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Sample records for inactivate cryptosporidium oocysts

  1. Inactivation of oocysts of Cryptosporidium parvum by ultraviolet irradiation

    International Nuclear Information System (INIS)

    Campbell, A.T.; Robertson, L.J.; Snowball, M.R.; Smith, H.V.

    1995-01-01

    Inactivation of oocysts of Cryptosporidium parvum in clean water using a novel design of an ultraviolet disinfection system was assessed by a vital dye assay and by in vitro excystation. The disinfection unit system is designed to expose the oocysts to ultraviolet radiation on two filters, providing a maximum total exposure to ultraviolet radiation of 8748 mW s cm −2 . Results revealed a reduction in oocyst viability of over two logs, indicating that this treatment has exciting potential as an additional treatment for water already treated by conventional methods. However, these data are only preliminary results using one isolate of oocysts and further trials must be conducted before this system could be recommended for use

  2. Batch solar disinfection inactivates oocysts of Cryptosporidium parvum and cysts of Giardia muris in drinking water.

    Science.gov (United States)

    McGuigan, K G; Méndez-Hermida, F; Castro-Hermida, J A; Ares-Mazás, E; Kehoe, S C; Boyle, M; Sichel, C; Fernández-Ibáñez, P; Meyer, B P; Ramalingham, S; Meyer, E A

    2006-08-01

    To determine whether batch solar disinfection (SODIS) can be used to inactivate oocysts of Cryptosporidium parvum and cysts of Giardia muris in experimentally contaminated water. Suspensions of oocysts and cysts were exposed to simulated global solar irradiation of 830 W m(-2) for different exposure times at a constant temperature of 40 degrees C. Infectivity tests were carried out using CD-1 suckling mice in the Cryptosporidium experiments and newly weaned CD-1 mice in the Giardia experiments. Exposure times of > or =10 h (total optical dose c. 30 kJ) rendered C. parvum oocysts noninfective. Giardia muris cysts were rendered completely noninfective within 4 h (total optical dose >12 kJ). Scanning electron microscopy and viability (4',6-diamidino-2-phenylindole/propidium iodide fluorogenic dyes and excystation) studies on oocysts of C. parvum suggest that inactivation is caused by damage to the oocyst wall. Results show that cysts of G. muris and oocysts of C. parvum are rendered completely noninfective after batch SODIS exposures of 4 and 10 h (respectively) and is also likely to be effective against waterborne cysts of Giardia lamblia. These results demonstrate that SODIS is an appropriate household water treatment technology for use as an emergency intervention in aftermath of natural or man-made disasters against not only bacterial but also protozoan pathogens.

  3. Modeling Cryptosporidium spp. Oocyst Inactivation in Bubble-Diffuser Ozone Contactors

    Science.gov (United States)

    1998-07-01

    requirements for Giardia lamblia (G. lamblia) and viruses under the Surface Water Treatment Rule (SWTR). Minimum CT requirements include relatively...parvum and C. muris ) oocysts in ozone bubble-diffuser contactors. The model is calibrated with semi-batch kinetic data, verified with pilot-scale

  4. An Immunoglobulin G1 Monoclonal Antibody Highly Specific to the Wall of Cryptosporidium Oocysts

    Science.gov (United States)

    Weir, C.; Vesey, G.; Slade, M.; Ferrari, B.; Veal, D. A.; Williams, K.

    2000-01-01

    The detection of Cryptosporidium oocysts in drinking water is critically dependent on the quality of immunofluorescent reagents. Experiments were performed to develop a method for producing highly specific antibodies to Cryptosporidium oocysts that can be used for water testing. BALB/c mice were immunized with six different antigen preparations and monitored for immunoglobulin G (IgG) and IgM responses to the surface of Cryptosporidium oocysts. One group of mice received purified oocyst walls, a second group received a soluble protein preparation extracted from the outside of the oocyst wall, and the third group received whole inactivated oocysts. Three additional groups were immunized with sequentially prepared oocyst extracts to provide for a comparison of the immune response. Mice injected with the soluble protein extract demonstrated an IgG response to oocysts surface that was not seen in the whole-oocyst group. Mice injected with whole oocysts showed an IgM response only, while mice injected with purified oocyst walls showed little increase in IgM or IgG levels. Of the additional reported preparations only one, BME (2-mercaptoethanol treated), produced a weak IgM response to the oocyst wall. A mouse from the soluble oocyst extract group yielding a high IgG response was utilized to produce a highly specific IgG1 monoclonal antibody (Cry104) specific to the oocyst surface. Comparative flow cytometric analysis indicated that Cry104 has a higher avidity and specificity to oocysts in water concentrates than other commercially available antibodies. PMID:10973448

  5. Molecular fingerprinting of Cryptosporidium oocysts isolated during water monitoring.

    Science.gov (United States)

    Nichols, Rosely A B; Campbell, Brian M; Smith, Huw V

    2006-08-01

    We developed and validated a PCR-based method for identifying Cryptosporidium species and/or genotypes present on oocyst-positive microscope slides. The method involves removing coverslips and oocysts from previously examined slides followed by DNA extraction. We tested four loci, the 18S rRNA gene (N18SDIAG and N18SXIAO), the Cryptosporidium oocyst wall protein (COWP) gene (STN-COWP), and the dihydrofolate reductase (dhfr) gene (by multiplex allele-specific PCR), for amplifying DNA from low densities of Cryptosporidium parvum oocysts experimentally seeded onto microscope slides. The N18SDIAG locus performed consistently better than the other three tested. Purified oocysts from humans infected with C. felis, C. hominis, and C. parvum and commercially purchased C. muris were used to determine the sensitivities of three loci (N18SDIAG, STN-COWP, and N18SXIAO) to detect low oocyst densities. The N18SDIAG primers provided the greatest number of positive results, followed by the N18SXIAO primers and then the STN-COWP primers. Some oocyst-positive slides failed to generate a PCR product at any of the loci tested, but the limit of sensitivity is not entirely based on oocyst number. Sixteen of 33 environmental water monitoring Cryptosporidium slides tested (oocyst numbers ranging from 1 to 130) contained mixed Cryptosporidium species. The species/genotypes most commonly found were C. muris or C. andersoni, C. hominis or C. parvum, and C. meleagridis or Cryptosporidium sp. cervine, ferret, and mouse genotypes. Oocysts on one slide contained Cryptosporidium muskrat genotype II DNA.

  6. Transport of Cryptosporidium parvum Oocysts in a Silicon Micromodel

    Energy Technology Data Exchange (ETDEWEB)

    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.

  7. Characterization of an immunogenic glycocalyx on the surfaces of Cryptosporidium parvum oocysts and sporozoites.

    Science.gov (United States)

    Nanduri, J; Williams, S; Aji, T; Flanigan, T P

    1999-04-01

    Ruthenium red staining of Cryptosporidium parvum oocysts revealed the presence of a carbohydrate matrix on their outer bilayers that is characteristic of a glycocalyx. Surface labeling of intact oocysts identified material of high molecular weight (>10(6)) that reacted positively with sera from cryptosporidium-infected patients and with immunoglobulin A monoclonal antibodies.

  8. Characterization of an Immunogenic Glycocalyx on the Surfaces of Cryptosporidium parvum Oocysts and Sporozoites

    OpenAIRE

    Nanduri, Jayasri; Williams, Selvi; Aji, Toshiki; Flanigan, Timothy P.

    1999-01-01

    Ruthenium red staining of Cryptosporidium parvum oocysts revealed the presence of a carbohydrate matrix on their outer bilayers that is characteristic of a glycocalyx. Surface labeling of intact oocysts identified material of high molecular weight (>106) that reacted positively with sera from cryptosporidium-infected patients and with immunoglobulin A monoclonal antibodies.

  9. Viability Assessment of Cryptosporidium parvum Oocysts by Vital Dyes: Dry Mounts Overestimate the Number of “Ghost” Oocysts

    DEFF Research Database (Denmark)

    Petersen, Heidi Huus; Enemark, Heidi L.

    2017-01-01

    Viability assessment of Cryptosporidium parvum oocysts is crucial for evaluation of the public health significance of this important zoonotic protozoon. Viability is commonly assessed in wet mounts after acid pretreatmentand staining with fluorogenic vital dyes. However, in some studies, oocyst v...

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

    Directory of Open Access Journals (Sweden)

    Solo-Gabriele Helena María

    1998-01-01

    Full Text Available 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.

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

    Science.gov (United States)

    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. Published by Elsevier Ltd.

  12. Method to enumerate oocysts of cryptosporidium and cysts of Giardia in water

    International Nuclear Information System (INIS)

    Briancesco, R.; Bonadonna, L.

    2000-01-01

    Cryptosporidium and Giardia have been recognized as etiological agents of gastrointestinal illness in humans with severe consequences on children and immunocompromised individuals. Water seems to be vehicle of infection. In last years many efforts have been done to evaluate a method to enumerate oocysts of Cryptosporidium and cysts of Giardia in waters. Throughout filtration and concentration steps, the two procedures proposed allow to enumerate oocysts and cysts belonging to the two genera of protozoa [it

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

    DEFF Research Database (Denmark)

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

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

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

    Science.gov (United States)

    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.

  15. Detection of Cryptosporidium oocysts in green mussels (Perna viridis from shell-fish markets of Thailand

    Directory of Open Access Journals (Sweden)

    Srisuphanunt M.

    2009-09-01

    Full Text Available Mussels filter large volumes of water and can concentrate pathogenic organisms, which may act as potential vehicles of transmission to the consumer. A survey study was carried out to investigate the presence of Cryptosporidium protozoan parasites in green mussels (Perna viridis, the smussles pecies most destined for consumption in Thailand. In total, 56 samples were examined from Bangkok (n = 24 and Samut Prakan (n = 32 a wholesale shell-fish markets located at the mouth of the Chao Phraya River. The market for green mussels was closed to the mussel culture placed along the coastal line and this localization may have significant economical impact if the mussels’ cultures are found contaminated. Cryptosporidium spp. oocysts were detected by the immunofluorescence antibody method (IFA in 12.5% of the samples examined. The detection of Cryptosporidium oocysts in green mussels’ population of Samut Prakan was higher (15.6% than in Bangkok market (8.3%. These differences in positive samples from the two locations may be caused by physical, ecological and anthropogenic conditions. This could relay to different contamination levels of marine water by Cryptosporidium oocysts and consequently to contamination of harvested shellfish populations. The results demonstrate that the Cryptosporidium spp. oocysts were found indigenous in mussels from the coastal line of Thailand, indicating that mussels may act as a reservoir of Cryptosporidium foodborne infections for humans.

  16. Detection of Cryptosporidium oocysts in green mussels (Perna viridis) from shell-fish markets of Thailand.

    Science.gov (United States)

    Srisuphanunt, M; Wiwanitkit, Viroj; Saksirisampant, W; Karanis, P

    2009-09-01

    Mussels filter large volumes of water and can concentrate pathogenic organisms, which may act as potential vehicles of transmission to the consumer. A survey study was carried out to investigate the presence of Cryptosporidium protozoan parasites in green mussels (Perna viridis), the smussles pecies most destined for consumption in Thailand. In total, 56 samples were examined from Bangkok (n = 24) and Samut Prakan (n = 32) a wholesale shell-fish markets located at the mouth of the Chao Phraya River. The market for green mussels was closed to the mussel culture placed along the coastal line and this localization may have significant economical impact if the mussels' cultures are found contaminated. Cryptosporidium spp. oocysts were detected by the immunofluorescence antibody method (IFA) in 12.5% of the samples examined. The detection of Cryptosporidium oocysts in green mussels' population of Samut Prakan was higher (15.6%) than in Bangkok market (8.3%). These differences in positive samples from the two locations may be caused by physical, ecological and anthropogenic conditions. This could relay to different contamination levels of marine water by Cryptosporidium oocysts and consequently to contamination of harvested shellfish populations. The results demonstrate that the Cryptosporidium spp. oocysts were found indigenous in mussels from the coastal line of Thailand, indicating that mussels may act as a reservoir of Cryptosporidium foodborne infections for humans.

  17. New filtration system for efficient recovery of waterborne Cryptosporidium oocysts and Giardia cysts

    DEFF Research Database (Denmark)

    Al-Sabi, Mohammad Nafi Solaiman; Gad, J. A.; Riber, Ulla

    2015-01-01

    -)cysts (1x10(2); 10 replicates) was successfully amplified using real-time PCR.ConclusionsThe use of a metallic filter, sonication and air backwash' were key factors for creating a highly efficient system for recovery of apparently undamaged protozoa.Significance and Impact of the StudyThis reagent......AimsTo develop a filtration unit for efficient recovery of waterborne Cryptosporidium oocysts and Giardia cysts ((oo-)cysts) in drinking water.Methods and ResultsThis unit utilizes a metallic filter and an ultrasound transducer for eluting (oo-)cysts, with a fixed retentate backwash volume; approx....... 400l. Changes in the viability was evaluated by seeding wild type (oo-)cysts (1x10(4)) followed by sonication for 5, 10, 20 or 40s (five replicates for each period). Flow cytometry analysis showed negligible increase in the mortality of (oo-)cysts exposed to 5-10s of sonication. Recovery rate...

  18. Transport of Cryptosporidium parvum oocysts in soil columns following applications of raw and separated liquid slurries.

    Science.gov (United States)

    Petersen, Heidi H; Enemark, Heidi L; Olsen, Annette; Amin, M G Mostofa; Dalsgaard, Anders

    2012-09-01

    The potential for the transport of viable Cryptosporidium parvum oocysts through soil to land drains and groundwater was studied using simulated rainfall and intact soil columns which were applied raw slurry or separated liquid slurry. Following irrigation and weekly samplings over a 4-week period, C. parvum oocysts were detected from all soil columns regardless of slurry type and application method, although recovery rates were low (vertical distribution of oocysts, with more oocysts recovered from soil columns added liquid slurry irrespective of the irrigation status. Further studies are needed to determine the effectiveness of different slurry separation technologies to remove oocysts and other pathogens, as well as whether the application of separated liquid slurry to agricultural land may represent higher risks for groundwater contamination compared to application of raw slurry.

  19. Effectiveness of standard UV depuration at inactivating Cryptosporidium parvum recovered from spiked Pacific oysters (Crassostrea gigas).

    Science.gov (United States)

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

    2007-08-01

    When filter-feeding shellfish are consumed raw, because of their ability to concentrate and store waterborne pathogens, they are being increasingly associated with human gastroenteritis and have become recognized as important pathogen vectors. In the shellfish industry, UV depuration procedures are mandatory to reduce pathogen levels prior to human consumption. However, these guidelines are based around more susceptible fecal coliforms and Salmonella spp. and do not consider Cryptosporidium spp., which have significant resistance to environmental stresses. Thus, there is an urgent need to evaluate the efficiency of standard UV depuration against the survival of Cryptosporidium recovered from shellfish. Our study found that in industrial-scale shellfish depuration treatment tanks, standard UV treatment resulted in a 13-fold inactivation of recovered, viable C. parvum oocysts from spiked (1 x 10(6) oocysts liter (-1)) Pacific oysters. Depuration at half power also significantly reduced (P oysters. While UV treatment resulted in significant reductions of recovered viable oocysts, low numbers of viable oocysts were still recovered from oysters after depuration, making their consumption when raw a public health risk. Our study highlights the need for increased periodic monitoring programs for shellfish harvesting sites, improved depuration procedures, and revised microbial quality control parameters, including Cryptosporidium assessment, to minimize the risk of cryptosporidiosis.

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

    Science.gov (United States)

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

    2017-12-15

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

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

    Science.gov (United States)

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

  2. Gamma irradiation of Cryptosporidium parvum oocysts affects intracelluar levels of the viral symbiont CPV

    Science.gov (United States)

    Previous studies have shown a dose-dependent effect of gamma irradiation on Cryptosporidium parvum development in neonatal mice and newborn calves. In mice, C. parvum oocysts exposed to 200 Gy showed nearly complete inability to develop as measured by C. parvum-specific quantitative PCR of ileal ti...

  3. Effects of low and high temperatures on infectivity of Cryptosporidium muris oocysts suspended in water

    Czech Academy of Sciences Publication Activity Database

    Neumayerová, H.; Koudela, Břetislav

    2008-01-01

    Roč. 153, 3/4 (2008), s. 197-202 ISSN 0304-4017 Institutional research plan: CEZ:AV0Z60220518 Keywords : Cryptosporidium muris * oocysts * low and high temperatures * infectivity Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.039, year: 2008

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

    DEFF Research Database (Denmark)

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

    2016-01-01

    was carried out to investigate the effect of a coagulant produced from seeds of the Moringa oleifera tree (MO) in reducing Cryptosporidium parvum oocysts and turbidity in wastewater and stream water. Glass jars (n = 60) containing 500 mL wastewater obtained from the inlet to the primary settling tanks from...

  5. Occurrence of Cryptosporidium spp. oocysts in low quality water and on vegetables irrigated with low quality water in Kumasi, Ghana

    DEFF Research Database (Denmark)

    Petersen, Tobias B; Petersen, Heidi H.; Abaidoo, Robert C.

    Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp. contaminat......Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp...... but not on lettuce. Molecular characterization of Cryptosporidium positive samples was unsuccessful, thus no conclusions can be drawn concerning sources of contamination. Nevertheless, the detection of high prevalence and concentration levels of Cryptosporidium oocysts on vegetables consumed raw and in water...

  6. Fate of Cryptosporidium parvum oocysts within soil, water, and plant environment.

    Science.gov (United States)

    McLaughlin, Stephen J; Kalita, Prasanta K; Kuhlenschmidt, Mark S

    2013-12-15

    Vegetative Filter Strips (VFS) have long been used to control the movement of agricultural nutrients and prevent them from reaching receiving waters. Earlier studies have shown that VFS also dramatically reduce both the kinetics and extent of Cryptosporidium parvum (C. parvum) oocysts overland transport. In this study, we investigated possible mechanisms responsible for the ability of VFS to reduce oocyst overland transport. Measurement of the kinetics of C. parvum adhesion to individual sand, silt, and clay soil particles revealed that oocysts associate over time, albeit relatively slow, with clay but not silt or sand particles. Measurement of oocyst overland transport kinetics, soil infiltration depth, distance of travel, and adhesion to vegetation on bare and vegetated soil surfaces indicate that oocysts move more slowly, and penetrate the soil profile to a greater extent on a vegetated surface than on a bare soil surface. Furthermore, we demonstrate a small fraction of the oocysts become attached to vegetation at the soil-vegetation interface on VFS. These results suggest VFS function to reduce oocyst overland transport by primarily decreasing oocyst surface flow enough to allow penetration within the soil profile followed by subsequent adhesion to or entrapment within clay particle aggregates, and to a lesser extent, adhesion to the surface vegetation. Copyright © 2013 Elsevier Ltd. All rights reserved.

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

    DEFF Research Database (Denmark)

    Petersen, Heidi Huus; Enemark, Heidi L.; Olsen, Annette

    2012-01-01

    to determine the effectiveness of different slurry separation technologies to remove oocysts and other pathogens, as well as whether application of separated liquid slurry to agricultural land may represent higher risks for ground water contamination as compared to application of raw slurry.......The potential for transport of viable Cryptosporidium parvum oocysts through soil to land drains and groundwater was studied using simulated rainfall and intact soil columns which were applied raw slurry or separated liquid slurry. Following irrigation and weekly samplings over a four week period......, C. parvum oocysts were detected from all soil columns regardless of slurry type and application method although recovery rates were low (liquid slurry leached 73% and 90% more oocysts compared with columns with injected and surface applied raw slurry, respectively...

  8. Cryptosporidium oocysts in Ghanaian AIDS patients with diarrhoea ...

    African Journals Online (AJOL)

    Background: Although Cryptosporidium spp. infections in acquired immunodeficiency syndrome patients (AIDS) with chronic diarrhoea have been reported in several African countries, there is no information regarding cryptosporidial diarrhoea in Ghanaian AIDS patients. Objective: To investigate the occurrence of C.

  9. prevalence of cryptosporidium oocysts among children with acute

    African Journals Online (AJOL)

    USER

    2014-12-02

    Dec 2, 2014 ... disease caused by Cryptosporidium, is self-limiting gastroenteritis in the general ... is sufficient to produce infection and disease in susceptible hosts (Pereira et ..... 2006: Analysis of National Notification Data. Eurosurveillance.

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

    Directory of Open Access Journals (Sweden)

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

    2000-01-01

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

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

    Science.gov (United States)

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

    2005-01-01

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

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

    Science.gov (United States)

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

    2009-01-01

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

  13. Microscopic and Molecular Tracing of Cryptosporidium Oocysts: Identifying a Possible Reservoir of Infection in Red Grouse

    Directory of Open Access Journals (Sweden)

    David Baines

    2017-11-01

    Full Text Available Infection by Cryptosporidium baileyi causes respiratory cryptosporidiosis in red grouse Lagopus lagopus scotica. First diagnosed in 2010, it has since been detected across half of moors managed for grouse shooting in northern England. We hypothesised that contaminated grouse faeces within communal trays visited by grouse containing grit coated with flubendazole, provided to control Trichostrongylus tenuis parasites of grouse, is a reservoir of infection. To establish the basis to this hypothesis, contents of 23 trays from a grouse moor were examined for Cryptosporidium oocysts. Contents were subjected to Immuno Magnetic Separation oocyst concentration techniques prior to examination by Immuno Fluorescence Antibody Test microscopy and molecular analysis on the 18S rRNA gene. Seven of 13 (54% grit trays known to be used by infected grouse were positive for Cryptosporidium by IMS-IFAT, compared to two of 10 (20% random background trays. Ten of the 13 (77% trays used by infected birds amplified positive for Cryptosporidium by Polymerase Chain Reaction and three of the 10 (30% random trays. All PCR amplified products sequenced matched with C. baileyi, with C. parvum also present in one tray. These data suggest that trays used to “worm” grouse may act as reservoirs of Cryptosporidium infection and their future design may need to be reconsidered to minimise contamination.

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

    DEFF Research Database (Denmark)

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

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

  15. Evidence for a structural role for acid-fast lipids in oocyst walls of Cryptosporidium, Toxoplasma, and Eimeria.

    Science.gov (United States)

    Bushkin, G Guy; Motari, Edwin; Carpentieri, Andrea; Dubey, Jitender P; Costello, Catherine E; Robbins, Phillips W; Samuelson, John

    2013-09-03

    Coccidia are protozoan parasites that cause significant human disease and are of major agricultural importance. Cryptosporidium spp. cause diarrhea in humans and animals, while Toxoplasma causes disseminated infections in fetuses and untreated AIDS patients. Eimeria is a major pathogen of commercial chickens. Oocysts, which are the infectious form of Cryptosporidium and Eimeria and one of two infectious forms of Toxoplasma (the other is tissue cysts in undercooked meat), have a multilayered wall. Recently we showed that the inner layer of the oocyst walls of Toxoplasma and Eimeria is a porous scaffold of fibers of β-1,3-glucan, which are also present in fungal walls but are absent from Cryptosporidium oocyst walls. Here we present evidence for a structural role for lipids in the oocyst walls of Cryptosporidium, Toxoplasma, and Eimeria. Briefly, oocyst walls of each organism label with acid-fast stains that bind to lipids in the walls of mycobacteria. Polyketide synthases similar to those that make mycobacterial wall lipids are abundant in oocysts of Toxoplasma and Eimeria and are predicted in Cryptosporidium. The outer layer of oocyst wall of Eimeria and the entire oocyst wall of Cryptosporidium are dissolved by organic solvents. Oocyst wall lipids are complex mixtures of triglycerides, some of which contain polyhydroxy fatty acyl chains like those present in plant cutin or elongated fatty acyl chains like mycolic acids. We propose a two-layered model of the oocyst wall (glucan and acid-fast lipids) that resembles the two-layered walls of mycobacteria (peptidoglycan and acid-fast lipids) and plants (cellulose and cutin). Oocysts, which are essential for the fecal-oral spread of coccidia, have a wall that is thought responsible for their survival in the environment and for their transit through the stomach and small intestine. While oocyst walls of Toxoplasma and Eimeria are strengthened by a porous scaffold of fibrils of β-1,3-glucan and by proteins cross

  16. Contamination microbiologique des eaux souterraines par les oocystes de Cryptosporidium en Haïti. Evaluation des risques pour la santé de la population

    OpenAIRE

    Balthazard-Accou , Ketty; Emmanuel , Evens; Diouf , Momar; Agnamey , Patrice

    2017-01-01

    International audience; Contamination of natural aquatic ecosystems by Cryptosporidium is a major environmental and human health issue. In Haiti, environmental Cryptosporidium oocysts pollution has been well documented by previous studies conducted in several cities of the country. In groundwater from Les Cayes of Haiti, significant concentrations from 1 to 989 oocysts in 100 liters of filtered water were calculated. Results of these studies revealed high level of Cryptosporidium oocysts poll...

  17. Statistical comparison of excystation methods in Cryptosporidium parvum oocysts

    Czech Academy of Sciences Publication Activity Database

    Pecková, R.; Stuart, P. D.; Sak, Bohumil; Květoňová, Dana; Kváč, Martin; Foitová, I.

    2016-01-01

    Roč. 230, OCT 30 (2016), s. 1-5 ISSN 0304-4017 R&D Projects: GA ČR(CZ) GAP505/11/1163 Institutional support: RVO:60077344 Keywords : Cryptosporidium parvum * excystation methods * in vitro cultivation * sodium hypochlorite * tlypsin Subject RIV: EG - Zoology Impact factor: 2.356, year: 2016

  18. prevalence of cryptosporidium oocysts among children with acute

    African Journals Online (AJOL)

    USER

    2014-12-02

    Dec 2, 2014 ... Cryptosporidium, a coccidian protozoan parasite, is an important causative agent of human and animal gastrointestinal illness globally (Huang et al., 2009). ..... person, animal-to-person, waterborne, food-borne, and possible airborne transmission. This need to be further investigated. The observation made ...

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

    Directory of Open Access Journals (Sweden)

    Asma Iqbal

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

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

    DEFF Research Database (Denmark)

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

    The widespread waterborne pathogen Cryptosporidium parvum is primarily transmitted to humans via contaminated drinking and recreational water. Nearly all drinking water in Denmark is groundwater, but this can be contaminated with oocysts from application of contaminated manure to the field. Oocysts...... in the leachates from soil columns to which Cryptosporidium positive slurry had been injected. Although recovery rates were low, regardless of slurry type, C. parvum oocysts were detected from all soil columns. Variations in the leachate patterns were recorded between soil columns added raw and liquid slurry...

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

    Directory of Open Access Journals (Sweden)

    Christophe J. G. Darnault

    2017-06-01

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

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

    Science.gov (United States)

    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

  3. Validation of a new technique to detect Cryptosporidium spp. oocysts in bovine feces.

    Science.gov (United States)

    Inácio, Sandra Valéria; Gomes, Jancarlo Ferreira; Oliveira, Bruno César Miranda; Falcão, Alexandre Xavier; Suzuki, Celso Tetsuo Nagase; Dos Santos, Bianca Martins; de Aquino, Monally Conceição Costa; de Paula Ribeiro, Rafaela Silva; de Assunção, Danilla Mendes; Casemiro, Pamella Almeida Freire; Meireles, Marcelo Vasconcelos; Bresciani, Katia Denise Saraiva

    2016-11-01

    Due to its important zoonotic potential, cryptosporidiosis arouses strong interest in the scientific community, because, it was initially considered a rare and opportunistic disease. The parasitological diagnosis of the causative agent of this disease, the protozoan Cryptosporidium spp., requires the use of specific techniques of concentration and permanent staining, which are laborious and costly, and are difficult to use in routine laboratory tests. In view of the above, we conducted the feasibility, development, evaluation and intralaboratory validation of a new parasitological technique for analysis in optical microscopy of Cryptosporidium spp. oocysts, called TF-Test Coccidia, using fecal samples from calves from the city of Araçatuba, São Paulo. To confirm the aforementioned parasite and prove the diagnostic efficiency of the new technique, we used two established methodologies in the scientific literature: parasite concentration by centrifugal sedimentation and negative staining with malachite green (CSN-Malachite) and Nested-PCR. We observed good effectiveness of the TF-Test Coccidia technique, being statistically equivalent to CSN-Malachite. Thus, we verified the effectiveness of the TF-Test Coccidia parasitological technique for the detection of Cryptosporidium spp. oocysts and observed good concentration and morphology of the parasite, with a low amount of debris in the fecal smear. Copyright © 2016 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2007-01-01

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

  5. Studies on the resistance/reactivation of Giardia muris cysts and Cryptosporidium parvum oocysts exposed to medium-pressure ultraviolet radiation.

    Science.gov (United States)

    Belosevic, M; Craik, S A; Stafford, J L; Neumann, N F; Kruithof, J; Smith, D W

    2001-10-16

    The ex vivo and in vivo reactivation of Giardia muris cysts and Cryptosporidium parvum oocysts after exposure to different doses of ultraviolet (UV) radiation was determined using animal infectivity. The infectivity of UV-treated parasites stored for 1-4 days (G. muris) or 1-17 days (C. parvum) at room temperature in the dark was similar to that of organisms administered immediately after UV treatment, indicating that the parasites did not reactivate ex vivo. In contrast, we observed in vivo reactivation of G. muris in three of seven independent animal infectivity experiments, when parasites were treated with relatively low doses of medium-pressure UV (muris cysts and C. parvum oocysts exposed to medium-pressure UV doses of 60 mJ/cm(2) or higher did not exhibit resistance to and/or reactivation following treatment. This suggests that when appropriate doses of UV are used, significant and permanent inactivation of these parasites may be achieved.

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

    Science.gov (United States)

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

    2017-12-01

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

  7. New system for higher recovery rate of water borne Cryptosporidium oocysts and Giardia cysts

    DEFF Research Database (Denmark)

    Al-Sabi, Mohammad Nafi Solaiman; Gad, Jens; Klinting, Mette

    2012-01-01

    Background: The two most common water borne pathogenic protozoa, Cryptosporidium and Giardia, cause diarrhea worldwide. Detecting these parasites in water samples depends on effective parasite recovery from the water matrix. The reported low recovery rates of the currently used filter methods...... motivate the development of systems with higher recovery rates. Materials and methods: Five replicates of IMS purified Cryptosporidium oocysts and Giardia cysts (N=2x103) were injected into a specially coated filter unit with a carefully chosen pore size. Following filtration, sonication was performed...... were 85% were recorded when the filter was sonicated. Sonication usually affects parasite viability but could be tuned into a useful tool for enhanced backwash collection of parasites using a specially constructed filter unit and a sonication protocol. The filtration...

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

    DEFF Research Database (Denmark)

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

    The widespread waterborne pathogen Cryptosporidium parvum is frequently transmitted to humans via contaminated drinking and recreational water. Nearly all drinking water in Denmark is groundwater, which can be contaminated with oocysts e.g. from application of contaminated manure to the field...... in the leachates from soil columns to which Cryptosporidium positive slurry had been injected. Although recovery rates were low, regardless of slurry type, C. parvum oocysts were detected from all soil columns. Variations in the leachate patterns were recorded between soil columns added raw and liquid slurry...

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

    Science.gov (United States)

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

    2015-05-15

    Cryptosporidium continues to be problematic for the water industry, with risk assessments often indicating that treatment barriers may fail under extreme conditions. However, risk analyses have historically used oocyst densities and not considered either oocyst infectivity or species/genotype, which can result in an overestimation of risk if the oocysts are not human infective. We describe an integrated assay for determining oocyst density, infectivity, and genotype from a single-sample concentrate, an important advance that overcomes the need for processing multiple-grab samples or splitting sample concentrates for separate analyses. The assay incorporates an oocyst recovery control and is compatible with standard primary concentration techniques. Oocysts were purified from primary concentrates using immunomagnetic separation prior to processing by an infectivity assay. Plate-based cell culture was used to detect infectious foci, with a monolayer washing protocol developed to allow recovery and enumeration of oocysts. A simple DNA extraction protocol was developed to allow typing of any wells containing infectious Cryptosporidium. Water samples from a variety of source water and wastewater matrices, including a semirural catchment, wastewater, an aquifer recharge site, and storm water, were analyzed using the assay. Results demonstrate that the assay can reliably determine oocyst densities, infectivity, and genotype from single-grab samples for a variety of water matrices and emphasize the varying nature of Cryptosporidium risk extant throughout source waters and wastewaters. This assay should therefore enable a more comprehensive understanding of Cryptosporidium risk for different water sources, assisting in the selection of appropriate risk mitigation measures. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  10. An easy 'one tube' method to estimate viability of Cryptosporidium oocysts using real-time qPCR

    NARCIS (Netherlands)

    Paziewska-Harris, A.; Schoone, G.; Schallig, H. D. F. H.

    2016-01-01

    Viability estimation of the highly resistant oocysts of Cryptosporidium remains a key issue for the monitoring and control of this pathogen. We present here a simple 'one tube' quantitative PCR (qPCR) protocol for viability estimation using a DNA extraction protocol which preferentially solubilizes

  11. Techniques for the recovery and identification of Cryptosporidium oocysts from stool specimens.

    Science.gov (United States)

    Garcia, L S; Bruckner, D A; Brewer, T C; Shimizu, R Y

    1983-07-01

    Due to increasing numbers of patients with documented infections with Cryptosporidium and other coccidia, it is important for the physician and clinical laboratory to be aware of the appropriate diagnostic techniques necessary for organism recovery and identification. Although Cryptosporidium is found in the gastrointestinal tract, tissue biopsies may be insufficient for organism recovery; the examination of stool specimens is a noninvasive procedure and will provide better overall opportunities for organism recovery. Human clinical specimens were examined from 45 patients with confirmed cryptosporidiosis or suspected of having the infection. Tissue biopsy sections, fecal wet preparations, and permanent stained smears were examined. Stool specimens were submitted in 10% Formalin, 2.5% potassium dichromate, and polyvinyl alcohol and were examined for oocysts by using 15 different methods: phase-contrast and light microscopy; Sheather's sugar flotation; Formalin concentration techniques; 10% potassium hydroxide; Giemsa; trichrome; periodic acid-Schiff; modified periodic acid-Schiff; silver methenamine; acridine orange; auramine-rhodamine; Kinyoun acid-fast; Ziehl-Neelsen carbolfuchsin; and a modified acid-fast procedure. Each technique or combination of techniques was assessed by organism quantitation, organism morphology, and ease of visual recognition. Based on these comparative studies, the modified Ziehl-Neelsen carbolfuchsin stain on 10% Formalin-preserved stool is recommended for the recovery and identification of Cryptosporidium.

  12. Detection of Cryptosporidium and Cyclospora Oocysts from Environmental Water for Drinking and Recreational Activities in Sarawak, Malaysia.

    Science.gov (United States)

    Bilung, Lesley Maurice; Tahar, Ahmad Syatir; Yunos, Nur Emyliana; Apun, Kasing; Lim, Yvonne Ai-Lian; Nillian, Elexson; Hashim, Hashimatul Fatma

    2017-01-01

    Cryptosporidiosis and cyclosporiasis are caused by waterborne coccidian protozoan parasites of the genera Cryptosporidium and Cyclospora, respectively. This study was conducted to detect Cryptosporidium and Cyclospora oocysts from environmental water abstracted by drinking water treatment plants and recreational activities in Sarawak, Malaysia. Water samples (12 each) were collected from Sungai Sarawak Kanan in Bau and Sungai Sarawak Kiri in Batu Kitang, respectively. In addition, 6 water samples each were collected from Ranchan Recreational Park and UNIMAS Lake at Universiti Malaysia Sarawak, Kota Samarahan, respectively. Water physicochemical parameters were also recorded. All samples were concentrated by the iron sulfate flocculation method followed by the sucrose floatation technique. Cryptosporidium and Cyclospora were detected by modified Ziehl-Neelsen technique. Correlation of the parasites distribution with water physicochemical parameters was analysed using bivariate Pearson correlation. Based on the 24 total samples of environmental water abstracted by drinking water treatment plants, all the samples (24/24; 100%) were positive with Cryptosporidium , and only 2 samples (2/24; 8.33%) were positive with Cyclospora . Based on the 12 total samples of water for recreational activities, 4 samples (4/12; 33%) were positive with Cryptosporidium , while 2 samples (2/12; 17%) were positive with Cyclospora . Cryptosporidium oocysts were negatively correlated with dissolved oxygen (DO).

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

    Science.gov (United States)

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

    2008-12-01

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

  14. Occurrence of Giardia cysts and Cryptosporidium oocysts in activated sludge samples in Campinas, SP, Brazil.

    Science.gov (United States)

    Santos, Luciana Urbano; Bonatti, Taís Rondello; Cantusio Neto, Romeu; Franco, Regina Maura Bueno

    2004-01-01

    Giardia and Cryptosporidium have caused several outbreaks of gastroenteritis in humans associated with drinking water. Contaminated sewage effluents are recognized as a potential source of waterborne protozoa. Due to the lack of studies about the occurrence of these parasites in sewage samples in Brazil, we compared the efficiency of two procedures for concentrating cysts and oocysts in activated sludge samples of one sewage treatment plant. For this, the samples were submitted to i) concentration by the ether clarification procedure (ECP) and to ii) purification by sucrose flotation method (SFM) and aliquots of the pellets were examined by immunofluorescence. Giardia cysts were present in all samples (100.0%; n = 8) when using ECP and kit 1 reagents, while kit 2 resulted in six positive samples (85.7%; n = 7). As for SFM, cysts were detected in 75.0% and 100.0% of these samples (for kit 1 and 2, respectively). Regarding Cryptosporidium, two samples (25.0%; kit 1 and 28.5% for kit 2) were detected positive by using ECP, while for SFM, only one sample (examined by kit 1) was positive (12.5%). The results of the control trial revealed Giardia and Cryptosporidium recovery efficiency rates for ECP of 54.5% and 9.6%, while SFM was 10.5% and 3.2%, respectively. Considering the high concentration detected, a previous evaluation of the activated sludge before its application in agriculture is recommended and with some improvement, ECP would be an appropriate simple technique for protozoa detection in sewage samples.

  15. Detection of infectious Cryptosporidium oocysts by cell culture: applicability to environmental samples

    NARCIS (Netherlands)

    Schets FM; Engels GB; During M; Roda Husman AM de; MGB

    2004-01-01

    Cryptosporidium is one of the important causative agents of gastrointestinal illness in humans. Cryptosporidium infections are often waterborne and can be transmitted through drinking water or recreational water. Estimation of the risk of infection with Cryptosporidium after exposure to drinking

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

    Science.gov (United States)

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

    2010-10-01

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

  17. Detection of infectious Cryptosporidium oocysts by cell culture: applicability to environmental samples

    NARCIS (Netherlands)

    Schets FM; Engels GB; During M; de Roda Husman AM; MGB

    2004-01-01

    Cryptosporidium is een van de belangrijkste veroorzakers van gastro-enteritis bij de mens. Cryptosporidium-infecties worden vaak via water overgedragen, dit kan zowel drinkwater als recreatiewater zijn. Bij schatting van de kans op infectie met Cryptosporidium na blootstelling aan drinkwater is

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

    Science.gov (United States)

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

    2012-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Sadia Benamrouz

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

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

    Science.gov (United States)

    Benamrouz, Sadia; Guyot, Karine; Gazzola, Sophie; Mouray, Anthony; Chassat, Thierry; Delaire, Baptiste; Chabé, Magali; Gosset, Pierre; Viscogliosi, Eric; Dei-Cas, Eduardo; Creusy, Colette; Conseil, Valerie; Certad, Gabriela

    2012-01-01

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

  1. Inactivation credit of UV radiation for viruses, bacteria and protozoan (oo)cysts in water: a review

    NARCIS (Netherlands)

    Hijnen, W.A.M.; Beerendonk, E.F.; Medema, Gerriet Jan

    2006-01-01

    UV disinfection technology is of growing interest in the water industry since it was demonstrated that UV radiation is very effective against (oo)cysts of Cryptosporidium and Giardia, two pathogenic micro-organisms of major importance for the safety of drinking water. Quantitative Microbial Risk

  2. Fluorescent microspheres as surrogates in evaluating the efficacy of riverbank filtration for removing Cryptosporidium parvum oocysts and other pathogens

    Science.gov (United States)

    Harvey, Ronald W.; Metge, David W.; Sheets, Rodney A.; Jasperse, Jay

    2011-01-01

    A major benefit of riverbank filtration (RBF) is that it provides a relatively effective means for pathogen removal. There is a need to conduct more injection-and-recovery transport studies at operating RBF sites in order to properly assess the combined effects of the site heterogeneities and ambient physicochemical conditions, which are difficult to replicate in the lab. For field transport studies involving pathogens, there is considerable interest in using fluorescent carboxylated microspheres (FCM) as surrogates, because they are chemically inert, negatively charged, easy to detect, available in a wide variety of sizes, and have been found to be nonhazardous in tracer applications. Although there have been a number of in-situ studies comparing the subsurface transport behaviors of FCM to those of bacteria and viruses, much less is known about their suitability for investigations of protozoa. Oocysts of the intestinal protozoan pathogen Cryptosporidium spp are of particular concern for many RBF operations because of their ubiquity and persistence in rivers and high resistance to chlorine disinfection. Although microspheres often have proven to be less-than-ideal analogs for capturing the abiotic transport behavior of viruses and bacteria, there is encouraging recent evidence regarding use of FCM as surrogates for C. parvum oocysts. This chapter discusses the potential of fluorescent microspheres as safe and easy-to-detect surrogates for evaluating the efficacy of RBF operations for removing pathogens, particularly Cryptosporidium, from source waters at different points along the flow path.

  3. Identification and determination of the viability of Giardia lamblia cysts and Cryptosporidium parvum and Cryptosporidium hominis oocysts in human fecal and water supply samples by fluorescent in situ hybridization (FISH) and monoclonal antibodies.

    Science.gov (United States)

    Lemos, Vanessa; Graczyk, Thaddeus K; Alves, Margarida; Lobo, Maria Luísa; Sousa, Maria C; Antunes, Francisco; Matos, Olga

    2005-12-01

    In the present study, fluorescent in situ hybridization (FISH) and monoclonal antibodies (MAbs) were evaluated for species-specific detection and viability determination of Giardia lamblia, Cryptosporidium parvum, and Cryptosporidium hominis in human fecal and water supply samples. A total of 50 fecal human samples positive for G. lamblia cysts, 38 positive for C. parvum, and 23 positive for C. hominis were studied. Also, 18 water supply samples positive for Giardia spp. and Cryptosporidium spp. by the United States Environmental Protection Agency (USEPA) Method 1623 were studied by FISH and fluorescein isothiocyanate (FITC)-conjugated MAbs. Eighteen percent of the fecal samples parasitologically positive for G. lamblia presented viable and nonviable cysts, and 5% of those positive for Cryptosporidium spp. presented viable and nonviable oocysts. Of the 18 water supply samples analyzed, 6 (33%) presented Giardia spp. viable and nonviable cysts and 2 (11%) presented viable and nonviable Cryptosporidium spp. oocysts. G. lamblia identification was confirmed by polymerase chain reaction (PCR) and sequencing of the beta-giardin gene in the fecal and water samples found positive by FISH and FITC-conjugated MAbs. C. parvum and Cryptosporidium muris were identified, by PCR and sequencing of the small subunit of ribosomal RNA gene, in seven and one water samples, respectively. Our results confirm that this technique enables simultaneous visualization, species-specific identification, and viability determination of the organisms present in human fecal and water supply samples.

  4. Assessing viability and infectivity of foodborne and waterborne stages (cysts/oocysts of Giardia duodenalis, Cryptosporidium spp., and Toxoplasma gondii: a review of methods

    Directory of Open Access Journals (Sweden)

    Rousseau Angélique

    2018-01-01

    Full Text Available Giardia duodenalis, Cryptosporidium spp. and Toxoplasma gondii are protozoan parasites that have been highlighted as emerging foodborne pathogens by the Food and Agriculture Organization of the United Nations and the World Health Organization. According to the European Food Safety Authority, 4786 foodborne and waterborne outbreaks were reported in Europe in 2016, of which 0.4% were attributed to parasites including Cryptosporidium, Giardia and Trichinella. Until 2016, no standardized methods were available to detect Giardia, Cryptosporidium and Toxoplasma (oocysts in food. Therefore, no regulation exists regarding these biohazards. Nevertheless, considering their low infective dose, ingestion of foodstuffs contaminated by low quantities of these three parasites can lead to human infection. To evaluate the risk of protozoan parasites in food, efforts must be made towards exposure assessment to estimate the contamination along the food chain, from raw products to consumers. This requires determining: (i the occurrence of infective protozoan (oocysts in foods, and (ii the efficacy of control measures to eliminate this contamination. In order to conduct such assessments, methods for identification of viable (i.e. live and infective parasites are required. This review describes the methods currently available to evaluate infectivity and viability of G. duodenalis cysts, Cryptosporidium spp. and T. gondii oocysts, and their potential for application in exposure assessment to determine the presence of the infective protozoa and/or to characterize the efficacy of control measures. Advantages and limits of each method are highlighted and an analytical strategy is proposed to assess exposure to these protozoa.

  5. BLIND TRIALS EVALUATING IN VITRO INFECTIVITY OF CRYPTOSPORIDIUM PARVUM OOCYSTS USING CELL CULTURE IMMUNOFLUORESCENCE

    Science.gov (United States)

    An optimized cell culture-immunofluorescence (IFA) procedure, using the HCT-8 cell line, was evaluated in 'blind' trials to determine the sensitivity and reproducibility for measuring infectivity of flow cytometry prepared inocula of C. parvum oocysts. In separate trials, suspens...

  6. Studies on the prevalence of giardia cysts and cryptosporidium oocysts in South-African water

    CSIR Research Space (South Africa)

    Kfir, R

    1995-01-01

    Full Text Available per sample and the number of positive samples. Almost 50% of sewage samples studied contained Giardia cysts and 30% contained both Giardia cysts and Cryptosporium oocysts. Treatment of sewage resulted in a reduction in the percentage of samples...

  7. Viability staining and animal infectivity of Cryptosporidium andersoni oocysts after long-term storage

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Květoňová, Dana; Salát, Jiří; Ditrich, Oleg

    2007-01-01

    Roč. 100, č. 2 (2007), s. 213-217 ISSN 0932-0113 R&D Projects: GA ČR GA524/05/0992 Institutional research plan: CEZ:AV0Z60220518 Keywords : Cryptosporidium and ersoni * viability * infectivity * long-term storage Subject RIV: EG - Zoology Impact factor: 1.512, year: 2007

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2013-02-04

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

  9. The role of free-ranging, captive, and domestic birds of Western Poland in environmental contamination with Cryptosporidium parvum oocysts and Giardia lamblia cysts.

    Science.gov (United States)

    Majewska, Anna C; Graczyk, Thaddeus K; Słodkowicz-Kowalska, Anna; Tamang, Leena; Jedrzejewski, Szymon; Zduniak, Piotr; Solarczyk, Piotr; Nowosad, Andrzej; Nowosad, Piotr

    2009-04-01

    As Cryptosporidium parvum and Giardia lamblia can be disseminated in the environment by avian hosts, a total of 499 fecal dropping from 308 free-ranging, 90 captive, and 101 domestic birds were tested by conventional, immunological, and molecular techniques for these human enteropathogens. Twenty-six (5.2%) tested positive for G. lamblia cysts and 19 (3.8%) for C. parvum oocysts. A bird total of 23 (7.5%) free-ranging, two (2.2%) captive, and one (0.1%) domestic tested positive for cysts, whereas 18 (5.8%) free-ranging, one (1.1%) captive, and zero livestock birds tested positive for oocysts. G. lamblia cysts and C. parvum oocysts were found significantly more frequently in fecal droppings of free-ranging aquatic birds than in birds not normally associated with water. No specimen tested positive for both pathogens simultaneously. Aquatic birds represent an important epidemiologic link in water-associated transmission cycles of Cryptosporidium and Giardia and play a significant role in environmental contamination of aquatic habitats with these anthropozoonotic pathogens.

  10. Effects of Ultrasound on the Survival and Characteristics of Cryptosporidium Oocysts and Giardia Cysts

    DEFF Research Database (Denmark)

    Al-Sabi, Mohammad Nafi Solaiman; Gad, J.; Klinting, M.

    2011-01-01

    determine the effects of ultrasound on the parasite, including the sonication power of ultrasound as well as substrate temperature. Conclusions: Ultrasound is harmful for waterborne protozoa even when momentarily applied. However, a mode of operation may exist in which ultrasound can be used for collection......-. Additionally ultrasound has been used for cleaning filters used for water sampling and purification. Other studies have shown that backwash sampling of filtrates, and thereby collection of microorganisms, can be facilitated by sonication. Methods: We studied the effects of ultrasound with different sonication...... power and time durations on two of the most common waterborne protozoa Cryptosporidium and Giardia, and examined its effects on parasite characteristics and survival rate using immunofluorescence dyes; DAPI (4’,6-diamidino-2-phenylindol) staining/PI (propidium iodide), and analyzed by flow cytometry...

  11. First report of predation of Giardia sp. cysts by ciliated protozoa and confirmation of predation of Cryptosporidium spp. oocysts by ciliate species.

    Science.gov (United States)

    Siqueira-Castro, Isabel Cristina Vidal; Greinert-Goulart, Juliane Araújo; Bonatti, Tais Rondello; Yamashiro, Sandra; Franco, Regina Maura Bueno

    2016-06-01

    Ciliated protozoa are important components of the microbial food web in various habitats, especially aquatic environments. These organisms are useful bioindicators for both environmental quality assessment and the wastewater purification process. The pathogenic parasitic protozoan species Giardia and Cryptosporidium represent a significant concern for human health, being responsible for numerous disease outbreaks worldwide. The predation of cysts and oocysts in 15 ciliate species from water and sewage samples collected in Campinas, São Paulo, Brazil were verified under laboratory conditions. The ciliated protozoan species were selected based on their mode of nutrition, and only bacterivorous and suspension-feeders were considered for the experiments. The species Blepharisma sinuosum, Euplotes aediculatus, Sterkiella cavicola, Oxytricha granulifera, Vorticella infusionum, Spirostomum minus, and Stentor coeruleus ingested cysts and oocysts, the resistance forms of Giardia spp. and Cryptosporidium spp., respectively. This is the first time that the ingestion of Giardia cysts by ciliated protozoa has been reported. These findings may contribute to a better understanding of the biological removal of these pathogens from aquatic environments.

  12. The pulsed light inactivation of veterinary relevant microbial biofilms ...

    African Journals Online (AJOL)

    Results show that both Cryptosporidium and Giardia attach to biofilms in large numbers (100-1000 oo/cysts) in as little as 72 hours. Pulsed light successfully inactivated all test species (Listeria, Salmonella, Bacillus, Escherichia) in planktonic and biofilm form with an increase in inactivation for every increase in UV dose.

  13. Occurrence of Giardia cysts and Cryptosporidium oocysts in activated sludge samples in Campinas, SP, Brazil Ocorrência de cistos de Giardia e oocistos de Cryptosporidium em amostras de lodo ativado em Campinas, SP, Brasil

    Directory of Open Access Journals (Sweden)

    Luciana Urbano Santos

    2004-04-01

    Full Text Available Giardia and Cryptosporidium have caused several outbreaks of gastroenteritis in humans associated with drinking water. Contaminated sewage effluents are recognized as a potential source of waterborne protozoa. Due to the lack of studies about the occurrence of these parasites in sewage samples in Brazil, we compared the efficiency of two procedures for concentrating cysts and oocysts in activated sludge samples of one sewage treatment plant. For this, the samples were submitted to i concentration by the ether clarification procedure (ECP and to ii purification by sucrose flotation method (SFM and aliquots of the pellets were examined by immunofluorescence. Giardia cysts were present in all samples (100.0%; n = 8 when using ECP and kit 1 reagents, while kit 2 resulted in six positive samples (85.7%; n = 7. As for SFM, cysts were detected in 75.0% and 100.0% of these samples (for kit 1 and 2, respectively. Regarding Cryptosporidium, two samples (25.0%; kit 1 and 28.5% for kit 2 were detected positive by using ECP, while for SFM, only one sample (examined by kit 1 was positive (12.5%. The results of the control trial revealed Giardia and Cryptosporidium recovery efficiency rates for ECP of 54.5% and 9.6%, while SFM was 10.5% and 3.2%, respectively. Considering the high concentration detected, a previous evaluation of the activated sludge before its application in agriculture is recommended and with some improvement, ECP would be an appropriate simple technique for protozoa detection in sewage samples.Giardia e Cryptosporidium causaram vários surtos epidêmicos de gastroenterite, associados à água potável. Efluentes de esgoto contaminados foram incriminados como uma fonte potencial de cistos e oocistos. Uma investigação foi conduzida para verificar a presença de cistos de Giardia e oocistos de Cryptosporidium em amostras de lodo ativado de uma Estação de Tratamento de Esgoto. Para isto as amostras foram submetidas: i a concentração pelo

  14. Cryptosporidium Attenuation across the Wastewater Treatment Train: Recycled Water Fit for Purpose.

    Science.gov (United States)

    King, Brendon; Fanok, Stella; Phillips, Renae; Lau, Melody; van den Akker, Ben; Monis, Paul

    2017-03-01

    Compliance with guideline removal targets for Cryptosporidium which do not provide any credit for the inactivation of oocysts through wastewater treatment processes can considerably increase the cost of providing recycled water. Here we present the application of an integrated assay to quantify both oocyst numbers and infectivity levels after various treatment stages at three Victorian and two South Australian (SA) wastewater treatment plants (WWTPs). Oocyst density in the raw sewage was commensurate with community disease burden, with early rounds of sampling capturing a widespread cryptosporidiosis outbreak in Victoria. The level of infectivity of oocysts in sewage was stable throughout the year but was significantly lower at the SA WWTPs. Removals across secondary treatment processes were seasonal, with poorer removals associated with inflow variability; however, no decrease in the oocyst infectivity was identified. For SA WWTPs, those oocysts remaining within the secondary treatment-clarified effluent were proportionally more infectious than those in raw sewage. Lagoon systems demonstrated significant inactivation or removal of oocysts, with attenuation being seasonal. Examination of a UV system emphasized its efficacy as a disinfectant barrier but conversely confirmed the importance of a multibarrier approach with the detection of infectious oocysts postdisinfection. The ability to characterize risk from infectious oocysts revealed that the risk from Cryptosporidium is significantly lower than previously thought and that its inclusion in quantitative risk assessments of reuse systems will more accurately direct the selection of treatment strategies and capital expenditure, influencing the sustainability of such schemes. IMPORTANCE Here we present the application of a recently developed integrated assay not only to quantify the removal of Cryptosporidium oocysts but also to quantify their infectivity across various treatment stages at five wastewater treatment

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

    Science.gov (United States)

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

    2015-01-01

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

  16. Detecção de oocistos de Cryptosporidium spp. e cistos de Giardia spp. em amostras de esgoto bruto ou tratado: avaliação crítica dos métodos Detection of Cryptosporidium spp. oocysts and Giardia spp. cysts in raw and effluent wastewater: critical evaluation of methods

    Directory of Open Access Journals (Sweden)

    Luciana Urbano dos Santos

    2011-06-01

    Full Text Available Neste trabalho, avaliou-se a eficiência dos métodos centrífugo-concentração e filtração em membrana, na detecção de oocistos de Cryptosporidium spp. e cistos de Giardia spp. em amostras de esgoto bruto e tratado, provenientes de um sistema de lodos ativados (estação de tratamento de esgoto, Samambaia, Campinas, em São Paulo. As amostras foram coletadas quinzenalmente por dois anos: 53 amostras de esgoto bruto (AFL, 53 de efluente tratado sem desinfecção por luz ultravioleta (EFL e 38 de efluente tratado e desinfetado por luz ultravioleta (EFL+UV. Cistos de Giardia spp. foram encontrados em 90,5% das amostras AFL; em 96,2%, de EFL; e em 94,7%, de EFL+UV. Oocistos de Cryptosporidium spp. foram detectados em 6,4% das amostras AFL e em 2,6 % de EFL+UV. Ambos os métodos mostraram-se eficientes na detecção destes protozoários em todos os tipos de amostras, além de apresentarem baixo custo por análise.In this study, the efficiency of centrifuge-concentration and membrane filtrated methods was evaluated in the detection of Cryptosporidium spp. oocysts and Giardia spp. cysts in raw or treated wastewater samples, from activated sludge systems (ETE - Samambaia, Campinas, in São Paulo. The samples were collected once a fortnight for two years: 53 samples of influent (AFL, 53 samples of treated effluent without ultraviolet disinfection (EFL, and 38 samples of treated effluent with ultraviolet disinfection (EFL+UV. Giardia spp. cysts were found in 90.5% of the AFL samples; in 96.2% of the samples, EFL; and in 94.7%, EFL+UV. Cryptosporidium spp. oocysts were detected in 6.4% of AFL samples and 2.6% of EFL+UV. Both methods showed efficiency when detecting protozoa in all types of samples, besides having low costs by analysis.

