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Sample records for oocysts

  1. Plasmodium Oocysts: Overlooked Targets of Mosquito Immunity.

    Science.gov (United States)

    Smith, Ryan C; Barillas-Mury, Carolina

    2016-12-01

    Although the ability of mosquitoes to limit Plasmodium infection is well documented, many questions remain as to how malaria parasites are recognized and killed by the mosquito host. Recent evidence suggests that anti-Plasmodium immunity is multimodal, with different immune mechanisms regulating ookinete and oocyst survival. However, most experiments determine the number of mature oocysts, without considering that different immune mechanisms may target different developmental stages of the parasite. Complement-like proteins have emerged as important determinants of early immunity targeting the ookinete stage, yet the mechanisms by which the mosquito late-phase immune response limits oocyst survival are less understood. Here, we describe the known components of the mosquito immune system that limit oocyst development, and provide insight into their possible mechanisms of action. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. 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...

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

    Directory of Open Access Journals (Sweden)

    Elahe Ebrahimzade

    2014-12-01

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

  4. The Effects of Exogenous Nitric Oxide on Eimeria tenella Oocysts

    Institute of Scientific and Technical Information of China (English)

    LI Jin-gui; ZHU Bei-lei; JIANG Jin-shu

    2003-01-01

    The effects of exogenous NO on Eimeria tenella oocysts were studied. The E. tenella oocystsfreshly isolated from feces of experimental chickens infected with E. tenella and sporulated E. tenella oocystswere treated in vitro by the well-known NO-donors, SNP, SPER/NO and GSNO. The results showed thatamong the 3 NO donors, only GSNO had significant effects on the percentage of sporulated oocysts, the inhibi-ting rate of purified oocysts was from 93 to 97%, and reached 100% on the oocysts in the feces of chickenswith E. tenella infection. GSNO also significantly decreased the virulence of E. tenella oocysts, and the oo-cysts treated by 2 -8 mmol L-1 GSNO almost lost their pathogenicity to chickens.

  5. 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

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

    DEFF Research Database (Denmark)

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

    2012-01-01

    . Among leachate samples containing oocysts, 44/72 samples yielded viable oocysts as determined by a dye permeability assay (DAPI/PI) with the majority (41%) of viable oocysts found in leachate from soil columns with added liquid slurry. The number of viable oocysts was positively correlated (r=0...

  7. 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.

  8. 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.

  9. 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.

  10. A novel procedure for total nucleic acid extraction from small numbers of Eimeria species oocysts.

    Science.gov (United States)

    Kaya, Galip; Dale, Colin; Maudlin, Ian; Morgan, Kenton

    2007-01-01

    A series of experiments were performed in an attempt to extract genomic DNA from a small number of Eimerian oocysts. Sonication, ammonia, ethanol and lysozyme were all found to be unsuitable for the digestion of Eimeria oocysts. The chemicals and enzyme given were not capable of either disruption or digestion of oocysts for nucleic acid extraction. They had the capability of penetrating the oocyst wall but could not break-up the oocyst wall. It is impossible to obtain nucleic acid from Eimeria oocysts if the wall is not broken-up. In this study oocyst disruption was achieved using a simple but highly effective treatment regime involving sodium hypochlorite treatment, osmotic shock and proteinase K digestion. Following the disruption of the oocyst walls, a commercially available nucleic acid purification kit (Wizard DNA Purification Kit, Promega) can be used to prepare high quality nucleic acid.

  11. 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.

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

    OpenAIRE

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

    1990-01-01

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

  13. Methods to produce and safely work with large numbers of Toxoplasma gondii oocysts and bradyzoite cysts

    OpenAIRE

    H. Fritz; Barr, B.; Packham, A.; Melli, A.; Conrad, P A

    2011-01-01

    Two major obstacles to conducting studies with Toxoplasma gondii oocysts are the difficulty in reliably producing large numbers of this life stage and safety concerns because the oocyst is the most environmentally resistant stage of this zoonotic organism. Oocyst production requires oral infection of the definitive feline host with adequate numbers of T. gondii organisms to obtain unsporulated oocysts that are shed in the feces for 3-10 days after infection. Since the most successful and comm...

  14. Determining UV Inactivation of Toxoplasma gondii Oocysts by Using Cell Culture and a Mouse Bioassay

    Science.gov (United States)

    The effect of UV exposure on Toxoplasma gondii oocysts has not been completely defined for use in water disinfection. This study evaluated UV irradiated oocysts by three assays: a SCID mouse bioassay, an in vitro T. gondii oocyst plaque assay (TOP-assay), and a quantitative reve...

  15. Methods to produce and safely work with large numbers of Toxoplasma gondii oocysts and bradyzoite cysts.

    Science.gov (United States)

    Fritz, H; Barr, B; Packham, A; Melli, A; Conrad, P A

    2012-01-01

    Two major obstacles to conducting studies with Toxoplasma gondii oocysts are the difficulty in reliably producing large numbers of this life stage and safety concerns because the oocyst is the most environmentally resistant stage of this zoonotic organism. Oocyst production requires oral infection of the definitive feline host with adequate numbers of T. gondii organisms to obtain unsporulated oocysts that are shed in the feces for 3-10 days after infection. Since the most successful and common mode of experimental infection of kittens with T. gondii is by ingestion of bradyzoite tissue cysts, the first step in successful oocyst production is to ensure a high bradyzoite tissue cyst burden in the brains of mice that can be used for the oral inoculum. We compared two methods for producing bradyzoite brain cysts in mice, by infecting them either orally or subcutaneously with oocysts. In both cases, oocysts derived from a low passage T. gondii Type II strain (M4) were used to infect eight-ten week-old Swiss Webster mice. First the number of bradyzoite cysts that were purified from infected mouse brains was compared. Then to evaluate the effect of the route of oocyst inoculation on tissue cyst distribution in mice, a second group of mice was infected with oocysts by one of each route and tissues were examined by histology. In separate experiments, brains from infected mice were used to infect kittens for oocyst production. Greater than 1.3 billion oocysts were isolated from the feces of two infected kittens in the first production and greater than 1.8 billion oocysts from three kittens in the second production. Our results demonstrate that oral delivery of oocysts to mice results in both higher cyst loads in the brain and greater cyst burdens in other tissues examined as compared to those of mice that received the same number of oocysts subcutaneously. The ultimate goal in producing large numbers of oocysts in kittens is to generate adequate amounts of starting material

  16. 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

    2016-01-01

    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. PMID:27641141

  17. Immunization with Eimeria ninakohlyakimovae-live attenuated oocysts protect goat kids from clinical coccidiosis.

    Science.gov (United States)

    Ruiz, Antonio; Muñoz, María Carmen; Molina, José Manuel; Hermosilla, Carlos; Andrada, Marisa; Lara, Pedro; Bordón, Elisa; Pérez, Davinia; López, Adassa María; Matos, Lorena; Guedes, Aránzazu Carmen; Falcón, Soraya; Falcón, Yaiza; Martín, Sergio; Taubert, Anja

    2014-01-17

    Caprine coccidiosis, affecting mainly young goat kids around the weaning period, is worldwide the most important disease in the goat industry. Control of caprine coccidiosis is increasingly hampered by resistances developed against coccidiostatic drugs leading to an enhanced need for anticoccidial vaccines. In the current study we conducted an oral immunization trial with live attenuated sporulated Eimeria ninakohlyakimovae oocysts. Sporulated E. ninakohlyakimovae oocysts were attenuated by X-irradiation technique. The experimental design included a total of 18 goat kids divided into the following groups: (i) animals immunized with attenuated E. ninakohlyakimovae oocysts at 5 weeks of age and challenged 3 weeks later with non-irradiated homologous oocysts (group 1); (ii) animals infected with non-attenuated E. ninakohlyakimovae oocysts at 5 weeks of age and challenged 3 weeks later with non-attenuated homologous oocysts (group 2); (iii) animals primary-infected with untreated E. ninakohlyakimovae oocysts at 8 weeks of age (control of the challenge infection, group 3); (iv) non-infected control animals (group 4). Goat kids immunized with live attenuated E. ninakohlyakimovae oocysts (group 1) excreted significantly less oocysts in the faeces (95.3% reduction) than kids infected with non-attenuated ones (group 2). Furthermore, immunization with live but attenuated oocysts resulted in ameliorated clinical coccidiosis compared to goat kids infected with untreated oocysts (group 2) and resulted in equally reduced signs of coccidiosis after challenge infection compared to acquired immunity driven by non-attenuated oocysts. Overall, the present study demonstrates for the first time that live attenuated E. ninakohlyakimovae oocysts orally administered showed almost no pathogenicity but enough immunogenicity in terms of immunoprotection. Importantly, vaccinated animals still shed low amounts of oocysts, guaranteeing environmental contamination and consecutive booster

  18. Schizogony and gametogony of oocyst-deficient T-263 strain of Toxoplasma gondii.

    Science.gov (United States)

    Dubey, J P

    2017-05-27

    Oocysts are important stage for the spread of Toxoplasma gondii because they are environmentally resistant. Among all hosts of T. gondii, only felids can excrete oocysts. Cats that have excreted T. gondii oocysts after primary infection develop immunity to re-excretion of oocysts, and this immunity appears to be long-lasting. It would be desirable to have a non-infectious vaccine for the prevention of T. gondii infection in cats and to understand mechanism of immunity to excretion of oocysts. An initial step will be to indentify stage/stages of the parasite for induction of immunity. A chemically-induced mutant of T. gondii, T-263, is immunogenic but lacks the capacity to form oocysts in cats. Cats fed live bradyzoites of T-263 do not excrete oocysts after challenge with oocyst producing strains. However, it is not known at what stage of the parasite development the oocyst formation is halted. Here, four cats were fed live tissue cysts of the T-263 strain and examined for enteroepithelial stages and oocyst production. Two cats were administered methyl prednisolone aceatate (20mg/kg) once intramuscularly and these cats were euthanized 5 and 7days post inoculation. No oocysts but immature and mature schizonts (types D and E), male, and female gamonts were detected in two cats euthanized. The remaining two cats did not excrete oocysts examined 3-14days post inoculation, but both seroconverted and developed antibody titers of 1:400 tested by the modification agglutination test, indicating exposure to the inocula. The results demonstrate that the T-263 strain is defective in oocyst formation and the observations should help future studies in identification of genes/factors responsible for oocyst formation in the intestine of cats. Published by Elsevier B.V.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1990-05-01

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

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

    Science.gov (United States)

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

    1990-01-01

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

  1. Toxoplasma gondii: uptake and survival of oocysts in free-living amoebae.

    Science.gov (United States)

    Winiecka-Krusnell, Jadwiga; Dellacasa-Lindberg, Isabel; Dubey, J P; Barragan, Antonio

    2009-02-01

    Waterborne transmission of the oocyst stage of Toxoplasma gondii can cause outbreaks of clinical toxoplasmosis in humans and infection of marine mammals. In water-related environments and soil, free-living amoebae are considered potential carriers of various pathogens, but knowledge on interactions with parasitic protozoa remains elusive. In the present study, we assessed whether the free-living Acanthamoeba castellanii, due to its phagocytic activity, can interact with T. gondii oocysts. We report that amoebae can internalize T. gondii oocysts by active uptake. Intracellular oocysts in amoebae rarely underwent phagocytic lysis, retained viability and established infection in mice. Interaction of T. gondii with amoebae did not reduce the infectivity and pathogenicity of oocysts even after prolonged co-cultivation. Our results show that uptake of oocysts by A. castellanii does not restrain the transmission of T. gondii in a murine infection model.

  2. Exit of Plasmodium sporozoites from oocysts is an active process that involves the circumsporozoite protein.

    Directory of Open Access Journals (Sweden)

    2005-09-01

    Full Text Available Plasmodium sporozoites develop within oocysts residing in the mosquito midgut. Mature sporozoites exit the oocysts, enter the hemolymph, and invade the salivary glands. The circumsporozoite (CS protein is the major surface protein of salivary gland and oocyst sporozoites. It is also found on the oocyst plasma membrane and on the inner surface of the oocyst capsule. CS protein contains a conserved motif of positively charged amino acids: region II-plus, which has been implicated in the initial stages of sporozoite invasion of hepatocytes. We investigated the function of region II-plus by generating mutant parasites in which the region had been substituted with alanines. Mutant parasites produced normal numbers of sporozoites in the oocysts, but the sporozoites were unable to exit the oocysts. In in vitro as well, there was a profound delay, upon trypsin treatment, in the release of mutant sporozoites from oocysts. We conclude that the exit of sporozoites from oocysts is an active process that involves the region II-plus of CS protein. In addition, the mutant sporozoites were not infective to young rats. These findings provide a new target for developing reagents that interfere with the transmission of malaria.

  3. Development of a Cryptosporidium oocyst assay using an automated fiber optic-based biosensor

    Directory of Open Access Journals (Sweden)

    Kramer Marianne F

    2007-10-01

    Full Text Available Abstract An intestinal protozoan parasite, Cryptosporidium parvum, is a major cause of waterborne gastrointestinal disease worldwide. Detection of Cryptosporidium oocysts in potable water is a high priority for the water treatment industry to reduce potential outbreaks among the consumer populace. Anti-Cryptosporidium oocyst polyclonal and monoclonal antibodies were tested as capture and detection reagents for use in a fiber optic biosensor assay for the detection of Cryptosporidium oocysts. Antibodies were validated using enzyme-linked immunosorbent assays, flow cytometry, Western blotting and fluorescent microscopy. Oocysts could be detected at a concentration of 105 oocysts/ml when the polyclonal antibodies were used as the capture and detection reagents. When oocysts were boiled prior to detection, a ten-fold increase in sensitivity was achieved using the polyclonal antibody. Western blotting and immunofluorescence revealed that both the monoclonal and polyclonal antibodies recognize a large (>300 kDa molecular weight mucin-like antigen present on the surface of the oocyst wall. The polyclonal antibody also reacted with a small (105 kDa molecular weight antigen that was present in boiled samples of oocysts. Preliminary steps to design an in-line biosensor assay system have shown that oocysts would have to be concentrated from water samples and heat treated to allow detection by a biosensor assay.

