable for determining whether the oocyst on the slide is infective for humans. Molecular methods have proven valuable in diagnosing infectious diseases, especially those for which the causative agent is difficult to grow in culture, and

  15. 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...

  16. 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...

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

    PubMed

    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.

  18. 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

  19. 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...

  20. Cryptosporidium hominis gene catalog: a resource for the selection of novel Cryptosporidium vaccine candidates

    PubMed Central

    Ifeonu, Olukemi O.; Simon, Raphael; Tennant, Sharon M.; Sheoran, Abhineet S.; Daly, Maria C.; Felix, Victor; Kissinger, Jessica C.; Widmer, Giovanni; Levine, Myron M.; Tzipori, Saul; Silva, Joana C.

    2016-01-01

    Human cryptosporidiosis, caused primarily by Cryptosporidium hominis and a subset of Cryptosporidium parvum, is a major cause of moderate-to-severe diarrhea in children under 5 years of age in developing countries and can lead to nutritional stunting and death. Cryptosporidiosis is particularly severe and potentially lethal in immunocompromised hosts. Biological and technical challenges have impeded traditional vaccinology approaches to identify novel targets for the development of vaccines against C. hominis, the predominant species associated with human disease. We deemed that the existence of genomic resources for multiple species in the genus, including a much-improved genome assembly and annotation for C. hominis, makes a reverse vaccinology approach feasible. To this end, we sought to generate a searchable online resource, termed C. hominis gene catalog, which registers all C. hominis genes and their properties relevant for the identification and prioritization of candidate vaccine antigens, including physical attributes, properties related to antigenic potential and expression data. Using bioinformatic approaches, we identified ∼400 C. hominis genes containing properties typical of surface-exposed antigens, such as predicted glycosylphosphatidylinositol (GPI)-anchor motifs, multiple transmembrane motifs and/or signal peptides targeting the encoded protein to the secretory pathway. This set can be narrowed further, e.g. by focusing on potential GPI-anchored proteins lacking homologs in the human genome, but with homologs in the other Cryptosporidium species for which genomic data are available, and with low amino acid polymorphism. Additional selection criteria related to recombinant expression and purification include minimizing predicted post-translation modifications and potential disulfide bonds. Forty proteins satisfying these criteria were selected from 3745 proteins in the updated C. hominis annotation. The immunogenic potential of a few of these is

  1. 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

  2. Prospects for immunotherapy and vaccines against Cryptosporidium.

    PubMed

    Mead, Jan R

    2014-01-01

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

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

    PubMed

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

    2014-05-20

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

  4. 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

  5. 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

  6. 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

  7. 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

  8. 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.

  9. First molecular characterization of Cryptosporidium in Yemen.

    PubMed

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

    2013-05-01

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

  10. 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.

  11. 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

  12. 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.

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

    DOE PAGES

    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

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

    PubMed

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

    2015-05-01

    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. PMID:25945705

  15. 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...

  16. 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...

  17. 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...

  18. Molecular identification of Cryptosporidium parvum and Giardia duodenalis in the Italian water buffalo (Bubalus bubalis).

    PubMed

    Cacciò, Simone M; Rinaldi, Laura; Cringoli, Giuseppe; Condoleo, Renato; Pozio, Edoardo

    2007-11-30

    Livestock are commonly infected with protozoan parasites of the genera Cryptosporidium and Giardia, and some of the species and genotypes found in these animals have zoonotic significance. We characterized isolates of both parasites recovered from the Italian water buffalo (Bubalus bubalis), an economically important species whose milk is used for the production of "buffalo mozzarella" fresh cheese. Molecular analysis of the Cryptosporidium small subunit ribosomal DNA gene and of the Giardia beta-giardin gene shows the presence of both zoonotic parasites (Cryptosporidium parvum and Giardia duodenalis assemblage A) and host-specific parasites (G. duodenalis assemblage E), suggesting that water buffaloes can contribute to environmental contamination with oocysts and cysts potentially infectious to humans if their faeces are improperly disposed of. On the other hand, mozzarella cheese is probably a safe product, given that its production involves the treatment of cheese curd at 85-95 degrees C, which is likely to kill or inactivate the parasites.

  19. 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.

  20. A year-long study of Cryptosporidium species and subtypes in recreational, drinking and wastewater from the central area of Spain.

    PubMed

    Galván, A L; Magnet, A; Izquierdo, F; Fernández Vadillo, C; Peralta, R H; Angulo, S; Fenoy, S; del Aguila, C

    2014-01-15

    A year-long longitudinal study was undertaken to evaluate the presence of Cryptosporidium spp. in drinking water treatment plants (DWTPs), wastewater treatment plants (WWTPs) and freshwater bathing beaches (FBBs) from the central area of Spain. Water samples were collected according to USEPA Method 1623, and concentrated by the IDEXX Filta-Max® system. Cryptosporidium species were detected based on PCR-restriction fragment length polymorphism and sequence analyses of the ssuRNA gene. C. hominis and/or C. parvum isolates were subtyped by DNA sequencing of the Gp60 gene. Among 150 samples, 23 (15.3%) were positive by IFAT and 40 (26.7%) by PCR. Cryptosporidium spp. was more frequent in WWTPs (26.2 and 50.8%) and FBBs (12.5 and 17.5%) by IFAT and PCR respectively. Effluent waters from DWTPs were negative for this parasite suggesting that they are suitable for public use. Tertiary treatment in the WWTPs demonstrated a high removal efficiency of Cryptosporidium in the samples evaluated. Cryptosporidium species identified included C. hominis, C. parvum, C. ubiquitum, C. andersoni and C. muris. Subtyping analysis revealed C. hominis IbA10G2 and IeA11G3T3 alleles, which is the first report of the latter in water samples. Cryptosporidium highest frequency was observed in winter and spring. Our data provide information about the occurrence and diversity of Cryptosporidium in water of human use from the central area of Spain.

  1. 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...

  2. 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...

  3. Prevalence and genetic characterization of Cryptosporidium isolates from common brushtail possums (Trichosurus vulpecula) adapted to urban settings.

    PubMed

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

    2008-09-01

    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.

  4. Cryptosporidium: Genomic and Biochemical Features

    PubMed Central

    Rider, Stanley Dean; Zhu, Guan

    2010-01-01

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

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

    PubMed

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

    2006-06-01

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

  6. 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

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

    PubMed

    Reina, Fábio Tadeu Rodrigues; Ribeiro, Camila Aparecida; Araújo, Ronalda Silva de; 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

  8. 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...

  9. 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...

  10. New cryptosporidium genotypes in HIV-infected persons.

    PubMed Central

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

    1999-01-01

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

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

    PubMed Central

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

    2005-01-01

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

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

    PubMed

    Edwinson, Adam; Widmer, Giovanni; McEvoy, John

    2016-01-01

    The apicomplexan parasite Cryptosporidium causes cryptosporidiosis, a diarrheal disease that can become chronic and life threatening in immunocompromised and malnourished people. There is no effective drug treatment for those most at risk of severe cryptosporidiosis. The disease pathology is due to a repeated cycle of host cell invasion and parasite replication that amplifies parasite numbers and destroys the intestinal epithelium. This study aimed to better understand the Cryptosporidium replication cycle by identifying molecules that trigger the switch from invasive sporozoite to replicative trophozoite. Our approach was to treat sporozoites of Cryptosporidium parvum and Cryptosporidium hominis, the species causing most human cryptosporidiosis, with various media under axenic conditions and examine the parasites for rounding and nuclear division as markers of trophozoite development and replication, respectively. FBS had a concentration-dependent effect on trophozoite development in both species. Trophozoite development in C. parvum, but not C. hominis, was enhanced when RPMI supplemented with 10% FBS (RPMI-FBS) was conditioned by HCT-8 cells for 3h. The effect of non-conditioned and HCT-8 conditioned RPMI-FBS on trophozoite development was abrogated by proteinase K and sodium metaperiodate pretreatment, indicating a glycoprotein trigger. Cryptosporidium parvum and C. hominis trophozoite development also was triggered by Gal-GalNAc in a concentration-dependent manner. Cryptosporidium parvum replication was greatest following treatments with Gal-GalNAc, followed by conditioned RPMI-FBS and non-conditioned RPMI-FBS (P<0.05). Cryptosporidium hominis replication was significantly less than that in C. parvum for all treatments (P<0.05), and was greatest at the highest tested concentration of Gal-GalNAc (1mM).

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

    PubMed

    Edwinson, Adam; Widmer, Giovanni; McEvoy, John

    2016-01-01

    The apicomplexan parasite Cryptosporidium causes cryptosporidiosis, a diarrheal disease that can become chronic and life threatening in immunocompromised and malnourished people. There is no effective drug treatment for those most at risk of severe cryptosporidiosis. The disease pathology is due to a repeated cycle of host cell invasion and parasite replication that amplifies parasite numbers and destroys the intestinal epithelium. This study aimed to better understand the Cryptosporidium replication cycle by identifying molecules that trigger the switch from invasive sporozoite to replicative trophozoite. Our approach was to treat sporozoites of Cryptosporidium parvum and Cryptosporidium hominis, the species causing most human cryptosporidiosis, with various media under axenic conditions and examine the parasites for rounding and nuclear division as markers of trophozoite development and replication, respectively. FBS had a concentration-dependent effect on trophozoite development in both species. Trophozoite development in C. parvum, but not C. hominis, was enhanced when RPMI supplemented with 10% FBS (RPMI-FBS) was conditioned by HCT-8 cells for 3h. The effect of non-conditioned and HCT-8 conditioned RPMI-FBS on trophozoite development was abrogated by proteinase K and sodium metaperiodate pretreatment, indicating a glycoprotein trigger. Cryptosporidium parvum and C. hominis trophozoite development also was triggered by Gal-GalNAc in a concentration-dependent manner. Cryptosporidium parvum replication was greatest following treatments with Gal-GalNAc, followed by conditioned RPMI-FBS and non-conditioned RPMI-FBS (P<0.05). Cryptosporidium hominis replication was significantly less than that in C. parvum for all treatments (P<0.05), and was greatest at the highest tested concentration of Gal-GalNAc (1mM). PMID:26432292

  14. Cryptosporidium in humans and animals-a one health approach to prophylaxis.

    PubMed

    Ryan, U; Zahedi, A; Paparini, A

    2016-09-01

    Cryptosporidium is a major cause of moderate-to-severe diarrhoea in humans worldwide, second only to rotavirus. Due to the wide host range and environmental persistence of this parasite, cryptosporidiosis can be zoonotic and associated with foodborne and waterborne outbreaks. Currently, 31 species are recognized as valid, and of these, Cryptosporidium hominis and Cryptosporidium parvum are responsible for the majority of infections in humans. The immune status of the host, both innate and adaptive immunity, has a major impact on the severity of the disease and its prognosis. Immunocompetent individuals typically experience self-limiting diarrhoea and transient gastroenteritis lasting up to 2 weeks and recover without treatment, suggesting an efficient host antiparasite immune response. Immunocompromised individuals can suffer from intractable diarrhoea, which can be fatal. Effective drug treatments and vaccines are not yet available. As a result of this, the close cooperation and interaction between veterinarians, health physicians, environmental managers and public health operators is essential to properly control this disease. This review focuses on a One Health approach to prophylaxis, including the importance of understanding transmission routes for zoonotic Cryptosporidium species, improved sanitation and better risk management, improved detection, diagnosis and treatment and the prospect of an effective anticryptosporidial vaccine. PMID:27454991

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

    PubMed

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

    2012-07-01

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

  16. 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

  17. 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.

  18. 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...

  19. 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

  20. Cryptosporidium and Toxoplasma Parasites Are Inhibited by a Benzoxaborole Targeting Leucyl-tRNA Synthetase

    PubMed Central

    Liu, Ru-Juan; Lukarska, Maria; Gut, Jiri; Bougdour, Alexandre; Touquet, Bastien; Wang, En-Duo; Li, Xianfeng; Alley, M. R. K.; Freund, Yvonne R.; Rosenthal, Philip J.; Hakimi, Mohamed-Ali

    2016-01-01

    The apicomplexan parasites Cryptosporidium and Toxoplasma are serious threats to human health. Cryptosporidiosis is a severe diarrheal disease in malnourished children and immunocompromised individuals, with the only FDA-approved drug treatment currently being nitazoxanide. The existing therapies for toxoplasmosis, an important pathology in immunocompromised individuals and pregnant women, also have serious limitations. With the aim of developing alternative therapeutic options to address these health problems, we tested a number of benzoxaboroles, boron-containing compounds shown to be active against various infectious agents, for inhibition of the growth of Cryptosporidium parasites in mammalian cells. A 3-aminomethyl benzoxaborole, AN6426, with activity in the micromolar range and with activity comparable to that of nitazoxanide, was identified and further characterized using biophysical measurements of affinity and crystal structures of complexes with the editing domain of Cryptosporidium leucyl-tRNA synthetase (LeuRS). The same compound was shown to be active against Toxoplasma parasites, with the activity being enhanced in the presence of norvaline, an amino acid that can be mischarged by LeuRS. Our observations are consistent with AN6426 inhibiting protein synthesis in both Cryptosporidium and Toxoplasma by forming a covalent adduct with tRNALeu in the LeuRS editing active site and suggest that further exploitation of the benzoxaborole scaffold is a valid strategy to develop novel, much needed antiparasitic agents. PMID:27431220

  1. Cryptosporidium and Toxoplasma Parasites Are Inhibited by a Benzoxaborole Targeting Leucyl-tRNA Synthetase.

    PubMed

    Palencia, Andrés; Liu, Ru-Juan; Lukarska, Maria; Gut, Jiri; Bougdour, Alexandre; Touquet, Bastien; Wang, En-Duo; Li, Xianfeng; Alley, M R K; Freund, Yvonne R; Rosenthal, Philip J; Hakimi, Mohamed-Ali; Cusack, Stephen

    2016-10-01

    The apicomplexan parasites Cryptosporidium and Toxoplasma are serious threats to human health. Cryptosporidiosis is a severe diarrheal disease in malnourished children and immunocompromised individuals, with the only FDA-approved drug treatment currently being nitazoxanide. The existing therapies for toxoplasmosis, an important pathology in immunocompromised individuals and pregnant women, also have serious limitations. With the aim of developing alternative therapeutic options to address these health problems, we tested a number of benzoxaboroles, boron-containing compounds shown to be active against various infectious agents, for inhibition of the growth of Cryptosporidium parasites in mammalian cells. A 3-aminomethyl benzoxaborole, AN6426, with activity in the micromolar range and with activity comparable to that of nitazoxanide, was identified and further characterized using biophysical measurements of affinity and crystal structures of complexes with the editing domain of Cryptosporidium leucyl-tRNA synthetase (LeuRS). The same compound was shown to be active against Toxoplasma parasites, with the activity being enhanced in the presence of norvaline, an amino acid that can be mischarged by LeuRS. Our observations are consistent with AN6426 inhibiting protein synthesis in both Cryptosporidium and Toxoplasma by forming a covalent adduct with tRNA(Leu) in the LeuRS editing active site and suggest that further exploitation of the benzoxaborole scaffold is a valid strategy to develop novel, much needed antiparasitic agents.

  2. 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.

  3. 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

  4. Cryptosporidium and Toxoplasma Parasites Are Inhibited by a Benzoxaborole Targeting Leucyl-tRNA Synthetase.

    PubMed

    Palencia, Andrés; Liu, Ru-Juan; Lukarska, Maria; Gut, Jiri; Bougdour, Alexandre; Touquet, Bastien; Wang, En-Duo; Li, Xianfeng; Alley, M R K; Freund, Yvonne R; Rosenthal, Philip J; Hakimi, Mohamed-Ali; Cusack, Stephen

    2016-10-01

    The apicomplexan parasites Cryptosporidium and Toxoplasma are serious threats to human health. Cryptosporidiosis is a severe diarrheal disease in malnourished children and immunocompromised individuals, with the only FDA-approved drug treatment currently being nitazoxanide. The existing therapies for toxoplasmosis, an important pathology in immunocompromised individuals and pregnant women, also have serious limitations. With the aim of developing alternative therapeutic options to address these health problems, we tested a number of benzoxaboroles, boron-containing compounds shown to be active against various infectious agents, for inhibition of the growth of Cryptosporidium parasites in mammalian cells. A 3-aminomethyl benzoxaborole, AN6426, with activity in the micromolar range and with activity comparable to that of nitazoxanide, was identified and further characterized using biophysical measurements of affinity and crystal structures of complexes with the editing domain of Cryptosporidium leucyl-tRNA synthetase (LeuRS). The same compound was shown to be active against Toxoplasma parasites, with the activity being enhanced in the presence of norvaline, an amino acid that can be mischarged by LeuRS. Our observations are consistent with AN6426 inhibiting protein synthesis in both Cryptosporidium and Toxoplasma by forming a covalent adduct with tRNA(Leu) in the LeuRS editing active site and suggest that further exploitation of the benzoxaborole scaffold is a valid strategy to develop novel, much needed antiparasitic agents. PMID:27431220

  5. Target validation of the inosine monophosphate dehydrogenase (IMPDH) gene in Cryptosporidium using Phylomer(®) peptides.