  17. Disinfection technology with ozone for cryptosporidium; Cryptosporidium taisaku to shite no ozone shodoku gijutsu

    Energy Technology Data Exchange (ETDEWEB)

    Tanaka, Y.; Takahashi, K. [Fuji Electric Co. Ltd., Tokyo (Japan); Motoyama, N. [Fuji Electric Corporate Research and Development, Ltd., Kanagawa (Japan)

    1998-06-10

    Measures against Cryptosporidium parvum (C. parvum) in the waterworks are discussed. C. parvum is a pathogenic protozoan, and exists in the form of oocyst protected by a hard shell. It does not multiply in water or food, but does in human intestines and causes violent diarrhea and bellyache. A grave concern was created when many people were infected with the protozoan via tap water in Japan and the United States. Under such circumstances, ozone is used in an experiment to inactivate C. parvum. It is found that the C. parvum oocyst inactivation effect is evaluated by using a Ct value (disinfectant concentration Cmg/Ltimescontact time in minute) and that ozone treatment inactivates 90-99% of the protozoan. When various advanced water treatment technologies are being introduced for the purpose of serving safe and tasty water, the outcome of this study conveniently offers an ozone treatment method that will additionally inactivate pathogenic protozoa. Studies will be continued to elucidate the effects of factors of ozone treatment and water quality for the completion of an ideal disinfection process. Reference is made to an example of disinfection work implemented at a water purification plant of Milwaukie City, United States. 9 refs., 6 figs., 4 tabs.

  18. Molecular characterization of Danish Cryptosporidium parvum isolates

    DEFF Research Database (Denmark)

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

    2002-01-01

    The genetic polymorphism among 271 Danish Cryptosporidium isolates of human and animal origin was studied by partial amplification and sequencing of the Cryptosporidium oocyst wall protein (COWP) gene, the 18S rDNA, and a microsatellite locus.dagger Furthermore, the microsatellite locus was studi...

  19. Emissie van Cryptosporidium en Giardia door landbouwhuisdieren

    NARCIS (Netherlands)

    Schrijven JF; Bruin HAM de; Engels GB; Leenen EJTM; MGB

    1999-01-01

    In this study, the relative contributions of the pathogenic protozoa Cryptosporidium and Giardia by manure of farm animals in The Netherlands to the total yearly environmental load was studied. Manure of veal calves forms a very large source of Cryptosporidium (1.5 m 10 square 16 oocysts per year)

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

    Science.gov (United States)

    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.

  1. Epidemiology of equine Cryptosporidium and Giardia infections.

    Science.gov (United States)

    Xiao, L; Herd, R P

    1994-01-01

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

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

    Science.gov (United States)

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

    2011-04-01

    Groundwater from karst aquifers is an important source of drinking water worldwide. Outbreaks of cryptosporidiosis linked to surface water and treated public water are regularly reported. Cryptosporidium oocysts are resistant to conventional drinking water disinfectants and are a major concern for the water industry. Here, we examined conditions associated with oocyst transport along a karstic hydrosystem, and the impact of intensive exploitation on Cryptosporidium oocyst contamination of the water supply. We studied a well-characterized karstic hydrosystem composed of a sinkhole, a spring and a wellbore. Thirty-six surface water and groundwater samples were analyzed for suspended particulate matter, turbidity, electrical conductivity, and Cryptosporidium and Giardia (oo)cyst concentrations. (Oo)cysts were identified and counted by means of solid-phase cytometry (ChemScan RDI(®)), a highly sensitive method. Cryptosporidium oocysts were detected in 78% of both surface water and groundwater samples, while Giardia cysts were found in respectively 22% and 8% of surface water and groundwater samples. Mean Cryptosporidium oocyst concentrations were 29, 13 and 4/100 L at the sinkhole, spring and wellbore, respectively. Cryptosporidium oocysts were transported from the sinkhole to the spring and the wellbore, with respective release rates of 45% and 14%, suggesting that oocysts are subject to storage and remobilization in karst conduits. Principal components analysis showed that Cryptosporidium oocyst concentrations depended on variations in hydrological forcing factors. All water samples collected during intensive exploitation contained oocysts. Control of Cryptosporidium oocyst contamination during intensive exploitation is therefore necessary to ensure drinking water quality. Copyright © 2011. Published by Elsevier Ltd.

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

    Science.gov (United States)

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

    2012-06-01

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

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

    OpenAIRE

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

    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 varani...

  5. Occurence of Cryptosporidium spp. in low quality water and on vegetables in Kumasi, Ghana

    DEFF Research Database (Denmark)

    Petersen, T. B.; Petersen, H. H.; Abaidoo, R. C.

    2014-01-01

    Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp. contaminat......Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp...... of Cryptosporidium positive samples was unsuccessful, thus no conclusions can be drawn concerning sources of contamination. Nevertheless, the detection of high prevalence and concentration levels of Cryptosporidium oocysts on vegetables consumed raw and in water with direct contact to humans entails a potential risk...

  6. Sensor detection of parasite eggs and (oo-)cysts - possibilities and challenges

    DEFF Research Database (Denmark)

    Enemark, Heidi

    2015-01-01

    and cysts ((oo-)cysts) of the protozoan genera Cryptosporidium and Giardia are major causes of waterborne outbreaks of diarrhea. Methods for routine recovery and detection of waterborne Giardia and/or Cryptosporidium include filtration, immunomagnetic separation and detection by microscopy...... of immunofluorescence stained (oo-)cysts. These methods have low recovery rates, are time consuming, costly, and require well equipped laboratory facilities. Likewise, microscopy is the universal diagnostic method for detection of helminth eggs and protozoa in food and feed despite low sensitivity, difficulties...... system and ultrasound to obtain high recovery rates of apparently undamaged protozoa: 84.9% (Standard deviation (±) 4.8) for Giardia cysts and 70% (± 6.5) for Cryptosporidium oocysts. Ultrasound in the current system is tuned into a useful tool for enhanced elution of filtered (oo-)cysts. The combined...

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

    Science.gov (United States)

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

    2013-10-01

    Fifty-seven fecal samples were collected from giant pandas (Ailuropoda melanoleuca) in the China Conservation and Research Centre for the Giant Panda (CCRCGP) in Sichuan and examined for Cryptosporidium oocysts by Sheather's sugar flotation technique. An 18-year-old male giant panda was Cryptosporidium positive, with oocysts of an average size of 4.60×3.99 μm (n=50). The isolate was genetically analyzed using the partial 18S rRNA, 70 kDa heat shock protein (HSP70), Cryptosporidium oocyst wall protein (COWP) and actin genes. Multi-locus genetic characterization indicated that the present isolate was different from known Cryptosporidium species and genotypes. The closest relative was the Cryptosporidium bear genotype, with 11, 10, and 6 nucleotide differences in the 18S rRNA, HSP70, and actin genes, respectively. Significant differences were also observed in the COWP gene compared to Cryptosporidium mongoose genotype. The homology to the bear genotype at the 18S rRNA locus was 98.6%, which is comparable to that between Cryptosporidium parvum and Cryptosporidium hominis (99.2%), or between Cryptosporidium muris and Cryptosporidium andersoni (99.4%). Therefore, the Cryptosporidium in giant pandas in this study is considered as a new genotype: the Cryptosporidium giant panda genotype. © 2013 Elsevier Ireland Ltd. All rights reserved.

  8. Cryptosporidium varanii infection in leopard geckos ( Eublepharis ...

    African Journals Online (AJOL)

    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 ...

  9. Quantitative assessment of viable Cryptosporidium parvum load in commercial oysters (Crassostrea virginica) in the Chesapeake Bay.

    Science.gov (United States)

    Graczyk, Thaddeus K; Lewis, Earl J; Glass, Gregory; Dasilva, Alexandre J; Tamang, Leena; Girouard, Autumn S; Curriero, Frank C

    2007-01-01

    The epidemiological importance of increasing reports worldwide on Cryptosporidium contamination of oysters remains unknown in relation to foodborne cryptosporidiosis. Thirty market-size oysters (Crassostrea virginica), collected from each of 53 commercial harvesting sites in Chesapeake Bay, MD, were quantitatively tested in groups of six for Cryptosporidium sp. oocysts by immunofluorescent antibody (IFA). After IFA analysis, the samples were retrospectively retested for viable Cryptosporidium parvum oocysts by combined fluorescent in situ hybridization (FISH) and IFA. The mean cumulative numbers of Cryptosporidium sp. oocysts in six oysters (overall, 42.1+/-4.1) were significantly higher than in the numbers of viable C. parvum oocysts (overall, 28.0+/-2.9). Of 265 oyster groups, 221 (83.4%) contained viable C. parvum oocysts, and overall, from 10-32% (mean, 23%) of the total viable oocysts were identified in the hemolymph as distinct from gill washings. The amount of viable C. parvum oocysts was not related to oyster size or to the level of fecal coliforms at the sampling site. This study demonstrated that, although oysters are frequently contaminated with oocysts, the levels of viable oocysts may be too low to cause infection in healthy individuals. FISH assay for identification can be retrospectively applied to properly stored samples.

  10. Hyperimmune bovine colostrum treatment of moribund Leopard geckos (Eublepharis macularius) infected with Cryptosporidium sp.

    Science.gov (United States)

    Graczyk, T K; Cranfield, M R; Bostwick, E F

    1999-01-01

    Therapy based on the protective passive immunity of hyperimmune bovine colostrum (HBC) was applied to 12 moribund Leopard geckos (Eublepharis macularius) infected with Cryptosporidium sp. The geckos were lethargic and moderately to severely emaciated, weighing on average 36% of the baseline body weight value. Seven gastric HBC treatments at 1-week intervals each decreased the relative output of Cryptosporidium sp. oocysts and the prevalence of oocyst-positive fecal specimens. Histologically, after 8 weeks of therapy, seven out of 12 geckos had only single developmental stages of Cryptosporidium sp. in the intestinal epithelium, and three, one and one geckos had low, moderate and high numbers, respectively, of the pathogen developmental stages. The HBC therapy was efficacious in decreasing the parasite load in moribund geckos. Morphometric and immunologic analysis of Cryptosporidium sp. oocyst isolates originating from Leopard geckos (E. macularius) demonstrated differences between gecko-derived oocyst isolates and isolates of C. serpentis recovered from snakes.

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

    Science.gov (United States)

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

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

    Science.gov (United States)

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

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

    Science.gov (United States)

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

    2011-01-10

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

  14. Long-Term Storage of Cryptosporidium parvum for In Vitro Culture

    NARCIS (Netherlands)

    Paziewska-Harris, A.; Schoone, G.; Schallig, H. D. F. H.

    2018-01-01

    The long-term storage of Cryptosporidium life-cycle stages is a prerequisite for in vitro culture of the parasite. Cryptosporidium parvum oocysts, sporozoites, and intracellular forms inside infected host cells were stored for 6-12 mo in liquid nitrogen utilizing different cryoprotectants (dimethyl

  15. Large outbreak of Cryptosporidium hominis infection transmitted through the public water supply, Sweden.

    Science.gov (United States)

    Widerström, Micael; Schönning, Caroline; Lilja, Mikael; Lebbad, Marianne; Ljung, Thomas; Allestam, Görel; Ferm, Martin; Björkholm, Britta; Hansen, Anette; Hiltula, Jari; Långmark, Jonas; Löfdahl, Margareta; Omberg, Maria; Reuterwall, Christina; Samuelsson, Eva; Widgren, Katarina; Wallensten, Anders; Lindh, Johan

    2014-04-01

    In November 2010, ≈27,000 (≈45%) inhabitants of Östersund, Sweden, were affected by a waterborne outbreak of cryptosporidiosis. The outbreak was characterized by a rapid onset and high attack rate, especially among young and middle-aged persons. Young age, number of infected family members, amount of water consumed daily, and gluten intolerance were identified as risk factors for acquiring cryptosporidiosis. Also, chronic intestinal disease and young age were significantly associated with prolonged diarrhea. Identification of Cryptosporidium hominis subtype IbA10G2 in human and environmental samples and consistently low numbers of oocysts in drinking water confirmed insufficient reduction of parasites by the municipal water treatment plant. The current outbreak shows that use of inadequate microbial barriers at water treatment plants can have serious consequences for public health. This risk can be minimized by optimizing control of raw water quality and employing multiple barriers that remove or inactivate all groups of pathogens.

  16. Giardia spp. and Cryptosporidium spp. In the Ivaí Indigenous Land, Brazil.

    Science.gov (United States)

    Nishi, Letícia; Bergamasco, Rosângela; Toledo, Max Jean de Ornelas; Falavigna, Dina Lúcia Morais; Gomes, Mônica Lúcia; Mota, Lúcio Tadeu; Falavigna-Guilherme, Ana Lúcia

    2009-10-01

    The objective of this study was to investigate the occurrence of cysts of Giardia spp. and oocysts of Cryptosporidium spp. in waters of the Ivaí Indigenous Land, Brazil. Samples of river and spring water and of treated water were filtered and analyzed by direct immunofluorescence (Merifluor kit, Meridian Bioscience, Cincinnati, Ohio). Of 21 samples, 7 from each locality, 3 (3/7, 42.8%) from a river were positive for Giardia (mean concentration 2.57 cysts/L), and 1 (1/7, 14.3%) was positive for Cryptosporidium (6 oocysts/L). From springs, 1 sample (1/7, 14.3%) was positive for Cryptosporidium (6 oocysts/L). One sample (1/7, 14.3%) from treated water was positive for both, with 4 oocysts/L and 2 cysts/L. Giardia was the more frequent protozoan present.

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

    Science.gov (United States)

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

    2015-01-01

    The objectives of this cross-sectional study were to detect the presence of Cryptosporidium spp. and Giardia duodenalis in drinking water treatments plants (DWTPs) in Galicia (NW Spain) and to identify which species and genotype of these pathogenic protozoans are present in the water. Samples of untreated water (surface or ground water sources) and of treated drinking water (in total, 254 samples) were collected from 127 DWTPs and analysed by an immunofluorescence antibody test (IFAT) and by PCR. Considering the untreated water samples, Cryptosporidium spp. were detected in 69 samples (54.3%) by IFAT, and DNA of this parasite was detected in 57 samples (44.8%) by PCR, whereas G. duodenalis was detected in 76 samples (59.8%) by IFAT and in 56 samples (44.0%) by PCR. Considering the treated drinking water samples, Cryptosporidium spp. was detected in 52 samples (40.9%) by IFAT, and the parasite DNA was detected in 51 samples (40.1%) by PCR, whereas G. duodenalis was detected in 58 samples (45.6%) by IFAT and in 43 samples (33.8%) by PCR. The percentage viability of the (oo)cysts ranged between 90.0% and 95.0% in all samples analysed. Cryptosporidium andersoni, C. hominis, C. parvum and assemblages A-I, A-II, E of G. duodenalis were identified. The results indicate that Cryptosporidium spp. and G. duodenalis are widespread in the environment and that DWTPs are largely ineffective in reducing/inactivating these pathogens in drinking water destined for human and animal consumption in Galicia. In conclusion, the findings suggest the need for better monitoring of water quality and identification of sources of contamination. Copyright © 2014 Elsevier GmbH. All rights reserved.

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

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    A. Dellarupe

    2016-06-01

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

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

    Science.gov (United States)

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

    2016-08-01

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

  1. Elimination of viruses, bacteria and protozoan oocysts by slow sand filtration

    NARCIS (Netherlands)

    Hijnen, W.A.M.; Visser, Ate; Schijven, J.F.; Bonné, P.; Medema, Gerriet Jan

    2004-01-01

    The decimal elimination capacity (DEC) of slow sand filters (SSF) for viruses, bacteria and oocysts of Cryptosporidium has been assessed from full-scale data and pilot plant and laboratory experiments. DEC for viruses calculated from experimental data with MS2-bacteriophages in the pilot plant

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

    Science.gov (United States)

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

    2015-05-01

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

  3. [Achatina fulica Bowdich (1822) a new host of Cryptosporidium (Apicomplexa, Cryptosporidiidae) species].

    Science.gov (United States)

    Schiffler, Cinthia L; Gomes, Francimar F; Ederli, Nicole B; De Oliveira, Francisco Carlos R

    2008-09-01

    With the objective of isolate Cryptosporidium spp. in Achatina fulica s feces, 50 mollusks were collected in nine neighborhoods of the municipal of Campos dos Goytacazes, RJ to the observation of oocysts in feces. The snails were put in individuals containers and fed with water and green vegetables ad libitum until be collected a gram of feces per animal. The samples were conditioned in tubes with formalin 10% and later smear of feces were made and dyed by Ziechl-Neelsen modified technique. Of the 50 samples examined, 26 (52%) were positive for the presence of oocysts of Cryptosporidium spp. The morphology and morphometry of the oocysts showed that are a great morphologic variability. Considering the obtained results, the mollusk Achatina fulica is a host of Cryptosporidium species and can participate in the epidemic chain of the cryptosporidiosis.

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

    Directory of Open Access Journals (Sweden)

    Sotiriadou Isaia

    2013-01-01

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

  5. Prevalence of Cryptosporidium and Giardia lamblia in Water Samples from Jeddah and Makkah Cities

    Directory of Open Access Journals (Sweden)

    Haytham Ahmed Zakai

    2014-01-01

    Full Text Available Water contamination by Giardia lamblia and Cryptosporidium is one of the causes of diarrhoea throughout the world.  A total of 161 and 84 samples were collected from Jeddah and Makkah cities, respectively.  Each sample was concentrated by double centrifugation and the sediment was examined as a wet smear and after staining with Trichrome and Kinyoun stains.  The results showed that 56 (35% and 1 (0.62 % samples of Jeddah were positive for the oocyst of Cryptosporidium and cyst of Giardia, whereas only 21 (25% and 2 (2.4 % samples of Makkah showed positivity for oocysts and cyst of these parasites. Overall Cryptosporidium contamination in bottled water and water from filling stations was 6.8% and 17.4%, respectively. Maximum contamination for Cryptosporidium was recorded in tap water which was 51% and 25% in Jeddah and Makkah, respectively.

  6. Prevalence and molecular identification of Cryptosporidium isolates from pet lizards and snakes in Italy

    Directory of Open Access Journals (Sweden)

    Rinaldi L.

    2012-11-01

    Full Text Available In order to acquire prevalence and genetic data on Cryptosporidium infections in captive lizards and snakes kept as pets, a survey was conducted on 150 individual reptiles from southern Italy. Fecal samples were preserved in 5% formalin and analyzed using a commercial immunofluorescence assay (IFA for the detection of Cryptosporidium oocysts and Giardia cysts. IFA revealed the presence of Cryptosporidium oocysts in nine of the 150 samples examined (6.0%, precisely in 6/125 snakes (4.8% and in 3/25 lizards (12.0%; all fecal samples tested negative for the presence of Giardia cysts. Molecular characterization based on nested PCR amplification and sequencing of the SSU-rRNA gene, revealed the presence of Cryptosporidium serpentis in three samples from snakes (Boa constrictor constrictor, Elapheguttata guttata guttata and Python molurus.

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

    Science.gov (United States)

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

    2017-05-17

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

  8. Potential role of beavers (Castor fiber in contamination of water in the Masurian Lake District (north-eastern Poland with protozoan parasites Cryptosporidium spp. and Giardia duodenalis

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    Sroka Jacek

    2015-06-01

    Full Text Available The purpose of this study was to assess the possible influence of beavers on the contamination of lake water with zoonotic parasites Giardia duodenalis and Cryptosporidium spp., with respect to the risk to human health. A total of 79 water samples were taken around the habitats of beavers from 14 localities situated in the recreational Masurian Lake District (north-eastern Poland. Water was sampled in the spring and autumn seasons, at different distances from beavers’ lodges (0-2, 10, 30, and 50 m. The samples were examined for the presence of (oocysts of zoonotic protozoa Giardia duodenalis and Cryptosporidium spp. by direct fluorescence assay (DFA and by nested and real time PCR. By DFA, the presence of Giardia cysts was found in 36 samples (45.6% and the presence of Cryptosporidium oocysts in 26 samples (32.9%. Numbers of Giardia cysts, Cryptosporidium oocysts, and summarised (oocysts of both parasites showed a significant variation depending on locality. The numbers of Giardia cysts significantly decreased with the distance from beavers’ lodges while the numbers of Cryptosporidium oocysts did not show such dependence. The amount of Giardia cysts in samples collected in spring was approximately 3 times higher than in autumn. Conversely, a larger number of Cryptosporidium oocysts were detected in samples collected in autumn than in spring. By PCR, Giardia DNA was found in 38 samples (48.1% whereas DNA of Cryptosporidium was found in only 7 samples (8.9%. Eleven Giardia isolates were subjected to phylogenetic analysis by restriction fragment length polymorphism PCR or sequencing which evidenced their belonging to zoonotic assemblages: A (3 isolates and B (8 isolates. In conclusion, water in the vicinity of beavers’ lodges in the tested region was markedly contaminated with (oocysts of Giardia duodenalis and Cryptosporidium spp., which confirms the potential role of beavers as a reservoir of these parasites and indicates a need for

  9. Cryptosporidium Source Tracking in the Potomac River Watershed▿

    Science.gov (United States)

    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

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

    DEFF Research Database (Denmark)

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

    2007-01-01

    The performance of immunofluorescence microscopy (IF) and enzyme-linked immunosorbent assay (ELISA) in canine feces was evaluated. IF and Cryptosporidium ELISA detected 10(5) oocysts/g, while the detection limit for Giardia ELISA was 10(4) cysts/g. The Cryptosporidium ELISA showed 94% specificity...... zoonotic character of Cryptosporidium and Giardia infections in 150 asymptomatic Finnish dogs from the Helsinki area were studied. The overall proportion of dogs positive for Cryptosporidium was 5% (7/150) and that for Giardia 5% (8/150). In dogs...

  11. Importância de Cryptosporidium spp. como causa de diarréia em bezerros Importance of Cryptosporidium spp. as a cause of diarrhea in calves

    Directory of Open Access Journals (Sweden)

    Francisco L.F. Feitosa

    2008-10-01

    Full Text Available Avaliou-se a presença de oocistos de Cryptosporidium spp. em amostras de fezes de 14 bezerros e de suas mães até a oitava semana pós parição. A maior taxa de excreção de oocistos foi verificada em bezerros com sete dias de idade. Das vacas, 42,8% foram positivas para Cryptosporidium no período pós-parto. Em outra etapa deste estudo, foram acompanhados 57 bezerros positivos para Cryptosporidium, com até 30 dias de idade, provenientes de 32 propriedades leiteiras, e estudouse o grau de eliminação dos oocistos com a possível ocorrência de diarréia. Em todos os animais positivos para Cryptosporidium foi pesquisada a presença de bactérias enteropatogênicas, vírus (Rotavirus e Coronavirus e protozoários (Eimeria spp..The aim of this research was to evaluate the shedding of Cryptosporidium spp. oocysts in fecal samples from 14 calves from one dairy farm, from birth until 60 days old and from cows until eight weeks after parturition. The higher percentage of oocysts excreted was observed in 7-day-old calves. In the post-partum period 43.7% of cows were positive for Cryptosporidium oocysts. Further analyses were accomplished in 57 calves from another 32 milk farms, previously known as positive for Cryptosporidium, through oocysts fecal screening and clinical signs analyses until calves were 30 days old. Fecal samples from all animals that presented diarrhea were screened for the presence of bacteria, virus (Rotavirus and Coronavirus and protozoa (Eimeria spp..

  12. [Investigation of Cryptosporidium sp. in workers of the Van municipality slaughterhouse and in slaughtered animals].

    Science.gov (United States)

    Ciçek, Mutalip; Körkoca, Hanifi; Gül, Abdurrahman

    2008-01-01

    This study was carried out in order to investigate the prevalence of Cryptosporidium sp. in slaughtered animals and workers of the Van municipality slaughterhouse in Van. Animals slaughtered at different times and workers who had been working in different departments of the slaughter house were included in the study for three months. A total of 309 fecal specimens from animals including 167 sheep, 56 goats and 86 cattle and 87 fecal specimens from workers were examined for Cryptosporidium sp. oocysts. In slaughtered animals, the modified acid-fast staining method was used to determine the oocysts of Cryptosporidium sp. The fecal samples of slaughter workers were examined by using RIDA (R) Quick Cryptosporidium Strip Test (R-Biopharm, Germany) and the modified acid-fast staining method. Fecal samples found to be positive by stripe test were also confirmed with the ELISA method (R-Biopharm, Germany). Oocysts of Cryptosporidium sp. were found in fecal specimens of 22 sheep (13.17%), 6 goats (10.71%) and 7 cattle (8.13%). Intestinal parasites were observed in 34 fecal specimens of workers (39.08%). Cryptosporidium sp., Hymenolepis nana, Chilomastix mesnili, Endolimax nana, Iodamoeba bütschlii were found in the specimen of one worker (1.14%), Entamoeba coli in 4 workers (4.59%), Blastocystis hominis (9.19%) in 8 workers, and Giardia intestinalis (19.54%) in 17 workers.

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

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    Yousef Mirzai

    2014-04-01

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

  14. New host and distributional records for Cryptosporidium sp. (Apicomplexa: Cryptosporidiidae) from lizards (Sauria: Gekkonidae, Scincidae) from the Cook Islands and Vanuatu, South Pacific

    Science.gov (United States)

    McAllister, Chris T.; Duszynski, Donald W.; Fisher, Robert N.

    2013-01-01

    Between 1991 and 1993, 295 lizards, comprising 21 species in 2 families (Gekkonidae, Scincidae) from the Cook Islands, Fiji, Palau, Takapoto, and Vanuatu in the South Pacific, were examined for Cryptosporidium oocysts. Only 6 lizards (2%) were found to be passing Cryptosporidium oocysts in their feces, including 2 of 30 (7%) Oceania geckos, Gehyra oceanica, from Rarotonga, Cook Islands, and 4 of 26 (15%) Pacific blue-tailed skinks, Emoia caeruleocauda, from Efate Island, Vanuatu. This represents the largest survey for Cryptosporidium in Pacific island lizards, and we document 2 new host and 2 new locality records for this parasite genus.

  15. Detection of Toxoplasma gondii oocysts in water: proposition of a strategy and evaluation in Champagne-Ardenne Region, France

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    D Aubert

    2009-03-01

    Full Text Available Water is a vehicle for disseminating human and veterinary toxoplasmosis due to oocyst contamination. Several outbreaks of toxoplasmosis throughout the world have been related to contaminated drinking water. We have developed a method for the detection of Toxoplasma gondii oocysts in water and we propose a strategy for the detection of multiple waterborne parasites, including Cryptosporidium spp. and Giardia. Water samples were filtered to recover Toxoplasma oocysts and, after the detection of Cryptosporidium oocysts and Giardia cysts by immunofluorescence, as recommended by French norm procedure NF T 90-455, the samples were purified on a sucrose density gradient. Detection of Toxoplasma was based on PCR amplification and mouse inoculation to determine the presence and infectivity of recovered oocysts. After experimental seeding assays, we determined that the PCR assay was more sensitive than the bioassay. This strategy was then applied to 482 environmental water samples collected since 2001. We detected Toxoplasma DNA in 37 environmental samples (7.7%, including public drinking water; however, none of them were positive by bioassay. This strategy efficiently detects Toxoplasma oocysts in water and may be suitable as a public health sentinel method. Alternative methods can be used in conjunction with this one to determine the infectivity of parasites that were detected by molecular methods.

  16. cryptosporidium oocysts in ghanaian aids patients with diarrhoea

    African Journals Online (AJOL)

    Kateee

    or the synthetic peptide-based immunoassay (PeptiLav I and II), both obtained from Sanofi Diagnostic Pasteur, Marnes-la-. Coquette, France. Definition of diarrhoea episodes: The World Health. Organisation (WHO) criteria was used to determine diarrhoeal episodes (14). A total of three or more unformed stools in one 24-.

  17. The search for Cryptosporidium oocysts and Giardia cysts in source ...

    African Journals Online (AJOL)

    ... from any source, the risks of contamination are much greater in surface waters than in ground waters. A number of waterborne outbreaks of cryptosporidiosis, from potable and recreational water have been recorded. In many cases these outbreaks have occurred from water that complies to current microbiology standards.

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

    Science.gov (United States)

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

    2000-03-01

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

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

    Science.gov (United States)

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

    2010-12-01

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

  20. Occurrence of Cryptosporidium (Apicomplexa, Cryptosporidiidae in Crotalus durissus terrificus (Serpentes, Viperidae in Brazil

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    Karasawa Andréa Satie Matsubara

    2002-01-01

    Full Text Available The objective of the present study was to investigate the prevalence of Cryptosporidium (Apicomplexa, Cryptosporidiidae in the snake Crotalus durissus terrificus (Serpentes, Viperidae. Fifty animals were evaluated for the presence of oocysts of Cryptosporidium sp. at the time of arrival and 30 and 60 days later. Intestinal washings with saline solution (1% body weight, fecal samples, and organ scrapings were collected during the study. Oocysts were concentrated by an ether-phosphate-buffered saline sedimentation technique and then separated by a density gradient centrifugation technique. Smears were made with the sediment and submitted to modified acid-fast and auramine-rhodamine staining. Cryptosporidium-positive smears were used as controls for the experimental findings. The overall prevalence of Cryptosporidium sp. oocysts was 14%. Among the positive snakes, oocysts were detected only in the intestinal washing in two specimens, only in the feces in four specimens, and in both materials at least once in one specimen. The positive snakes were predominantly from Santa Maria da Serra city State of São Paulo (57.1%. We also observed that all of the examinations that presented positive results were obtained at least 27 days after the capture of the animals.

  1. The effects of Moringa lieifera seed powder on turbidity and sedimentation of Cryptosporidium spp. in wastewater

    DEFF Research Database (Denmark)

    Petersen, H. H.; Wolsey, I.; Dalsgaard, A.

    2013-01-01

    or water used for postharvest washing of the produce is contaminated. A laboratory study was carried out to investigate the effect of a coagulant from the seeds of Moringa oleifera (MO) in reducing Cryptosporidium parvum oocysts and turbidity in Danish wastewater. To each of five time points, 12 replicates...

  2. Avaliação da influência do formol e do hipoclorito de sódio na pesquisa de oocistos de Cryptosporidium nas fezes, através do método de heine

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    José Rafael Módolo

    1994-06-01

    Full Text Available Foram examinadas, para pesquisa de Cryptosporidium pelo método de Heine,fezes de nove bezerros com criptosporidíase, após utilização prévia de dois diferentes desinfetantes. Quanto ao formol a 10%, notou-se que não houve interferência na identificação dos oocistos, em período compreendido entre cinco minutos e 72 horas; ao ser usado o hipoclorito de sódio a 14,5%, verificou-se que depois de 30minutos os ooçistos apresentaram-se avermelhados e sem refração, dificultando o reconhecimento. Assim, recomenda-se a adição de formol a 10% à matéria fecal, conforme a etapa referida, para coibir o risco de infecção de laboratoristas pelo vírus da imunodeficiência humana (HIV, quando usada para diagnóstico atécnica mencionada.Cryptosporidium oocysts were searched by Heine 's method in stools of nine cal/s with cryptoporidiosis after stool treatment with two disinfectants, 10% paraformaldehyde solution and 14,5% sodium hypochlorite solution. After 30 minutes exposition to sodium hypochlorite solution oocysts became non refractile and acquired a reddish tinge, making their Identification difficult. No morphological alterations occured in oocysts after paraformaldehyde treatment. We recommend paraformaldehyde at 10% concentration as means of human immunodeficiency virus (HIV inactivation for routine use in stool examinations and therefore making safer those type of procedures for laboratory personnel, when using Heine 's method.

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

    Science.gov (United States)

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

    2009-12-03

    The aim of this study was to determine the prevalence of Cryptosporidium species and genotypes in birds kept in captivity in Brazil. A total of 966 samples from 18 families of birds was collected and stored in 5% potassium dichromate solution at 4 degrees C until processing. Oocysts were purified in Sheather sugar solution following extraction of genomic DNA. Molecular analyses were performed using nested-PCR for amplification of fragments of the 18S subunit of rRNA gene and of the actin gene. Amplification of Cryptosporidium DNA fragments was obtained in 47 (4.86%) samples. Sequencing of amplified fragments and phylogenetic analyses allowed the identification of Cryptosporidium baileyi in a black vulture (Coragyps atratus), a domestic chicken (Gallus gallus domesticus) and a saffron finch (Sicalis flaveola); Cryptosporidium galli in canaries (Serinus canaria), a cockatiel (Nymphicus hollandicus) and lesser seed-finches (Oryzoborus angolensis); Cryptosporidium meleagridis in a domestic chicken (G. g. domesticus); Cryptosporidium parvum in a cockatiel (N. hollandicus); Cryptosporidium avian genotype I in a canary (S. canaria) and an Indian peafowl (Pavo cristatus); Cryptosporidium avian genotype II in ostriches (Struthio camelus) and Cryptosporidium avian genotype III in a cockatiel (N. hollandicus) and a peach-faced lovebird (Agapornis roseicolis).

  4. Genotyping Cryptosporidium andersoni in cattle in Shaanxi Province, Northwestern China.

    Directory of Open Access Journals (Sweden)

    Guang-Hui Zhao

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

  5. Global modelling of Cryptosporidium in surface water

    Science.gov (United States)

    Vermeulen, Lucie; Hofstra, Nynke

    2016-04-01

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

  6. Specific detection of Neospora caninum oocysts in fecal samples from experimentally-infected dogs using the polymerase chain reaction.

    Science.gov (United States)

    Hill, D E; Liddell, S; Jenkins, M C; Dubey, J P

    2001-04-01

    Neospora caninum oocysts, passed in the feces of a definitive host (dog), were isolated, and genomic DNA was extracted. A polymerase cahin reaction (PCR) targeting the N. caninum-specific Nc 5 genomic sequence was performed using the isolated DNA. A synthesized competitor molecule containing part of the Nc 5 sequence was included in the assay as a check against false-negative PCR results and to quantify N. caninum oocyst DNA in fecal samples. A standard curve of the ratio of fluorescence intensity of PCR-amplified competitor to that of oocyst DNA was constructed to compare oocyst equivalents from fecal samples containing unknown numbers of N. caninum oocysts and to assess the sensitivity of the assay. The specificity of the assay was determined using the Nc 5-specific primers in PCR assays against other parasites likely to be found in canine feces. Genomic DNA sequences from the canine coccidians Hammondia heydorni, Cryptosporidium parvum, Sarcocystis cruzi, S. tenella, and Isospora ohioensis and the canine helminth parasites Strongyloides stercoralis, Toxocara canis, Dipylidium caninum, and Ancylostoma caninum were not amplified. In addition, genomic DNA sequences from oocysts of coccidian parasites that might contaminate dog feces, such as Hammondia hammondi, Toxoplasma gondii, or Eimeria tenella, were not amplified in the PCR assay. The assay should be useful in epidemiological surveys of both domestic and wild canine hosts and in investigations of oocyst biology in experimental infections.

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

    Science.gov (United States)

    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. Cryptosporidium parvum, a potential cause of colic adenocarcinoma

    Directory of Open Access Journals (Sweden)

    Pinon Anthony

    2007-11-01

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

  9. Identification of Cryptosporidium Species and Genotypes in Scottish Raw and Drinking Waters during a One-Year Monitoring Period▿

    Science.gov (United States)

    Nichols, R. A. B.; Connelly, L.; Sullivan, C. B.; Smith, H. V.

    2010-01-01

    We analyzed 1,042 Cryptosporidium oocyst-positive slides (456 from raw waters and 586 from drinking waters) of which 55.7% contained 1 or 2 oocysts, to determine species/genotypes present in Scottish waters. Two nested PCR-restriction fragment length polymorphism (RFLP) assays targeting different loci (1 and 2) of the hypervariable region of the 18S rRNA gene were used for species identification, and 62.4% of samples were amplified with at least one of the PCR assays. More samples (577 slides; 48.7% from raw water and 51.3% from drinking water) were amplified at locus 1 than at locus 2 (419 slides; 50.1% from raw water and 49.9% from drinking water). PCR at loci 1 and 2 amplified 45.4% and 31.7% of samples containing 1 or 2 oocysts, respectively. We detected both human-infectious and non-human-infectious species/genotype oocysts in Scottish raw and drinking waters. Cryptosporidium andersoni, Cryptosporidium parvum, and the Cryptosporidium cervine genotype (now Cryptosporidium ubiquitum) were most commonly detected in both raw and drinking waters, with C. ubiquitum being most common in drinking waters (12.5%) followed by C. parvum (4.2%) and C. andersoni (4.0%). Numerous samples (16.6% total; 18.9% from drinking water) contained mixtures of two or more species/genotypes, and we describe strategies for unraveling their identity. Repetitive analysis for discriminating mixtures proved useful, but both template concentration and PCR assay influenced outcomes. Five novel Cryptosporidium spp. (SW1 to SW5) were identified by RFLP/sequencing, and Cryptosporidium sp. SW1 was the fourth most common contaminant of Scottish drinking water (3%). PMID:20639357

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

    Science.gov (United States)

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

    2018-02-01

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

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

    Science.gov (United States)

    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

  12. Performance and oocyst shedding in broiler chickens orally infected with Cryptosporidium baileyi and Cryptosporidium meleagridis

    Czech Academy of Sciences Publication Activity Database

    Marounek, Milan; Tůmová, E.; Skřivan, M.; Pavlásek, I.; Ledvinka, Z.

    2002-01-01

    Roč. 46, - (2002), s. 203-207 ISSN 0005-2086 R&D Projects: GA ČR GA524/99/0480; GA AV ČR KSK5020115 Keywords : chickens * experimental cryptosporidiosis * performance Subject RIV: ED - Physiology Impact factor: 1.151, year: 2002

  13. A cell culture platform for Cryptosporidium that enables long-term cultivation and new tools for the systematic investigation of its biology.

    Science.gov (United States)

    Miller, Christopher N; Jossé, Lyne; Brown, Ian; Blakeman, Ben; Povey, Jane; Yiangou, Lyto; Price, Mark; Cinatl, Jindrich; Xue, Wei-Feng; Michaelis, Martin; Tsaousis, Anastasios D

    2018-03-01

    Cryptosporidium parasites are a major cause of diarrhoea that pose a particular threat to children in developing areas and immunocompromised individuals. Curative therapies and vaccines are lacking, mainly due to lack of a long-term culturing system of this parasite. Here, we show that COLO-680N cells infected with two different Cryptosporidium parvum strains produce sufficient infectious oocysts to infect subsequent cultures, showing a substantial fold increase in production, depending on the experiment, over the most optimistic HCT-8 models. Oocyst identity was confirmed using a variety of microscopic- and molecular-based methods. This culturing system will accelerate research on Cryptosporidium and the development of anti-Cryptosporidium drugs. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  14. Cryptosporidium Zoonosis in Nigeria

    African Journals Online (AJOL)

    Dr Olaleye

    Cryptosporidium Zoonosis in Nigeria. Ayinmode, A. B. and Fagbemi B. O. Department of Veterinary Microbiology and Parasitology,. Faculty of Veterinary Medicine, University of Ibadan, Ibadan, Nigeria. ABSTRACT: Cryptosporidium is a coocidian parasite that infects a wide range of vertebrate hosts including man.

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

    Science.gov (United States)

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

    2015-08-01

    Little is known of the prevalence of Cryptosporidium and Giardia in domestic cats in Western Australia and their potential role as zoonotic reservoirs for human infection. In the present study, a total of 345 faecal samples from four different sources were screened for the presence of Cryptosporidium and Giardia by PCR and genotyped by sequence analysis. Oocyst numbers and cyst numbers for Cryptosporidium and Giardia respectively were also determined using quantitative PCR assays. Cryptosporidium and Giardia were detected in 9.9% (95% CI 6.7-13.0) and 10.1% (95% CI 7.0-13.3) of cats in Western Australia respectively. Sequence analysis at the 18S rRNA locus identified five Cryptosporidium species/genotypes; C. felis (n = 8), C. muris (n = 1), C. ryanae (n = 1), Cryptosporidium rat genotype III (n = 5) and a novel genotype most closely related to Cryptosporidium rat genotype III in one isolate. This is the first report of C. ryanae and Cryptosporidium rat genotype III in cats. For Giardia, assemblage F the most commonly identified species, while only 1 assemblage sequence was detected. Since most human cases of cryptosporidiosis are caused by C. parvum and C. hominis and human cases of giardiasis are caused by G. duodenalis assemblage A and B, the domestic cats in the present study are likely to be of low zoonotic risk to pet owners in Perth. Risk analyses identified that elderly cats (more than 6 years) were more prone to Cryptosporidium and Giardia infections than kittens (less than 6 months) (P = 0.009). Clinical symptoms were not associated with the prevalence of Cryptosporidium and Giardia infections in cats. Copyright © 2015 Elsevier Inc. All rights reserved.

  16. First description of Cryptosporidium parvum in carrier pigeons (Columba livia).

    Science.gov (United States)

    Oliveira, Bruno César Miranda; Ferrari, Elis Domingos; da Cruz Panegossi, Mariele Fernanda; Nakamura, Alex Akira; Corbucci, Flávio Sader; Nagata, Walter Bertequini; Dos Santos, Bianca Martins; Gomes, Jancarlo Ferreira; Meireles, Marcelo Vasconcelos; Widmer, Giovanni; Bresciani, Katia Denise Saraiva

    2017-08-30

    The carrier pigeon and the domestic pigeon are different breeds of the species Columba livia. Carrier pigeons are used for recreational activities such as bird contests and exhibitions. Due to the close contact with humans, these birds may potentially represent a public health risk, since they can host and disseminate zoonotic parasites, such as those belonging to the genus Cryptosporidium (phylum Apicomplexa). The purpose of this work was the detection by microscopic and molecular techniques of Cryptosporidium spp. oocysts in fecal samples of carrier pigeons, and subsequently to sequence the 18S ribosomal RNA marker of positive samples to identify the species. A total of 100 fecal samples were collected individually in two pigeon breeding facilities from Formiga and Araçatuba, cities located in Minas Gerais state and São Paulo state, Brazil, respectively. The age of the birds ranged from one to 12 years; 56 were females and 44 males. Fecal smears were stained with negative malachite green, whereas the molecular characterization was based on the sequence of a ∼800bp fragment of the 18S rRNA gene. Microscopic examination of fecal smears revealed 4% (4/100) oocyst positivity. On the other hand, 7% (7/100) of positivity were found using nested PCR. Three samples were 99% to 100% similar to Cryptosporidium parvum 18S rDNA type A (Genbank AH006572) and the other three samples had 99% to 100% similarity to C. parvum 18S rDNA type B (Genbank AF308600). To our knowledge, this is the first report of C. parvum oocysts in carrier pigeons. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Evaluation of the solar water disinfection process (SODIS) against Cryptosporidium parvum using a 25-L static solar reactor fitted with a compound parabolic collector (CPC).

    Science.gov (United States)

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

    2012-02-01

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

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

    Science.gov (United States)

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

    2009-06-01

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

  19. The Association of Cryptosporidium parvum With Suspended Sediments: Implications for Transport in Surface Waters

    Science.gov (United States)

    Searcy, K. E.; Packman, A. I.; Atwill, E. R.; Harter, T.

    2003-12-01

    Understanding the transport and fate of microorganisms in surface waters is of vital concern in protecting the integrity and safety of municipal water supply systems. The human pathogen Cryptosporidium parvum is a particular public health interest, as it is ubiquitous in the surface waters of the United States, it can persist for long periods in the environment, and it is difficult to disinfect in water treatment plants. Due to its small size (5 um), low specific gravity (1.05 g/cm3), and negative surface charge, C. parvum oocysts are generally considered to move through watersheds from their source to drinking water reservoirs with little attenuation. However, the transport of the oocysts in surface waters may be mediated by interactions with suspended sediments. Batch experiments were conducted to determine the extent of C. parvum oocyst attachment to several inorganic and organic sediments under varying water chemical conditions, and settling column experiments were performed to demonstrate how these associations influence the effective settling velocity of C. parvum oocysts. Results from these experiments showed that C. parvum oocysts do associate with inorganic and organic sediments and often settle at the rate of the suspended sediment. The size and surface charge of the host suspended sediment influenced the extent of oocyst attachment as oocysts preferentially associated with particles greater than 3 um, and fewer oocysts associated with particles having a highly negative surface charge. Background water chemical conditions including ionic strength, ion composition, and pH did not have a significant effect on oocyst attachment to suspended sediments.

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

    Science.gov (United States)

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

    2010-06-01

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

  1. Natural infection with two genotypes of Cryptosporidium in red squirrels (Sciurus vulgaris) in Italy

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Hofmannová, L.; Bertolino, S.; Wauters, L.; Tosi, G.; Modrý, David

    2008-01-01

    Roč. 55, č. 2 (2008), s. 95-99 ISSN 0015-5683 R&D Projects: GA ČR GP523/07/P117; GA ČR GA524/05/0992 Institutional research plan: CEZ:AV0Z60220518 Keywords : Cryptosporidium * Sciurus vulgaris * 18S rRNA * oocyst morphology * infectivity * red squirrel Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 1.307, year: 2008

  2. Dual wavelength multiple-angle light scattering system for cryptosporidium detection

    Science.gov (United States)

    Buaprathoom, S.; Pedley, S.; Sweeney, S. J.

    2012-06-01

    A simple, dual wavelength, multiple-angle, light scattering system has been developed for detecting cryptosporidium suspended in water. Cryptosporidium is a coccidial protozoan parasite causing cryptosporidiosis; a diarrheal disease of varying severity. The parasite is transmitted by ingestion of contaminated water, particularly drinking-water, but also accidental ingestion of bathing-water, including swimming pools. It is therefore important to be able to detect these parasites quickly, so that remedial action can be taken to reduce the risk of infection. The proposed system combines multiple-angle scattering detection of a single and two wavelengths, to collect relative wavelength angle-resolved scattering phase functions from tested suspension, and multivariate data analysis techniques to obtain characterizing information of samples under investigation. The system was designed to be simple, portable and inexpensive. It employs two diode lasers (violet InGaN-based and red AlGaInP-based) as light sources and silicon photodiodes as detectors and optical components, all of which are readily available. The measured scattering patterns using the dual wavelength system showed that the relative wavelength angle-resolved scattering pattern of cryptosporidium oocysts was significantly different from other particles (e.g. polystyrene latex sphere, E.coli). The single wavelength set up was applied for cryptosporidium oocysts'size and relative refractive index measurement and differential measurement of the concentration of cryptosporidium oocysts suspended in water and mixed polystyrene latex sphere suspension. The measurement results showed good agreement with the control reference values. These results indicate that the proposed method could potentially be applied to online detection in a water quality control system.

  3. Occurrence of Cryptosporidium and Giardia in domestic animals in peri-urban communities of Kafue district, Zambia

    DEFF Research Database (Denmark)

    Siwila, J.; Phiri, I. G. K.; Enemark, Heidi L.

    2013-01-01

    Cryptosporidium spp. and Giardia duodenalis are important parasites infecting a wide range of domestic animals worldwide. The aim of the present study was to determine the occurrence of Cryptosporidium spp. and Giardia parasites in different domestic animals living in close contact with humans...... within rural/semiurban communities in Kafue district in Zambia. A single faecal sample per animal was collected from pigs, goats, dogs, ducks, chickens and pigeons and analysed by Merifluor Cryptosporidium/Giardia immunofluorescence antibody assay for the simultaneous detection of these parasites....... The faecal consistency was noted and scored as non-diarrhoeic or diarrhoeic. A total of 236 samples were collected. Cryptosporidium spp. oocysts were detected in pigs (11.5%, 17/148), goats (5.9%; 1/17), ducks (10.0%; 3/30) and chickens (14.3%; 2/14) while Giardia cysts were detected in pigs (8.1%; 12...

  4. Investigation into Cryptosporidium and Giardia in bivalve mollusks farmed in Sardinia region and destined for human consumption

    Directory of Open Access Journals (Sweden)

    Tiziana Tedde

    2013-06-01

    Full Text Available Cryptosporidium and Giardia are protozoan parasites transmitted by fecal-oral ingestion of (oocysts, and are responsible for enteritis in several animal species and humans worldwide. These (oocysts can survive for over a year in aquatic environments and can accumulate in bivalve mollusks, which filter large volumes of water. The aim of this study is to evaluate the natural occurrence of Cryptosporidium and Giardia contamination in different specimens of edible bivalves mollusks from farming sites of the western and north-eastern coasts of Sardinia. From April 2011 to February 2012, 1095 specimens of Mytilus galloprovincialis and 240 of Crassostrea gigas were sampled from Olbia and Oristano gulf and San Teodoro pond. Hepatopancreas and gills, including the labial palp, were examined for oocysts and cysts after pooling and homogenisation using different techniques: i staining for light and fluorescence microscopy; ii direct immunofluorescence (IF Merifluor® test Cryptosporidium/ Giardia (Meridian Bioscience Inc., Cincinnati, OH, USA; and iii molecular procedures. However, in the context under study, all mollusks examined with the three main diagnostic techniques were negative for both parasites pointing out the hypothetically low zoonotic risk related to Cryptosporidium and Giardia in bivalves, especially Mytilus galloprovincialis and Crassostrea gigas.

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

    Science.gov (United States)

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

    2015-05-01

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

  6. Cryptosporidium Infection in Dairy Cattle Calves and its Public Health Significance in Central Ethiopia

    Directory of Open Access Journals (Sweden)

    Manyazewal Anberber Zeleke

    2017-05-01

    Full Text Available Cryptosporidium spp. are common intestinal protozoan parasites that causes diarrhoea in neonates and young calves. This longitudinal study was conducted at two large dairy cattle farms in central Ethiopia during February/2014 to June/2015 to determine the age-related distribution of Cryptosporidium species, to identify risk factors of the disease and to assess the public health significance of the parasite. Thirty calves born to these dairy farms were followed-up from birth to three months of age, and 270 faecal samples were collected and examined by the Modified Ziehl-Neelsen, PCR-RFLP and Sequencing. Cryptosporidium was detected from week 1 to 3 months of age with an overall prevalence of 14.8%, Peak of the infection was at two weeks of age when 12 of the 30 calves (40% shedded oocysts. Cryptosporidium parvum and C. andersoni were identified in pre-weaned and post-weaned calves, respectively. Phylogenetic analysis showed clustering of the C. parvum isolates from this study with GenBank sequences for C. parvum bovine genotype IIa and IId subtypes. This study showed the predominance of the zoonotic C. parvum species in pre-weaned calves and demonstrated that this age group of calves pose the greatest risk for human infection. Due attention on the management of pre-weaned calves is recommended to prevent transmission of the infection to humans and lessen contamination of the environment by oocysts.

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

    DEFF Research Database (Denmark)

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

    2013-01-01

    Pigs are a potential source of contamination with Cryptosporidium spp., which can lead to infection in humans. Two species C. parvum and C. hominis can cause an acute diarrheal illness in humans, which can become severe in e.g. patients with HIV. The oocyst can survive for long periods in the env......Pigs are a potential source of contamination with Cryptosporidium spp., which can lead to infection in humans. Two species C. parvum and C. hominis can cause an acute diarrheal illness in humans, which can become severe in e.g. patients with HIV. The oocyst can survive for long periods...... in the environment and is resistant to many common disinfectants. In order to estimate the prevalence of the zoonotic parasite Cryptosporidium in organic pigs and to improve our knowledge of the parasite epidemiology, the prevalence was monitored four times between September 2011 and June 2012 in three Danish...... organic pig farms. Faecal samples were collected for examination of Cryptosporidium spp. with a total of 994 pigs grouped as sows, fatteners, young pigs and piglets. The number of pigs in each age group was 298, 232, 315 and 161 respectively, distributed on the three farms. Faecal samples were collected...

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

    DEFF Research Database (Denmark)

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

    2007-01-01

    The genetic diversity of Cryptosporidium spp. and Giardia duodenalis from dairy cattle and pigs in Denmark was determined in the present study. Faecal samples from 1237 pigs and 1150 cattle originating from 50 sow herds and 50 dairy herds, respectively, were analysed for the presence of the two...... parasites by immunofluorescence microscopy. A large proportion of the (oo)cyst containing samples were selected for molecular characterization. Sequencing and phylogenetic analysis of the 18S rDNA locus and/or the HSP70 gene of 183 pig and 154 cattle isolates of Cryptosporidium revealed the presence of C....... suis, pig genotype II, C. parvum (cattle genotype), C. bovis, Cryptosporidium deer-like genotype and a novel C. suis-like genotype. For both cattle and pigs, a host age-related change in distribution of species/genotypes was observed. The zoonotic C. parvum (cattle genotype) was most prevalent in young...

  9. Polymerase chain reaction and nested-PCR approaches for detecting Cryptosporidium in water catchments of water treatment plants in Curitiba, State of Paraná, Brazil

    Directory of Open Access Journals (Sweden)

    Silvia Cristina Osaki

    2013-06-01

    Full Text Available Introduction Cryptosporidium is an important protozoan cause of waterborne disease worldwide of concern to public health authorities. To prevent outbreaks of cryptosporidiosis, the monitoring of this parasite in drinking water is necessary. In the present work, the polymerase chain reaction (PCR and nested-PCR techniques were used to detect Cryptosporidium in raw water from catchment points of four water treatment plants (WTP in Curitiba, Paraná, Brazil. Methods First, DNA extraction techniques were tested in samples containing decreasing amount of oocysts in reagent water, and PCR and nested-PCR with specific primers for 18SSU rDNA of Cryptosporidium were conducted to determine their sensitivity. In reagent water, a commercial extraction kit provided the best analytical sensitivity, and PCR and nested-PCR allowed the detection of five and two oocysts, respectively, with the primers XIAOR/XIAOF and XIAO1F/XIAO2R. Results In the spiking experiments, only the PCR with the primers AWA995F/AWA1206R was successful at detecting concentrations of 0.1 oocysts/mL. Two catchments samples of raw water and/or water sludge from four WTPs were contaminated with Cryptosporidium. Conclusions The application of the techniques to monitor Cryptosporidium in water and detect contamination in water catchments of WTPs in Curitiba are discussed in the present work.

  10. [Isolation and identification of cow-origin Cryptosporidium isolates in Hefei].