  4. Experimental Toxoplasma gondii oocyst infections in turkeys (Meleagris gallopavo).

    Science.gov (United States)

    Bangoura, B; Zöller, B; Koethe, M; Ludewig, M; Pott, S; Fehlhaber, K; Straubinger, R K; Daugschies, A

    2013-09-23

    Toxoplasma (T.) gondii is a protozoan parasite with a broad range of intermediate hosts. Humans are often infected by ingestion of tissue cysts in raw or undercooked meat or meat products. Turkeys as food-producing animals can also serve as intermediate hosts. The aim of the present study was to investigate occurrence and predilection sites of T. gondii infection in turkeys after oral infection with oocysts. Experimental infections with different doses of T. gondii oocysts were performed in 36 turkeys to mimic natural infection. Systemic distribution of parasitic stages was investigated by screening 14 different tissues including the edible tissues heart, liver, thigh, breast and drumstick muscle. Parasite detection was based on a conventional nested polymerase chain reaction (PCR). Animals were sacrificed 6-12 weeks after infection. Results demonstrated parasite spreading over the whole organism after oral infection by oocysts. Most frequently affected tissues were brain (47.2% of all brains were positive for T. gondii) and thigh muscle (25.0% positive samples). Other muscles were regularly T. gondii-positive, all other sampled tissues were positive at least once. Thus, edible tissues are one of the predilection sites of T. gondii in turkeys which renders raw or undercooked turkey meat a potential risk for parasite transmission to humans. Data were compared to results from previous parenteral turkey infections with tachyzoites. With the exception of brain, liver and breast muscle affection, no significant differences were observed between both infection routes. Both infection models could be used for research purposes with certain advantages and disadvantages.

  5. Evaluation of an experimental irradiated oocyst vaccine to protect broiler chicks against avian coccidiosis.

    Science.gov (United States)

    Fetterer, Raymond H; Jenkins, Mark C; Miska, Katarzyna B; Barfield, Ruth C

    2014-09-01

    The current study investigates the use of irradiated oocysts to protect broiler chicks, raised on litter, from infection with multiple species of Eimeria. In order to determine the optimum radiation dose for each Eimeria species, 1-day-old chicks were immunized with oocysts of Eimeria maxima, Eimeria acervulina, or Eimeria tenella exposed to gamma radiation ranging from 0-500 Gy. The litter oocyst counts at 7 days postimmunization, and the effect on weight gain following a challenge infection, decreased with an optimum dose between 150-200 Gy. Based on this finding, broiler chicks were immunized with a mixture of E. maxima, E. acervulina, and E tenella that had been exposed to 150 or 200 Gy. This resulted in more than a 100-fold reduction in litter oocyst counts and significant protection from a challenge infection, as measured by improved weight gain and feed conversion ratio (FCR). Immunization of birds with oocysts receiving 200 Gy was less effective in providing protection from a challenge infection. An additional formulation of vaccines containing two different oocyst doses of the three species that had been irradiated with 150 Gy were evaluated in their ability to attenuate oocyst output and convey protection to challenge. Results were similar with both high and low numbers of irradiated oocysts. Immunized chicks shed less oocysts at 7 days postimmunization and were protected from negative effects of challenge infection as measured by FCR, changes in weight gain, lesion scores, and measurement of body composition. However, the level of protection was somewhat less than that achieved by immunization with nonirradiated oocysts. The overall conclusion is that an irradiated oocyst vaccine developed in this study can effectively protect chicks that are raised on litter from challenge infection with multiple species of Eimeria, comparable to vaccines with virulent or precocious strains.

  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. Viability staining of Cryptosporidium oocysts and Giardia cysts combined with flow cytometry

    NARCIS (Netherlands)

    Schets FM; Medema GJ; MGB

    1998-01-01

    The incorporation of flow cytometry as an additional purification step has improved the detection method for Cryptosporidium oocysts and Giardia cysts in water. Flow cytometry allows separation of (oo)cysts from interfering debris particles present in water concentrates and thus enables the applicat

  8. Increased efficacy of Eimeria oocysts delivery by gel beads or spray vaccination

    Science.gov (United States)

    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 E. acervulina, E. maxima, and E. tenella oocysts as a vaccine to protect ...

  9. Effects of ozone and UV radiation treatments on the infectivity of Toxoplasma gondii oocysts

    Science.gov (United States)

    Clinical toxoplasmosis in humans has been epidemiologically-linked to the consumption of drinking water contaminated by Toxoplasma gondii oocysts. We evaluated killing of T. gondii oocysts after ultraviolet (UV) or ozone treatments by bioassay in mice and/or cell culture. A 4-log inactivation of the...

  10. 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...

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

    Science.gov (United States)

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

    2014-05-20

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

  12. Effects of ozone and ultraviolet radiation treatments on the infectivity of Toxoplasma gondii oocysts.

    Science.gov (United States)

    Dumètre, Aurélien; Le Bras, Caroline; Baffet, Maxime; Meneceur, Pascale; Dubey, J P; Derouin, Francis; Duguet, Jean-Pierre; Joyeux, Michel; Moulin, Laurent

    2008-05-31

    Clinical toxoplasmosis in humans has been epidemiologically linked to the consumption of drinking water contaminated by Toxoplasma gondii oocysts. We evaluated killing of T. gondii oocysts after ultraviolet (UV) or ozone treatments by bioassay in mice and/or cell culture. A 4-log inactivation of the oocyst/sporozoite infectivity was obtained for UV fluences >20 mJ cm(-2). In contrast, oocysts were not inactivated by ozone with an exposure (Ct) up to 9.4 mg min l (-1) in water at 20 degrees C. In conclusion, UV treatment can be an effective disinfection method to inactivate T. gondii oocysts in drinking water, but ozone did not show promise in this research.

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

    Science.gov (United States)

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

    1997-01-01

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

  14. Gel-Bead Delivery of Eimeria oocysts protects chickens against coccidiosis.

    Science.gov (United States)

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

    2012-06-01

    Vaccines composed of either virulent or attenuated Eimeria spp. oocysts have been developed as an alternative to medication of feed with ionophore drugs or synthetic chemicals. The purpose of this study was to evaluate the use of gel-beads containing a mixture of Eimeria acervulina, Eimeria maxima, and Eimeria tenella oocysts as a vaccine against coccidiosis. Newly hatched chicks (Gallus gallus domesticus) were either sprayed with an aqueous suspension of Eimeria oocysts or were allowed to ingest feed containing Eimeria oocysts-incorporated gel-beads. Control day-old chicks were given an equivalent number of Eimeria oocysts (10(4) total) by oral gavage. After 3 days, chicks were randomly assigned to individual cages, and feces were collected between days 5 and 8 postinfection. All samples were processed for total Eimeria oocysts. 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. Oocyst excretion by chicks fed gel-beads or inoculated by oral gavage was 10- to 100-fold greater than that of chicks spray-vaccinated with the Eimeria oocysts mixture (log 6.3-6.6 vs. log 4.8). Subsequent protection against challenge as measured by weight gain and feed conversion efficiency was significantly greater (P 0.05) in weight gain and feed conversion efficiency compared with nonchallenged controls. These findings indicate that incorporation of Eimeria spp. oocysts in gel-beads may represent an effective way to deliver live oocyst vaccines to day-old chicks for preventing subsequent outbreaks of coccidiosis in the field.

  15. Experimental infection of dogs (Canis familiaris) with sporulated oocysts of Neospora caninum.

    Science.gov (United States)

    Bandini, Luciana A; Neto, Aldo F A; Pena, Hilda F J; Cavalcante, Guacyara T; Schares, Gereon; Nishi, Sandra M; Gennari, Solange M

    2011-03-10

    Neospora caninum is widely distributed in the world and this parasite is one of the major causes of abortion in cattle. Dogs and coyotes are definitive hosts of N. caninum and several species of domestic and wild animals are intermediate hosts. Dogs can become infected by the ingestion of tissues containing cysts and then excrete oocysts. It is not yet known whether sporulated oocysts are able to induce a patent infection in dogs, i.e. a shedding of N. caninum oocysts in feces. The objective of this study was to experimentally examine the infection of dogs by sporulated oocysts. The oocysts used in the experiment were obtained by feeding dogs with brain of buffaloes (Bubalus bubalis) positive for anti-N. caninum antibodies by indirect fluorescent antibody test (IFAT ≥200). Oocysts shed by these dogs were confirmed to be N. caninum by molecular methods and by bioassay in gerbils, and sporulated N. caninum oocysts were used for the oral infection of four dogs. The dogs were 8 weeks old and negative for antibodies to N. caninum and Toxoplasma gondii. Dogs 1 and 4 received an inoculum of 10,000 sporulated oocysts each; dog 2 an inoculum of 5000 sporulated oocysts and dog 3 received 1000 sporulated oocysts of N. caninum. The total feces excreted by these dogs were collected and examined daily for a period of 30 days. No oocysts were found in their feces. The dogs were monitored monthly for a 6-month period to observe a possible seroconversion and when this occurred the animals were eliminated from the experiment. Dogs 1 and 4 seroconverted 1 month after the infection with titer, in the IFAT, of 1600 and 800, respectively; the other two dogs presented no seroconvertion during the 6-month period. Dogs 1 and 2 were euthanized 180 days after infection and were examined for the detection of N. caninum in tissues (brain, muscle, lymph node, liver, lung, heart and bone marrow) by immunohistochemistry and PCR with negative results in both techniques. Bioassay in gerbils with

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

    Science.gov (United States)

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

    2001-04-01

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

  17. 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

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

    OpenAIRE

    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; Meier, Paul

    2011-01-01

    Undetected contamination of food and water by oocysts causes human infections in North America. Risks often are unrecognized. Education alone cannot prevent suffering and economic consequences associated with congenital toxoplasmosis. Prenatal screening can facilitate prevention and treatment of congenital toxoplasmosis.

  19. EVALUATION OF CRYPTOSPORIDIUM OOCYSTS AND GIARDIA CYSTS IN A WATERSHED RESERVOIR

    Science.gov (United States)

    This investigation evaluated the occurrence of Cryptosporidium oocysts and Giardia cysts at 17 sampling locations in Lake Texoma reservoir using method 1623 with standard Envirocheck™ capsule filters. The watershed serves rural agricultural communities active in cattle ranching, ...

  20. Occurrence of Cryptosporidiumspp. oocysts in low quality water and on vegetables in Kumasi, Ghana

    DEFF Research Database (Denmark)

    Petersen, Tobias; Petersen, Heidi Huus; 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....... contamination level of river water, irrigation water and lettuce, 10L of water and 16 lettuce samples were collected four times in the period of, September – October 2013, with weekly intervals from six sample sites in and around Kumasi, Ghana. Oocysts were purified from water by sedimentation for 2 x 48 hours...... to characterize Cryptosporidium spp. positive samples were done by PCR amplification and sequencing of the SSU rRNA, the HSP70 and the GP60 genes after. Cryptosporidium oocysts were found in 75% of the water samples and on 43% of the lettuce with concentrations of 53 – 3268 per 10 L water and 11 – 118 oocyst per...

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

    Science.gov (United States)

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

    2001-01-01

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

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

    Science.gov (United States)

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

    2001-01-01

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

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

    Science.gov (United States)

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

    2001-07-01

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

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

    Science.gov (United States)

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

    2016-11-01

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

  5. 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.

  6. Land use and soil contamination with Toxoplasma gondii oocysts in urban areas.

    Science.gov (United States)

    Gao, Xiang; Wang, Hongbin; Wang, Huan; Qin, Hongyu; Xiao, Jianhua

    2016-10-15

    Because soil contaminated with Toxoplasma gondii oocysts is increasingly recognized as a major source of infection for humans, in this study, we investigated the spatial pattern of soil contamination with T. gondii oocysts in urban area of northeastern Mainland China. From April 2014 to May 2015, more than 9000 soil samples were collected. Detection of T. gondii oocysts was performed applying real-time quantitative PCR. Sensitivity was improved by analyzing four replicates for each sampling point. T. gondii was detected in 30.3% of all samples. Subsequently, a maximum entropy model was used to evaluate the effect of land use and intrinsic soil properties on the risk of contamination with oocysts. Jackknife analysis revealed that the likelihood for positive results is significantly enhanced in soil originating from foci of human habitation, wood land and grass land. Furthermore, soil temperature and humidity significantly influence the probability of contamination with T. gondii oocysts. Our findings indicate that land use may affect distribution of T. gondii oocysts in urban areas.

  7. Transport of Cryptosporidium oocysts in porous media: Role of straining and physicochemical filtration

    Science.gov (United States)

    Tufenkji, N.; Miller, G.F.; Ryan, J.N.; Harvey, R.W.; Elimelech, M.

    2004-01-01

    The transport and filtration behavior of Cryptosporidium parvum oocysts in columns packed with quartz sand was systematically examined under repulsive electrostatic conditions. An increase in solution ionic strength resulted in greater oocyst deposition rates despite theoretical predictions of a significant electrostatic energy barrier to deposition. Relatively high deposition rates obtained with both oocysts and polystyrene latex particles of comparable size at low ionic strength (1 mM) suggest that a physical mechanism may play a key role in oocyst removal. Supporting experiments conducted with latex particles of varying sizes, under very low ionic strength conditions where physicochemical filtration is negligible, clearly indicated that physical straining is an important capture mechanism. The results of this study indicate that irregularity of sand grain shape (verified by SEM imaging) contributes considerably to the straining potential of the porous medium. Hence, both straining and physicochemical filtration are expected to control the removal of C. parvum oocysts in settings typical of riverbank filtration, soil infiltration, and slow sand filtration. Because classic colloid filtration theory does not account for removal by straining, these observations have important implications with respect to predictions of oocyst transport.

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

    Science.gov (United States)

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

    2013-01-01

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

  9. 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

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

    Science.gov (United States)

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

    2010-01-01

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

  11. Oocyst output and transmission rates during successive infections with Eimeria acervulina in experimental broiler flocks.

    Science.gov (United States)

    Velkers, Francisca C; Bouma, Annemarie; Stegeman, J Arjan; de Jong, Mart C M

    2012-06-08

    The infection dynamics of Eimeria species determine the clinical manifestation of the disease coccidiosis in poultry flocks, and a better understanding of the dynamics may contribute to improvement of control measures. Our aim was to study the course of infection and the transmission of Eimeria acervulina in groups of broilers by quantifying the transmission rate parameter and oocyst output. Three transmission experiments were carried out with groups of 20 male SPF broilers. At 2 days of age, one bird in each trial was orally inoculated with five sporulated E. acervulina oocysts (D0 post-inoculation, pi). One day after inoculation (D1 pi), the inoculated bird was housed with 19 non-inoculated contact birds. Individual faecal droppings were examined daily from D3-D32 pi to quantify the number of oocysts per gram faeces. The inoculated bird started shedding oocysts at D5 pi and contact birds between D10 and D17 pi. Contact birds that became infected due to oocyst excretion by the inoculated bird were characterized as first generation contact birds (C1). Contact birds excreting from D15 pi onwards (C2) became infected after the first C1 birds had started shedding and were considered to belong to a successive generation of the flock infection. Oocyst output was significantly lower for C1 compared to C2 birds, but the transmission rate parameter remained constant for both infection generations. These results suggest that although oocyst load increases, the transmission rate of E. acervulina remains constant between successive generations of infection in a flock.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1977-04-01

    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.