    PubMed

    Jefferies, R; Yang, R; Woh, C K; Weldt, T; Milech, N; Estcourt, A; Armstrong, T; Hopkins, R; Watt, P; Reid, S; Armson, A; Ryan, U M

    2015-01-01

    Cryptosporidiosis, a gastroenteric disease characterised mainly by diarrheal illnesses in humans and mammals is caused by infection with the protozoan parasite Cryptosporidium. Treatment options for cryptosporidiosis are limited, with the current therapeutic nitazoxanide, only partly efficacious in immunocompetent individuals. The parasite lacks de novo purine synthesis, and is exclusively dependant on purine salvage from its host. Inhibition of the inosine 5' monophosphate dehydrogenase (IMPDH), a purine salvage enzyme that is essential for DNA synthesis, thereby offers a potential drug target against this parasite. In the present study, a yeast-two-hybrid system was used to identify Phylomer peptides within a library constructed from the genomes of 25 phylogenetically diverse bacteria that targeted the IMPDH of Cryptosporidium parvum (IMPcp) and Cryptosporidium hominis (IMPch). We identified 38 unique interacting Phylomers, of which, 12 were synthesised and screened against C. parvum in vitro. Two Phylomers exhibited significant growth inhibition (81.2-83.8% inhibition; P < 0.05), one of which consistently exhibited positive interactions with IMPcp and IMPch during primary and recapitulation yeast two-hybrid screening and did not interact with either of the human IMPDH proteins. The present study highlightsthe potential of Phylomer peptides as target validation tools for Cryptosporidium and other organisms and diseases because of their ability to bind with high affinity to target proteins and disrupt function. PMID:25447124

  6. 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.

  7. 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

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

    PubMed

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

    2014-03-01

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

  9. 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.

  10. Identification of Cryptosporidium oocysts in river water.

    PubMed Central

    Ongerth, J E; Stibbs, H H

    1987-01-01

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

  11. 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...

  12. 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, ...

  13. 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...

  14. 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 ...

  15. 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.

  16. 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

  17. Cryptosporidium species in Australian wildlife and domestic animals.

    PubMed

    Ryan, Una; Power, Michelle

    2012-11-01

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

  18. 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

  19. 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

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

    PubMed

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

    2009-12-23

    Eimeria and Cryptosporidium are among the most common bovine parasites in the world, but little is known about them in Estonia. Basic field research is needed to gain insight into pathogen dynamics, providing knowledge for veterinarians and research. A survey of 45 Estonian dairy farms in 15 counties was carried out between 2006 and 2007. Three age groups: <3, 3-12, and >12 months old animals were sampled. Collected faeces were examined by quantitative flotation and Ziehl-Neelsen contrast staining, and species examined morphologically. Selected samples containing Cryptosporidium were additionally examined by polymerase-chain-reaction (PCR) and sequencing to determine genotypes. Twelve species of Eimeria were identified, seven previously unknown in Estonia. Main species in samples were E. bovis (30%), E. zuernii (23%), and E. ellipsoidalis (14%). All herds were infected and animals aged 3-12 months were more commonly infected with Eimeria oocysts (63%) than any other group. Calves <3 months shed most oocyst, but high counts were rare. A negative association (slope=-0.16, p<0.001) was found between the number of animals infected with Eimeria and the age category. Cryptosporidium were detected in 84% of the farms, and C. andersoni and C. parvum were successfully identified. Though prevalences of Cryptosporidium in the age groups were similar to the sample prevalence (30%) an increase in the infections was found with increasing age (p<0.001). Higher diarrhoea scores were negatively associated with Eimeria spp. infection (slope=-0.08, p<0.05), whereas Cryptosporidium could not be associated with the presence of diarrhoea. Frequent low intensity infections of animals in all age groups with both parasites apply a constant stress on the animals with impact on health and production. The Estonian results are different compared to available studies in regard of: older animals commonly being infected, finding of modest oocyst counts, and distribution of Eimeria species.

  1. 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

  2. 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

  3. Cryptosporidiosis in Kuwaiti children: association of clinical characteristics with Cryptosporidium species and subtypes.

    PubMed

    Iqbal, Jamshaid; Khalid, Nabila; Hira, Parsotam Ravjee

    2011-05-01

    To determine the association of clinical characteristics with Cryptosporidium types and subtypes, faecal specimens from 2548 children with diarrhoea were screened by microscopy for Cryptosporidium spp., and positive specimens were genotyped and subtyped by PCR-RFLP. A total of 87 of the 2548 children (3.4 %) had cryptosporidial diarrhoea by microscopy and the majority (41.4 %) of the infected children were in the 4-8-year-old age group. Molecular characterization of the 83 children studied further (4 had mixed infections and were not subtyped) showed that Cryptosporidium parvum was the most commonly identified species (73.5 %) and consisted of three subtypes: IIa and IId were the most common (80.3 %), followed by IIc. Twenty-two (26.5 %) of the children had Cryptosporidium hominis and showed three subtypes: Id was the most common (54.5 %), followed by Ia (36.4 %) and Ie. Associated clinical manifestations varied between C. parvum and C. hominis. Diarrhoea associated with subtype Id, the most commonly identified C. hominis subtype, was more severe than that associated with other subtypes. In conclusion, this study confirmed a very different Cryptosporidium genotype and subtype distribution compared with other tropical countries among Kuwaiti children with diarrhoea, with a predominance of C. parvum IIa and IId. In addition, subtype Id of C. hominis was associated with more diverse and severe clinical manifestations in infected children, suggesting that parasite genetics may play an important role in the clinical manifestations of human cryptosporidiosis.

  4. 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

  5. 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.

  6. 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.

  7. 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

  8. 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.

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

    PubMed

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

    2016-10-01

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

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

    PubMed

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

    2016-10-01

    Cryptosporidium is a genus of enteric protozoan parasites of medical and veterinary importance, whose oocysts have been reported to occur in different types of water worldwide, offering a great resistant to the water treatment processes. Heterogeneous solar photocatalysis using titanium dioxide (TiO2) slurry was evaluated on inactivation of Cryptosporidium parvum oocysts in water. Suspensions of TiO2 (0, 63, 100 and 200mg/L) in distilled water (DW) or simulated municipal wastewater treatment plant (MWTP) effluent spiked with C. parvum oocysts were exposed to simulated solar radiation. The use of TiO2 slurry at concentrations of 100 and 200mg/L in DW yielded a high level of oocyst inactivation after 5h of exposure (4.16±2.35% and 15.03±4.54%, respectively, vs 99.33±0.58%, initial value), representing a good improvement relative to the results obtained in the samples exposed without TiO2 (51.06±9.35%). However, in the assays carried out using simulated MWTP effluent, addition of the photocatalyst did not offer better results. Examination of the samples under bright field and epifluorescence microscopy revealed the existence of aggregates comprising TiO2 particles and parasitic forms, which size increased as the concentration of catalyst and the exposure time increased, while the intensity of fluorescence of the oocyst walls decreased. After photocatalytic disinfection process, the recovery of TiO2 slurry by sedimentation provided a substantial reduction in the parasitic load in treated water samples (57.81±1.10% and 82.10±2.64% for 200mg/L of TiO2 in DW and in simulated MWTP effluent, respectively). Although further studies are need to optimize TiO2 photocatalytic disinfection against Cryptosporidium, the results obtained in the present study show the effectiveness of solar photocatalysis using TiO2 slurry in the inactivation of C. parvum oocysts in distilled water. PMID:27543761

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

    PubMed

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

    2000-01-01

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

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

    PubMed

    Ongerth, Jerry E

    2013-09-17

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

  13. 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

  14. Common occurrence of a unique Cryptosporidium ryanae variant in zebu cattle and water buffaloes in the buffer zone of the Chitwan National Park, Nepal.

    PubMed

    Feng, Yaoyu; Karna, Sandeep Raj; Dearen, Theresa K; Singh, Dinesh Kumar; Adhikari, Lekh Nath; Shrestha, Aruna; Xiao, Lihua

    2012-04-30

    There are very few studies on the diversity and public health significance of Cryptosporidium species in zebu cattle and water buffaloes in developing countries. In this study, PCR-restriction fragment length polymorphism and DNA sequence analyses of the small-subunit (SSU) rRNA gene were used to genotype Cryptosporidium specimens from 12 zebu cattle calves, 16 water buffalo calves, and four swamp deer (Cervus duvaucelii) collected from the buffer zone of the Chitwan National Park, Nepal. All Cryptosporidium specimens from cattle and buffaloes belonged to Cryptosporidium ryanae, whereas those from deer belonged to Cryptosporidium ubiquitum. Comparison of the SSU rRNA gene sequences obtained with those from earlier studies has identified a nucleotide substitution unique to all C. ryanae isolates from Nepal, in addition to some sequence heterogeneity among different copies of the gene. The finding of the dominance of a unique C. ryanae variant in both zebu cattle and water buffaloes in Nepal indicates that there is unique cryptosporidiosis transmission in bovine animals in the study area, and cross-species transmission of some Cryptosporidium spp. can occur between related animal species sharing the same habitats.

  15. Cryptosporidium Parvum Transport Through Natural Porous Media

    NASA Astrophysics Data System (ADS)

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

    2005-12-01

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

  16. Treatment of dentoskeletal bimaxillary protrusion: additional extraction of maxillary second molars.

    PubMed

    Iijima, Shigeki; Yoshida, Mitsuru; Terada, Kazuto

    2009-07-01

    This case report describes the orthodontic treatment of an 18-year-old female patient with Class II malocclusion with dentoskeletal bimaxillary protrusion. An acceptable treatment result was obtained with a four first premolar extraction and an additional maxillary two second molar extraction plan. Standard edgewise appliances were placed after the premolars were extracted. A positive soft tissue response to treatment was achieved, and the patient's profile was improved, with a reduction of lip protrusion and mentalis strain. Dentally, the interincisal angle improved significantly as both the maxillary and mandibular incisors were uprighted after space closure. The occlusion remained stable during 2 years of follow-up.

  17. Requirement for additional treatment for dogs with atopic dermatitis undergoing allergen-specific immunotherapy.

    PubMed

    Colombo, S; Hill, P B; Shaw, D J; Thoday, K L

    2007-06-23

    Allergen-specific immunotherapy (ASIT) is one of the main treatments for atopic dermatitis in dogs, but it often requires additional treatments such as antibacterial and antifungal therapy for secondary bacterial and yeast infections, or antipruritic drugs to control the clinical signs or treat the adverse effects of the immunotherapy. Twenty-seven dogs enrolled in a study of ASIT were clinically assessed four times over a period of nine months; their requirement for treatment for secondary bacterial and yeast infections, for the administration of glucocorticoids as additional antipruritic therapy, and for the treatment of any adverse effects of the ASIT were evaluated. Twenty (74 per cent) of the dogs were treated for superficial bacterial pyoderma, 18 (66.6 per cent) required treatment for Malassezia species dermatitis on one or more occasions, eight (29.6 per cent) required treatment for otitis externa due to Malassezia species or bacteria, and eight required glucocorticoids to control their clinical signs. Five (18.5 per cent) of the dogs experienced adverse effects due to the ASIT and two required treatment with antihistamines (H1 receptor antagonists) in order to continue with the ASIT. PMID:17586789

  18. 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.

  19. Additive Effects of Combination Treatment with Anti-inflammatory and Neuroprotective Agents in Experimental Autoimmune Encephalomyelitis

    PubMed Central

    Du, Sienmi; Sandoval, Francisco; Trinh, Pauline; Voskuhl, Rhonda R.

    2011-01-01

    We studied the effects of combination treatment with an anti-inflammatory agent, interferon(IFN)-β, and a putative neuroprotective agent, an estrogen receptor(ER)-β ligand, during EAE. Combination treatment significantly attenuated EAE disease severity, preserved axonal densities in spinal cord, and reduced CNS inflammation. Combining ERβ treatment with IFNβ reduced IL-17, while it abrogated IFNβ-mediated increases in Th1 and Th2 cytokines from splenocytes. Additionally, combination treatment reduced VLA-4 expression on CD4+ T cells, while it abrogated IFNβ-mediated decreases in MMP-9. Our data demonstrate that combination treatments can result in complex effects that could not have been predicted based on monotherapy data alone. PMID:20006910

  20. Detection of Cryptosporidium parvum in soil extracts.

    PubMed

    Walker, Mark; Redelman, Douglas

    2004-03-01

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

  1. 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

  2. Analysis of Cryptosporidium parvum oocyst transport in porous media

    NASA Astrophysics Data System (ADS)

    Kim, Song-Bae; Yavuz Corapcioglu, M.

    2004-08-01

    Cryptosporidium parvum is a protozoan parasite, transmitted through aqueous environments in the form of an oocyst. In this study, a transport model into which sorption, filtration and inactivation mechanisms are incorporated is applied to simulate laboratory column data, and the suitability of a kinetic model to describe the C. parvum oocyst transport and removal in porous media is compared with an equilibrium model. The kinetic model is applied to simulate previous column experimental data and successfully simulates the concentration peak; the late time tailing effect appeared in the breakthrough curves, indicating that the kinetic model is more suitable than the equilibrium one at simulating the fate and transport of the oocysts in porous media. Simulation illustrates that sorption causes retardation along with a tailing in the breakthrough curve. Additionally, filtration acts as a major mechanism of removing the oocysts from the aqueous phase, whereas the role of inactivation in reducing the viable oocyst concentration is minimal.

  3. 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.

  4. 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...

  5. 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...

  6. The food additive-free diet in the treatment of behavior disorders: a review.

    PubMed

    Wender, E H

    1986-02-01

    A food additive-free diet has been recommended as a component of the treatment of developmental/behavioral disorders in children. This diet was initially developed by Dr. Benjamin Feingold for the treatment of aspirin sensitivity in adults, and then extended by him to the management, particularly, of hyperactivity and learning disability in children. The rationale for this diet and the methods employed in investigating its use are described. The claimed therapeutic effects of this diet have been investigated in a number of well-designed studies reviewed here. These studies generally refute a causal association between food additives and behavioral disturbance in children. Suggestions are made regarding the approach towards food additive-free diet therapy in the management of developmental/behavioral disorders.

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

    PubMed

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

    2008-09-01

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

  8. Short communication: effect of pomegranate-residue supplement on Cryptosporidium parvum oocyst shedding in neonatal calves.

    PubMed

    Weyl-Feinstein, S; Markovics, A; Eitam, H; Orlov, A; Yishay, M; Agmon, R; Miron, J; Izhaki, I; Shabtay, A

    2014-09-01

    Cryptosporidium parvum is considered one of the most common enteropathogens, responsible for the high incidence of diarrhea and deleterious implications on immunity and health in neonatal calves. The pomegranate is well known for its health-promoting properties. Two experiments were designed to test the antiparasitical and antidiarrheal effects of concentrated pomegranate extract (CPE) supplement in milk in neonatal Holstein calves. Forty-one calves were randomly divided into control (n=20) and treatment (n=21) groups. For the first experiment, the treatment group was supplemented with 3.75% CPE in the daily milk ration, between 3 and 14 d of age, whereas the control group received only milk. Fecal samples were collected between d 5 and 13 to quantify Cryptosporidium oocysts, and the duration and intensity of diarrhea were evaluated. Reduced fecal oocyst count and diarrhea intensity and duration were revealed in the 3.75% CPE calves. No difference was noted in average daily gain between groups. In a second experiment, which was designed to test the effect of a lower CPE concentration (0.6% of daily milk allocation), no effects on fecal oocyst count and average daily gain were observed. However, compared with control, the lower CPE group was characterized by a shorter duration of diarrhea and higher weight gain among males at 14 d of age. These results suggest that the CPE supplement-to-calf milk ratio may potentially alleviate intestinal morbidity caused by Cryptosporidium. PMID:24952772

  9. Persistence of Nontuberculous Mycobacteria in a Drinking Water System after Addition of Filtration Treatment

    PubMed Central

    Hilborn, Elizabeth D.; Covert, Terry C.; Yakrus, Mitchell A.; Harris, Stephanie I.; Donnelly, Sandra F.; Rice, Eugene W.; Toney, Sean; Bailey, Stephanie A.; Stelma, Gerard N.