    Science.gov (United States)

    Sun, Tao; Liu, Wei; Wang, Ju-Hua; Xue, Xiu-Heng; Zhao, Chang-Cheng; Li, Pei-Ying

    2011-12-01

    To isolate cow-origin Cryptosporidium in Hefei, and identify its species. 285 dairy cattle fecal samples collected from a farm in Hefei were examined by using floating saturated solution of sucrose and modified acid-fast staining. Cryptosporidium oocysts were isolated and purified from positive fecal samples. Genetic DNA was extracted to be the template. According to the sequence of 18S rRNA gene and HSP70 gene from Cryptosporidium sp., the primers were designed and synthesized. The PCR products were amplified by PCR and nested-PCR. The nested PCR products were cloned and sequenced. Homology searches and phylogenic tree construction were done by DNAStar software. Five fecal samples were positive by morphological methods with an infection rate of 1.8% (5/285). Oocysts from the 5 positive fecal samples were elliptical or ovoid detected by using floating saturated solution of sucrose and modified acid-fast staining with the size of 7.37 microm x 6.13 microm and 7.58 microm x 6.20 microm, and a shape index of 1.20 and 1.22, respectively. Nested-PCR resulted in a 18S rRNA and HSP70 gene fragments with approximately 250 bp and 325 bp, respectively. The five isolates showed a high level of nucleic acid identity with sequence data of the 18S rRNA gene of Cryptosporidium andersoni (DQ989573), and they were clustered in the same clade. The highest HSP70 gene sequence identity was found among the five isolates and other reported C. andersoni isolates (AY954892 and DQ989576), and they were placed into the same clade. The cow-origin Cryptosporidium isolates derived from Hefei is Cryptosporidium andersoni.

  11. Cryptosporidium sp. in children suffering from acute diarrhea at Uberlândia City, State of Minas Gerais, Brazil

    Directory of Open Access Journals (Sweden)

    Margareth Leitão Gennari-Cardoso

    1996-10-01

    Full Text Available This study's objective was to search for Cryptosporidium sp. in diarrheic feces from children aged zero to 12 years and cared for at medical units within Universidade Federal de Uberlândia or at a private practice in Uberlândia, State of Minas Gerais, Brazil, from September 1992 to August 1993. Three fecal samples preserved in 10% formalin, were collected from 94 children. Oocyst concentration was performed through Ritchie's (modified method and staining of fecal smears for each sample (total of 1128 slides was done by the "Safranin/Methylene Blue" and the "Kinyoun (modified" techniques. The Hoffmann, Pons & Janer method was also employed to look for other enteroparasites. From 94 children, 4.26% excreted fecal Cryptosporidium oocysts. The infection seemed to vary according to age: 5.08% of patients aged zero to two years old; 33.33% of those aging eight to ten years (P>0.05. Cryptosporidium appeared in November, December and March, during the rainy season. 20.21% of the children harbored at least one enteroparasite different from Cryptosporidium, mainly Giardia intestinalis (12.77%. From Cryptosporidium infected patients, two had only this kind, another harbored Giardia intestinalis; the last one hosted Strongyloides stercoralis.

  12. Cryptosporidium and Giardia in different age groups of Danish cattle and pigs - Occurrence and management associated risk factors

    DEFF Research Database (Denmark)

    Maddox-Hyttel, Charlotte; Langkjær, Rikke Breinhold; Enemark, Heidi L.

    2006-01-01

    groups. Risk factors were evaluated by using proportional odds models with (oo)cyst excretion levels divided into four categories as response. Among the numerous risk factors examined, only a few were demonstrated to have a statistically significant influence, e.g. the use of an empty period in the calf....../calves 1-12 months. The faecal samples were purified and the number of (oo)cysts quantified. The study revealed an age-specific herd prevalence of Cryptosporidium of 16, 31 and 100% for sows, piglets and weaners, respectively, and of 14, 96 and 84% for cows, young calves and older calves, respectively...

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

    Science.gov (United States)

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

    2015-06-01

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

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

    Science.gov (United States)

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

    2012-01-01

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

  15. Cryptosporidium: A Guide to Water Filters

    Science.gov (United States)

    ... Tap Water Many but not all available home water filters remove Cryptosporidium . Some filter designs are more suitable for removal of Cryptosporidium than others. Filters that have the words "reverse osmosis" on the label protect against Cryptosporidium . Some other ...

  16. Cryptosporidium sp. in lizards

    Czech Academy of Sciences Publication Activity Database

    Koudela, Břetislav; Modrý, D.

    1998-01-01

    Roč. 45, č. 1 (1998), s. 8 ISSN 1066-5234. [Cryptosporidium sp. in lazards. 01.01.1998-02.01.1998, Praha] R&D Projects: GA ČR GA508/95/0273; GA AV ČR IPP2020702 Subject RIV: fp - Other Medical Disciplines

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

    Science.gov (United States)

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

    2011-12-01

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

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

    Directory of Open Access Journals (Sweden)

    Xunde Li

    2015-12-01

    Full Text Available 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.

  19. Modelling the impact of sanitation, population growth and urbanization on human emissions of Cryptosporidium to surface waters—a case study for Bangladesh and India

    Science.gov (United States)

    Vermeulen, Lucie C.; de Kraker, Jelske; Hofstra, Nynke; Kroeze, Carolien; Medema, Gertjan

    2015-09-01

    Cryptosporidium is a protozoan parasite that can cause diarrhoea. Human faeces are an important source of Cryptosporidium in surface waters. We present a model to study the impact of sanitation, urbanization and population growth on human emissions of Cryptosporidium to surface waters. We build on a global model by Hofstra et al (2013 Sci. Total Environ. 442 10-9) and zoom into Bangladesh and India as illustrative case studies. The model is most sensitive to changes in oocyst excretion and infection rate, and to assumptions on the share of faeces reaching the surface water for different sanitation types. We find urban centres to be hotspots of human Cryptosporidium emissions. We estimate that 53% (Bangladesh) and 91% (India) of total emissions come from urban areas. 50% of oocysts come from only 8% (Bangladesh) and 3% (India) of the country area. In the future, population growth and urbanization may further deteriorate water quality in Bangladesh and India, despite improved sanitation. Under our ‘business as usual’ (‘sanitation improvements’) scenario, oocyst emissions will increase by a factor 2.0 (1.2) for India and 2.9 (1.1) for Bangladesh between 2010 and 2050. Population growth, urbanization and sanitation development are important processes to consider for large scale water quality modelling.

  20. Reduction of Cryptosporidium, Giardia, and Fecal Indicators by Bardenpho Wastewater Treatment.

    Science.gov (United States)

    Schmitz, Bradley W; Moriyama, Hitoha; Haramoto, Eiji; Kitajima, Masaaki; Sherchan, Samendra; Gerba, Charles P; Pepper, Ian L

    2018-06-19

    Increased demand for water reuse and reclamation accentuates the importance for optimal wastewater treatment to limit protozoa in effluents. Two wastewater treatment plants utilizing advanced Bardenpho were investigated over a 12-month period to determine the incidence and reduction of Cryptosporidium, Giardia, Cyclospora, and fecal indicators. Results were compared to facilities that previously operated in the same geographical area. Protozoa (oo)cysts were concentrated using an electronegative filter and subsequently detected by fluorescent microscopy and/or PCR methods. Cryptosporidium and Giardia were frequently detected in raw sewage, but Cyclospora was not detected in any wastewater samples. Facilities with Bardenpho treatment exhibited higher removals of (oo)cysts than facilities utilizing activated sludge or trickling filters. This was likely due to Bardenpho systems having increased solid wasting rates; however, this mechanism cannot be confirmed as sludge samples were not analyzed. Use of dissolved-air-flotation instead of sedimentation tanks did not result in more efficient removal of (oo)cysts. Concentrations of protozoa were compared with each other, Escherichia coli, somatic coliphage, and viruses (pepper mild mottle virus, Aichi virus 1, adenovirus, and polyomaviruses JC and BK). Although significant correlations were rare, somatic coliphage showed the highest potential as an indicator for the abundance of protozoa in wastewaters.

  1. Comparative efficacy of curcumin and paromomycin against Cryptosporidium parvum infection in a BALB/c model.

    Science.gov (United States)

    Asadpour, Mohammad; Namazi, Fatemeh; Razavi, Seyed Mostafa; Nazifi, Saeed

    2018-01-30

    Cryptosporidium is a ubiquitous protozoan parasite causing gastrointestinal disorder in various hosts worldwide. The disease is self-limiting in the immunocompetent but life-threatening in immunodeficient individuals. Investigations to find an effective drug for the complete elimination of the Cryptosporidium infection are ongoing and urgently needed. The current study was undertaken to examine the anti-cryptosporidial efficacy of curcumin in experimentally infected mice compared with that of paromomycin. Oocysts were isolated from a pre-weaned dairy calf and identified as Cryptosporidium parvum using a nested- polymerase chain reaction (PCR) on Small subunit ribosomal ribonucleic acid (SSU rRNA) gene and sequencing analysis. One hundred and ten female BALB/c mice were divided into five groups. Group 1 was infected and treated with curcumin; Group 2 infected and treated with paromomycin; Group 3 infected without treatment; Group 4 included uninfected mice treated with curcumin, and Group 5 included uninfected mice treated with distilled water for 11 successive days, starting on the first day of oocyst shedding. The oocyst shedding was recorded daily. At days 0, 3, 7, and 11 of post treatments, five mice from each group were killed humanly; jejunum and ileum tissue samples were processed for histopathological evaluation and counting of oocyst on villi, simultaneously. Furthermore, total antioxidant capacity (TAC) and malondialdehyde (MDA) concentrations in affected tissues were also measured in different groups. By treatments, tissue lesions and the number of oocyst on villi of both jejunum and ileum were decreased with a time-dependent manner. In comparison with Group 3, oocyst shedding was stopped at the end of treatment period in both groups 1 and 2 without recurrence at 10days after drug withdrawal. Also, TAC was increased and the MDA concentrations were decreased in Group 1. Moreover, paromomycin showed acceptable treatment outcomes during experiment and its

  2. Virulence of geographically different Cryptosporidium parvum isolates in experimental animal model

    Science.gov (United States)

    Sayed, Fatma G.; Hamza, Amany I.; Galal, Lamia A.; Sayed, Douaa M.; Gaber, Mona

    2016-10-01

    Cryptosporidium parvum is a coccidian parasite which causes gastrointestinal disease in humans and a variety of other mammalian species. 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 study aimed to investigate infectivity and virulence of two Cryptosporidium parvum “Iowa isolate” (CpI) and a “local water isolate” (CpW). Thirty-three Swiss albino mice have been divided into three groups: Negative control Group (C), the CpI group infected with “Iowa isolate “and the CpW group infected with C. parvum oocysts isolated from a local water supply. Infectivity and virulence have been measured by evaluating clinical, parasitological and histological aspects of infection. Significant differences were detected regarding oocysts shedding rate, clinical outcomes, and the histopathological picture of the intestine, lung, and brain. It was concluded that the local water isolate is significantly more virulent than the exported one.

  3. Relationships between the occurrence of Giardia and Cryptosporidium and physicochemical properties of marine waters of the Pacific Coast of Mexico.

    Science.gov (United States)

    Magana-Ordorica, Dalia; Mena, Kristina; Valdez-Torres, Jose B; Soto-Beltran, Marcela; Leon-Felix, Josefina; Chaidez, Cristobal

    2010-12-01

    Untreated sewage has adversely affected the quality of marine recreational waters worldwide. Exposure to marine recreational water with poor microbial quality may pose a threat to bathers. The objectives of this study were to assess the effect of physicochemical parameters on Cryptosporidium and Giardia presence in marine recreational water of Sinaloa, Mexico, by Logistic Regression Analyses. Thirty-two 10-litre water samples were collected from two tourist beaches, Altata and Mazatlan, between November 2006 and May 2007. Water samples were processed by the EPA 1623 method and pH, temperature, salinity and turbidity were also determined. Cryptosporidium and Giardia were present in 71 and 57% of the samples collected from Altata, respectively. In Mazatlan, Cryptosporidium and Giardia were found in 83 and 72% of the samples, respectively. The overall concentration of Cryptosporidium ranged from 150 to 2,050 oocysts/10 L with an average of 581 oocysts/10 L and Giardia ranged from 10 to 300 cysts/10 L with an average of 73 cysts/10 L. The occurrence of both parasites increased in water with decreasing temperatures and increasing turbidity of the water.

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

    Directory of Open Access Journals (Sweden)

    Roberta Flávia Ribeiro Rolando

    2012-06-01

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

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

    Science.gov (United States)

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

    2017-01-01

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

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

    OpenAIRE

    Agnes Kurniawan; Sri W. Dwintasari; Herbowo A. Soetomenggolo; Septelia I. Wanandi

    2009-01-01

    Aim To identify the frequency of Cryptosporidium infection in children below 3 years old by examining concentrated long term preserved stool using PCR detection of 18S rRNA gene and compared with modified acid fast staining technique.Methods Hundred eighty eight stools from children ≤ 3 years old were stored for 13 months in 2.5% K2Cr2O7 solution at 40C. Cryptosporidium oocysts were isolated by water-ether concentration technique. The concentrates were smeared onto object glass and stained wi...

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

    Science.gov (United States)

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

    2013-11-15

    Infection by Cryptosporidium serpentis is one of the most important diseases in reptiles and is characterized by chronic clinical or subclinical infection and the presence of hypertrophic gastritis, food regurgitation, progressive weight loss, mortality, and intermittent or continuous shedding of oocysts in the feces. The objectives of this study were to standardize an indirect enzyme-linked immunosorbent assay (ELISA) to detect antibodies against C. serpentis and to evaluate the clinical, parasitological, and humoral immune response in snakes naturally infected with C. serpentis. Twenty-one snakes naturally infected with C. serpentis and housed at the Butantan Institute, São Paulo, Brazil, underwent clinical and parasitological analyses for C. serpentis infection through daily records of clinical signs and a monthly survey of fecal shedding of oocysts using the Kinyoun's acid-fast staining. The serological evaluation was performed monthly by indirect ELISA using crude total antigen from oocysts of C. serpentis to detect anti-C. serpentis antibodies. Clinical symptoms consisted of food regurgitation, inappetence, and progressive weight loss. The parasitological analysis revealed intermittent fecal shedding of a variable number of oocysts in all snakes, with positivity in 85.32% (157/184) of the samples. The indirect ELISA was positive in 68.25% (86/126) of the samples. A humoral immune response was observed in most animals; however, fluctuating antibodies levels, leading to alternating positive and negative results, were observed in most snakes. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. Cryptosporidium spp. and Giardia sp. in aquatic mammals in northern and northeastern Brazil.

    Science.gov (United States)

    Borges, João Carlos; Lima, Danielle Dos; da Silva, Edson Moura; Moreira, André Lucas de Oliveira; Marmontel, Miriam; Carvalho, Vitor Luz; Amaral, Rodrigo de; Lazzarini, Stella Maris; Alves, Leucio Câmara

    2017-09-20

    Cryptosporidium and Giardia are protozoans that can infect humans and wild and domestic animals. Due to the growing importance of diseases caused by protozoan parasites in aquatic species, we aimed to evaluate the frequency of infection by Cryptosporidium spp. and Giardia sp. in aquatic and marine mammals in the northern and northeastern regions of Brazil. We collected 553 fecal samples from 15 species of wild-ranging and captive aquatic mammals in northern and northeastern Brazil. All samples were analyzed by the Kinyoun technique for identification of Cryptosporidium spp. oocysts. Giardia sp. cysts were identified by means of the centrifugal-flotation technique in zinc sulfate solution. Subsequently, all samples were submitted for direct immunofluorescence testing. The overall frequency of infection was 15.55% (86/553) for Cryptosporidium spp. and 9.04% (50/553) for Giardia sp. The presence of Cryptosporidium spp. was detected in samples from 5 species: neotropical river otter Lontra longicaudis (15.28%), giant otter Pteronura brasiliensis (41.66%), Guiana dolphin Sotalia guianensis (9.67%), Amazonian manatee Trichechus inunguis (16.03%), and Antillean manatee T. manatus (13.79%). Giardia sp. was identified in L. longicaudis (9.23%), P. brasiliensis (29.16%), pygmy sperm whale Kogia breviceps (100%), dwarf sperm whale K. sima (25%), S. guianensis (9.67%), T. inunguis (3.81%), and T. manatus (10.34%). This is the first report of Cryptosporidium spp. in L. longicaudis, P. brasiliensis, and S. guianensis, while the occurrence of Giardia sp., in addition to the 2 otter species, was also identified in manatees, thus extending the number of hosts susceptible to these parasitic agents.

  9. Antibody responses to a Cryptosporidium parvum rCP15/60 vaccine

    OpenAIRE

    Alexandra J. Burton; Daryl V. Nydam; Gary Jones; Jennifer Zambriski; Thomas C. Linden; Graham Cox; Randy Davis; Alicia Brown; Dwight D. Bowman

    2009-01-01

    Cryptosporidium parvum is a zoonotic apicomplexa-protozoan pathogen that causes gastroenteritis and diarrhoea in mammals worldwide. The organism is transmitted by ingestion of oocysts, which are shed in faeces, and completes its lifecycle in a single host.^1^ C. parvum is ubiquitous on dairy operations worldwide and is one of the leading causes of diarrhoea in calves on these farms.^2,3^ Here, for the first time, we describe the antibody response in a large group of cows to a recombinant C. p...

  10. Prevalence of Cryptosporidium spp. and other intestinal parasites in children with diarrhea

    Directory of Open Access Journals (Sweden)

    Mutalip Çiçek

    2011-03-01

    Full Text Available This study was planned to determine the role of Cryptosporidium sp. and other intestinal parasites in the diarrheal diseases in children with 0-15 years old Van district.Materials and methods: In this study, stool samples of 450 children were examined for parasites. In the study, nativ-lugol, formaldehyde-ethyl acetate sedimentation methods and trichrome staining methods were used to detect parasites in stool samples. Additionally, sedimentation methods and modified acid fast staining method were used to detect the Cryptosporidium oocysts.Results: Parasites were found in 154 (34.2% among 450 children’s with diarrhea. In this study; the ratios of parasites were as follow: Giardia intestinalis 13.5%, Blastocystis hominis 10%, Entamoeba coli 3.78%, Cryptosporidium spp. 2.2%, Hymenolepis nana 1.33 %ve Ascaris lumbricoides 1.11%.Entamoeba histolytica/Entamoeba dispar 0.89%, Chilomastix mesnili 1.78%, Iodamoeba butschlii 0.89%, Entamoeba hartmanni 0.89%, Trichomonas hominis 0.67%, Enteromonas hominis 0.67%,Conclusion: In the investigate, it was found that Giardia intestinalis and Blastocystis hominis were most prominent agents in children with diarrhea in our vicinity and Cryptosporidium spp also was an important agent which should be investigated carefully in especially risk group in routine laboratory studies.

  11. Novel Cryptosporidium bat genotypes III and IV in bats from the USA and Czech Republic.

    Science.gov (United States)

    Kváč, Martin; Hořická, Anna; Sak, Bohumil; Prediger, Jitka; Salát, Jiří; Širmarová, Jana; Bartonička, Tomáš; Clark, Mark; Chelladurai, Jeba Rose Jennifer Jesudoss; Gillam, Erin; McEvoy, John

    2015-10-01

    Bats from the families Rhinolophidae (n = 90) and Vespertilionidae (n = 191) in the USA and Czech Republic were screened for the presence of Cryptosporidium by microscopic and molecular analysis of faecal samples collected from rectum of dissected animals and from the ground beneath roosting sites. Cryptosporidium oocysts were not detected in any of the 281 faecal specimens examined using the aniline-carbol-methyl violet staining method. Nested PCR amplification, sequencing and phylogenetic analysis of the small ribosomal subunit rRNA and actin genes were used to identify isolates and infer evolutionary relationships. Cryptosporidium parvum was identified in a western small-footed bat (Myotis ciliolabrum) from the USA and a common pipistrelle bats (Pipistrellus pipistrellus) from the Czech Republic. Two novel genotypes were identified and named Cryptosporidium bat genotype III and IV. Bat genotype III was found in two big brown bats (Eptesicus fuscus) from the USA. Bat genotype IV was detected in two common pipistrelle bats from the Czech Republic.

  12. Mechanics of the Toxoplasma gondii oocyst wall

    Science.gov (United States)

    The ability of microorganisms to survive under extreme conditions is closely related to the physicochemical properties of their wall. In the ubiquitous protozoan parasite Toxoplasma gondii, the oocyst stage possesses a bilayered wall that protects the dormant but potentially infective parasites from...

  13. Método rápido para la observación de Cryptosporidium en heces Rapid method for detection of cryptosporidium in stools

    Directory of Open Access Journals (Sweden)

    Graciela Barona

    1993-02-01

    Full Text Available Entre agosto de 1990 y diciembre de 1991 se examinaron 120 muestras de materia fecal de niños o adultos que consultaron por diarrea, sugestiva de ser causada por Cryptosporidium spp. En todos los casos se realizó la coloración con Lugol-Nigroslna, que proponemos, y se hizo la confirmación con la de Ziehl Neelsen modificada, pese a su limitación de teñir con el mismo patrón de coloración el Cryptosporldium y estructuras diferentes a él. En 20 (16.6% muestras (12 de niños y 8 de adultos se identificaron ooquistes de Cryptosporldlum spp y todas se confirmaron como positivas por la coloración de Ziehl Neelsen modificada. Dado que no siempre es fácil la observación de parásitos de poca prevalencia sugerimos esta coloración como ensayo de rutina porque ayuda a distinguir los ooquistes de Cryptosporidium y mejora la observación de todos los protozoarios.

    We examined 120 stool specimens from patients with diarrheal disease, suspected of being infected with Cryptosporidium. Preliminary observation was made with a Lugol-Nigrosine stain and confirmation with modified Ziehl-Neelsen. Twenty specimens (12 from children and 8 from adults (16.6% were positive for Cryptosporidium oocysts andevery one of them was confirmed with ZN stain. Since It may be difficult to detect low-prevalence parasites we suggest routine use of Lugol-Nigrosine which is useful for the detection of Cryptosporidium as well as of other protozoa.

  14. Molecular characterization of Cryptosporidium spp. in calves (Bos taurus and Bos indicus in the Formiga city, Minas Gerais - Brazil

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    Roberto César Araujo Lima

    2014-02-01

    Full Text Available Cryptosporidiosis is a waterborne disease, has as aggravating the difficulty of preventing environmental contamination and lack of effective therapeutic measures. With marked importance to the cattle, causes inflammation and intestinal villous atrophy resulting in loss of absorptive surface. This study aimed to perform molecular characterization of Cryptosporidium spp. in calves in the city of Formiga, Minas Gerais. A total of 300 faeces samples from Holstein calves, Nelore and indefinite breed, both healthy, were evaluated by negative contrast staining technique of malachite green and through the reaction of nested PCR for amplification of DNA fragments of the 18S subunit of the RNA gene ribosomal. Occurrence of 5.33 % ( 16/300 for malachite green and 4.66 % ( 14/300 by PCR was observed, whereas no correlation was found between positive and variables studied. Through molecular characterization were identified Cryptosporidium andersoni and Cryptosporidium ryanae species. In conclusion, we observed a low incidence of infection and elimination of Cryptosporidium spp. oocysts, the absence of clinical signs in animals, strong agreement between the results obtained by the two techniques. Beyond, with the molecular characterization ( nested PCR , species of C. andersoni and C. ryanae were diagnosed in age groups not present in the literature. These two species of Cryptosporidium are described above for the first time parasitizing cattle in the state of Minas Gerais.

  15. Molecular characterisation of Cryptosporidium spp. in lambs in the South Central region of the State of São Paulo

    Directory of Open Access Journals (Sweden)

    A.S. Zucatto

    2015-04-01

    Full Text Available Considering the proximity of sheep farmers to animals that are possibly diseased or releasing fecal oocysts into the environment and the marked pathogenicity in lambs, the aim of this study was to determine the occurrence and to molecularly characterize the infection by Cryptosporidium spp. in lambs in the South Central region of the state of São Paulo, Brazil. A total of 193 fecal samples were collected from sheep of several breeds, males and females, aged up to one year. Polymerase chain reaction (nested-PCR was used to amplify DNA fragments from the subunit 18S rRNA gene and indicated 15% positivity; sequencing of amplified fragments was possible for 19 samples. Analysis of the obtained sequences showed that the identified species were Cryptosporidium xiaoi for 15 samples, constituting thus the first molecular characterization study of this Cryptosporidium species in Brazil. Cryptosporidium ubiquitum was identified for three samples and Cryptosporidium meleagridis for one sample; the latter two are considered zoonotic species.

  16. Acción de distintos coagulantes para la eliminación de Cryptosporidium spp. en el proceso de potabilización del agua The action of different coagulants to remove Cryptosporidium during the process of water treatment

    Directory of Open Access Journals (Sweden)

    B. Abramovich

    2004-06-01

    Full Text Available Cryptosporidium es uno de los microorganismos de mayor interés desde el punto de vista de la Salud Pública y constituye un problema prioritario para las plantas y organismos reguladores de agua. Debido a su pequeño tamaño y a su resistencia a la cloración, la eliminación por el proceso de potabilización es una tarea compleja. En este trabajo se analizó la efectividad de distintos coagulantes utilizados comúnmente en tal proceso para lograr la remoción de los ooquistes. Se trabajó con la prueba de jarras (Jar Test. Se halló que: 1 Los coagulantes con agregado de polímeros coadyuvantes producen remociones de ooquistes superiores a 2 log. 2 Un valor bajo de turbiedad no asegura una remoción óptima de los parásitos. 3 La adición de polielectrolitos al cloruro férrico disminuye la variabilidad tanto en la turbiedad final como en la eliminación de Cryptosporidium.Cryptosporidium is one of the microorganisms of main concern from the point of view of Public Health, being a priority problem for water treatment plants and water regulatory institutions. Due to its small size and resistance to chlorination, Cryptosporidium removal during the process of drinking water treatmentis a hard task. The effectiveness of different coagulants commonly used in the process of removal of oocysts was analyzed. Thetechnique used was the Jar Test. It was found that: 1 coagulants with the addition of polimeric coadjuvants produce over 2 logs of oocyst removal; 2 a low value in turbidity does not necessarily mean optimal parasite removal, and 3 the addition of polyelectrolites to ferric chloride diminishes variability, both in final turbidity and Cryptosporidium removal.

  17. Effect of gamma-irradiation on oocysts of Eimeria necatrix

    International Nuclear Information System (INIS)

    Singh, J.; Gill, B.S.

    1975-01-01

    Effect of γ radiation on oocysts of Eimeria necatrix was investigated. It was observed that oocysts exposed to 200kR or above did not sporulate. Irradiation at 10 to 150kR caused a progressive decrease in sporulation. Irradiation affected normal development of unsporulated oocysts as the zygote protoplasm divided into unequal masses or was shattered into granules. Increase in the intensity of irradiation of sporulated oocysts resulted in the progressive decrease in severity of the resultant infections in chicks and their effects - mortality, type of lesions developed, total oocyst production and immunity produced - were comparable with infections induced by decreasing the number of unirradiated oocysts. Infection produced by 1000 unirradiated oocysts was comparable with that resulting from 50,000 oocysts irradiated at 25kR. Infection obtained with 20,000 unexposed oocysts approximated to that produced by 50,000 oocysts irradiated at 2.5kR. It was concluded that irradiation abolished infectivity of the oocysts/sporozoites rather than bringing about attenuation of the parasite. (author)

  18. Identification and molecular characterization of Cryptosporidium and Giardia in children and cattle populations from the province of Alava, North of Spain

    Energy Technology Data Exchange (ETDEWEB)

    Cardona, Guillermo A. [Livestock Laboratory, Regional Government of Alava, Ctra. de Azua 4, 01520 Vitoria-Gasteiz (Spain); Carabin, Helene [Department of Biostatistics and Epidemiology, College of Public Health, Oklahoma University Health Sciences Center, 801 Northeast 13th Street, Room 309, Oklahoma City, OK 73104 (United States); Goni, Pilar [Department of Microbiology, Preventive Medicine and Public Health, Faculty of Medicine, University of Zaragoza, Domingo Miral s/n, 50009 Zaragoza (Spain); Arriola, Larraitz [Epidemiology Unit, Public Health Division of Guipuzcoa, Basque Government, Av. Navarra 4, 2013 San Sebastian (Spain); Robinson, Guy [UK Cryptosporidium Reference Unit, Public Health Wales, Microbiology ABM, Swansea, Singleton Hospital, Swansea SA2 8QA (United Kingdom); Fernandez-Crespo, Juan C. [Sub-direction of Public Health of Alava, Department of Health, Basque Government, Avda. Santiago 11, 01002 Vitoria-Gasteiz (Spain); Clavel, Antonio [Department of Microbiology, Preventive Medicine and Public Health, Faculty of Medicine, University of Zaragoza, Domingo Miral s/n, 50009 Zaragoza (Spain); Chalmers, Rachel M. [UK Cryptosporidium Reference Unit, Public Health Wales, Microbiology ABM, Swansea, Singleton Hospital, Swansea SA2 8QA (United Kingdom); Carmena, David, E-mail: d.carmena@imperial.ac.uk [MRC Clinical Sciences Centre, Faculty of Medicine, Imperial College, Hammersmith Hospital Campus, Du Cane Road, London W12 0NN (United Kingdom)

    2011-12-15

    The prevalence of and factors associated with the protozoan enteropathogens Cryptosporidium and Giardia have been investigated in selected children and cattle populations from the province of Alava (Northern Spain). The presence of these organisms was detected in fecal samples using commercially available coproantigen-ELISA (CpAg-ELISA) and immunochromatographic (ICT) assays. A total of 327 caregivers of children participants were asked to answer questions on risk factors potentially associated to the prevalence of Cryptosporidium and Giardia, including water-use practices, water sports and contact with domestic or pet animals. Molecular analyses were conducted using a nested-PCR technique to amplify the small-subunit (SSU) rRNA gene of Cryptosporidium and the triosephosphate isomerase (tpi) gene of Giardia. Cryptosporidium oocysts and Giardia cysts were found in 3 and 16 samples using the CpAg-ELISA, and in 5 and 9 samples using the ICT test, respectively. Cryptosporidium and Giardia were also found in 7 and 17 samples by CpAg-ELISA, and 4 and 14 samples by ICT, respectively, of 227 cattle fecal samples. The overall Cryptosporidium and Giardia infection prevalences, based on a Bayesian approach accounting for the imperfect sensitivities and specificities of both diagnostic tests, were estimated to 1.0% (95% BCI: 0.2%-2.8%) and 3.1% (1.5%-5.3%) in children and 3.0% (0.5%-9.2%) and 1.4% (0.0%-6.4%) in cattle, respectively. In humans, a single Cryptosporidium isolate was characterized as C. hominis. Of seven Giardia isolates, four were identified as assemblage B, two as assemblage A-II and one was a mixed assemblage B + A-II infection. No Cryptosporidium or Giardia isolates could be obtained from cattle samples. Although limited, these results seem to suggest that cattle are unlikely to be an important reservoir of zoonotic Cryptosporidium and/or Giardia in the province of Alava.

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

    DEFF Research Database (Denmark)

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

    2015-01-01

    Although pigs are commonly infected with Cryptosporidium spp. and Giardia duodenalis, including potentially zoonotic species or genotypes, little is known about age-related infection levels, seasonal differences and genetic variation in naturally infected pigs raised in organic management systems....... Therefore, the current study was conducted to assess seasonal and age-related variations in prevalence and infection intensity of Cryptosporidium and Giardia, evaluate zoonotic potential and uncover correlations between species/genotypes, infection intensity and faecal consistency. Shedding of oocysts...... and cysts ((oo-) cysts) was monitored at quarterly intervals (September 2011 to June 2012) in piglets (n=152), starter pigs (n=234), fatteners (n=230) and sows (n=240) from three organic farms in Denmark. (Oo-) cysts were quantified by immunofluorescence microscopy; and 56/75 subsamples from Cryptosporidium...

  20. Assessment of the capacity of slow sand filtration to eliminate Cryptosporidium oocysts

    NARCIS (Netherlands)

    Hijnen, W.A.M.; Dullemont, Y.J.; Bosklopper, K.T.G.J.; Schijven, J.F.; Medema, Gerriet Jan

    2006-01-01

    Decimal Elimination Capacity (DEC) of the slow sand filters of the Dutch drinking water Companies was assessed; first by literature review, followed by evaluation of the removal of environmental spores of sulphite-reducing clostridia (SSRC) and small-sized centric diatoms (SSCD) as surrogates.

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

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    João Carlos Gomes Borges

    2009-03-01

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

  2. Evaluation of fenbendazole for treatment of Giardia infection in cats concurrently infected with Cryptosporidium parvum.

    Science.gov (United States)

    Keith, Carey L; Radecki, Steven V; Lappin, Michael R

    2003-08-01

    To determine whether fenbendazole effectively eliminates Giardia organisms from chronically infected cats that have a concurrent Cryptosporidium parvum infection. 16 clinically normal cats. Eight cats with chronic concurrent Giardia and C parvum infections received fenbendazole (50 mg/kg, PO, q 24 h) for 5 days (treatment-group cats). Feces from each cat were collected and processed 3 days weekly for 23 days after treatment. By use of an immunofluorescent assay for detection of Giardia lamblia cysts and C parvum oocysts, organism numbers were counted and scored. Fecal results from treatment-group cats were compared with those of 8 untreated cats with Giardia infection but no C parvum infection (control-group cats). Four of 8 treatment-group cats had consistently negative results for Giardia infection after treatment. These 4 cats had consistently positive results for C parvum oocysts prior to treatment and consistently negative results after treatment. One treatment-group cat had positive results for cysts on all fecal samples, and 3 treatment-group cats had 1 to 3 negative results and then resumed shedding large numbers of cysts; each of these cats had consistently positive results for C parvum oocysts. When compared with control-group cats, treatment-group cats shed less Giardia cysts during week 1 after treatment but not during week 2. Administration of fenbendazole decreases Giardia cyst shedding to less than detectable numbers in some cats. In our study, persistent C parvum infection may have been associated with failure of fenbendazole to eliminate Giardia infection.

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

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    Raquel Saucier Gomes

    2009-10-01

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

  4. Coagulation, flocculation, dissolved air flotation and filtration in the removal of Giardia spp. and Cryptosporidium spp. from water supply.

    Science.gov (United States)

    Andreoli, Fernando César; Sabogal-Paz, Lyda Patricia

    2017-11-15

    Removing protozoa from a water supply using coagulation, flocculation, dissolved air flotation (DAF) and filtration on a bench scale was evaluated. Calcium carbonate flocculation with and without immunomagnetic separation (IMS) was chosen to detect Giardia spp. cysts and Cryptosporidium spp. oocysts in the studied samples. The results indicated that DAF removed between 1.31 log and 1.79 log of cysts and between 1.08 log and 1.42 log of oocysts. The performance was lower in filtration, with the removal of 1.07 log-1.44 log for cysts and 0.82 log-0.98 log for oocysts. The coagulation, flocculation, DAF and filtration steps removed more than 2.2 log of cysts and oocysts from the water studied. However, protozoa were detected in the filtered water, even with turbidity values of 0.2 NTU. The recovery of the detection method met the international criteria and was higher when there was no IMS. Including the third acid dissociation in the IMS was critical to improve the performance of the protocol tested. However, there was an increase in the technical and analytical complexity and costs. It was also observed that the efficiency of the treatment was linked to the performance of the selected method of detecting protozoa.

  5. A study on protozoan infections (Giardia, Entamoeba, Isoapora and Cryptosporidium in stray dogs in Ilam province

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    S Kakekhani

    2011-11-01

    Full Text Available Giardia, Entamoeba, Isospora and Cryptosporidium are important protozoan parastites that caused diarrhea in human and animals. In the present study, fecal samples were collected fresh, directly from the rectum of 112 stray dogs in Ilam province. Giardia and Entamoeba were concentrated by using the formalin ether sedimentation method followed by the trichrome and iodine staining technique andCryptosporidium  oocysts  were  concentrated  by  using  the  formalin  ether  sedimentation  method  followed by the modified Ziehl-Neelsen staining technique. Of 112 stray dogs, protozoan infections were detected from feces of 46 dogs (41.07% that Giardia infection was detected from feces of 21 dogs (18.75%, Isospora 17 (15.17%, Cryptosporidium 8 (7.14% and synchronization infection to 2 protozoan in 9 dogs (8.03% and to 3 protozoan in 3 (2.67%. In the present study not observed to Entamoeba. No statistically significant differences in prevalence of protozoan parasites occurred between female (34.21 % and male (55.5 % stray dogs (p>0/05. But statistically significant differences in prevalence occurred between 1≥0 and 0 ≥1 stray dogs (p>0/05. So that stray dogs of Ilam province can cause infection of human water and food sources.

  6. Disposable swim diaper retention of Cryptosporidium-sized particles on human subjects in a recreational water setting.

    Science.gov (United States)

    Amburgey, James E; Anderson, J Brian

    2011-12-01

    Cryptosporidium is a chlorine-resistant protozoan parasite responsible for the majority of waterborne disease outbreaks in recreational water venues in the USA. Swim diapers are commonly used by diaper-aged children participating in aquatic activities. This research was intended to evaluate disposable swim diapers for retaining 5-μm diameter polystyrene microspheres, which were used as non-infectious surrogates for Cryptosporidium oocysts. A hot tub recirculating water without a filter was used for this research. The microsphere concentration in the water was monitored at regular intervals following introduction of microspheres inside of a swim diaper while a human subject undertook normal swim/play activities. Microsphere concentrations in the bulk water showed that the majority (50-97%) of Cryptosporidium-sized particles were released from the swim diaper within 1 to 5 min regardless of the swim diaper type or configuration. After only 10 min of play, 77-100% of the microspheres had been released from all swim diapers tested. This research suggests that the swim diapers commonly used by diaper-aged children in swimming pools and other aquatic activities are of limited value in retaining Cryptosporidium-sized particles. Improved swim diaper solutions are necessary to efficiently retain pathogens and effectively safeguard public health in recreational water venues.

  7. Labeling surface epitopes to identify Cryptosporidium life stages using a scanning electron microscopy-based immunogold approach.

    Science.gov (United States)

    Edwards, Hanna; Thompson, R C Andrew; Koh, Wan H; Clode, Peta L

    2012-02-01

    The Apicomplexan parasite Cryptosporidium parvum is responsible for the widespread disease cryptosporidiosis, in both humans and livestock. The nature of C. parvum infection is far from understood and many questions remain in regard to host-parasite interactions, limiting successful treatment of the disease. To definitively identify a range of C. parvum stages in cell culture and to begin to investigate host cell interactions in some of the lesser known life stages, we have utilized a combined scanning electron microscopy and immunolabeling approach, correlating high resolution microstructural information with definitive immunogold labeling of Cryptosporidium stages. Several life cycle stages, including oocysts, merozoites I, trophozoites, gamonts and microgametocytes, were successfully immunolabeled in an in vitro model system. Developing oocysts were clearly immunolabeled, but this did not persist once excystation had occurred. Immunolabeling visualized on the host cell surface adjacent to invasive merozoites is likely to be indicative of receptor shedding, with merozoites also initiating host responses that manifested as abnormal microvilli on the host cell surface. Small sub-micron stages such as microgametocytes, which were impossible to identify as single entities without immunolabeling, were readily visualized and observed to attach to host cells via novel membranous projections. Epicellular parasites also expressed Cryptosporidium-derived epitopes within their encapsulating membrane. These data have allowed us to confidently identify a variety of C. parvum stages in cell culture at high resolution. With this, we provide new insight into C. parvum - host cell interactions and highlight future opportunities for investigating and targeting receptor-mediated interactions between Cryptosporidium life cycle stages and host cells. Copyright © 2011 Elsevier Ltd. All rights reserved.

  8. Cryptosporidium Priming Is More Effective than Vaccine for Protection against Cryptosporidiosis in a Murine Protein Malnutrition Model

    Science.gov (United States)

    Bartelt, Luther A.; Bolick, David T.; Kolling, Glynis L.; Zaenker, Edna I.; Lara, Ana M.; Noronha, Francisco Jose; Cowardin, Carrie A.; Moore, John H.; Turner, Jerrold R.; Warren, Cirle A.; Buck, Gregory A.; Guerrant, Richard L.

    2016-01-01

    Cryptosporidium is a major cause of severe diarrhea, especially in malnourished children. Using a murine model of C. parvum oocyst challenge that recapitulates clinical features of severe cryptosporidiosis during malnutrition, we interrogated the effect of protein malnutrition (PM) on primary and secondary responses to C. parvum challenge, and tested the differential ability of mucosal priming strategies to overcome the PM-induced susceptibility. We determined that while PM fundamentally alters systemic and mucosal primary immune responses to Cryptosporidium, priming with C. parvum (106 oocysts) provides robust protective immunity against re-challenge despite ongoing PM. C. parvum priming restores mucosal Th1-type effectors (CD3+CD8+CD103+ T-cells) and cytokines (IFNγ, and IL12p40) that otherwise decrease with ongoing PM. Vaccination strategies with Cryptosporidium antigens expressed in the S. Typhi vector 908htr, however, do not enhance Th1-type responses to C. parvum challenge during PM, even though vaccination strongly boosts immunity in challenged fully nourished hosts. Remote non-specific exposures to the attenuated S. Typhi vector alone or the TLR9 agonist CpG ODN-1668 can partially attenuate C. parvum severity during PM, but neither as effectively as viable C. parvum priming. We conclude that although PM interferes with basal and vaccine-boosted immune responses to C. parvum, sustained reductions in disease severity are possible through mucosal activators of host defenses, and specifically C. parvum priming can elicit impressively robust Th1-type protective immunity despite ongoing protein malnutrition. These findings add insight into potential correlates of Cryptosporidium immunity and future vaccine strategies in malnourished children. PMID:27467505

  9. Checking the detail in retail: Occurrence of Cryptosporidium and Giardia on vegetables sold across different counters in Chandigarh, India.

    Science.gov (United States)

    Utaaker, Kjersti Selstad; Kumar, Anil; Joshi, Himanshu; Chaudhary, Suman; Robertson, Lucy J

    2017-12-18

    Fresh produce has been recognized as a vehicle of infection for protozoan parasites, particularly Cryptosporidium, and, to a lesser extent, Giardia. For both parasites, outbreaks associated with fresh produce have been documented. Although documented outbreaks tend to be from industrialized countries, contamination of fresh produce with these parasites is a global issue. In developing countries, infections with these parasites are often endemic in the community, and basic infrastructure and hygiene measures may be inadequate, thus the likelihood of contamination of fresh produce with these parasites may be higher. Realization of the importance of this transmission route comes against a backdrop of raw salads and more Western culinary habits gaining a foothold, and fresh produce being encouraged as part of the diet due to their associated health benefits. However, if consumption of uncooked fresh produce is going to increase its market sector in India, it is important that it is safe. In this study, various types of fresh produce obtained from three types of vendors in Chandigarh, a major city in Northern India, were analyzed for contamination with Cryptosporidium oocysts and Giardia cysts using a method that has been previously validated in inter-laboratory spiking experiments. A total of 284 samples of different fresh produce items were analyzed, obtained from the different retailers situated in different societal layers of the city. The overall prevalence of contamination of fresh produce with these parasites was just under 11%, with 6% of the vegetables contaminated with Cryptosporidium oocysts, and 5% with Giardia cysts. Contaminated vegetables included turnip, cabbage, carrot, chili, coriander, cucumber, radishes, and tomatoes. Molecular analyses identified contamination with Cryptosporidium parvum and Giardia duodenalis of Assemblage A and Assemblage D, indicating that contamination from animals may be of relevance. Although the prevalence of contamination is

  10. Motility precedes egress of malaria parasites from oocysts

    Science.gov (United States)

    Klug, Dennis; Frischknecht, Friedrich

    2017-01-01

    Malaria is transmitted when an infected Anopheles mosquito deposits Plasmodium sporozoites in the skin during a bite. Sporozoites are formed within oocysts at the mosquito midgut wall and are released into the hemolymph, from where they invade the salivary glands and are subsequently transmitted to the vertebrate host. We found that a thrombospondin-repeat containing sporozoite-specific protein named thrombospondin-releated protein 1 (TRP1) is important for oocyst egress and salivary gland invasion, and hence for the transmission of malaria. We imaged the release of sporozoites from oocysts in situ, which was preceded by active motility. Parasites lacking TRP1 failed to migrate within oocysts and did not egress, suggesting that TRP1 is a vital component of the events that precede intra-oocyst motility and subsequently sporozoite egress and salivary gland invasion. DOI: http://dx.doi.org/10.7554/eLife.19157.001 PMID:28115054

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

    Directory of Open Access Journals (Sweden)

    Patrícia de Lucca

    2009-12-01

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

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

    Directory of Open Access Journals (Sweden)

    Karine Thivierge

    2016-04-01

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

  13. Prevalence and molecular characterization of Cryptosporidium species in cattle and buffalo calves in Mumbai region of India.

    Science.gov (United States)

    Hingole, A C; Gudewar, J G; Pednekar, R P; Gatne, M L

    2017-03-01

    Faecal samples of cattle and buffaloes of Mumbai region collected between November 2012 to June 2013 were analysed by conventional and molecular tools to note the prevalence of cryptosporidiosis and species involved in the infection. Conventional analysis viz., direct faecal smear examination, faecal smear examination after normal saline sedimentation, Sheather's floatation and Sheather's floatation sedimentation smear methods demonstrated oocysts of Cryptosporidium in 141 (36.06 %) of 391 samples with higher occurrence in buffaloes (36.99 %) than cattle (34.48 %). Diarrhoeic loose faeces showed higher prevalence (42.07 %) than apparently normal faeces (31.72 %) irrespective of the host species. When data were arranged as per age groups viz., calves of 0-1 month, 1-2 months, 2-3 months and adults, the highest prevalence was noted in the youngest group (47.12 %) declining gradually with the advancing age with lowest (6.25 %) in adults indicating inverse correlation between prevalence rate and age of the host. These differences were statistically significant in case of buffaloes. Cryptosporidium andersoni was tentatively identified by morphometric analysis. By employing molecular tools like nested PCR, PCR-RFLP and sequence analysis of few samples showed good correlation in the identification of species of Cryptosporidium involved in the infection and demonstrated occurrence of C. parvum , C. ryanae and C. bovis. Thus all the four commonly occurring bovine species of Cryptosporidium were encountered in the study area which appears to be a first record reporting the occurrence of Cryptosporidium with species level identification in large ruminants from Western region of India. Additionally, the public health significance of C. parvum was also discussed in light of epidemiological factors pertaining to the region.

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

    Directory of Open Access Journals (Sweden)

    Agnes Kurniawan

    2009-09-01

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

  15. Prevalence of Cryptosporidium species and Giardia intestinalis ...

    African Journals Online (AJOL)

    Cryptosporidium species and Giardia intestinalis cause diarrheal infections in humans and other vertebrate animals globally and are considered to be of great public health importance. The study was conducted to determine the prevalence Cryptosporidium species and G. intestinalis infections among patients attending ...

  16. Occurrence of Cryptosporidium species coproantigens on a ...

    African Journals Online (AJOL)

    This study was carried out to assess the potential of animals, used for teaching and research, as a source of Cryptosporidium infection for students and staff of a University in Nigeria. Faecal samples from 185 animals reared on the teaching and research farm were collected and examined for Cryptosporidium spp. antigens ...

  17. Cryptosporidium Infections Among Children in Peru

    Centers for Disease Control (CDC) Podcasts

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

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

    Directory of Open Access Journals (Sweden)

    H. E. Laubach

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

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

    Science.gov (United States)

    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.

  20. Differing susceptibilities of Eimeria acervulina, Eimeria maxima, and Eimeria tenella oocysts to desiccation.

    Science.gov (United States)

    Jenkins, Mark C; Parker, Carolyn; O'Brien, Celia; Miska, Katarzyna; Fetterer, Raymond

    2013-10-01

    Outbreaks of avian coccidiosis may occur when susceptible chickens are raised on litter containing viable Eimeria oocysts. The purpose of this study was to compare the relative sensitivities of Eimeria acervulina, Eimeria maxima, and Eimeria tenella oocysts to dessication. Sporulated E. acervulina, E. maxima, or E. tenella oocysts were incorporated into gelatin beads and incubated at 32 C for 0, 1, 2, or 3 days. In vitro oocyst excystation rates were measured for each combination of Eimeria species and incubation time. Day-old broiler chicks were allowed to ingest the oocysts-containing beads, and total oocyst production was measured from days 5-8 post-inoculation. Although no effect on excystation was observed, E. maxima oocysts displayed greater resistance to drying compared to E. acervulina and E. tenella oocysts. Eimeria acervulina oocyst production decreased 100-fold after 1-2 days incubation. Eimeria tenella oocysts were slightly more resistant to drying in that a 100-fold decrease in oocyst production was delayed until 2 days. For both E. acervulina and E. tenella , very few oocysts were observed after 3 days incubation. Eimeria maxima oocyst production remained high at all time points. Subsequent studies revealed E. maxima oocyst production was ablated only after 5 days incubation. These findings may explain in part the observed prevalence of E. maxima in litter from commercial poultry operations.

  1. First molecular characterization of Cryptosporidium in Yemen.

    Science.gov (United States)

    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.

  2. Occurrence of Giardia and Cryptosporidium in captive chimpanzees (Pan troglodytes), mandrills (Mandrillus sphinx) and wild Zanzibar red colobus monkeys (Procolobus kirkii).

    Science.gov (United States)

    Debenham, John J; Atencia, Rebeca; Midtgaard, Fred; Robertson, Lucy J

    2015-04-01

    The aim of this study was to investigate the occurrence of Giardia duodenalis and Cryptosporidium spp. in primates and determine their zoonotic or anthropozoonotic potential. Direct immunofluorescence was used to identify Giardia and Cryptosporidium from faecal samples. PCR and DNA sequencing was performed on positive results. Giardia cysts were identified from 5.5% (5/90) of captive chimpanzees and 0% (0/11) of captive mandrills in the Republic of Congo; 0% (0/10) of captive chimpanzees in Norway; and 0% of faecal samples (n = 49) from wild Zanzibar red colobus monkeys. Two Giardia positive samples were also positive on PCR, and sequencing revealed identical isolates of Assemblage B. Cryptosporidium oocysts were not detected in any of the samples. In these primate groups, in which interactions with humans and human environments are quite substantial, Giardia and Cryptosporidium are rare pathogens. In chimpanzees, Giardia may have a zoonotic or anthropozoonotic potential. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  3. Infection by Cryptosporidium parvum in renal patients submitted to renal transplant or hemodialysis

    Directory of Open Access Journals (Sweden)

    Chieffi Pedro Paulo

    1998-01-01

    Full Text Available The frequency of infection by Cryptosporidium parvum was determined in two groups of renal patients submitted to immunosuppression. One group consisted of 23 renal transplanted individuals, and the other consisted of 32 patients with chronic renal insufficiency, periodically submitted to hemodialysis. A third group of 27 patients with systemic arterial hypertension, not immunosuppressed, was used as control. During a period of 18 months all the patients were submitted to faecal examination to detect C. parvum oocysts, for a total of 1 to 6 tests per patient. The results showed frequencies of C. parvum infection of 34.8%, 25% and 17.4%, respectively, for the renal transplanted group, the patients submitted to hemodialysis and the control group. Statistical analysis showed no significant differences among the three groups even though the frequency of C. parvum infection was higher in the transplanted group. However, when the number of fecal samples containing C. parvum oocysts was taken in account, a significantly higher frequency was found in the renal transplanted group.

  4. Detection and Resolution of Cryptosporidium Species and Species Mixtures by Genus-Specific Nested PCR-Restriction Fragment Length Polymorphism Analysis, Direct Sequencing, and Cloning ▿

    Science.gov (United States)

    Ruecker, Norma J.; Hoffman, Rebecca M.; Chalmers, Rachel M.; Neumann, Norman F.

    2011-01-01

    Molecular methods incorporating nested PCR-restriction fragment length polymorphism (RFLP) analysis of the 18S rRNA gene of Cryptosporidium species were validated to assess performance based on limit of detection (LoD) and for detecting and resolving mixtures of species and genotypes within a single sample. The 95% LoD was determined for seven species (Cryptosporidium hominis, C. parvum, C. felis, C. meleagridis, C. ubiquitum, C. muris, and C. andersoni) and ranged from 7 to 11 plasmid template copies with overlapping 95% confidence limits. The LoD values for genomic DNA from oocysts on microscope slides were 7 and 10 template copies for C. andersoni and C. parvum, respectively. The repetitive nested PCR-RFLP slide protocol had an LoD of 4 oocysts per slide. When templates of two species were mixed in equal ratios in the nested PCR-RFLP reaction mixture, there was no amplification bias toward one species over another. At high ratios of template mixtures (>1:10), there was a reduction or loss of detection of the less abundant species by RFLP analysis, most likely due to heteroduplex formation in the later cycles of the PCR. Replicate nested PCR was successful at resolving many mixtures of Cryptosporidium at template concentrations near or below the LoD. The cloning of nested PCR products resulted in 17% of the cloned sequences being recombinants of the two original templates. Limiting-dilution nested PCR followed by the sequencing of PCR products resulted in no sequence anomalies, suggesting that this method is an effective and accurate way to study the species diversity of Cryptosporidium, particularly for environmental water samples, in which mixtures of parasites are common. PMID:21498746

  5. Identification and molecular characterization of Cryptosporidium and Giardia in children and cattle populations from the province of Álava, North of Spain

    International Nuclear Information System (INIS)

    Cardona, Guillermo A.; Carabin, Hélène; Goñi, Pilar; Arriola, Larraitz; Robinson, Guy; Fernández-Crespo, Juan C.; Clavel, Antonio; Chalmers, Rachel M.; Carmena, David

    2011-01-01

    The prevalence of and factors associated with the protozoan enteropathogens Cryptosporidium and Giardia have been investigated in selected children and cattle populations from the province of Álava (Northern Spain). The presence of these organisms was detected in fecal samples using commercially available coproantigen-ELISA (CpAg-ELISA) and immunochromatographic (ICT) assays. A total of 327 caregivers of children participants were asked to answer questions on risk factors potentially associated to the prevalence of Cryptosporidium and Giardia, including water-use practices, water sports and contact with domestic or pet animals. Molecular analyses were conducted using a nested-PCR technique to amplify the small-subunit (SSU) rRNA gene of Cryptosporidium and the triosephosphate isomerase (tpi) gene of Giardia. Cryptosporidium oocysts and Giardia cysts were found in 3 and 16 samples using the CpAg-ELISA, and in 5 and 9 samples using the ICT test, respectively. Cryptosporidium and Giardia were also found in 7 and 17 samples by CpAg-ELISA, and 4 and 14 samples by ICT, respectively, of 227 cattle fecal samples. The overall Cryptosporidium and Giardia infection prevalences, based on a Bayesian approach accounting for the imperfect sensitivities and specificities of both diagnostic tests, were estimated to 1.0% (95% BCI: 0.2%–2.8%) and 3.1% (1.5%–5.3%) in children and 3.0% (0.5%–9.2%) and 1.4% (0.0%–6.4%) in cattle, respectively. In humans, a single Cryptosporidium isolate was characterized as C. hominis. Of seven Giardia isolates, four were identified as assemblage B, two as assemblage A-II and one was a mixed assemblage B + A-II infection. No Cryptosporidium or Giardia isolates could be obtained from cattle samples. Although limited, these results seem to suggest that cattle are unlikely to be an important reservoir of zoonotic Cryptosporidium and/or Giardia in the province of Álava.

  6. Characterization and separation of Cryptosporidium and Giardia cells using on-chip dielectrophoresis.

    Science.gov (United States)

    Narayanan Unni, Harikrishnan; Hartono, Deny; Yue Lanry Yung, Lin; Mah-Lee Ng, Mary; Pueh Lee, Heow; Cheong Khoo, Boo; Lim, Kian-Meng

    2012-03-01

    Dielectrophoresis (DEP) has been shown to have significant potential for the characterization of cells and could become an efficient tool for rapid identification and assessment of microorganisms. The present work is focused on the trapping, characterization, and separation of two species of Cryptosporidium (C. parvum and C. muris) and Giardia lambia (G. lambia) using a microfluidic experimental setup. Cryptosporidium oocysts, which are 2-4 μm in size and nearly spherical in shape, are used for the preliminary stage of prototype development and testing. G. lambia cysts are 8-12 μm in size. In order to facilitate effective trapping, simulations were performed to study the effects of buffer conductivity and applied voltage on the flow and cell transport inside the DEP chip. Microscopic experiments were performed using the fabricated device and the real part of Clausius-Mossotti factor of the cells was estimated from critical voltages for particle trapping at the electrodes under steady fluid flow. The dielectric properties of the cell compartments (cytoplasm and membrane) were calculated based on a single shell model of the cells. The separation of C. muris and G. lambia is achieved successfully at a frequency of 10 MHz and a voltage of 3 Vpp (peak to peak voltage).