  13. Mini-FLOTAC for counting Toxoplasma gondii oocysts from cat feces--comparison with cell counting plates.

    Science.gov (United States)

    Djokic, Vitomir; Blaga, Radu; Rinaldi, Laura; Le Roux, Delphine; Ducry, Tamara; Maurelli, Maria Paola; Perret, Catherine; Djurkovic Djakovic, Olgica; Cringoli, Giuseppe; Boireau, Pascal

    2014-12-01

    Oocysts of Toxoplasma gondii represent one of the most common environmental contaminants causing the zoonotic infection toxoplasmosis. The aim of the present study was to compare the Mini-FLOTAC device with traditional cell counting plates (Kova Slide) for the detection of T. gondii oocysts from feline feces. Two types of experiments were performed: (i) purified oocysts were counted in different dilutions and (ii) specific pathogen free T. gondii-negative cat feces was inoculated with numbers of purified oocysts and counting was performed directly from feces. Our analysis showed a thousand times higher sensitivity of Mini-FLOTAC (5 × 10(2) oocysts) compared to Kova Slide (5 × 10(5) oocysts). Also, when compared by McNemar's test, counting of the purified oocysts showed a higher sensitivity of Mini-FLOTAC compared to Kova Slide, for a dilution of 10(3) oocysts/ml (chi(2) = 6.1; P microscopes in any laboratory or field conditions. We therefore recommend its use for regular screening. Further studies are needed to validate Mini-FLOTAC for the detection of oocysts in soil and water samples in field conditions.

  14. Potential cryptosporidium surrogates and evaluation of compressible oocysts

    Energy Technology Data Exchange (ETDEWEB)

    Li, S.Y.; Goodrich, J.A.; Owens, J.H. [Environmental Protection Agency, Cincinnati, OH (United States)] [and others

    1995-10-01

    Cryptosporidium has been recognized as an important waterborne agent of gastroenteritis and a biological contaminant in drinking water. The widespread presence of Cryptosporidium in surface source water and either untreated or insufficiently treated drinking water has led to Cryptosporidium outbreaks in the United States and worldwide. Among the conventional control practices, filtration and high temperature distillation appear to be the potentially viable technologies for protection against Cryptosporidium in drinking water. As employed in many water plants, filtration is likely to be the most practical treatment technology utilized for Cryptosporidium removal in the near future. Consequently, accurate and reliable methods for evaluation of Cryptosporidium removal rates for filtration-based systems are necessary to assist States in determining drinking water quality and complying with the up-coming national standard for Cryptosporidium in drinking water. Furthermore, searching for reliable and non-hazardous surrogates for evaluation of treatment plant efficiency has been intensified because of the potential health risk associated with Cryptosporidium. Additionally, during the filtration procedure Cryptosporidium may squeeze and fold through pores size of the filtration systems that are smaller than the diameter of the organism; a fraction of these Cryptosporidium oocysts may still remain a certain degree of viability. These uncertainties are critical for the evaluation and optimization of filtration-based physical treatment systems. The in-house research studies described below consist of two parts. One is a potential surrogate study using bag filtration systems at the US EPA Test & Evaluation Facility in Cincinnati, Ohio. The second is Cryptosporidium compressibility and viability investigation.

  15. Occurrence of Cryptosporidium oocysts in fecal samples submitted for routine microbiological examination.

    Science.gov (United States)

    Ratnam, S; Paddock, J; McDonald, E; Whitty, D; Jong, M; Cooper, R

    1985-01-01

    During a 7-month period, 2,252 fecal samples submitted for routine microbiological examination from 1,621 patients were screened for Cryptosporidium oocysts by the auramine staining method with Kinyoun acid-fast stain as the confirmatory stain. Cryptosporidium oocysts were detected in fecal samples from 19 (1.2%) patients, 18 of whom had gastroenteritis. Diarrheic stools from 14 of these 18 patients were negative for the usual enteropathogens but contained the oocysts in moderate to large numbers. Although Cryptosporidium oocysts were found in patients of all ages, they occurred slightly more frequently in infants and children than in the rest. Cryptosporidium species was one of the common enteropathogens identified in fecal samples submitted for routine parasitological examination during the period of the survey and was second only to Giardia species in terms of frequency. Considering cryptosporidiosis in the differential diagnosis of gastroenteritis in immunocompetent persons and including a search for Cryptosporidium oocysts in routine parasitological examinations of fecal samples appear warranted. PMID:4044798

  16. Electrical cream separator coupled with vacuum filtration for the purification of eimerian oocysts and trichostronglyid eggs

    Science.gov (United States)

    El-Ashram, Saeed; Suo, Xun

    2017-02-01

    Several methods have been proposed for separation of eimerian oocysts and trichostronglyid eggs from extraneous debris; however, these methods have been considered to be still inconvenient in terms of time and wide-ranging applications. We describe herein an alternative way using the combination of electrical cream separator and vacuum filtration for harvesting and purifying eimerian oocysts and haemonchine eggs on large-scale applications with approximately 81% and 92% recovery rates for oocysts and nematode eggs obtained from avian and ovine faeces, correspondingly. The sporulation percentages as a measure of viability in the harvested oocysts and eggs from dry faecal materials are nearly 68% and 74%, respectively, and 12 liters of faecal suspension can be processed in approximately 7.5 min. The mode of separation in terms of costs (i.e. simple laboratory equipments and comparably cheap reagents) and benefits renders the reported procedure an appropriate pursuit to harvest and purify parasite oocysts and eggs on a large scale in the shortest duration from diverse volumes of environmental samples compared to the modified traditional sucrose gradient, which can be employed on a small scale.

  17. Occurrence of Cryptosporidiumspp. oocysts in low quality water and on vegetables in Kumasi, Ghana

    DEFF Research Database (Denmark)

    Petersen, Tobias; Petersen, Heidi Huus; Abaidoo, Robert C.

    Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp. contaminat......Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp....... contamination level of river water, irrigation water and lettuce, 10L of water and 16 lettuce samples were collected four times in the period of, September – October 2013, with weekly intervals from six sample sites in and around Kumasi, Ghana. Oocysts were purified from water by sedimentation for 2 x 48 hours...... to characterize Cryptosporidium spp. positive samples were done by PCR amplification and sequencing of the SSU rRNA, the HSP70 and the GP60 genes after. Cryptosporidium oocysts were found in 75% of the water samples and on 43% of the lettuce with concentrations of 53 – 3268 per 10 L water and 11 – 118 oocyst per...

  18. Occurrence of Cryptosporidiumspp. oocysts in low quality water and on vegetables in Kumasi, Ghana

    DEFF Research Database (Denmark)

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

    2014-01-01

    Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp. contaminat......Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp....... contamination level of river water, irrigation water and lettuce, 10L of water and 16 lettuce samples were collected four times in the period of, September – October 2013, with weekly intervals from six sample sites in and around Kumasi, Ghana. Oocysts were purified from water by sedimentation for 2 x 48 hours...... to characterize Cryptosporidium spp. positive samples were done by PCR amplification and sequencing of the SSU rRNA, the HSP70 and the GP60 genes after. Cryptosporidium oocysts were found in 75% of the water samples and on 43% of the lettuce with concentrations of 53 – 3268 per 10 L water and 11 – 118 oocyst per...

  19. New insights into the excystation process and oocyst morphology of rodent Eimeria species.

    Science.gov (United States)

    Wiedmer, Stefanie; Stange, Joerg; Kurth, Thomas; Bleiss, Wilfrid; Entzeroth, Rolf; Kurth, Michael

    2011-10-01

    In this study, the mechanism of excystation of the rodent parasites Eimeria nieschulzi, from rats, and Eimeria falciformis, from mice, was investigated. In vitro, oocysts of both species are susceptible to the protease pepsin, and sporocysts and sporozoites can be excysted in a similar way. Scanning electron microscopy (SEM) revealed a collapse of the oocysts wall at both polar ends after pepsin treatment. This occurs without any visible damage of the outer wall. Using fluorescence and transmission electron microscopy (TEM) we observed that pepsin enters sporulated oocysts at both polar ends and causes degradation of the inner oocyst wall. Using scanning electron microscopy we could identify two polar caps in both investigated rodent Eimeria species, but only one is harbouring the micropyle. Thus the polar caps are the entry site for the pepsin. Furthermore, we provide evidence that the oocyst cap and micropyle are functionally different structures. This study complements the morphological description of both Eimeria species and is of relevance for other coccidian species.

  20. Uniform staining of Cyclospora oocysts in fecal smears by a modified safranin technique with microwave heating.

    Science.gov (United States)

    Visvesvara, G S; Moura, H; Kovacs-Nace, E; Wallace, S; Eberhard, M L

    1997-03-01

    Cyclospora, a coccidian protist, is increasingly being identified as an important, newly emerging parasite that causes diarrhea, flatulence, fatigue, and abdominal pain leading to weight loss in immunocompetent persons with or without a recent travel history as well as in patients with AIDS. Modified Kinyoun's acid-fast stain is the most commonly used stain to identify the oocyst of this parasite in fecal smears. Oocysts of Cyclospora stain variably by the modified acid-fast procedure, resulting in the possible misidentification of this parasite. We examined fecal smears stained by six different procedures that included Giemsa, trichrome, chromotrope, Gram-chromotrope, acid-fast, and safranin stains. We report on safranin-based stain that uniformly stains oocysts of Cyclospora a brilliant reddish orange, provided that the fecal smears are heated in a microwave oven prior to staining. This staining procedure, besides being superior to acid-fast staining, is fast, reliable, and easy to perform in most clinical laboratories.

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

    Science.gov (United States)

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

    2002-01-01

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

  2. Effect of subcutaneously administered diclazuril on the output of Eimeria species oocysts by experimentally infected rabbits.

    Science.gov (United States)

    Pan, B L; Zhang, Y F; Suo, X; Xue, Y

    2008-02-02

    The effect of subcutaneously injected diclazuril on the output of Eimeria species oocysts was studied in experimentally infected rabbits. Diclazuril was administered either prophylactically at 0.5, 1 or 2 mg/kg bodyweight two days before each rabbit was inoculated with 20,000 oocysts of a mixed-species field isolate of Eimeria or therapeutically at 1, 2 or 4 mg/kg bodyweight five days after they were inoculated. The prophylactic treatments significantly reduced (PEimeria stiedae, Eimeria magna, Eimeria irresidua, Eimeria flavescens, Eimeria piriformis and Eimeria intestinalis was significantly reduced (P<0.05) in all the treated groups. The burden of oocysts in the livers of the therapeutic groups (4000 to 9000) were significantly lower (P<0.05) than in the inoculated but untreated control group (23,000), but higher than in the prophylactic groups (around 1000).

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

    DEFF Research Database (Denmark)

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

    2016-01-01

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

  4. Experimental transmission of Cryptosporidium oocyst isolates from mammals, birds and reptiles to captive snakes.

    Science.gov (United States)

    Graczyk, T K; Cranfield, M R

    1998-01-01

    Groups of four to five, 3-month-old rat snakes (Elaphe obsoleta) were separately gastrically inoculated with 2.0 x 10(6) viable oocysts of Cryptosporidium muris (mice and calves), C. muris-like (Bactrian camels), C. wrairi (guinea pigs), C. baileyi (chickens), C. meleagridis (turkeys), Cryptosporidium sp. (turtles, tortoises, chameleons and lizards) and C. serpentis from clinically (fatal case) and subclinically infected snakes. None of the snakes inoculated with oocysts originating from homothermous vertebrates developed infection as determined by histology and serology, whereas all snakes challenged with reptilian oocyst isolates were infected with Cryptosporidium on weeks 6 and 10 post-inoculation (PI). One week 10 PI, the snakes displayed mild to serve, multifocal to widespread, thinning and disorganization of gastric epithelium and nine out of twelve snakes infected by oocysts originating from reptiles other than snakes displayed severe gastric hyperplasia. Three out of ten snakes infected oocysts originating from snakes had ELISA-detectable Cryptosporidium-specific antibody (Ab) titers on week 6 PI; all snakes were Cryptosporidium-seroconverted on week 10 PI and their serum Ab titer significantly increased. The study demonstrated that Cryptosporidium infections in snakes maintained on the diet of rodents or birds cannot be initiated via ingestion of an infected food item; however, snakes can void ingested oocysts. Lack of host specificity among reptiles to this pathogen, demonstrated for the first time in the present study, indicates that snake-attributed C. serpentis is not distinct from Cryptosporidium sp. infecting reptiles other than snakes, and that clinical manifestations and virulence of Cryptosporidium in snakes in modulated by the species of the host. Housing of snakes with other reptiles can enhance transmission of Cryptosporidium to snakes, and therefore should be avoided.

  5. Determination of the recovery efficiency of cryptosporidium oocysts and giardia cysts from seeded bivalve mollusks.

    Science.gov (United States)

    Schets, Franciska M; van den Berg, Harold H J L; de Roda Husman, Ana Maria

    2013-01-01

    The intestinal parasites Cryptosporidium and Giardia are transmitted by water and food and cause human gastroenteritis. Filter-feeding bivalve mollusks, such as oysters and mussels, filter large volumes of water and thus concentrate such pathogens, which makes these bivalves potential vectors of disease. To assess the risk of infection from consumption of contaminated bivalves, parasite numbers and parasite recovery data are required. A modified immunomagnetic separation (IMS) procedure was used to determine Cryptosporidium oocyst and Giardia cyst numbers in individually homogenized oysters (Crassostrea gigas) and mussels (Mytilus edulis). About 12% of the commercial bivalves were positive, with low (oo)cyst numbers per specimen. The recovery efficiency of the IMS procedure was systematically evaluated. Experiments included seeding of homogenized bivalves and whole animals with 100 to 1,000 (oo)cysts. Both seeding procedures yielded highly variable recovery rates. Median Cryptosporidium recoveries were 7.9 to 21% in oysters and 62% in mussels. Median Giardia recoveries were 10 to 25% in oysters and 110% in mussels. Giardia recovery was significantly higher than Cryptosporidium recovery. (Oo)cysts were less efficiently recovered from seeded whole animals than from seeded homogenates, with median Cryptosporidium recoveries of 5.3% in oysters and 45% in mussels and median Giardia recoveries of 4.0% in oysters and 82% in mussels. Both bivalve homogenate seeding and whole animal seeding yielded higher (oo)cyst recovery in mussels than in oysters, likely because of the presence of less shellfish tissue in IMS when analyzing the smaller mussels compared with the larger oysters, resulting in more efficient (oo)cyst extraction. The data generated in this study may be used in the quantitative assessment of the risk of infection with Cryptosporidium or Giardia associated with the consumption of raw bivalve mollusks. This information may be used for making risk management

  6. 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...