    2006-01-01

    There is evidence that drinking water may be a source of infections with pathogenic nontuberculous mycobacteria (NTM) in humans. One method by which NTM are believed to enter drinking water distribution systems is by their intracellular colonization of protozoa. Our goal was to determine whether we could detect a reduction in the prevalence of NTM recovered from an unfiltered surface drinking water system after the addition of ozonation and filtration treatment and to characterize NTM isolates by using molecular methods. We sampled water from two initially unfiltered surface drinking water treatment plants over a 29-month period. One plant received the addition of filtration and ozonation after 6 months of sampling. Sample sites included those at treatment plant effluents, distributed water, and cold water taps (point-of-use [POU] sites) in public or commercial buildings located within each distribution system. NTM were recovered from 27% of the sites. POU sites yielded the majority of NTM, with >50% recovery despite the addition of ozonation and filtration. Closely related electrophoretic groups of Mycobacterium avium were found to persist at POU sites for up to 26 months. Water collected from POU cold water outlets was persistently colonized with NTM despite the addition of ozonation and filtration to a drinking water system. This suggests that cold water POU outlets need to be considered as a potential source of chronic human exposure to NTM. PMID:16957205

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

    PubMed

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

    2013-12-01

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

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

    PubMed

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

    2016-05-01

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

  12. 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

  13. Cryptosporidium infection causes undernutrition and, conversely, weanling undernutrition intensifies infection.

    PubMed

    Coutinho, Bruna P; Oriá, Reinaldo B; Vieira, Carlos M G; Sevilleja, Jesus Emmanuel A D; Warren, Cirle A; Maciel, Jamilly G; Thompson, Meghan R; Pinkerton, Relana C; Lima, Aldo A M; Guerrant, Richard L

    2008-12-01

    Cryptosporidium parvum is a leading pathogen in children in developing countries. To investigate whether early postnatal malnutrition leads to heavier C. parvum infections, we assessed intestinal adaptation and parasite load in suckling mice during the first 2 wk of life, analogous to the first postnatal yr in humans. Undernutrition was induced by daily C57BL6J pup separation from lactating dams. Half of the pups were separated daily, for 4 hr on day 4, 8 hr on day 5, and for 12 hr from day 6 until day 14. On day 6, each pup received an oral inoculum of 10(5) to 10(7) parasites in 10-25 microl of PBS. Littermate controls received PBS alone. Stools were assessed from days 8, 11, and 14 for oocyst counts. Mice were killed on day 14, 8 days postinoculation, at the peak of the infection. Ileal and colon segments were obtained for histology, real-time and reverse transcriptase PCR, and immunoassays. Villus and crypt lengths and cross-sectional areas were also measured. Undernourished and nourished mice infected with excysted 10(6) or 10(7) oocysts exhibited the poorest growth outcomes compared with their uninfected controls. Nourished 10(6)-infected mice had comparable weight decrements to uninfected undernourished mice. Body weight and villi were additively affected by malnutrition and cryptosporidiosis. Hyperplastic crypts and heavier inflammatory responses were found in the ilea of infected malnourished mice. Undernourished infected mice exhibited greater oocyst shedding, TNF-alpha and IFN-gamma intestinal levels, and mRNA expression compared to nourished mice infected with either 10(5) or 10(6) oocysts. Taken together, these findings show that Cryptosporidium infection can cause undernutrition and, conversely, that weanling undernutrition intensifies infection and mucosal damage.

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

    PubMed Central

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

    2013-01-01

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

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

    PubMed

    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.

  16. 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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

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

    2016-08-01

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

  19. 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

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

    PubMed

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

    2016-08-01

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

  1. 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

  2. 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

  3. Rotifers ingest oocysts of Cryptosporidium parvum

    USGS Publications Warehouse

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

    2000-01-01

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

  4. The impact of bismuth addition to sequential treatment on Helicobacter pylori eradication: A pilot study.

    PubMed

    Basyigit, Sebahat; Kefeli, Ayse; Sapmaz, Ferdane; Yeniova, Abdullah Ozgür; Asilturk, Zeliha; Hokkaomeroglu, Murat; Uzman, Metin; Nazligul, Yasar

    2015-10-25

    The success of the current anti-Helicobacter pylori (H. pylori) treatment protocols is reported to decrease by years, and research is needed to strengthen the H. pylori eradication treatment. Sequential treatment (ST), one of the treatment modalities for H. pylori eradication, includes amoxicillin 1 gr b.i.d and proton pump inhibitor b.i.d for first 5 days and then includes clarithromycin 500 mg b.i.d, metronidazole 500 mg b.i.d and a proton pump inhibitor b.i.d for remaining 5 days. In this study, we investigated efficacy and tolerability of bismuth addition in to ST. We included patients that underwent upper gastrointestinal endoscopy in which H. pylori infection was diagnosed by histological examination of antral and corporal gastric mucosa biopsy. Participants were randomly administered ST or bismuth containing ST (BST) protocols for the first-line H. pylori eradication therapy. Participants have been tested by urea breath test for eradication success 6 weeks after the completion of treatment. One hundred and fifty patients (93 female, 57 male) were enrolled. There were no significant differences in eradication rates for both intention to treat population (70.2%, 95% confidence interval [CI]: 66.3-74.1% vs. 71.8%, 95% CI: 61.8-81.7%, for ST and BST, respectively, p>0.05) and per protocol population (74.6%, 95% CI: 63.2-85.8% vs. 73.7%, 95% CI: 63.9-83.5% for ST and BST, respectively, p>0.05). Despite the undeniable effect of bismuth, there may be several possible reasons of unsatisfactory eradication success. Drug administration time, coadministration of other drugs, possible H. pylori resistance to bismuth may affect the eradication success. The addition of bismuth subcitrate to ST regimen does not provide significant increase in eradication rates.

  5. The first recorded outbreak of cryptosporidiosis due to Cryptosporidium cuniculus (formerly rabbit genotype), following a water quality incident.

    PubMed

    Puleston, Richard L; Mallaghan, Cathy M; Modha, Deborah E; Hunter, Paul R; Nguyen-Van-Tam, Jonathan S; Regan, Christopher M; Nichols, Gordon L; Chalmers, Rachel M

    2014-03-01

    We report the first identified outbreak of cryptosporidiosis with Cryptosporidium cuniculus following a water quality incident in Northamptonshire, UK. A standardised, enhanced Cryptosporidium exposure questionnaire was administered to all cases of cryptosporidiosis after the incident. Stool samples, water testing, microscopy slides and rabbit gut contents positive for Cryptosporidium were typed at the Cryptosporidium Reference Unit, Singleton Hospital, Swansea. Twenty-three people were microbiologically linked to the incident although other evidence suggests an excess of 422 cases of cryptosporidiosis above baseline. Most were adult females; unusually for cryptosporidiosis there were no affected children identified under the age of 5 years. Water consumption was possibly higher than in national drinking water consumption patterns. Diarrhoea duration was negatively correlated to distance from the water treatment works where the contamination occurred. Oocyst counts were highest in water storage facilities. This outbreak is the first caused by C. cuniculus infection to have been noted and it has conclusively demonstrated that this species can be a human pathogen. Although symptomatically similar to cryptosporidiosis from C. parvum or C. hominis, this outbreak has revealed some differences, in particular no children under 5 were identified and females were over-represented. These dissimilarities are unexplained although we postulate possible explanations.

  6. The first recorded outbreak of cryptosporidiosis due to Cryptosporidium cuniculus (formerly rabbit genotype), following a water quality incident.

    PubMed

    Puleston, Richard L; Mallaghan, Cathy M; Modha, Deborah E; Hunter, Paul R; Nguyen-Van-Tam, Jonathan S; Regan, Christopher M; Nichols, Gordon L; Chalmers, Rachel M

    2014-03-01

    We report the first identified outbreak of cryptosporidiosis with Cryptosporidium cuniculus following a water quality incident in Northamptonshire, UK. A standardised, enhanced Cryptosporidium exposure questionnaire was administered to all cases of cryptosporidiosis after the incident. Stool samples, water testing, microscopy slides and rabbit gut contents positive for Cryptosporidium were typed at the Cryptosporidium Reference Unit, Singleton Hospital, Swansea. Twenty-three people were microbiologically linked to the incident although other evidence suggests an excess of 422 cases of cryptosporidiosis above baseline. Most were adult females; unusually for cryptosporidiosis there were no affected children identified under the age of 5 years. Water consumption was possibly higher than in national drinking water consumption patterns. Diarrhoea duration was negatively correlated to distance from the water treatment works where the contamination occurred. Oocyst counts were highest in water storage facilities. This outbreak is the first caused by C. cuniculus infection to have been noted and it has conclusively demonstrated that this species can be a human pathogen. Although symptomatically similar to cryptosporidiosis from C. parvum or C. hominis, this outbreak has revealed some differences, in particular no children under 5 were identified and females were over-represented. These dissimilarities are unexplained although we postulate possible explanations. PMID:24642431

  7. Hyperbaric oxygen therapy as additional treatment in deep sternal wound infections – a single center's experience

    PubMed Central

    Bryndza, Magdalena; Chrapusta, Anna; Kobielska, Ewa; Kapelak, Bogusław; Grudzień, Grzegorz

    2016-01-01

    Introduction Deep sternal wound infection (DSWI) is one of the most serious complications after cardiac surgery procedures, observed in 5% of patients. Current standard medical therapy for DSWI includes antibiotics, surgical debridement, resuturing or negative pressure wound therapy (NPWT). Unfortunately, in some cases these methods are insufficient, and additional therapeutic options are needed. Aim To assess the effects and usefulness of additional hyperbaric oxygen therapy (HBO2) in patients with DSWI after cardiac surgery procedures. Material and methods A retrospective analysis of 10 patients after cardiac surgery who developed DSWI in the period 2010–2012 was performed. After 3 months of ineffective conventional therapy including targeted antibiotic, surgical sternal debridement and NPWT, patients were qualified for additional HBO2 therapy. A total of 20 sessions of HBO2 therapy were performed, each 92 minutes long. Results After 4 weeks of HBO2 treatment, 7 patients presented complete wound healing with fibrous scar formation. One patient was qualified for the another cycle of HBO2 therapy with 20 additional sessions, and complete wound healing was observed. In 2 cases, after 5 and 19 sessions, HBO2 was interrupted because of improper qualifications. Conclusions The HBO2 as an additional therapy in DSWI was successful in 80% of cases, and no complications were observed. However, due to the small number of published studies with a small number of patients, randomized, clinical trials are needed to assess the clinical results of HBO2 in DSWI after cardiac surgery procedures. PMID:27785131

  8. 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

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

    PubMed

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

    2009-12-01

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

  10. 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

  11. 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.

  12. 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

  13. Identification of novel Cryptosporidium species in aquarium fish.

    PubMed

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

    2010-11-24

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

  14. 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

  15. 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.

  16. 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

  17. Additive effect of ketoconazole and octreotide in the treatment of severe adrenocorticotropin-dependent hypercortisolism.

    PubMed

    Vignati, F; Loli, P

    1996-08-01

    Over the last few years ketoconazole and octreotide have been employed in the treatment of pituitary-dependent or ectopic Cushing's syndrome. In four patients (two men and two women, aged 25-64 yr) with severe ACTH-dependent hypercortisolism in whom medical treatment with ketoconazole showed limited effectiveness and/or tolerability, we tried the association with octreotide. In all patients ketoconazole (200-1000 mg) induced a marked decrease in urinary free cortisol (UFC) excretion, but normalization could not be achieved. After ketoconazole discontinuation, three patients received octreotide alone (300-1500 micrograms/day, sc). This drug caused a dramatic decrease in UFC excretion, although not normalization; in all patients, escape from treatment occurred. Combined treatment was carried out for 10-180 days. Urinary cortisol excretion normalized and remained steadily within normal limits in three of four patients in whom normal UFC excretion had never been attained with both single drug regimens; in the fourth patient, UFC excretion decreased to levels lower than those achieved with ketoconazole or octreotide alone. The association with octreotide allowed a reduction in the daily dose of ketoconazole in three patients. Consistent with the steady reduction of cortisol production, a striking clinical improvement occurred in all patients after starting combined treatment. The normalization of UFC in three of four patients treated with both agents suggests that this approach may be useful in the long term treatment of severe forms of hypercortisolism of both pituitary and ectopic origin. In contrast to the limited effectiveness of each drug taken singularly at the same or higher doses, the association of the two drugs had an additive effect in the attainment of normal urinary cortisol excretion.

  18. Additive Effect of Neutralizing Antibody and Antiviral Drug Treatment in Preventing Virus Escape and Persistence

    PubMed Central

    Seiler, Peter; Senn, Beatrice M.; Klenerman, Paul; Kalinke, Ulrich; Hengartner, Hans; Zinkernagel, Rolf M.

    2000-01-01

    Poorly cytopathic or noncytopathic viruses can escape immune surveillance and establish a chronic infection. Here we exploited the strategy of combining antiviral drug treatment with the induction of a neutralizing antibody response to avoid the appearance of neutralization-resistant virus variants. Despite the fact that H25 immunoglobulin transgenic mice infected with lymphocytic choriomeningitis virus mounted an early neutralizing antibody response, the virus escaped from neutralization and persisted. After ribavirin treatment of H25 transgenic mice, the appearance of neutralization-resistant virus was prevented and virus was cleared. Thus, the combination of virus-neutralizing antibodies and chemotherapy efficiently controlled the infection, whereas each defense line alone did not. Similar additive effects may be unexpectedly efficient and beneficial in humans after infections with persistent viruses such as hepatitis C virus and hepatitis B virus and possibly human immunodeficiency virus. PMID:10846070

  19. Treatment of a dilute waste oil emulsion by chemical addition (CA)-dissolved air flotation (DAF)

    SciTech Connect

    Reed, B.E.; Carriere, P.; Zhu, X.; Lorkowski, T.

    1995-12-31

    Treatment of wastewater from aluminum rolling mill operations is an inherent problem in the aluminum fabrication industry. In this study, wastewater from the manufacturer`s processes was transferred to two holding ponds having a total capacity of about 5 million gallons and a detention time of about 10 days. In the holding ponds, free oil was allowed to rise to the surface where it was periodically removed. Wastewater from the holding ponds was withdrawn from about a depth of 9 ft. and used as influent to a variety of technologies. In this paper, results from the chemical addition (CA)-dissolved air flotation (DAF) portion of the treatability study are presented.

  20. 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...

  1. Performance of basaltic dust issued from an asphaltic plant as a flocculant additive for wastewater treatment.

    PubMed

    Ramírez Zamora, R M; Chávez Mejia, A; Domínguez Mora, R; Durán Moreno, A

    2004-01-01

    The feasibility of using basaltic dust as a flocculant additive or coagulant aid for wastewater treatment was assessed in this research. The experimental study was divided into two stages: 1) physicochemical characterisation of the basaltic dust by applying standardised techniques, and 2) evaluation of this material as flocculant additive for the coagulation-flocculation of wastewater treated for reuse. Coagulation-flocculation experiments were carried out in the laboratory with a mixture of industrial and municipal wastewater samples collected from two points of the final discharge of the Mexico City sewerage system. Aluminium sulphate and lime were used as coagulants and the basaltic dust as flocculant additive, by applying the jar-test technique. The results of the corrosivity, reactivity, explosiveness, toxicity, inflammability and biological risk tests indicated that this material is classified as a non-hazardous waste (according to the Mexican legislation, NOM-052-ECOL-1993). The density, oxide content and particle size values of basaltic dust were similar to those reported for the flocculant additive denominated activated silica. The jar test results showed a positive effect of basaltic dust over the effluent and sludge qualities, to the extent that coagulant doses can be reduced 30% (from 150 mg/L to 110 mg/L of Al2(SO4)3).

  2. 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

  3. 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

  4. 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... reasonably be prevented through optimizing treatment plant design, operation, and maintenance. (ii) The... performance. The State may approve Cryptosporidium treatment credit for drinking water treatment...

  5. 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... reasonably be prevented through optimizing treatment plant design, operation, and maintenance. (ii) The... performance. The State may approve Cryptosporidium treatment credit for drinking water treatment...

  6. 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... reasonably be prevented through optimizing treatment plant design, operation, and maintenance. (ii) The... performance. The State may approve Cryptosporidium treatment credit for drinking water treatment...