  7. Cryptosporidium infections in Denmark, 2010-2014

    DEFF Research Database (Denmark)

    Stensvold, Christen Rune; Ethelberg, Steen; Hansen, Louise

    2015-01-01

    . RESULTS: A total of 689 Cryptosporidium-positive stool samples were submitted by 387 patients. Limiting case episodes to two months (60 days), a total of 388 case episodes representing 387 patients were identified. Cryptosporidiosis was most common among infants and toddlers. Moreover, a peak in incidence...... was observed among younger adults aged 23-24 years. In 43 Cryptosporidium-positive faecal samples, identification was performed to species and subtype level. Cryptosporidium parvum was found in 34 samples, C. hominis in eight, and C. meleagridis in one sample; C. parvum subtypes IIaA15G2R1 (n = 10) and IIaA16G...

  8. Effect of halofuginone lactate on the occurrence of Cryptosporidium parvum and growth of neonatal dairy calves.

    Science.gov (United States)

    Jarvie, B D; Trotz-Williams, L A; McKnight, D R; Leslie, K E; Wallace, M M; Todd, C G; Sharpe, P H; Peregrine, A S

    2005-05-01

    Thirty-one Holstein bull calves were purchased at birth from 3 dairy farms in Eastern Ontario. Each calf was assigned at random to oral treatment with either 5 mg of halofuginone lactate in 10.0 mL of aqueous carrier solution (Halocur, base comprised 10 mg of benzoic acid, 100 mg of lactic acid, and 0.3 mg of tartrazine) or 10 mL of placebo (Halocur base minus the active ingredient, halofuginone lactate) administered 15 to 30 min after morning milk feeding for the first 7 d of life. Intakes of milk, calf starter, and water, and fecal consistency score were recorded daily for 56 d. Calf weights were recorded weekly for 56 d. Fecal samples were taken from all calves at approximately 2, 7, 14, 21, and 28 d of age for isolation of Cryptosporidium parvum oocysts. Logistic and linear regression analyses were used to assess the effect of treatment on the incidence of diarrhea and C. parvum infection status. The odds of C. parvum shedding among calves in the halofuginone lactate-treated group was 70% lower than the odds of shedding among calves in the placebo group. In calves treated with halofuginone lactate, no oocyst shedding occurred until 2 wk of age, whereas 12.5% of calves in the placebo group began shedding oocysts during wk 1. From all ages of placebo-treated calves, 31 of 73 samples (42.5%) were positive for C. parvum, whereas only 15 of 67 samples (22.4%) from all ages of halofuginone lactate-treated calves tested positive. The largest number of C. parvum-positive samples occurred in the third week of life. There was a significant delay of 3.1 d in the incidence of diarrhea among calves treated with halofuginone lactate. Intake of milk and starter, body weight gains, and age at weaning were not significantly different between treatment groups.

  9. Oocyst wall formation and composition in coccidian parasites

    Directory of Open Access Journals (Sweden)

    Kelly Mai

    2009-03-01

    Full Text Available The oocyst wall of coccidian parasites is a robust structure that is resistant to a variety of environmental and chemical insults. This resilience allows oocysts to survive for long periods, facilitating transmission from host to host. The wall is bilayered and is formed by the sequential release of the contents of two specialized organelles - wall forming body 1 and wall forming body 2 - found in the macrogametocyte stage of Coccidia. The oocyst wall is over 90% protein but few of these proteins have been studied. One group is cysteine-rich and may be presumed to crosslink via disulphide bridges, though this is yet to be investigated. Another group of wall proteins is rich in tyrosine. These proteins, which range in size from 8-31 kDa, are derived from larger precursors of 56 and 82 kDa found in the wall forming bodies. Proteases may catalyze processing of the precursors into tyrosine-rich peptides, which are then oxidatively crosslinked in a reaction catalyzed by peroxidases. In support of this hypothesis, the oocyst wall has high levels of dityrosine bonds. These dityrosine crosslinked proteins may provide a structural matrix for assembly of the oocyst wall and contribute to its resilience.

  10. Effect of irradiation (gamma rays) on the biology of Eimeria tenella oocysts

    Energy Technology Data Exchange (ETDEWEB)

    Bajwa, R.S.; Gill, B.S.

    1977-01-01

    Effect of gamma rays on the biology of the progeny of the irradiated Eimeria tenella oocysts was investigated. The parent inoculum of sporulated oocysts was exposed to 5 to 60 kR (gamma rays). These oocysts were fed to chicks. The oocysts voided by the chicks were collected and sporulated. The sporulation rate, pathogenicity, immunogenicity and reproduction potential of these oocysts--the progeny of the irradiated oocysts--were compared with those of the unirradiated oocysts. It was observed that increase of irradiation dose caused progressive decrease in the pathogenicity of the oocyst suspension. The oocysts exposed to 30 and 40 kR produced only mild infections whereas those exposed to 50 kR and above were noninfective. No difference in pathogenicity, immunogenicity and reproduction potential of unirradiated oocysts and the oocysts progeny of the irradiated oocysts was seen. It was concluded, therefore, that the effect of irradiation was limited to the inoculum exposed to it, and was not transmissible to the progeny of the irradiated oocysts.

  11. Cryptosporidium en Giardia in Nederlandse zwembaden

    NARCIS (Netherlands)

    Schets FM; Engels GB; Leenen EJTM; MGB

    2003-01-01

    Zwembad gerelateerde explosies van cryptosporidiose zijn regelmatig gerapporteerd in Groot-Brittannie en de Verenigde Staten. De bron van de explosie kon soms achterhaald worden doordat Cryptosporidium oocysten in het zwembadwater of in het terugspoelwater van de zwembadfilters konden worden

  12. Cryptosporidium Infections Among Children in Peru

    Centers for Disease Control (CDC) Podcasts

    2008-09-25

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

  13. Sporulation dynamics of poultry Eimeria oocysts in Chennai.

    Science.gov (United States)

    Venkateswara Rao, P; Raman, M; Gomathinayagam, S

    2015-12-01

    The infective form of Eimeria is the highly resistant oocyst, which is shed in the faeces of infected animals. Present study was carried out to understand the sporulation dynamics of six Eimeria oocysts viz. E. acervulina, E. brunetti, E. maxima, E. mitis, E. necatrix and E. tenella in Chennai. Faecal samples of poultry were collected from various poultry farms located in and around Tamil Nadu. Oocysts of various Eimeria species were examined microscopically for sporulation on a 6 h interval basis till complete sporulation is acheived. The sporulation time recorded was 168, 120, 216, 192, 96 and 96 h for E. acervulina, E. brunetti, E. maxima, E. mitis, E. necatrix and E. tenella respectively. It can be concluded on comparison with previous studies that humid weather conditions delay the sporulation time and dry weather and wet litter is the ideal condition for rapid sporulation.

  14. Effects of Artemisia annua extracts on sporulation of Eimeria oocysts.

    Science.gov (United States)

    Fatemi, Ahmadreza; Razavi, Seyyed Mostafa; Asasi, Keramat; Goudarzi, Majid Torabi

    2015-03-01

    The present study aimed to compare the effect of different Artemisia annua extracts on sporulation rate of mixed oocysts of Eimeria acervulina, Eimeria necatrix, and Eimeria tenella. Three types of A. annua extracts including petroleum ether (PE), ethanol 96° (E), and water (W) extracts were prepared. Artemisinin, a sesquiterpene lactone endoperoxide derived from the A. annua analysis of each extract was done by high-performance liquid chromatography with ultraviolet detection (HPLC-UV). Fresh fecal samples containing three Eimeria species were floated and counted, and the oocysts were transferred into 50 tubes, each containing 10(5) oocysts per milliliter. Five tubes were control. Each of the other 45 tubes contained one of three doses (1 part per thousand (ppt), 2 ppt, and 5 ppt) and one of three extracts (PE, E, and W extracts) with five replications. The tubes were incubated for 48 h at 25-29 °C and aerated. Sporulation inhibition assay was used to evaluate the activity of extracts. The results showed that the E and PE extracts inhibit sporulation in 2 and 5 ppt concentrations, but the W extract stimulates it in all concentrations. The proportions of oocyst inhibition relative to control were 31 % (5 ppt) and 29 % (2 ppt) for PE and 34 % (5 ppt) and 46 % (2 ppt) for E extract. Furthermore, many oocysts in PE and E groups were wrinkled and contained abnormal sporocysts. The proportions of sporulation stimulation relative to control were 22 % (5 ppt), 24 % (2 ppt), and 27 % (1 ppt) in W extract. Our study is the first to demonstrate that all types of A. annua extracts do not necessarily have a similar activity, and the interaction of all contents and their relative concentrations is an important factor for sporulation stimulation or inhibition. It seems, some parts of unmetabolized excreted PE and E extracts could inhibit oocyst sporulation and eventually affect infection transmission.

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

    Directory of Open Access Journals (Sweden)

    Flavio Medeiros Paz e Silva

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

  16. Sporulation and survival of Toxoplasma gondii oocysts in different types of commercial cat litters

    Science.gov (United States)

    Toxoplasma gondii oocysts are environmentally resistant and can survive outdoors for months in the dry and cold climates. In the present study, sporulation and survival of T. gondii oocysts was studied in different types of cat litters commercially available in the US. Oocysts sporulated within 2-...

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

    Directory of Open Access Journals (Sweden)

    Traversa Donato

    2010-03-01

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

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

    Directory of Open Access Journals (Sweden)

    Roberta Dos Santos Toledo

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

  19. Detection and molecular diversity of Giardia duodenalis and Cryptosporidium spp. in sheltered dogs and cats in Northern Spain.

    Science.gov (United States)

    Gil, Horacio; Cano, Lourdes; de Lucio, Aida; Bailo, Begoña; de Mingo, Marta Hernández; Cardona, Guillermo A; Fernández-Basterra, José A; Aramburu-Aguirre, Juan; López-Molina, Nuria; Carmena, David

    2017-06-01

    Domestic dogs and cats may act as natural reservoirs of a large number of zoonotic pathogens, including the enteric parasites Giardia duodenalis and Cryptosporidium spp., the most relevant protozoan species causing gastrointestinal disease worldwide. A cross-sectional epidemiological study aiming to assess the prevalence and molecular diversity of G. duodenalis and Cryptosporidium spp. was conducted in an animal rescue centre in the province of Álava (Northern Spain). A total of 194 and 65 faecal dropping samples from individual dogs and cats, respectively, were collected between November 2013 and June 2016. G. duodenalis cysts and Cryptosporidium spp. oocysts were detected by direct fluorescence microscopy and PCR-based methods targeting the small subunit ribosomal RNA gene of these parasites. Overall, G. duodenalis and Cryptosporidium spp. were detected in 33% (63/194) and 4.1% (8/194) of dogs, and 9.2% (6/65) and 4.6% (3/65) of cats, respectively. G. duodenalis and Cryptosporidium co-infections were observed in 1.5% (3/194) of dogs, but not in cats. No significant differences in infection rates could be demonstrated among dogs or cats according to their sex, age group, status, or geographical origin. Multi-locus sequence-based genotyping of the glutamate dehydrogenase and β-giardin genes of G. duodenalis allowed the characterization of 19 canine isolates that were unambiguously assigned to sub-assemblages AII (n=7), BIII (n=1), and BIV (n=7), and assemblages C (n=3) and D (n=1). Two feline isolates were genotyped as assemblages A and F, respectively. No mixed assemblage or sub-assemblage infections were identified. C. canis (n=5) and C. hominis (n=1) were the Cryptosporidium species found in dogs, whereas C. felis (n=1) was identified in cats. The finding of G. duodenalis sub-assemblages AII, BIII, and BIV circulating in dogs (but not cats) may have zoonotic potential, although most of the AII and BIV isolates sub-genotyped corresponded to genetic variants not

  20. Macrophages facilitate the excystation and differentiation of Toxoplasma gondii sporozoites into tachyzoites following oocyst internalisation.

    Science.gov (United States)

    Freppel, Wesley; Puech, Pierre-Henri; Ferguson, David J P; Azas, Nadine; Dubey, Jitender P; Dumètre, Aurélien

    2016-09-19

    Toxoplasma gondii is a common parasite of humans and animals, which is transmitted via oocysts in cat faeces or tissue cysts in contaminated meat. The robust oocyst and sporocyst walls protect the infective sporozoites from deleterious external attacks including disinfectants. Upon oocyst acquisition, these walls lose their integrity to let the sporozoites excyst and invade host cells following a process that remains poorly understood. Given the resistance of the oocyst wall to digestive enzymes and the ability of oocysts to cause parenteral infections, the present study investigated the possible contribution of macrophages in supporting sporozoite excystation following oocyst internalisation. By using single cell micromanipulations, real-time and time-point imaging techniques, we demonstrated that RAW macrophages could interact rapidly with oocysts and engulfed them by remodelling of their actin cytoskeleton. Internalised oocysts were associated to macrophage acidic compartments and showed evidences of wall disruption. Sporozoites were observed in macrophages containing oocyst remnants or in new macrophages, giving rise to dividing tachyzoites. All together, these results highlight an unexpected role of phagocytic cells in processing T. gondii oocysts, in line with non-classical routes of infection, and open new perspectives to identify chemical factors that lead to oocyst wall disruption under physiological conditions.

  1. Comparison of Therapeutic Effect of Anti-Cryptosporidium Nano-Nitazoxanide (NTZ with Free form of this Drug in Neonatal Rat

    Directory of Open Access Journals (Sweden)

    F. Sedighi

    2016-07-01

    Full Text Available Introduction & Objective: Cryptosporidiosis caused by Cryptosporidium, which is a protozoan parasite, has a worldwide distribution. The infection is through fecal-oral route, direct or indi-rect contact, food or water. The treatment of cryptosporidiosis is difficult and the anti-parasitic agents are not effective. The purpose of this study was encapsulation of nitazoxanide in solid lipid nano-particles (SLN and investigation of its anti-Cryptosporidium effect and its comparison with free drug in the neonatal rat. Materials & Methods: Nitazoxanide was encapsulated by HPH method with 2 mg/Kg concentra-tion in SLN nanoparticles. The oocysts were collected from calves and purified by sucrose floatation. A total of 72 Wistar neonatal rats were categorized in 6 groups of 12 rats including four infected groups treated by free drug, encapsulated nano drug, colloidal carriers without drug (SLN and olive oil; an infected control group and a healthy control group that received PBS. 5 × 105 of oocyts inoculated orally into the sample groups. Finally, intestine of each rat was homogenized in PBS by rotor and the homogenized material was passed through a sieve. Then, floated oocysts in sucrose solution were counted by hemocytometer. Results: Treatment by nitazoxanide significantly decreased the number of parasites in the treatment groups. This decrease at day 6 was more than day 3. Nano nitazoxanide had more effects on parasites than free drug. This difference at day 3 of treatment was not significant (p= 0.182 but at day 6 was statistically significant (P< 0.001. Conclusion: Using nano-nitazoxanide could be a more effective way in the treatment of Cryp-tosporidium infections. (Sci J Hamadan Univ Med Sci 2016; 23 (2:134-140

  2. Point-of-Use Removal of Cryptosporidium parvum from Water: Independent Effects of Disinfection by Silver Nanoparticles and Silver Ions and by Physical Filtration in Ceramic Porous Media.

    Science.gov (United States)

    Abebe, Lydia S; Su, Yi-Hsuan; Guerrant, Richard L; Swami, Nathan S; Smith, James A

    2015-11-03

    Ceramic water filters (CWFs) impregnated with silver nanoparticles are a means of household-level water treatment. CWFs remove/deactivate microbial pathogens by employing two mechanisms: metallic disinfection and physical filtration. Herein we report on the independent effects of silver salt and nanoparticles on Cryptosporidium parvum and the removal of C. parvum by physical filtration in porous ceramic filter media. Using a murine (mouse) model, we observed that treatment of oocysts with silver nitrate and proteinate-capped silver nanoparticles resulted in decreased infection relative to untreated oocysts. Microscopy and excystation experiments were conducted to support the disinfection investigation. Heat and proteinate-capped silver-nanoparticle treatment of oocysts resulted in morphological modifications and decreased excystation rates of sporozoites. Subsequently, disk-shaped ceramic filters were produced to investigate the transport of C. parvum. Two factors were varied: sawdust size and clay-to-sawdust ratio. Five disks were prepared with combinations of 10, 16, and 20 mesh sawdust and sawdust percentage that ranged from 9 to 11%. C. parvum removal efficiencies ranged from 1.5 log (96.4%) to 2.1 log (99.2%). The 16-mesh/10% sawdust had the greatest mean reduction of 2.1-log (99.2%), though there was no statistically significant difference in removal efficiency. Based on our findings, physical filtration and silver nanoparticle disinfection likely contribute to treatment of C. parvum for silver impregnated ceramic water filters, although the contribution of physical filtration is likely greater than silver disinfection.

  3. DEVELOPMENT OF PATENT INFECTION IN IMMUNOSUPPRESSED C57BL/6 MICE WITH A SINGLE CRYPTOSPORIDIUM MELEAGRIDIS OOCYST. (R829180)

    Science.gov (United States)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  4. Seasonal prevalence and incidence of Cryptosporidium spp. and Giardia duodenalis and associated diarrhoea in children attending pre-schools in Kafue, Zambia

    DEFF Research Database (Denmark)

    Siwila, J.; Phiri, I.G.K.; Enemark, Heidi L.

    2011-01-01

    Prevalence, incidence and seasonal variation of Cryptosporidium and Giardia duodenalis were studied over a 12-month period in 100 children from four pre-schools in Kafue, Zambia. Questionnaire data and a single stool sample were collected monthly from each child. Samples were processed using...... a commercial kit (Meridian Diagnostics Inc., USA) and oo(cysts) visualised by immunofluorescence microscopy. Cryptosporidium was detected in 30.7% (241/786; 95% CI = 27.5-33.9) while G. duodenalis was detected in 29.0% (228/786; 95% CI = 25.8-32.2). A total of 86% experienced one or more episodes...... of cryptosporidiosis while 75% had giardiasis. Cumulative incidence per 100 children was 75.4 for Cryptosporidium and 49.0 for G. duodenalis. Both infections were significantly more common in the wet compared to the dry season (34.8%, 162/466 vs. 24.7%, 79/320, P = 0.003 and 35.2%, 164/466 vs. 20.0%, 64/320, P

  5. Evaluation and Comparison of Enzyme Immunoassay (Eia and Acid Fast Staining with Confirmation by Immunofluorescent Antibody Assay for Detection of Cryptosporidium Species in Infants and Young Children.

    Directory of Open Access Journals (Sweden)

    D Dorostcar Moghaddam

    2005-01-01

    Full Text Available Introduction: Cryptosporidiosis is prevalent world wide, causing a variety of problems ranging from acute, self-limiting diarrhea to fatal cases in immunocompromised persons, particulary those with acquired immunodeficiency (AIDS. Diagnosis of Cryptosporidium is made by identification of oocysts in stool specimens. The detection is most commonly made by the acid-fast staining method followed by microscopic examination which has low specificity and sensitivity. Material and Methods: In the present study, we evaluated diagnostic utility of a commercially available enzyme immunoassay (EIA, which detects Cryptosporidium-Specific antigen (CSA in 204 unprocessed stool specimens obtained from patients less than 3 years of age. Results: When compared with the routine screening procedure applied in this field study (screening by acid-fast staining and microscopy after concentration of positive results by IFA, both sensitivity and specificity were 98%. Of the 139 specimens negative by microscopy, 13 (9.3% were positive by EIA, 11 of which were confirmed by inhibition with antibody to Cryptosporidia-specific antigen. Conclusion: The EIA is an important tool for identifying Cryptosporidium in fecal specimens in field studies since it is sensitive, specific, simple to use and unaffected by the presence of a preservative.

  6. 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).

    Science.gov (United States)

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

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

    Czech Academy of Sciences Publication Activity Database

    Němejc, K.; Sak, Bohumil; Květoňová, Dana; Hanzal, V.; Janiszewski, P.; Forejtek, P.; Rajský, D.; Ravaszová, P.; McEvoy, J.; Kváč, Martin

    2013-01-01

    Roč. 197, 3-4 (2013), s. 504-508 ISSN 0304-4017 Grant - others:Jihočeská univerzita(CZ) 022/2010/Z; Jihočeská univerzita(CZ) 11/2013/Z Institutional support: RVO:60077344 Keywords : Central Europe * Cryptosporidium scrofarum * Cryptosporidium suis * Eurasian wild boar * PCR * SSU Subject RIV: EE - Microbiology, Virology Impact factor: 2.545, year: 2013

  8. Diarrhea due to Cryptosporidium parvum in immunocompromised ...

    African Journals Online (AJOL)

    Objective: The objective of this study is to search for Cryptosporidium parvum in Sudanese immunocompromised and immunocompetent patients presenting with diarrhea. Methods: Two hundred and thirteen stool specimens were collected from different groups of patients presenting with diarrhea and healthy control ...

  9. Emissie van Cryptosporidium en Giardia door landbouwhuisdieren

    NARCIS (Netherlands)

    Schrijven JF; Bruin HAM de; Engels GB; Leenen EJTM; MGB

    1999-01-01

    Het hier gepresenteerde deelonderzoek richt zich op de relatieve bijdrage van verschillende populaties landbouwhuisdieren via mest en afvalwater aan de totale emissie van Cryptosporidium en Giardia in Nederland. Vleeskalveren vormen per jaar in Nederland via hun mest een grote emissiebron van

  10. Cryptosporidium Pig Genotype II in Immunocompetent Man

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Květoňová, Dana; Sak, Bohumil; Ditrich, Oleg

    2009-01-01

    Roč. 15, č. 6 (2009), s. 982-983 ISSN 1080-6040 R&D Projects: GA ČR GP523/07/P117 Institutional research plan: CEZ:AV0Z60220518 Keywords : immunocompetent patients * cryptosporidiosis * Cryptosporidium pig genotype II Subject RIV: GJ - Animal Vermins ; Diseases , Veterinary Medicine Impact factor: 6.794, year: 2009

  11. Occurrence of Cryptosporidium andersoni in Brazilian cattle

    Science.gov (United States)

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

  12. Emergence of Cryptosporidium hominis Monkey Genotype II and Novel Subtype Family Ik in the Squirrel Monkey (Saimiri sciureus) in China.

    Science.gov (United States)

    Liu, Xuehan; Xie, Na; Li, Wei; Zhou, Ziyao; Zhong, Zhijun; Shen, Liuhong; Cao, Suizhong; Yu, Xingming; Hu, Yanchuan; Chen, Weigang; Peng, Gangneng

    2015-01-01

    A single Cryptosporidium isolate from a squirrel monkey with no clinical symptoms was obtained from a zoo in Ya'an city, China, and was genotyped by PCR amplification and DNA sequencing of the small-subunit ribosomal RNA (SSU rRNA), 70-kDa heat shock protein (HSP70), Cryptosporidium oocyst wall protein, and actin genes. This multilocus genetic characterization determined that the isolate was Cryptosporidium hominis, but carried 2, 10, and 6 nucleotide differences in the SSU rRNA, HSP70, and actin loci, respectively, which is comparable to the variations at these loci between C. hominis and the previously reported monkey genotype (2, 3, and 3 nucleotide differences). Phylogenetic studies, based on neighbor-joining and maximum likelihood methods, showed that the isolate identified in the current study had a distinctly discordant taxonomic status, distinct from known C. hominis and also from the monkey genotype, with respect to the three loci. Restriction fragment length polymorphisms of the SSU rRNA gene obtained from this study were similar to those of known C. hominis but clearly differentiated from the monkey genotype. Further subtyping was performed by sequence analysis of the gene encoding the 60-kDa glycoprotein (gp60). Maximum homology of only 88.3% to C. hominis subtype IdA10G4 was observed for the current isolate, and phylogenetic analysis demonstrated that this particular isolate belonged to a novel C. hominis subtype family, IkA7G4. This study is the first to report C. hominis infection in the squirrel monkey and, based on the observed genetic characteristics, confirms a new C. hominis genotype, monkey genotype II. Thus, these results provide novel insights into genotypic variation in C. hominis.

  13. MLST subtypes and population genetic structure of Cryptosporidium andersoni from dairy cattle and beef cattle in northeastern China's Heilongjiang Province.

    Directory of Open Access Journals (Sweden)

    Wei Zhao

    Full Text Available Cattle are the main reservoir host of C. andersoni, which shows a predominance in yearlings and adults of cattle. To understand the subtypes of C. andersoni and the population genetic structure in Heilongjiang Province, fecal specimens were collected from 420 dairy cattle and 405 beef cattle at the age of 12-14 months in eight cattle farms in five areas within this province and were screened for the presence of Cryptosporidium oocysts by microscopy after Sheather's sugar flotation technique. The average prevalence of Cryptosporidium spp. was 19.15% (158/825 and all the Cryptosporidium isolates were identified as C. andersoni by the SSU rRNA gene nested PCR-RFLP using SspI, VspI and MboII restriction enzymes. A total of 50 C. andersoni isolates were randomly selected and sequenced to confirm the RFLP results before they were subtyped by multilocus sequence typing (MLST at the four microsatellite/minisatellite loci (MS1, MS2, MS3 and MS16. Four, one, two and one haplotypes were obtained at the four loci, respectively. The MLST subtype A4,A4,A4,A1 showed an absolute predominance and a wide distribution among the six MLST subtypes obtained in the investigated areas. Linkage disequilibrium analysis showed the presence of a clonal population genetic structure of C. andersoni in cattle, suggesting the absence of recombination among lineages. The finding of a clonal population genetic structure indicated that the prevalence of C. andersoni in cattle in Heilongjiang Province is not attributed to the introduction of cattle. Thus, prevention and control strategies should be focused on making stricter measures to avoid the occurrence of cross-transmission and re-infection between cattle individuals. These molecular data will also be helpful to explore the source attribution of infection/contamination of C. andersoni and to elucidate its transmission dynamics in Heilongjiang Province, even in China.

  14. Global iTRAQ-based proteomic profiling of Toxoplasma gondii oocysts during sporulation.

    Science.gov (United States)

    Zhou, Chun-Xue; Zhu, Xing-Quan; Elsheikha, Hany M; He, Shuai; Li, Qian; Zhou, Dong-Hui; Suo, Xun

    2016-10-04

    Toxoplasma gondii is a medically and economically important protozoan parasite. However, the molecular mechanisms of its sporulation remain largely unknown. Here, we applied iTRAQ coupled with 2D LC-MS/MS proteomic analysis to investigate the proteomic expression profile of T. gondii oocysts during sporulation. Of the 2095 non-redundant proteins identified, 587 were identified as differentially expressed proteins (DEPs). Based on Gene Ontology enrichment and KEGG pathway analyses the majority of these DEPs were found related to the metabolism of amino acids, carbon and energy. Protein interaction network analysis generated by STRING identified ATP-citrate lyase (ACL), GMP synthase, IMP dehydrogenase (IMPDH), poly (ADP-ribose) glycohydrolase (PARG), and bifunctional dihydrofolate reductase-thymidylate synthase (DHFR-TS) as the top five hubs. We also identified 25 parasite virulence factors that were expressed at relatively high levels in sporulated oocysts compared to non-sporulated oocysts, which might contribute to the infectivity of mature oocysts. Considering the importance of oocysts in the dissemination of toxoplasmosis these findings may help in the search of protein targets with a key role in infectiousness and ecological success of oocysts, creating new opportunities for the development of better means for disease prevention. The development of new preventative interventions against T. gondii infection relies on an improved understanding of the proteome and chemical pathways of this parasite. To identify proteins required for the development of environmentally resistant and infective T. gondii oocysts, we compared the proteome of non-sporulated (immature) oocysts with the proteome of sporulated (mature, infective) oocysts. iTRAQ 2D-LC-MS/MS analysis revealed proteomic changes that distinguish non-sporulated from sporulated oocysts. Many of the differentially expressed proteins were involved in metabolic pathways and 25 virulence factors were identified

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

    Science.gov (United States)

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

    2016-04-01

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

  16. Study on the immunogenic ability of Eimeria tennela oocysts following treatment with gamma rays

    International Nuclear Information System (INIS)

    Penev, P.; Stefanova, M.

    1975-01-01

    Studied was the immunizing capacity of Eimeria tennela oocysts, treated with gamma rays at the rate of 6000 R, in 10- and 20-day-old chickens. The oocysts sporulated after treatment. Applied at the rate of 50000 R they showed lower virulence and were capable of inducing resistance to reinfection with non-irradiated oocysts at rates that were three times as much. Following reinfection some birds manifested subclinical coccidiosis but survived. This showed that the immunization with oocysts that had been irradiated with 6000 R had its peculiar aspect. (author)

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

    Science.gov (United States)

    Al-Mathal, Ebtisam M; Alsalem, Afaf M

    2012-07-01

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

  18. Cryptosporidiosis in broiler chickens in Zhejiang Province, China: molecular characterization of oocysts detected in fecal samples

    Directory of Open Access Journals (Sweden)

    Wang Lengmei

    2014-01-01

    Full Text Available Cryptosporidium is one of the most important parasites in poultry, and this pathogen can infect more than 30 avian species. The present study investigated the infection rate of Cryptosporidium among broiler chicken flocks. A total of 385 fecal samples from broiler chickens in 7 regions of Zhejiang Province collected from November 2010 to January 2012 were examined by microscopy. Thirty-eight (10% samples were positive for Cryptosporidium infection, and 3 genotypes (Cryptosporidium baileyi, Cryptosporidium meleagridis, and avian genotype II were identified by PCR and sequencing. A phylogenetic tree of the isolates was analyzed. These results suggest that cryptosporidiosis is widespread in poultry in Zhejiang Province, and is a potential threat to public health as well as the economy. This is the first report about the infection rate and molecular characterization of Cryptosporidium in broiler chickens in Zhejiang.

  19. Molecular epidemiology of Cryptosporidium spp. and Giardia spp. in mussels (Mytilus californianus) and California sea lions (Zalophus californianus) from Central California.

    Science.gov (United States)

    Adell, A D; Smith, W A; Shapiro, K; Melli, A; Conrad, P A

    2014-12-01

    Cryptosporidium and Giardia are of public health importance, with recognized transmission through recreational waters. Therefore, both can contaminate marine waters and shellfish, with potential to infect marine mammals in nearshore ecosystems. A 2-year study was conducted to evaluate the presence of Cryptosporidium and Giardia in mussels located at two distinct coastal areas in California, namely, (i) land runoff plume sites and (ii) locations near sea lion haul-out sites, as well as in feces of California sea lions (CSL) (Zalophus californianus) by the use of direct fluorescent antibody (DFA) detection methods and PCR with sequence analysis. In this study, 961 individual mussel hemolymph samples, 54 aliquots of pooled mussel tissue, and 303 CSL fecal samples were screened. Giardia duodenalis assemblages B and D were detected in hemolymph from mussels collected near two land runoff plume sites (Santa Rosa Creek and Carmel River), and assemblages C and D were detected in hemolymph from mussels collected near a sea lion haul-out site (White Rock). These results suggest that mussels are being contaminated by protozoa carried in terrestrial runoff and/or shed in the feces of CSL. Furthermore, low numbers of oocysts and cysts morphologically similar to Cryptosporidium and Giardia, respectively, were detected in CSL fecal samples, suggesting that CSL could be a source and a host of protozoan parasites in coastal environments. The results of this study showed that Cryptosporidium and Giardia spp. from the feces of terrestrial animals and CSL can contaminate mussels and coastal environments. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

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

    Science.gov (United States)

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

    2012-07-01

    Through the use of case-control analyses and quantitative microbial risk assessment (QMRA), relative risks of transmission of cryptosporidiosis have been evaluated (recreational water exposure vs. drinking water consumption) for a Canadian community with higher than national rates of cryptosporidiosis. A QMRA was developed to assess the risk of Cryptosporidium infection through the consumption of municipally treated drinking water. Simulations were based on site-specific surface water contamination levels and drinking water treatment log₁₀ reduction capacity for Cryptosporidium. Results suggested that the risk of Cryptosporidium infection via drinking water in the study community, assuming routine operation of the water treatment plant, was negligible (6 infections per 10¹³ persons per day--5th percentile: 2 infections per 10¹⁵ persons per day; 95th percentile: 3 infections per 10¹² persons per day). The risk is essentially nonexistent during optimized, routine treatment operations. The study community achieves between 7 and 9 log₁₀ Cryptosporidium oocyst reduction through routine water treatment processes. Although these results do not preclude the need for constant vigilance by both water treatment and public health professionals in this community, they suggest that the cause of higher rates of cryptosporidiosis are more likely due to recreational water contact, or perhaps direct animal contact. QMRA can be successfully applied at the community level to identify data gaps, rank relative public health risks, and forecast future risk scenarios. It is most useful when performed in a collaborative way with local stakeholders, from beginning to end of the risk analysis paradigm. © 2011 Society for Risk Analysis.

  1. Prevalence of Giardia and Cryptosporidium species in dog park attending dogs compared to non-dog park attending dogs in one region of Colorado.

    Science.gov (United States)

    Wang, Andrea; Ruch-Gallie, Rebecca; Scorza, Valeria; Lin, Philip; Lappin, Michael R

    2012-03-23

    Dog parks are very popular in urban areas, but there are no current studies attempting to correlate visits to dog parks and risk of colonization by enteric parasites. The purpose of this study was to determine whether dog park visitation is associated with an increased prevalence of enteric parasites or an increase in prevalence of gastrointestinal signs in dogs in northern Colorado. Feces from dogs owned by veterinary students or Veterinary Teaching Hospital staff members were submitted with a completed survey form detailing dog park attendance rates, fecal character scores, and other clinical information. Feces were examined microscopically for parasites after sugar centrifugation, for Giardia spp. cysts and Cryptosporidium spp. oocysts by a commercially available immunofluorescence assay (FA) and the FA positive samples were genotyped after PCR amplification. The Giardia assemblages were determined using the glutamate dehydrogenase (GDH) β-giardin and triose phosphate isomerase (TPI) genes and the Cryptosporidium species were determined using the heat shock protein-70 gene. A total of 129 fecal samples were assayed; 66 were from dog park attending dogs and 63 were from non-dog park-attending dogs. The overall parasite prevalence rate was 7.0% (9 of 129 samples). Dog park attending dogs were more likely to be positive for Giardia or Cryptosporidium than non-dog park-attending dogs (p=0.0279), but there was no association of gastrointestinal signs with dog park attendance or with fecal flotation or FA results. The five Giardia isolates were assemblage C and/or D and the one Cryptosporidium isolate was Ctenocephalides canis. Copyright © 2011 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2017-10-01

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

  3. Molecular characterization of Cryptosporidium parvum and Cryptosporidium hominis GP60 subtypes worldwide

    Directory of Open Access Journals (Sweden)

    Catalina Avendaño V

    2017-09-01

    Full Text Available Cryptosporidium is a zoonotic parasite very important in animal health as well as in public health. It is because this is one of the main causes of diarrhea in children, calves, lambs and other variety of youth mammalians in a lot of countries. The globalization has enabled the exchange of biological material in different regions worldwide, encouraging the spread of diseases and exposure to these biological agents to different environmental conditions, inducing adaptation through genetic changes. Based in the polymorphism of the gene for GP60, this review intended to present the distribution of Cryptosporidium parvum and Cryptosporidium hominis in humans and calves worldwide. The subtype that affects cattle more frequently corresponds to IIaA15G2R; while the subtype most frequently isolated from human samples is IaA19G2.

  4. Global Distribution, Public Health and Clinical Impact of the Protozoan Pathogen Cryptosporidium

    Directory of Open Access Journals (Sweden)

    Lorenza Putignani

    2010-01-01

    Full Text Available Cryptosporidium spp. are coccidians, oocysts-forming apicomplexan protozoa, which complete their life cycle both in humans and animals, through zoonotic and anthroponotic transmission, causing cryptosporidiosis. The global burden of this disease is still underascertained, due to a conundrum transmission modality, only partially unveiled, and on a plethora of detection systems still inadequate or only partially applied for worldwide surveillance. In children, cryptosporidiosis encumber is even less recorded and often misidentified due to physiological reasons such as early-age unpaired immunological response. Furthermore, malnutrition in underdeveloped countries or clinical underestimation of protozoan etiology in developed countries contribute to the underestimation of the worldwide burden. Principal key indicators of the parasite distribution were associated to environmental (e.g., geographic and temporal clusters, etc. and host determinants of the infection (e.g., age, immunological status, travels, community behaviours. The distribution was geographically mapped to provide an updated picture of the global parasite ecosystems. The present paper aims to provide, by a critical analysis of existing literature, a link between observational epidemiological records and new insights on public health, and diagnostic and clinical impact of cryptosporidiosis.

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

    Directory of Open Access Journals (Sweden)

    Teichmann Klaus

    2016-01-01

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

  6. Macrophages facilitate the excystation and differentiation of Toxoplasma gondii sporozoites into tachyzoites following oocyst internalization

    Science.gov (United States)

    Toxoplasma gondii is a common parasite of humans and domestic animals, which is transmitted via oocysts in cat faeces or tissue cysts in contaminated meat. The oocyst and sporocyst walls are multilayered polymeric structures that protect the infective sporozoites from deleterious physical and chemic...

  7. Cryptosporidium infections in children in Durban Seasonal variation ...

    African Journals Online (AJOL)

    One hundred and eleven of 1229 children (9%) aged < 10 years admitted to King Edward VIII Hospital, Durban, with gastro-enteritis over a period of 1 year were found to harbour Cryptosporidium. Of these, 96 (89,7%) were < 2 years of age. Cryptosporidium was the only potential pathogen identified in 80 of these patients ...

  8. Cryptosporidium infection in cattle in Ogun state, Nigeria | Akinkuotu ...

    African Journals Online (AJOL)

    The prevalence of Cryptosporidium spp. in cattle faeces in Ogun state, Nigeria was determined by a commercially produced enzyme-linked immunosorbent assay kit. Out of a total of 200 samples, 37.5% were positive for Cryptosporidium coproantigens. The highest rate of infection (78.1%) was observed in calves up to 3 ...

  9. Cryptosporidium infection in undernourished children with HIV/AIDS ...

    African Journals Online (AJOL)

    Background: AIDS and Protein energy malnutrition (PEM) severely impair the immune system Cryptosporidium has over the last two decades emerged as a life threatening disease. The study attempts to determine the prevalence of Cryptosporidium infection in malnourished children with HIV/AIDS. Method: Blood and stool ...

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

    Czech Academy of Sciences Publication Activity Database

    Holubová, Nikola; Sak, Bohumil; Horčičková, Michaela; Hlásková, Lenka; Květoňová, Dana; Menchaca, S.; McEvoy, J.; Kváč, Martin

    2016-01-01

    Roč. 115, č. 6 (2016), s. 2243-2251 ISSN 0932-0113 R&D Projects: GA ČR GA15-01090S Institutional support: RVO:60077344 Keywords : Cryptosporidium avium * morphology * molecular analyses * transmission studies * Cryptosporidium avian genotype V Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.329, year: 2016

  11. Cryptosporidium species and cattle: Implication for public health and ...

    African Journals Online (AJOL)

    Cryptosporidium species and cattle: Implication for public health and water - Short Communication. VA Pam, COE Onwuliri, DA Dakul, ICJ Omalu. Abstract. This paper presents a brief summary of the ecology of Cryptosporidium species in Calves and humans and the existing scientific evidence that addresses the claim that ...

  12. Cryptosporidium and Cryptosporidiosis in Calves at Jos, Northern ...

    African Journals Online (AJOL)

    Cryptosporidium and Cryptosporidiosis in Calves at Jos, Northern Nigeria. VA Pam, DA Dakul, COE Onwuliri. Abstract. This study investigated the occurrence of cryptosporidium and cryptosporidiosis in calves from Jos, Northern Nigeria. Two hundred fecal samples were collected from the calves, recruited for an all year ...

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

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Hofmannová, L.; Hlásková, Lenka; Květoňová, Dana; Vitovec, J.; McEvoy, J.; Sak, Bohumil

    2014-01-01

    Roč. 201, 1-2 (2014), s. 9-17 ISSN 0304-4017 R&D Projects: GA MŠk(CZ) LH11061 Institutional support: RVO:60077344 Keywords : Cryptosporidium erinacei * taxonomy * morphology * molecular analyses * transmission studies * Cryptosporidium hedgehog genotype Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.460, year: 2014

  14. Studies on the effect of gamma-rays irradiation on the virulence and immunogenicity of Eimeria tenella oocysts

    International Nuclear Information System (INIS)

    Hayat, Birjees

    1976-01-01

    Complete attenuation of the infective oocysts of Eimeria tanella was obtained with a gamma ray dose of 15000r. Above this dose, pathogenicity and the sensitivity of the disease decreased. There was no difference in the level of immunity induced with irradiated and non-irradiated oocysts, but the mortality with the irradiated oocysts was much lower. (ARA)

  15. Inactivation Data.xlsx

    Data.gov (United States)

    U.S. Environmental Protection Agency — The data set is a spreadsheet that contains results of inactivation experiments that were conducted to to determine the effectiveness of chlorine in inactivating B....

  16. Source water assessment and nonpoint sources of acutely toxic contaminants: A review of research related to survival and transport of Cryptosporidium parvum

    Science.gov (United States)

    Walker, Mark J.; Montemagno, Carlo D.; Jenkins, Michael B.

    1998-12-01

    Amendments to the Safe Drinking Water Act (PL-930123) in 1996 required that public water supply managers identify potential sources of contamination within contributing areas. Nonpoint sources of acutely toxic microbial contaminants, such as Cryptosporidium parvum, challenge current approaches to source identification and management as a first step toward developing management plans for public water supply protection. Little may be known about survival and transport in the field environment, prescribed practices may not be designed to manage such substances, and infective stages may be present in vast numbers and may resist water treatment and disinfection processes. This review summarizes research related to survival and transport of C. parvum oocysts, as an example of an acutely toxic contaminant with nonpoint sources in animal agriculture. It discusses ∥1) significance of infected domesticated animals as potential sources of C. parvum, (2) laboratory and field studies of survival and transport, and (3) approaches to source control in the context of public health protection.

  17. The first report on Cryptosporidium suis and Cryptosporidium pig genotype II in Eurasian wild boars (Sus scrofa) (Czech Republic)

    Czech Academy of Sciences Publication Activity Database

    Němejc, K.; Sak, Bohumil; Květoňová, Dana; Hanzal, V.; Jeníková, Martina; Kváč, Martin

    2012-01-01

    Roč. 184, 2/4 (2012), 122-125 ISSN 0304-4017 Grant - others:Mšk(CZ) 6007665806 Institutional research plan: CEZ:AV0Z60220518; CEZ:AV0Z50450515 Keywords : Cryptosporidium suis * Cryptosporidium pig genotype II * Eurasian wild boar * SSU * PCR Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.381, year: 2012

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

    Czech Academy of Sciences Publication Activity Database

    Chelladurai, J.J.; Clark, M.E.; Kváč, Martin; Holubová, Nikola; Khan, E.; Stenger, B.L.S.; Giddings, C.W.; McEvoy, J.

    2016-01-01

    Roč. 115, č. 5 (2016), s. 1901-1906 ISSN 0932-0113 Institutional support: RVO:60077344 Keywords : Cryptosporidium * Red-winged blackbird * Passerines * Cryptosporidium galli * Avian genotypeVI * Proventriculus * Intestine Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.329, year: 2016

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

    Science.gov (United States)

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

    2016-09-01

    Extensive genetic variation is observed within the genus Cryptosporidium and the distribution of Cryptosporidium species/genotypes in humans and animals appears to vary by geography and host species. To better understand the genetic diversity of Cryptosporidium spp. in horses and donkeys, we characterized five horse-derived and 82 donkey-derived Cryptosporidium isolates from five provinces or autonomous regions (Sichuan, Gansu, Henan, Inner Mongolia and Shandong) in China at the species/genotype and subtype levels. Three Cryptosporidium species/genotypes were identified based on the analysis of the SSU rRNA gene, including Cryptosporidium parvum (n=22), the Cryptosporidium horse genotype (n=4), and Cryptosporidium hominis (n=61). The identification of C. hominis was confirmed by sequence analysis of the HSP70 and actin genes. Subtyping using sequence analysis of the 60kDa glycoprotein gene identified 21 C. parvum isolates as subtype IIdA19G1, the four horse genotype isolates as subtypes VIaA15G4 (n=2) and VIaA11G3 (n=2), and the 61 C. hominis isolates as IkA16G1 (n=59) and IkA16 (n=2). The common finding of C. hominis reaffirms the heterogeneity of Cryptosporidium spp. in horses and donkeys and is possibly a reflection of endemic transmission of C. hominis in these animals. Data of the study suggest that horses and donkeys as companion animals may potentially transmit Cryptosporidium infections to humans. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Eimeria Oocyst Concentrations and Species Composition in Litter from Commercial Broiler Farms During Anticoccidial Drug or Live Eimeria Oocyst Vaccine Control Programs.

    Science.gov (United States)

    Jenkins, Mark C; Parker, Carolyn; Ritter, Donald

    2017-06-01

    The purpose of this study was to determine if Eimeria oocyst concentrations and species composition in commercial broiler house litter changed during different cycles of anticoccidial drug (ACD) or live Eimeria oocyst vaccine (VAC) control programs and if there was a correlation between Eimeria oocyst levels and broiler performance. Litter samples were collected from a total of 15 different broiler farms encompassing a total of 45 individual houses during at least one complete grow-out cycle over a 21-mo period. Of these 15 broiler farms, three were followed for the entire 21-mo period spanning three ACD and four VAC cycles. Samples were collected at 2, 4, and 7-8 wk of grow-out corresponding to starter, grower, and withdraw periods of the ACD cycle. On a number of occasions, litter samples were obtained just prior to chick placement. Eimeria oocysts were isolated from all samples, counted by microscopy, and extracted for DNA to identify Eimeria species by ITS1 PCR. In general, Eimeria oocyst concentration in litter reached peak levels at 2-4 wk of grow-out regardless of coccidiosis control measure being used. However, peak oocyst numbers were sometimes delayed until 7-8 wk, indicating some level of Eimeria spp. drug resistance or incomplete vaccine coverage. Eimeria maxima , Eimeria acervulina , Eimeria praecox, and Eimeria tenella were generally present in all samples, and no difference in the species composition was noted between houses on a particular farm. While Eimeria species composition was similar among houses, Eimeria spp. oocyst levels exhibited sporadic peaks in one house of a given location's houses. Of particular interest was the observed correlation between E. maxima oocyst abundance and chick mortality. However, no correlation was observed in E. maxima oocyst levels, and the performance parameters adjusted feed conversion ratio and average daily weight gain. This study showed that understanding the dynamics of Eimeria spp. oocyst levels and species

  1. Identification of Giardia lamblia and the human infectious-species of Cryptosporidium in drinking water resources in Western Saudi Arabia by nested-PCR assays.

    Science.gov (United States)

    Hawash, Y; Ghonaim, M; Hussein, Y; Alhazmi, A; Alturkistani, A

    2015-06-01

    The presence of Cryptosporidium and/or Giardia in drinking water represents a major public health problem. This study was the first report concerned with the occurrence of these protozoa in drinking water in Saudi Arabia. The study was undertaken in Al-Taif, a high altitude region, Western Saudi Arabia. Eight underground wells water, six desalinated water and five domestic brands of bottled water samples, 10 liter each, were monthly collected between May 2013 and April 2014. All samples (n = 228), were processed using an automated wash/elution station (IDEXX Laboratories, Inc.). Genomic DNA was directly isolated and purified from samples concentrates with QIAamp® Stool Mini Kit (Qiagen). The target protozoan DNA sequences were amplified using two previously published nested-PCR protocols. Of all the analyzed water, 31 samples (≈14%) were found contaminated with the target protozoa. Giardia lamblia was detected in ≈10% (7/72) of desalinated water and in ≈9% (9/96) of wells water. On the other hand, Cryptosporidium was identified in ≈8% (8/72) of desalinated water and in ≈7% (7/96) of wells water. All bottled water samples (n = 60) were (oo)cysts-free. Protozoan (oo)cysts were more frequently identified in water samples collected in the spring than in other seasons. The methodology established in our study proved sensitive, cost-effective and is amenable for future automation or semi-automation. For better understanding of the current situation that represent an important health threat to the local inhabitants, further studies concerned with (oo)cyst viability, infectivity, concentration and genotype identification are recommended.

  2. New view on the age-specificity of pig Cryptosporidium by species-specific primers for distinguishing Cryptosporidium suis and Cryptosporidium pig genotype II

    Czech Academy of Sciences Publication Activity Database

    Jeníková, M.; Němejc, K.; Sak, Bohumil; Květoňová, Dana; Kváč, Martin

    2011-01-01

    Roč. 176, 2/3 (2011), 120-125 ISSN 0304-4017 R&D Projects: GA ČR GP523/07/P117 Institutional research plan: CEZ:AV0Z60220518 Keywords : Cryptosporidium suis * Cryptosporidium pig genotype II * Mixed infection * Age-specificity * Species-specific primers Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.579, year: 2011

  3. Inactivation of Caliciviruses

    Directory of Open Access Journals (Sweden)

    Raymond Nims

    2013-03-01

    Full Text Available The Caliciviridae family of viruses contains clinically important human and animal pathogens, as well as vesivirus 2117, a known contaminant of biopharmaceutical manufacturing processes employing Chinese hamster cells. An extensive literature exists for inactivation of various animal caliciviruses, especially feline calicivirus and murine norovirus. The caliciviruses are susceptible to wet heat inactivation at temperatures in excess of 60 °C with contact times of 30 min or greater, to UV-C inactivation at fluence ≥30 mJ/cm2, to high pressure processing >200 MPa for >5 min at 4 °C, and to certain photodynamic inactivation approaches. The enteric caliciviruses (e.g.; noroviruses display resistance to inactivation by low pH, while the non-enteric species (e.g.; feline calicivirus are much more susceptible. The caliciviruses are inactivated by a variety of chemicals, including alcohols, oxidizing agents, aldehydes, and β-propiolactone. As with inactivation of viruses in general, inactivation of caliciviruses by the various approaches may be matrix-, temperature-, and/or contact time-dependent. The susceptibilities of the caliciviruses to the various physical and chemical inactivation approaches are generally similar to those displayed by other small, non-enveloped viruses, with the exception that the parvoviruses and circoviruses may require higher temperatures for inactivation, while these families appear to be more susceptible to UV-C inactivation than are the caliciviruses.

  4. Second outbreak of infection with a rare Cryptosporidium parvum genotype in schoolchildren associated with contact with lambs/goat kids at a holiday farm in Norway.

    Science.gov (United States)

    Lange, H; Johansen, O H; Vold, L; Robertson, L J; Anthonisen, I L; Nygard, K

    2014-10-01

    In March 2012, a second outbreak of Cryptosporidium parvum affected children following a stay at a holiday farm in Norway; the first outbreak occurred in 2009. We studied a cohort of 145 schoolchildren who had visited the farm, of which 40 (28%) were cases. Cryptosporidium oocysts were detected in faecal samples from humans, goat kids and lambs. Molecular studies revealed C. parvum subtype IIa A19G1R1 in all samples including human samples from the 2009 outbreak. A dose-response relationship was found between the number of optional sessions with animals and illness, increasing from two sessions [risk ratio (RR) 2·7, 95% confidence interval (CI) 0·6-11·5] to six sessions (RR 8·0, 95% CI 1·7-37·7). The occurrence of two outbreaks 3 years apart, with the same subtype of C. parvum, suggests that the parasite is established in the farm's environment. We recommend greater emphasis on hand hygiene and routines related to animal contact.

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

    Science.gov (United States)

    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.

  6. Occurrence of Giardia, Cryptosporidium, and Entamoeba in wild rhesus macaques (Macaca mulatta living in urban and semi-rural North-West India

    Directory of Open Access Journals (Sweden)

    John J. Debenham

    2017-04-01

    Full Text Available Giardia duodenalis, Cryptosporidium spp., and Entamoeba spp. are intestinal protozoa capable of infecting a range of host species, and are important causes of human morbidity and mortality. Understanding their epidemiology is important, both for public health and for the health of the animals they infect. This study investigated the occurrence of these protozoans in rhesus macaques (Macaca mulatta in India, with the aim of providing preliminary information on the potential for transmission of these pathogens between macaques and humans. Faecal samples (n = 170 were collected from rhesus macaques from four districts of North-West India. Samples were analysed for Giardia/Cryptosporidium using a commercially available direct immunofluorescent antibody test after purification via immunomagnetic separation. Positive samples were characterised by sequencing of PCR products. Occurrence of Entamoeba was investigated first by using a genus-specific PCR, and positive samples further investigated via species-specific PCRs for Entamoeba coli, Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii. Giardia cysts were found in 31% of macaque samples, with all isolates belonging to Assemblage B. Cryptosporidium oocysts were found in 1 sample, however this sample did not result in amplification by PCR. Entamoeba spp. were found in 79% of samples, 49% of which were positive for E. coli. Multiplex PCR for E. histolytica, E. dispar and E. moshkovskii, did not result in amplification in any of the samples. Thus in 51% of the samples positive at the genus specific PCR, the Entamoeba species was not identified. This study provides baseline information on the potential for transmission of these zoonotic parasites at the wildlife-human interface.

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

    Science.gov (United States)

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

    2015-07-01

    Members of the genus Cryptosporidium are waterborne protozoa of great health concern. Many studies have attempted to find appropriate surrogates for assessing Cryptosporidium filtration removal in porous media. In this study, we evaluated the filtration of Cryptosporidium parvum in granular limestone medium by the use of biotin- and glycoprotein-coated carboxylated polystyrene microspheres (CPMs) as surrogates. Column experiments were carried out with core material taken from a managed aquifer recharge site in Adelaide, Australia. For the experiments with injection of a single type of particle, we observed the total removal of the oocysts and glycoprotein-coated CPMs, a 4.6- to 6.3-log10 reduction of biotin-coated CPMs, and a 2.6-log10 reduction of unmodified CPMs. When two different types of particles were simultaneously injected, glycoprotein-coated CPMs showed a 5.3-log10 reduction, while the uncoated CPMs displayed a 3.7-log10 reduction, probably due to particle-particle interactions. Our results confirm that glycoprotein-coated CPMs are the most accurate surrogates for C. parvum; biotin-coated CPMs are slightly more conservative, while unmodified CPMs are markedly overly conservative for predicting C. parvum removal in granular limestone medium. The total removal of C. parvum observed in our study suggests that granular limestone medium is very effective for the filtration removal of C. parvum and could potentially be used for the pretreatment of drinking water and aquifer storage recovery of recycled water. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  8. Quantifying pathogen risks associated with potable reuse: A risk assessment case study for Cryptosporidium.