  7. Dispersion and Transport of Cryptosporidium Oocysts from Fecal Pats under Simulated Rainfall Events

    OpenAIRE

    Davies, Cheryl M.; Ferguson, Christobel M.; Kaucner, Christine; Krogh, Martin; Altavilla, Nanda; Deere, Daniel A.; Nicholas J. Ashbolt

    2004-01-01

    The dispersion and initial transport of Cryptosporidium oocysts from fecal pats were investigated during artificial rainfall events on intact soil blocks (1,500 by 900 by 300 mm). Rainfall events of 55 mm h−1 for 30 min and 25 mm h−1 for 180 min were applied to soil plots with artificial fecal pats seeded with approximately 107 oocysts. The soil plots were divided in two, with one side devoid of vegetation and the other left with natural vegetation cover. Each combination of event intensity a...

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

    Science.gov (United States)

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

    2013-09-01

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

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

    Science.gov (United States)

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

    1999-05-01

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

  10. 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 filte

  11. 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...

  12. 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.

  13. TOXOPLASMA GONDII : UPTAKE AND SURVIVAL OF OOCYSTS IN FREE-LIVING AMOEBAE

    Science.gov (United States)

    Waterborne transmission of the oocyst stage of Toxoplasma gondii can cause outbreaks of clinical toxoplasmosis in humans and infection of marine mammals. In water-related environments and soil, free-living amoebae are considered potential carriers of various pathogens, but knowledge on interactions ...

  14. Application of gel-bead technology for delivering Eimeria oocysts to day-old broilers

    Science.gov (United States)

    Current methods of preventing outbreaks of avian coccidiosis involve medication of feed with ionophore drugs or synthetic chemicals or by vaccination of chicks with low doses of Eimeria oocysts in ovo or by spray vaccination just after hatch. Our data indicates that the uniformity and efficiency of...

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

    Directory of Open Access Journals (Sweden)

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

    2000-01-01

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

  16. 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.

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

    Directory of Open Access Journals (Sweden)

    Srisuphanunt M.

    2009-09-01

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

  18. Herd factors influencing oocyst production of Eimeria and Cryptosporidium in Estonian dairy cattle.

    Science.gov (United States)

    Lassen, Brian; Viltrop, Arvo; Järvis, Toivo

    2009-10-01

    Cryptosporidium and Eimeria are intestinal parasites which are sensitive to the surroundings, behaviour and well-being of their host. In the present study, a range of factors related to farm management systems, environment, housing and herd characteristics were investigated with regard to alterations in oocyst excretion in cattle, using a mixed-effects model. Information and samples for three age categories were obtained from 45 Estonian dairy farms, located in 15 counties. Leaving the calf with the mother after birth reduced the risk of shedding higher levels of Cryptosporidium (OR = 0.20) and Eimeria (OR = 0.68) oocysts in all animals. The calves younger than 3 months kept on farms housing at least 150 animals had less risk (OR = 0.39) of producing higher numbers of Cryptosporidium oocysts. A somewhat lower infection level was observed in 3- to 12-month-old animals housed in separate buildings (OR = 0.64). The chance of shedding higher levels of Eimeria doubled (OR = 2.27) in cattle older than a year in case a vacancy period was used before replacing animals in pens and tripled (OR = 2.94) when the relative humidity exceeded 75% in the cowshed. Winter reduced the odds (OR = 0.25) of shedding Eimeria oocysts in the oldest animals compared to the fall season. Simple changes in handling and housing of cattle may produce a positive effect on controlling coccidian infections in Estonian dairy herds.

  19. 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

  20. 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 water samples were-positive). However, the number of cysts isolated per surface water sample was lower on average. Most water purification plants showed effective removal of cysts and oocysts. However, 13% of potable water samples contained protozoan...

  1. Soil contamination of Toxoplasma gondii oocysts in pig farms in central China.

    Science.gov (United States)

    Du, Fen; Zhang, Qingli; Yu, Qian; Hu, Min; Zhou, Yanqin; Zhao, Junlong

    2012-06-08

    Toxoplasmosis in pigs is a large threat to pig industry as well as pork consumers. Most pigs become infected by ingestion of oocysts from contaminated environment (soil, water and feed) or infected animal tissues postnatally. In the present study, field studies were conducted to evaluate the relationship between soil contamination status of Toxoplasma gondii oocysts and T. gondii infection in pigs in 12 pig farms with different density of cats in central China. The presence of T. gondii oocysts in soil were determined by PCR and loop-mediated isothermal amplification (LAMP). T. gondii DNA was found in 11 farms with different cat density excepting one farm exposed to low cat density. Twenty (21.1%) and 36 (37.9%) of 95 soil samples were T. gondii positive by PCR and LAMP, respectively (0.01Soil contamination was found with significant difference in statistics between farms with high and low cat density by PCR (0.01soil contamination by T. gondii oocysts in Hubei Province. The soil contamination status of pig farms was significantly influenced by cat density and may be an indicator of T. gondii infection in pigs.

  2. A gel delivery system for coccidiosis vaccine: uniformity of distribution of oocysts.

    Science.gov (United States)

    Dasgupta, T; Lee, E H

    2000-01-01

    A patented gel delivery system being used to deliver coccidiosis vaccine to poultry hatchlings is assessed. For effective vaccination, the coccidial oocysts must be uniformly suspended before exposure to birds. The uniformity of distribution within the gel was evaluated by incorporating a culture of chicken gut flora into gel sausages, placing sections of the sausage on culture plates, determining the appearance and distribution of bacterial colonies on culture plates after incubation, and verifying by cell counts. The uniformity of distribution of similarly prepared coccidial oocysts was verified by infecting birds with 40,000 Eimeria tenella oocysts delivered via the gel. Gel-inoculated birds were compared with control birds inoculated PO with 40,000 oocysts suspended in water by using cecal lesion scores. Both the appearance and colony counts of chicken gut flora from the gel were uniform. The standard deviation in the lesion scores for the gel-inoculated group and the water-inoculated groups were 0.51 and 0.69, respectively. The results indicate that a gel delivery system can provide uniform distribution of live organisms and vaccine agents to birds. Images Figure 1a. Figure 1b. PMID:10945126

  3. Aquatic biomonitoring of Giardia cysts and Cryptosporidium oocysts in peninsular Malaysia.

    Science.gov (United States)

    Lee, Soo Ching; Ngui, Romano; Tan, Tiong Kai; Roslan, Muhammad Aidil; Ithoi, Init; Lim, Yvonne A L

    2014-01-01

    An aquatic biomonitoring of Giardia cysts and Cryptosporidium oocysts in river water corresponding to five villages situated in three states in peninsular Malaysia was determined. There were 51.3% (20/39) and 23.1% (9/39) samples positive for Giardia and Cryptosporidium (oo)cysts, respectively. Overall mean concentration between villages for Giardia cysts ranged from 0.10 to 25.80 cysts/l whilst Cryptosporidium oocysts ranged from 0.10 to 0.90 oocysts/l. Detailed results of the river samples from five villages indicated that Kuala Pangsun 100% (9/9), Kemensah 77.8% (7/9), Pos Piah 33.3% (3/9) and Paya Lebar 33.3% (1/3) were contaminated with Giardia cysts whilst Cryptosporidium (oo)cysts were only detected in Kemensah (100 %; 9/9) and Kuala Pangsun (66.6%; 6/9). However, the water samples from Bentong were all negative for these waterborne parasites. Samples were collected from lower point, midpoint and upper point. Midpoint refers to the section of the river where the studied communities are highly populated. Meanwhile, the position of the lower point is at least 2 km southward of the midpoint and upper point is at least 2 km northward of the midpoint. The highest mean concentration for (oo)cysts was found at the lower points [3.15 ± 6.09 (oo)cysts/l], followed by midpoints [0.66 ± 1.10 (oo)cysts/l] and upper points [0.66 ± 0.92 (oo)cysts/l]. The mean concentration of Giardia cysts was highest at Kuala Pangsun (i.e. 5.97 ± 7.0 cysts/l), followed by Kemensah (0.83 ± 0.81 cysts/l), Pos Piah (0.20 ± 0.35 cysts/l) and Paya Lebar (0.10 ± 0.19 cysts/l). On the other hand, the mean concentration of Cryptosporidium oocysts was higher at Kemensah (0.31 ± 0.19 cysts/l) compared to Kuala Pangsun (0.03 ± 0.03cysts/l). All the physical and chemical parameters did not show significant correlation with both protozoa. In future, viability status and molecular characterisation of Giardia and Cryptosporidium should be applied to identify

  4. Transport and Retention of Toxoplasma gondii Oocysts in Loamy Sand and Sandy Loam Soils

    Science.gov (United States)

    Kinsey, E. N.; Korte, C.; L'Ollivier, C.; Dubey, J. P.; Aurélien, D.; Darnault, C. J. G.

    2016-12-01

    Toxoplasma gondii is one of the most prevalent parasites affecting warm-blooded animals and humans. It has a complex life cycle that involves a wide variety of intermediate hosts with felids as a definitive host. Humans may contract it through consumption of infected, undercooked meat or by water or food sources contaminated with the oocyst form of the parasite. Infection of pregnant women can cause stillbirth, neurological effects or blindness. Because of the prevalence of cats, including on farms where oocyst-contaminated cat feces, animal feed, soil and water have been found, T. gondii is spread almost throughout the entire globe. It has been implicated or suspected in waterborne infections since the 1990s. This study aims to characterize the transport and retention of T. gondii oocysts in field soils. The four soils used were collected from fallow and cultivated fields in Illinois and Utah, USA. They are classified as loamy sands and sandy loams. Soil columns were subjected to continuous artificial rainfall until they reached steady state at which point pulses that included 2.5 million T. gondii oocysts (Me49 strain) and KBr as a tracer were added. After the pulse infiltrated, continuous rainfall was resumed. Rain applied all columns was a 1 mM KCl solution. Leachate samples were collected, analyzed using qPCR for T. gondii and bromide ions and breakthrough curves were produced. Soil was sliced into 1 to 2 cm sections, for which water content and T. gondii concentration were measured to access degree of saturation and oocyst retention.

  5. 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.

  6. 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.

  7. A survey of Cryptosporidium oocysts in water supplies during a 10-year period (2000-2009) in Seoul.

    Science.gov (United States)

    Lee, Mok Young; Cho, Eun Joo; Lee, Jin Hyo; Han, Sun Hee; Park, Yong Sang

    2010-09-01

    This study has been conducted to estimate the occurrence of Cryptosporidium oocysts in water supplies in the Metropolitan area of Seoul, South Korea, for 10 years from 2000 to 2009. Water samples were collected quarterly at 6 intakes in the Han River and its largest stream and 6 conventional Water Treatment Plants (WTPs) serving drinking water for 10 million people of Seoul. Cryptosporidium oocysts were found in 22.5% of intake water samples and arithmetic mean was 0.65 oocysts/10 L (range 0-22 oocysts/10 L). Although the annual mean of oocyst number was as low as 0.04-1.90 oocysts/10 L, 3 peaks in 2004 and 2007 were observed and the pollution level was a little higher in winter. The lowest density was observed at Paldang intake and the pollution level increased at Kuui and Jayang intakes. At the end of the largest stream, oocysts were found in 70% of collected samples (mean 5.71 oocysts/10 L) and it seemed that its joining the Han River resulted in the increase at Kuui intake and downstream. Oocyst removal by physical process exceeded 2.0-2.3 log and then all finished water samples collected at 6 WTPs were negative for Cryptosporidium in each 100 L sample for 10 years. These results suggested that domestic wastewater from the urban region could be a source of Cryptosporidium pollution and separating sewage systems adjacent to the intakes could be meaningful for some intakes having weakness related to parasitological water quality.

  8. A slot blot immunoassay for quantitative detection of Plasmodium falciparum circumsporozoite protein in mosquito midgut oocyst.

    Directory of Open Access Journals (Sweden)

    Sanjai Kumar

    Full Text Available There is still a need for sensitive and reproducible immunoassays for quantitative detection of malarial antigens in preclinical and clinical phases of vaccine development and in epidemiology and surveillance studies, particularly in the vector host. Here we report the results of sensitivity and reproducibility studies for a research-grade, quantitative enhanced chemiluminescent-based slot blot assay (ECL-SB for detection of both recombinant Plasmodium falciparum circumsporozoite protein (rPfCSP and native PfCSP from Oocysts (Pf Oocyst developing in the midguts of Anopheles stephensi mosquitoes. The ECL-SB detects as little as 1.25 pg of rPfCSP (linear range of quantitation 2.5-20 pg; R2 = 0.9505. We also find the earliest detectable expression of native PfCSP in Pf Oocyst by ECL-SB occurs on day 7 post feeding with infected blood meal. The ECL-SB was able to detect approximately as few as 0.5 day 8 Pf Oocysts (linear quantitation range 1-4, R2 = 0.9795 and determined that one Pf Oocyst expressed approximately 2.0 pg (0.5-3 pg of native PfCSP, suggesting a similar range of detection for recombinant and native forms of Pf CSP. The ECL-SB is highly reproducible; the Coefficient of Variation (CV for inter-assay variability for rPf CSP and native PfCSP were 1.74% and 1.32%, respectively. The CVs for intra-assay variability performed on three days for rPf CSP were 2.41%, 0.82% and 2% and for native Pf CSP 1.52%, 0.57%, and 1.86%, respectively. In addition, the ECL-SB was comparable to microscopy in determining the P. falciparum prevalence in mosquito populations that distinctly contained either high and low midgut Pf Oocyst burden. In whole mosquito samples, estimations of positivity for P. falciparum in the high and low burden groups were 83.3% and 23.3% by ECL-SB and 85.7% and 27.6% by microscopy. Based on its performance characteristics, ECL-SB could be valuable in vaccine development and to measure the parasite prevalence in mosquitoes

  9. Neospora caninum-like oocysts observed in feces of free-ranging red foxes (Vulpes vulpes) and coyotes (Canis latrans).