  7. 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...

  8. 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...

  9. Raman spectroscopic evaluation of meat batter structural changes induced by thermal treatment and salt addition.

    PubMed

    Herrero, A M; Carmona, P; López-López, I; Jiménez-Colmenero, F

    2008-08-27

    Raman spectroscopy, texture, proximate composition, and water binding analysis were carried out to evaluate the effect of thermal treatment and/or salt addition to meat batter. For this purpose, different meat batters were elaborated: control meat batter (no salt) and meat batters with low (1.0%) and high (2.5%) NaCl content with and without thermal treatment (70 degrees C/30 min). Increase (P < 0.05) in penetration force and hardness upon heating was observed. Results also showed hardness increasing (P < 0.05) as a function of salt addition in heated meat batter. Raman spectroscopy analysis revealed a significant (P < 0.05) decrease in alpha-helix content accompanied by an increase (P < 0.05) in beta-sheets resulting from heating. Significant (P < 0.05) correlations were found between these secondary structural changes in meat proteins and water binding and textural properties of meat batter. In this way, a significant correlation was found between beta-sheets, salt content, hardness, and chewiness in heated samples.

  10. 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.

  11. 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...

  12. 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 ...

  13. ANIMAL SOURCE IDENTIFICATION USING A CRYPTOSPORIDIUM DNA CHARACTERIZATION TECHNIQUE

    EPA Science Inventory

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

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

    PubMed

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

    2014-10-01

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

  15. 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.

  16. 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.

  17. 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.

  18. 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

  19. 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

  20. 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

  1. 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

  2. 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

  3. 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).

  4. 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

  5. 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

  6. 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

  7. 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

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

    PubMed

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

    2016-10-01

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

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

    PubMed

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

    2016-10-01

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

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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

  15. Effect of the addition of calcium hydroxide on the hydrothermal-mechanochemical treatment of Eucalyptus.

    PubMed

    Ishiguro, Maki; Endo, Takashi

    2015-02-01

    The effect of Ca(OH)2 addition on optimization of hydrothermal-mechanochemical pretreatment, which combines hydrothermal and milling treatments, was examined. The highest glucose yield of 90% was achieved in the ball-milled specimen previously treated at 170°C in the presence of 20% Ca(OH)2 per substrate weight. The specific surface area of the substrate was closely correlated with glucose yield, and a larger specific surface area was obtained when treating the specimen at 170°C in the presence of Ca(OH)2 compared to treatment at 170°C without Ca(OH)2. Although the Ca(OH)2-treated specimen was relatively unaffected by delignification, the cleavage of the ester bonds between lignin and hemicellulose was confirmed by FT-IR. This suggests that Ca(OH)2 weakens the substrate structure by loosening the bonds between lignin and hemicellulose as the mechanism to increase the specific surface area regardless of the high lignin content, facilitating the fibrillation of fibers with mechanical milling.

  16. 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

  17. 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

  18. 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

  19. Upper respiratory tract infection caused by Cryptosporidium baileyi in three mixed-bred falcons (Falco rusticolus x Falco cherrug).

    PubMed

    van Zeeland, Y R A; Schoemaker, N J; Kik, M J L; van der Giessend, J W B

    2008-06-01

    Three mixed-bred raptors (Falco rusticolus x Falco cherrug) from a German falcon breeder were presented with a history of respiratory distress. In one bird a laryngeal stridor was noted, and oral examination revealed an epiglottal swelling. In the other two birds, nasal discharge and sneezing were the main clinical symptoms. Nasal flushing samples and biopsies were collected for pathologic, bacteriologic, and parasitologic examination. Results confirmed a cryptosporidial infection. Polymerase chain reaction (PCR) and DNA analysis identified the causative agent to be Cryptosporidium baileyi. No cryptosporidia were detected in fecal samples, indicating the infection was confined to the respiratory system. Analysis of prey animals (pigeons, quail) failed to identify the source of infection. Treatment was initiated with paromomycin in all three birds, whereas in two birds an additional therapy with azithromycin was given. However, no clinical improvement was seen after several weeks of treatment, and the birds either died or were euthanatized. To the authors' knowledge, these are the first confirmed cases of disease caused by cryptosporidia in the order of Falconiformes.

  20. 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...

  1. 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.

  2. 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

  3. 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...

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

    PubMed

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

    2011-11-01

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

  5. Continuous culture of Cryptosporidium parvum using hollow fiber technology.

    PubMed

    Morada, Mary; Lee, Sangun; Gunther-Cummins, Leslie; Weiss, Louis M; Widmer, Giovanni; Tzipori, Saul; Yarlett, Nigel

    2016-01-01

    Diarrheal disease is a leading cause of pediatric death in economically low resource countries. Cryptosporidium spp. are the second largest member of this group and the only member for which no treatment exists. One of the handicaps to developing chemotherapy is the lack of a reproducible long-term culture method permitting in vitro drug screening beyond 48 h. We have adapted the well-established hollow fiber technology to provide an environment that mimics the gut by delivering nutrients and oxygen from the basal layer upwards while allowing separate redox and nutrient control of the lumen for parasite development. Using this technique, oocyst production was maintained for >6 months, producing approximately 1×10(8)oocysts ml(-1)day(-1), compared with 48 h with a yield of 1×10(6)oocysts ml(-1) in two-dimensional cultures. Oocysts, after 4 and 20 weeks in culture, produced a chronic infection in a TCR-α-deficient mouse model. In vivo infectivity of oocysts was confirmed using oocysts from a 6 week culture in a dexamethasone immunosuppressed mouse model. PMID:26341006

  6. 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

  7. 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

  8. 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

  9. 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

  10. 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

  11. 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.

  12. 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.

  13. High-dose nedocromil sodium as an addition to inhaled corticosteroids in the treatment of asthma.

    PubMed

    O'Hickey, S P; Rees, P J

    1994-08-01

    In a double-blind placebo-controlled trial nedocromil sodium in a dose of 8 mg four times daily or matching placebo was added to the treatment of 29 asthmatic patients. All patients were taking inhaled corticosteroids in a dose of up to 1000 micrograms daily. The trial agents were given for 6 weeks after a 2-week run-in period. Twenty-four patients completed the study, three withdrew because of adverse effects, two on placebo. Daytime asthma symptoms were significantly reduced on nedocromil compared to placebo (-0.46 vs. +0.09, P = 0.03). Night-time asthma and morning tightness were not changed significantly. Bronchodilator use in the night and day were lower on nedocromil but the differences were not significant. Morning peak flow rates were higher on nedocromil (+22.2 vs. +0.08, P = 0.06) and physicians opinions of overall effectiveness favoured nedocromil (U = 35.0, P = 0.04). These results confirm that nedocromil sodium may be a useful addition in asthma to low to medium doses of inhaled corticosteroids. The effects of 32 mg nedocromil daily were comparable to previous reports with lower doses.

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

    PubMed

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

    2011-03-01

    Cryptosporidium is a coccidian parasite that is prevalent worldwide, some species of which cause morbidity in both immunocompromised and immunocompetent individuals. The prevalence and predictors of Cryptosporidium infection, and its effect on nutritional status, have recently been explored among 276 children (141 boys and 135 girls, aged 2-15 years) in aboriginal (Orang Asli) villages in the Malaysian state of Selangor. Faecal smears were examined by the modified Ziehl-Neelsen staining technique while socio-economic data were collected using a standardized questionnaire. Nutritional status was assessed by anthropometric measurements. Cryptosporidium infection, which was detected in 7.2% of the aboriginal children, was found to be significantly associated with low birthweight (≤2.5 kg), being part of a large household (with more than seven members) and prolonged breast feeding (>2 years). The output of a binary logistic regression confirmed that large household size was a significant predictor of Cryptosporidium infection (giving an odds ratio of 2.15, with a 95% confidence interval of 1.25-5.02). Cryptosporidium infection is clearly a public-health problem among the aboriginal children of Selangor, with person-to-person the most likely mode of transmission. PMID:21396250

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

    PubMed

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

    2011-03-01

    Cryptosporidium is a coccidian parasite that is prevalent worldwide, some species of which cause morbidity in both immunocompromised and immunocompetent individuals. The prevalence and predictors of Cryptosporidium infection, and its effect on nutritional status, have recently been explored among 276 children (141 boys and 135 girls, aged 2-15 years) in aboriginal (Orang Asli) villages in the Malaysian state of Selangor. Faecal smears were examined by the modified Ziehl-Neelsen staining technique while socio-economic data were collected using a standardized questionnaire. Nutritional status was assessed by anthropometric measurements. Cryptosporidium infection, which was detected in 7.2% of the aboriginal children, was found to be significantly associated with low birthweight (≤2.5 kg), being part of a large household (with more than seven members) and prolonged breast feeding (>2 years). The output of a binary logistic regression confirmed that large household size was a significant predictor of Cryptosporidium infection (giving an odds ratio of 2.15, with a 95% confidence interval of 1.25-5.02). Cryptosporidium infection is clearly a public-health problem among the aboriginal children of Selangor, with person-to-person the most likely mode of transmission.

  16. 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

  17. 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

  18. 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.

  19. 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

  20. [Spontaneous Cryptosporidium infection in weaned rabbits].

    PubMed

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

    1996-12-01

    The first occurrence of Cryptosporidium parvum Tyzzer, 1912 in broiler rabbits in the Czech Republic is reported. The protozoon was determined on the basis of morphometrical parameters of oocysts and of localization of endogenous developmental stages. The dynamics of natural Cryptosporidium infection was studied in a group of 72 young rabbits after weaning (their age ranging from 23-33 to 82-92 days) obtained from six large flocks and used in a feeding experiment. C. parvum was found in rabbits from four farms (Tab. I). Animals under observation were divided into 9 subgroups according to the genotype (Hyla 2000, California White, crosses of New Zealand x California, New Zealand White, Cunistar and Zika) as well as according to the farm of origin. The animals were housed in 28 cages under the conditions of two-floor cage technology. The upper floor consisted of cages housing three head, the lower floor two head each. The animals were fed ad libitum with commercial feed mixture (till the average age of 64.days supplemented with Robenidin as coccidiostat). During the first 10 days of observation pooled samples of droppings from each cage were examinated by flotation-centrifugation method according to Breza (1957) and Pavlásek (1991) in the intervals of three to four days, later one-week intervals. Post mortem scrapings from mucous epithelium taken from young rabbits were examinated (to reveal endogenous developmental stages of C. parvum) together with digesta (to detect oocysts of the protozoon) taken from the full length of the small intestine using method of native preparations and Giemsa stain. In one 37-day dead animal the small and large intestines were examined histologically. The maximum number of young rabbits infected with C. parvum were 30-40 and 33-43 days old (Fig. 1). In animals of this age category the oocysts of the protozoon were found in pooled samples in 11 and 12 cages (39.3 and 42.9%) from totally 28 cages under study. In rabbits of more than 50

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

    PubMed

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

    2016-01-15

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

  2. 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.

  3. Dissection of the hierarchy and synergism of the bile derived signal on Cryptosporidium parvum excystation and infectivity.

    PubMed

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

    2012-10-01

    Bile salts have been identified as an important trigger for excystation of Cryptosporidium oocysts but the hierarchy or synergism of this signal in relation to other triggers involved in excystation is poorly understood. In addition to excystation, bile salts have also been reported to increase the invasiveness of sporozoites within in vitro culture, possibly by affecting the secretory pathway via modification of intracellular calcium signalling. Nevertheless, incorporation of bile or bile salts into in vitro assays is not universal, with recent reports of negative effects on parasite growth. Here we report that bile and sodium taurocholate significantly affect both excystation rate and parasite in vitro growth. We demonstrate that their effect on excystation is dose, time and pre-treatment temperature dependent, while increases in parasite replication appear to be associated with modulation of parasite intracellular calcium and increased host cell susceptibility to infection. Notably, we illustrate that bile has a significant effect on host cells and can be cytotoxic at concentrations not much higher than those currently used for in vitro assays. This work should assist with more rational design of in vitro culture systems, with significant considerations for assay format when incorporating bile or bile salts as an excystation trigger. PMID:22894830

  4. Occurrence and molecular characterization of Cryptosporidium spp. in mammals and reptiles at the Lisbon Zoo.

    PubMed

    Alves, Margarida; Xiao, Lihua; Lemos, Vanessa; Zhou, Ling; Cama, Vitaliano; da Cunha, Margarida Barão; Matos, Olga; Antunes, Francisco

    2005-09-01

    The presence of Cryptosporidium parasites in mammals and reptiles kept at the Lisbon Zoo was investigated. A total of 274 stool samples were collected from 100 mammals and 29 reptiles. The species and genotype of the isolates identified by light microscopy were determined by nested PCR and sequence analysis of a fragment of the small subunit rRNA gene. Cryptosporidium oocysts were found in one black wildebeest (Connochaetes gnou), one Prairie bison (Bison bison bison) and in one Indian star tortoise (Geochelone elegans). The PCR and sequence analysis of these three isolates showed that those excreted by the Prairie bison were Cryptosporidium mouse genotype, those from the black wildebeest were from a new Cryptosporidium genotype and those infecting the Indian star tortoise were Cryptosporidium tortoise genotype. The present work reports a new Cryptosporidium genotype in a black wildebeest and the first finding of the Cryptosporidium mouse genotype in a ruminant.

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

    PubMed Central

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

    2007-01-01

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

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

    PubMed

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

    2007-10-01

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

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

    PubMed

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

    2010-02-01

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

  8. 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

  9. 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.

  10. A foodborne outbreak of Cryptosporidium hominis infection.

    PubMed

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

    2009-03-01

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

  11. Viability of preserved Cryptosporidium baileyi oocysts

    PubMed Central

    Surl, Chan-Gu; Kim, Se-Min

    2003-01-01

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

  12. 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.

  13. 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.

  14. 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...

  15. 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

  16. 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.

  17. 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

  18. 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

  19. 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

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

    PubMed

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

    2015-02-01

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

  1. 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.

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

    PubMed

    Robinson, Guy; Chalmers, Rachel M

    2012-10-01

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

  3. Adaptation and immunogenicity of Cryptosporidium parvum to immunocompetent mice.

    PubMed

    Matsuo, Tomohide; Tsuge, Yasuko; Umemiya-Shirafuji, Rika; Fujino, Takashi; Matsui, Toshihiro

    2014-03-01

    The adaptation and immunogenisity of Cryptosporidium parvum isolated from Siberian chipmunks (SC1 strain) in immunocompetent (ICR) mice were examined. The oocysts were received to the severe combined immunodeficiency (SCID) mice by repeated passage. The oocysts collected from the 18th SCID mice were inoculated to 5 ICR mice. The mice began to shed oocysts from 6 days after inoculation, the patency was 5 days, and the maximum oocysts per gram of feces (OPG) value was 10(4). The maximum of OPG value was gradually increased by successive passage, and finally that in the 22nd mice reached 10(6) (patency: 11 days). It is considered that these results indicate completion of their adaptation to ICR mice. To examine the immunogenicity of C. parvum to ICR mice, 8 groups of 5 mice each were inoculated with 1.3 × 10(6) oocysts of SC1 strain, which were collected after adaptation to SCID mice. All groups shed oocysts from 6th day, and their patency was from 8 to 12 days. On the 21st day after the primary infection, these mice were challenged with 1.3 × 10(6) oocysts. No oocysts shed from any groups, although 2 control groups shed oocysts from the 6th day, and their OPG values were more than 10(6). These results suggest that this strain has strong immunogenicity against ICR mice. Therefore, the immunological healthy mice were considered a useful experimental model to investigate immunological and drug treatments in the strain of C. parvum.