    Science.gov (United States)

    Amoueyan, Erfaneh; Ahmad, Sajjad; Eisenberg, Joseph N S; Pecson, Brian; Gerrity, Daniel

    2017-08-01

    This study evaluated the reliability and equivalency of three different potable reuse paradigms: (1) surface water augmentation via de facto reuse with conventional wastewater treatment; (2) surface water augmentation via planned indirect potable reuse (IPR) with ultrafiltration, pre-ozone, biological activated carbon (BAC), and post-ozone; and (3) direct potable reuse (DPR) with ultrafiltration, ozone, BAC, and UV disinfection. A quantitative microbial risk assessment (QMRA) was performed to (1) quantify the risk of infection from Cryptosporidium oocysts; (2) compare the risks associated with different potable reuse systems under optimal and sub-optimal conditions; and (3) identify critical model/operational parameters based on sensitivity analyses. The annual risks of infection associated with the de facto and planned IPR systems were generally consistent with those of conventional drinking water systems [mean of (9.4 ± 0.3) × 10 -5 to (4.5 ± 0.1) × 10 -4 ], while DPR was clearly superior [mean of (6.1 ± 67) × 10 -9 during sub-optimal operation]. Because the advanced treatment train in the planned IPR system was highly effective in reducing Cryptosporidium concentrations, the associated risks were generally dominated by the pathogen loading already present in the surface water. As a result, risks generally decreased with higher recycled water contributions (RWCs). Advanced treatment failures were generally inconsequential either due to the robustness of the advanced treatment train (i.e., DPR) or resiliency provided by the environmental buffer (i.e., planned IPR). Storage time in the environmental buffer was important for the de facto reuse system, and the model indicated a critical storage time of approximately 105 days. Storage times shorter than the critical value resulted in significant increases in risk. The conclusions from this study can be used to inform regulatory decision making and aid in the development of design or operational

  9. Toxoplasma gondii: A study of oocyst re-shedding in domestic cats.

    Science.gov (United States)

    Zulpo, Dauton Luiz; Sammi, Ana Sue; Dos Santos, Joeleni Rosa; Sasse, João Pedro; Martins, Thais Agostinho; Minutti, Ana Flávia; Cardim, Sérgio Tosi; de Barros, Luiz Daniel; Navarro, Italmar Teodorico; Garcia, João Luis

    2018-01-15

    The aim of the present study was to evaluate the re-shedding of T. gondii oocysts in cats fed tissue cysts of homologous and heterologous strains 12, 24 and 36 months after the first infection. Thirteen cats were used in the present study and were divided into four groups: G1 (n=2), G2 (n=3), G3 (n=5), and G4 (n=3). G1, G3 and G4 cats were infected with brain cysts of ME49 and G2 with TgDoveBr8, both genotype II strains of T. gondii. The G1 and G2 cats were re-infected after twelve months with brain cysts of VEG strain (genotype III), and G3 cats were re-infected with TgDoveBr1 (genotype II). The G3 cats were re-infected a third time after 24 months from the second infection, and the G4 cats were re-infected 36 months after the initial infection with cysts of the VEG strain. The cats' feces were evaluated using fecal flotation and genotyped with PCR-RFLP. The serological responses for IgM, IgA and IgG were determined by ELISA. All cats shed oocysts after the initial infection. Only one G1 cat shed oocysts when re-infected after twelve months with the VEG strain. No G2 cats excreted oocysts after the second infection with VEG. G3 cats, when re-infected after twelve months with the TgDoveBr1 strain, did not shed oocysts. However, when challenged after a third time with the VEG strain, three out of four cats shed oocysts. In the G4 group, when re-infected after thirty-six months with the VEG strain, two out of three cats shed oocysts. All oocyst samples were genotyped and characterized as the same genotype from the inoculum. Protection against oocyst re-excretion occurred in 90%, 25%, and 33.4% of cats after 12, 24, and 36 months from the initial infection, respectively. Therefore, the environmental contamination by oocysts from re-infected adult cats is only 30% lower than from kittens. In conclusion, the excretion of T. gondii oocysts was higher in experimentally re-infected cats throughout the years, especially when a heterologous strain was used. Copyright © 2017

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

    Science.gov (United States)

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

    2006-06-01

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

  11. Assessment of Cryptosporidium in wastewater reuse for drinking ...

    African Journals Online (AJOL)

    Assessment of Cryptosporidium in wastewater reuse for drinking water ... water supply needs and/or to reduce costs in many communities around the world. ... in a treatment plant geared for the production of drinking water from wastewater ...

  12. In vitro activity of natural and chemical products on sporulation of Eimeria species oocysts of chickens.

    Science.gov (United States)

    Gadelhaq, Sahar M; Arafa, Waleed M; Abolhadid, Shawky M

    2018-02-15

    This study was designed to investigate the ability of two herbal extracts and different chemical substances to inhibit or disrupt sporulation of Eimeria species oocysts of the chickens. The two herbal extracts were Allium sativum (garlic) and Moringa olifiera while the chemical substances included commercial disinfectants and diclazuril. Field isolates of Eimeria oocysts were propagated in chickens to obtain a continuous source of oocysts. The collected unsporulated oocysts (10 5 oocysts/5 ml) were dispensed into 5 cm Petri dish. Three replicates were used for each treatment. The treated oocysts were incubated for 48 h at 25-29 °C and 80% relative humidity. The results showed that herbal extracts, the commercial recommended dose of Dettol, TH4, Phenol, Virkon ® S, and Diclazuril 20% have no effect on the sporulation. While Sodium hypochlorite showed a significant degree of sporulation inhibition reached to 49.67%. Moreover, 70% ethanol, and 10% formalin showed 100% sporulation inhibition. It was concluded that 70% ethanol and 10% formalin are the most effective methods to inhibit Eimeria species sporulation. Copyright © 2017 Elsevier B.V. All rights reserved.

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

    OpenAIRE

    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 (Lampropel...

  14. Cryptosporidium Source Tracking in the Potomac River Watershed▿

    OpenAIRE

    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...

  15. Hypothesis: Cryptosporidium genetic diversity mirrors national disease notification rate

    OpenAIRE

    Takumi, Katsuhisa; Cacci?, Simone M.; van der Giessen, Joke; Xiao, Lihua; Sprong, Hein

    2015-01-01

    Background Cryptosporidiosis is a gastrointestinal disease affecting many people worldwide. Disease incidence is often unknown and surveillance of human cryptosporidiosis is installed in only a handful of developed countries. A genetic marker that mirrors disease incidence is potentially a powerful tool for monitoring the two primary human infected species of Cryptosporidium. Methods We used the molecular epidemiological database with Cryptosporidium isolates from ZoopNet, which currently con...

  16. Species-specific escape of Plasmodium sporozoites from oocysts of avian, rodent, and human malarial parasites.

    Science.gov (United States)

    Orfano, Alessandra S; Nacif-Pimenta, Rafael; Duarte, Ana P M; Villegas, Luis M; Rodrigues, Nilton B; Pinto, Luciana C; Campos, Keillen M M; Pinilla, Yudi T; Chaves, Bárbara; Barbosa Guerra, Maria G V; Monteiro, Wuelton M; Smith, Ryan C; Molina-Cruz, Alvaro; Lacerda, Marcus V G; Secundino, Nágila F C; Jacobs-Lorena, Marcelo; Barillas-Mury, Carolina; Pimenta, Paulo F P

    2016-08-02

    Malaria is transmitted when an infected mosquito delivers Plasmodium sporozoites into a vertebrate host. There are many species of Plasmodium and, in general, the infection is host-specific. For example, Plasmodium gallinaceum is an avian parasite, while Plasmodium berghei infects mice. These two parasites have been extensively used as experimental models of malaria transmission. Plasmodium falciparum and Plasmodium vivax are the most important agents of human malaria, a life-threatening disease of global importance. To complete their life cycle, Plasmodium parasites must traverse the mosquito midgut and form an oocyst that will divide continuously. Mature oocysts release thousands of sporozoites into the mosquito haemolymph that must reach the salivary gland to infect a new vertebrate host. The current understanding of the biology of oocyst formation and sporozoite release is mostly based on experimental infections with P. berghei, and the conclusions are generalized to other Plasmodium species that infect humans without further morphological analyses. Here, it is described the microanatomy of sporozoite escape from oocysts of four Plasmodium species: the two laboratory models, P. gallinaceum and P. berghei, and the two main species that cause malaria in humans, P. vivax and P. falciparum. It was found that sporozoites have species-specific mechanisms of escape from the oocyst. The two model species of Plasmodium had a common mechanism, in which the oocyst wall breaks down before sporozoites emerge. In contrast, P. vivax and P. falciparum sporozoites show a dynamic escape mechanism from the oocyst via polarized propulsion. This study demonstrated that Plasmodium species do not share a common mechanism of sporozoite escape, as previously thought, but show complex and species-specific mechanisms. In addition, the knowledge of this phenomenon in human Plasmodium can facilitate transmission-blocking studies and not those ones only based on the murine and avian models.

  17. Immunizing potential of sporulated oocysts of Eimeria nieschulzi exposed to heat and 60Co gamma-radiation

    International Nuclear Information System (INIS)

    Conder, G.A.; Duszynski, D.W.

    1977-01-01

    Sporulated oocysts of Eimeria nieschulzi Dieben 1924, a rat coccidium, were exposed to radiation, heat, or both in an effort to attenuate the parasite. Moderate levels of each treatment or combination thereof attenuated the parasite, reduced pathogenesis (as judged by oocyst discharge during primary infection), and produced immunity to challenge when the oocysts were subsequently inoculated into rats. Thus, heat- and/or radiation-treated E. nieschulzi oocysts fed to rats could reduce pathogenesis during a primary infection and yet give good homologous protection

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

    Science.gov (United States)

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

    2017-06-01

    The role of pet dogs and cats as suitable source of human infections by the diarrheagenic protozoan parasites Giardia duodenalis and Cryptosporidium spp. has been a topic of intense debate for long time and still remains a largely unsolved problem. In this cross-sectional molecular epidemiological survey we attempted to investigate whether zoonotic (or zooanthroponotic) disease transmission was occurring among humans and domestic dogs and cats sharing the same spatial and temporal setting in both rural and urban areas of the province of Álava, Northern Spain. A total of 268 (including 179 human, 55 canine, and 34 feline) individual faecal specimens were obtained from 63 family households during February-March and November-December 2014. Detection of G. duodenalis cysts and Cryptosporidium spp. oocysts was achieved by direct fluorescence microscopy (DFAT) and PCR-based methods targeting the small subunit (SSU) ribosomal RNA gene of the parasites. Giardia-positive isolates were subsequently sub-genotyped at the glutamate dehydrogenase (GDH) and β-giardin (BG) genes. Overall, G. duodenalis infections were identified in 3.4% (6/179) of humans, 29% (16/55) of dogs, and 5.9% (2/34) of cats, respectively. Cryptosporidium spp. infections were detected in 1.1% (2/179) of humans, 5.5% (3/55) of dogs, and 8.8% (3/34) of cats, respectively. Simultaneous infections in human and canine/feline hosts by G. duodenalis or Cryptosporidium spp. were only demonstrated in a single household in which a cat and its owner tested positive for Cryptosporidium by DFAT, but this result could not be confirmed by SSU-PCR. Infections were homogeneously distributed among the studied human or animal populations irrespectively of their sex, age group, or geographical region of origin. Inadequate washing of raw vegetables and fruits was the only risk factor significantly associated to a higher likelihood of having human giardiosis/cryptosporidiosis. Molecular characterization of G. duodenalis

  19. Comparison between two commercially available serological tests and polymerase chain reaction in the diagnosis of Cryptosporidium in animals and diarrhoeic children.

    Science.gov (United States)

    Helmy, Yosra A; Krücken, Jürgen; Nöckler, Karsten; von Samson-Himmelstjerna, Georg; Zessin, Karl-H

    2014-01-01

    For the detection of Cryptosporidium species in 804 animals and 165 diarrhoeic children (tests, the RIDASCREEN® Cryptosporidium test [enzyme immunoassay (EIA)] and the RIDA®QUICK Cryptosporidium/Giardia Combi [immuno-chromatographic test (ICT)] as well as polymerase chain reaction (PCR) were used. Prevalence of Cryptosporidium was 15.0, 19.5 and 32.3% in animals and 2.4, 6.7 and 49.1% in children using EIA, ICT and PCR, respectively.Using PCR as reference method, animal samples sensitivity (Se) of the EIA was 46.5% when questionable samples were considered positive, whereas specificity (Sp) was 100%. Se of the ICT was 60.4% while Sp was 100%. Positive predictive values (PPVs) for both EIA and ICT test were 100%, and negative predictive values (NPVs) for EIA were 79.7 and 84.1% for ICT. For the children samples, the Se of EIA was 5%, Sp was 100%, PPV was 100% and NPV was 52.2%, while the Se of ICT was 13.6%, Sp was 100%, PPV was 100% and NPV was 54.6%.The Kappa score of agreement between PCR and ICT was 67.4%, 54.1% between PCR and EIA and 84.4% between ICT and EIA. Until the second serial dilution of the EIA and ICT test, 9 × 10(3) oocysts/μl of Cryptosporidia was detected, whereas in PCR, they were detected until the sixth serial dilution. Copro-antigen tests were easy to perform and less time-consuming but less sensitive compared to PCR. They obviously are best applicable for screening and epidemiological studies of large numbers of subjects, for batch specimen processing and in isolated or rural areas where reliable tests like PCR are unfeasible. When in children, a single stool sample is used for the diagnosis of clinical cases; better results can be obtained when non-standardized PCR due low specificity is coupled with copro-antigen tests.

  20. Development of Eimeria nieschulzi (Coccidia, Apicomplexa Gamonts and Oocysts in Primary Fetal Rat Cells

    Directory of Open Access Journals (Sweden)

    Hong Chen

    2013-01-01

    Full Text Available The in vitro production of gametocytes and oocysts of the apicomplexan parasite genus Eimeria is still a challenge in coccidiosis research. Until today, an in vitro development of gametocytes or oocysts had only been shown in some Eimeria species. For several mammalian Eimeria species, partial developments could be achieved in different cell types, but a development up to gametocytes or oocysts is still lacking. This study compares several permanent cell lines with primary fetal cells of the black rat (Rattus norvegicus concerning the qualitative in vitro development of the rat parasite Eimeria nieschulzi. With the help of transgenic parasites, the developmental progress was documented. The selected Eimeria nieschulzi strain constitutively expresses the yellow fluorescent protein and a macrogamont specific upregulated red tandem dimer tomato. In the majority of all investigated host cells the development stopped at the second merozoite stage. In a mixed culture of cells derived from inner fetal organs the development of schizont generations I-IV, macrogamonts, and oocysts were observed in crypt-like organoid structures. Microgamonts and microgametes could not be observed and oocysts did not sporulate under air supply. By immunohistology, we could confirm that wild-type E. nieschulzi stages can be found in the crypts of the small intestine. The results of this study may be helpful for characterization of native host cells and for development of an in vitro cultivation system for Eimeria species.

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

    Science.gov (United States)

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

    2017-01-01

    The present review discusses the findings of cryptosporidiosis research conducted in cattle in China and highlights the currently available information on Cryptosporidium epidemiology, genetic diversity, and distribution in China, which is critical to understanding the economic and public health importance of cryptosporidiosis transmission in cattle. To date, 10 Cryptosporidium species have been detected in cattle in China, with an overall infection rate of 11.9%. The highest rate of infection (19.5%) was observed in preweaned calves, followed by that in juveniles (10.69%), postweaned juveniles (9.0%), and adult cattle (4.94%). The dominant species were C. parvum in preweaned calves and C. andersoni in postweaned, juvenile, and adult cattle. Zoonotic Cryptosporidium species (C. parvum and C. hominis) were found in cattle, indicating the possibility of transmission between humans and cattle. Different cattle breeds had significant differences in the prevalence rate and species of Cryptosporidium. This review demonstrates an age-associated, breed-associated, and geographic-related occurrence of Cryptosporidium and provides references for further understanding of the epidemiological characteristics, and for preventing and controlling the disease. © C. Gong et al., published by EDP Sciences, 2017.

  2. Interactions between Cryptosporidium parvum and the Intestinal Ecosystem

    KAUST Repository

    Douvropoulou, Olga

    2017-04-01

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

  3. Genetic diversity of Cryptosporidium spp. including novel identification of the Cryptosporidium muris and Cryptosporidium tyzzeri in horses in the Czech Republic and Poland

    Czech Academy of Sciences Publication Activity Database

    Wagnerová, Pavla; Sak, Bohumil; McEvoy, J.; Rost, M.; Perec Matysiak, A.; Ježková, J.; Kváč, Martin

    2015-01-01

    Roč. 114, č. 4 (2015), s. 1619-1624 ISSN 0932-0113 R&D Projects: GA ČR GA15-01090S Institutional support: RVO:60077344 Keywords : horse * Cryptosporidium * SSU * gp60 * MLST Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.027, year: 2015

  4. Presencia de Giardia lamblia y Cryptosporidium spp. en aguas residuales depuradas reutilizadas para riego agrícola en la isla de Tenerife, España. Efectos del transporte a larga distancia sobre la calidad del agua reutilizada Giardia lamblia and Cryptosporidium spp. Presence in treated wastewater reutilised for irrigation in Tenerife island, Spain. Long-distance transport effects in the reutilised water quality

    Directory of Open Access Journals (Sweden)

    Escolástico Aguiar González

    2002-06-01

    habitats. These protozoan investigation and detection have acquired importance in the last years due to their dispersion forms, which show resistance to the habitual treatments of potabilization and purification, and their classification as emerging pathogens which are the causative agents of important hydrical transmission outbreaks.In our work, Giardia lamblia cyst and Cryptosporidium spp. oocyst presence in Santa Cruz de Tenerife treated wastewater which is transported to the South of the island to be reutilised in agriculture. Furthermore, transport, storage and advanced treatment effects against cyst and oocyst concentration throughout the system and relation existence with other bacteriological and physical-chemical parameters, are also investigated.The obtained results demonstrate variable behaviours in cyst and oocyst concentration against applied treatments, and the depurative effect of the long-distance transport which seems to have in treated wastewater.There was not found any relation between cyst and oocyst concentration in treated wastewater and the traditional indicators of faecal contamination.

  5. Absorption and deposition of xanthophylls in broilers challenged with three dosages of Eimeria acervulina oocysts.

    Science.gov (United States)

    Hernández-Velasco, X; Chapman, H D; Owens, C M; Kuttappan, V A; Fuente-Martínez, B; Menconi, A; Latorre, J D; Kallapura, G; Bielke, L R; Rathinam, T; Hargis, B M; Tellez, G

    2014-01-01

    1. An experiment was designed to evaluate the effect of different doses of oocysts of Eimeria acervulina on intestinal absorption and skin deposition of xanthophylls (XAs) in broilers. 2. A total of 192 broiler chickens were randomly assigned to 4 groups: an uninfected control group and three groups inoculated with either 1 × 10(2), 1 × 10(4) or 1 × 10(5) sporulated oocysts of E. acervulina by gavaging at 21 d. There were 4 replicate pens (2 male and 2 female) per group. 3. Plasma xanthophyll (PX) and skin yellowness (SY) were measured in live birds weekly. At 42 d of age, SY was measured in the breast and abdomen after chilling and in the breast 24 h post-processing on refrigerated carcasses. 4. In general, in all challenged treatments, and for the duration of the study, the average PX decreased by 0.02 μg/ml (R(2) = 61.6%) for every 1000 inoculated oocysts, whereas PX increased by 1.26 μg/ml/d in uninfected birds. 5. The average SY in live birds from 21 to 42 d of age decreased by 0.019 b*/every 1000 oocysts administered, while SY of uninfected controls increased by 0.57 b*/d. It was also noted that in all treatments females had a greater SY (6.17 b*) than males for the duration of the study. The SY of the breast and abdomen was correlated (r = 0.76) in chilled carcasses. Breast SY in 24 h refrigerated carcasses was greater in the control group and for female birds. 6. Oocyst excretion was different between inoculated treatments only on 7 d post-inoculation (PI). Coccidia lesion scores in the duodenum averaged 1+ in infected birds and 2+ in birds given the highest oocyst dose.

  6. Immunoproteomic analysis of proteins from unsporulated Oocysts of Eimeria tenella in MALDI TOF/TOF tandem mass spectrometry

    Science.gov (United States)

    Immunoproteomic approaches were conducted to identify antigenic proteins from the total proteins of unsporulated oocysts of Eimeria tenella (E. tenella). Approximately 101 protein spots were recognized by chicken sera infected experimentally with E. tenella. Fourty-six spots of unsporulated oocysts ...

  7. The kinetics of oocyst shedding and sporulation in two immunologically distinct strains of Eimeria maxima, GS and M6.

    Science.gov (United States)

    Al-Badri, Riadh; Barta, John Robert

    2012-11-01

    The kinetics of oocyst shedding and sporulation of two immunologically distinct strains of Eimeria maxima (GS and M6) were compared. Both strains had a prepatent period of approximately 120 h followed by peak oocyst shedding at 144-150 h post inoculation. Mean total oocyst output determined for each strain demonstrated that the fecundity of the M6 strain (12.8 × 10(3) ± 1.95) of E. maxima was roughly twice that of the GS strain (6.9 × 10(3) ± 3.33) when inoculated at the rate of 1,000 infective oocysts per bird. The process of oocyst sporulation was followed by repetitive sampling of sporulating oocysts at 26 °C with aeration over a 138 hour period. Sporulation was divided into five morphologically distinguishable stages whose abundance peaked at the following times during sporulation: unsporulated oocysts at 0 h; sporoblast anlagen at 18 h; sporoblasts without sporocyst walls at 22 h; and sporocysts without mature sporozoites at 38 h. The time to 50 % sporulation of E. maxima oocysts observed in the present study was approximately 53 h for both strains and all viable oocysts had completed sporulation by 60 h. In the present study, the prepatent periods, duration of oocyst shedding, and the relative kinetics of sporulation of the GS and M6 strains of E. maxima were found to be virtually identical despite the immunological distinctiveness of these two parasite strains.

  8. Detection of Cryptosporidium sp. in non diarrheal faeces from children, in a day care center in the city of São Paulo, Brazil Detecção de Cryptosporidium sp. em fezes não diarréicas de crianças, em uma escola de educação infantil de São Paulo, Brasil

    Directory of Open Access Journals (Sweden)

    Therezinha T. Carvalho-Almeida

    2006-02-01

    Full Text Available The protozoan Cryptosporidium sp. has been frequently detected in faeces from children with persistent diarrhoea. This work achieved to investigate an outbreak of cryptosporidiosis, in a day care center, attending children of high socio-economic level, between 0 and six years old. The outbreak was detected through the network of public health, when stool samples, not diarrhoeic, were examined at the Parasitology Service of the Adolfo Lutz Institute. Among the 64 examined children, 13 (20.3% showed oocysts of Cryptosporidium sp. in the faeces examined by Kinyoun technique: seven children one year old, three, two years old and three, three years old. Among the 23 examined adults, only a 22 years old woman, possibly having an immunocomprometiment, was positive. Clinical and epidemiological aspects were investigated by questionnaires, highlighting the occurrence of the outbreak in a very dry period.O protozoário Cryptosporidium sp. tem sido detectado com freqüência em fezes de crianças com diarréia persistente. Este trabalho teve como objetivo investigar um surto de criptosporidiose, em uma escola de educação infantil, que atende crianças de classe média alta, de 0 a seis anos de idade. O surto foi detectado a partir de amostras fecais não diarréicas encaminhadas ao Instituto Adolfo Lutz através da rede de saúde pública. Das 64 crianças 13 (20,3% apresentaram oocistos de Cryptosporidium nas fezes, examinadas pelo método de Kinyoun, sendo sete crianças com um ano de idade, três com dois anos de idade e três, com três anos de idade. Dos 23 adultos examinados, apenas uma mulher de 22 anos, com possível imunocomprometimento, foi positiva. Aspectos epidemiológicos e clínicos foram investigados por meio de questionários, ressaltando-se que o surto ocorreu em período muito seco.

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

    DEFF Research Database (Denmark)

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

    2003-01-01

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

  10. Ultraviolet inactivation of papain

    International Nuclear Information System (INIS)

    Baugher, J.F.; Grossweiner, L.I.

    1975-01-01

    Flash photolysis transient spectra (lambda > 250 nm) of aqueous papain showed that the initial products are the neutral tryptophan radical Trp (lambdasub(max) 510 nm), the tryptophan triplet state 3 Trp (lambdasub(max) 460 nm), the disulfide bridge electron adduct -SS - - (lambdasub(max) 420 nm) and the hydrated electron esub(aq) - . The -SS - - yield was not altered by nitrous oxide or air, indicating that the formation of this product does not involve electrons in the external medium. The original papain preparation was activated by irradiating under nitrogen. The action spectrum supports previous work attributing the low initial activity to blocking of cysteinyl site 25 with a mixed disulfide. Flask lamp irradiation in nitrogen led to activation at low starting activities and inactivation at higher starting activities, while only inactivation at the same quantum yield was observed with air saturation. The results are consistent with photoionization of an essential tryptophyl residue as the key inactivating step. (author)

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

    Science.gov (United States)

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

    2005-01-01

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

  12. Molecular analysis of single oocyst of Eimeria by whole genome amplification (WGA) based nested PCR.

    Science.gov (United States)

    Wang, Yunzhou; Tao, Geru; Cui, Yujuan; Lv, Qiyao; Xie, Li; Li, Yuan; Suo, Xun; Qin, Yinghe; Xiao, Lihua; Liu, Xianyong

    2014-09-01

    PCR-based molecular tools are widely used for the identification and characterization of protozoa. Here we report the molecular analysis of Eimeria species using combined methods of whole genome amplification (WGA) and nested PCR. Single oocyst of Eimeria stiedai or Eimeriamedia was directly used for random amplification of the genomic DNA with either primer extension preamplification (PEP) or multiple displacement amplification (MDA), and then the WGA product was used as template in nested PCR with species-specific primers for ITS-1, 18S rDNA and 23S rDNA of E. stiedai and E. media. WGA-based PCR was successful for the amplification of these genes from single oocyst. For the species identification of single oocyst isolated from mixed E. stiedai or E. media, the results from WGA-based PCR were exactly in accordance with those from morphological identification, suggesting the availability of this method in molecular analysis of eimerian parasites at the single oocyst level. WGA-based PCR method can also be applied for the identification and genetic characterization of other protists. Copyright © 2014 Elsevier Inc. All rights reserved.

  13. Quantitative characterization, classification and reconstruction of oocyst shapes of Eimeria species from cattle

    DEFF Research Database (Denmark)

    Sommer, C.

    1998-01-01

    by multivariate statistical techniques. The morphology of 810 Eimeria specimens was defined in binary (b/w) digital images by pixels of their oocyst outline. A Fourier transform of pixel positions yielded size and shape features. To classify coccidia, the quantitative data were employed in an agglomerative...

  14. Crystal structure of Cryptosporidium parvum pyruvate kinase.

    Directory of Open Access Journals (Sweden)

    William J Cook

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

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

    Science.gov (United States)

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

    2013-09-01

    We report for the first time on the comparative use of pulsed-plasma gas-discharge (PPGD) and pulsed UV light (PUV) for the novel destruction of the waterborne enteroparasite Cryptosporidium parvum. It also describes the first cyto-, geno- and ecotoxicological assays undertaken to assess the safety of water decontaminated using PPGD and PUV. During PPGD treatments, the application of high voltage pulses (16 kV, 10 pps) to gas-injected water (N2 or O2, flow rate 2.5L/min) resulted in the formation of a plasma that generated free radicals, ultraviolet light, acoustic shock waves and electric fields that killed ca. 4 log C. parvum oocysts in 32 min exposure. Findings showed that PPGD-treated water produced significant cytotoxic properties (as determined by MTT and neutral red assays), genotoxic properties (as determined by comet and Ames assays), and ecotoxic properties (as determined by Microtox™, Thamnotox™ and Daphnotox™ assays) that are representative of different trophic levels in aquatic environment (pozone (0.8 mg/L) and/or dissociated nitric and nitrous acid that contributed to the observed disinfection and toxicity. Chemical analysis of PPGD-treated water revealed increasing levels of electrode metals that were present at ≤ 30 times the tolerated respective values for EU drinking water. PUV-treated water did not exhibit any toxicity and was shown to be far superior to that of PPGD for killing C. parvum oocysts taking only 90 s of pulsing [UV dose of 6.29 μJ/cm(2)] to produce a 4-log reduction compared to a similar reduction level achieved after 32min PPGD treatment as determined by combined in vitro CaCo-2 cell culture-qPCR. © 2013. Published by Elsevier B.V. All rights reserved.

  16. Occurrence and molecular characterization of Cryptosporidium sp. in sheep

    Directory of Open Access Journals (Sweden)

    Alessandra Snak

    2017-08-01

    Full Text Available Considered a zoonosis of utmost importance, cryptosporidiosis has a worldwide distribution and can infect mammals, birds, reptiles, and amphibians. It is caused by a highly resistant protozoan present in the environment and can cause death in immunosuppressed individuals and pups, as well as in farm animals such as cattle and sheep, generating losses. The aim of this study was to investigate the presence of Cryptosporidium spp. in sheep feces from the farms of Western Paraná, which have different management styles, and compare the results with their respective management methods. One hundred and forty-four stool samples were collected (69 from Property 1 and 75 from Property 2 and analyzed using a fecal smear on slides after staining by the modified Ziehl-Neelsen method. Samples tested positive by this method were subjected to nested PCR and the products obtained were sent for sequencing to determine the species. While 82.60% of the samples from Property 1 were tested positive, only 36% of the samples from Property 2 were tested positive. On analyzing the sequencing data, it was observed that the Cryptosporidium species of samples from Property 1 showed high similarity to Cryptosporidium xiaoi and those from Property 2, to Cryptosporidium ubiquitum. The reason for divergence in results can be attributed to differences in management systems adopted by each property, thus showing the importance of detecting carrier animals, as they can contaminate the environment, especially the water sources, and spread the disease to humans and other animals.

  17. Genetic diversity of Cryptosporidium spp. in captive reptiles

    Czech Academy of Sciences Publication Activity Database

    Xiao, L.; Ryan, U. M.; Graczyk, T. K.; Limor, J.; Li, L.; Kombert, M.; Junge, R.; Sulaiman, I. M.; Zhou, L.; Arrowood, M. J.; Koudela, Břetislav; Modrý, David; Lal, A. A.

    2004-01-01

    Roč. 70, č. 2 (2004), s. 891-899 ISSN 0099-2240 R&D Projects: GA ČR GA524/00/P015 Institutional research plan: CEZ:AV0Z6022909 Keywords : Cryptosporidium * reptiles * genetic diversity Subject RIV: EG - Zoology Impact factor: 3.810, year: 2004

  18. Cryptosporidium from tortoises: Genetic characterisation, phylogeny and zoonotic implications

    Czech Academy of Sciences Publication Activity Database

    Traversa, D.; Iorio, R.; Otranto, D.; Modrý, David; Šlapeta, J.

    2008-01-01

    Roč. 22, č. 2 (2008), s. 122-128 ISSN 0890-8508 Institutional research plan: CEZ:AV0Z60220518 Keywords : Cryptosporidium * tortoises * COWP * Testudo * epidemiology * 18S SSU rDNA Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.196, year: 2008

  19. Molecular characterization of Cryptosporidium spp. in poultry from Brazil

    Science.gov (United States)

    Cryptosporidiosis is an important zoonotic disease caused by the protozoa Cryptosporidium. Infections in birds are mainly caused by three species C. meleagridis, C. baileyi, and C. galli. C. meleagridis is the third most common cause of cryptosporidiosis in immunocompromised and immunocompetent huma...

  20. Cryptosporidium infection in infancy as a cause of malnutrition

    DEFF Research Database (Denmark)

    Mølbak, Kare; Andersen, M; Aaby, Peter

    1997-01-01

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

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

    NARCIS (Netherlands)

    Medema, Gerriet Jan

    1999-01-01

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

  2. Failed attempt of Cryptosporidium andersoni infection in lambs

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Ditrich, Oleg; Kouba, M.; Sak, Bohumil; Vítovec, J.; Květoňová, Dana

    2004-01-01

    Roč. 51, č. 4 (2004), s. 373-374 ISSN 0015-5683 R&D Projects: GA AV ČR IBS6022006 Institutional research plan: CEZ:AV0Z6022909 Keywords : cryptosporidiosis * Cryptosporidium andersoni * experimental infection Subject RIV: DJ - Water Pollution ; Quality Impact factor: 0.837, year: 2004

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

    Czech Academy of Sciences Publication Activity Database

    Laatamna, A.E.; Wágnerová, P.; Sak, Bohumil; Květoňová, Dana; Aissi, M.; Rost, M.; Kváč, Martin

    2013-01-01

    Roč. 197, 1-2 (2013), s. 350-353 ISSN 0304-4017 Grant - others:GAJU(CZ) 022/2010/Z; GAJU(CZ) 011/2013/Z Institutional support: RVO:60077344 Keywords : horses * Cryptosporidium hedgehog genotype * PCR * SSU * GP60 Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.545, year: 2013

  4. Cryptosporidium parvum and Isopora belli infections among patients ...

    African Journals Online (AJOL)

    Objective: To assess the importance of Cryptosporidium parvum and Isospora belli infections as a cause of diarrhoea among patients admitted to the Medical Wards in Queen Elizabeth Central Hospital (QECH) in Blantyre, Malawi. Design: Prospective case control study. Subjects: One hundred and twenty one patients with ...

  5. Prevalence of Cryptosporidium Species in Patients with Human ...

    African Journals Online (AJOL)

    Cryptosporidium is said to cause diarrhoea in HIV / AIDS patients. The study was done to determine the prevalence of cryptosporidiosis in Lagos. Stool samples were collected from 193 HIV positive and 200 HIV negative (control) patients presenting with diarrhoea at LUTH. The patient or a close relative filled a ...

  6. Eliminatie van virussen, Cryptosporidium en Giardia door drinkwaterzuiveringsprocessen

    NARCIS (Netherlands)

    Medema GJ; Theunissen J; MGB

    1996-01-01

    A study on the removal efficiency of drinking water treatment processes for viruses and protozoa (Cryptosporidium/Giardia). The description is based on the best available Dutch and, if data on the Dutch situation are absent, international research data. The approach is valid for well-designed and

  7. A Study of Cryptosporidium parvum Genotypes and Population Structure

    Directory of Open Access Journals (Sweden)

    G Widmer

    1998-09-01

    Full Text Available Genetic evidence for the occurrence of two Cryptosporidium parvum subgroups is presented. This evidence is based on restriction fragment length polymorphism analysis of several independent loci. Sequence analysis of the b -tubulin intron revealed additional polymorphism. The stability of the genetic profiles following passage of C. parvum isolates between different hosts was investigated.

  8. Age related susceptibility of pigs to Cryptosporidium scrofarum infection

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Němejc, K.; Kestřánová, M.; Květoňová, Dana; Wagnerová, Pavla; Kotková, Michaela; Rost, M.; Samková, E.; McEvoy, J.; Sak, Bohumil

    2014-01-01

    Roč. 202, 3-4 (2014), s. 330-334 ISSN 0304-4017 R&D Projects: GA MŠk(CZ) LH11061 Institutional support: RVO:60077344 Keywords : Cryptosporidium scrofarum * molecular analyses * transmission studies * susceptibility * infection * pigs Subject RIV: EE - Microbiology, Virology Impact factor: 2.460, year: 2014

  9. Prevalence and risk factors for Ascaris and Cryptosporidium ...

    African Journals Online (AJOL)

    Diseases in particular parasitic infestation is among the drawbacks to profitable pig production since parasites compromise the production and reproduction performance of infested pigs. The objective of this study was to estimate the prevalence and associated risk factors for Ascaris and Cryptosporidium infestations in pigs ...

  10. In vitro cultivation of Cryptosporidium parvum. A literature survey

    NARCIS (Netherlands)

    Schets FM; Leenen EJTM; MGB

    1998-01-01

    In vivo en in vitro modellen voor kweek van Cryptosporidium geven informatie over de infectiviteit van oocysten in ruw water en drinkwater en kunnen de identificatie van middelen met een anticryptosporidiele werking vereenvoudigen. In vivo modellen zijn echter duur, onpractisch en niet in elk

  11. Unrecognized Ingestion of Toxoplasma gondii Oocysts Leads to Congenital Toxoplasmosis and Causes Epidemics in North America

    Science.gov (United States)

    Boyer, Kenneth; Hill, Dolores; Mui, Ernest; Wroblewski, Kristen; Karrison, Theodore; Dubey, J. P.; Sautter, Mari; Noble, A. Gwendolyn; Withers, Shawn; Swisher, Charles; Heydemann, Peter; Hosten, Tiffany; Babiarz, Jane; Lee, Daniel

    2011-01-01

    (See the Editorial Commentary by Linn, on pages 1090–1.) Background. Congenital toxoplasmosis presents as severe, life-altering disease in North America. If mothers of infants with congenital toxoplasmosis could be identified by risks, it would provide strong support for educating pregnant women about risks, to eliminate this disease. Conversely, if not all risks are identifiable, undetectable risks are suggested. A new test detecting antibodies to sporozoites demonstrated that oocysts were the predominant source of Toxoplasma gondii infection in 4 North American epidemics and in mothers of children in the National Collaborative Chicago-based Congenital Toxoplasmosis Study (NCCCTS). This novel test offered the opportunity to determine whether risk factors or demographic characteristics could identify mothers infected with oocysts. Methods. Acutely infected mothers and their congenitally infected infants were evaluated, including in-person interviews concerning risks and evaluation of perinatal maternal serum samples. Results. Fifty-nine (78%) of 76 mothers of congenitally infected infants in NCCCTS had primary infection with oocysts. Only 49% of these mothers identified significant risk factors for sporozoite acquisition. Socioeconomic status, hometown size, maternal clinical presentations, and ethnicity were not reliable predictors. Conclusions. Undetected contamination of food and water by oocysts frequently causes human infections in North America. Risks are often unrecognized by those infected. Demographic characteristics did not identify oocyst infections. Thus, although education programs describing hygienic measures may be beneficial, they will not suffice to prevent the suffering and economic consequences associated with congenital toxoplasmosis. Only a vaccine or implementation of systematic serologic testing of pregnant women and newborns, followed by treatment, will prevent most congenital toxoplasmosis in North America. PMID:22021924

  12. Protecting chickens against coccidiosis in floor pens by administering Eimeria oocysts using gel beads or spray vaccination.

    Science.gov (United States)

    Jenkins, Mark C; Parker, Carolyn; O'Brien, Celia; Persyn, Joseph; Barlow, Darren; Miska, Katarzyna; Fetterer, Raymond

    2013-09-01

    Control of avian coccidiosis is increasingly being achieved by the administration of low doses of Eimeria oocysts to newly hatched chicks. The purpose of this study was to test the efficacy of gel beads containing a mixture of Eimeria acervulina, Eimeria maxima, and Eimeria tenella oocysts as a vaccine to protect broilers raised in contact with litter. Newly hatched chicks were either sprayed with an aqueous suspension of Eimeria oocysts or were allowed to ingest feed containing Eimeria oocysts-incorporated gel beads. Control, 1-day-old chicks were given an equivalent number of Eimeria oocysts (10(3) total) by oral gavage or received no vaccine (nonimmunized controls). All chicks were raised in floor-pen cages in direct contact with litter. At 4 wk of age, all chickens and a control nonimmunized group received a high-dose E. acervulina, E. maxima, and E. tenella challenge infection. Chickens immunized with Eimeria oocysts in gel beads or by spray vaccination displayed significantly (P 0.05) from chickens immunized by oral gavage or from nonimmunized, noninfected controls. Oocyst excretion after Eimeria challenge by all immunized groups was about 10-fold less than in nonimmunized controls. These findings indicate that immunization efficacy of gel beads and spray vaccination is improved by raising immunized chicks in contact with litter.

  13. Cryptosporidium species and Cryptosporidium parvum subtypes in dairy calves and goat kids reared under traditional farming systems in Turkey.

    Science.gov (United States)

    Taylan-Ozkan, Aysegul; Yasa-Duru, Sibel; Usluca, Selma; Lysen, Colleen; Ye, Jianbin; Roellig, Dawn M; Feng, Yaoyu; Xiao, Lihua

    2016-11-01

    Molecular characterizations of Cryptosporidium spp. in ruminants reared under traditional animal management systems are scarce and studies conducted thus far have revealed largely an absence of the pathogenic and zoonotic species Cryptosporidium parvum in pre-weaned animals. In this study, we examined Cryptosporidium species and subtype distribution in free-range pre-weaned dairy calves and goat kids with diarrhea. Cryptosporidium-positive specimens from pre-weaned calves on 10 farms and goat kids on 4 farms in Ankara, Balikesir, Corum, Kirikkale, and Kirsehir Provinces, Turkey were genotyped by PCR-restriction length polymorphism analysis of the small subunit rRNA gene, which identified C. parvum in 27 calves and 9 goat kids and Cryptosporidium ryanae in 1 calf. Among the C. parvum isolates successfully subtyped by DNA sequence analysis of the 60 kDa glycoprotein gene, three subtypes were detected in calves, including IIaA13G2R1 (20/23), IIdA18G1 (2/23), and IIdA20G1b (1/23), and four subtypes were detected in goat kids, including IIaA13G2R1 (3/8), IIaA15G1R1 (2/8), IIdA22G1 (2/8), and IIdA18G1 (1/8). Data of the study suggest that dairy calves reared in a traditional cow-calf system in Turkey are mainly infected with a C. parvum subtype rarely seen elsewhere, whereas goat kids are infected with diverse subtypes. As all five C. parvum subtypes found in this study are known human pathogens, pre-weaned farm animals could play a potential role in the transmission of human cryptosporidiosis. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. RNA-Seq analysis during the life cycle of Cryptosporidium parvum reveals significant differential gene expression between proliferating stages in the intestine and infectious sporozoites.

    Science.gov (United States)

    Lippuner, Christoph; Ramakrishnan, Chandra; Basso, Walter U; Schmid, Marc W; Okoniewski, Michal; Smith, Nicholas C; Hässig, Michael; Deplazes, Peter; Hehl, Adrian B

    2018-05-01

    Cryptosporidium parvum is a major cause of diarrhoea in humans and animals. There are no vaccines and few drugs available to control C. parvum. In this study, we used RNA-Seq to compare gene expression in sporozoites and intracellular stages of C. parvum to identify genes likely to be important for successful completion of the parasite's life cycle and, thereby, possible targets for drugs or vaccines. We identified 3774 protein-encoding transcripts in C. parvum. Applying a stringent cut-off of eight fold for determination of differential expression, we identified 173 genes (26 coding for predicted secreted proteins) upregulated in sporozoites. On the other hand, expression of 1259 genes was upregulated in intestinal stages (merozoites/gamonts) with a gene ontology enrichment for 63 biological processes and upregulation of 117 genes in 23 metabolic pathways. There was no clear stage specificity of expression of AP2-domain containing transcription factors, although sporozoites had a relatively small repertoire of these important regulators. Our RNA-Seq analysis revealed a new calcium-dependent protein kinase, bringing the total number of known calcium-dependent protein kinases (CDPKs) in C. parvum to 11. One of these, CDPK1, was expressed in all stages, strengthening the notion that it is a valid drug target. By comparing parasites grown in vivo (which produce bona fide thick-walled oocysts) and in vitro (which are arrested in sexual development prior to oocyst generation) we were able to confirm that genes encoding oocyst wall proteins are expressed in gametocytes and that the proteins are stockpiled rather than generated de novo in zygotes. RNA-Seq analysis of C. parvum revealed genes expressed in a stage-specific manner and others whose expression is required at all stages of development. The functional significance of these can now be addressed through recent advances in transgenics for C. parvum, and may lead to the identification of viable drug and vaccine

  15. In vitro evaluation of the disinfection efficacy on Eimeria tenella unsporulated oocysts isolated from broilers.

    Science.gov (United States)

    Guimarães, José S; Bogado, Alexey L Gomel; da Cunha, Thiago Cezar B; Garcia, João Luis

    2007-01-01

    The objective of this study was to evaluate in vitro the action of eight chemical principles by disinfection efficacy (DE) of Eimeria tenella oocysts. Disinfection efficacy was evaluated by either destruction or sporulation inhibition of the oocysts. Eight treatments were performed: T1 (Glutaraldehyde 42.5 g + Benzalkonium Chloride 7.5 g); T2 (Benzalkonium chloride + quaternary ammonium salt); T3 (formol 37% + Sodium Dodecylbenzene Sulfonate 12%); T4 (sodium hypochlorite 2%); T5 (Orthodichlorobenzene 60% + Xylene 30%); T6 (Polyoctyl polyamino ethyl glycine + Polyoxyethylene alkylphenol ether + Sodium Chloride); T7 (Chloramine T) and finally T8 (free iodine 2.25% + Phosphoric acid 15 g). The control test was carried out with distilled water (T9). The best DE were observed, respectively, in T3 (79.49%), T5 (75.60%) and T4 (65.56%) treatments.

  16. Immunoproteomic analysis of the protein repertoire of unsporulated Eimeria tenella oocysts

    Directory of Open Access Journals (Sweden)

    Zhang Zhenchao

    2017-01-01

    Full Text Available The apicomplexan protozoans Eimeria spp. cause coccidioses, the most common intestinal diseases in chickens. Coccidiosis is associated with significant animal welfare issues and has a high economic impact on the poultry industry. Lack of a full understanding of immunogenic molecules and their precise functions involved in the Eimeria life cycles may limit development of effective vaccines and drug therapies. In this study, immunoproteomic approaches were used to define the antigenic protein repertoire from the total proteins of unsporulated Eimeria tenella oocysts. Approximately 101 protein spots were recognized in sera from chickens infected experimentally with E. tenella. Forty-six spots of unsporulated oocysts were excised from preparative gels and identified by matrix-assisted laser desorption ionization time-of-flight MS (MALDI-TOF-MS and MALDI-TOF/TOF-MS. For unsporulated oocysts, 13 known proteins of E. tenella and 17 homologous proteins to other apicomplexan or protozoan parasites were identified using the ‘Mascot’ server. The remaining proteins were searched against the E. tenella protein sequence database using the ‘Mascot in-house’ search engine (version 2.1 in automated mode, and 12 unknown proteins were identified. The amino acid sequences of the unknown proteins were searched using BLAST against non-redundant sequence databases (NCBI, and 9 homologous proteins in unsporulated oocyst were found homologous to proteins of other apicomplexan parasites. These findings may provide useful evidence for understanding parasite biology, pathogenesis, immunogenicity and immune evasion mechanisms of E. tenella.

  17. Oocysts of Hepatozoon canis in Rhipicephalus (Boophilus) microplus collected from a naturally infected dog.

    Science.gov (United States)

    de Miranda, Renata Lima; de Castro, Jacqueline Ribeiro; Olegário, Maria Marlene Martins; Beletti, Marcelo Emílio; Mundim, Antonio Vicente; O'Dwyer, Lucia Helena; Eyal, Osnat; Talmi-Frank, Dalit; Cury, Márcia Cristina; Baneth, Gad

    2011-05-11

    Canine hepatozoonosis is a tick-borne disease caused by protozoans of the genus Hepatozoon. Several tick species have been implicated as potential vectors. Therefore, extensive studies are needed to determine the 'natural' endemic cycle of this parasite. This paper presents the first report of the presence of Hepatozoon canis oocysts in Rhipicephalus (Boophilus) microplus collected from an infected dog. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. Toxoplasma gondii: infection natural congenital in cattle and an experimental inoculation of gestating cows with oocysts.

    Science.gov (United States)

    Costa, Gustavo Henrique Nogueira; da Costa, Alvimar José; Lopes, Welber Daniel Zanetti; Bresciani, Katia Denise Saraiva; dos Santos, Thais Rabelo; Esper, César Roberto; Santana, Aureo Evangelista

    2011-01-01

    Two studies, of a natural infection and an experimental infection, were performed in order to study congenital transmission of Toxoplasma gondii in cattle. In the first study, 50 fetuses were harvested from gestating cows that were eutanasied at a municipal slaughterhouse in Jaboticabal, São Paulo state, Brazil. In the second study, 11 gestating cows were divided into four groups for inoculation with T. gondii: GI consisted of three cows inoculated with 1.0 × 10(5) oocysts during their first trimester of gestation; GII consisted of three cows inoculated with 1.0 × 10(5) oocysts during their second trimester of gestation; GIII consisted of three cows inoculated with 1.0 × 10(5) oocysts during their last trimester of gestation; and GIV consisted of two control cows, one during its first and the other during its second trimester of gestation. In both studies, the presence of T. gondii was confirmed both indirectly by immunofluorescence assay (IFAT). In the natural infection experiment, 18% (9/50) of the gestating cows were confirmed to have specific antibodies (IFAT--1:64) against T. gondii. The bioassay was able to diagnose the presence of T. gondii in the tissue samples from three calves. In the second experiment, the nine cows from groups I, II and III presented with specific antibodies (IFAT) against T. gondii. In contrast, T. gondii could not be detected by IFAT, histopathological examination or the bioassay in any of the nine calves born to cows experimentally infected with T. gondii oocysts. Based on the results from both studies, we conclude that congenital infection of T. gondii in cattle, while infrequent, does occur naturally. The pathogenicity of the strain of T. gondii may influence the likelihood of this route of transmission. Copyright © 2010 Elsevier Inc. All rights reserved.

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

    NARCIS (Netherlands)

    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

  20. Probabilistic quantitative microbial risk assessment model of farmer exposure to Cryptosporidium spp. in irrigation water within Kumasi Metropolis-Ghana

    DEFF Research Database (Denmark)

    Sampson, Angelina; Owusu-Ansah, Emmanuel de-Graft Johnson; Mills-Robertson, Felix C.

    2017-01-01

    causing gastroenteritis. The results indicate high positive levels of Cryptosporidium in the irrigation water, however, the levels of Cryptosporidium decreases during the rainfall seasons, risk assessment results show that, farmers face a higher risk of being infected by Cryptosporidium due to frequent...

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

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; McEvoy, J.; Loudová, M.; Stenger, B.; Sak, Bohumil; Květoňová, Dana; Ditrich, Oleg; Rašková, Veronika; Moriarty, E.; Rost, M.; Macholán, Miloš; Piálek, Jaroslav

    2013-01-01

    Roč. 43, č. 10 (2013), s. 805-817 ISSN 0020-7519 R&D Projects: GA ČR GA206/08/0640; GA MŠk(CZ) LH11061 Institutional support: RVO:60077344 ; RVO:67985904 ; RVO:68081766 Keywords : Cryptosporidium tyzzeri * house mouse * hybrid zone * coevolution Subject RIV: EG - Zoology; GJ - Animal Vermins ; Diseases, Veterinary Medicine (BC-A) Impact factor: 3.404, year: 2013

  2. Natural infection of Cryptosporidium muris (Apicomplexa: Cryptosporiidae) in Siberian chipmunks

    Czech Academy of Sciences Publication Activity Database

    Hůrková, L.; Hajdušek, Ondřej; Modrý, David

    2003-01-01

    Roč. 39, č. 2 (2003), s. 441ů444 ISSN 0090-3558 Grant - others:GA FRVŠ(CZ) 1260/2001 Institutional research plan: CEZ:AV0Z6022909; CEZ:MSM 123100003; CEZ:MSM 161700001 Keywords : BALB/c mice * Cryptosporidium muris * Eutamias sibiricus Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 0.793, year: 2003

  3. Cryptosporidium muris in a Reticulated Giraffe (Giraffa camelopardalis reticulata)

    Czech Academy of Sciences Publication Activity Database

    Kodádková, A.; Kváč, M.; Ditrich, Oleg; Sak, Bohumil; Xiao, L.

    2010-01-01

    Roč. 96, č. 1 (2010), s. 211-212 ISSN 0022-3395 R&D Projects: GA ČR GA524/05/0992; GA ČR GP523/07/P117 Institutional research plan: CEZ:AV0Z60220518 Keywords : Cryptosporidium muris * Reticulated giraffe * natural infection Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 1.208, year: 2010

  4. Cryptosporidium and Giardia in Swimming Pools, Atlanta, Georgia

    Centers for Disease Control (CDC) Podcasts

    In this podcast, Dan Rutz speaks with Dr. Joan Shields, a guest researcher with the Healthy Swimming Program at CDC, about an article in June 2008 issue of Emerging Infectious Diseases reporting on the results of a test of swimming pools in the greater Atlanta, Georgia area. Dr. Shields tested 160 pools in metro Atlanta last year for Cryptosporidium and Giardia. These germs cause most recreational water associated outbreaks.

  5. La confusa taxonomía de Cryptosporidium

    Directory of Open Access Journals (Sweden)

    Gregorio Pérez-Cordón

    2006-10-01

    Full Text Available Los últimos descubrimientos en la biología y filogenética de Cryptosporidium refuerzan la necesidad de una exhaustiva revisión del ciclo de vida y la taxonomía de este parásito. Tanto futuros estudios de cultivo in vitro e in vivo así como estudios moleculares y genéticos permitirán avanzar en el profundo conocimiento de este interesante parásito.

  6. Native and introduced squirrels in Italy host different Cryptosporidium spp.

    Czech Academy of Sciences Publication Activity Database

    Prediger, Jitka; Horčičková, Michaela; Hofmannová, L.; Sak, Bohumil; Ferrari, N.; Mazzamuto, M.V.; Romeo, C.; Wauters, L.A.; McEvoy, J.; Kváč, Martin

    2017-01-01

    Roč. 61, OCT (2017), s. 64-75 ISSN 0932-4739 R&D Projects: GA ČR GA15-01090S; GA MŠk LTAUSA17165 Institutional support: RVO:60077344 Keywords : Cryptosporidium * gp60 * infection * Italy * phylogeny * tree squirrels Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine OBOR OECD: Veterinary science Impact factor: 2.581, year: 2016

  7. Plasmodium P-Type Cyclin CYC3 Modulates Endomitotic Growth during Oocyst Development in Mosquitoes.

    Science.gov (United States)

    Roques, Magali; Wall, Richard J; Douglass, Alexander P; Ramaprasad, Abhinay; Ferguson, David J P; Kaindama, Mbinda L; Brusini, Lorenzo; Joshi, Nimitray; Rchiad, Zineb; Brady, Declan; Guttery, David S; Wheatley, Sally P; Yamano, Hiroyuki; Holder, Anthony A; Pain, Arnab; Wickstead, Bill; Tewari, Rita

    2015-11-01

    Cell-cycle progression and cell division in eukaryotes are governed in part by the cyclin family and their regulation of cyclin-dependent kinases (CDKs). Cyclins are very well characterised in model systems such as yeast and human cells, but surprisingly little is known about their number and role in Plasmodium, the unicellular protozoan parasite that causes malaria. Malaria parasite cell division and proliferation differs from that of many eukaryotes. During its life cycle it undergoes two types of mitosis: endomitosis in asexual stages and an extremely rapid mitotic process during male gametogenesis. Both schizogony (producing merozoites) in host liver and red blood cells, and sporogony (producing sporozoites) in the mosquito vector, are endomitotic with repeated nuclear replication, without chromosome condensation, before cell division. The role of specific cyclins during Plasmodium cell proliferation was unknown. We show here that the Plasmodium genome contains only three cyclin genes, representing an unusual repertoire of cyclin classes. Expression and reverse genetic analyses of the single Plant (P)-type cyclin, CYC3, in the rodent malaria parasite, Plasmodium berghei, revealed a cytoplasmic and nuclear location of the GFP-tagged protein throughout the lifecycle. Deletion of cyc3 resulted in defects in size, number and growth of oocysts, with abnormalities in budding and sporozoite formation. Furthermore, global transcript analysis of the cyc3-deleted and wild type parasites at gametocyte and ookinete stages identified differentially expressed genes required for signalling, invasion and oocyst development. Collectively these data suggest that cyc3 modulates oocyst endomitotic development in Plasmodium berghei.