    Science.gov (United States)

    Wapenaar, Wendela; Jenkins, Mark C; O'Handley, Ryan M; Barkema, Herman W

    2006-12-01

    The aim of this study was to examine the feces of free-ranging foxes and coyotes for the presence of Neospora caninum oocysts. Feces were collected from 271 foxes and 185 coyotes in the Canadian province of Prince Edward Island, processed by sucrose flotation, and examined by light microscopy for the presence of coccidian oocysts. In 2 fox and 2 coyote samples, oocysts morphologically and morphometrically similar to oocysts of N. caninum were observed. DNA was extracted from these samples and subjected to nested polymerase chain reaction (PCR) using primers to the N. caninum-specific Nc5 genomic sequence. Through DNA sequencing, alignment of the sequences of at least 3 clones from each isolate to sequences deposited in GenBank revealed 95-99% similarity to the Nc5 sequence of N. caninum. PCR using primers specific for Hammondia heydorni failed to yield an amplification product from these DNA samples.

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

    DEFF Research Database (Denmark)

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

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

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

    Science.gov (United States)

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

  12. Detection of Cryptosporidium oocysts and Giardia cysts in water samples with a Becton Dickinson FACSort flow cytometer

    NARCIS (Netherlands)

    Schets FM; Medema GJ; Boschman GD; LWL; Becton Dickinson Europe, Aalst, Belgie

    1995-01-01

    Current detection techniques for Cryptosporidium oocysts and Giardia cysts in water samples combine filtration of large volumes of water, concentration by centrifugation and flotation and immunofluorescense microscopy. The techniques are extremely labour-intensive and inefficient. The various steps

  13. A biocoagulant slow sand filtration for disinfection of Toxoplasma gondii oocysts from Mezam River in Bamenda, Cameroon

    National Research Council Canada - National Science Library

    Kenneth A Yongabi; Laura Knoll

    2015-01-01

    .... Dirty and turbid water (130.3 NTU) from Mezam River and leachates from dump sites and stagnant water in Bamenda, Cameroon, was analyzed microscopically after centrifugation for oocyst of T. gondii...

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

    Science.gov (United States)

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

    2013-11-01

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

  15. The development and performance of a simple, sensitive method for the detection of Cryptosporidium oocysts in faeces.

    OpenAIRE

    1984-01-01

    Features are described of a new staining method for the detection of Cryptosporidium oocysts in faeces. Smears, fixed in acid-methanol, are stained with heated safranin and counterstained with methylene blue. Oocysts stain a vivid orange-pink and are easily recognized. The method is rapid and simple with little source of error. The method is more sensitive than currently recommended Ziehl-Neelson methods; of 26 cases of cryptosporidiosis diagnosed by the new method only 19 were detected by ac...

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

    OpenAIRE

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

    1999-01-01

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

  17. Environmental Persistence and Infectivity of Oocysts of Two Species of Gregarines, Blabericola migrator and Blabericola cubensis (Apicomplexa: Eugregarinida: Blabericolidae), Parasitizing Blaberid Cockroaches (Dictyoptera: Blaberidae).

    Science.gov (United States)

    Clopton, Richard E; Steele, Shelby M; Clopton, Debra T

    2016-04-01

    For apicomplexan parasites using an oral-fecal transmission route with significant environmental exposure, the environmental persistence and infectivity of the oocyst has a direct impact on local infection dynamics, including the ability to withstand extended periods without readily available hosts. Herein we quantify the environmental persistence and infectivity of the oocysts of 2 septate gregarine species at controlled temperature and humidity and demonstrate that they can persist over multiple generational time spans. Species of Blabericola generally complete their endogenous life cycles from oocyst to oocyst within 10 days. The median residual environmental oocyst lifetime for Blabericola oocysts in this study is 21-28 days, but a significant number of oocysts of Blabericola migrator persisted and remained infective in the environment for up to 39 days while those of Blabericola cubensis persisted and remained infective for up to 92 days. Although long-lived relative to their own generational time, the oocysts of Blabericola species infecting cockroaches are short-lived relative to gregarines infecting tenebrionid beetles. For these gregarines, oocysts can persist in the environment and remain infective for up to 787 days. Mechanistically, environmental persistence and infectivity are probably energy-limited phenomena related to the amount of stored amylopectin and the basal metabolic rate of quiescent oocysts.

  18. Isolation of Cyclospora oocysts from fruits and vegetables using lectin-coated paramagnetic beads.

    Science.gov (United States)

    Robertson, L J; Gjerde, B; Campbell, A T

    2000-10-01

    Published techniques for recovering parasites from fruit and vegetables are generally inadequate, with low and variable recovery efficiencies. Herein, we describe an improved method for analyzing fruit and vegetables for Cyclospora oocysts. The technique includes washing procedures, sonication, and separation using lectin-coated paramagnetic beads. Identification is by microscopy (differential interference contrast and fluorescence). Oocyst recovery efficiencies from mushrooms, lettuce, and raspberries were approximately 12%. Recovery efficiencies from bean sprouts were approximately 4%. Although no significant difference in recovery efficiency could be detected between samples processed using the lectin-coated beads and samples processed without this procedure, distinct advantages were apparent when the lectin-coated beads were used. A considerably smaller, cleaner final volume remained for microscopy, which increases the sensitivity of the technique and reduces operator time.

  19. Anopheles gambiae eicosanoids modulate Plasmodium berghei survival from oocyst to salivary gland invasion

    Directory of Open Access Journals (Sweden)

    Susana Ramos

    2014-08-01

    Full Text Available Eicosanoids affect the immunity of several pathogen/insect models, but their role on the Anopheles gambiae response to Plasmodium is still unknown. Plasmodium berghei-infected mosquitoes were injected with an eicosanoid biosynthesis inhibitor, indomethacin (IN, or a substrate, arachidonic acid (AA, at day 7 or day 12 post-infection (p.i.. Salivary gland invasion was evaluated by sporozoite counts at day 21 p.i. IN promoted infection upon sporozoite release from oocysts, but inhibited infection when sporozoites were still maturing within the oocysts, as observed by a reduction in the number of sporozoites reaching the salivary glands. AA treatment had the opposite effect. We show for the first time that An. gambiae can modulate parasite survival through eicosanoids by exerting an antagonistic or agonistic effect on the parasite, depending on its stage of development.

  20. Anopheles gambiae eicosanoids modulate Plasmodium berghei survival from oocyst to salivary gland invasion.

    Science.gov (United States)

    Ramos, Susana; Custódio, Ana; Silveira, Henrique

    2014-08-01

    Eicosanoids affect the immunity of several pathogen/insect models, but their role on the Anopheles gambiae response to Plasmodium is still unknown. Plasmodium berghei-infected mosquitoes were injected with an eicosanoid biosynthesis inhibitor, indomethacin (IN), or a substrate, arachidonic acid (AA), at day 7 or day 12 post-infection (p.i.). Salivary gland invasion was evaluated by sporozoite counts at day 21 p.i. IN promoted infection upon sporozoite release from oocysts, but inhibited infection when sporozoites were still maturing within the oocysts, as observed by a reduction in the number of sporozoites reaching the salivary glands. AA treatment had the opposite effect. We show for the first time that An. gambiae can modulate parasite survival through eicosanoids by exerting an antagonistic or agonistic effect on the parasite, depending on its stage of development.

  1. Detection of oocysts of Cryptosporidium spp in gall bladder of sheep slaughtered in Mosul, Iraq

    Directory of Open Access Journals (Sweden)

    E. G. Suleiman

    2012-01-01

    Full Text Available The current study revealed the presence of oocysts of Cryptosporidium in the fluid and tissue of gall bladder of sheepwhich is slaughtered in Mosul city by using different diagnostic techniques, including direct smear method, staining method bymodified Zeil-Nelseen stain, scraping of mucosal layer of gall bladder and histological examination, the percentage ofinfection 20% and there were significant differences in the percentages of infection between gall bladder 20% and fecalsamples 41%. The histological study showed that the oocyst of Cryptosporidium was attached on the surface of the epithelialcells of gall bladder. The high percentage of infection appeared in ewes was 30% and there was no significant differences inthe infection of gall bladder between rams and ewes.

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

    Directory of Open Access Journals (Sweden)

    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.

  3. 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 sam....... auburnensis (p = 0.10). Further studies should focus on possible synergistic effects of multiple Eimeria spp. infections as well as interaction between Eimeria spp. and other enteric pathogens....

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

    OpenAIRE

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

    1990-01-01

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

  5. 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.

  6. First report of Cryptosporidium spp. oocysts in oysters (Crassostrea rhizophorae) and cockles (Tivela mactroides) in Brazil.

    Science.gov (United States)

    Guiguet Leal, Diego Averaldo; Pereira, Mirna Aparecida; Bueno Franco, Regina Maura; Branco, Nilson; Neto, Romeu Cantusio

    2008-12-01

    The consumption of oysters and cockles, which are usually eaten raw or lightly-cooked, can cause outbreaks of human diseases, especially if these shellfish are harvested from polluted areas. In Brazil data about the occurrence of pathogens, like hepatitis A virus, in shellfish have been reported but research on natural contamination for pathogenic protozoa is still non-existent. Cryptosporidium oocyst contamination of oysters (Crassostrea rhizophorae) and cockles (Tivela mactroides) was evaluated during two different periods in a coastal area from São Paulo, Brazil. From June to November 2005, and from July to December 2006, 180 mollusks were harvested for tissue examination. The gills and gastrointestinal tract (n = 36 pools) were carefully extracted from the animals and homogenized in a tissue homogenizer by adding surfactant Tween 80 (0.1%). Immunofluorescence assays were performed and Cryptosporidium oocysts were detected in 50.0% of gill pools of cockles and 10.0% of gill pools of oysters. In order to evaluate seawater quality in shellfish growing areas, total levels of thermotolerant coliforms, Escherichia coli and enterococci were determined. This is the first time that Cryptosporidium oocysts were found in shellfish from the coastal region of Brazil, and to the best of our knowledge it is also the first report in Latin America and the case might be of public health importance, reflecting the extension of the contamination on seafood, requiring a need for quality control standards. Copyright IWA Publishing 2008.

  7. 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.

  8. Effects of glass bead size, vortexing speed and duration on Eimeria acervulina oocyst excystation.

    Science.gov (United States)

    Cha, Jang-Ock; Talha, Abul Fatah Shah Muhammad; Lim, Chae Woong; Kim, Bumseok

    2014-03-01

    Improved methods for efficient excystation of Eimeria should be developed and standardized for future Eimeria-related studies. Here, the effects of different glass bead sizes (0.5, 1, 2, and 2.5 mm), and various vortex speeds (1000, 2000, and 3000 rpm) and durations (30 s, 1, 3, and 5 min) have been examined for Eimeria (E.) acervulina oocyst excystation. At 3000 rpm, all glass beads, regardless of size, efficiently ruptured E. acervulina oocysts at 5 min. At 2000 and 3000 rpm, all four glass bead sizes increasingly ruptured oocysts in a time-dependent manner. In contrast, at 1000 rpm the excystation efficiency was not related with the glass bead size or with vortexing duration. It appeared that the 1mm glass beads are most efficient for E. acervulina DNA extraction at a 3000 rpm vortexing speed for 3 and 5 min. The 2 mm glass beads delicately released the highest number of intact sporocysts at 2000 rpm for 3 min. Therefore, our data can provide valuable information for the efficient mechanical excystation of E. acervulina.

  9. Occurrence of Cryptosporidium spp. oocysts in the maintenance water of Antillean manatees (Trichechusmanatus in captivity

    Directory of Open Access Journals (Sweden)

    Jeane Kury Nobre Gomes

    2007-09-01

    Full Text Available The goal of this research was to verify the occurrence of Cryptosporidium spp. oocysts in the water consumption and supply of pools used by Antillean manatee (Trichechus manatus in captivity. Six samples were collected from the pool's supply system (at the beach, water used for manatee consumption, mineral water used in the preparation of artificial milk formulas offered to orphan manatee calves, also used in permanent maintenance pools in the visitation area, and water utilized in the rehabilitation area, where calves of all ages were kept. Before the water samples were processed, each sample was submitted to a filtration process. The diagnosis of the parasite was obtained by Kinyoun technique and the positive samples were submitted to the Direct Immunoflorescence Test. The results showed the presence of Cryptosporidium spp. oocysts in 66.67% (4/6 of the samples from the water supply system and pool, with eight to 56 oocysts per liter. There was evidence that the water used in the maintenance of the Antillean manatees could be an important medium for hydric transmission of Cryptosporidium spp. and that it may be a predisposing factor to the occurrence of cryptosporidiosis in Antillean manatees in captivity.

  10. Oocyst Discharge, Rumen Metabolism and Performance of Early Weaned Lambs with Naturally Occurring Coccidiosis Fed Monensin

    Science.gov (United States)

    Horton, G. M. J.; Stockdale, P. H. G.

    1981-01-01

    Ninety-six 9.5 kg early-weaned lambs with naturally occurring coccidiosis were fed monensin either at 0, 11, 22, or 33 mg/kg of feed for 105 days. Fecal oocyst discharge during the first three days was highest with monensin 22 mg, lowest with monensin 33 mg and averaged 149.6 × 103 oocysts per gram feces for all lambs. Monensin at 22 mg/kg of feed reduced Eimeria ninakohlyakimovae and E. ahsata oocyst discharge. Organic matter and crude protein digestibilities were highest (P ≤ 0.05) in lambs fed monensin 22 mg/kg of feed. Monensin increased (P ≤ 0.01) rumen ammonia and propionic acid and decreased (P ≤ 0.01) acetic acid. Feeding monensin 33 mg decreased (P ≤ 0.05) feed intake by 5% and had no effect on gain or feed efficiency. Optimal responses were observed with monensin at 11 mg, feed consumption was not affected, gains were 8% higher (P ≤ 0.05) and feed was utilized 9% more efficiently (P ≤ 0.05) than the controls. In conclusion, monensin was an effective therapeutic agent against naturally occurring coccidial infections in early weaned lambs. Performance responses were largest with monensin fed at the rate of 11 mg/kg of feed. PMID:7197187

  11. 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.

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

    Science.gov (United States)

    Bajszár, George; Dekonenko, Alexander

    2010-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Magali Roques

    2015-11-01

    Full Text Available 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.