  4. Quantitation of Giardia cysts and Cryptosporidium oocysts in fecal samples by direct immunofluorescence assay.

    PubMed Central

    Xiao, L; Herd, R P

    1993-01-01

    The lack of quick, simple, and sensitive quantitative tests has impeded studies on infection patterns and treatment of Giardia spp. and Cryptosporidium spp. A quantitative direct immunofluorescence assay (FA) using a commercial FA kit was developed and evaluated. Recovery rates of the FA for Cryptosporidium oocysts in calf feces seeded with 1,000, 10,000, 100,000, and 1,000,000 oocysts per g were 14.8, 40.8, 84.2, and 78.2%, respectively. Interassay coefficients of variation were 10.6 to 47.1%. Recovery rates of the FA for Giardia cysts in feces seeded with 1,000, 10,000, and 100,000 cysts per g were 76.4, 96.9, and 89.6%, respectively. Interassay coefficients of variation were 7.4 to 22.1%. By comparison, recovery rates of Giardia cyst by sucrose gradient flotation were only 20.5, 51.2, and 42.9%, respectively. Counts of cysts-per-gram obtained by sucrose gradient flotation with samples from calves, lambs, and ewes were only 49.1 to 54.8% of those obtained by the FA. Zinc sulfate flotation detected only 36.4% of infections when there were < or = 1,000 cysts per g. The quantitative FA offers a useful technique for epidemiological and control studies of these two parasites. PMID:8263179

  5. 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

  6. 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

  7. 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

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

    PubMed

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

    2003-05-01

    Cryptosporidium parvum represents a challenge to the water industry and a threat to public health. In this study, we developed a cell culture-quantitative PCR assay to evaluate the inactivation of C. parvum with disinfectants. The assay was validated by using a range of disinfectants in common use in the water industry, including low-pressure UV light (LP-UV), ozone, mixed oxidants (MIOX), and chlorine. The assay was demonstrated to be reliable and sensitive, with a lower detection limit of a single infectious oocyst. Effective oocyst inactivation was achieved (>2 log(10) units) with LP-UV (20 mJ/cm(2)) or 2 mg of ozone/liter (for 10 min). MIOX and chlorine treatments of oocysts resulted in minimal effective disinfection, with <0.1 log(10) unit being inactivated. These results demonstrate the inability of MIOX to inactivate Cryptosporidium. The assay is a valuable tool for the evaluation of disinfection systems for drinking water and recycled water.

  9. Antibodies Fused to Innate Immune Molecules Reduce Initiation of Cryptosporidium parvum Infection in Mice▿

    PubMed Central

    Imboden, Michael; Riggs, Michael W.; Schaefer, Deborah A.; Homan, E. Jane; Bremel, Robert D.

    2010-01-01

    At present no completely effective treatments are available for Cryptosporidium parvum infections in humans and livestock. Based on previous data showing the neutralizing potential of a panel of monoclonal antibodies developed against C. parvum, and based on the fact that innate immune peptides and enzymes have anticryptosporidial activity, we engineered several of these antibodies into antibody-biocide fusion proteins. We hypothesized that the combination of high-affinity antibody targeting with innate immune molecule-mediated killing would result in a highly effective new antiprotozoal agent. To test this hypothesis, we expressed antibody-biocide fusion proteins in a mammalian cell culture system and used the resulting products for in vitro and in vivo efficacy experiments. Antibody-biocide fusion proteins efficiently bound to, and destroyed, C. parvum sporozoites in vitro through a membrane-disruptive mechanism. When antibody-biocide fusion proteins were administered orally to neonatal mice in a prophylactic model of cryptosporidiosis, the induction of infection was reduced by as much as 81% in the mucosal epithelium of the gut, as determined on the basis of histopathological scoring of infectious stages. Several versions of antibody fusion proteins that differed in antigen specificity and in the biocide used had strong inhibitory effects on the initiation of infection. The results lay the groundwork for the development of a new class of antimicrobials effective against Cryptosporidium. PMID:20086143

  10. 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

  11. 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

  12. 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...

  13. 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...

  14. 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 ...

  15. 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...

  16. 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...

  17. 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...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  19. Biofilms Reduce Solar Disinfection of Cryptosporidium parvum Oocysts

    PubMed Central

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

    2012-01-01

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

  20. 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...

  1. 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...

  2. DIFFERENTIATING HUMAN FROM ANIMAL ISOLATES OF CRYPTOSPORIDIUM PARVUM

    EPA Science Inventory

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

  3. 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...

  4. 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

  5. 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...

  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. Quantitative risk assessment of Cryptosporidium species infection in dairy calves.

    PubMed

    Nydam, D V; Mohammed, H O

    2005-11-01

    Cryptosporidium parvum is a zoonotic protozoan that infects many different mammals including cattle and humans. Cryptosporidiosis has become a concern for dairy producers because of the direct losses due to calves not performing well and the potential for environmental contamination with C. parvum. Identifying modifiable control points in the dynamics of infection in dairy herds will help identify management strategies that mitigate its risk. The quantitative risk assessment approach provides estimates of the risk associated with these factors so that cost-effective strategies can be implemented. Using published data from epidemiologic studies and a stochastic approach, we modeled the risk that C. parvum presents to dairy calves in 2 geographic areas: 1) the New York City Watershed (NYCW) in southeastern New York, and 2) the entire United States. The approach focused on 2 possible areas of exposure--the rearing environment and the maternity environment. In addition, we evaluated the contribution of many risk factors (e.g., age, housing, flies) to the end-state (i.e., total) risk to identify areas of intervention to decrease the risk to dairy calves. Expected risks from C. parvum in US dairy herds in rearing and maternity environments were 41.7 and 33.9%, respectively. In the NYCW, the expected risks from C. parvum in the rearing and maternity environments were 0.36 and 0.33%, respectively. In the US scenarios, the immediate environment contributed most of the risk to calves, whereas in the NYCW scenario, it was new calf infection. Therefore, within the NYCW, risk management activities may be focused on preventing new calf infections, whereas in the general US population, cleaning of calf housing would be a good choice for resource allocation. Despite the many assumptions inherent with modeling techniques, its usefulness to quantify the likelihood of risk and identify risk management areas is illustrated.

  8. 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…

  9. Clinical Psychopharmacology Update: Additional Safety Concerns for Using Varenicline (Chantix) for Smoking Cessation Treatment.

    PubMed

    Tobin, Thomas J; Tobin, Mary L

    2015-01-01

    Smoking cessation remains a positive therapeutic goal and should be encouraged for the millions of individuals who continue to smoke and struggle to quit. While psychiatric nurses should encourage patients to start or continue smoking cessation therapies, they must be aware of the additional safety concerns relating to the use of varenicline (Chantix). Research published subsequent to the last clinical update in this journal (Tobin, 2007 ) has prompted additional warnings from the Food and Drug Administration concerning varenicline for smoking cessation therapy. In particular, clinicians need to be aware of increased concerns about varenicline's association with neuropsychiatric side effects, seizures, and alcohol interactions. PMID:26514264

  10. 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.

  11. Laser heat treatment of aerosol-jet additive manufactured graphene patterns

    NASA Astrophysics Data System (ADS)

    Jabari, Elahe; Toyserkani, Ehsan

    2015-09-01

    In this article, a laser processing protocol for heat treatment of micro-scale printed graphene patterns is developed, and the results are compared with the counterpart results obtained by the conventional heat treatment process carried out in a furnace. A continuous-wave Erbium fiber laser is used to enhance electrical properties of the aerosol-jet printed graphene patterns through removing solvents and a stabilizer polymer. The laser power and the process speed are optimized to effectively treat the printed patterns without compromising the quality of the graphene flakes. Furthermore, a heat transfer model is developed and its results are utilized to optimize the laser treatment process. It is found that the laser heat treatment process with a laser speed of 0.03 mm s-1, a laser beam diameter ~50 μm, and a laser power of 10 W results in pure graphene patterns with no excessive components. The ratio of D to G bands ({{I}\\text{D}}/{{I}\\text{G}}) in Raman graph of the laser treated pure graphene, which is an indicator of the level of the active defects in graphene structures, is 0.52. The laser treated pure graphene structures also have a C/O ratio and an electrical resistivity of ~4.5 and 0.022 Ω cm, respectively. These values are fairly comparable with the results of samples treated in a furnace. The results suggest that the laser processing has the capability of removing stabilizer polymers and solvents through a localized moving heat source, which is preferable for flexible electronics with low working temperature substrates.

  12. 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

  13. 42 CFR 412.107 - Special treatment: Hospitals that receive an additional update for FYs 1998 and 1999.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... Inpatient Operating Costs § 412.107 Special treatment: Hospitals that receive an additional update for FYs... section if it meets all of the following criteria: (1) Definition. The hospital is not a Medicare...) of this section for their cost reporting periods beginning in FY 1995 is less than the...

  14. 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

  15. 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

  16. Treatment of Scumming Effects of Pottery Clay by Sodium Carbonate Addition

    NASA Astrophysics Data System (ADS)

    Wasanapiarnpong, T.; Thueploy, A.; Nilpairach, S.; Arayaphong, D.

    2011-10-01

    Earthenware pottery products made by using red plastic clay in Ratchaburi province of Thailand and fired at 850-1000 °C, always shows some blemishes, caused by scumming on the surface. This scumming contains calcium sulfate, contaminated in the raw clay as gypsum form. The addition of barium carbonate is a suggested solution to prevent this white stain. However, it is difficult for barium carbonate to spread throughout the clay so that it takes a long time to complete the reaction. This research aims to find the solution by using sodium carbonate as an alternative chemical. Sodium carbonate was mixed in the clay at 1wt% dissolved in distilled water controlled the moisture at 22 % by wet weight. The mixture was kneaded and aged for 24 h, then formed, dried and fired at 850-950 °C. The types and quantities of ion in mixed clay and deposited on the surface product were determined after drying. It was found that the white stain areas were diminished, as same as the result from the addition of barium carbonate. Moreover, the sample after firing at 950 °C had lower water absorption as 12.22 %, higher three point bending strength as 32.53 MPa when compared to the addition of barium carbonate, which had higher water absorption as 15.58 % and lower three point bending strength as 25.25 MPa.

  17. Nutrient removal in wastewater treatment high rate algal ponds with carbon dioxide addition.

    PubMed

    Park, J B K; Craggs, R J

    2011-01-01

    The influence of CO2 addition to high rate algal ponds (HRAPS) on nitrogen removal was investigated using two pilot-scale HRAPs operated with different hydraulic retention times (HRT: 4 and 8 days), and was compared to the nitrogen removal by the 8-day HRT pond before CO2 addition was installed. Nitrogen balances were calculated by partitioning total nitrogen into organic and inorganic nitrogen (NH4+-N and NO3--N), and by separation of the organic nitrogen into particulate (PON) and dissolved organic nitrogen (DON). PON was further divided into algal organic nitrogen (AON) and bacteria organic nitrogen (BON) to investigate nitrogen mass flow in the HRAPS. This research shows that the proportion of algae in the algal/bacterial biomass in the longer 8-day HRT HRAP8d (55.6%) was appreciably lower than that in the shorter 4-day HRT HRAP4d (80.5%) when CO2 was added to control the maximum pH to <8.0 during the summer. Higher bacterial biomass in the longer 8-day HRT HRAP corresponded with higher nitrification rates, indicating that the longer 8-day HRT in the summer was detrimental for two reasons: lower algal productivity and increased nitrogen loss through nitrification/denitrification. Overall nitrogen removal of approximately 60% in the HRAPS with CO2 addition was mainly achieved by algal assimilation followed by sedimentation in the settling unit.

  18. 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

  19. 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

  20. Addition of a magnetite layer onto a polysulfone water treatment membrane to enhance virus removal.

    PubMed

    Raciny, I; Zodrow, K R; Li, D; Li, Q; Alvarez, P J J

    2011-01-01

    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 membranes coated with nano-Fe3O4 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 coloured water. Further research is needed to reduce the loss of water flux caused by coating.

  1. 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.

  2. Addition of a magnetite layer onto a polysulfone water treatment membrane to enhance virus removal.

    PubMed

    Raciny, I; Zodrow, K R; Li, D; Li, Q; Alvarez, P J J

    2011-01-01

    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 membranes coated with nano-Fe3O4 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 coloured water. Further research is needed to reduce the loss of water flux caused by coating. PMID:21977659

  3. 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.

  4. 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

  5. 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).

  6. 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

  7. 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

  8. 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

  9. 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...

  10. Synthesis, in Vitro Evaluation and Cocrystal Structure of 4-Oxo-[1]benzopyrano[4,3-c]pyrazole Cryptosporidium parvum Inosine 5′-Monophosphate Dehydrogenase (CpIMPDH) Inhibitors

    PubMed Central

    2015-01-01

    Cryptosporidium inosine 5′-monophosphate dehydrogenase (CpIMPDH) has emerged as a therapeutic target for treating Cryptosporidium parasites because it catalyzes a critical step in guanine nucleotide biosynthesis. A 4-oxo-[1]benzopyrano[4,3-c]pyrazole derivative was identified as a moderately potent (IC50 = 1.5 μM) inhibitor of CpIMPDH. We report a SAR study for this compound series resulting in 8k (IC50 = 20 ± 4 nM). In addition, an X-ray crystal structure of CpIMPDH·IMP·8k is also presented. PMID:25474504

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

    PubMed

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

    2012-07-01

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

  12. 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.

  13. 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

  14. 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.

  15. [Up-to-date drug treatment of disseminated lung cancer--which other drugs are available in addition to conventional cytotoxic agents?].

    PubMed

    Koivunen, Jussi; Knuuttila, Aija; Mali, Pekka

    2016-01-01

    In addition to conventional cytotoxic agents, novel drug treatments have in the last few years been introduced for the treatment of non-small cell lung cancer. Whereas some of the novel treatments have brought significant improvement in treatment outcome, the benefit brought about by the treatment has in some cases been quite small in comparison with the costs and adverse effects. In the present review we explore the goals of drug treatments of disseminated lung cancer, assessment of therapeutic benefits as well as most significant research results of novel drug treatments of the lastfew years In addition, we evaluate the effect of the novel drug treatments on Finnish treatment practices.

  16. 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.

  17. Comparison of assays for sensitive and reproducible detection of cell culture-infectious Cryptosporidium parvum and Cryptosporidium hominis in drinking water.

    PubMed

    Johnson, Anne M; Giovanni, George D Di; Rochelle, Paul A

    2012-01-01

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

  18. 77 FR 57545 - Long Term 2 Enhanced Surface Water Treatment Rule: Public Meeting on Monitoring Data Analysis...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-18

    ... enhanced analytical methods. The third topic is the effectiveness of Escherichia coli as a screen to... 2006 (71 FR 654; January 5, 2006), is to reduce disease incidence associated with Cryptosporidium and... treatment that may be needed based on the occurrence of Cryptosporidium or E. coli in their source...

  19. Preparation and characterization of ceramic products by thermal treatment of sewage sludge ashes mixed with different additives.

    PubMed

    Merino, Ignacio; Arévalo, Luis F; Romero, Fernando

    2007-01-01

    The study of the ceramic characteristics of sludge ashes, alone or mixed with additives (kaolin, montmorillonite, illitic clay, powdered flat glass) includes characterization of additives, preparation of probes (dry or wet mixed), thermal treatment (up to 1200 degrees C, except melting or deformation) and control (densities, compressive strengths and water absorption). Thermal treatment increases the density and compressive strength of probes (both parameters go through maxima, with later decreases) and decreases the absorption of water. The densification is also revealed by the evolution of the ratio of decrease of volume/loss of mass. The maximum values of compressive strengths were obtained for 25% of illitic clay, montmorillonite and glass powder. Densification concerning probes with sludge ashes alone does not occur with kaolin. Experimental data were adjusted to exponential relationships between compressive strengths and densities for every composition, and also to a general equation for all probes. The apparent density obtained was adjusted to a non-linear dependence with temperature, leading to a maximum in density and permitting calculating the temperature of occurrence of this maximum. The adjustment was not possible for probes containing kaolin, requiring presumably higher temperatures to densify. Water absorption has low values for ashes or kaolin probes, intermediate values for illite and powdered flat glass probes and high values for montmorillonite probes. Excepting with kaolin, ceramic materials with better characteristics than sludge ashes without additives were obtained at lower treatment temperatures. PMID:17150348

  20. [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.

  1. A small Tim homohexamer in the relict mitochondrion of Cryptosporidium.

    PubMed

    Alcock, Felicity; Webb, Chaille T; Dolezal, Pavel; Hewitt, Victoria; Shingu-Vasquez, Miguel; Likić, Vladimir A; Traven, Ana; Lithgow, Trevor

    2012-01-01

    The apicomplexan parasite Cryptosporidium parvum possesses a mitosome, a relict mitochondrion with a greatly reduced metabolic capability. This mitosome houses a mitochondrial-type protein import apparatus, but elements of the protein import pathway have been reduced, and even lost, through evolution. The small Tim protein family is a case in point. The genomes of C. parvum and related species of Cryptosporidium each encode just one small Tim protein, CpTimS. This observation challenged the tenet that small Tim proteins are always found in pairs as α3β3 hexamers. We show that the atypical CpTimS exists as a relatively unstable homohexamer, shedding light both on the early evolution of the small Tim protein family and on small Tim hexamer formation in contemporary eukaryotes.