  8. Detection of Toxoplasma gondii oocysts in different water resources by Loop Mediated Isothermal Amplification (LAMP).

    Science.gov (United States)

    Gallas-Lindemann, Carmen; Sotiriadou, Isaia; Mahmoodi, Mohammad Reza; Karanis, Panagiotis

    2013-02-01

    Human toxoplasmosis is potentially contracted due to consumption of contaminated drinking water and represents an increasing public health risk worldwide. Toxoplasma gondii oocysts can be resistant to standard disinfection processes, including UV radiation. Increased awareness of the risk of waterborne toxoplasmosis outbreaks has led to an increase in research interest in the detection of oocysts in environmental water systems. Ninety-five environmental water samples from the Lower Rhine area in Germany have been included in the study and examined for the presence of Toxoplasma. Water samples were filtered or flocculated by aluminum sulfate and purified by sucrose density gradient. DNA was then extracted, and the DNA samples were then examined by LAMP analysis. T. gondii DNA was detected in eight out of 83 (9.6%) influent and effluent samples obtained from wastewater treatment plants. All samples (n=12) from the surface, ground, raw and tap waters tested negative. The purpose of this work was to investigate the occurrence and distribution of Toxoplasma oocysts on the Lower Rhine in Germany. Our study provides evidence that the assay is a sensitive, specific, rapid and cost effective method for the detection of T. gondii and is useful for both the investigations of cases of waterborne outbreaks and for identifying the source of contamination. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. Plasmodium P-Type Cyclin CYC3 Modulates Endomitotic Growth during Oocyst Development in Mosquitoes

    KAUST Repository

    Roques, Magali; Wall, Richard J.; Douglass, Alexander P.; Ramaprasad, Abhinay; Ferguson, David J. P.; Kaindama, Mbinda L.; Brusini, Lorenzo; Joshi, Nimitray; Rchiad, ‍ Zineb; Brady, Declan; Guttery, David S.; Wheatley, Sally P.; Yamano, Hiroyuki; Holder, Anthony A.; Pain, Arnab; Wickstead, Bill; Tewari, Rita

    2015-01-01

    Cell-cycle progression and cell division in eukaryotes are governed in part by the cyclin family and their regulation of cyclin-dependent kinases (CDKs). Cyclins are very well characterised in model systems such as yeast and human cells, but surprisingly little is known about their number and role in Plasmodium, the unicellular protozoan parasite that causes malaria. Malaria parasite cell division and proliferation differs from that of many eukaryotes. During its life cycle it undergoes two types of mitosis: endomitosis in asexual stages and an extremely rapid mitotic process during male gametogenesis. Both schizogony (producing merozoites) in host liver and red blood cells, and sporogony (producing sporozoites) in the mosquito vector, are endomitotic with repeated nuclear replication, without chromosome condensation, before cell division. The role of specific cyclins during Plasmodium cell proliferation was unknown. We show here that the Plasmodium genome contains only three cyclin genes, representing an unusual repertoire of cyclin classes. Expression and reverse genetic analyses of the single Plant (P)-type cyclin, CYC3, in the rodent malaria parasite, Plasmodium berghei, revealed a cytoplasmic and nuclear location of the GFP-tagged protein throughout the lifecycle. Deletion of cyc3 resulted in defects in size, number and growth of oocysts, with abnormalities in budding and sporozoite formation. Furthermore, global transcript analysis of the cyc3-deleted and wild type parasites at gametocyte and ookinete stages identified differentially expressed genes required for signalling, invasion and oocyst development. Collectively these data suggest that cyc3 modulates oocyst endomitotic development in Plasmodium berghei.

  10. Plasmodium P-Type Cyclin CYC3 Modulates Endomitotic Growth during Oocyst Development in Mosquitoes

    KAUST Repository

    Roques, Magali

    2015-11-13

    Cell-cycle progression and cell division in eukaryotes are governed in part by the cyclin family and their regulation of cyclin-dependent kinases (CDKs). Cyclins are very well characterised in model systems such as yeast and human cells, but surprisingly little is known about their number and role in Plasmodium, the unicellular protozoan parasite that causes malaria. Malaria parasite cell division and proliferation differs from that of many eukaryotes. During its life cycle it undergoes two types of mitosis: endomitosis in asexual stages and an extremely rapid mitotic process during male gametogenesis. Both schizogony (producing merozoites) in host liver and red blood cells, and sporogony (producing sporozoites) in the mosquito vector, are endomitotic with repeated nuclear replication, without chromosome condensation, before cell division. The role of specific cyclins during Plasmodium cell proliferation was unknown. We show here that the Plasmodium genome contains only three cyclin genes, representing an unusual repertoire of cyclin classes. Expression and reverse genetic analyses of the single Plant (P)-type cyclin, CYC3, in the rodent malaria parasite, Plasmodium berghei, revealed a cytoplasmic and nuclear location of the GFP-tagged protein throughout the lifecycle. Deletion of cyc3 resulted in defects in size, number and growth of oocysts, with abnormalities in budding and sporozoite formation. Furthermore, global transcript analysis of the cyc3-deleted and wild type parasites at gametocyte and ookinete stages identified differentially expressed genes required for signalling, invasion and oocyst development. Collectively these data suggest that cyc3 modulates oocyst endomitotic development in Plasmodium berghei.

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

    Science.gov (United States)

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

    2014-01-01

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

  12. Management of a Cryptosporidium hominis Outbreak in a Day- care Center

    NARCIS (Netherlands)

    Vandenberg, Olivier; Robberecht, Françoise; Dauby, Nicolas; Moens, Catherine; Talabani, Hana; Dupont, Eddy; Menotti, Jean; van Gool, Tom; Levy, Jack

    2012-01-01

    Background: Cryptosporidium outbreaks in day-care centers (DCCs) occur commonly. However, controlling spread of infection in these settings is difficult, and data about effectiveness of different control strategies are sparse. In this study, a Cryptosporidium outbreak in a large DCC located in

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

    Directory of Open Access Journals (Sweden)

    Sahatchai Tangtrongsup

    2017-05-01

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

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

    Science.gov (United States)

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

    2017-05-10

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

  15. Cryptosporidium scrofarum n. sp. (Apicomplexa: Cryptosporidiidae) in domestic pigs (Sus scrofa)

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Kestřánová, M.; Pinková, Martina; Květoňová, Dana; Kalinová, Jana; Wagnerová, Pavla; Kotková, Michaela; Vitovec, J.; Ditrich, Oleg; McEvoy, J.; Stenger, B.; Sak, Bohumil

    2013-01-01

    Roč. 191, 3-4 (2013), s. 218-227 ISSN 0304-4017 R&D Projects: GA MŠk(CZ) LH11061 Institutional support: RVO:60077344 ; RVO:67985904 Keywords : Cryptosporidium scrofarum * Taxonomy * Morphology * Molecular analyses * Transmission studies * Cryptosporidium pig genotype II Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.545, year: 2013

  16. Impacts of population growth, urbanisation and sanitation changes on global human Cryptosporidium emissions to surface water

    NARCIS (Netherlands)

    Hofstra, Nynke; Vermeulen-Henstra, Lucie

    2016-01-01

    Cryptosporidium is a pathogenic protozoan parasite and is a leading cause of diarrhoea worldwide. The concentration of Cryptosporidium in the surface water is a determinant for probability of exposure and the risk of disease. Surface water concentrations are expected to change with population

  17. Chlorine Dioxide Disinfection in the Use of Individual Water Purification Devices

    Science.gov (United States)

    2006-03-01

    CTs ranging from 1.7-17.6 mg-min/L necessary for 2-log Giardia muris cyst inactivation (reference 23). The SWTR provides the following CT values...reference 3). A comparison of CTs required for a 2-log inactivation for E. Coli bacteria, Poliovirus 1, and Giardia cysts showed Giardia cysts were 2-5...Cryptosporidium oocysts are the most resistant, being 8-16 times more resistant than Giardia cysts (reference 5). Chlorine dioxide’s general disinfection

  18. Detection of Toxoplasma gondii oocysts in soils in northwestern China using a new semi-nested PCR assay.

    Science.gov (United States)

    Wang, Meng; Meng, Peng; Ye, Qiang; Pu, Yuan-Hua; Yang, Xiao-Yu; Luo, Jian-Xun; Zhang, Nian-Zhang; Zhang, De-Lin

    2014-09-28

    Toxoplasma gondii is a zoonotic pathogen that can infect a range of animals and humans. Ingestion of T. gondii oocysts in soil is a significant transmission route for humans and animals acquiring toxoplasmosis. In the present study, we developed a new semi-nested PCR method to determine T. gondii oocysts distribution in soils in northwestern China. The one tube semi-nested PCR assay was developed to detect the oocysts of T. gondii in soil, targeting the repetitive 529 bp fragment of T. gondii genomic DNA. Then a total of 268 soil samples, including 148 samples from Gansu Province and 120 samples from Qinghai Province, northwestern China, were examined by the semi-nested PCR method. One third of the positive samples were sequenced. The sensitivity of the semi-nested PCR assay was 10(2)  T. gondii oocysts in 5 g soil sample. Investigation of soil samples from northwestern China showed that 34 out of 268 soil samples (12.69%) were T. gondii positive. Sequences of the partial 529 bp fragments varied from 0-1.2% among the sequenced samples. The prevalence of T. gondii oocysts in soil from cities (24/163) was slightly higher than that in soils from pasturing areas (10/105) (P = 0.21). Among the different regions in cities, the prevalence of T. gondii oocysts in soils from parks was 14.15%, whereas that in soils from schools was 19.05%. The present study firstly reported the prevalence of T. gondii oocysts in soils in northwest China using a novel semi-nested PCR assay, which provided baseline data for the effective prevention and control of toxoplasmosis in this region.

  19. Pathogen inactivation techniques.

    Science.gov (United States)

    Pelletier, J P R; Transue, S; Snyder, E L

    2006-01-01

    The desire to rid the blood supply of pathogens of all types has led to the development of many technologies aimed at the same goal--eradication of the pathogen(s) without harming the blood cells or generating toxic chemical agents. This is a very ambitious goal, and one that has yet to be achieved. One approach is to shun the 'one size fits all' concept and to target pathogen-reduction agents at the Individual component types. This permits the development of technologies that might be compatible with, for example, plasma products but that would be cytocidal and thus incompatible with platelet concentrates or red blood cell units. The technologies to be discussed include solvent detergent and methylene blue treatments--designed to inactivate plasma components and derivatives; psoralens (S-59--amotosalen) designed to pathogen-reduce units of platelets; and two products aimed at red blood cells, S-303 (a Frale--frangible anchor-linker effector compound) and Inactine (a binary ethyleneimine). A final pathogen-reduction material that might actually allow one material to inactivate all three blood components--riboflavin (vitamin B2)--is also under development. The sites of action of the amotosalen (S-59), the S-303 Frale, Inactine, and riboflavin are all localized in the nucleic acid part of the pathogen. Solvent detergent materials act by dissolving the plasma envelope, thus compromising the integrity of the pathogen membrane and rendering it non-infectious. By disrupting the pathogen's ability to replicate or survive, its infectivity is removed. The degree to which bacteria and viruses are affected by a particular pathogen-reducing technology relates to its Gram-positive or Gram-negative status, to the sporulation characteristics for bacteria, and the presence of lipid or protein envelopes for viruses. Concerns related to photoproducts and other breakdown products of these technologies remain, and the toxicology of pathogen-reduction treatments is a major ongoing area

  20. Free radical inactivation of trypsin

    International Nuclear Information System (INIS)

    Cudina, Ivana; Jovanovic, S.V.

    1988-01-01

    Reactivities of free radical oxidants, radical OH, Br2-anion radical and Cl 3 COO radical and a reductant, CO2-anion radical, with trypsin and reactive protein components were determined by pulse radiolysis of aqueous solutions at pH 7, 20 0 C. Highly reactive free radicals, radical OH, Br2-anion radical and CO2-anion radical, react with trypsin at diffusion controlled rates. Moderately reactive trichloroperoxy radical, k(Cl 3 COO radical + trypsin) preferentially oxidizes histidine residues. The efficiency of inactivation of trypsin by free radicals is inversely proportional to their reactivity. The yields of inactivation of trypsin by radical OH, Br2-anion radical and CO2-anion radical are low, G(inactivation) = 0.6-0.8, which corresponds to ∼ 10% of the initially produced radicals. In contrast, Cl 3 COO radical inactivates trypsin with ∼ 50% efficiency, i.e. G(inactivation) = 3.2. (author)

  1. rROP2 from Toxoplasma gondii as a potential vaccine against oocyst shedding in domestic cats

    Directory of Open Access Journals (Sweden)

    Dauton Luiz Zulpo

    Full Text Available Abstract The aim of the present study was to evaluate oocyst shedding in cats immunized by nasal route with T. gondii proteins ROP2. Twelve short hair cats (Felis catus were divided in three groups G1, G2 and G3 (n=4. Animals from G1 received 100 μg of rROP2 proteins plus 20 μg of Quil-A, G2 received 100 μg of BSA plus 20 μg of Quil-A, and the G3 only saline solution (control group. All treatments were done by intranasal route at days 0, 21, 42, and 63. The challenge was performed in all groups on day 70 with ≅ 800 tissue cysts of ME-49 strain by oral route. Animals from G1 shed less oocysts (86.7% than control groups. ELISA was used to detect anti-rROP2 IgG and IgA, however, there were no correlation between number of oocyst shedding by either IgG or IgA antibody levels. In the present work, in spite of lesser oocysts production in immunized group than control groups, it was not possible to associate the use of rROP2 via nostrils with protection against oocyst shedding. For the future, the use of either other recombinant proteins or DNA vaccine, in combination with rROP2 could be tested to try improving the efficacy of this kind of vaccine.

  2. Inactivation of Microorganisms

    Science.gov (United States)

    Alzamora, Stella Maris; Guerrero, Sandra N.; Schenk, Marcela; Raffellini, Silvia; López-Malo, Aurelio

    Minimal processing techniques for food preservation allow better retention of product flavor, texture, color, and nutrient content than comparable conventional treatments. A wide range of novel alternative physical factors have been intensely investigated in the last two decades. These physical factors can cause inactivation of microorganisms at ambient or sublethal temperatures (e.g., high hydrostatic pressure, pulsed electric fields, ultrasound, pulsed light, and ultraviolet light). These technologies have been reported to reduce microorganism population in foods while avoiding the deleterious effects of severe heating on quality. Among technologies, high-energy ultrasound (i.e., intensities higher than 1 W/cm2, frequencies between 18 and 100 kHz) has attracted considerable interest for food preservation applications (Mason et al., 1996; Povey and Mason, 1998).

  3. Mean inactivation dose (D)

    International Nuclear Information System (INIS)

    Vijayakumar, S.; Ng, T.C.; Raudkivi, U.; Meaney, T.J.

    1990-01-01

    By predicting treatment outcome to radiotherapy from in vitro radiobiological parameters, not only individual patient treatments can be tailored, but also new promising treatment protocols can be tried in patients in whom unfavorable outcome is predicted. In this respect, choosing the right parameter can be very important. Unlike D 0 and N which provide information of the distal part of the survival curve, mean inactivation dose (D) estimates overall radiosensitivity. However, the parameters reflecting the response at the clinically relevant low-dose region are neglected in the literature. In a literature survey of 98 papers in which survival curves or D 0 /N were used, only in 2 D was used. In 21 papers the D 0 /n values were important in drawing conclusions. By calculating D in 3 of these 21 papers, we show that the conclusion drawn may be altered with the use of D. The importance of ''low-dose-region-parameters'' is reviewed. (orig.)

  4. Plasmodium P-Type Cyclin CYC3 Modulates Endomitotic Growth during Oocyst Development in Mosquitoes

    Science.gov (United States)

    Ferguson, David J. P.; Kaindama, Mbinda L.; Brusini, Lorenzo; Joshi, Nimitray; Rchiad, Zineb; Brady, Declan; Guttery, David S.; Wheatley, Sally P.; Yamano, Hiroyuki; Holder, Anthony A.; Pain, Arnab; Wickstead, Bill; Tewari, Rita

    2015-01-01

    Cell-cycle progression and cell division in eukaryotes are governed in part by the cyclin family and their regulation of cyclin-dependent kinases (CDKs). Cyclins are very well characterised in model systems such as yeast and human cells, but surprisingly little is known about their number and role in Plasmodium, the unicellular protozoan parasite that causes malaria. Malaria parasite cell division and proliferation differs from that of many eukaryotes. During its life cycle it undergoes two types of mitosis: endomitosis in asexual stages and an extremely rapid mitotic process during male gametogenesis. Both schizogony (producing merozoites) in host liver and red blood cells, and sporogony (producing sporozoites) in the mosquito vector, are endomitotic with repeated nuclear replication, without chromosome condensation, before cell division. The role of specific cyclins during Plasmodium cell proliferation was unknown. We show here that the Plasmodium genome contains only three cyclin genes, representing an unusual repertoire of cyclin classes. Expression and reverse genetic analyses of the single Plant (P)-type cyclin, CYC3, in the rodent malaria parasite, Plasmodium berghei, revealed a cytoplasmic and nuclear location of the GFP-tagged protein throughout the lifecycle. Deletion of cyc3 resulted in defects in size, number and growth of oocysts, with abnormalities in budding and sporozoite formation. Furthermore, global transcript analysis of the cyc3-deleted and wild type parasites at gametocyte and ookinete stages identified differentially expressed genes required for signalling, invasion and oocyst development. Collectively these data suggest that cyc3 modulates oocyst endomitotic development in Plasmodium berghei. PMID:26565797

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

    Science.gov (United States)

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

    2016-05-01

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

  6. Cryptosporidium and Giardia in Swimming Pools, Atlanta, Georgia

    Centers for Disease Control (CDC) Podcasts

    2008-05-29

    In this podcast, Dan Rutz speaks with Dr. Joan Shields, a guest researcher with the Healthy Swimming Program at CDC, about an article in June 2008 issue of Emerging Infectious Diseases reporting on the results of a test of swimming pools in the greater Atlanta, Georgia area. Dr. Shields tested 160 pools in metro Atlanta last year for Cryptosporidium and Giardia. These germs cause most recreational water associated outbreaks.  Created: 5/29/2008 by Emerging Infectious Diseases.   Date Released: 5/29/2008.

  7. Prevalência de Cryptosporidium serpentis em serpentes de cativeiro Prevalence of Cryptosporidium serpentis in captive snakes

    Directory of Open Access Journals (Sweden)

    Paulo Custório Ruggiero

    2011-11-01

    Full Text Available Cryptosporidium é um protozoário encontrado em uma grande variedade de espécies animais como responsável por casos de gastrite e enterite, porém com epidemiologia pouco conhecida em animais silvestres. A presente investigação teve como objetivo avaliar a prevalência de Cryptosporidium serpentis em lavado gástrico de serpentes mantidas em cativeiro no serpentário do Instituto Butantan (São Paulo, Brasil. A coleta foi realizada uma semana após alimentação, evitando, assim, a regurgitação devido à manipulação. Foram realizados esfregaços do sedimento do lavado gástrico, obtido por centrifugação, corados pela técnica de coloração de Kinyoun. Parte do sedimento foi submetido à técnica de RFLP-PCR para identificação da espécie de Cryptosporidium. O serpentário é dividido em três seções, por espécie - a primeira com oito jibóias (Boa constrictor amarali, a segunda com dez jararacas (Bothropoides jararaca e a última com sete cascavéis (Caudisona durissa. A prevalência de C. serpentis encontrada neste estudo para as serpentes C. durissa, B. jararaca e Boa c. amarali, foi de 57,14% (04/07, 40% (04/10 e 37,5% (03/08, respectivamente, revelando importante ocorrência desse protozoário em serpentes de cativeiro. Apesar da alta prevalência encontrada, apenas as jiboias apresentaram sintomas como perda de peso e regurgitação, refletindo uma sensibilidade diferente da espécie para C. serpentis.Cryptosporidium is a protozoan found in a wide variety of animal species which is responsible for gastritis and enteritis, but its epidemiology is poorly known in wild animals. The present investigation aimed to evaluate the prevalence of Cryptosporidium serpentis in gastric aspirate of captive snakes from the public serpentarium of the Butantan Institute (São Paulo, Brazil. Sampling was performed preferably one week after feeding, thereby preventing regurgitation due to manipulation. Smears were done from the gastric

  8. Clinical Manifestations of Cryptosporidiosis and Identification of a New Cryptosporidium Subtype in Patients From Sonora, Mexico.

    Science.gov (United States)

    Urrea-Quezada, Alejandro; González-Díaz, Mariana; Villegas-Gómez, Isaac; Durazo, María; Hernández, Jesús; Xiao, Lihua; Valenzuela, Olivia

    2018-05-01

    The aim of this study was to identify the clinical manifestations of cryptosporidiosis and the distribution of Cryptosporidium spp. and subtypes in children in Sonora, Mexico. Two subtypes of C. parvum, including IIaA15G2R1 and IIcA5G3a, and 6 subtypes of Cryptosporidium hominis, including IaA14R3, IaA15R3, IbA12G3, IdA23, IeA11G3T3, and a new subtype IaA14R11, were identified. Cryptosporidium as an etiologic agent for acute gastroenteritis is discussed.

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

    Science.gov (United States)

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

    2018-02-01

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

  10. Experimental Infection with Sporulated Oocysts of Eimeria maxima (Apicomplexa: Eimeriidae) in Broiler

    Science.gov (United States)

    Brito, Luciana da S.; Pereira, Elder N.; da Silva, Augusta A.; Bentivóglio Costa Silva, Vinícius; Freitas, Fagner L. da C.

    2014-01-01

    Through this study we assessed the metabolic and pathological changes in broilers experimentally infected with oocysts of Eimeria maxima. To perform the experiment, we used 150 broiler strain cooB males, with ten days of age, were randomized according to weight and randomly assigned to two experimental groups: the control group was inoculated with 0.5 mL of distilled water; the infected group inoculated with 0.5 mL of solution containing 5 × 104 sporulated oocysts of Eimeria maxima. The live performance was evaluated on day 0 (day of inoculation), 5°, 10°, 15°, 25°, and 35° dpi, being slaughtered by cervical dislocation, fifteen birds/group. Although the sum in meat production was higher in the control group, the weight of the heart and gizzard of the experimental animals showed no significant difference, while the liver had difference on day 5°, 15°, and 35° dpi. The pathologic evaluation showed congested mucosa and presence of large amounts of mucus at 6 dpi. Therefore, it is concluded that the dose of 5 × 104  E. maxima inoculated in the experimental group was enough to cause harm to the animal organism. PMID:26464925

  11. Cryptosporidium surveillance and risk factors in the United States.

    Science.gov (United States)

    Yoder, Jonathan S; Beach, Michael J

    2010-01-01

    Surveillance for Cryptosporidium in the United States indicates that the reported incidence of infection has increased dramatically since 2004. The reasons for this increase are unclear but might be caused by an actual increase in incidence, improved surveillance, improved awareness about cryptosporidiosis, and/or increases in testing practices resulting from the licensing of the first-ever treatment for cryptosporidiosis. While regional differences remain, the incidence of cryptosporidiosis appears to be increasing across the United States. Onset of illness is most common during the summer, particularly among younger children. Cryptosporidiosis case reporting also influences outbreak detection and reporting; the recent rise in cases coincides with an increase in the number of reported cryptosporidiosis outbreaks, particularly in treated recreational water venues. Risk factors include ingesting contaminated recreational or drinking water, exposure to infected animals, having close contacts with cryptosporidiosis, travel to disease-endemic areas, and ingestion of contaminated food. Advances in molecular characterization of clinical specimens have improved our understanding of the changing epidemiology and risk factors. Prevention and control of cryptosporidiosis requires continued efforts to interrupt the transmission of Cryptosporidium through water, food, and contact with infected persons or animals. Of particular importance is continued improvement and monitoring of drinking water treatment and advances in the design, operation, and management of recreational water venues coupled with behavioral changes among the swimming public. Published by Elsevier Inc.

  12. Drug treatment and novel drug target against Cryptosporidium

    Directory of Open Access Journals (Sweden)

    Gargala G.

    2008-09-01

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

  13. Oleylphosphocholine (OlPC) arrests Cryptosporidium parvum growth in vitro and prevents lethal infection in interferon gamma receptor knock-out mice.

    Science.gov (United States)

    Sonzogni-Desautels, Karine; Renteria, Axel E; Camargo, Fabio V; Di Lenardo, Thomas Z; Mikhail, Alexandre; Arrowood, Michael J; Fortin, Anny; Ndao, Momar

    2015-01-01

    Cryptosporidium parvum is a species of protozoa that causes cryptosporidiosis, an intestinal disease affecting many mammals including humans. Typically, in healthy individuals, cryptosporidiosis is a self-limiting disease. However, C. parvum can cause a severe and persistent infection that can be life-threatening for immunocompromised individuals, such as AIDS patients. As there are no available treatments for these patients that can cure the disease, there is an urgent need to identify treatment options. We tested the anti-parasitic activity of the alkylphosphocholine oleylphosphocholine (OlPC), an analog of miltefosine, against C. parvum in in vitro and in vivo studies. In vitro experiments using C. parvum infected human ileocecal adenocarcinoma cells (HCT-8 cells) showed that OlPC has an EC50 of 18.84 nM. Moreover, no cell toxicity has been seen at concentrations ≤50 μM. C57BL/6 interferon gamma receptor knock-out mice, were infected by gavage with 4000 C. parvum oocysts on Day 0. Oral treatments, with OlPC, miltefosine, paromomycin or PBS, began on Day 3 post-infection for 10 days. Treatment with OlPC, at 40 mg/kg/day resulted in 100% survival, complete clearance of parasite in stools and a 99.9% parasite burden reduction in the intestines at Day 30. Doses of 30 and 20 mg/kg/day also demonstrated an increased survival rate and a dose-dependent parasite burden reduction. Mice treated with 10 mg/kg/day of miltefosine resulted in 50% survival at Day 30. In contrast, control mice, treated with PBS or 100 mg/kg/day of paromomycin, died or had to be euthanized between Days 6 and 13 due to severe illness. Results of parasite burden were obtained by qPCR and cross-validated by both flow cytometry of stool oocysts and histological sections of the ileum. Together, our results strongly support that OlPC represents a potential candidate for the treatment of C. parvum infections in immunocompromised patients.

  14. Oleylphosphocholine (OlPC arrests Cryptosporidium parvum growth in vitro and prevents lethal infection in interferon gamma receptor knock-out mice

    Directory of Open Access Journals (Sweden)

    Karine eSonzogni-Desautels

    2015-09-01

    Full Text Available Cryptosporidium parvum is a species of protozoa that causes cryptosporidiosis, an intestinal disease affecting many mammals including humans. Typically, in healthy individuals, cryptosporidiosis is a self-limiting disease. However, C. parvum can cause a severe and persistent infection that can be life-threatening for immunocompromised individuals, such as AIDS patients. As there are no available treatments for these patients that can cure the disease, there is an urgent need to identify treatment options. We tested the anti-parasitic activity of the alkylphosphocholine oleylphosphocholine (OlPC, an analog of miltefosine, against C. parvum in in vitro and in vivo studies. In vitro experiments using C. parvum infected human ileocecal adenocarcinoma cells (HCT-8 cells showed that OlPC has an EC50 of 18.84 nM. Moreover, no cell toxicity has been seen at concentrations ≤50 µM. C57BL/6 interferon gamma receptor knock-out mice, were infected by gavage with 4000 C. parvum oocysts on Day 0. Oral treatments, with OlPC, miltefosine, paromomycin or PBS, began on Day 3 post-infection for 10 days. Treatment with OlPC, at 40 mg/kg/day resulted in 100% survival, complete clearance of parasite in stools and a 99.9% parasite burden reduction in the intestines at Day 30. Doses of 30 mg/kg/day and 20 mg/kg/day also demonstrated an increased survival rate and a dose-dependent parasite burden reduction. Mice treated with 10 mg/kg/day of miltefosine resulted in 50% survival at Day 30. In contrast, control mice, treated with PBS or 100 mg/kg/day of paromomycin, died or had to be euthanized between Days 6 and 13 due to severe illness. Results of parasite burden were obtained by qPCR and cross-validated by both flow cytometry of stool oocysts and histological sections of the ileum. Together, our results strongly support that OlPC represents a potential candidate for the treatment of C. parvum infections in immunocompromised patients.

  15. Public health significance of zoonotic Cryptosporidium species in wildlife: Critical insights into better drinking water management

    Directory of Open Access Journals (Sweden)

    Alireza Zahedi

    2016-04-01

    Full Text Available Cryptosporidium is an enteric parasite that is transmitted via the faecal–oral route, water and food. Humans, wildlife and domestic livestock all potentially contribute Cryptosporidium to surface waters. Human encroachment into natural ecosystems has led to an increase in interactions between humans, domestic animals and wildlife populations. Increasing numbers of zoonotic diseases and spill over/back of zoonotic pathogens is a consequence of this anthropogenic disturbance. Drinking water catchments and water reservoir areas have been at the front line of this conflict as they can be easily contaminated by zoonotic waterborne pathogens. Therefore, the epidemiology of zoonotic species of Cryptosporidium in free-ranging and captive wildlife is of increasing importance. This review focuses on zoonotic Cryptosporidium species reported in global wildlife populations to date, and highlights their significance for public health and the water industry.

  16. Public health significance of zoonotic Cryptosporidium species in wildlife: Critical insights into better drinking water management.

    Science.gov (United States)

    Zahedi, Alireza; Paparini, Andrea; Jian, Fuchun; Robertson, Ian; Ryan, Una

    2016-04-01

    Cryptosporidium is an enteric parasite that is transmitted via the faecal-oral route, water and food. Humans, wildlife and domestic livestock all potentially contribute Cryptosporidium to surface waters. Human encroachment into natural ecosystems has led to an increase in interactions between humans, domestic animals and wildlife populations. Increasing numbers of zoonotic diseases and spill over/back of zoonotic pathogens is a consequence of this anthropogenic disturbance. Drinking water catchments and water reservoir areas have been at the front line of this conflict as they can be easily contaminated by zoonotic waterborne pathogens. Therefore, the epidemiology of zoonotic species of Cryptosporidium in free-ranging and captive wildlife is of increasing importance. This review focuses on zoonotic Cryptosporidium species reported in global wildlife populations to date, and highlights their significance for public health and the water industry.

  17. Cryptosporidium outbreak in calves on a large dairy farm

    DEFF Research Database (Denmark)

    Niine, Tarmo; Dorbek-Kolin, Elisabeth; Lassen, Brian

    2018-01-01

    of life. HP concentration and HL treatment were negatively associated with weight gain at 3 months of age. Cryptosporidium positive faecal samples were significantly (P Eimeria positive samples were not. Correct prophylactic treatment with HL delayed...

  18. Literature Reference for Cryptosporidium spp. (Applied and Environmental Microbiology. 2007. 73(13): 4218–4225)

    Science.gov (United States)

    Procedures are described for analysis of drinking water samples and may be adapted for assessment of solid, particulate, aerosol, and liquid samples. The method uses real-time PCR for identification of Cryptosporidium spp.

  19. Cryptosporidium in drinking water: Evaluation of the ILSI quantitative risk assessment framework

    NARCIS (Netherlands)

    Teunis PFM; Havelaar AH; IMA; MGB

    1999-01-01

    In dit rapport wordt kwantitatieve risicoschatting voor Cryptosporidium parvum in drinkwater beschreven, voor inwoners van een stad met kraanwater dat is bereid door middel van conventionele behandeling van oppervlaktewater. De nadruk ligt op het geven van een kwantitatieve beschrijving van

  20. SPECIES AND STRAIN-SPECIFIC TYPING OF CRYPTOSPORIDIUM PARASITES IN CLINICAL AND ENVIRONMENTAL SAMPLES

    Science.gov (United States)

    Cryptosporidiosis has recently attracted attention as an emerging water borne and food borne disease as well as an opportunistic infection in HIV infected indivduals. The lack of genetic information, however, has resulted in confusion in the taxonomy of Cryptosporidium parasites ...

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

    Science.gov (United States)

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

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

    Science.gov (United States)

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

  3. Acute diarrhoea associated with Cryptosporidium sp in Belém, Brazil (preliminary report)

    OpenAIRE

    Loureiro, Edvaldo Carlos Brito; Linhares, Alexandre da Costa; Mata, Leonardo

    1986-01-01

    Cryptosporidium sp was detected in faeces from three children suffering from acute diarrhoea. In two cases no other concomitant agents were detected and in a 3rd. this agent was associated with Entamoeba histolytic, Entamoeba coli, Endolimax nana, Chilomastix mesnili and Pentatricbomonas hominis. Amostras de Cryptosporidium sp foram detectadas das fezes de três crianças com diarréia aguda. Em dois casos nenhum outro agente foi registrado, concomitantemente, e no terceiro caso, esse coccidi...

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

    OpenAIRE

    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 ...

  5. Infectivity, pathogenicity, and genetic characteristics of mammalian gastric Cryptosporidium spp. in domestic ruminants

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Sak, Bohumil; Květoňová, Dana; Ditrich, Oleg; Hofmannová, L.; Modrý, David; Vítovec, J.; Xiao, L.

    2008-01-01

    Roč. 153, 3/4 (2008), s. 363-367 ISSN 0304-4017 R&D Projects: GA ČR GA524/05/0992; GA ČR GD524/03/H133 Institutional research plan: CEZ:AV0Z60220518 Keywords : Cryptosporidium andersoni * Cryptosporidium muris * ruminants Subject RIV: EG - Zoology Impact factor: 2.039, year: 2008

  6. A novel photoinduced electron transfer (PET) primer technique for rapid real-time PCR detection of Cryptosporidium spp

    International Nuclear Information System (INIS)

    Jothikumar, N.; Hill, Vincent R.

    2013-01-01

    Highlights: •Uses a single-labeled fluorescent primer for real-time PCR. •The detection sensitivity of PET PCR was comparable to TaqMan PCR. •Melt curve analysis can be performed to confirm target amplicon production. •Conventional PCR primers can be converted to PET PCR primers. -- Abstract: We report the development of a fluorescently labeled oligonucleotide primer that can be used to monitor real-time PCR. The primer has two parts, the 3′-end of the primer is complimentary to the target and a universal 17-mer stem loop at the 5′-end forms a hairpin structure. A fluorescent dye is attached to 5′-end of either the forward or reverse primer. The presence of guanosine residues at the first and second position of the 3′ dangling end effectively quenches the fluorescence due to the photo electron transfer (PET) mechanism. During the synthesis of nucleic acid, the hairpin structure is linearized and the fluorescence of the incorporated primer increases several-fold due to release of the fluorescently labeled tail and the absence of guanosine quenching. As amplicons are synthesized during nucleic acid amplification, the fluorescence increase in the reaction mixture can be measured with commercially available real-time PCR instruments. In addition, a melting procedure can be performed to denature the double-stranded amplicons, thereby generating fluorescence peaks that can differentiate primer dimers and other non-specific amplicons if formed during the reaction. We demonstrated the application of PET-PCR for the rapid detection and quantification of Cryptosporidium parvum DNA. Comparison with a previously published TaqMan® assay demonstrated that the two real-time PCR assays exhibited similar sensitivity for a dynamic range of detection of 6000–0.6 oocysts per reaction. PET PCR primers are simple to design and less-expensive than dual-labeled probe PCR methods, and should be of interest for use by laboratories operating in resource

  7. A novel photoinduced electron transfer (PET) primer technique for rapid real-time PCR detection of Cryptosporidium spp

    Energy Technology Data Exchange (ETDEWEB)

    Jothikumar, N., E-mail: jin2@cdc.gov; Hill, Vincent R.

    2013-06-28

    Highlights: •Uses a single-labeled fluorescent primer for real-time PCR. •The detection sensitivity of PET PCR was comparable to TaqMan PCR. •Melt curve analysis can be performed to confirm target amplicon production. •Conventional PCR primers can be converted to PET PCR primers. -- Abstract: We report the development of a fluorescently labeled oligonucleotide primer that can be used to monitor real-time PCR. The primer has two parts, the 3′-end of the primer is complimentary to the target and a universal 17-mer stem loop at the 5′-end forms a hairpin structure. A fluorescent dye is attached to 5′-end of either the forward or reverse primer. The presence of guanosine residues at the first and second position of the 3′ dangling end effectively quenches the fluorescence due to the photo electron transfer (PET) mechanism. During the synthesis of nucleic acid, the hairpin structure is linearized and the fluorescence of the incorporated primer increases several-fold due to release of the fluorescently labeled tail and the absence of guanosine quenching. As amplicons are synthesized during nucleic acid amplification, the fluorescence increase in the reaction mixture can be measured with commercially available real-time PCR instruments. In addition, a melting procedure can be performed to denature the double-stranded amplicons, thereby generating fluorescence peaks that can differentiate primer dimers and other non-specific amplicons if formed during the reaction. We demonstrated the application of PET-PCR for the rapid detection and quantification of Cryptosporidium parvum DNA. Comparison with a previously published TaqMan® assay demonstrated that the two real-time PCR assays exhibited similar sensitivity for a dynamic range of detection of 6000–0.6 oocysts per reaction. PET PCR primers are simple to design and less-expensive than dual-labeled probe PCR methods, and should be of interest for use by laboratories operating in resource

  8. Cryptosporidium Species are Frequently Present but Rarely Detected in Clinical Samples from Children with Diarrhea in a Developed Country

    DEFF Research Database (Denmark)

    Skovgaards, Daniel M; Hartmeyer, Gitte N; Skov, Marianne N

    2018-01-01

    Two studies were done on cryptosporidiosis in children. A retrospective survey showed that from 2005 to 2015 Cryptosporidium species was detected by microscopy of stool from 0.25% of children with diarrhea. In a subsequent prospective study PCR detected Cryptosporidium species in 4 (1,3%) of 304...... children. Cryptosporidium species is as frequent as other intestinal pathogens in childhood diarrhea. Testing is relevant....

  9. Prevalence and genetic characterization of Cryptosporidium in yaks in Qinghai Province of China.

    Directory of Open Access Journals (Sweden)

    Rongsheng Mi

    Full Text Available The objective of this study was to determine the prevalence, species and subtypes of Cryptosporidium infecting yaks in the Qinghai Province of Northwestern China. The prevalence of Cryptosporidium spp. was detected by microscopy and nested-PCR. A total of 586 fecal samples were collected from yaks in 6 counties, of which 142 (24.2% samples tested positive for Cryptosporidium. The small subunit (SSU rRNA gene of fifty-five samples were amplified and sequenced successfully and demonstrated that Cryptosporidium bovis (31/55, 56.4% was the most common species, followed by C. parvum (16/55, 29.1% and C. ryanae (5/55, 9.0%. Mixed infections of C. parvum and C. bovis (n = 2, C. ryanae and C. bovis (n = 1 were also detected. All three species were found in yaks ranging in age from 2 years. Cryptosporidium was most commonly detected in spring (28.4%, followed by summer (20.9%, then winter (17.5%. Cryptosporidium parvum positive samples were subtyped using the 60 kDa glycoprotein (gp60 gene. Subtypes IIaA15G2R1 (n = 8, IIaA16G2R1 (n = 2, IIaA14G1R1 (n = 1, IIaA14G2R1 (n = 1 and IIaA16G3R1 (n = 1 were detected. All of these subtypes are zoonotic, and may pose a potential threat to human health.

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

    Directory of Open Access Journals (Sweden)

    Xiao-Xuan Zhang

    2015-01-01

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

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

    Science.gov (United States)

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

    2015-01-01

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

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

    Science.gov (United States)

    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. K. Koompapong et al., published by EDP Sciences, 2014

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

    Directory of Open Access Journals (Sweden)

    Koompapong Khuanchai

    2014-01-01

    Full Text Available 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.

  14. Antibody responses measured by various serologic tests in pigs orally inoculated with low numbers of Toxoplasma gondii oocysts

    DEFF Research Database (Denmark)

    Dubey, J. P.; Andrews, C.D.; Lind, Peter

    1996-01-01

    ) infective oocysts, and 6 pigs served as uninoculated controls. Blood (serum) samples were obtained at 1- to 3-week intervals until euthanasia. At necropsy, the brain, heart, and tongue of pigs were bioassayed in mice and cats for isolation of T gondii. Modified agglutination test (MAT), using whole, fixed...

  15. Eliminación de partículas similares a Cryptosporidium ooquistes por filtración directa ascendente utilizando un coagulante natural: estudio a escala piloto

    Directory of Open Access Journals (Sweden)

    Ingrith Marcela Romero Méndez

    2016-12-01

    Full Text Available En la presente investigación fueron realizados experimentos de filtración de flujo ascendente en  línea, a escala piloto, para evaluar el desempeño del coagulante natural quitosano en la extracción de microesferas de poliestireno con dimensiones similares a Cryptosporidium oocysts, en el tratamiento de aguas con turbidez natural aproximada de 30 uT, inducida con caolinita. Los parámetros evaluados fueron turbidez, materia orgánica (absorbancia a 254 nm y concentración de microesferas. Inicialmente, se realizaron pruebas de jarras para la construcción del diagrama de coagulación y la definición de las condiciones para ser adoptadas en experimentos de filtración. Los experimentos de filtración de flujo ascendente en línea se efectuaron en condiciones de coagulación de dosis óptimas, super-óptimas y sub-óptimas. El diagrama de coagulación mostró una región de un rendimiento óptimo de quitosano, correspondiente a los valores de turbidez del efluente de 0,5 uT a 0.25 uT en dosis entre 0,9 e 1,5 mg/L y pH valores entre 5 y 6. En los experimentos de filtración en condiciones de coagulación óptima la turbiedez promedio del efluente fue 0.29 uT durante el funcionamiento normal del sistema. En la coagulación de esta condición la extracción de microesferas osciló entre 4,3 log e log 4.9. En condiciones de coagulación super-óptima y subóptima, se afectó el rendimiento del filtro con incrementos en la turbidez del efluente y fue acompañado por la reducción en la extracción de microesferas con respecto a la condición de coagulación óptima del orden de 0,65 a 0,98 log, respectivamente, durante el funcionamiento normal del sistema, lo que indica que estas condiciones de coagulación conducen a aumentar el número de partículas con dimensiones similares a Cryptosporidium oocysts en el efluente del filtro, lo cual puede comprometer la calidad microbiológica del agua producida. Finalmente, en todas las condiciones de

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

    DEFF Research Database (Denmark)

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

    2003-01-01

    and rectum. The unintended presence of rotavirus in some of the experimental animals revealed an additive or synergistic effect between rotavirus and C. parvum as indicated by prolonged diarrhoea, increased oocyst shedding, decreased weight gain and elevated levels of serum haptoglobin and serum amyloid...... A (SAA) in piglets infected simultaneously with both pathogens. The difference in daily weight gain between infected and control animals was significant only for piglets co-infected with rotavirus. The acute phase response of haptoglobin and SAA was characterised by a large individual variation....... In piglets, co-infected with rotavirus, the levels of serum haptoglobin were 3.5 and 4.6 times higher in the infected versus the controls 6 and 9 dpi, respectively (mean values: 2411 mug/ml +/- S.D. 2023 and 1840 mug/ml +/- S.D. 1697). In the controls infected with rotavirus, peak haptoglobin concentration...

  17. Eimeriosis in Danish Dairy Calves – Correlation between Species, Oocyst Excretion and Diarrhoea

    DEFF Research Database (Denmark)

    Enemark, Heidi L.; Dahl, Jan; Enemark, Jörg M Dehn

    2013-01-01

    The study collected up-to-date data on prevalence and importance of Eimeria infections in Danish dairy calves with suspected clinical eimeriosis and analysed correlation between Eimeria spp., oocyst excretion and diarrhoea. From October 2010 through August 2011, veterinarians collected faecal...... determined, along with opg values for the specific Eimeria spp. Association between opg and faeces consistency was evaluated in a multinomial, logistic regression model. Overall prevalence of Eimeria spp. was 96.2 % with a prevalence of 60.9 % in individual calves. E. zuernii and/or E. bovis were detected...... in 88.5 % of herds and 41.5 % of the calves. Mean opg was 2,040 (range 0–114,000) in the calves, of which 18.1 % had opg values ≥ 1,000. A total of 12 Eimeria spp. was found with the following calf prevalences: E. ellipsoidalis (37 %), E. zuernii (32 %), E. bovis (28 %), E. cylindrica (23 %), E...

  18. Comparative genomic analysis reveals occurrence of genetic recombination in virulent Cryptosporidium hominis subtypes and telomeric gene duplications in Cryptosporidium parvum.

    Science.gov (United States)

    Guo, Yaqiong; Tang, Kevin; Rowe, Lori A; Li, Na; Roellig, Dawn M; Knipe, Kristine; Frace, Michael; Yang, Chunfu; Feng, Yaoyu; Xiao, Lihua

    2015-04-18

    Cryptosporidium hominis is a dominant species for human cryptosporidiosis. Within the species, IbA10G2 is the most virulent subtype responsible for all C. hominis-associated outbreaks in Europe and Australia, and is a dominant outbreak subtype in the United States. In recent yearsIaA28R4 is becoming a major new subtype in the United States. In this study, we sequenced the genomes of two field specimens from each of the two subtypes and conducted a comparative genomic analysis of the obtained sequences with those from the only fully sequenced Cryptosporidium parvum genome. Altogether, 8.59-9.05 Mb of Cryptosporidium sequences in 45-767 assembled contigs were obtained from the four specimens, representing 94.36-99.47% coverage of the expected genome. These genomes had complete synteny in gene organization and 96.86-97.0% and 99.72-99.83% nucleotide sequence similarities to the published genomes of C. parvum and C. hominis, respectively. Several major insertions and deletions were seen between C. hominis and C. parvum genomes, involving mostly members of multicopy gene families near telomeres. The four C. hominis genomes were highly similar to each other and divergent from the reference IaA25R3 genome in some highly polymorphic regions. Major sequence differences among the four specimens sequenced in this study were in the 5' and 3' ends of chromosome 6 and the gp60 region, largely the result of genetic recombination. The sequence similarity among specimens of the two dominant outbreak subtypes and genetic recombination in chromosome 6, especially around the putative virulence determinant gp60 region, suggest that genetic recombination plays a potential role in the emergence of hyper-transmissible C. hominis subtypes. The high sequence conservation between C. parvum and C. hominis genomes and significant differences in copy numbers of MEDLE family secreted proteins and insulinase-like proteases indicate that telomeric gene duplications could potentially contribute to

  19. Prevalence of Giardia sp. Cryptosporidium parvum and Cryptosporidium andersoni (syn. C. muris) [correction of Cryptosporidium parvum and Cryptosporidium muris (C. andersoni)] in 109 dairy herds in five counties of southeastern New York.

    Science.gov (United States)

    Wade, S E; Mohammed, H O; Schaaf, S L

    2000-11-01

    A cross-sectional study was undertaken to determine the prevalence of Giardia sp. (G. duodenalis group), Cryptosporidium parvum and Cryptosporidium andersoni (C. muris) [corrected] in dairy cattle in three different age groups, and to evaluate the association of age and season with prevalence. One hundred and nine dairy farms, from a total of 212 farms, in five counties of southeastern New York volunteered to participate. On these farms, 2943 fecal samples were collected from three defined age groups. The farms were randomly assigned for sampling within the four seasons of the year. Each farm was visited once during the study period from March 1993 to June 1994 to collect fecal samples. Demographic data on the study population was collected at the time of sampling by interviewing the farm owner or manager. At collection, fecal samples were scored as diarrheic or non-diarrheic, and each condition was later related to positive or negative infection with these parasites. Fecal samples were processed using a quantitative centrifugation concentration flotation technique and enumerated using bright field and phase contrast microscopy. In this study, the overall population prevalence for Giardia sp. was 8.9%; C. parvum, 0.9%; and C. muris, 1.1%. When considering animals most at the risk of infection (those younger than 6 months of age) Giardia sp. and C. parvum was found in 20.1 and 2.4% of the animals, respectively. Giardia sp. and C. muris were found in all age groups. There was no significant seasonal pattern of infection for any of these parasites.

  20. Modelling the impact of sanitation, population growth and urbanization on human emissions of Cryptosporidium to surface waters—a case study for Bangladesh and India

    NARCIS (Netherlands)

    Vermeulen, L.C.; Kraker, Dummy; Hofstra, N.; Kroeze, C.; Medema, G.J.

    2015-01-01

    Cryptosporidium is a protozoan parasite that can cause diarrhoea. Human faeces are an important source of Cryptosporidium in surface waters. We present a model to study the impact of sanitation, urbanization and population growth on human emissions of Cryptosporidium to surface waters. We build on a

  1. Modelling the impact of sanitation, population growth and urbanization on human emissions of cryptosporidium to surface waters : A case study for Bangladesh and India

    NARCIS (Netherlands)

    Vermeulen, L.C.; Kraker, J.; Hofstra, N.; Kroeze, C.; Medema, G.

    2015-01-01

    Cryptosporidium is a protozoan parasite that can cause diarrhoea. Human faeces are an important source of Cryptosporidium in surface waters. We present a model to study the impact of sanitation, urbanization and population growth on human emissions of Cryptosporidium to surface waters. We build on a

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

    Science.gov (United States)

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

    2011-08-01

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

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

    Directory of Open Access Journals (Sweden)

    Daniel J Becker

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

  4. Monoclonal Antibodies to Intracellular Stages of Cryptosporidium parvum Define Life Cycle Progression In Vitro.

    Science.gov (United States)

    Wilke, Georgia; Ravindran, Soumya; Funkhouser-Jones, Lisa; Barks, Jennifer; Wang, Qiuling; VanDussen, Kelli L; Stappenbeck, Thaddeus S; Kuhlenschmidt, Theresa B; Kuhlenschmidt, Mark S; Sibley, L David

    2018-06-27

    Among the obstacles hindering Cryptosporidium research is the lack of an in vitro culture system that supports complete life development and propagation. This major barrier has led to a shortage of widely available anti- Cryptosporidium antibodies and a lack of markers for staging developmental progression. Previously developed antibodies against Cryptosporidium were raised against extracellular stages or recombinant proteins, leading to antibodies with limited reactivity across the parasite life cycle. Here we sought to create antibodies that recognize novel epitopes that could be used to define intracellular development. We identified a mouse epithelial cell line that supported C. parvum growth, enabling immunization of mice with infected cells to create a bank of monoclonal antibodies (MAbs) against intracellular parasite stages while avoiding the development of host-specific antibodies. From this bank, we identified 12 antibodies with a range of reactivities across the parasite life cycle. Importantly, we identified specific MAbs that can distinguish different life cycle stages, such as trophozoites, merozoites, type I versus II meronts, and macrogamonts. These MAbs provide valuable tools for the Cryptosporidium research community and will facilitate future investigation into parasite biology. IMPORTANCE Cryptosporidium is a protozoan parasite that causes gastrointestinal disease in humans and animals. Currently, there is a limited array of antibodies available against the parasite, which hinders imaging studies and makes it difficult to visualize the parasite life cycle in different culture systems. In order to alleviate this reagent gap, we created a library of novel antibodies against the intracellular life cycle stages of Cryptosporidium We identified antibodies that recognize specific life cycle stages in distinctive ways, enabling unambiguous description of the parasite life cycle. These MAbs will aid future investigation into Cryptosporidium biology and

  5. SYNERGY IN SEQUENTIAL INACTIVATION OF CRYPTOSPORIDIUM PARVUM WITH OZONE/FREE CHLORINE AND OZONE/MONOCHLORAMINE. (R826830)

    Science.gov (United States)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  6. Prevalence and age-related infection of Cryptosporidium suis, C. muris and Cryptosporidium pig genotype II in pigs on a farm complex in the Czech Republic

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Hanzlíková, D.; Sak, Bohumil; Květoňová, Dana

    2009-01-01

    Roč. 160, 3/4 (2009), s. 319-322 ISSN 0304-4017 R&D Projects: GA ČR GP523/07/P117 Institutional research plan: CEZ:AV0Z60220518 Keywords : Cryptosporidium infection * age specificity * pigs Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.278, year: 2009

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

    Czech Academy of Sciences Publication Activity Database

    Laatamna, A.E.; Wagnerová, Pavla; Sak, Bohumil; Květoňová, Dana; Xiao, L.; Rost, M.; McEvoy, J.; Saadi, A.R.; Aissi, M.; Kváč, Martin

    2015-01-01

    Roč. 208, 3-4 (2015), s. 135-142 ISSN 0304-4017 R&D Projects: GA ČR GA15-01090S Institutional support: RVO:60077344 Keywords : Horses * Donkeys * Cryptosporidium spp. * Encephalitozoon spp. * Enterocytozoon bieneusi * Molecular prevalence Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.242, year: 2015

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

    Science.gov (United States)

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

    2016-02-01

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

  9. Cryptosporidium homai n. sp. (Apicomplexa: Cryptosporidiiae) from the guinea pig (Cavia porcellus).

    Science.gov (United States)

    Zahedi, Alireza; Durmic, Zoey; Gofton, Alexander W; Kueh, Susan; Austen, Jill; Lawson, Malcolm; Callahan, Lauren; Jardine, John; Ryan, Una

    2017-10-15

    The morphological, biological, and molecular characterisation of a new Cryptosporidium species from the guinea pig (Cavia porcellus) are described, and the species name Cryptosporidium homai n. sp. is proposed. Histological analysis conducted on a post-mortem sample from a guinea pig euthanised due to respiratory distress, identified developmental stages of C. homai n. sp. (trophozoites and meronts) along the intestinal epithelium. Molecular analysis at 18S rRNA (18S), actin and hsp70 loci was then conducted on faeces from an additional 7 guinea pigs positive for C. homai n. sp. At the 18S, actin and hsp70 loci, C. homai n. sp. exhibited genetic distances ranging from 3.1% to 14.3%, 14.4% to 24.5%, and 6.6% to 20.9% from other Cryptosporidium spp., respectively. At the 18S locus, C. homai n. sp. shared 99.1% similarity with a previously described Cryptosporidium genotype in guinea pigs from Brazil and it is likely that they are the same species, however this cannot be confirmed as actin and hsp70 sequences from the Brazilian guinea pig genotype are not available. Phylogenetic analysis of concatenated 18S, actin and hsp70 sequences showed that C. homai n. sp. exhibited 9.1% to 17.3% genetic distance from all other Cryptosporidium spp. This clearly supports the validity of C. homai n. sp. as a separate species. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Lipid and glucose metabolism of broilers (Gallus gallus domesticus experimentally infected with Eimeria acervulina Tyzzer, 1929 oocysts

    Directory of Open Access Journals (Sweden)

    FLC Freitas

    2008-09-01

    Full Text Available Lipid and glucose metabolism of 76 ten-day-old Cobb male broilers, experimentally infected with Eimeria acervulina, was studied for 30 days. Birds were distributed in 2 groups: one infected with 1x10(6 E. acervulina sporulated oocysts, and the other inoculated with distilled water. Pathological e biochemical liver changes were assessed, as well as plasma glucose concentrations and total cholesterol, HDL, LDL, VLDL, fatty-acid, and triglyceride levels in the serum. The infected broilers presented hypoglycemia associated with a reduction in liver glycogen. In addition, these birds developed fatty liver, and there were changes in all lipid classes in the serum. Lipid and glucose metabolism was dramatically changed in broilers experimentally infected with 1x10(6 E. acervulina oocysts.

  11. Development of resistance to coccidiosis in the absence of merogonic development using X-irradiated Eimeria acervulina oocysts

    International Nuclear Information System (INIS)

    Jenkins, M.C.; Augustine, P.C.; Barta, J.R.; Castle, M.D.; Danforth, H.D.