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

    Science.gov (United States)

    Kothavade, Rajendra J

    2012-08-01

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

  15. DNA extraction from protozoan oocysts/cysts in feces for diagnostic PCR.

    Science.gov (United States)

    Hawash, Yousry

    2014-06-01

    PCR detection of intestinal protozoa is often restrained by a poor DNA recovery or by inhibitors present in feces. The need for an extraction protocol that can overcome these obstacles is therefore clear. QIAamp® DNA Stool Mini Kit (Qiagen) was evaluated for its ability to recover DNA from oocysts/cysts directly from feces. Twenty-five Giardia-positive, 15 Cryptosporidium-positive, 15 Entamoeba histolytica-positive, and 45 protozoa-free samples were processed as control by microscopy and immunoassay tests. DNA extracts were amplified using 3 sets of published primers. Following the manufacturer's protocol, the kit showed sensitivity and specificity of 100% towards Giardia and Entamoeba. However, for Cryptosporidium, the sensitivity and specificity were 60% (9/15) and 100%, respectively. A series of optimization experiments involving various steps of the kit's protocol were conducted using Cryptosporidium-positive samples. The best DNA recoveries were gained by raising the lysis temperature to the boiling point for 10 min and the incubation time of the InhibitEX tablet to 5 min. Also, using a pre-cooled ethanol for nucleic acid precipitation and small elution volume (50-100 µl) were valuable. The sensitivity of the amended protocol to Cryptosporidium was raised to 100%. Cryptosporidium DNA was successfully amplified by either the first or the second primer set. When applied on parasite-free feces spiked with variable oocysts/cysts counts, ≈ 2 oocysts/cysts were theoretically enough for detection by PCR. To conclude, the Qiagen kit with the amended protocol was proved to be suitable for protozoan DNA extraction directly from feces and support PCR diagnosis.

  16. 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.

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

    Science.gov (United States)

    Lalancette, Cindy; Di Giovanni, George D.; Prévost, Michèle

    2010-01-01

    The inactivation of Cryptosporidium oocysts is a main driver in the selection of water treatment disinfection strategies, and microbial risk analysis provides a sound basis for optimizing water treatment processes. U.S. Environmental Protection Agency method 1622/23 provides an estimate of the total oocyst count; however, it cannot be used directly for risk assessment, as it does not determine the fraction of infectious oocysts. Improved assessment of the risk for designated sources or in treated water requires evaluation of the total number of oocysts and an estimate of their infectivity. We developed a dual direct detection method using differential immunofluorescent staining that allows detection of both oocysts and cell culture infection foci for each sample. Using Cryptosporidium parvum oocysts, various pH levels, proteases, and gastroenteric compounds and substrates were assessed to determine their abilities to enhance the number of infection foci. The results showed that the key trigger for oocyst stimulation was acidification. Addition of a low concentration of d-glucose (50 mM) to the infection media increased rates of infectivity, while a higher dose (300 mM) was inhibitory. The total number of oocysts in each sample was determined by counting the oocysts remaining on a cell monolayer and the oocysts recovered from cell monolayer washes during processing using a simple filtration technique. With the dual direct detection on cell culture with immunofluorescence assay method, it is now possible to determine the numbers of total and infectious oocysts for a given sample in a single analysis. Direct percentages of infectivity are then calculated, which allows more accurate assessments of risk. PMID:19933339

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

    DEFF Research Database (Denmark)

    Sommer, C.

    1998-01-01

    This study reports on morphological variability of Eimeria species, which may be given either by drawings or as quantitative data. The drawings may be used to facilitate identification by eye of 'unknown' Eimeria specimens, whereas quantitative data may serve as a reference set for identification...... 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...

  19. 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

    Background: Sonication of fluids has been widely used for cell lyses and decontamination of water and food due to its three major physical and chemical effects: cavitation, implosion of water bubbles with subsequent increase in substrate temperature and liberation of free radicals such as O....... Results: The results showed that exposure of Cryptosporidium oocysts and Giardia cysts to ultrasound could reduce their survival rate even if the exposure time was limited to a few seconds. When sonication time was extended, changes in parasite characteristics became clearly visible. Several other factors...

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

    DEFF Research Database (Denmark)

    Petersen, Tobias Boel; Petersen, Heidi Huus; Robert C., Abaidoo

    Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp....... contamination level of river water, irrigation water and lettuce, 10L of water and 16 lettuce samples were collected four times in the period September – October 2013, with weekly intervals from six sample sites in and around Kumasi, Ghana. Oocysts were purified from water by sedimentation for 2 x 48 hours...... off the slides and attempts to characterize Cryptosporidium spp. positive samples were done by PCR amplification and sequencing of the SSU rRNA, the HSP70 and the GP60 genes. Cryptosporidium oocysts were found in 75% of the water samples and on 43% of the lettuce with concentrations of 53 – 3,268 per...

  1. Detection of Cryptosporidium spp. Oocysts in raw sewage and creek water in the city of São Paulo, Brazil

    Directory of Open Access Journals (Sweden)

    Farias Eveline Wilma Coutinho

    2002-01-01

    Full Text Available The protozoan parasite Cryptosporidium has emerged as one of the most important contaminants of water, causing waterborne outbreaks of gastroenteritis worldwide. To monitor and understand the public health significance of this pathogen in environmental samples, several methods have been developed to isolate and detect Cryptosporidium oocysts. The purpose of this study was to perform the first investigation on the presence of Cryptosporidium spp. oocysts in raw sewage and creek water in the city of São Paulo, Brazil. The oocysts were concentrated by flocculation and membrane filtration. The results showed the occurrence of Cryptosporidium spp. in all wastewater samples analyzed, indicating a possible risk for dissemination of these pathogens in aquatic environment and in the community.

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

    Science.gov (United States)

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

    2007-01-01

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

  3. Beta-1,3-glucan1,3-glucan, a druggable target, forms a trabecular scaffold in the oocyst walls of Toxoplasma and Eimeria

    Science.gov (United States)

    Toxoplasma gondii is a human pathogen that spreads by ingestion of oocysts in cat feces or tissue cysts in raw meat. Eimeria species are major pathogens of livestock that also spread by infectious oocysts. Beta-1,3-glucan, a major component of fungal and plant walls, is the target of anti-fungal dr...

  4. Development and evaluation of an off-the-slide genotyping technique for identifying Giardia cysts and Cryptosporidium oocysts directly from US EPA Method 1623 slides

    Science.gov (United States)

    ABSTRACT Aims This study developed and systematically evaluated performance and limit of detection of an off-the-slide genotyping procedure for both Cryptosporidium oocysts and Giardia cysts. Methods and Results Slide standards containing flow sorted (oo)cysts were used to e...

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

    Directory of Open Access Journals (Sweden)

    Zhang Haili

    2012-11-01

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

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

    Science.gov (United States)

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

    2010-06-01

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

  7. Effects of different sizes of glass beads on the release of sporocysts from Eimeria tenella oocysts.

    Science.gov (United States)

    You, Myung-Jo

    2014-06-01

    The oocyst wall is severed by means of mechanical injury or chemical agents. This study reports the percentage of in vitro sporocyst release following mechanical shaking in the presence of varying sizes of glass beads. Glass beads measured 0.5, 1, and 3 mm in diameter and were shaken with the oocysts for different times ranging from 5 sec to 5 min. Approximately 80% of sporocysts were released with 5 min of shaking in the presence of 3 mm glass beads, as well as 30 sec with 0.5 mm beads and 1 mm glass beads. The release of sporocysts of E. tenella was most efficient using 1 mm glass beads and treatment times of 30 sec to 1 min. Therefore, the use of 1 mm glass beads with 30 sec to 1 min of agitation is recommended in order to maximize sporocyst release and recovery and to improve the yield of viable sporozoites for use in biochemical, tissue culture, and immunological applications of coccidia.

  8. 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...

  9. INACTIVATION OF CRYPTOSPORIDIUM OOCYSTS IN A PILOT-SCALE OZONE BUBBLE-DIFFUSER CONTACTOR - II: MODEL VALIDATION AND APPLICATION

    Science.gov (United States)

    The ADR model developed in Part I of this study was successfully validated with experimenta data obtained for the inactivation of C. parvum and C. muris oocysts with a pilot-scale ozone-bubble diffuser contactor operated with treated Ohio River water. Kinetic parameters, required...

  10. 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 Ass

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

    DEFF Research Database (Denmark)

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

    Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp. contaminat......Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp....... contamination level of river water, irrigation water and lettuce, 10L of water and 16 lettuce samples were collected four times in the period September – October 2013, with weekly intervals from six sample sites in and around Kumasi, Ghana. Oocysts were purified from water by sedimentation for 2 x 48 hours...... off the slides and attempts to characterize Cryptosporidium spp. positive samples were done by PCR amplification and sequencing of the SSU rRNA, the HSP70 and the GP60 genes. Cryptosporidium oocysts were found in 75% of the water samples and on 43% of the lettuce with concentrations of 53 – 3,268 per...

  12. 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 Ass

  13. 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...

  14. Analytical sensitivity of staining and molecular techniques for the detection of Cryptosporidium spp. oocysts isolated from bovines in water samples: a preliminary study

    Directory of Open Access Journals (Sweden)

    A Díaz-Lee

    2015-01-01

    Full Text Available Cryptosporidium spp. is a globally distributed protozoan that causes digestive disease in different animals including humans. Excreted oocysts contaminate water and soil, constituting a public health threat. Sensitive and fast methods to detect oocysts in water samples are necessary due to the small number of oocysts present in the environment and their low infectious dose. This study compared the analytical sensitivity of two staining techniques, modified Ziehl-Neelsen and Auramine versus a nested PCR that amplifies a region of ~520bp from 18S rDNA gene, to detect Cryptosporidium spp. in water samples. Water was inoculated with oocysts using serial dilutions, and then a water filtration method was used to recover the parasite oocysts. The staining techniques had similar analytical sensitivity, detecting 8 oocysts/mL, while the nested PCR detected down to 6 oocysts/mL. In conclusion, all of these methods are effective for Cryptosporidium spp. detection in water samples, contributing to the implementation of standardized diagnostic methods for environmental water infectious agents.

  15. 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.

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

    Science.gov (United States)

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

    1990-09-01

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

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

    DEFF Research Database (Denmark)

    Enemark, Heidi

    2015-01-01

    -time sensor detection of Cryptosporidium and Giardia in drinking water. This novel optical technique, in combination with advanced data analysis, yields a measure for the protozoal content present in a sample. High sensitivity of the system is acquired through a combination of a new, patented filtration......To secure the safety and quality of food and water presence of pathogens including parasites must be controlled. This relies among other things on implementation of sensitive and specific tests which are able to rule out the occurrence of parasites. Worldwide environmentally resistant oocysts...... use of a metallic filter, sonication and "air backwash" are key factors in the creation of this highly efficient and robust system which can be used continuously for extended time with minimal maintenance requirements. Sample acquisition and analysis is performed in real-time where objects...

  18. Isolation and enumeration of Giardia cysts, cryptosporidium oocysts, and Ascaris eggs from fruits and vegetables.

    Science.gov (United States)

    Robertson, L J; Gjerde, B

    2000-06-01

    Published techniques for recovering parasites from fruit and vegetables are generally inadequate, with low and variable recovery efficiencies. Here we describe an improved methodology for analyzing fruit and vegetables for Giardia cysts, Cryptosporidium oocysts, and Ascaris eggs. The method includes washing procedures, sonication, and, for Giardia and Cryptosporidium, immunomagnetic separation. Identification is by immunofluorescence (Giardia and Cryptosporidium) or brightfield microscopy (Ascaris). Recovery efficiencies from lettuce, Chinese leaves, and strawberries were found to be approximately 67% for Giardia, 42% for Cryptosporidium, and 72% for Ascaris. Recovery efficiencies from bean sprouts tended to be more variable and lower. This could be due to material removed with the parasites during the washing procedures, which, in turn, appeared related to the age of the bean sprouts. It is therefore recommended that fruit and vegetables should be as fresh as possible when analyzed for parasites.

  19. 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

    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...... at predetermined time intervals to loosen all parasites and other particles from the filter and break up clusters. The concentrated parasite suspension was backwashed and counted after immuno-fluorescence staining. Results and discussion: Without sonication the recovery rates of both Cryptosporidium and Giardia...... chamber further facilitated the concentration of parasites by ensuring a backwash volume of less than 1ml. The presented design of the filter system can be used as a cheap and time effective method of isolating water borne parasites in the laboratory....

  20. Hydrological modelling of Toxoplasma gondii oocysts transport to investigate contaminated snowmelt runoff as a potential source of infection for marine mammals in the Canadian Arctic.

    Science.gov (United States)

    Simon, Audrey; Rousseau, Alain N; Savary, Stéphane; Bigras-Poulin, Michel; Ogden, Nicholas H

    2013-09-30

    Toxoplasma gondii (T. gondii) is a zoonotic protozoan that sometimes causes serious illness in humans and other animals worldwide, including the Canadian Arctic. Wild and domestic felids, the only hosts able to shed T. gondii oocysts, are practically non-existent in the Canadian Arctic. So here the hypothesis that T. gondii oocysts, shed in the southern areas of the boreal watershed, could contaminate the Arctic coastal marine environment via surface runoff, particularly during the spring snowmelt period, was explored. A watershed model was applied to simulate the hydrological transport of T. gondii oocysts during the snowmelt period and test the possible efficiency of river-to-sea transport as a potential source of marine organisms' exposure to this pathogen. Simulations were run for two pilot watersheds with the ultimate aim of extrapolating the results across the Canadian Arctic watersheds. Results suggest that daily stream flow concentrations of T. gondii oocysts at the river outlet are likely to be very low. However, accumulation of oocysts in the estuarine areas may be large enough to contaminate estuarine/marine filter-feeding molluscs and snails on which seals and other marine mammals may feed. Potential maximum concentrations of T. gondii oocysts in runoff are reached at the beginning of the snowmelt period with maxima varying with discharge rates into rivers and how far upstream oocysts are discharged. Meteorological conditions during the snowmelt period can affect simulated concentrations of oocysts. These findings support the hypothesis that T. gondii oocysts carried in snowmelt runoff could be a source of T. gondii infection for marine mammals in the Canadian Arctic, and for Arctic human populations that hunt and consume raw meat from marine mammals.

  1. Eimeria involved in a case of coccidiosis in farmed red-legged partridges (Alectoris rufa) in France: oocyst isolation and gross lesion description after experimental infection.