  2. [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

  3. [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

  4. 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

  5. Fates of chlorinated volatile organic compounds in aerobic biological treatment processes: the effects of aeration and sludge addition.

    PubMed

    Chen, Wei-Hsiang; Yang, Wen-Ben; Yuan, Chung-Shin; Yang, Jun-Chen; Zhao, Qing-Liang

    2014-05-01

    The emission of volatile organic compounds (VOCs) from wastewater treatment plants (WWTPs) is becoming an environmental issue of increasing concern. As biological treatment has been considered as one important approach for VOC removal, lab-scale batch experiments were conducted in this study to investigate the fates of four chlorinated hydrocarbons, including chloroform, carbon tetrachloride, trichloroethylene (TCE), and tetrachloroethylene (PERC), in the biological treatment processes with respect to the effects of aeration and sludge addition. The VOC concentrations in the phases of air, water, and sludge under four simulated treatment stages (the first sedimentation, the forepart and rear part of aerobic biological treatment, and the second sedimentation) were analyzed. The results were used to understand the three-phase partitioning of these compounds and to estimate their potentials for volatilization and biological sorption and degradation in these technologies with the concept of fugacity. It was observed that the VOCs were mainly present in the water phase through the experiments. The effects of aeration or sludge addition on the fates of these VOCs occurred but appeared to be relatively limited. The concentration distributions of the VOCs were well below the reported partitioning coefficients. It was suggested that these compounds were unsaturated in the air and sludge phases, enhancing their potentials for volatilization and biological sorption/degradation through the processes. However, the properties of these chlorinated VOCs such as the volatility, polarity, or even biodegradability caused by their structural characteristics (e.g., the number of chlorine, saturated or unsaturated) may represent more significant factors for their fates in the aerobic biological treatment processes. These findings prove the complication behind the current knowledge of VOC pollutions in WWTPs and are of help to manage the adverse impacts on the environment and public

  6. Monoclonal antibody immunotherapy in nude mice persistently infected with Cryptosporidium parvum.

    PubMed Central

    Bjorneby, J M; Hunsaker, B D; Riggs, M W; Perryman, L E

    1991-01-01

    Three groups of congenitally athymic nude mice were persistently infected following oral administration of 2 x 10(7) Cryptosporidium parvum oocysts. Two groups were treated once daily for 10 days with either neutralizing monoclonal antibody (MAb) 17.41 or an isotype control MAb. The third group received no treatment. Intestinal-infection scores were significantly decreased in nude mice treated with MAb 17.41 compared with isotype control MAb-treated and nontreated control groups (P less than 0.005). Biliary and pancreatic cryptosporidial-infection scores were similar for the MAb 17.41-treated and isotype control MAb-treated groups (P greater than 0.05). Images PMID:1997419

  7. 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

  8. 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

  9. Infectious Cryptosporidium parvum oocysts in final reclaimed effluent

    USGS Publications Warehouse

    Gennaccaro, A.L.; McLaughlin, M.R.; Quintero-Betancourt, W.; Huffman, D.E.; Rose, J.B.

    2003-01-01

    Water samples collected throughout several reclamation facilities were analyzed for the presence of infectious Cryptosporidium parvum by the focus detection method-most-probable-number cell culture technique. Results revealed the presence of infectious C. parvum oocysts in 40% of the final disinfected effluent samples. Sampled effluent contained on average seven infectious oocysts per 100 liters. Thus, reclaimed water is not pathogen free but contains infectious C. parvum.

  10. Characterisation of a Cryptosporidium isolate from water buffalo (Bubalus bubalis) by sequencing of a fragment of the Cryptosporidium oocyst wall protein gene (COWP).

    PubMed

    Gómez-Couso, H; Amar, C F L; McLauchlin, J; Ares-Mazás, E

    2005-07-15

    Cryptosporidium oocysts were detected using a direct immunofluorescence antibody test in the faeces of an asymptomatic water buffalo (Bubalus bubalis) heifer from a dairy farm close to Santiago de Compostela (NW Spain). Oocysts were morphologically indistinguishable from Cryptosporidium parvum. Using DNA extracted from this sample and a PCR-RFLP analysis of a 341 base pairs fragment of the Cryptosporidium oocyst wall protein (COWP) gene, a previously undescribed fragment pattern was generated. The COWP gene fragment was cloned and sequencing analyses revealed it to be similar to the C. parvum 'pig' genotype but with four base pairs substitutions.

  11. 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

  12. 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

  13. 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.

  14. 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

  15. 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.

  16. Giardia and Cryptosporidium in water buffaloes (Bubalus bubalis).

    PubMed

    Rinaldi, L; Musella, V; Condoleo, R; Saralli, G; Veneziano, V; Bruni, G; Condoleo, R U; Cringoli, G

    2007-04-01

    A cross-sectional survey of Giardia duodenalis and Cryptosporidium parvum infection in the water buffalo (Bubalus bubalis) was carried out in central Italy. A geographical information system (GIS) was constructed utilizing as data-layers the topographic base map and the digital aerial photographs of the study area, as well as the geo-referenced points of all the buffalo farms. The survey was conducted on a sample of 90 farms, selected using a grid approach followed by proportional allocation. For this purpose, a grid representing quadrants of 5 x 5 km was overlaid on the study area within the GIS. As a result, the study area was divided in equal quadrants, and the number of farms sampled in each quadrant was proportional to the total number of study population in that quadrant. On each farm, faecal samples were collected per rectum from three to five asymptomatic buffalo calves, aged from 1 to 9 weeks. The total number of faecal samples collected was 347. Each faecal sample was tested for the presence of copro-antigens of G. duodenalis and of C. parvum using two commercially available enzyme-linked immunosorbent assays. Out of the 90 farms, 27 (30.0%) resulted positive for G. duodenalis and 22 (24.4%) for C. parvum. Co-infection was found in ten (11.1%) farms. With respect to animals, out of the 347 faecal samples, 63 (18.1%) were found to have antigens of G. duodenalis and 51 (14.7%) of C. parvum. Co-infection was found in ten buffalo calves (2.9%). The results of the logistic regression models showed a positive association between the positivity to G. duodenalis and the presence of sheep on farm and between the positivity to C. parvum and the high number of buffaloes on farms. No significant co-infection between the two protozoa was found. In conclusion, the findings of the present study, derived from a systematic territorial survey planned with GIS, are noteworthy because they provided additional data on C. parvum and the first evidence of G. duodenalis

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

    PubMed Central

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

    2001-01-01

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

  18. 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...

  19. 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...

  20. 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...

  1. 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...

  2. 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...

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

    PubMed

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

    2013-01-01

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

  4. 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...

  5. 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...

  6. 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...

  7. 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...

  8. 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

  9. 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

  10. 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

  11. Cryptosporidium from a free-ranging marsupial host: bandicoots in urban Australia.

    PubMed

    Dowle, Matthew; Hill, Nichola J; Power, Michelle L

    2013-11-15

    Expansion of human settlement has increased the interface between people and bandicoots with implications for the emergence and spread of zoonotic parasites. The host status of bandicoots inhabiting suburban areas and their potential role in Cryptosporidium transmission remains unresolved. Our study aimed to determine the prevalence and identity of Cryptosporidium in two sympatric bandicoot species. Cryptosporidium signatures were detected in twelve bandicoot faecal samples (n=98) through amplification of the 18S rRNA. Phylogenetic inference placed the isolates in a clade with Cryptosporidium parvum, a species with a broad host range and zoonotic potential, or loosely related to Cryptosporidium hominis. However, the identity of the bandicoot isolates was not fully resolved and whether they were infected or simply passively transmitting oocysts is unknown. This study revealed that free-ranging bandicoots of northern Sydney were shedding Cryptosporidium oocysts at a prevalence of 12.2% (95% CI [6.76, 20.8]), similar to marsupial species that act as reservoirs for Cryptosporidium. Our findings expand the range of hosts known to shed Cryptosporidium in urban areas.

  12. 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

  13. 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

  14. Early treatment revisions by addition or switch for type 2 diabetes: impact on glycemic control, diabetic complications, and healthcare costs

    PubMed Central

    Schwab, Phil; Saundankar, Vishal; Bouchard, Jonathan; Wintfeld, Neil; Suehs, Brandon; Moretz, Chad; Allen, Elsie; DeLuzio, Antonio

    2016-01-01

    Background The study examined the prevalence of early treatment revisions after glycosylated hemoglobin (HbA1c) ≥9.0% (75 mmol/mol) and estimated the impact of early treatment revisions on glycemic control, diabetic complications, and costs. Research design and methods A retrospective cohort study of administrative claims data of plan members with type 2 diabetes and HbA1c ≥9.0% (75 mmol/mol) was completed. Treatment revision was identified as treatment addition or switch. Glycemic control was measured as HbA1c during 6–12 months following the first qualifying HbA1c ≥9.0% (75 mmol/mol) laboratory result. Complications severity (via Diabetes Complication Severity Index (DCSI)) and costs were measured after 12, 24, and 36 months. Unadjusted comparisons and multivariable models were used to examine the relationship between early treatment revision (within 90 days of HbA1c) and outcomes after controlling for potentially confounding factors measured during a 12-month baseline period. Results 8463 participants were included with a mean baseline HbA1c of 10.2% (75 mmol/mol). Early treatment revision was associated with greater reduction in HbA1c at 6–12 months (−2.10% vs −1.87%; p<0.001). No significant relationship was observed between early treatment revision and DCSI at 12, 24, or 36 months (p=0.931, p=0.332, and p=0.418). Total costs, medical costs, and pharmacy costs at 12, 24, or 36 months were greater for the early treatment revision group compared with the delayed treatment revision group (all p<0.05). Conclusions The findings suggest that in patients with type 2 diabetes mellitus, treatment revision within 90 days of finding an HbA1c ≥9.0% is associated with a greater level of near-term glycemic control and higher cost. The impact on end points such as diabetic complications may not be realized over relatively short time frames. PMID:26925237

  15. The efficacy of simulated solar disinfection (SODIS) against Ascaris, Giardia, Acanthamoeba, Naegleria, Entamoeba and Cryptosporidium.

    PubMed

    Heaselgrave, Wayne; Kilvington, Simon

    2011-08-01

    The antimicrobial activity of simulated solar disinfection (SODIS) in the presence and absence of riboflavin against various protozoa and helminth organisms was investigated in this study. Assays were conducted in transparent 12 well microtitre plates containing a suspension of test organisms in the presence or absence of 250 μM riboflavin. Plates were exposed to simulated sunlight at an optical irradiance of 550 Wm(-2) (watts per square metre) delivered from a SUNTEST™ CPS+ solar simulator. Aliquots of the test suspensions were taken at set time points and the viability of the test organisms was determined by either culture, microscopy or flow cytometry where applicable. With Acanthamoeba, Naegleria, Entamoeba and Giardia exposure to SODIS at an optical irradiance of 550 Wm(-2) for up to 6h resulted in significant inactivation of these organisms. The addition of riboflavin to this system significantly increased the level of inactivation observed with cysts of A. castellanii. With Cryptosporidium oocysts and Ascaris ova exposure to SODIS in the presence and absence of riboflavin for 6-8h resulted in a negligible reduction in viability of both organisms. In this present study we have been able to show that SODIS is effective against a variety of previously untested waterborne organisms and with A. castellanii cysts the addition of micro-molar concentrations of riboflavin can enhance cyst inactivation. However, care must be taken as Ascaris larvae continue to develop inside the ova after exposure to SODIS and Cryptosporidium remain impermeable to propidium iodide staining indicating they may still be infectious.

  16. Addition of bisphosphonate to antibiotic and anti-inflammatory treatment reduces bone resorption in experimental Staphylococcus aureus-induced arthritis.

    PubMed

    Verdrengh, Margareta; Carlsten, Hans; Ohlsson, Claes; Tarkowski, Andrej

    2007-03-01

    Bacterial arthritis is a disease with high morbidity leading to rapidly progressive bone resorption. We have shown earlier that treatment with antibiotics in combination with corticosteroids decreases joint inflammation and mortality but does not significantly affect bone/cartilage destruction of the joints. This study was performed to assess the effect of treatment with bisphosphonate [zoledronic acid (ZA)] in combination with antibiotics and corticosteroids, on the course and outcome of Staphlococcus aureus-induced arthritis. Three days after intravenous inoculation with S. aureus, mice were treated with antibiotics alone, ZA alone, ZA and antibiotics, or ZA combined with antibiotics and corticosteroids, respectively. One group served as controls and received PBS. Clinical assessment of arthritis was performed as well as histological analysis of bone and cartilage destruction in the joints. One femur from each mouse was collected for bone mineral density (BMD) analysis. In addition, serum levels of type I collagen fragments (RatLaps), and osteocalcin, markers for osteoclastic and osteoblastic activity, respectively, were analyzed. Mice treated with ZA and antibiotics or with ZA in combination with antibiotics and corticosteroids lost significantly less in trabecular bone density compared to infected control mice. Furthermore, the addition of corticosteroids to animals treated with ZA and antibiotics, significantly decreased serum levels of RatLaps and osteocalcin, compared to animals treated with ZA and antibiotics or ZA alone. Treatment with bisphosphonates in combination with antimicrobial agents and corticosteroids significantly decreases the activity of osteoclasts in septic arthritis, thereby reducing the risk of skeletal destruction.

  17. Probiotics in addition to antibiotics for the treatment of acute tonsillitis: a randomized, placebo-controlled study.

    PubMed

    Gilbey, P; Livshits, L; Sharabi-Nov, A; Avraham, Y; Miron, D

    2015-05-01

    Probiotics are live microorganisms which, when administered in adequate amounts, confer a health benefit on the host. The probiotic Streptococcus salivarius has been shown to be effective in reducing the frequency of recurrent pharyngeal infections in children and adult populations. However, probiotics have not yet been evaluated in the treatment of acute pharyngotonsillitis in adults. We aimed to examine whether the addition of S. salivarius probiotics to the routine therapy of acute pharyngotonsillitis in adult patients may shorten disease duration and reduce symptom severity. This study was a prospective, randomized, placebo-controlled, double-blinded study comparing treatment with probiotics to placebo in addition to antibiotics in patients who were hospitalized with severe pharyngotonsillitis. Laboratory results, pain levels, body temperature, and daily volume of fluids consumed were recorded for both groups. Sixty participants were recruited, 30 for each group. No statistically significant differences between the two groups were observed regarding any of the major clinical and laboratory parameters examined. Supplement probiotic treatment with S. salivarius in patients with acute pharyngotonsillitis treated with penicillin is ineffective in relation to the parameters examined in this study and we cannot, therefore, recommend the use of S. salivarius during active pharyngotonsillar infection treated with penicillin.

  18. Changes in the structure and function of microbial communities in drinking water treatment bioreactors upon addition of phosphorus.

    PubMed

    Li, Xu; Upadhyaya, Giridhar; Yuen, Wangki; Brown, Jess; Morgenroth, Eberhard; Raskin, Lutgarde

    2010-11-01

    Phosphorus was added as a nutrient to bench-scale and pilot-scale biologically active carbon (BAC) reactors operated for perchlorate and nitrate removal from contaminated groundwater. The two bioreactors responded similarly to phosphorus addition in terms of microbial community function (i.e., reactor performance), while drastically different responses in microbial community structure were detected. Improvement in reactor performance with respect to perchlorate and nitrate removal started within a few days after phosphorus addition for both reactors. Microbial community structures were evaluated using molecular techniques targeting 16S rRNA genes. Clone library results showed that the relative abundance of perchlorate-reducing bacteria (PRB) Dechloromonas and Azospira in the bench-scale reactor increased from 15.2% and 0.6% to 54.2% and 11.7% after phosphorus addition, respectively. Real-time quantitative PCR (qPCR) experiments revealed that these increases started within a few days after phosphorus addition. In contrast, after phosphorus addition, the relative abundance of Dechloromonas in the pilot-scale reactor decreased from 7.1 to 0.6%, while Zoogloea increased from 17.9 to 52.0%. The results of this study demonstrated that similar operating conditions for bench-scale and pilot-scale reactors resulted in similar contaminant removal performances, despite dramatically different responses from microbial communities. These findings suggest that it is important to evaluate the microbial community compositions inside bioreactors used for drinking water treatment, as they determine the microbial composition in the effluent and impact downstream treatment requirements for drinking water production. This information could be particularly relevant to drinking water safety, if pathogens or disinfectant-resistant bacteria are detected in the bioreactors.

  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. Cryptosporidium spp. from small mammals in the New York City watershed.