    1991-01-01

    Sporulated oocysts of the protozoan Eimeria acervulina were subjected to 0, 10, 15, 20, or 30 krad of X-irradiation and inoculated into susceptible outbred chickens to determine if radioattenuated coccidia could induce protection against parasite challenge. Irradiation treatment had an appreciable dose-dependent effect on parasite development. Insignificant numbers of oocysts were produced by chickens inoculated with parasites that had been exposed to greater than 10 krad X-irradiation. Sporozoites exposed to 15 or 20 krad irradiation conferred significant protection against the appearance of intestinal lesions after parasite challenge. Sporozoites subjected to the highest dose level (30 krad) did not produce any significant level of protection. To investigate this phenomenon further and assess intracellular parasite development, susceptible outbred strains of chickens were administered either nonirradiated (0 krad) oocysts or oocysts that were exposed to an optimal dose (15 krad) or a high dose (30 krad) of X-irradiation. Immunofluorescence staining of tissue sections from each treatment group at various intervals after the initial administration of irradiated parasites indicated that sporozoites exposed to 15 krad irradiation were as capable of invading the host intestinal epithelium as nonirradiated sporozoites. However, at 48, 60, 72, and 96 hr, there was a marked reduction in merogonic development in groups receiving irradiated sporozoites compared to those inoculated with nonirradiated parasites. The latter parasites underwent profuse merogonic development; in contrast, irradiated parasites demonstrated little (15 krad) or no (30 krad) merogonic development. These results suggest that induction of a protective immune response occurs during a critical period early in intracellular development of E. acervulina

  12. Comparative Response of the Nigerian Indigenous and Broiler Chickens to a Field Caecal Isolate of Eimeria Oocysts

    Directory of Open Access Journals (Sweden)

    Lucas Atehmengo Ngongeh

    2017-01-01

    Full Text Available Response of Nigerian indigenous (local and broiler chickens to experimental Eimeria infections was investigated by measures of clinical signs, packed cell volume (PCV, body weights (BW, feed consumption, faecal oocyst counts (oocyst per gram, and microscopic intestinal lesions. Three-week-old chickens of each breed received single pulse infections with 2500, 5000, and 100.000 sporulated Eimeria oocysts. Infected birds were dull and passed bloody diarrhoea. OPG showed a dose related response but no significant difference between groups (P>0.05. OPG was significantly higher in local chickens (P<0.05 and varied significantly with time (P<0.05. PCV declined significantly in infected birds within breeds and groups (P<0.05; however, the decline in PCV was significantly greater in broilers (P<0.05. Both breeds had significant BW gains (P<0.05. BW gain varied between groups being significantly higher in the uninfected control broilers than in the infected broilers (P<0.05. Comparatively, broilers gained significantly more BW than their local counterparts (P<0.05. Feed intake increased significantly with time (P<0.05 in both breeds. The Eimeria isolate was pathogenic to both breeds of chicken although clinical signs and lesions were more severe in indigenous chickens suggesting the breed’s more susceptibility.

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

    Science.gov (United States)

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

    2009-03-01

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

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

    DEFF Research Database (Denmark)

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

    2001-01-01

    In community-based studies conducted from 1991 to 1997 in Guinea-Bissau, West Africa, stool specimens from children aged less than 5 years with diarrhoea were routinely examined for enteric parasites. Cryptosporidium parvum, found in 7.7% of 4,922 samples, was the second most common parasite......, exceeded only by Giardia lamblia which was found in 14.8% of the samples. The highest prevalence of cryptosporidium was found in children aged 6-11 months, whereas the prevalence of other enteric parasites increased with age. Cryptosporidiosis showed a marked seasonal variation, with peak prevalences found...... consistently at the beginning of or just before the rainy seasons, May through July. By contrast, no seasonality was found for the enteric parasites Giardia lamblia or Entamoeba histolytica. We conclude that Cryptosporidium parvum is an important pathogen in children with diarrhoea....

  15. Proliferative enteritis in leopard geckos (Eublepharis macularius) associated with Cryptosporidium sp. infection.

    Science.gov (United States)

    Terrell, Scott P; Uhl, Elizabeth W; Funk, Richard S

    2003-03-01

    Twenty-three leopard geckos (Eublepharis macularius) with various clinical histories of weight loss, anorexia, lethargy, and diarrhea were submitted either intact or as biopsy specimens to the University of Florida Anatomic Pathology Service. Gross necropsy findings in the intact geckos included marked reduction of subcutaneous adipose tissue stores at the tail base and mild thickening and reddening of the small intestine. Histologic examination revealed Cryptosporidium sp. infection associated with hyperplasia and mononuclear inflammation of the small intestine in all geckos. Parasites and lesions were only rarely observed in the stomach and large intestine of geckos. The histologic and ultrastructural lesions in the small intestine of leopard geckos infected with Cryptosporidium sp. have not been well characterized previously. This report implicates Cryptosporidium sp. as the cause of disease in the geckos and describes the range of histologic lesions observed.

  16. Cryptosporidium, Giardia and Enterocytozoon bieneusi in cats from Bogota (Colombia) and genotyping of isolates.

    Science.gov (United States)

    Santín, Mónica; Trout, James M; Vecino, Jesús A Cortés; Dubey, J P; Fayer, Ronald

    2006-11-05

    The prevalence of Cryptosporidium, Giardia, and Enterocytozoon bieneusi in cats from Bogota (Colombia) was determined from fecal specimens and scrapings of duodenal and ileal mucosa screened by PCR. All PCR-positive specimens were sequenced to determine the genotype(s) present. Of 46 cats, 6 (13%) were positive for Cryptosporidium, 5 (11%) were infected with C. felis and one (2%) with C. muris. Three (6.5%) cats were infected with Giardia duodenalis Assemblage F. Eight (17%) cats were infected with four genotypes of E. bieneusi: genotype D-like (9%), K (4%), Peru 10 (2%), and Peru 5 (2%). This is the first report on the presence of zoonotic species/genotypes of Cryptosporidium and E. bieneusi in cats in Colombia.

  17. Cryptosporidiosis outbreak in a child day-care center caused by an unusual Cryptosporidium hominis subtype.

    Science.gov (United States)

    Goñi, Pilar; Almagro-Nievas, Diego; Cieloszyk, Joanna; Lóbez, Silvia; Navarro-Marí, José María; Gutiérrez-Fernández, José

    2015-12-01

    This work describes the genetic characterization of Cryptosporidium and Giardia involved in an outbreak in a nursery school in Granada, Spain, that affected seven children under the age of 4. Nucleic acids were extracted from the seven stool samples positive to Cryptosporidium or Giardia by microscopy and/or immunochromatography. The species and subtypes of Cryptosporidium were identified by PCR-RFLP and PCR of the SSUrRNA and gp60 genes, respectively. The assemblages of Giardia duodenalis isolates were characterized by PCR of the tpi gene. PCR products were sequenced and analyzed. All of the isolates were positive for Cryptosporidium hominis. Five of them belonged to subtype IaA11R2, one to subtype IbA10G2R2, and the other could not be identified. Three of these samples were positive for G. duodenalis by PCR, two belonging to the assemblage A, and the other one to assemblage B. This is the first report of Cryptosporidium hominis subtype IaA11R2 as a cause of an outbreak in Europe where subtype IbA10G2R2 is the most frequently identified. In the case of Giardia, an outbreak could not be confirmed because of the low number of positive samples and the low genetic variability of the amplified fragments for assemblage A of tpi gene. A new subtype, of Cryptosporidium hominis named IaA11R2, has been described as a cause of an outbreak in a nursery school in Granada, Spain. However an outbreak of giardiasis could not be confirmed. Copyright © 2015 Elsevier España, S.L.U. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  18. High association of Cryptosporidium spp. infection with colon adenocarcinoma in Lebanese patients.

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    Marwan Osman

    Full Text Available The association between Cryptosporidium and human colon cancer has been reported in different populations. However, this association has not been well studied. In order to add new strong arguments for a probable link between cryptosporidiosis and colon human cancer, the aim of this study was to determine prevalence and to identify species of Cryptosporidium among Lebanese patients.Overall, 218 digestive biopsies were collected in Tripoli, Lebanon, from three groups of patients: (i patients with recently diagnosed colon intraepithelial neoplasia/adenocarcinoma before any treatment (n = 72; (ii patients with recently diagnosed stomach intraepithelial neoplasia/adenocarcinoma before any treatment (n = 21; and (iii patients without digestive intraepithelial neoplasia/adenocarcinoma but with persistent digestive symptoms (n = 125. DNA extraction was performed from paraffin-embedded tissue. The presence of the parasite in tissues was confirmed by PCR, microscopic observation and immunofluorescence analysis. We identified a high rate (21% of Cryptosporidium presence in biopsies from Lebanese patients with recently diagnosed colonic neoplasia/adenocarcinoma before any treatment. This prevalence was significantly higher compared to 7% of Cryptosporidium prevalence among patients without colon neoplasia but with persistent gastrointestinal symptoms (OR: 4, CI: 1.65-9.6, P = 0.001. When the comparison was done against normal biopsies, the risk of infection increased 11-fold in the group of patients with colon adenocarcinoma (OR: 11.315, CI: 1.44-89.02, P = 0.003.This is the first study performed in Lebanon reporting the prevalence of Cryptosporidium among patients with digestive cancer. These results show that Cryptosporidium is strongly associated with human colon cancer being maybe a potential etiological agent of this disease.

  19. The Prevalence of Cryptosporidium spp. in Children, Taiz District, Yemen

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    AH Al-Shamiri

    2010-06-01

    Full Text Available Background: This is the first work done on cryptosporidiosis among the children in Taiz, Yemen.Methods: A number of 712 samples were collected from children of different ages (ranging from 1 month to 12 years from Dec 2006 to Aug 2007. The collected samples were examined by Sheather's sugar floatation and Modified Ziehl- Neelsen stain as well as ELISA methods. The test results were statistically analyzed by SPSS software.Results: The overall positive percentage was 43.7%. The higher incidence (36.2 % was oc­curred in males while the lowest incidence (32.7 % was observed in females (r= 0.876; P= 0.001. The correlation between infected cases and the type of drinking water was r =0.121. Among the cases examined by ELISA (92 cases, 26.1 % were infected. The correlation be­tween seropositivity and gender was r= 0.652 (P=0.031.Conclusion: Cryptosporidium spp. is a significant pathogen among children at Taiz. Fresh water supplies, education, eating habits and domestic animals are considered the main sources for transmission of cryptosporidiosis.

  20. Metabolic alterations in broiler chickens experimentally infected with sporulated oocysts of Eimeria maxima.

    Science.gov (United States)

    Freitas, Fagner Luiz da Costa

    2014-01-01

    Metabolic and morphometric alterations of the duodenal villi caused by parasitism of chickens by Eimeria maxima were evaluated, using 100 male Cobb birds, randomly distributed into two groups (control and infected). The infected group was inoculated with 0.5 ml of a solution containing 5 × 10³ sporulated oocysts of Eimeria maxima. Ten birds per sample were sacrificed on the 6th, 11th, 22nd and 41st days post-infection (dpi). In order to evaluate the alterations, samples of duodenum, jejunum and ileum fragments were collected after necropsy for histological analysis. Villus biometry was determined by means of a slide graduated in microns that was attached to a binocular microscope. To evaluate the biochemical data, 5 ml of blood were sampled from the birds before sacrifice. The statistical analyses were performed using the GraphPad 5 statistical software for Windows. Tukey's multiple comparison test (p maxima causes both qualitative and quantitative alterations to the structure of the intestinal villi, thereby interfering with the absorption of nutrients such as calcium, phosphorus, magnesium, protein and lipids, with consequent reductions in the birds' weights.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2010-03-29

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

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

    Science.gov (United States)

    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. Copyright © 2015 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Moyad Avazpoor

    2015-01-01

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

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

    Science.gov (United States)

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

    2016-11-01

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

  5. Description of the Oocysts of Three New Species of Eimeria (Apicomplexa: Eimeriidae from Iguanid Lizards (Sauria: Iguanidae of Central and South America

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    Daszak P

    1998-01-01

    Full Text Available Three new species of Eimeria are described from iguanid lizards of Central and South America. The oocysts of each species have no micropyles or residua and the sporocysts lack Stieda bodies, but all have a sporocyst residuum. Eimeria sanctaluciae n.sp. was found in the St. Lucia tree lizard, Anolis luciae, collected from the Maria Islands, Lesser Antilles. The oocysts are spherical to subspherical, averaging 17.3 x 16.5 µm, with a single layered colourless wall; about 60% contain polar granules. The sporocysts are ellipsoidal and average 7.7 x 5.5 µm. Eimeria liolaemi n.sp. was recovered from the blue-gold swift, Liolaemus taenius, from Chile. The oocysts are spherical to subspherical, measuring 21 x 20.1 µm with a single-layered colourless wall. The sporocysts are subspherical and average 7.4 x 6.8 µm. Eimeria caesicia n.sp. is described from the Brazilian collared iguanid, Tropidurus torquatus. The oocysts measure 27.4 x 23.7 µm, are spherical to subspherical, with a bilayered wall, the outer surface of which appears pale blue in colour, the thin, inner wall appearing brown, when viewed by direct light under the optical microscope. The sporocysts are subspherical and average 9.4 x 7.2 µm. Unnamed polysporocystid oocysts with dizoic sporocysts are reported from the faeces of the lesser St. Vincent tree lizard, Anolis trinitatis and the possibility of spurious parasitism briefly discussed. In addition, oocysts of an unnamed Isospora sp. with a smooth oocyst wall which closely resembles I. reui were recovered from A. trinitatis.

  6. Life cycle of Cryptosporidium muris in two rodents with different responses to parasitization

    Czech Academy of Sciences Publication Activity Database

    Melicharová, J.; Ilgová, J.; Kváč, Martin; Sak, Bohumil; Koudela, B.; Valigurová, A.

    2014-01-01

    Roč. 141, č. 2 (2014), s. 287-303 ISSN 0031-1820 R&D Projects: GA ČR(CZ) GAP505/11/1163; GA MŠk(CZ) LH11061 Institutional support: RVO:60077344 Keywords : cryptosporidia * development * gastric * oocyst * pathology * Type II merogony Subject RIV: EE - Microbiology, Virology Impact factor: 2.560, year: 2014

  7. The first report of Cryptosporidium spp. in Microtus fuscus (Qinghai vole) and Ochotona curzoniae (wild plateau pika) in the Qinghai-Tibetan Plateau area, China.

    Science.gov (United States)

    Zhang, Xueyong; Jian, Yingna; Li, Xiuping; Ma, Liqing; Karanis, Gabriele; Karanis, Panagiotis

    2018-05-01

    Cryptosporidium is one of the most important genera of intestinal zoonotic pathogens, which can infect various hosts and cause diarrhoea. There is little available information about the molecular characterisation and epidemiological prevalence of Cryptosporidium spp. in Microtus fuscus (Qinghai vole) and Ochotona curzoniae (wild plateau pika) in the Qinghai-Tibetan Plateau area of Qinghai Province, Northwest China. Therefore, the aim of this study was to determine Cryptosporidium species/genotypes and epidemiological prevalence in these mammals by detecting the SSU rRNA gene by PCR amplification. The Cryptosporidium spp. infection rate was 8.9% (8/90) in Qinghai voles and 6.25% (4/64) in wild plateau pikas. Positive samples were successfully sequenced, and the following Cryptosporidium species were found: C. parvum, C. ubiquitum, C. canis and a novel genotype in Qinghai voles and C. parvum and a novel genotype in wild plateau pikas. This is the first report of Cryptosporidium infections in M. fuscus and wild O. curzoniae in Northwest China. The results suggest the possibility of Cryptosporidium species transmission among these two hosts, the environment, other animals and humans and provide useful molecular epidemiological data for the prevention and control of Cryptosporidium infections in wild animals and the surrounding environments. The results of the present study indicate the existence of Cryptosporidium species infections that have potential public health significance. This is the first report of Cryptosporidium multi-species infections in these animal hosts.

  8. Acute diarrhoea associated with Cryptosporidium sp in Belém, Brazil (preliminary report

    Directory of Open Access Journals (Sweden)

    Edvaldo Carlos Brito Loureiro

    1986-04-01

    Full Text Available Cryptosporidium sp was detected in faeces from three children suffering from acute diarrhoea. In two cases no other concomitant agents were detected and in a 3rd. this agent was associated with Entamoeba histolytic, Entamoeba coli, Endolimax nana, Chilomastix mesnili and Pentatricbomonas hominis.

  9. BIALLELIC POLYMORPHISM IN THE INTRON REGION OF B-TUBULIN GENE OF CRYPTOSPORIDIUM PARASITES

    Science.gov (United States)

    Nucleotide sequencing of polymerase chain reaction-amplified intron region of the Cryptosporidium parvum B-tubulin gene in 26 human and 15 animal isolates revealed distinct genetic polymorphism between the human and bovine genotypes. The separation of 2 genotypes of C. parvum is...

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

    Czech Academy of Sciences Publication Activity Database

    Stenger, B.L.S.; Clark, M.E.; Kváč, Martin; Khan, E.; Giddings, C.W.; Prediger, Jitka; McEvoy, J.M.

    2015-01-01

    Roč. 36, 2015-Dec (2015), s. 287-293 ISSN 1567-1348 R&D Projects: GA ČR GA15-01090S Institutional support: RVO:60077344 Keywords : Cryptosporidium * Tree squirrels * Ground squirrels * Host specificity * Zoonotic Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.591, year: 2015

  11. Novel Cryptosporidium bat genotypes III and IV in bats/nfrom the USA and Czech Republic

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Hořická, A.; Sak, Bohumil; Prediger, Jitka; Salát, J.; Širmarová, J.; Bartonička, T.; Clark, M.; Chelladurai, J.R.J.J.; Gillam, E.; McEvoy, J.

    2015-01-01

    Roč. 114, č. 10 (2015), s. 3917-3921 ISSN 0932-0113 R&D Projects: GA ČR GA15-01090S Institutional support: RVO:60077344 Keywords : Cryptosporidium * Bats * SSU * Actin * PCR Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.027, year: 2015

  12. Cryptosporidium parvum and Enterocytozoon bieneusi in American Mustangs and Chincoteague ponies

    Czech Academy of Sciences Publication Activity Database

    Wagnerová, Pavla; Sak, Bohumil; McEvoy, J.; Rost, M.; Sherwood, D.; Holcomb, K.; Kváč, Martin

    2016-01-01

    Roč. 162, MAR (2016), s. 24-27 ISSN 0014-4894 R&D Projects: GA ČR GA15-01090S Institutional support: RVO:60077344 Keywords : feral horses * Cryptosporidium * SSU * gp60 * Microsporidia * ITS Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 1.724, year: 2016

  13. An ultrastructural comparison of the attachment sites between Gregarina steini and Cryptosporidium muris

    Czech Academy of Sciences Publication Activity Database

    Valigurová, A.; Hofmannová, L.; Koudela, Břetislav; Vávra, Jiří

    2007-01-01

    Roč. 54, č. 6 (2007), s. 495-510 ISSN 1066-5234 R&D Projects: GA ČR GD524/03/H133 Institutional research plan: CEZ:AV0Z60220518 Keywords : Apicomplexa * gregarine * Cryptosporidium * feeder organelle * epimerite * parasites * ultrastructure Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.525, year: 2007

  14. Molecular characterization of Cryptosporidium isolates from pigs at slaughterhouses in South Bohemia, Czech Republic

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Sak, Bohumil; Hanzlíková, D.; Kotilová, J.; Květoňová, Dana

    2009-01-01

    Roč. 104, č. 2 (2009), s. 425-428 ISSN 0932-0113 R&D Projects: GA ČR GP523/07/P117 Institutional research plan: CEZ:AV0Z60220518 Keywords : Cryptosporidium spp. * slaughterhouse * pigs Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 1.721, year: 2009

  15. Prevalence and pathogenicity of Cryptosporidium andersoni in one herd of beef cattle

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Vítovec, J.

    2003-01-01

    Roč. 50, č. 9 (2003), s. 451-457 ISSN 0931-1793 Institutional research plan: CEZ:AV0Z6022909; CEZ:MSM 122200002 Keywords : cryptosporidiosis * cattle * Cryptosporidium andersoni Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 0.656, year: 2003

  16. Activated CD8+T cells contribute to clearance of gastric Cryptosporidium muris infections

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Kodádková, Alena; Sak, Bohumil; Květoňová, Dana; Jalovecká, M.; Rost, M.; Salát, Jiří

    2011-01-01

    Roč. 33, č. 4 (2011), 210-216 ISSN 0141-9838 R&D Projects: GA AV ČR KJB500960701 Institutional research plan: CEZ:AV0Z60220518 Keywords : CD4+T-lymphocytes * CD8+T-lymphocytes * Cryptosporidium muris * T-cell-mediated immunity Subject RIV: EC - Immunology Impact factor: 2.601, year: 2011

  17. Prevalence and molecular characterization of Cryptosporidium spp. in dairy cattle in South Bohemia, the Czech Republic

    Czech Academy of Sciences Publication Activity Database

    Ondráčková, Z.; Kváč, Martin; Sak, Bohumil; Květoňová, Dana; Rost, M.

    2009-01-01

    Roč. 165, 1/2 (2009), s. 141-144 ISSN 0304-4017 R&D Projects: GA ČR GP523/07/P117 Institutional research plan: CEZ:AV0Z60220518 Keywords : Cryptosporidium spp. * cattle * slaughterhouses Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.278, year: 2009

  18. Het rendement van de detectiemethode voor Cryptosporidium en Giardia in water

    NARCIS (Netherlands)

    Schets FM; Medema GJ; Schijven JF; MGB

    2004-01-01

    Nederlandse waterleidingbedrijven zijn verplicht om te berekenen of als gevolg van consumptie van drinkwater infectie met Cryptosporidium of Giardia kan optreden. De kans hierop moet kleiner dan een infectie per 10000 personen per jaar zijn. De berekening (risicoanalyse) wordt gebaseerd op de

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

    NARCIS (Netherlands)

    Ratsak CH; MGB

    1996-01-01

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

  20. Prevalence and pathogenicity of Cryptosporidium suis in pre- and post-weaned pigs

    Czech Academy of Sciences Publication Activity Database

    Vítovec, J.; Hamadejová, K.; Landová, L.; Kváč, Martin; Květoňová, Dana; Sak, Bohumil

    2006-01-01

    Roč. 53, č. 5 (2006), s. 239-243 ISSN 0931-1793 R&D Projects: GA ČR GA524/05/0992 Institutional research plan: CEZ:AV0Z60220518 Keywords : Cryptosporidium suis * piglets * pathogenicity Subject RIV: EG - Zoology Impact factor: 1.356, year: 2006

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

    Directory of Open Access Journals (Sweden)

    Fábio Tadeu Rodrigues REINA

    2016-01-01

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

  2. Human Primary Intestinal Epithelial Cells as an Improved In Vitro Model for Cryptosporidium parvum Infection

    Science.gov (United States)

    Cabada, Miguel M.; Nichols, Joan; Gomez, Guillermo; White, A. Clinton

    2013-01-01

    The study of human intestinal pathogens has been limited by the lack of methods for the long-term culture of primary human intestinal epithelial cells (PECs). The development of infection models with PECs would allow a better understanding of host-parasite interactions. The objective of this study was to develop a novel method for prolonged in vitro cultivation of PECs that can be used to study Cryptosporidium infection. We isolated intact crypts from human intestines removed during weight loss surgery. The fragments of intestinal layers were cultivated with culture medium supplemented with growth factors and antiapoptotic molecules. After 7 days, the PECs formed self-regenerating cell clusters, forming villi that resemble intestinal epithelium. The PECs proliferated and remained viable for at least 60 days. The cells expressed markers for intestinal stem cells, epithelial cells, and mature enterocytes. The PECs were infected with Cryptosporidium. In contrast to older models in which parasite numbers decay, the burden of parasites increased for >120 h. In summary, we describe here a novel method for the cultivation of self-regenerating human epithelial cells from small intestinal crypts, which contain both intestinal stem cells and mature villus cells. We present data that suggest these cells support Cryptosporidium better than existing cell lines. PECs should provide an improved tool for studying host-parasite interactions involving Cryptosporidium and other intestinal pathogens. PMID:23509153

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

    Science.gov (United States)

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

    2017-02-01

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

  4. Infectivity and pathogenicity of Cryptosporidium andersoni to a novel host, southern multimammate mouse (Mastomys coucha)

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Ondráčková, Z.; Květoňová, Dana; Sak, Bohumil; Vítovec, J.

    2007-01-01

    Roč. 143, 3/4 (2007), s. 229-233 ISSN 0304-4017 R&D Projects: GA ČR GA524/05/0992 Institutional research plan: CEZ:AV0Z60220518 Keywords : Cryptosporidium andersoni * Mastomys coucha * infectivity * pathogenicity * 18S rRNA gene Subject RIV: EG - Zoology Impact factor: 2.016, year: 2007

  5. COMPARISONS OF ELISA AND WESTERN BLOT ASSAYS FOR DETECTION OF CRYPTOSPORIDIUM ANTIBODY

    Science.gov (United States)

    A seroprevalence survey was conducted using ELISA and Western blot (WB) assays for antibody to three Cryptosporidium antigens on 380 blood donors in Jackson County, Oregon. The purpose was to determine if either assay could detect serological evidence of an outbreak which occurre...

  6. First description of Cryptosporidium ubiquitum XIIa subtype family in farmed fur animals

    Czech Academy of Sciences Publication Activity Database

    Kellnerová, K.; Holubová, Nikola; Jandova, Anna; Vejcik, A.; McEvoy, J.; Sak, Bohumil; Kváč, Martin

    2017-01-01

    Roč. 59, JUN (2017), s. 108-113 ISSN 0932-4739 R&D Projects: GA ČR GA15-01090S Institutional support: RVO:60077344 Keywords : Apicomplexa * Chinchillas * Cryptosporidium * gp60 * Foxes * Mink * Nutrias Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine OBOR OECD: Veterinary science Impact factor: 2.581, year: 2016

  7. Inhibition of Retinoblastoma Protein Inactivation

    Science.gov (United States)

    2017-11-01

    CONTRACT NUMBER Inhibition of Retinoblastoma Protein Inactivation 5b. GRANT NUMBER W81XWH-14-1-0329 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S) Seth M...confirmed 108 compounds as giving a dose-response curve with at least 30% inhibition at 10 µM. The flowchart of hit progression is shown on the...Cancer Research Program under Award No. W81XWH-14-1-0329 to S.M.R. Opinions, interpretations, conclusions, and recommendations are those of the author

  8. A comparative study of the disinfection efficacy of H2O2/ferrate and UV/H2O2/ferrate processes on inactivation of Bacillus subtilis spores by response surface methodology for modeling and optimization.

    Science.gov (United States)

    Matin, Atiyeh Rajabi; Yousefzadeh, Samira; Ahmadi, Ehsan; Mahvi, Amirhossein; Alimohammadi, Mahmood; Aslani, Hassan; Nabizadeh, Ramin

    2018-04-03

    Although chlorination can inactivate most of the microorganisms in water but protozoan parasites like C. parvum oocysts and Giardia cysts can resist against it. Therefore, many researches have been conducted to find a novel method for water disinfection. Present study evaluated the synergistic effect of H2O2 and ferrate followed by UV radiation to inactivate Bacillus subtilis spores as surrogate microorganisms. Response surface methodology(RSM) was employed for the optimization for UV/H2O2/ferrate and H2O2/ferrate processes. By using central composite design(CCD), the effect of three main parameters including time, hydrogen peroxide, and ferrate concentrations was examined on process performance. The results showed that the combination of UV, H2O2 and ferrate was the most effective disinfection process in compare with when H2O2 and ferrate were used. This study indicated that by UV/H2O2/ferrate, about 5.2 log reductions of B. subtilis spores was inactivated at 9299 mg/l of H2O2 and 0.4 mg/l of ferrate concentrations after 57 min of contact time which was the optimum condition, but H2O2/ferrate can inactivate B. subtilis spores about 4.7 logs compare to the other process. Therefore, the results of this research demonstrated that UV/H2O2 /ferrate process is a promising process for spore inactivation and water disinfection. Copyright © 2018 Elsevier Ltd. All rights reserved.

  9. β-1,3-Glucan, Which Can Be Targeted by Drugs, Forms a Trabecular Scaffold in the Oocyst Walls of Toxoplasma and Eimeria

    Science.gov (United States)

    Bushkin, G. Guy; Motari, Edwin; Magnelli, Paula; Gubbels, Marc-Jan; Dubey, Jitender P.; Miska, Katarzyna B.; Bullitt, Esther; Costello, Catherine E.; Robbins, Phillips W.; Samuelson, John

    2012-01-01

    ABSTRACT The walls of infectious pathogens, which are essential for transmission, pathogenesis, and diagnosis, contain sugar polymers that are defining structural features, e.g., β-1,3-glucan and chitin in fungi, chitin in Entamoeba cysts, β-1,3-GalNAc in Giardia cysts, and peptidoglycans in bacteria. The goal here was to determine in which of three walled forms of Toxoplasma gondii (oocyst, sporocyst, or tissue cyst) is β-1,3-glucan, the product of glucan synthases and glucan hydrolases predicted by whole-genome sequences of the parasite. The three most important discoveries were as follows. (i) β-1,3-glucan is present in oocyst walls of Toxoplasma and Eimeria (a chicken parasite that is a model for intestinal stages of Toxoplasma) but is absent from sporocyst and tissue cyst walls. (ii) Fibrils of β-1,3-glucan are part of a trabecular scaffold in the inner layer of the oocyst wall, which also includes a glucan hydrolase that has a novel glucan-binding domain. (iii) Echinocandins, which target the glucan synthase and kill fungi, arrest development of the Eimeria oocyst wall and prevent release of the parasites into the intestinal lumen. In summary, β-1,3-glucan, which can be targeted by drugs, is an important component of oocyst walls of Toxoplasma but is not a component of sporocyst and tissue cyst walls. PMID:23015739

  10. Current status and future trends in Cryptosporidium and Giardia epidemiology in Malaysia.

    Science.gov (United States)

    Lim, Y A L; Ahmad, R A; Smith, H V

    2008-06-01

    Cryptosporidium and Giardia are major causes of diarrhoeal diseases of humans worldwide, and are included in the World Health Organisation's 'Neglected Diseases Initiative'. Cryptosporidium and Giardia occur commonly in Malaysian human and non-human populations, but their impact on disease, morbidity and cost of illness is not known. The commonness of contributions from human (STW effluents, indiscriminate defaecation) and non-human (calving, lambing, muck spreading, slurry spraying, pasturing/grazing of domestic animals, infected wild animals) hosts indicate that many Malaysian environments, particularly water and soil, are sufficiently contaminated to act as potential vehicles for the transmission of disease. To gain insight into the morbidity and mortality caused by human cryptosporidiosis and giardiasis, they should be included into differential diagnoses, and routine laboratory testing should be performed and (as for many infectious diseases) reported to a centralised public health agency. To understand transmission routes and the significance of environmental contamination better will require further multidisciplinary approaches and shared resources, including raising national perceptions of the parasitological quality of drinking water. Here, the detection of Cryptosporidium and Giardia should be an integral part of the water quality requirement. A multidisciplinary approach among public health professionals in the water industry and other relevant health- and environment-associated agencies is also required in order to determine the significance of Cryptosporidium and Giardia contamination of Malaysian drinking water. Lastly, adoption of validated methods to determine the species, genotype and subgenotype of Cryptosporidium and Giardia present in Malaysia will assist in developing effective risk assessment, management and communication models.

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

    Directory of Open Access Journals (Sweden)

    Joyce Siwila

    2012-02-01

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

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

    Science.gov (United States)

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

    2015-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Gabriela Certad

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

  14. Molecular characterization of Cryptosporidium and Giardia from the Tasmanian devil (Sarcophilus harrisii).

    Science.gov (United States)

    Wait, Liana F; Fox, Samantha; Peck, Sarah; Power, Michelle L

    2017-01-01

    The Tasmanian devil (Sarcophilus harrisii) is a carnivorous marsupial found only in the wild in Tasmania, Australia. Tasmanian devils are classified as endangered and are currently threatened by devil facial tumour disease, a lethal transmissible cancer that has decimated the wild population in Tasmania. To prevent extinction of Tasmanian devils, conservation management was implemented in 2003 under the Save the Tasmanian Devil Program. This study aimed to assess if conservation management was altering the interactions between Tasmanian devils and their parasites. Molecular tools were used to investigate the prevalence and diversity of two protozoan parasites, Cryptosporidium and Giardia, in Tasmanian devils. A comparison of parasite prevalence between wild and captive Tasmanian devils showed that both Cryptosporidium and Giardia were significantly more prevalent in wild devils (p Giardia was identified in 24.1% of wild devils but only 0.82% of captive devils. Molecular analysis identified the presence of novel genotypes of both Cryptosporidium and Giardia. The novel Cryptosporidium genotype was 98.1% similar at the 18S rDNA to Cryptosporidium varanii (syn. C. saurophilum) with additional samples identified as C. fayeri, C. muris, and C. galli. Two novel Giardia genotypes, TD genotype 1 and TD genotype 2, were similar to G. duodenalis from dogs (94.4%) and a Giardia assemblage A isolate from humans (86.9%). Giardia duodenalis BIV, a zoonotic genotype of Giardia, was also identified in a single captive Tasmanian devil. These findings suggest that conservation management may be altering host-parasite interactions in the Tasmanian devil, and the presence of G. duodenalis BIV in a captive devil points to possible human-devil parasite transmission.

  15. Re-description of a genetically typed, single oocyst line of the turkey coccidium, Eimeria dispersa Tyzzer, 1929.

    Science.gov (United States)

    El-Sherry, S; Ogedengbe, M E; Hafeez, M A; Sayf-Al-Din, M; Gad, N; Barta, J R

    2017-10-01

    The Briston strain of Eimeria dispersa Tyzzer, 1929 was isolated originally from a commercial turkey flock from Briston, Norfolk, UK. A single oocyst-derived line of E. dispersa was propagated and used to re-describe biological and morphological features of E. dispersa in the turkey. Oocysts of the Briston strain measured 26 ± 1.1 μm (24-28) by 21 ± 1 μm (19-23); these were larger than oocysts described originally by Tyzzer in 1929 (22.75 by 18.84 μm) but within dimensions (26.07 by 21.04 μm) reported by Hawkins (1952) in his description of E. dispersa isolated from turkeys. In the present study, endogenous development started mainly in duodenum and upper jejunum and then spread down toward the lower jejunum. A few parasites were detected in the ileum beginning 96 h post-infection; only few gamonts were observed in the cecal neck area at 120 h, and no parasites were detected in cecal pouches or rectum. Four asexual generations were observed before the start of gametogony, and only one large type of first generation meront was detected in duodenum and upper jejunum at 32 h. This strain has a prepatent period of 120 h. The Briston strain of E. dispersa is a mildly pathogenic coccidium. Duodenum and jejunum of infected birds were slightly dilated and paler in color than of uninfected controls. There was whitish green mucoid material in the lumen of the duodenum and jejunum. The mucosa looked slightly congested and edematous with a few scattered petechial hemorrhages.

  16. Oocyst-Derived Extract of Toxoplasma Gondii Serves as Potent Immunomodulator in a Mouse Model of Birch Pollen Allergy.

    Directory of Open Access Journals (Sweden)

    Angelika Wagner

    Full Text Available Previously, we have shown that oral infection with Toxoplasma gondii oocysts prevented type I allergy in mice. Here we investigated whether the application of a T. gondii oocyst lysate antigen (OLA could also reduce allergy development. BALB/c mice were immunised twice with OLA followed by sensitisation with the major birch pollen (BP allergen Bet v 1 and an aerosol challenge with BP extract.First, we tested OLA in vitro. Stimulation of splenocytes and bone marrow-derived dendritic cells (BMDC with OLA led to the production of pro-inflammatory and regulatory cytokines such as IL-6, IFN-γ and IL-10. Moreover, BMDC exposed to OLA upregulated the maturation markers CD40, CD80, CD86, and MHCII. Furthermore, OLA was recognised by TLR2-transfected human embryonic kidney cells.Immunisation of mice with OLA induced high levels of Toxoplasma-specific IgG antibodies in sera along with increased production of IFN-γ and IL-10 in Toxoplasma-antigen restimulated splenocytes. OLA reduced allergic airway inflammation as manifested by significant reduction of eosinophils in bronchoalveolar fluids, decreased cellular infiltrates and mucus production in the lungs. Accordingly, Bet v 1-specific IgE was decreased in OLA-pretreated mice. The reduced allergic immune responses were accompanied by increased numbers of CD4+CD25highFoxp3+ regulatory T cells in spleens as well as by increased numbers of granulocytic myeloid-derived suppressor cells in lungs when compared to sensitised controls suggesting that these two cell populations might be involved in the suppression of the allergic immune responses.Our data demonstrate that pretreatment with the oocyst extract can exert anti-allergic effects comparable to T. gondii infection. Thus, the immunomodulatory properties of the parasite extract indicate that this extract and in the future defined molecules thereof might serve as immunomodulatory adjuvants in allergy treatment and prophylaxis.

  17. Cryptosporidium tyzzeri and Cryptosporidium muris originated from wild West-European house mice (Mus musculus domesticus) and East-European house mice (Mus musculus musculus) are non-infectious for pigs

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Kestřánová, M.; Květoňová, Dana; Kotková, M.; Ortega, Y.; McEvoy, J.; Sak, Bohumil

    2012-01-01

    Roč. 131, č. 1 (2012), s. 107-110 ISSN 0014-4894 R&D Projects: GA MŠk(CZ) LH11061 Grant - others:GA ČR(CZ) GA206/08/0640 Program:GA Institutional support: RVO:60077344 Keywords : Pigs * Cryptosporidium tyzzeri * Cryptosporidium muris * Experimental infection * PCR * Histology Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.154, year: 2012 http://www.sciencedirect.com/science/article/pii/S0014489412001105

  18. Cryptosporidium meleagridis infection: the first report in Poland of its occurrence in an HIV-positive woman

    Czech Academy of Sciences Publication Activity Database

    Wesołowska, M.; Szostakowska, B.; Kicia, M.; Sak, Bohumil; Kváč, Martin; Knysz, B.

    2016-01-01

    Roč. 62, č. 3 (2016), s. 239-241 ISSN 2299-0631 Institutional support: RVO:60077344 Keywords : Cryptosporidium meleagridis * HIV * AIDS * opportunistic parasites * immunocompromised patients Subject RIV: FN - Epidemiology, Contagious Diseases ; Clinical Immunology

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

    Science.gov (United States)

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

    2017-01-17

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

  20. Epidemiology and molecular characterization of Cryptosporidium spp. in humans, wild primates, and domesticated animals in the Greater Gombe Ecosystem, Tanzania.

    Science.gov (United States)

    Parsons, Michele B; Travis, Dominic; Lonsdorf, Elizabeth V; Lipende, Iddi; Roellig, Dawn M; Roellig, Dawn M Anthony; Collins, Anthony; Kamenya, Shadrack; Zhang, Hongwei; Xiao, Lihua; Gillespie, Thomas R

    2015-02-01

    Cryptosporidium is an important zoonotic parasite globally. Few studies have examined the ecology and epidemiology of this pathogen in rural tropical systems characterized by high rates of overlap among humans, domesticated animals, and wildlife. We investigated risk factors for Cryptosporidium infection and assessed cross-species transmission potential among people, non-human primates, and domestic animals in the Gombe Ecosystem, Kigoma District, Tanzania. A cross-sectional survey was designed to determine the occurrence and risk factors for Cryptosporidium infection in humans, domestic animals and wildlife living in and around Gombe National Park. Diagnostic PCR revealed Cryptosporidium infection rates of 4.3% in humans, 16.0% in non-human primates, and 9.6% in livestock. Local streams sampled were negative. DNA sequencing uncovered a complex epidemiology for Cryptosporidium in this system, with humans, baboons and a subset of chimpanzees infected with C. hominis subtype IfA12G2; another subset of chimpanzees infected with C. suis; and all positive goats and sheep infected with C. xiaoi. For humans, residence location was associated with increased risk of infection in Mwamgongo village compared to one camp (Kasekela), and there was an increased odds for infection when living in a household with another positive person. Fecal consistency and other gastrointestinal signs did not predict Cryptosporidium infection. Despite a high degree of habitat overlap between village people and livestock, our results suggest that there are distinct Cryptosporidium transmission dynamics for humans and livestock in this system. The dominance of C. hominis subtype IfA12G2 among humans and non-human primates suggest cross-species transmission. Interestingly, a subset of chimpanzees was infected with C. suis. We hypothesize that there is cross-species transmission from bush pigs (Potaochoerus larvatus) to chimpanzees in Gombe forest, since domesticated pigs are regionally absent. Our

  1. Molecular characterization of Cryptosporidium and Giardia in farmers and their ruminant livestock from the Coastal Savannah zone of Ghana.

    Science.gov (United States)

    Squire, Sylvia Afriyie; Yang, Rongchang; Robertson, Ian; Ayi, Irene; Ryan, Una

    2017-11-01

    Cryptosporidium and Giardia are major causes of diarrhoea in developing countries including Ghana, however, nothing is known about the species and subtypes of Cryptosporidium and Giardia in farmers and their ruminant livestock in this country. A total of 925 faecal samples from humans (n=95), cattle (n=328), sheep (n=217) and goats (n=285), were screened for Cryptosporidium and Giardia by quantitative PCR (qPCR) at the 18S rRNA and glutamate dehydrogenase (gdh) loci respectively. Cryptosporidium positives were typed by sequence analysis of 18S and 60kDa glycoprotein (gp60) loci amplicons. Giardia positives were typed at the triose phosphate isomerase (tpi), beta-giardin (bg) and gdh loci. The prevalence of Cryptosporidium and Giardia by qPCR was 8.4% and 10.5% in humans, 26.5% and 8.5% in cattle, 34.1% and 12.9% in sheep, and 33.3% and 12.3% in goat faecal samples, respectively. G. duodenalis assemblages A and B were detected in humans and assemblage E was detected in livestock. Cryptosporidium parvum was the only species identified in humans; C. andersoni, C. bovis, C. ryanae and C. ubiquitum were identified in cattle; C. xiaoi, C. ubiquitum and C. bovis in sheep; and C. xiaoi, C. baileyi and C. parvum in goats. This is the first molecular study of Cryptosporidium and Giardia in livestock in Ghana. The identification of zoonotic species and the identification of C. parvum subtype IIcA5G3q in livestock, which has previously been identified in children in Ghana, suggests potential zoonotic transmission. Further studies on larger numbers of human and animal samples, and on younger livestock are required to better understand the epidemiology and transmission of Cryptosporidium and Giardia in Ghana. Copyright © 2017 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

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

  3. Radiobiological inactivation of Epstein-Barr virus

    International Nuclear Information System (INIS)

    Henderson, E.; Heston, L.; Grogan, E.; Miller, G.

    1978-01-01

    Lymphocyte transforming properties of B95-8 strain Epstein-Barr virus (EBV) are very sensitive to inactivation by either uv or x irradiation. No dose of irradiation increases the transforming capacity of EBV. The x-ray dose needed for inactivation of EBV transformation (dose that results in 37% survival, 60,000 rads) is similar to the dose required for inactivation of plaque formation by herpes simplex virus type 1 (Fischer strain). Although herpes simplex virus is more sensitive than EBV to uv irradiation, this difference is most likely due to differences in the kinetics or mechanisms of repair of uv damage to the two viruses. The results lead to the hypothesis that a large part, or perhaps all, of the EBV genome is in some way needed to initiate transformation. The abilities of EBV to stimulate host cell DNA synthesis, to induce nuclear antigen, and to immortalize are inactivated in parallel. All clones of marmoset cells transformed by irradiated virus produce extracellular transforming virus. These findings suggest that the abilities of the virus to transform and to replicate complete progeny are inactivated together. The amounts of uv and x irradiation that inactivate transformation by B95-8 virus are less than the dose needed to inactivate early antigen induction by the nontransforming P 3 HR-1 strain of EBV. Based on radiobiological inactivation, 10 to 50% of the genome is needed for early antigen induction

  4. Photodynamic Inactivation of Mammalian Viruses and Bacteriophages

    Directory of Open Access Journals (Sweden)

    Liliana Costa

    2012-06-01

    Full Text Available Photodynamic inactivation (PDI has been used to inactivate microorganisms through the use of photosensitizers. The inactivation of mammalian viruses and bacteriophages by photosensitization has been applied with success since the first decades of the last century. Due to the fact that mammalian viruses are known to pose a threat to public health and that bacteriophages are frequently used as models of mammalian viruses, it is important to know and understand the mechanisms and photodynamic procedures involved in their photoinactivation. The aim of this review is to (i summarize the main approaches developed until now for the photodynamic inactivation of bacteriophages and mammalian viruses and, (ii discuss and compare the present state of the art of mammalian viruses PDI with phage photoinactivation, with special focus on the most relevant mechanisms, molecular targets and factors affecting the viral inactivation process.

  5. Multilocus typing of Cryptosporidium spp. and Giardia duodenalis from non-human primates in China.

    Science.gov (United States)

    Karim, Md Robiul; Zhang, Sumei; Jian, Fuchun; Li, Jiacheng; Zhou, Chunxiang; Zhang, Longxian; Sun, Mingfei; Yang, Guangyou; Zou, Fengcai; Dong, Haiju; Li, Jian; Rume, Farzana Islam; Qi, Meng; Wang, Rongjun; Ning, Changshen; Xiao, Lihua

    2014-11-01

    Non-human primates (NHPs) are commonly infected with Cryptosporidium spp. and Giardia duodenalis. However, molecular characterisation of these pathogens from NHPs remains scarce. In this study, 2,660 specimens from 26 NHP species in China were examined and characterised by PCR amplification of 18S rRNA, 70kDa heat shock protein (hsp70) and 60kDa glycoprotein (gp60) gene loci for Cryptosporidium; and 1,386 of the specimens by ssrRNA, triosephosphate isomerase (tpi) and glutamate dehydrogenase (gdh) gene loci for Giardia. Cryptosporidium was detected in 0.7% (19/2660) specimens of four NHP species including rhesus macaques (0.7%), cynomolgus monkeys (1.0%), slow lorises (10.0%) and Francois' leaf monkeys (6.7%), belonging to Cryptosporidium hominis (14/19) and Cryptosporidium muris (5/19). Two C. hominis gp60 subtypes, IbA12G3 and IiA17 were observed. Based on the tpi locus, G. duodenalis was identified in 2.2% (30/1,386) of specimens including 2.1% in rhesus macaques, 33.3% in Japanese macaques, 16.7% in Assam macaques, 0.7% in white-headed langurs, 1.6% in cynomolgus monkeys and 16.7% in olive baboons. Sequence analysis of the three targets indicated that all of the Giardia-positive specimens belonged to the zoonotic assemblage B. Highest sequence polymorphism was observed at the tpi locus, including 11 subtypes: three known and eight new ones. Phylogenetic analysis of the subtypes showed that most of them were close to the so-called subtype BIV. Intragenotypic variations at the gdh locus revealed six types of sequences (three known and three new), all of which belonged to so-called subtype BIV. Three specimens had co-infection with C. hominis (IbA12G3) and G. duodenalis (BIV). The presence of zoonotic genotypes and subtypes of Cryptosporidium spp. and G. duodenalis in NHPs suggests that these animals can potentially contribute to the transmission of human cryptosporidiosis and giardiasis. Copyright © 2014 Australian Society for Parasitology Inc. All rights

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

    Science.gov (United States)

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

    2017-03-01

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

  7. Cryptosporidium diagnosis by qPCR in cats at Rio de Janeiro state, Brazil

    Directory of Open Access Journals (Sweden)

    Lara Patrícia Santos Carrasco

    2016-11-01

    Full Text Available ABSTRACT. Carrasco L.P.S., Oliveira R.L.S., Moreira C.M.R., Santos C.R.G.R., Corgozinho K.B. & Souza H.J.M. [Cryptosporidium diagnosis by qPCR in cats at Rio de Janeiro state, Brazil.] Diagnóstico de Cryptosporidium spp. pela técnica de qPCR em gatos no estado do Rio de Janeiro, Brasil. Revista Brasileira de Medicina Veterinária, 38(Supl.:22-26, 2016. Programa de Pós-Graduação em MedicinaVeteriná- ria, Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, BR-465, Km 7, Seropédica, RJ 23890-000, Brasil. E-mail: carrasco.lara@gmail.com Cryptosporidium spp. is recognized as an important etiologic agent of diarrhea in many countries. The aim of this study was to detect the presence of DNA of the parasite Cryptosporidium spp. in feces of cats with history of chronic diarrhea attended in the Feline Medicine Sector of the Veterinary Hospital of the Federal Rural University of Rio de Janeiro, by the polymerase chain reaction technique in real time (RT-PCR. In this study, 100 animals were admitted, of any breed or sex and from 8 weeks of age. As inclusion criteria, patients had to have diarrhea history for more than three weeks, with little success of clinical response to previously established therapies. From the samples obtained by collecting via washing the animal colon and spontaneous defecation, methods of direct examination of the feces, centrifugal flotation technique and real-time PCR were carried out. Of all cats selected for this study, 10% showed infection by Cryptosporidium spp. Most positive animals were aged over one year (70% and only 30% had up to one year old. Cats were 50% purebred and 50% were domestic short hair cats. The clinical signs presented by these cats at the time of consultation were diarrhea (60% and prolapsed rectum (40%. Four animals had co-infections with other enteropathogens (40%, such as Giardia, Toxocara sp. or Tritrichomonas fetus alone or combined. We concluded that infection by

  8. Inactivation of microorganisms in treated municipal wastewater and biosolids by gamma irradiation

    International Nuclear Information System (INIS)

    2009-01-01

    Increasing growth of the world's population, waste minimization policies and agricultural needs make the recycling of domestic wastewater quite a desirable practice. Factors like environmental and public health risks must be taken into account when considering treated wastewater for field irrigation and biosolids for land application. Pathogens present in wastewater and biosolids may remain active after treatment and there is always a great risk of transmission of infections via consuming crop and vegetables. Therefore it is very important to treat domestic wastewater properly before using it as an irrigation water and as a fertilizer. The work reported herein represents an evaluation of the variations in the population densities of below indicated pathogens monitored during a one year study in Ankara Central Municipal Wastewater Treatment Plant, and the efficiency of gamma irradiation for the inactivation of these important waterborne pathogens. Parasitological investigation Treated wastewater and biosolids - Cryptosporidium sp. - Giardia lamblia - Entamoeba histolytica - Cyclospora cayetanensis - Helminth ova Bacteriological investigation Treated wastewater - Total coliforms - Salmonella sp. - Fecal streptococci - Enterococcus sp. Biosolids - Fecal coliforms - Salmonella sp. (Includes 12 tables, 16 figures)

  9. Skewed X-inactivation in cloned mice

    International Nuclear Information System (INIS)

    Senda, Sho; Wakayama, Teruhiko; Yamazaki, Yukiko; Ohgane, Jun; Hattori, Naka; Tanaka, Satoshi; Yanagimachi, Ryuzo; Shiota, Kunio

    2004-01-01

    In female mammals, dosage compensation for X-linked genes is accomplished by inactivation of one of two X chromosomes. The X-inactivation ratio (a percentage of the cells with inactivated maternal X chromosomes in the whole cells) is skewed as a consequence of various genetic mutations, and has been observed in a number of X-linked disorders. We previously reported that phenotypically normal full-term cloned mouse fetuses had loci with inappropriate DNA methylation. Thus, cloned mice are excellent models to study abnormal epigenetic events in mammalian development. In the present study, we analyzed X-inactivation ratios in adult female cloned mice (B6C3F1). Kidneys of eight naturally produced controls and 11 cloned mice were analyzed. Although variations in X-inactivation ratio among the mice were observed in both groups, the distributions were significantly different (Ansary-Bradley test, P < 0.01). In particular, 2 of 11 cloned mice showed skewed X-inactivation ratios (19.2% and 86.8%). Similarly, in intestine, 1 of 10 cloned mice had a skewed ratio (75.7%). Skewed X-inactivation was observed to various degrees in different tissues of different individuals, suggesting that skewed X-inactivation in cloned mice is the result of secondary cell selection in combination with stochastic distortion of primary choice. The present study is the first demonstration that skewed X-inactivation occurs in cloned animals. This finding is important for understanding both nuclear transfer technology and etiology of X-linked disorders

  10. Review of cryptosporidium and giardia in the eastern part of Europe, 2016

    DEFF Research Database (Denmark)

    Plutzer, Judit; Lassen, Brian; Jokelainen, Pikka

    2018-01-01

    Introduction: This paper reviews the current knowledge and understanding of Cryptosporidium spp. an d Giardia spp. in humans, animals and the environment in 10 countries in the eastern part of Europe: Bosnia and Herzegovina, Croatia, Czech Republic, Estonia, Hungary, Latvia, Poland, Romania, Serbia...... and Slovenia. Methods: Published scientific papers and conference proceedings from the international and local literature, official national health service reports, national databases and doctoral theses in local languages were reviewed to provide an extensive overview on the epidemiology, diagnostics...... and research on these pathogens, as well as analyse knowledge gaps and areas for further research. Results: Cryptosporidium spp. and Giardia spp. were found to be common in eastern Europe, but the results from different countries are difficult to compare because of variations in reporting practices...

  11. Chronic diarrhea caused by Blastocystis hominis and Cryptosporidium sp. in immunocompetent patient-a case report

    Science.gov (United States)

    Andriyani, Y.; Rozi, M. F.; Saragih, R. H.; Darlan, D. M.

    2018-03-01

    Blastocystis hominis and Cryptosporidium sp. are commonly associated with immunocompromised patients. Severe clinical manifestation can be produced by this organism. It varies according to immune status, and subtype of this organism. Unfortunately, we found an immunocompetent patient with chronic diarrhea caused by this organism. A 38- year old male was admitted to Adam Malik General Hospital because of watery diarrhea since four days ago. Administration of fluid replacement was done to this patient, but the frequency of diarrhea did not decrease. Loperamide as anti-spasmodic was also given in each episode of diarrhea. Surprisingly, fecal smear examination revealed that this patient positive for Blastocystis hominis and Cryptosporidium sp. Thus, diarrhea was resolved for four days without giving any anti-parasitic drugs to the patient.

  12. Aural-pharyngeal polyps associated with Cryptosporidium infection in three iguanas (Iguana iguana).

    Science.gov (United States)

    Uhl, E W; Jacobson, E; Bartick, T E; Micinilio, J; Schimdt, R

    2001-03-01

    Cryptosporidium spp. infection was associated with aural-pharyngeal polyps in three iguanas (Iguana iguana). All iguanas were presented for masses protruding from the ear canal, and the disease was characterized by a chronic clinical course. The masses consisted of nests of cystic glands surrounded by abundant fibrous connective tissue and lined by hyperplastic cuboidal to pseudostratified columnar epithelium that was moderately to heavily colonized by cryptosporidial organisms. Electron microscopy revealed that the majority of organisms were trophozoites.

  13. Molecular identification of Cryptosporidium spp. in animal and human hosts from the Czech Republic

    Czech Academy of Sciences Publication Activity Database

    Hajdušek, Ondřej; Ditrich, Oleg; Šlapeta, J.

    2004-01-01

    Roč. 122, č. 3 (2004), s. 183-192 ISSN 0304-4017 R&D Projects: GA AV ČR IBS6022006 Grant - others:GA MŠk1(CZ) 1260/2001 Institutional research plan: CEZ:AV0Z6022909 Keywords : Cryptosporidium * molecular identification * SSU rRNA Subject RIV: EE - Microbiology, Virology Impact factor: 1.445, year: 2004

  14. Infectivity of gastric and intestinal Cryptosporidium species in immunocompetent Mongolian gerbils (Meriones unguiculatus)

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Sak, Bohumil; Květoňová, Dana; Secor, W. E.

    2009-01-01

    Roč. 163, 1/2 (2009), s. 33-38 ISSN 0304-4017 R&D Projects: GA AV ČR KJB500960701; GA ČR GP523/07/P117 Institutional research plan: CEZ:AV0Z60220518 Keywords : Cryptosporidium spp. * Meriones unguiculatus * infectivity Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.278, year: 2009

  15. An audit of Cryptosporidium and Giardia detection in Scottish National Health Service Diagnostic Microbiology Laboratories.

    Science.gov (United States)

    Alexander, C L; Currie, S; Pollock, K; Smith-Palmer, A; Jones, B L

    2017-06-01

    Giardia duodenalis and Cryptosporidium species are protozoan parasites capable of causing gastrointestinal disease in humans and animals through the ingestion of infective faeces. Whereas Cryptosporidium species can be acquired locally or through foreign travel, there is the mis-conception that giardiasis is considered to be largely travel-associated, which results in differences in laboratory testing algorithms. In order to determine the level of variation in testing criteria and detection methods between diagnostic laboratories for both pathogens across Scotland, an audit was performed. Twenty Scottish diagnostic microbiology laboratories were invited to participate with questions on sample acceptance criteria, testing methods, testing rates and future plans for pathogen detection. Reponses were received from 19 of the 20 laboratories representing each of the 14 territorial Health Boards. Detection methods varied between laboratories with the majority performing microscopy, one using a lateral flow immunochromatographic antigen assay, another using a manually washed plate-based enzyme immunoassay (EIA) and one laboratory trialling a plate-based EIA automated with an EIA plate washer. Whereas all laboratories except one screened every stool for Cryptosporidium species, an important finding was that significant variation in the testing algorithm for detecting Giardia was noted with only four laboratories testing all diagnostic stools. The most common criteria were 'travel history' (11 laboratories) and/or 'when requested' (14 laboratories). Despite only a small proportion of stools being examined in 15 laboratories for Giardia (2%-18% of the total number of stools submitted), of interest is the finding that a higher positivity rate was observed for Giardia than Cryptosporidium in 10 of these 15 laboratories. These findings highlight that the underreporting of Giardia in Scotland is likely based on current selection and testing algorithms.