    Science.gov (United States)

    Naciri, M; Repérant, J M; Fort, G; Crespin, J; Duperray, J; Benzoni, G

    2011-10-01

    The aim of the present work was, after a coccidiosis outbreak in a farm rearing red-legged partridges (Alectoris rufa) in Brittany (France), to identify the Eimeria species and describe gross lesions induced by three of them (Eimeria kofoidi, Eimeria caucasica and Eimeria legionensis) after experimental infection. E. kofoidi oocysts measured 19.3 µm × 16.3 µm on average; neither micropyle nor oocyst residuum were present, but one, two or more small polar granules were visible. After inoculation of 300,000 oocysts per partridge, severe gross lesions were observed in the duodenum and jejunum, characterized by thickened oedematous mucosa and lumen filled with thick mucus, gas and sometimes false-membrane due to sloughed epithelium. E. caucasica oocysts were on average 29.8 µm × 19.5 µm in size; no oocyst residuum was observed, but a large granule was well visible. E. caucasica also invaded both the duodenum and jejunum, causing haemorrhagic points on the serosal surface, as well as mucoid duodenitis and catarrhal enteritis when 30,000 oocysts were inoculated per bird. E. legionensis oocysts measured 22.6 µm × 14.9 µm on average; they presented a clear micropyle beneath which one or two granulations were present. E. legionensis mainly invaded the caeca; low mortality was observed at the dosage of 200,000 oocysts per bird. Caecal walls were thickened and caseous material was condensed into off-white cheesy cores. For each species, oocyst shedding started 5 days post inoculation, peaked at 9, 8 and 6 days post inoculation for E. kofoidi, E. caucasica and E. legionensis, respectively, then decreased and persisted until 15 days post inoculation (end of examinations).

  2. Seasonality in the proportions of domestic cats shedding Toxoplasma gondii or Hammondia hammondi oocysts is associated with climatic factors.

    Science.gov (United States)

    Schares, G; Ziller, M; Herrmann, D C; Globokar, M V; Pantchev, N; Conraths, F J

    2016-04-01

    A previous study on domestic cats in Germany and neighbouring countries suggested seasonality in shedding Toxoplasma gondii oocysts. The aim of the present study was to elucidate whether this seasonality in shedding could be explained by climatic effects and whether differences between years in the proportions of cats shedding oocysts could also be explained by climatic factors. To this end, a long-term study over a period of 55 months on domestic cats for T. gondii and Hammondia hammondi oocysts was performed and the results compared with climatic data. Using species-specific PCR, T. gondii oocysts were identified in 0.14% (84/61,224) and H. hammondi in 0.10% (61/61,224) of the samples. Toxoplasma gondii oocysts were predominantly observed from summer to autumn, while H. hammondi oocysts were mainly found during autumn and winter. In statistical analyses using climatic data, even differences in parasitological findings between years could be partially modelled using monthly temperature, North Atlantic Oscillation indices and precipitation. Of the three climatic variables analysed, precipitation as an explanatory variable had the lowest impact in the statistical models while those taking only temperature and North Atlantic Oscillation indices into account were sufficiently predictive. Interestingly, time lags between the climatic event and the parasitological findings had to be implemented in all models. For T. gondii, North Atlantic Oscillation indices with a time lag of 7 months and temperature with a time lag of 2 months had the best predictive value. In contrast, temperature (with a time lag of 6 months) and the interaction of precipitation (with a time lag of 5 months) and North Atlantic Oscillation indices (with a time lag of 11 months) were optimal for predicting the seasonality of H. hammondi. These results suggest prominent differences in the life cycles of the two closely related parasites. Previous findings showed that H. hammondi lack avian hosts, in

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

    Science.gov (United States)

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

    2016-10-01

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

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

    DEFF Research Database (Denmark)

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

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

  5. Oocyst shedding by green-winged-saltator (Saltator similis) in the diagnostic of coccidiosis and Isospora similisi n. sp. (Apicomplexa: Eimeriidae)

    National Research Council Canada - National Science Library

    Coelho, Cleide Domingues; Berto, Bruno Pereira; Neves, Daniel Medeiros; Oliveira, Vinícius Modesto de; Flausino, Walter; Lopes, Carlos Wilson Gomes

    2013-01-01

    .... The objective of this study was to evaluate the occurrence of periodicity of oocysts presence in the green-winged-saltator Saltator similis, and its use in the diagnosis of coccidiosis in wild birds in captivity...

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

    Science.gov (United States)

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

    2011-06-30

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

  7. 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.

  8. Recovery of Cryptosporidium oocysts from small and large volume water samples using a compressed foam filter system.

    Science.gov (United States)

    Sartory, D P; Parton, A; Parton, A C; Roberts, J; Bergmann, K

    1998-12-01

    A novel filter system comprising open cell reticulated foam rings compressed between retaining plates and fitted into a filtration housing was evaluated for the recovery of oocysts of Cryptosporidium from water. Mean recoveries of 90.2% from seeded small and large volume (100-2000 l) tap water samples, and 88.8% from 10-20 l river water samples, were achieved. Following a simple potassium citrate flotation concentrate clean-up procedure, mean recoveries were 56.7% for the tap water samples and 60.9% for river water samples. This represents a marked improvement in capture and recovery of Cryptosporidium oocysts from water compared with conventional polypropylene wound cartridge filters and membrane filters.

  9. The prevalence of Cryptosporidium spp. oocysts in wild mammals in the Kruger National Park, South Africa.

    Science.gov (United States)

    Abu Samra, Nada; Jori, Ferran; Samie, Amidou; Thompson, Peter

    2011-01-10

    This study determined the prevalence of Cryptosporidium spp. oocysts in faecal samples from elephant (Loxodonta africana), buffalo (Syncerus caffer) and impala (Aepyceros melampus) in the Kruger National Park (KNP) and an adjacent game reserve in South Africa. Two of the study areas were in close proximity to rural communities on the western KNP boundary and the third study area was located in the centre of the KNP. Fresh stool samples (n=445) were collected and tested using an immunofluorescent antibody test (IFA) for Cryptosporidium parvum. A total of 278 of these were randomly selected (approximately 90 samples per wildlife species) and tested with the modified Ziehl Neelsen staining technique (ZN) for Cryptosporidium spp. The prevalence of Cryptosporidium spp. was highest in elephants (25.8% [95% confidence interval: 17.3, 35.9]), compared to buffalo (5.5% [1.8, 12.4]) and impala (4.3% [1.2, 10.5]). C. parvum showed similar patterns, being most prevalent in elephants (4.2% [1.5, 8.8]), compared to buffalo (1.4% [0.2, 5.1]) and impala (1.9% [0.4, 5.3]). 29 samples, including ZN positive and IFA positive samples, were retested using a real time PCR (rtPCR) technique. Of the 28 ZN-positive samples, 14 (50%) were positive with rtPCR and of the 9 IFA-positive samples 6 (67%) were confirmed positive by rtPCR. The prevalence of Cryptosporidium oocysts was significantly higher in both of the two study areas adjacent to the western KNP boundary compared to the area in the centre of the KNP (OR=3.2 [1.2, 9.0]; P=0.024). Our study demonstrates for the first time the presence of Cryptosporidium spp. in wildlife in South Africa. The transmission of this parasite between wildlife, domestic animals and humans is a plausible hypothesis and represents a potential risk for immunodeficient human populations.

  10. Development of resistance to coccidiosis in the absence of merogonic development using X-irradiated Eimeria acervulina oocysts

    Energy Technology Data Exchange (ETDEWEB)

    Jenkins, M.C.; Augustine, P.C.; Barta, J.R.; Castle, M.D.; Danforth, H.D. (US Dept. of Agriculture, Beltsville, MD (USA))

    1991-04-01

    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.

  11. 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.

  12. Prevalence and genotypes of Toxoplasma gondii in feline faeces (oocysts) and meat from sheep, cattle and pigs in Switzerland.

    Science.gov (United States)

    Berger-Schoch, A E; Herrmann, D C; Schares, G; Müller, N; Bernet, D; Gottstein, B; Frey, C F

    2011-05-11

    The protozoan parasite Toxoplasma gondii infects almost all warm blooded animal species including humans, and is one of the most prevalent zoonotic parasites worldwide. Post-natal infection in humans is acquired through oral uptake of sporulated T. gondii oocysts or by ingestion of parasite tissue cysts upon consumption of raw or undercooked meat. This study was undertaken to determine the prevalence of oocyst-shedding by cats and to assess the level of infection with T. gondii in meat-producing animals in Switzerland via detection of genomic DNA (gDNA) in muscle samples. In total, 252 cats (44 stray cats, 171 pet cats, 37 cats with gastrointestinal disorders) were analysed coproscopically, and subsequently species-specific identification of T. gondii oocysts was achieved by Polymerase Chain Reaction (PCR). Furthermore, diaphragm samples of 270 domestic pigs (120 adults, 50 finishing, and 100 free-range animals), 150 wild boar, 250 sheep (150 adults and 100 lambs) and 406 cattle (47 calves, 129 heifers, 100 bulls, and 130 adult cows) were investigated by T. gondii-specific real-time PCR. For the first time in Switzerland, PCR-positive samples were subsequently genotyped using nine PCR-restriction fragment length polymorphism (PCR-RFLP) loci (SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico) for analysis. Only one of the cats shed T. gondii oocysts, corresponding to a T. gondii prevalence of 0.4% (95% CI: 0.0-2.2%). In meat-producing animals, gDNA prevalence was lowest in wild boar (0.7%; 95% CI: 0.0-3.7%), followed by sheep (2.0%; 95% CI: 0.1-4.6%) and pigs (2.2%; 95% CI: 0.8-4.8%). The highest prevalence was found in cattle (4.7%; 95% CI: 2.8-7.2%), mainly due to the high prevalence of 29.8% in young calves. With regard to housing conditions, conventional fattening pigs and free-range pigs surprisingly exhibited the same prevalence (2.0%; 95% CI: 0.2-7.0%). Genotyping of oocysts shed by the cat showed T. gondii with clonal Type II alleles and the Apico I

  13. Description of the Oocysts of Three New Species of Eimeria (Apicomplexa: Eimeriidae from Iguanid Lizards (Sauria: Iguanidae of Central and South America

    Directory of Open Access Journals (Sweden)

    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.

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

    Institute of Scientific and Technical Information of China (English)

    陈甫

    2011-01-01

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

  15. Effects of Developmental Temperature on Gametocysts and Oocysts of Two Species of Gregarines Blabericola migrator and Blabericola cubensis (Apicomplexa: Eugregarinida: Blabericolidae) Parasitizing Blaberid Cockroaches (Dictyoptera: Blaberidae).

    Science.gov (United States)

    Kolman, Jon A; Clopton, Richard E; Clopton, Debra T

    2015-12-01

    Abiotic environmental conditions, especially temperature and humidity, have profound effects on the growth and development of gregarines, but these effects remain largely undocumented. Quantifying the effects of environmental conditions on the growth and development of exogenous gregarine ontogenetic stages is an important first step in understanding the transmission, population dynamics, and environmental persistence of gregarine infection. In this study, we examined the effect of 6 environmental temperatures (10, 18, 22, 27, 35, and 40 C) at constant humidity (0 mmHg vapor pressure deficit) on gametocyst development and oocyst viability in 2 gregarine species: Blabericola migrator and Blabericola cubensis parasitizing the Tiger-striped Hissing Cockroach, Princisia vanwaerebecki, and the Discoid Cockroach, Blaberus discoidalis, respectively. Temperature has a significant effect on gametocyst development and oocyst viability for both gregarine species. Gametocyst development for both gregarine species displays a similar threshold response to environmental temperature: 10 and 40 C represent extremes outside their developmental range, but within these extremes, the relationship between gametocyst development and temperature is weakly direct. Dehiscence increased with temperature from 68% at 18 C to 93% at 22 C and remained at that level through 35 C. Developmental temperature also has a meaningful but inverse effect on oocyst viability of both B. migrator and B. cubensis. For both species, oocyst viability is highest at 18 and 22 C and is significantly reduced at 27 and 35 C. Thus oocyst production and sporozoite viability are linked but environmentally independent phenomena. Overall, there is an acceptable developmental temperature zone for B. migrator and B. cubensis that ranges from 18 to 27 C, but production of viable sporozoites is greatest in a relatively narrow zone around 22 C. Prior studies have postulated that mechanisms that concentrate oocysts and hosts

  16. Survey on the contamination of Toxoplasma gondii oocysts in the soil of public parks of Wuhan, China.

    Science.gov (United States)

    Du, F; Feng, H L; Nie, H; Tu, P; Zhang, Q L; Hu, M; Zhou, Y Q; Zhao, J L

    2012-03-23

    Toxoplasma gondii of warm-blooded animals and humans is an important pathogenic agent throughout the world. Soil is increasingly recognized as an important source in the transmission of Toxoplasma. To attain the contamination status of T. gondii in the soil of public parks, a total of 252 soil samples were collected from September 2009 to August 2010 at different sites located in 6 public parks of Wuhan, Hubei, China and detected by PCR and loop-mediated isothermal amplification (LAMP). The detection limit of PCR/B1, PCR/529 and LAMP was determined to be 50, 5, and 5 tachyzoites in soil, respectively. Forty-one samples were found positive for Toxoplasma DNA by PCR on both genes, whereas LAMP products were generated in 58 samples (χ(2)=3.6328, P=0.0567). All parks were found contaminated and no significant difference was found among the parks (PCR: χ(2)=0.0072, P=0.9325; LAMP: χ(2)=0.6101, P=0.4347). However, contamination was found with significantly different among the four seasons (PCR: χ(2)=11.6066, P=0.0007; LAMP: χ(2)=12.4636, P=0.0004), with a gradual decrease in the prevalence from spring to winter on both analyses. This is the first investigation on soil contamination of public parks in China by T. gondii oocysts. The results indicate that the soil of public parks contaminated with T. gondii oocysts may play a role in the epidemiology of toxoplasmosis and effective preventive measures should be considered. Moreover, the conventional PCR and LAMP used in the present study are applicable to detect T. gondii oocysts in soil samples.

  17. Oocyst-Derived Extract of Toxoplasma Gondii Serves as Potent Immunomodulator in a Mouse Model of Birch Pollen Allergy.

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    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.

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

    Science.gov (United States)

    Kunitomo, Shinta

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

  19. Roles of tyrosine-rich precursor glycoproteins and dityrosine- and 3,4-dihydroxyphenylalanine-mediated protein cross-linking in development of the oocyst wall in the coccidian parasite Eimeria maxima

    DEFF Research Database (Denmark)

    Belli, Sabina I; Wallach, Michael G; Luxford, Catherine

    2003-01-01

    (macrogamete) of Eimeria maxima are proteolytically processed into smaller glycoproteins, which are then incorporated into the developing oocyst wall. The identification of high concentrations of dityrosine and 3,4-dihydroxyphenylalanine (DOPA) in oocyst extracts by high-pressure liquid chromatography...... cuticles, yeast cell walls, mussel byssal threads, and sea urchin fertilization membranes....