    PubMed

    Ziegler, Peter E; Wade, Susan E; Schaaf, Stephanie L; Chang, Yung-Fu; Mohammed, Hussni O

    2007-10-01

    The objective of this study was to assess the potential role that wildlife plays in environmental degradation of watersheds through the contamination of the water supply with zoonotic genotypes of Cryptosporidium. Cryptosporidium isolates recovered from wildlife in the New York City (NYC) watershed were examined to determine genotype using a polymerase chain reaction protocol targeting the 18-Small Subunit (SSU) rRNA locus. Seventy-seven DNA samples recovered from 12 wildlife host species captured in the NYC watershed were amplified and sequenced. Data on risk factors associated with the perpetuation of these genotypes also were collected and analyzed. Although many genotypes appeared to be host-specific, 38% of the samples examined were identified as Cryptosporidium parvum, indicating the presence of zoonotic Cryptosporidium. Adult animals were more likely to shed the zoonotic strains of Cryptosporidium spp. Animals captured in the fall and winter were more likely to be infected with C. parvum than those captured in spring and summer.

  1. Molecular Characterization of Cryptosporidium Species in Children with Diarrhea in North West of Iran

    PubMed Central

    Mahdavi Poor, Behroz; Rashedi, Jalil; Asgharzadeh, Mohammad; Fallah, Esmaeel; Hatam-Nahavandi, Kareem; Dalimi, Abdolhossein

    2015-01-01

    Cryptosporidium is one of the most common causes of childhood diarrhea in developing countries. The aim of this randomized pilot study was to detect and characterize infective species and determine the genotypes of Cryptosporidium parasites in pediatric patients suffering from diarrhea in North West of Iran. A total of 113 fecal samples were collected from diarrheic children hospitalized in Tabriz Pediatric Hospital. The amplification of small subunit ribosomal RNA gene was performed using a nested polymerase chain reaction protocol and its products were digested using two restriction enzymes for Cryptosporidium species and genotype differentiation. Cryptosporidium oocysts were found in 2 (1.76%) children with diarrhea and restriction pattern revealed the presence of C.parvum bovine genotype in both positive fecal samples. The findings indicate that Cryptosporidium parvum is responsible for cryptosporidiosis in children in the study region and probably zoonotic transmission is the predominant route of parasite transmission. PMID:27014648

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

    PubMed

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

    2016-02-01

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

  3. Molecular Characterization of Cryptosporidium Species in Children with Diarrhea in North West of Iran.

    PubMed

    Mahdavi Poor, Behroz; Rashedi, Jalil; Asgharzadeh, Mohammad; Fallah, Esmaeel; Hatam-Nahavandi, Kareem; Dalimi, Abdolhossein

    2015-01-01

    Cryptosporidium is one of the most common causes of childhood diarrhea in developing countries. The aim of this randomized pilot study was to detect and characterize infective species and determine the genotypes of Cryptosporidium parasites in pediatric patients suffering from diarrhea in North West of Iran. A total of 113 fecal samples were collected from diarrheic children hospitalized in Tabriz Pediatric Hospital. The amplification of small subunit ribosomal RNA gene was performed using a nested polymerase chain reaction protocol and its products were digested using two restriction enzymes for Cryptosporidium species and genotype differentiation. Cryptosporidium oocysts were found in 2 (1.76%) children with diarrhea and restriction pattern revealed the presence of C.parvum bovine genotype in both positive fecal samples. The findings indicate that Cryptosporidium parvum is responsible for cryptosporidiosis in children in the study region and probably zoonotic transmission is the predominant route of parasite transmission. PMID:27014648

  4. Treatment of a simulated textile wastewater in a sequencing batch reactor (SBR) with addition of a low-cost adsorbent.

    PubMed

    Santos, Sílvia C R; Boaventura, Rui A R

    2015-06-30

    Color removal from textile wastewaters, at a low-cost and consistent technology, is even today a challenge. Simultaneous biological treatment and adsorption is a known alternative to the treatment of wastewaters containing biodegradable and non-biodegradable contaminants. The present work aims at evaluating the treatability of a simulated textile wastewater by simultaneously combining biological treatment and adsorption in a SBR (sequencing batch reactor), but using a low-cost adsorbent, instead of a commercial one. The selected adsorbent was a metal hydroxide sludge (WS) from an electroplating industry. Direct Blue 85 dye (DB) was used in the preparation of the synthetic wastewater. Firstly, adsorption kinetics and equilibrium were studied, in respect to many factors (temperature, pH, WS dosage and presence of salts and dyeing auxiliary chemicals in the aqueous media). At 25 °C and pH 4, 7 and 10, maximum DB adsorption capacities in aqueous solution were 600, 339 and 98.7 mg/g, respectively. These values are quite considerable, compared to other reported in literature, but proved to be significantly reduced by the presence of dyeing auxiliary chemicals in the wastewater. The simulated textile wastewater treatment in SBR led to BOD5 removals of 53-79%, but color removal was rather limited (10-18%). The performance was significantly enhanced by the addition of WS, with BOD5 removals above 91% and average color removals of 60-69%.

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

    PubMed

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

    2015-08-01

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

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

    PubMed Central

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

    2012-01-01

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

  7. Spatial and temporal distribution of Cryptosporidium and Giardia in a drinking water resource: implications for monitoring and risk assessment.

    PubMed

    Burnet, Jean-Baptiste; Penny, Christian; Ogorzaly, Leslie; Cauchie, Henry-Michel

    2014-02-15

    Because of their significant public health impact, waterborne Cryptosporidium and Giardia have been monitored in surface water in order to assess microbial quality of water bodies used for drinking water production and/or for recreational purposes. In this context, sampling strategy is of key importance and should be representative enough to appropriately assess the related microbial risk. This, however, requires sound knowledge on the behaviour of both pathogens in water. In the present study, the spatial and temporal distribution of Cryptosporidium and Giardia was explored in the rural Upper-Sûre watershed used for drinking water production in Luxembourg. By subdividing it into three compartments including (i) sub-catchments, (ii) the Sûre River fed by the sub-catchments and (iii) the Upper-Sûre reservoir fed by the Sûre River, parasite distribution was assessed using sampling designs adapted to the hydro-dynamic characteristics of the respective compartments. Results highlighted the high spatial and temporal variability in parasite distribution at watershed scale, as well as the prevalence of Giardia over Cryptosporidium. Besides land use features and catchment characteristics, hydro-climatology appeared to be a major driver of parasite behaviour in the watershed. It introduced a seasonal trend in their occurrence, highest densities being detected during the wet season. Peaks of contamination triggered out by rainfall-induced runoff were further observed in the three compartments. In the Sûre River, Cryptosporidium and Giardia fluxes peaked at 10(9) and 10(10) (oo)cysts.d(-1), respectively, and were discharged into the drinking water reservoir, where they underwent a 2 to 3 log10 removal rate. Despite this, parasite fluxes entering the drinking water treatment plant were still high (10(6) to 10(7) (oo)cysts.d(-1)) and stressed on the need for improved watershed management upstream the water treatment barrier. The catchment-wide analysis described here

  8. 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

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

    PubMed

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

    2012-05-01

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

  10. 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. 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

  12. 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

  13. 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.

  14. First description of heterogeneity in 18S rRNA genes in the haploid genome of Cryptosporidium andersoni Kawatabi type.

    PubMed

    Ikarashi, Makoto; Fukuda, Yasuhiro; Honma, Hajime; Kasai, Kenji; Kaneta, Yoshiyasu; Nakai, Yutaka

    2013-09-01

    The Apicomplexan Cryptosporidium andersoni, is a species of gastric Cryptosporidium, is frequently detected in older calves and adult cattle. Genotyping analyses based on 18S ribosomal RNA gene sequences have been performed on a novel C. andersoni genotype, namely the Kawatabi type, and the oocysts were classified into two distinct groups genotypically: Type A (the sequence in GenBank) and Type B (with a thymine nucleotide insertion not in Type A). This study analyzed 3775 cattle at a slaughterhouse and 310 cattle at a farm using microscopy and found 175 Cryptosporidium-positive animals: 171 from the slaughterhouse and four from the farm, and all infecting parasites were determined to be C. andersoni from 18S rRNA gene sequences determined from fecal DNA. In genotyping analyses with single isolated oocysts, about a half of analyzed ones were clearly classified into well known two genotypes (Type A and B). In addition to these two known genotypes, we have detected some oocysts showing mixed signals of Types A and B in the electropherogram from the automated sequencer (the Type C genotype). To determine the genotypic composition of sporozoites carried by the Type C oocysts, we analyzed their 18S rRNA gene sequences using a single sporozoite isolation procedure. Some sporozoites were classified as either Type A or Type B. However, more than half of the analyzed isolated sporozoites showed a mixed signal identical to that of Type C oocysts, and both the Type A and B signals were surely detectable from such sporozoites after a cloning procedure. In conclusion, C. andersoni carries two different genotypes heterogeneously in its haploid genome.

  15. 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

  16. Survey and genetic characterization of wastewater in Tunisia for Cryptosporidium spp., Giardia duodenalis, Enterocytozoon bieneusi, Cyclospora cayetanensis and Eimeria spp.

    PubMed

    Ben Ayed, Layla; Yang, Wenli; Widmer, Giovanni; Cama, Vitaliano; Ortega, Ynes; Xiao, Lihua

    2012-09-01

    The microbial diversity of wastewater used for irrigation and fertilization was assessed using specific polymerase chain reaction (PCR) assays to detect and genotype several pathogenic protists including Cryptosporidium spp., Giardia duodenalis, Cyclospora spp., Eimeria spp. and Enterocytozoon bieneusi. A total of 220 wastewater samples (110 raw, 110 treated) and 12 sludge samples were collected from 2005 to 2008 from 18 treatment plants located throughout Tunisia. Except for Cyclospora, which was detected only once, E. bieneusi (61%), G. duodenalis (28%), Cryptosporidium spp. (27%) and Eimeria spp. (45%) were frequently observed in wastewater and sludge. Sequencing of PCR products showed that C. hominis, C. andersoni, G. duodenalis sub-assemblage A-II and E. bieneusi genotypes D and IV were the most prevalent. An analysis of the distribution of 209 internal transcribed spacer sequences of E. bieneusi originating from wastewater at the 18 treatment plants showed a similar genetic diversity, regardless of the geographical location. The identification of these parasite species and genotypes and of host-specific Eimeria species indicates that the microbial quality of wastewater was impacted by humans, livestock and rodents. Given the public health risks that some of these parasites represent, guidelines on wastewater usage are needed to minimize human exposure to these pathogens.

  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.

  18. A comparison of additional treatment processes to limit particle accumulation and microbial growth during drinking water distribution.

    PubMed

    Liu, G; Lut, M C; Verberk, J Q J C; Van Dijk, J C

    2013-05-15

    Water quality changes, particle accumulation and microbial growth occurring in pilot-scale water distribution systems fed with normally treated and additional treated groundwater were monitored over a period of almost one year. The treatment processes were ranked in the following order: nanofiltration (NF) > (better than) ultrafiltration (UF) > ion exchange (IEX) for limiting particle accumulation. A different order was found for limiting overall microbial growth: NF > IEX > UF. There were strong correlations between particle load and particle accumulation, and between nutrient load and microbial growth. It was concluded that particle accumulation can be controlled by reducing the particle load in water treatment plants; and the microbial growth can be better controlled by limiting organic nutrients rather than removing biomass in water treatment plants. The major focus of this study was on microbial growth. The results demonstrated that growth occurred in all types of treated water, including the phases of bulk water, biofilm and loose deposits. Considering the growth in different phases, similar growth in bulk water was observed for all treatments; NF strongly reduced growth both in loose deposits and in biofilm; UF promoted growth in biofilm, while strongly limiting growth in loose deposits. IEX had good efficiency in between UF and NF, limiting both growths in loose deposits and in biofilm. Significant growth was found in loose deposits, suggesting that loose deposit biomass should be taken into account for growth evaluation and/or prediction. Strong correlations were found between microbial growth and pressure drop in a membrane fouling simulator which proved that a membrane fouling simulator can be a fast growth predictor (within a week). Different results obtained by adenosine triphosphate and flow cytometry cell counts revealed that ATP can accurately describe both suspended and particle-associated biomass, and flow cytometry files of TCC measurements needs

  19. 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

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

    PubMed Central

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

    2005-01-01

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

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

    PubMed

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

    2005-10-01

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

  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. 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.

  6. Randomized clinical trial assessing whether additional massage treatments for chronic neck pain improve 12- and 26-week outcomes

    PubMed Central

    Cook, Andrea J.; Wellman, Robert D.; Cherkin, Daniel C.; Kahn, Janet R.; Sherman, Karen J.

    2015-01-01

    Background Context This is the first study to systematically evaluate the value of a longer treatment period for massage. We provide a framework of how to conceptualize an optimal dose in this challenging setting of non-pharmacological treatments. Purpose To determine the optimal dose of massage for neck pain. Study Design/Setting Two-phase randomized trial for persons with chronic non-specific neck pain. Primary randomization to one of 5 groups receiving 4 weeks of massage (30 minutes 2×/ or 3×/week or 60 minutes 1×, 2×, or 3×/week). Booster randomization of participants to receive an additional 6 massages, 60 minute 1×/week, or no additional massage. Patient Sample 179 participants from Group Health and the general population of Seattle, WA USA recruited between June 2010 and August 2011. Outcome Measures Primary outcomes self-reported neck-related dysfunction (Neck Disability Index) and pain (0–10 scale) were assessed at baseline, 12, and 26 weeks. Clinically meaningful improvement was defined as >5 point decrease in dysfunction and > 30% decrease in pain from baseline. Methods Clinically meaningful improvement for each primary outcome with both follow-up times was analyzed using adjusted modified Poisson generalized estimating equations. Secondary analyses for the continuous outcomes used linear generalized estimating equations. This study was funded the National Center for Complementary and Alternative Medicine, NIH, USA (R01 AT004411). The funders had no role in the interpretation or reporting of results. Results There were no observed differences by primary treatment group at 12 or 26 weeks. Those receiving booster dose had improvements in both dysfunction and pain at 12 weeks (dysfunction: RR=1.56(1.08–2.25), P=0.018; pain: RR=1.25(0.98–1.61); P=0.077), but those were non-significant at 26 weeks (dysfunction: RR=1.22(0.85–1.74); pain: RR=1.09(0.82–1.43)). Subgroup analysis by primary and booster treatments found the booster dose only

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

    PubMed

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

    2013-01-01

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

  8. 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

  9. Prevalence of Cryptosporidium infection in calves in France.

    PubMed

    Lefay, D; Naciri, M; Poirier, P; Chermette, R

    2000-03-28

    Two multicentre surveys were conducted in France to estimate the prevalence of Cryptosporidium infection in calves using qualitative ELISA for detection of Cryptosporidium coproantigens and oocysts. The first survey involved 4-12-day-old calves in six dairy-calf distribution centres, collecting calves from seven Administrative Regions (Aquitaine, Bretagne, Franche-Comté, Lorraine, Normandie, Nord, Pays de Loire). For each region, 20 calves were selected every month for 12 consecutive months (October 1995-September 1996). Prevalence of Cryptosporidium infection was 17.9% (Confidence Intervals (C.I.) 95%=[16.1%; 19.8%]) among the 1628 selected calves, of which only 5.3% had diarrhoea. The second survey conducted between November 1995 and May 1996 involved 4-21-day-old calves examined by veterinary practitioners who selected 189 livestock farms of dairy- or suckler-type in ten Administrative Departments (Allier, Cantal, Creuse, Doubs, Ille-et-Vilaine, Maine-et-Loire, Manche, Pas-de-Calais, Saône-et-Loire, Vendée). Cryptosporidia were detected in 105 (55.6%) of the farms. Among the 440 calves examined, of which 398 (90.5%) presented diarrhoea, cryptosporidia were found in 191 animals, i.e. a prevalence of 43.4% (C.I. 95%=[38. 8%; 48.0%]). Breed of calves and type of housing had very little impact on prevalence in this survey. Some regional variations could be noticed, even if cryptosporidia infection is widespread. Monthly variations could be related to seasonal peaks in calving with a lower infection rate during summer.