  16. New species of Cryptosporidium Tyzzer, 1907 (Apicomplexa) from amphibian host: morphology, biology and phylogeny

    Czech Academy of Sciences Publication Activity Database

    Jirků, Miloslav; Valigurová, A.; Koudela, Břetislav; Křížek, Jaroslav; Modrý, David; Šlapeta, J.

    2008-01-01

    Roč. 55, č. 2 (2008), s. 81-94 ISSN 0015-5683 R&D Projects: GA ČR GD524/03/H133; GA ČR GA524/05/0992; GA MŠk LC522 Institutional research plan: CEZ:AV0Z60220518 Keywords : Cryptosporidium fragile * new species * Duttaphrynus melanostictus * Host specificity * ultrastructure * global amphibian decline * hylogeny * quarantine Subject RIV: EG - Zoology Impact factor: 1.307, year: 2008

  17. Policy, practice and decision making for zoonotic disease management: water and Cryptosporidium.

    Science.gov (United States)

    Austin, Zoë; Alcock, Ruth E; Christley, Robert M; Haygarth, Philip M; Heathwaite, A Louise; Latham, Sophia M; Mort, Maggie; Oliver, David M; Pickup, Roger; Wastling, Jonathan M; Wynne, Brian

    2012-04-01

    Decision making for zoonotic disease management should be based on many forms of appropriate data and sources of evidence. However, the criteria and timing for policy response and the resulting management decisions are often altered when a disease outbreak occurs and captures full media attention. In the case of waterborne disease, such as the robust protozoa, Cryptosporidium spp, exposure can cause significant human health risks and preventing exposure by maintaining high standards of biological and chemical water quality remains a priority for water companies in the UK. Little has been documented on how knowledge and information is translated between the many stakeholders involved in the management of Cryptosporidium, which is surprising given the different drivers that have shaped management decisions. Such information, coupled with the uncertainties that surround these data is essential for improving future management strategies that minimise disease outbreaks. Here, we examine the interplay between scientific information, the media, and emergent government and company policies to examine these issues using qualitative and quantitative data relating to Cryptosporidium management decisions by a water company in the North West of England. Our results show that political and media influences are powerful drivers of management decisions if fuelled by high profile outbreaks. Furthermore, the strength of the scientific evidence is often constrained by uncertainties in the data, and in the way knowledge is translated between policy levels during established risk management procedures. In particular, under or over-estimating risk during risk assessment procedures together with uncertainty regarding risk factors within the wider environment, was found to restrict the knowledge-base for decision-making in Cryptosporidium management. Our findings highlight some key current and future challenges facing the management of such diseases that are widely applicable to other

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

    Science.gov (United States)

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

    2018-04-12

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

  19. The effects of combining Artemisia annua and Curcuma longa ethanolic extracts in broilers challenged with infective oocysts of Eimeria acervulina and E. maxima

    DEFF Research Database (Denmark)

    Almeida, Gustavo Fonseca; Thamsborg, Stig Milan; Madeira, Alda M.B.N

    2014-01-01

    and oocyst excretion were investigated. Broilers given chemical coccidiostats performed better than all other groups. Broilers given the two highest dosages of the herbal mixture had intermediate lesion scores caused by Eimeria acervulina, which was higher than in broilers given coccidiostats, but less than...

  20. Long-term humoral antibody responses by various serologic tests in pigs orally inoculated with oocysts of four strains of Toxoplasma gondii

    DEFF Research Database (Denmark)

    Dubey, J.P.; Andrews, C.D.; Thulliez, P.

    1997-01-01

    Antibody titers to Toxoplasma gondii were determined in 16 pigs orally inoculated with 1000 or 10000 oocysts of one of the four strains (GT-1, ME-49, TS-2, TC-2) of T. gondii. Pigs were euthanized on postinoculation days 103-875 and their tissues were bioassayed for T. gondii. Antibody titers wer...

  1. Detection and genotyping of Entamoeba histolytica, Entamoeba dispar, Giardia lamblia, and Cryptosporidium parvum by oligonucleotide microarray.

    Science.gov (United States)

    Wang, Zheng; Vora, Gary J; Stenger, David A

    2004-07-01

    Entamoeba histolytica, Giardia lamblia, and Cryptosporidium parvum are the most frequently identified protozoan parasites causing waterborne disease outbreaks. The morbidity and mortality associated with these intestinal parasitic infections warrant the development of rapid and accurate detection and genotyping methods to aid public health efforts aimed at preventing and controlling outbreaks. In this study, we describe the development of an oligonucleotide microarray capable of detecting and discriminating between E. histolytica, Entamoeba dispar, G. lamblia assemblages A and B, and C. parvum types 1 and 2 in a single assay. Unique hybridization patterns for each selected protozoan were generated by amplifying six to eight diagnostic sequences/organism by multiplex PCR; fluorescent labeling of the amplicons via primer extension; and subsequent hybridization to a set of genus-, species-, and subtype-specific covalently immobilized oligonucleotide probes. The profile-based specificity of this methodology not only permitted for the unequivocal identification of the six targeted species and subtypes, but also demonstrated its potential in identifying related species such as Cryptosporidium meleagridis and Cryptosporidium muris. In addition, sensitivity assays demonstrated lower detection limits of five trophozoites of G. lamblia. Taken together, the specificity and sensitivity of the microarray-based approach suggest that this methodology may provide a promising tool to detect and genotype protozoa from clinical and environmental samples.

  2. Physical inactivation and stabilization of sludges

    International Nuclear Information System (INIS)

    Alexandre, D.

    1979-07-01

    High temperature conditioning of sludge is a stabilization process that insures sterilization. Both thermal pasteurization and irradiation are inactivation processes. Viruses and parasites are inactivated at 70-80 0 C. Total bacterial destruction requires higher temperatures and/or detention time. Radio sensitivity of pathogens and pertinent treatment parameters are examined. If sludge is to be land disposed, disinfection requires irradiation doses ranging 500 Krad; if cattle feeding is considered, the required dose is 1 Mrad

  3. Microbial Inactivation by Ultrasound Assisted Supercritical Fluids

    Science.gov (United States)

    Benedito, Jose; Ortuño, Carmen; Castillo-Zamudio, Rosa Isela; Mulet, Antonio

    A method combining supercritical carbon dioxide (SC-CO2) and high power ultrasound (HPU) has been developed and tested for microbial/enzyme inactivation purposes, at different process conditions for both liquid and solid matrices. In culture media, using only SC-CO2, the inactivation rate of E. coli and S. cerevisiae increased with pressure and temperature; and the total inactivation (7-8 log-cycles) was attained after 25 and 140 min of SC-CO2 (350 bar, 36 °C) treatment, respectively. Using SC-CO2+HPU, the time for the total inactivation of both microorganisms was reduced to only 1-2 min, at any condition selected. The SC-CO2+HPU inactivation of both microorganisms was slower in juices (avg. 4.9 min) than in culture media (avg. 1.5 min). In solid samples (chicken, turkey ham and dry-cured pork cured ham) treated with SC-CO2 and SC-CO2+HPU, the inactivation rate of E. coli increased with temperature. The application of HPU to the SC-CO2 treatments accelerated the inactivation rate of E. coli and that effect was more pronounced in treatments with isotonic solution surrounding the solid food samples. The application of HPU enhanced the SC-CO2 inactivation mechanisms of microorganisms, generating a vigorous agitation that facilitated the CO2 solubilization and the mass transfer process. The cavitation generated by HPU could damage the cell walls accelerating the extraction of vital constituents and the microbial death. Thus, using the combined technique, reasonable industrial processing times and mild process conditions could be used which could result into a cost reduction and lead to the minimization in the food nutritional and organoleptic changes.

  4. Mycobacteria inactivation using Engineered Water Nanostructures (EWNS).

    Science.gov (United States)

    Pyrgiotakis, Georgios; McDevitt, James; Gao, Ya; Branco, Alan; Eleftheriadou, Mary; Lemos, Bernardo; Nardell, Edward; Demokritou, Philip

    2014-08-01

    Airborne transmitted pathogens such as Mycobacterium tuberculosis (Mtb) cause serious, often fatal infectious disease with enormous global health implications. Due to their unique cell wall and slow growth, mycobacteria are among the most resilient microbial forms. Herein we evaluate the ability of an emerging, chemical-free, nanotechnology-based method to inactivate M. parafortuitum (Mtb surrogate). This method is based on the transformation of atmospheric water vapor into engineered water nano-structures (EWNS) via electrospray. We demonstrate that the EWNS can interact with and inactivate airborne mycobacteria, reducing their concentration levels significantly. Additionally, EWNS can inactivate M. parafortuitum on surfaces eight times faster than the control. The mechanism of mycobacteria inactivation was also investigated in this study. It was demonstrated that the EWNS effectively deliver the reactive oxygen species, encapsulated during the electrospray process, to the bacteria oxidizing their cell membrane resulting into inactivation. Overall, this is a method with the potential to become an effective intervention technology in the battle against airborne infections. This study demonstrates the feasibility of mycobacterium inactivation in airborne form or on contact surfaces using electrospray activated water nano-structures. Given that the method is free of toxic chemicals, this might become an important tool in the prevention of mycobacterial infections, which are notoriously hard to treat. Copyright © 2014 Elsevier Inc. All rights reserved.

  5. Cell inactivation by heavy charged particles

    Energy Technology Data Exchange (ETDEWEB)

    Blakely, E A [Lawrence Berkeley Lab., CA (United States). Cell and Molecular Biology Div.

    1992-06-01

    The inactivation of cells resulting in lethal or aberrant effects by charged particles is of growing interest. Charged particles at extremely high LET are capable of completely eliminating cell-type and cell-line differences in repair capacity. It is still not clear however whether the repair systems are inactivated, or merely that heavy-ion lesions are less repairable. Studies correlating the particle inactivation dose of radioresistant cells with intact DNA analyzed with pulse field gel electrophoresis and other techniques may be useful, but more experiments are also needed to assess the fidelity of repair. For particle irradiations between 40-100 keV/{mu}m there is however evidence for particle-induced activation of specific genes in mammalian cells, and certain repair processes in bacteria. New data are available on the inactivation of developmental processes in several systems including seeds, and cells of the nematode C. elegans. Future experimental and theoretical modeling research emphasis should focus on exploring particle-induced inactivation of endpoints assessing functionality and not just lethality, and on analyzing molecular damage and genetic effects arising in damage but non-inactivated survivors. The discrete nature of selective types of particle damage as a function of radiation quality indicates the value of accelerated ions as probes of normal and aberrant biological processes. Information obtained from molecular analyses of damage and repair must however be integrated into the context of cellular and tissue functions of the organism. (orig.).

  6. CCp5A protein from Toxoplasma gondii as a serological marker of oocyst-driven infections in humans and domestic animals.

    Directory of Open Access Journals (Sweden)

    Silas Silva Santana

    2015-11-01

    Full Text Available Considering that the current immunoassays are not able to distinguish the infective forms that cause Toxoplasma gondii infection, the present study was carried out to evaluate the reactivity of two recombinant proteins (CCp5A and OWP1 from oocyst/sporozoite, in order to differentiate infections occurring by ingestion of oocysts or tissue cysts. The reactivity of the recombinant proteins was assessed against panels of serum samples from animals (chickens, pigs and mice that were naturally or experimentally infected by different infective stages of the parasite. Also, we tested sera from humans who have been infected by oocysts during a well-characterized toxoplasmosis outbreak, as well as sera from pregnant women tested IgM+/IgG+ for T. gondii, which source of infection was unknown. Only the sporozoite-specific CCp5A protein was able to differentiate the parasite stage that infected chickens, pigs and mice, with specific reactivity for oocyst-infected animals. Furthermore, the CCp5A showed preferential reactivity for recent infection by oocyst/sporozoite in pigs and mice. In humans, CCp5A showed higher reactivity with serum samples from the outbreak, compared with serum from pregnant women. Altogether, these findings demonstrate the usefulness of the CCp5A protein as a new tool to identify the parasite state of T. gondii infection, allowing its application for diagnosis and epidemiological investigations in animals and humans. The identification of parasite infective stage can help to design effective strategies to minimize severe complications in immunocompromised people and, particularly, in pregnant women to prevent congenital infection.

  7. Infection rate of Giardia duodenalis, Cryptosporidium spp. and Enterocytozoon bieneusi in cashmere, dairy and meat goats in China.

    Science.gov (United States)

    Peng, Xian-Qi; Tian, Ge-Ru; Ren, Guan-Jing; Yu, Zheng-Qing; Lok, James Barron; Zhang, Long-Xian; Wang, Xue-Ting; Song, Jun-Ke; Zhao, Guang-Hui

    2016-07-01

    Cryptosporidiosis, microsporidiosis, and giardiasis contribute significantly to the high burden of zoonotic diarrhea worldwide. Goats constitute an important species in animal agriculture by providing cashmere wool, meat, and dairy products for human consumption. However, zoonotic pathogens with the potential to cause morbidity and to degrade production have been reported frequently in goats recently. The present study examined 629 fecal specimens from goats, including 315 cashmere goats, 170 dairy goats and 144 meat goats, in multiple cities of Shaanxi and Henan provinces, northwestern and central China, to investigate the infection rate and species/assemblages/genotypes of Giardia duodenalis, Cryptosporidium spp. and Enterocytozoon bieneusi. Of these samples, 274 (43.6%) were positive for three zoonotic pathogens, including 80 (12.7%), 104 (16.5%) and 179 (28.5%) for G. duodenalis, Cryptosporidium spp. and E. bieneusi, respectively. Infections with G. duodenalis, Cryptosporidium spp. and E. bieneusi existed in meat, dairy and cashmere goats, with the highest infection rate of each pathogen being observed in meat goats. DNA sequencing of the SSU rRNA gene from 104 Cryptosporidium-positive specimens revealed existence of Cryptosporidium xiaoi, and the zoonotic parasites Cryptosporidium parvum and Cryptosporidium ubiquitum. Genotyping of G. duodenalis based on the triosephosphate isomerase (TPI) gene identified parasites from zoonotic assemblage A in four cashmere goats and the animal-adapted assemblage E in a group of 76 goats that included cashmere, dairy and meat animals. Polymorphisms in the ribosomal internal transcribed spacer characterized E. bieneusi genotype CHG1 and a novel genotype named as SX1 in both dairy and cashmere goats, genotypes CHS7 and COSI in meat goats, the genotype CHG2 in dairy goats, and the human-pathogenic genotype BEB6 in dairy and meat goats. This is the first detailed study to compare infection rate of the zoonotic protozoan pathogens

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

    Directory of Open Access Journals (Sweden)

    Miles E Daniels

    2018-01-01

    Full Text Available In many low-income settings, despite improvements in sanitation and hygiene, groundwater sources used for drinking may be contaminated with enteric pathogens such as Cryptosporidium and Giardia, which remain important causes of childhood morbidity. In this study, we examined the contribution of diarrhea caused by Cryptosporidium and Giardia found in groundwater sources used for drinking to the total burden of diarrheal disease among children < 5 in rural India.We studied a population of 3,385 children < 5 years of age in 100 communities of Puri District, Odisha, India. We developed a coupled quantitative microbial risk assessment (QMRA and susceptible-infected-recovered (SIR population model based on observed levels of Cryptosporidium and Giardia in improved groundwater sources used for drinking and compared the QMRA-SIR estimates with independently measured all-cause (i.e., all fecal-oral enteric pathogens and exposure pathways child diarrhea prevalence rates observed in the study population during two monsoon seasons (2012 and 2013. We used site specific and regional studies to inform assumptions about the human pathogenicity of the Cryptosporidium and Giardia species present in local groundwater. In all three human pathogenicity scenarios evaluated, the mean daily risk of Cryptosporidium or Giardia infection (0.06-1.53%, far exceeded the tolerable daily risk of infection from drinking water in the US (< 0.0001%. Depending on which protozoa species were present, median estimates of daily child diarrhea prevalence due to either Cryptosporidium or Giardia infection from drinking water was as high as 6.5% or as low as < 1% and accounted for at least 2.9% and as much as 65.8% of the all-cause diarrhea disease burden measured in children < 5 during the study period. Cryptosporidium tended to account for a greater share of estimated waterborne protozoa infections causing diarrhea than did Giardia. Diarrhea prevalence estimates for waterborne

  9. Cryptosporidium sebagai Indikator Biologi dan Indeks Nsf-Wqi untuk Mengevaluasi Kualitas Air (Studi Kasus: Hulu Sungai Citarum, Kabupaten Bandung

    Directory of Open Access Journals (Sweden)

    Tastaptyani Kurnia Nufutomo

    2017-11-01

    Full Text Available Kualitas air yang menurun di Hulu Sungai Citarum dapat disebabkan oleh banyak faktor. Faktor-faktor tersebut dapat diketahui dari parameter fisika, kimia dan biologi. Parameter biologi yang digunakan untuk mengevaluasi kualitas air adalah  mikroorganisme patogen yang menimbulkan penyakit di sistem pencernaan seperti diare akut, yaitu Coliform dan Cryptosporidium. Tujuan penelitian ini adalah untuk mengetahui status kualitas air di Hulu Sungai Citarum dengan indeks kualitas air NSF-WQI, mengetahui hubungan dan pengaruh parameter fisik dan kimia air terhadap parameter biologi, menentukan faktor utama dari parameter air yang paling berpengaruh dan mengetahui hubungan serta pengaruh faktor utama tersebut terhadap Cryptosporidium. Metode yang digunakan adalah  mengambil sampel di tiap stasiun dengan composite, mengidentifikasi dan analisis Coliform dengan MPN dan identifikasi Crytosporidium dengan Ziehl Neelsen staining, kemudian menganalisis parameter kimia dan fisika dengan indeks NSF-WQI, lalu data tersebut diolah menggunakan metode statistik PCA. Hasil pengukuran kualitas air berdasarkan NSF-WQI adalah kualitas air di Hulu Sungai Citarum termasuk kategori buruk dan medium. Keberadaan Cryptosporidium di Hulu Sungai Citarum disebabkan oleh 2 (dua faktor utama, yaitu faktor pertama terdiri dari DO, turbiditas, NO2, NH4 dan total Colifom, sedangkan faktor kedua terdiri dari TSS, COD dan PO4. Kedua faktor tersebut tidak signifikan dengan keberadaan Cryptosporidium di Hulu Sungai Citarum. Kata kunci: Cryptosporidium, Hulu Sungai Citarum, Indeks NSF-WQI, Kualitas Air

  10. Longitudinal prevalence, oocyst shedding and molecular characterisation of Eimeria species in sheep across four states in Australia.

    Science.gov (United States)

    Yang, Rongchang; Jacobson, Caroline; Gardner, Graham; Carmichael, Ian; Campbell, Angus J D; Ryan, Una

    2014-10-01

    The prevalence of Eimeria in sheep in Australia has not been well described, therefore a quantitative PCR (qPCR) was developed, validated and used to study the prevalence and oocyst concentration in lamb faecal samples at three sampling periods (weaning, post-weaning and pre-slaughter) from eight farms across South Australia, New South Wales, Victoria and Western Australia. A total of 3412 faecal samples were collected from approximately 1182 lambs across the 4 states and screened for the presence of Eimeria using this qPCR at the 18S ribosomal RNA (rRNA) locus. A subset of positives was typed by sequence analysis at the 18S locus. The overall prevalence was 18.1% (95% CI 16.8-19.3%) and of the 616 positives, 118 were successfully genotyped. The prevalence of Eimeria was highest in NSW and peaked at 70% during the post-weaning period. The range of oocyst shedding per gram of faeces (g(-1)) at weaning, post-weaning and pre-slaughter overall across all states was 23-2.1×10(7), 23-1.3×10(7) and 23-2.1×10(5), respectively. Median Eimeria shedding g(-1) was higher during post-weaning (1.1×10(3)) and pre-slaughter (1.1×10(3)) than during weaning (206). The following species were identified: Eimeria crandallis, Eimeria ahsata, Eimeria ovinoidalis, Eimeria weybridgensis and Eimeria cylindrica. Of these, E. crandallis and E. ovinoidalis, the most pathogenic species in sheep were responsible for 58.5% of infections typed. This highlights a need for further research to quantify the production impacts of Eimeria in sheep. Copyright © 2014 Elsevier Inc. All rights reserved.

  11. Inactivation of pathogenic bacteria in food matrices: high pressure processing, photodynamic inactivation and pressure-assisted photodynamic inactivation

    Science.gov (United States)

    Cunha, A.; Couceiro, J.; Bonifácio, D.; Martins, C.; Almeida, A.; Neves, M. G. P. M. S.; Faustino, M. A. F.; Saraiva, J. A.

    2017-09-01

    Traditional food processing methods frequently depend on the application of high temperature. However, heat may cause undesirable changes in food properties and often has a negative impact on nutritional value and organoleptic characteristics. Therefore, reducing the microbial load without compromising the desirable properties of food products is still a technological challenge. High-pressure processing (HPP) can be classified as a cold pasteurization technique, since it is a non-thermal food preservation method that uses hydrostatic pressure to inactivate spoilage microorganisms. At the same time, it increases shelf life and retains the original features of food. Photodynamic inactivation (PDI) is also regarded as promising approach for the decontamination of food matrices. In this case, the inactivation of bacterial cells is achieved by the cytotoxic effects of reactive oxygens species (ROS) produced from the combined interaction of a photosensitizer molecule, light and oxygen. This short review examines some recent developments on the application of HPP and PDI with food-grade photosensitizers for the inactivation of listeriae, taken as a food pathogen model. The results of a proof-of-concept trial of the use of high-pressure as a coadjutant to increase the efficiency of photodynamic inactivation of bacterial endospores is also addressed.

  12. Occurrence of Cryptosporidium and Giardia in domestic animals in peri-urban communities of Kafue district, Zambia

    DEFF Research Database (Denmark)

    Siwila, J.; Phiri, I. G. K.; Enemark, Heidi L.

    2013-01-01

    /148), goats (5.9%; 1/17), dogs (25.0%; 5/20) and ducks (6.7%; 2/30). Diarrhoea was not associated with either infection. Age was also not associated with either infection except in dogs where Giardia infection was only detected in animals aged less than six months (p=0.009). It is concluded from this study......Cryptosporidium spp. and Giardia duodenalis are important parasites infecting a wide range of domestic animals worldwide. The aim of the present study was to determine the occurrence of Cryptosporidium spp. and Giardia parasites in different domestic animals living in close contact with humans...... within rural/semiurban communities in Kafue district in Zambia. A single faecal sample per animal was collected from pigs, goats, dogs, ducks, chickens and pigeons and analysed by Merifluor Cryptosporidium/Giardia immunofluorescence antibody assay for the simultaneous detection of these parasites...

  13. Inactivation of enteroviruses in sewage with ozone

    Energy Technology Data Exchange (ETDEWEB)

    Ivanova, O.E.; Bogdanov, M.V.; Kazantseva, V.A.; Gabrilevskaia, L.N.; Kodkind, G.K.H.

    The study of ozone inactivation of enteroviruses in sewage showed the presence in sewage of suspensions of organic origin and bacterial flora to influence the rate of inactivation. The inactivation rate of poliomyelitis virus in sewage free from organic suspension and bacterial flora was significantly higher than that in sewage containing such suspension and bacterial flora. The inactivation rate of enteroviruses was found not to depend upon the protein and salt composition and pH of sewage or strain appurtenance of viruses. The inactivation rate of enteroviruses directly depended upon the dose of ozone and time of contact with it. Differences in the resistance of different types of poliomyelitis virus, ECHO and Coxsackie viruses to the effect of ozone are likely exist. These differences are manifested within the range of relatively small doses of ozone. E. coli is more resistant to ozone than entero-viruses. The results of laboratory studies were used to choose the regimen of sanitation of urban sewage to be used in technological cycles of industrial enterprises.

  14. Inactivation of Mycobacterium avium with free chlorine.

    Science.gov (United States)

    Luh, Jeanne; Mariñas, Benito J

    2007-07-15

    The inactivation kinetics of Mycobacterium avium with free chlorine was characterized by two stages: an initial phase at a relatively fast rate followed by a slower second stage of pseudo first-order kinetics. The inactivation rate of each stage was approximately the same for all experiments performed at a certain condition of pH and temperature; however, variability was observed for the disinfectant exposure at which the transition between the two stages occurred. This variability was not a function of the initial disinfectant concentration, the initial bacterial density, or the bacterial stock. However, the transition to the second stage varied more significantly at high temperatures (30 degrees C), while lower variability was observed at lower temperatures (5 and 20 degrees C). Experiments conducted at pH values in the range of 6-9 revealed that the inactivation of M. avium was primarily due to hypochlorous acid, with little contribution from hypochlorite ion within this pH range. The inactivation kinetics was represented with a two-population model. The activation energies for the resulting pseudo first-order rate constants for the populations with fast and slow kinetics were 100.3 and 96.5 kJ/mol, respectively. The magnitude of these values suggested that for waters of relatively high pH and low temperatures, little inactivation of M. avium would be achieved within treatment plants, providing a seeding source for distribution systems.

  15. Prevalence of Giardia duodenalis and Cryptosporidium species infections among children and cattle in North Shewa Zone, Ethiopia.

    Science.gov (United States)

    Wegayehu, Teklu; Adamu, Haileeyesus; Petros, Beyene

    2013-09-08

    Giardia and Cryptosporidium are the most common causes of protozoan diarrhea that lead to significant morbidity and mortality worldwide. The purpose of this study was to determine the prevalence of Giardia duodenalis and Cryptosporidium species infections among children and cattle, and to assess the potential risk of zoonotic transmission. This cross-sectional study was conducted between January and April 2009 in Girar Jarso and Dera Districts of North Shewa Zone, Oromia Region, Ethiopia. A total of 768 stool specimens were collected and examined for intestinal parasites using direct wet mount with saline and formalin ether concentration methods. The modified Ziehl-Neelsen staining method was used for the detection of Cryptosporidium species. Statistical analysis was performed using SPSS software version 15. Out of 384 children examined, 53 (13.8%) and 28 (7.3%) were positive for Giardia and Cryptosporidium infections, respectively. Similarly, of the total 384 cattle examined, 9 (2.3%) were positive for Giardia duodenalis and 30 (7.8%) were positive for Cryptosporidium infection. The prevalence of giardiasis was significantly higher among children who had close contact with cattle 33 (18.7%) compared to children who had no contact with cattle 20 (9.6%) (P < 0.05). Higher number of Cryptosporidium infection was also recorded in children who had close contact with cattle 15 (8.5%). Difference in prevalence of giardiasis and cryptosporidiosis among children was not statistically significant between males and females. On the other hand, difference in the prevalence of giardiasis among children was statistically significant between age groups. Higher prevalence of Giardia duodenalis infection detected among children was significantly associated with contact with cattle and manure that the children had. Further analysis using molecular techniques is needed to explain the existence of zoonotic transmission in the study area.

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

    Science.gov (United States)

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

    2018-01-01

    In many low-income settings, despite improvements in sanitation and hygiene, groundwater sources used for drinking may be contaminated with enteric pathogens such as Cryptosporidium and Giardia, which remain important causes of childhood morbidity. In this study, we examined the contribution of diarrhea caused by Cryptosporidium and Giardia found in groundwater sources used for drinking to the total burden of diarrheal disease among children cause (i.e., all fecal-oral enteric pathogens and exposure pathways) child diarrhea prevalence rates observed in the study population during two monsoon seasons (2012 and 2013). We used site specific and regional studies to inform assumptions about the human pathogenicity of the Cryptosporidium and Giardia species present in local groundwater. In all three human pathogenicity scenarios evaluated, the mean daily risk of Cryptosporidium or Giardia infection (0.06-1.53%), far exceeded the tolerable daily risk of infection from drinking water in the US (water was as high as 6.5% or as low as cause diarrhea disease burden measured in children causing diarrhea than did Giardia. Diarrhea prevalence estimates for waterborne Cryptosporidium infection appeared to be most sensitive to assumptions about the probability of infection from ingesting a single parasite (i.e. the rate parameter in dose-response model), while Giardia infection was most sensitive to assumptions about the viability of parasites detected in groundwater samples. Protozoa in groundwater drinking sources in rural India, even at low concentrations, especially for Cryptosporidium, may account for a significant portion of child diarrhea morbidity in settings were tubewells are used for drinking water and should be more systematically monitored. Preventing diarrheal disease burdens in Puri District and similar settings will benefit from ensuring water is microbiologically safe for consumption and consistent and effective household water treatment is practiced.

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

    Science.gov (United States)

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

    2010-01-01

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

  18. Deformability Assessment of Waterborne Protozoa Using a Microfluidic-Enabled Force Microscopy Probe

    NARCIS (Netherlands)

    McGrath, John S.; Quist, Jos; Seddon, James Richard Thorley; Lai, Stanley; Lemay, Serge Joseph Guy; Bridle, Helen L.

    2016-01-01

    Many modern filtration technologies are incapable of the complete removal of Cryptosporidium oocysts from drinking-water. Consequently, Cryptosporidium-contaminated drinking-water supplies can severely implicate both water utilities and consumers. Existing methods for the detection of

  19. Genotypes and subtypes of Cryptosporidium spp. in diarrheic lambs and goat kids in northern Greece.

    Science.gov (United States)

    Papanikolopoulou, Vasiliki; Baroudi, Djamel; Guo, Yaqiong; Wang, Yuanfei; Papadopoulos, Elias; Lafi, Shwakat Q; Abd El-Tawab, Mohamed M; Diakou, Anastasia; Giadinis, Nektarios D; Feng, Yaoyu; Xiao, Lihua

    2018-08-01

    Inconsistent data exist on the distribution of zoonotic Cryptosporidium species and subtypes in sheep and goats in European countries, and few such data are available from Greece. In this study, 280 fecal specimens were collected from 132 diarrheic lambs and 148 diarrheic goat kids aged 4 to 15 days on 15 farms in northern Greece, and examined for Cryptosporidium spp. using microscopy of Ziehl-Neelsen-stained fecal smears. Cryptosporidium spp. in 80 microscopy-positive fecal specimens (39 from lambs and 41 from goat kids) were genotyped by PCR-RFLP analysis of the small subunit rRNA gene and subtyped by sequence analysis the 60 kDa glycoprotein gene. Among the 33 specimens successfully genotyped, C. parvum was found in 32 and C. xiaoi in one. Seven subtypes belonging to two subtype families (IIa and IId) were identified among the 29 C. parvum specimens successfully subtyped, including IIaA14G2R1 (1/29), IIaA15G2R1 (6/29), IIaA20G1R1 (7/29), IIdA14G2 (1/29), IIdA15G1 (9/29), IIdA16G1 (3/29), and IIdA23G1 (2/29). Lambs were more commonly infected with C. parvum IIa subtypes, whereas goat kids were more with IId subtypes. The results illustrate that C. parvum is prevalent in diarrheic lambs and goat kids in northern Greece and these animals could potentially play a role in epidemiology of human cryptosporidiosis. Copyright © 2018 Elsevier B.V. All rights reserved.

  20. Community Laboratory Testing for Cryptosporidium: Multicenter Study Retesting Public Health Surveillance Stool Samples Positive for Cryptosporidium by Rapid Cartridge Assay with Direct Fluorescent Antibody Testing.

    Directory of Open Access Journals (Sweden)

    Dawn M Roellig

    Full Text Available Cryptosporidium is a common cause of sporadic diarrheal disease and outbreaks in the United States. Increasingly, immunochromatography-based rapid cartridge assays (RCAs are providing community laboratories with a quick cryptosporidiosis diagnostic method. In the current study, the Centers for Disease Control and Prevention (CDC, the Association of Public Health Laboratories (APHL, and four state health departments evaluated RCA-positive samples obtained during routine Cryptosporidium testing. All samples underwent "head to head" re-testing using both RCA and direct fluorescence assay (DFA. Community level results from three sites indicated that 54.4% (166/305 of Meridian ImmunoCard STAT! positives and 87.0% (67/77 of Remel Xpect positives were confirmed by DFA. When samples were retested by RCA at state laboratories and compared with DFA, 83.3% (155/186 of Meridian ImmunoCard STAT! positives and 95.2% (60/63 of Remel Xpect positives were confirmed. The percentage of confirmed community results varied by site: Minnesota, 39.0%; New York, 63.9%; and Wisconsin, 72.1%. The percentage of confirmed community results decreased with patient age; 12.5% of community positive tests could be confirmed by DFA for patients 60 years of age or older. The percentage of confirmed results did not differ significantly by sex, storage temperature, time between sample collection and testing, or season. Findings from this study demonstrate a lower confirmation rate of community RCA positives when compared to RCA positives identified at state laboratories. Elucidating the causes of decreased test performance in order to improve overall community laboratory performance of these tests is critical for understanding the epidemiology of cryptosporidiosis in the United States (US.

  1. Genotypes of Cryptosporidium spp., Enterocytozoon bieneusi and Giardia duodenalis in dogs and cats in Shanghai, China.

    Science.gov (United States)

    Xu, Hailing; Jin, Yue; Wu, Wenxian; Li, Pei; Wang, Lin; Li, Na; Feng, Yaoyu; Xiao, Lihua

    2016-03-01

    Controversies exist on the potential role of companion animals in the transmission of enteric pathogens in humans. This study was conducted to examine the genotype distribution of Cryptosporidium spp., Enterocytozoon bieneusi, and Giardia duodenalis in companion animals in Shanghai, China, and to assess their zoonotic potential. Fecal specimens from 485 dogs and 160 cats were examined for the occurrence and genotype distribution of the three pathogens by PCR. PCR products were sequenced to determine the species and genotypes. The χ(2) test was used to compare differences in infection rates between living conditions or age groups. Cryptosporidium spp., E. bieneusi and G. duodenalis were found in 39 (8.0 %), 29 (6.0 %) and 127 (26.2 %) of dogs, and 6 (3.8 %), 9 (5.6 %) and 21 (13.1 %) of cats, respectively. Infection rates of the pathogens in dogs from pet shops and a clinic were higher than those in household dogs, and higher in cats from one animal shelter than from pet shops. No significant differences in infection rates were detected among age groups. Cryptosporidium canis and C. felis were the only Cryptosporidium species found in dogs and cats, respectively. Enterocytozoon bieneusi genotype PtEb IX was the dominant genotype in dogs, whereas Type IV and D were the most common ones in cats. Multi-locus sequence typing at the glutamate dehydrogenase, β-giardin, and triosephosphate isomerase loci revealed the presence of G. duodenalis assemblages A (n = 23), B (n = 1), C (n = 26), and D (n = 58) in dogs (only A in household dogs) and assemblages A (n = 2), B (n = 6), C (n = 2), D (n = 1), and F (n = 7) in cats. Co-infection was detected in 24 dogs and 5 cats, especially those living in crowded conditions. Living condition is a major risk factor affecting the occurrence of enteric protists in companion animals in China, and although dogs and cats can be potential sources of human infections, the different distribution of

  2. Loop-Mediated Isothermal Amplification-Lateral-Flow Dipstick (LAMP-LFD) to detect Toxoplasma gondii oocyst in ready-to-eat salad.

    Science.gov (United States)

    Lalle, Marco; Possenti, Alessia; Dubey, Jitender P; Pozio, Edoardo

    2018-04-01

    The apicomplexan parasite Toxoplasma gondii is the causative agent of toxoplasmosis, a foodborne zoonosis with a global distribution and estimated to cause up to 20% of the total foodborne disease burden in Europe. Association between T. gondii infection and the consumption of unwashed raw fruits and vegetables contaminated with oocysts has been reported and the increasing habit to eat pre-washed ready-to-eat salads poses a new potential risk for consumers. It is therefore important to trace the occurrence of potential contamination with this parasite to guarantee the safety of ready-to-eat vegetables. Detection of T. gondii in vegetables by molecular techniques has been achieved but low sensitivity (PCR) or expensive equipments (qPCR) limit routine applicability. Here, we describe the development and validation of a sensitive and robust method relying on a LAMP assay, targeting the 529 bp locus, to detect T. gondii oocysts down to 25 oocysts/50 g in ready-to-eat baby lettuce. The LAMP has been also adapted for a faster visualization of the result by a lateral flow dipstick chromatographic detection method. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Concentration and retention of Toxoplasma gondii oocysts by marine snails demonstrate a novel mechanism for transmission of terrestrial zoonotic pathogens in coastal ecosystems

    Science.gov (United States)

    Krusor, Colin; Smith, Woutrina A.; Tinker, M. Tim; Silver, Mary; Conrad, Patricia A.; Shapiro, Karen

    2015-01-01

    The parasite Toxoplasma gondii is an environmentally persistent pathogen that can cause fatal disease in humans, terrestrial warm-blooded animals and aquatic mammals. Although an association between T. gondii exposure and prey specialization on marine snails was identified in threatened California sea otters, the ability of kelp-dwelling snails to transmit terrestrially derived pathogens has not been previously investigated. The objective of this study was to measure concentration and retention of T. gondii by marine snails in laboratory aquaria, and to test for natural T. gondii contamination in field-collected snails. Following exposure to T. gondii-containing seawater, oocysts were detected by microscopy in snail faeces and tissues for 10 and 3 days respectively. Nested polymerase chain reaction was also applied as a method for confirming putative T. gondii oocysts detected in snail faeces and tissues by microscopy. Toxoplasma gondiiwas not detected in field-collected snails. Results suggest that turban snails are competent transport hosts for T. gondii. By concentrating oocysts in faecal pellets, snails may facilitate entry of T. gondii into the nearshore marine food web. This novel mechanism also represents a general pathway by which marine transmission of terrestrially derived microorganisms can be mediated via pathogen concentration and retention by benthic invertebrates.

  4. High Pressure Inactivation of HAV within Mussels

    Science.gov (United States)

    The potential of hepatitis A virus (HAV) to be inactivated within Mediterranean mussels (Mytilus galloprovincialis) and blue mussels (Mytilus edulis) by high pressure processing was evaluated. HAV was bioaccumulated within mussels to approximately 6-log10 PFU by exposure of mussels to HAV-contamina...

  5. Inactivation of prion infectivity by ionizing rays

    Energy Technology Data Exchange (ETDEWEB)

    Gominet, M. [Ionisos, ZI les Chatinieres, F01120 Dagneux (France); Vadrot, C.; Austruy, G. [Paris V University, Central Pharmacy of Hospitals, 4 avenue de l' Observatoire, F-75006, Paris (France); Darbord, J.C. [Paris V University, Central Pharmacy of Hospitals, 4 avenue de l' Observatoire, F-75006, Paris (France)], E-mail: darbord@pharmacie.univ-paris5.fr

    2007-11-15

    Inactivation of prion deposits on medical devices or prion contamination in pharmaceutical raw materials is considered as impossible by using gamma irradiation. Early, the guideline WHO/CDS/CSR/APH/2000 has described irradiation as an ineffective process. But, in 2003, S. Miekka et al. noted radiation inactivation of prions in a particular application to purify human albumin, shown by the physical denaturation of the infectious protein (PrP). The aim of our study was to determine the inactivation of prions with a scrapie model (strain C506M3) by irradiating standardised preparations. Results: Gamma irradiation was partially effective, showing a 4-5 log reduction on exposure to 50 kGy. A characteristic effect-dose curve was not observed (25, 50 and 100 kGy), only an increase in the incubation period of the murine disease (229 days with 25 kGy to 290 days with 100 kGy) compared with 170 days without irradiation. Since the inactivation was not a total one, the observed effect is significant. It is proposed that further work be undertaken with the model to investigate the application of gamma radiation known levels of prion contamination.

  6. Inactivation of prion infectivity by ionizing rays

    International Nuclear Information System (INIS)

    Gominet, M.; Vadrot, C.; Austruy, G.; Darbord, J.C.

    2007-01-01

    Inactivation of prion deposits on medical devices or prion contamination in pharmaceutical raw materials is considered as impossible by using gamma irradiation. Early, the guideline WHO/CDS/CSR/APH/2000 has described irradiation as an ineffective process. But, in 2003, S. Miekka et al. noted radiation inactivation of prions in a particular application to purify human albumin, shown by the physical denaturation of the infectious protein (PrP). The aim of our study was to determine the inactivation of prions with a scrapie model (strain C506M3) by irradiating standardised preparations. Results: Gamma irradiation was partially effective, showing a 4-5 log reduction on exposure to 50 kGy. A characteristic effect-dose curve was not observed (25, 50 and 100 kGy), only an increase in the incubation period of the murine disease (229 days with 25 kGy to 290 days with 100 kGy) compared with 170 days without irradiation. Since the inactivation was not a total one, the observed effect is significant. It is proposed that further work be undertaken with the model to investigate the application of gamma radiation known levels of prion contamination

  7. Pulsed electric field inactivation in a microreactor

    NARCIS (Netherlands)

    Fox, M.B.

    2006-01-01

    Pulsed electric fields (PEF) is a novel, non-thermal pasteurization method which uses short, high electric field pulses to inactivate microorganisms. The advantage of a pasteurization method like PEF compared to regular heat pasteurization is that the taste, flavour, texture and nutritional value

  8. Epigenetic inactivation of CHFR in human tumors.

    Science.gov (United States)

    Toyota, Minoru; Sasaki, Yasushi; Satoh, Ayumi; Ogi, Kazuhiro; Kikuchi, Takefumi; Suzuki, Hiromu; Mita, Hiroaki; Tanaka, Nobuyuki; Itoh, Fumio; Issa, Jean-Pierre J; Jair, Kam-Wing; Schuebel, Kornel E; Imai, Kohzoh; Tokino, Takashi

    2003-06-24

    Cell-cycle checkpoints controlling the orderly progression through mitosis are frequently disrupted in human cancers. One such checkpoint, entry into metaphase, is regulated by the CHFR gene encoding a protein possessing forkhead-associated and RING finger domains as well as ubiquitin-ligase activity. Although defects in this checkpoint have been described, the molecular basis and prevalence of CHFR inactivation in human tumors are still not fully understood. To address this question, we analyzed the pattern of CHFR expression in a number of human cancer cell lines and primary tumors. We found CpG methylation-dependent silencing of CHFR expression in 45% of cancer cell lines, 40% of primary colorectal cancers, 53% of colorectal adenomas, and 30% of primary head and neck cancers. Expression of CHFR was precisely correlated with both CpG methylation and deacetylation of histones H3 and H4 in the CpG-rich regulatory region. Moreover, CpG methylation and thus silencing of CHFR depended on the activities of two DNA methyltransferases, DNMT1 and DNMT3b, as their genetic inactivation restored CHFR expression. Finally, cells with CHFR methylation had an intrinsically high mitotic index when treated with microtubule inhibitor. This means that cells in which CHFR was epigenetically inactivated constitute loss-of-function alleles for mitotic checkpoint control. Taken together, these findings shed light on a pathway by which mitotic checkpoint is bypassed in cancer cells and suggest that inactivation of checkpoint genes is much more widespread than previously suspected.

  9. The Lesser Egyptian Gerbil (Gerbillus gerbillus) is a suitable host for the long-term propagation of Cryptosporidium andersoni

    Czech Academy of Sciences Publication Activity Database

    Kváč, Martin; Ondráčková, Z.; Květoňová, Dana; McEvoy, J.; Vitovec, J.; Rost, M.; Sak, Bohumil

    2013-01-01

    Roč. 134, č. 4 (2013), s. 438-442 ISSN 0014-4894 R&D Projects: GA MŠk(CZ) LH11061 Grant - others:JČU(CZ) 011/2013/Z Institutional support: RVO:60077344 Keywords : Cryptosporidium andersoni * Infection dynamics * Transmission * Gerbils Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.859, year: 2013

  10. Risk Factors and Spatial Clusters of Cryptosporidium Infection among School-Age Children in a Rural Region of Eastern China.

    Science.gov (United States)

    Zheng, Hao; He, Jianfeng; Wang, Li; Zhang, Rong; Ding, Zhen; Hu, Wenbiao

    2018-05-06

    The epidemiological features of Cryptosporidium infection among school-age children in China still remain unclear. Hereby, a cross-sectional study of 1637 children aged 3⁻9 years was designed to investigate the risk factors and spatial clusters of Cryptosporidium infection in a rural region of Eastern China. Stool specimens collected from participants were examined using the auramine-phenol and modified acid-fast staining. Univariable and multivariable analyses were performed to identify the risk factors of Cryptospordium infection. The spatial clusters were analyzed by a discrete Poisson model using SaTScan software. Our results showed that the overall prevalence of Cryptosporidium infection was 11‰ in the research region. At the age of 3⁻6 years (odds ratios (OR) = 3.072, 95% confidence intervals (CI) : 1.001⁻9.427), not washing hands before eating and after defecation (OR = 3.003, 95% CI: 1.060⁻8.511) were recognized as risk factors. Furthermore, a high-risk spatial cluster (relative risk = 4.220, p = 0.025) was identified. These findings call for effective sustainable interventions including family and school-based hygienic education to reduce the prevalence of Cryptosporidium infection. Therefore, an early warning system based spatiotemporal models with risk factors is required to further improve the effectiveness and efficiency of cryptosporidiosis control in the future.

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

    Directory of Open Access Journals (Sweden)

    Jonatas Campos Almeida

    Full Text Available The purpose of this study was to investigate the occurrence of Cryptosporidium spp. and Giardia spp. in a public water-treatment system. Samples of raw and treated water were collected and concentrated using the membrane filtration technique. Direct Immunofluorescence Test was performed on the samples. DNA extraction using a commercial kit was performed and the DNA extracted was submitted to a nested-PCR reaction (n-PCR and sequencing. In the immunofluorescence, 2/24 (8.33% samples of raw water were positive for Giardia spp.. In n-PCR and sequencing, 2/24 (8.33% samples of raw water were positive for Giardia spp., and 2/24 (8.33% samples were positive for Cryptosporidium spp.. The sequencing showed Cryptosporidium parvum and Giardia duodenalis DNA. In raw water, there was moderate correlation among turbidity, color and Cryptosporidium spp. and between turbidity and Giardia spp.. The presence of these protozoans in the water indicates the need for monitoring for water-treatment companies.

  12. Prevalence of Cryptosporidium Infection in Immunocompromised Pa-tients, In South-West of Iran, 2009-10

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    A Balouty Dehkordy

    2010-12-01

    Full Text Available Background: Cryptosporidium is a protozoan parasite with worldwide distribution. The aim of this study was to estimate the prevalence of Cryptosporidium infection by antigen detection in faeces among immunocompromised patients referred to educational hospitals of Ahvaz City, South-West of Iran, 2009-2010.Methods: Fecal samples from 176 immunocompromised patients were collected and Cryptosporid­ium coproantigen test was performed using ELISA method (DRG kit, Germany. A questionnaire was completed for each case and the results were analyzed using descriptive and Chi-Square tests, by SPSS statistical software (15th version.Results: Our study indicated 5.1% Cryptosporidium infection prevalence in the immunocompro­mised participated population. Furthermore, 4.2 %, 4%, 4.5 % and 9.1% infection rates were identi­fied in children suffered from hematopoietic malignancy, adult cancer patients, renal trans­plant recipients, and HIV+ cases, respectively. There was not significant correlation between the infection and age and gender (P>0.05. Infection was most frequent among HIV+ patients.Conclusion: The present study confirmed the high prevalence of Cryptosporidium antigen in fe­cal samples of immunocompromised patients in the region. As no chemotherapeutic agents have yet proven, especially in immunosuppressed patients, therefore our results highlight the impor­tance of preventive intervention in these groups.

  13. Prevalence of Giardia and Cryptosporidium in beef cows in southern Ontario and in beef calves in southern British Columbia.

    Science.gov (United States)

    McAllister, Tim A; Olson, Merle E; Fletch, Andy; Wetzstein, Merv; Entz, Toby

    2005-01-01

    In 1998 and 1999, fecal samples were collected from 669 beef cows on 39 farms located within 10 counties of Ontario. Overall prevalences of Giardia, Cryptosporidium muris, and Cryptosporidium parvum in cows were 8.7%, 10.6%, and 18.4%, respectively. Of the 39 farms sampled, Giardia was detected on 64%, Cr. muris on 72%, and Cr. parvum on 90%. Cryptosporidium parvum was detected in 28% of the cows in 1998 and in 5.2% in 1999. Differences between the 2 y were attributed to sampling during calving in 1998 and during gestation in 1999. In 1998, Giardia, Cr. muris, and Cr. parvum were detected in herds provided with municipal water. In 1998, 193 calves were sampled from 10 farms, representing 4 watersheds, in British Columbia. Thirty-six percent of the calves exhibited signs of diarrhea. Overall prevalences of Giardia and Cryptosporidium spp. in calves were 36% and 13%, respectively. There was evidence that calves with Giardia were more likely to develop scours. Restricting cattle from surface water during periods of high shedding may reduce watershed contamination.

  14. Highly divergent 18S rRNA gene paralogs in a Cryptosporidium genotype from eastern chipmunks (Tamias striatus)

    Czech Academy of Sciences Publication Activity Database

    Stenger, B.L.S.; Clark, M.E.; Kváč, Martin; Khan, E.; Giddings, C.W.; Dyer, N.W.; Schultz, J.L.; McEvoy, J.M.

    2015-01-01

    Roč. 32, JUN 2015 (2015), s. 113-123 ISSN 1567-1348 R&D Projects: GA MŠk(CZ) LH11061 Institutional support: RVO:60077344 Keywords : Cryptosporidium * Paralogy * 18S rRNA * 18S rDNA Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.591, year: 2015

  15. Effect of Cryptosporidium parvum infection on the absorptive capacity and paracellular permeability of the small intestine in neonatal calves

    Czech Academy of Sciences Publication Activity Database

    Klein, P.; Kleinová, T.; Volek, Z.; Šimůnek, Jiří

    2008-01-01

    Roč. 152, 1-2 (2008), s. 53-59 ISSN 0304-4017 Institutional research plan: CEZ:AV0Z50450515 Keywords : calves * cryptosporidium parvum * intestinal absorption Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.039, year: 2008

  16. Cortical inactivation by cooling in small animals

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    Ben eCoomber

    2011-06-01

    Full Text Available Reversible inactivation of the cortex by surface cooling is a powerful method for studying the function of a particular area. Implanted cooling cryoloops have been used to study the role of individual cortical areas in auditory processing of awake-behaving cats. Cryoloops have also been used in rodents for reversible inactivation of the cortex, but recently there has been a concern that the cryoloop may also cool non-cortical structures either directly or via the perfusion of blood, cooled as it passed close to the cooling loop. In this study we have confirmed that the loop can inactivate most of the auditory cortex without causing a significant reduction in temperature of the auditory thalamus or other sub-cortical structures. We placed a cryoloop on the surface of the guinea pig cortex, cooled it to 2°C and measured thermal gradients across the neocortical surface. We found that the temperature dropped to 20-24°C among cells within a radius of about 2.5mm away from the loop. This temperature drop was sufficient to reduce activity of most cortical cells and led to the inactivation of almost the entire auditory region. When the temperature of thalamus, midbrain, and middle ear were measured directly during cortical cooling, there was a small drop in temperature (about 4°C but this was not sufficient to directly reduce neural activity. In an effort to visualise the extent of neural inactivation we measured the uptake of thallium ions following an intravenous injection. This confirmed that there was a large reduction of activity across much of the ipsilateral cortex and only a small reduction in subcortical structures.

  17. Occurrence of Cryptosporidium spp. in two centers of training horses in Curitiba, Paraná

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    Keila Youko Fujii

    2014-09-01

    Full Text Available O presente trabalho investigou a ocorrência de parasitismo por Cryptosporidium spp. em equinos alojados em dois centros de treinamento de equinos localizados no município de Curitiba, Paraná. Foram examinados 108 cavalos, sendo 48 procedentes do Centro de Treinamento 1 (CT1 e 60 do Centro de Treinamento 2 (CT2. As coletas de amostras de fezes foram realizadas no período de outubro de 2010 a janeiro de 2011. A metodologia utilizada para a confirmação da presença de oocistos de Cryptosporidium spp. foi a técnica de Ziehl-Neelsen modificada. A ocorrência encontrada foi de 18,52% para o total de animais examinados. Houve diferença estatística significativa (p > 0,05 quando comparadas as prevalências encontradas nos dois centros de treinamento, sendo no CT1 de 4,16% e no CT2 de 30%. Não houve associação entre a prevalência e a idade, o sexo e raça (p > 0,05.

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

    Science.gov (United States)

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

    2014-03-01

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

  19. Occurrence of Campylobacter spp. and Cryptosporidium spp. in seagulls (Larus spp.).

    Science.gov (United States)

    Moore, John E; Gilpin, Deidre; Crothers, Elizabeth; Canney, Anne; Kaneko, Aki; Matsuda, Motoo

    2002-01-01

    An investigation was carried out into the prevalence of thermophilic Campylobacter subspecies (spp.) and Cryptosporidium spp. in fresh fecal specimens collected from members of the gull family (Larus spp.) from three coastal locations of Northern Ireland. A total of 205 fresh fecal specimens were collected from gulls, of which 28 of 205 (13.7%) were positive for Campylobacter spp. and none of 205 for Cryptosporidium spp. Of these campylobacters, 21 of 28 (75%) isolates obtained belonged to the urease-positive thermophilic Campylobacter (UPTC) taxon, followed by five of 28 (17.9%) Campylobacter lari and 2/28 (7.1%) Campylobacter jejuni. It is significant that seagulls are the sole warm-blooded animal host of this bacterial taxon in Northern Ireland. It is proposed that physiological adaptation to starvation by gulls may lead to increased concentrations of urea through energy production from protein, yielding increased levels of urea for metabolism by UPTC organisms. In general, the possibility exists that environmental contamination of surface waters with campylobacters might be mediated by wild birds (such as gulls), where such waters are used for recreational purposes or where such waters are consumed untreated, might represent a risk to public health.

  20. Effect of climate change on runoff of Campylobacter and Cryptosporidium from land to surface water.

    Science.gov (United States)

    Sterk, Ankie; Schijven, Jack; de Roda Husman, Ana Maria; de Nijs, Ton

    2016-05-15

    Faeces originating from wildlife, domestic animals or manure-fertilized fields, is considered an important source of zoonotic pathogens to which people may be exposed by, for instance, bathing or drinking-water consumption. An increase in runoff, and associated wash-off of animal faeces from fields, is assumed to contribute to the increase of disease outbreaks during periods of high precipitation. Climate change is expected to increase winter precipitation and extreme precipitation events during summer, but has simultaneously also other effects such as temperature rise and changes in evapotranspiration. The question is to what extent the combination of these effects influence the input of zoonotic pathogens to the surface waters. To quantitatively analyse the impacts of climate change on pathogen runoff, pathogen concentrations reaching surface waters through runoff were calculated by combining an input model for catchment pathogen loads with the Wageningen Lowland Runoff Simulator (WALRUS). Runoff of Cryptosporidium and Campylobacter was evaluated under different climate change scenarios and by applying different scenarios for sources of faecal pollution in the catchments, namely dairy cows and geese and manure fertilization. Model evaluation of these scenarios shows that climate change has little overall impact on runoff of Campylobacter and Cryptosporidium from land to the surface waters. Even though individual processes like runoff fluxes, pathogen release and dilution are affected, either positively or negatively, the net effect on the pathogen concentration in surface waters and consequently also on infection risks through recreation seems limited. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.