  10. Isolation and enrichment of Cryptosporidium DNA and verification of DNA purity for whole-genome sequencing.

    PubMed

    Guo, Yaqiong; Li, Na; Lysén, Colleen; Frace, Michael; Tang, Kevin; Sammons, Scott; Roellig, Dawn M; Feng, Yaoyu; Xiao, Lihua

    2015-02-01

    Whole-genome sequencing of Cryptosporidium spp. is hampered by difficulties in obtaining sufficient, highly pure genomic DNA from clinical specimens. In this study, we developed procedures for the isolation and enrichment of Cryptosporidium genomic DNA from fecal specimens and verification of DNA purity for whole-genome sequencing. The isolation and enrichment of genomic DNA were achieved by a combination of three oocyst purification steps and whole-genome amplification (WGA) of DNA from purified oocysts. Quantitative PCR (qPCR) analysis of WGA products was used as an initial quality assessment of amplified genomic DNA. The purity of WGA products was assessed by Sanger sequencing of cloned products. Next-generation sequencing tools were used in final evaluations of genome coverage and of the extent of contamination. Altogether, 24 fecal specimens of Cryptosporidium parvum, C. hominis, C. andersoni, C. ubiquitum, C. tyzzeri, and Cryptosporidium chipmunk genotype I were processed with the procedures. As expected, WGA products with low (<16.0) threshold cycle (CT) values yielded mostly Cryptosporidium sequences in Sanger sequencing. The cloning-sequencing analysis, however, showed significant contamination in 5 WGA products (proportion of positive colonies derived from Cryptosporidium genomic DNA, ≤25%). Following this strategy, 20 WGA products from six Cryptosporidium species or genotypes with low (mostly <14.0) CT values were submitted to whole-genome sequencing, generating sequence data covering 94.5% to 99.7% of Cryptosporidium genomes, with mostly minor contamination from bacterial, fungal, and host DNA. These results suggest that the described strategy can be used effectively for the isolation and enrichment of Cryptosporidium DNA from fecal specimens for whole-genome sequencing.

  11. 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

  12. A waterborne outbreak and detection of cryptosporidium oocysts in drinking water of an older high-rise apartment complex in seoul.

    PubMed

    Cho, Eun-Joo; Yang, Jin-Young; Lee, Eun-Sook; Kim, Se-Chul; Cha, So-Yang; Kim, Sung-Tek; Lee, Man-Ho; Han, Sun-Hee; Park, Young-Sang

    2013-08-01

    From May to June 2012, a waterborne outbreak of 124 cases of cryptosporidiosis occurred in the plumbing systems of an older high-rise apartment complex in Seoul, Republic of Korea. The residents of this apartment complex had symptoms of watery diarrhea and vomiting. Tap water samples in the apartment complex and its adjacent buildings were collected and tested for 57 parameters under the Korean Drinking Water Standards and for additional 11 microbiological parameters. The microbiological parameters included total colony counts, Clostridium perfringens, Enterococcus, fecal streptococcus, Salmonella, Shigella, Pseudomonas aeruginosa, Cryptosporidium oocysts, Giardia cysts, total culturable viruses, and Norovirus. While the tap water samples of the adjacent buildings complied with the Korean Drinking Water Standards for all parameters, fecal bacteria and Cryptosporidium oocysts were detected in the tap water samples of the outbreak apartment complex. It turned out that the agent of the disease was Cryptosporidium parvum. The drinking water was polluted with sewage from a septic tank in the apartment complex. To remove C. parvum oocysts, we conducted physical processes of cleaning the water storage tanks, flushing the indoor pipes, and replacing old pipes with new ones. Finally we restored the clean drinking water to the apartment complex after identification of no oocysts.

  13. 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

  14. Addition of immunosuppressive treatment to hemoperfusion is associated with improved survival after paraquat poisoning: a nationwide study.

    PubMed

    Wu, Wen-Pyng; Lai, Ming-Nan; Lin, Ching-Heng; Li, Yu-Fen; Lin, Ching-Yuang; Wu, Ming-Ju

    2014-01-01

    Paraquat poisoning associates very high mortality rate. Early treatment with hemoperfusion is strongly suggested by animal and human studies. Although the survival benefit of additional immunosuppressive treatment (IST) in combination with hemoperfusion is also reported since 1971, the large-scale randomized control trials to confirm the effects of IST is difficult to be executed. Therefore, we designed this nationwide large-scale population-based retrospective cohort study to investigate the outcome of paraquat poisoning with hemoperfusion and the additional effects of IST combined with hemoperfusion. This nationwide retrospective cohort study utilized data retrieved from the National Health Insurance Research Database (NHIRD) of Taiwan. A total of 1811 hospitalized patients with a diagnosis of paraquat poisoning who received hemoperfusion between 1997 and 2009 were enrolled. The mean age of all 1811 study subjects was 47.3 years. 70% was male. The overall survival rate was only 26.4%. Respiratory failure and renal failure were diagnosed in 56.2% and 36% patients. The average frequency of hemoperfusion was twice. IST was added in 42.2% patients. IST significantly increases survival rate (from 24.3% to 29.3%, P<0.001). The combined IST with methylprednisolone, cyclophosphamide and dexamethasone associates with the highest survival rate (48%, P<0.001). Moreover, patients younger than 45 years of age in the IST group had the best survival (41.0% vs. 33.7%, p<0.001). Our results support the use of IST with hemoperfusion for paraquat-poisoned patients. The best survival effect of IST is the combination of methylprednisolone, cyclophosphamide and daily dexamethasone, especially in patients with younger age.

  15. 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.

  16. 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.

  17. Cryptosporidium in small water systems in Puerto Rico: a pilot study.

    PubMed

    Robinson, Guy; Minnigh, Harvey A; Hunter, Paul R; Chalmers, Rachel M; Ramírez Toro, Graciela I

    2015-09-01

    A pilot study was undertaken to investigate the occurrence of Cryptosporidium in four very small drinking water systems supplying communities in rural Puerto Rico. Water samples (40 L) were collected and oocysts were concentrated by calcium carbonate flocculation, recovered by immunomagnetic separation and detected by immunofluorescence microscopy. Cryptosporidium oocysts were identified in all four systems. This is the first report of evidence of the potential public health risk from this chlorine-resistant pathogen in Puerto Rican small water systems. Further work is warranted to fully assess the health risks that Cryptosporidium and other protozoa pose to populations served by community-managed small drinking water systems. PMID:26322771

  18. Cryptosporidium in small water systems in Puerto Rico: a pilot study.

    PubMed

    Robinson, Guy; Minnigh, Harvey A; Hunter, Paul R; Chalmers, Rachel M; Ramírez Toro, Graciela I

    2015-09-01

    A pilot study was undertaken to investigate the occurrence of Cryptosporidium in four very small drinking water systems supplying communities in rural Puerto Rico. Water samples (40 L) were collected and oocysts were concentrated by calcium carbonate flocculation, recovered by immunomagnetic separation and detected by immunofluorescence microscopy. Cryptosporidium oocysts were identified in all four systems. This is the first report of evidence of the potential public health risk from this chlorine-resistant pathogen in Puerto Rican small water systems. Further work is warranted to fully assess the health risks that Cryptosporidium and other protozoa pose to populations served by community-managed small drinking water systems.

  19. Reduction of salt in pork sausages by the addition of carrot fibre or potato starch and high pressure treatment.

    PubMed

    Grossi, Alberto; Søltoft-Jensen, Jakob; Knudsen, Jes Christian; Christensen, Mette; Orlien, Vibeke

    2012-12-01

    The combined effect of high pressure processing (HPP) (400, 600 and 800 MPa) and carrot fibre (CF) and potato starch (PS) on low salt (1.2%) pork sausages was investigated and compared with high (1.8%) salt sausages. Sausages had a marked increase in whitening with increasing content of fibre or starch, pressure level, and process temperature. The degree of redness was mainly affected by pressure level and heat treatment. An important finding regarding salt reduction was that the use of starch or fibre had more impact on textural properties than the level of salt since Young's modulus and strain at fracture were mainly affected by formulation and HPP. Water binding capacity of low salt sausages was improved to the same level as high salt sausages with HPP and addition of CF or PS particularly by the addition of PS which produced sausages with better sensory properties than CF. The sensory analysis showed that this approach is promising for producing low salt sausages. PMID:22682686

  20. 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

  1. 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.

  2. 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

  3. 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.

  4. Longitudinal Poisson regression to evaluate the epidemiology of Cryptosporidium, Giardia, and fecal indicator bacteria in coastal California wetlands.

    PubMed

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

    2012-05-01

    Fecal pathogen contamination of watersheds worldwide is increasingly recognized, and natural wetlands may have an important role in mitigating fecal pathogen pollution flowing downstream. Given that waterborne protozoa, such as Cryptosporidium and Giardia, are transported within surface waters, this study evaluated associations between fecal protozoa and various wetland-specific and environmental risk factors. This study focused on three distinct coastal California wetlands: (i) a tidally influenced slough bordered by urban and agricultural areas, (ii) a seasonal wetland adjacent to a dairy, and (iii) a constructed wetland that receives agricultural runoff. Wetland type, seasonality, rainfall, and various water quality parameters were evaluated using longitudinal Poisson regression to model effects on concentrations of protozoa and indicator bacteria (Escherichia coli and total coliform). Among wetland types, the dairy wetland exhibited the highest protozoal and bacterial concentrations, and despite significant reductions in microbe concentrations, the wetland could still be seen to influence water quality in the downstream tidal wetland. Additionally, recent rainfall events were associated with higher protozoal and bacterial counts in wetland water samples across all wetland types. Notably, detection of E. coli concentrations greater than a 400 most probable number (MPN) per 100 ml was associated with higher Cryptosporidium oocyst and Giardia cyst concentrations. These findings show that natural wetlands draining agricultural and livestock operation runoff into human-utilized waterways should be considered potential sources of pathogens and that wetlands can be instrumental in reducing pathogen loads to downstream waters.

  5. Longitudinal Poisson Regression To Evaluate the Epidemiology of Cryptosporidium, Giardia, and Fecal Indicator Bacteria in Coastal California Wetlands

    PubMed Central

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

    2012-01-01

    Fecal pathogen contamination of watersheds worldwide is increasingly recognized, and natural wetlands may have an important role in mitigating fecal pathogen pollution flowing downstream. Given that waterborne protozoa, such as Cryptosporidium and Giardia, are transported within surface waters, this study evaluated associations between fecal protozoa and various wetland-specific and environmental risk factors. This study focused on three distinct coastal California wetlands: (i) a tidally influenced slough bordered by urban and agricultural areas, (ii) a seasonal wetland adjacent to a dairy, and (iii) a constructed wetland that receives agricultural runoff. Wetland type, seasonality, rainfall, and various water quality parameters were evaluated using longitudinal Poisson regression to model effects on concentrations of protozoa and indicator bacteria (Escherichia coli and total coliform). Among wetland types, the dairy wetland exhibited the highest protozoal and bacterial concentrations, and despite significant reductions in microbe concentrations, the wetland could still be seen to influence water quality in the downstream tidal wetland. Additionally, recent rainfall events were associated with higher protozoal and bacterial counts in wetland water samples across all wetland types. Notably, detection of E. coli concentrations greater than a 400 most probable number (MPN) per 100 ml was associated with higher Cryptosporidium oocyst and Giardia cyst concentrations. These findings show that natural wetlands draining agricultural and livestock operation runoff into human-utilized waterways should be considered potential sources of pathogens and that wetlands can be instrumental in reducing pathogen loads to downstream waters. PMID:22427504

  6. Differentiation between Viable and Dead Cryptosporidium Oocysts Using Fluorochrome Staining.

    PubMed

    Tomonaga, Tatsuya; Rai, Shiba Kumar; Uga, Shoji

    2016-01-01

    The use of nucleic acid staining with a fluorochrome dye to differentiate viable and dead (heat-killed) Cryptosporidium oocysts was assessed. The specificities (percentage of unstained viable oocysts) and sensitivities (percentage of stained dead oocysts) of the seven tested dyes (SYTO-17® and SYTO-59® to 64®) ranged from 65 to 76% (average 71%) and 83 to 95% (average 91%), respectively. SYTO-59 and SYTO-17 imparted greater color (4+) intensity than the other dyes (2+ or less). Of these two dyes, SYTO-17 exhibited more brightness and slower discoloration and was selected for use in further experiments. The optimum staining time for SYTO-17 at 37℃ was one hour or more (sensitivity of 96%). Dye concentrations of 20 and 30 µM resulted in maximal color intensity, and no further improvement was observed with further increases in dye concentration. Staining a mixture of viable and dead oocysts (1:1 ratio) with 20 µM dye at 37℃ for one hour yielded the expected results (approximately 50%), but no remarkable increase in the percent staining with time (up to 8 hours) was observed. In this study, no ghost oocysts were observed. The present study indicated that the fluorogenic nucleic acid dye SYTO-17 could be used to discriminate between live and dead Cryptosporidium oocysts. PMID:27363397

  7. 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.

  8. 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

  9. 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

  10. Cryptosporidium fayeri n. sp. (Apicomplexa: Cryptosporidiidae) from the Red Kangaroo (Macropus rufus).

    PubMed

    Ryan, Una M; Power, Michelle; Xiao, Lihua

    2008-01-01

    The morphology and infectivity of the oocysts of a new species of Cryptosporidium from the faeces of the red kangaroo (Macropus rufus) are described. Oocysts are structurally indistinguishable from those of Cryptosporidium parvum. Oocysts of the new species are passed fully sporulated, lack sporocysts, and measure 4.5-5.1 microm (mean=4.9) x 3.8-5.0 microm (mean=4.3 microm) with a length to width ratio 1.02:1.18 (mean 1.14) (n=50). Oocysts were not infectious for neonate ARC Swiss mice. Multi-locus analysis of numerous unlinked loci demonstrated this species to be distinct (90.64%-97.88% similarity) from C. parvum. Based on biological and molecular data, this Cryptosporidium infecting marsupials is proposed to be a new species Cryptosporidium fayeri n. sp.

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

    PubMed

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

    2005-01-01

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

  12. 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.

  13. 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).

  14. 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,...

  15. 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, ...

  16. 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...

  17. 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...

  18. 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...

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

    EPA Science Inventory

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

  20. 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...

  1. 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.

  2. 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

  3. 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 ...

  4. 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...

  5. 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

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

    PubMed

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

    1998-12-01

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

  7. 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.

  8. 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...

  9. 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...

  10. 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

  11. CHARACTERIZATION OF CRYPTOSPORIDIUM PARVUM BY MATRIX-ASSISTED LASER DESORPTION -- IONIZATION TIME OF FLIGHT MASS SPECTROMETRY

    EPA Science Inventory

    Matrix assisted laser desorption/ionization (MALDI) mass spectrometry was used to investigate whole and freeze thawed Cryptosporidium parvum oocysts. Whole oocysts revealed some mass spectral features. Reproducible patterns of spectral markers and increased sensitivity were obtai...

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

    PubMed

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

    2005-12-01

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

  13. 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.

  14. 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.

  15. 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

  16. 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

  17. Identification of novel and zoonotic Cryptosporidium species in fish from Papua New Guinea.

    PubMed

    Koinari, M; Karl, S; Ng-Hublin, J; Lymbery, A J; Ryan, U M

    2013-11-15

    There is still limited information on the distribution of Cryptosporidium species and genotypes in fish. The present study investigated the prevalence of Cryptosporidium species in cultured freshwater (n=132), wild freshwater (n=206) and wild marine (n=276) fish in Papua New Guinea (PNG) by PCR screening at the 18S rRNA locus. A total of seven fish (2 cultured freshwater, 1 wild freshwater and 4 wild marine fish) were identified as positive for Cryptosporidium. Specifically, Cryptosporidium was found in four different host species (Nile tilapia, Oreochromis niloticus; silver barb, Puntius gonionotus; mackerel scad, Decapterus maracellus and oblong silver biddy, Gerres oblongus), giving an overall prevalence of 1.14% (95% CI: 0.3-2%, n=7/614). Of the seven positive isolates, five were identified as C. parvum and two were a novel piscine genotype, which we have named piscine genotype 8. Piscine genotype 8 was identified in two marine oblong silver biddies and exhibited 4.3% genetic distance from piscine genotype 3 at the 18S locus. Further subtyping of C. parvum isolates at the 60 kDa glycoprotein (gp60) locus identified 3 C. parvum subtypes (IIaA14G2R1, IIaA15G2R1 and IIaA19G4R1) all of which are zoonotic and a C. hominis subtype (IdA15G1). The zoonotic Cryptosporidium were identified in fish samples from all three groups; cultured and wild freshwater and wild marine fish. Detection of Cryptosporidium among aquaculture fingerlings warrants further research to gain a better understanding of the epidemiology of Cryptosporidium infection in cultured fish. The identification of zoonotic Cryptosporidium genotypes in fish from PNG has important public health implications and should be investigated further.

  18. 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.

  19. 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.

  20. 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.