  20. Elevation and vegetation determine Cryptosporidium oocyst shedding by yellow-bellied marmots (Marmota flaviventris in the Sierra Nevada Mountains

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    Diego Montecino-Latorre

    2015-08-01

    Full Text Available Wildlife are increasingly recognized as important biological reservoirs of zoonotic species of Cryptosporidium that might contaminate water and cause human exposure to this protozoal parasite. The habitat range of the yellow-bellied marmot (Marmota flaviventris overlaps extensively with the watershed boundaries of municipal water supplies for California communities along the foothills of the Sierra Nevada. We conducted a cross-sectional epidemiological study to estimate the fecal shedding of Cryptosporidium oocysts by yellow-bellied marmots and to quantify the environmental loading rate and determine risk factors for Cryptosporidium fecal shedding in this montane wildlife species. The observed proportion of Cryptosporidium positive fecal samples was 14.7% (33/224, positive number relative to total number samples and the environmental loading rate was estimated to be 10,693 oocysts animal-1 day-1. Fecal shedding was associated with the elevation and vegetation status of their habitat. Based on a portion of the 18s rRNA gene sequence of 2 isolates, the Cryptosporidium found in Marmota flaviventris were 99.88%–100% match to multiple isolates of C. parvum in the GenBank.

  1. 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...

  2. Extra-intestinal localization of Goussia sp. (Apicomplexa) oocysts in Rana dalmatina (Anura: Ranidae), and the fate of infection after metamorphosis.

    Science.gov (United States)

    Jirků, M; Modrý, D

    2006-06-23

    Although coccidia of the genus Goussia are common parasites of fish, only 2 species have been described in amphibians: G. hyperolisi from common reed frogs Hyperolius viridiflavus from Kenya and G. neglecta from unspecified European water frogs of the genus Rana from Germany. The genus Goussia is characterized by an oocyst, with a fine oocyst wall, containing 4 dizoic sporocysts that are composed of 2 valves joined by a longitudinal suture and lacking a Stieda body (typical for the genus Eimeria). To date, infections in amphibians were generally considered to be specific to the intestine of aquatic larval stages (tadpoles) of anurans. Herein, we report on: (1) the presence of oocysts of Goussia sp. in an extra-intestinal location (liver) of tadpoles of the agile frog R. dalmatina and (2) the presence of oocysts in the liver of both juvenile and subadult R. dalmatina. These observations represent novel traits for Goussia infections in amphibians; they may explain the vertical transmission of Goussia in tadpoles.

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

    Science.gov (United States)

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

  4. Adhesive-tape recovery combined with molecular and microscopic testing for the detection of Cryptosporidium oocysts on experimentally contaminated fresh produce

    Science.gov (United States)

    Cryptosporidium parvum suspended in water were applied to the surface of apples, cucumbers, peaches, and tomatoes at concentrations of 100, 50 or 10 oocysts within circles drawn with a permanent marker. Approximately 18 hr later, skin containing uncontaminated and contaminated circles from each prod...

  5. Hammondia heydorni: Oocyst shedding by dogs fed in vitro generated tissue cysts, and evaluation of cross-immunity between H. heydorni and Neospora caninum in mice.

    Science.gov (United States)

    Meneses, I D S; Schares, G; Rezende-Gondim, M M; Galvão, G S; Gondim, L F P

    2017-09-15

    Hammondia heydorni is a coccidian parasite believed to be nonpathogenic for naturally-infected animals, but it is biologically and genetically related to Neospora caninum, a worldwide cause of abortion in cattle. The major aim of the present work was to determine whether dogs shed H. heydorni oocysts after consuming in vitro generated tissue cysts of the parasite. In addition, we investigated cross-immunity between H. heydorni and N. caninum in mice. Two dogs were fed cultured cells containing tissue cysts of H. heydorni mixed with canned dog food, and a third dog (negative control) received only non-infected cells mixed with canned food. The two dogs that consumed in vitro produced tissue cysts shed high numbers of oocysts, which were induced to sporulate and tested positive for H. heydorni by a species-specific PCR. The third uninfected dog did not shed H. heydorni oocysts in the feces. Oocysts shed by the dogs induced the formation of encysted bradyzoites of H. heydorni on KH-R cells. Nineteen BALB/c mice were employed in the cross-immunity study. Nine mice were orally inoculated with 1×10(5) sporulated oocysts of H. heydorni and challenged with N. caninum tachyzoites 30days after infection with H. heydorni. Other ten mice, which did not receive H. heydorni oocysts, were infected with 2×10(5)N. caninum tachyzoites. Thirty days after challenging with N. caninum, all mice were euthanized and N. caninum DNA in their tissues was quantified by real time PCR. No statistically significant difference in N. caninum DNA concentrations were observed between the two groups. We concluded that in vitro generated cysts of H. heydorni are biologically active, because they induced oocyst shedding in dogs. As no cross-protection occurred in mice inoculated with H. heydorni and challenged with N. caninum, it is suspected that these parasites do not express significant numbers of homologous proteins during infection, or the immune response of BALB/c mice after H. heydorni

  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. Oocyst shedding by green-winged-saltator (Saltator similis) in the diagnostic of coccidiosis and Isospora similisi n. sp. (Apicomplexa: Eimeriidae).

    Science.gov (United States)

    Coelho, Cleide Domingues; Berto, Bruno Pereira; Neves, Daniel Medeiros; Oliveira, Vinícius Modesto de; Flausino, Walter; Lopes, Carlos Wilson Gomes

    2013-01-01

    Diurnal periodicity is a phenomenon that has been observed in coccidian of Isospora parasites of passerines, which have been eliminated great number of oocysts at dusk. The objective of this study was to evaluate the occurrence of periodicity of oocysts presence in the green-winged-saltator Saltator similis, and its use in the diagnosis of coccidiosis in wild birds in captivity. A total of 220 fecal samples were collected from birds, apprehended from illegal trading and kept in quarantine in CETAS∕IBAMA, in the morning and late afternoon, from May to November 2010. It was observed that 1.82% of the samples collected in the morning were positive, while 31.36% of samples were positive in the late afternoon. In addition, the number of oocysts shed was greater in the afternoon. Therefore, it was concluded that the sampling in the late afternoon provided greater reliability for the diagnosis of coccidiosis in green-winged-saltators. Moreover, in this study a new isosporoid coccidian parasite from the green-winged-saltator S. similis was observed and is herein described. Isospora similisi n. sp. oocysts are spheroidal to sub-spheroidal, 27.5 × 25.9 µm, with a smooth and bi-layered wall, ∼1.2 mm. Micropyle and oocyst residuum are absent, but splinter-like or comma-like granules are present. Sporocysts are ellipsoidal or slightly ovoidal, 17.4 × 12.2 mm. A stieda body and substieda body are present. The sporocyst residuum is composed of granules of different sizes. Sporozoites are vermiform with a single refractile body and a nucleus. This is the fourth description of an isosporoid coccidium infecting S. similis and the sixth description from Cardinalidae.

  8. 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.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2013-02-04

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

  10. Factors associated with the occurrence and level of Isospora suis oocyst excretion in nursing piglets of Greek farrow-to-finish herds

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    Skampardonis Vasilis

    2012-11-01

    Full Text Available Abstract Background Piglet isosporosis is one of the most common parasitic diseases in modern pig production. To prevent clinical disease, prophylactic treatment of piglets with toltrazuril (BAYCOX® 5%, Bayer HealthCare, Animal Health, Monheim, Germany is widely practiced in the past 20 years. There are only very few reports documenting the likely effect of managerial practices, such as hygiene measures, all-in-all-out management of farrowing facilities and piglet manipulations, and/or farm-specific environment - i.e. design and materials of the farrowing pen and room - in the risk of disease occurrence and transmission. Therefore, in this cross-sectional study, we identified litter- and herd-level factors associated with the odds and the level of Isospora suis oocyst excretion in nursing piglets of Greek farrow-to-finish pig herds. Faecal samples were collected from 314 liters of 55 randomly selected herds. Oocyst counts were determined by a modified McMaster technique and possible risk-factor data were collected through a questionnaire. In the analysis, we employed a two-part model that simultaneously assessed the odds and the level of oocyst excretion. Results Factors associated with lower odds of oocyst excretion were: use of toltrazuril treatment, all-in all-out management of the farrowing rooms, no cross-fostering or fostering during the first 24 hours after farrowing, plastic flooring in the farrowing pens, farrowing rooms with more than fourteen farrowing pens and employment of more than two caretakers in the farrowing section. Factors associated with lower oocyst excretion level were: use of toltrazuril treatment and caretakers averting from entering into farrowing pens. Conclusion Apart from prophylactic treatment with toltrazuril, the risk and the level of I. suis oocyst excretion from piglets in their second week of life, was associated with managerial and environmental factors. Changes in these factors, which may enhance prevention

  11. Occurrence of Cryptosporidial Oocysts and Giardia Cysts in Bottled Mineral Water Commercialized in the City of Campinas, State of São Paulo, Brazil

    Directory of Open Access Journals (Sweden)

    RMB Franco

    2002-03-01

    Full Text Available The consumption of bottled mineral water has significantly increased in Brazil so that it is in the interest of public health to determine the parasitological and microbiological status of some brands of Brazilian mineral water available in the town of Campinas, São Paulo, Brazil. For this purpose, detection of protozoa by direct immunofluorescence technique and microbiological parameters were determined for each specimen after membrane filtration. Giardia cysts were not present while cryptosporidial oocysts were detected in two samples. The counts of protozoa varied from 0.2 to 0.5 oocysts/l. The detected level of Pseudomonas aeruginosa and heterotrophic bacteria reflected the level of organic enrichment of the water.

  12. Effect of ingested human antibodies induced by RTS, S/AS01 malaria vaccination in children on Plasmodium falciparum oocyst formation and sporogony in mosquitoes

    DEFF Research Database (Denmark)

    Miura, Kazutoyo; Jongert, Erik; Deng, Bingbing

    2014-01-01

    BACKGROUND: The circumsporozoite protein (CS protein) on the malaria parasites in mosquitoes plays an important role in sporogony in mosquitoes. The RTS,S/AS01 malaria vaccine candidate, which has shown significant efficacy against clinical malaria in a large Phase 3 trial, targets the Plasmodium...... falciparum CS protein, but the ability of serum from vaccinated individuals to inhibit sporogony in mosquitoes has not been evaluated. METHODS: Previously a double-blind, randomized trial of RTS,S/AS01 vaccine, as compared with rabies vaccine, in five- to 17-month old children in Tanzania was conducted...... of antibodies to inhibit P. falciparum oocyst formation and/or sporogony in the mosquito host was evaluated by a standard membrane-feeding assay. The test antibodies were fed on day 0 (at the same time as the gametocyte feed), or on days 3 or 6 (serial-feed experiments). The oocyst and sporozoite counts were...

  13. [Occurrence of Cryptosporidium spp. oocysts (Apicomplexa, Cryptosporidiidae) in ostriches, Struthio camelus L., 1758 (Aves, Struthionidae) reared in North and Lowered Coastline regions of the state of Rio de Janeiro, Brazil].

    Science.gov (United States)

    Oliveira, Francisco Carlos R; Ederli, Nicole B; Ederli, Bianca B; Albuquerque, Marcia C; Dos Santos, Michelle Daniele

    2008-09-01

    The aim of this research was to determine the prevalence of Cryptosporidium spp. in ostriches reared in the state of Rio de Janeiro. The diagnosis of oocysts of Cryptosporidium spp. in the feces of 77 ostriches was done by modified Ziehl-Neelsen technique. The occurrence was observed in 44.4% of the examined animals. However, in a property 100% of the ostriches shed oocysts in their feces. Oocysts was subspherical in shape with 6.33 +/- 1.27 (4.31-9.63) by 5.90 +/- 1.18 (4.07-9.42) mm and shape index of 1.07 +/- 0.05 (1.00-1.29). Was verified a high occurrence of oocysts elimination. In spite was observed a uniform distribution of the oocysts measurements in the regression line (R2 = 0.9438) accentuated difference was evidenced in the morphometry of the oocysts, that suggests that more of a species of Cryptosporidium are parasitizing these birds.

  14. Differences in the detection of Cryptosporidium and Isospora (Cystoisospora) oocysts according to the fecal concentration or staining method used in a clinical laboratory.

    Science.gov (United States)

    Pacheco, Flávia T F; Silva, Renata K N R; Martins, Adson S; Oliveira, Ricardo R; Alcântara-Neves, Neuza M; Silva, Moacir P; Soares, Neci M; Teixeira, Márcia C A

    2013-12-01

    Despite the availability of many parasitological methods for detection of Cryptosporidium and Isospora (Cystoisospora) belli in fecal samples, there are uncertainties about the accuracy of these techniques in laboratory practice. In this study, 27 formalin-fixed positive stool samples for Cryptosporidium and 15 for I. belli were analyzed by 2 concentration methods, sedimentation by centrifugation (SC) and formalin-ethyl acetate (FE), and by 3 tintorial techniques, modified Ziehl-Neelsen (ZN), safranin (SF), and auramine (AR). No significant differences were observed on Cryptosporidium identification between concentration methods, while a significantly higher number of I. belli oocysts (P < 0.0001) was detected in fecal smears concentrated by the SC than by the FE method. Fecal samples processed by FE produced a median oocyst loss to the fatty ring of 34.8% for Cryptosporidium and 45.4% for I. belli. However, FE concentration provided 63% of Cryptosporidium and 100% of I. belli slides classified as superior for microscopic examination. Regarding the efficiency of staining methods, a more significant detection of Cryptosporidium oocysts was observed in fecal smears stained by ZN (P < 0.01) or AR (P < 0.05) than by the SF method. Regular to high-quality slides for microscopic examination were mostly observed in fecal smears stained with AR or ZN for Cryptosporidium and with SF or ZN for I. belli. This study suggests a great variability in oocyst power detection by routine parasitological methods, and that the most frequent intestinal coccidians in humans have specific requirements for concentration and staining.

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

    OpenAIRE

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

    2014-01-01

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

  16. 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

    2015-11-01

    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 gondii was 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.

  17. 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.

  18. Aflatoxin effect on the oocysts morphometry and contribution on the morphology of Eimeria bateri Bhatia, Pandey and Pande, 1965 from the japanese quail Coturnix japonica in Brazil Efeito da aflatoxina sobre a morfometria dos oocistos e contribuição para a morfologia de Eimeria bateri Bhatia, Pandey and Pande, 1965 da Codorna japonesa Coturnix japonica no Brasil

    OpenAIRE

    Bruno P. Berto; Sergian V. Cardozo; Walter L. Teixeira Filho; Ana Maria R. Ferreira; Lopes,Carlos Wilson G.

    2008-01-01

    The purpose of this study was to characterize Eimeria bateri oocysts and to evaluate the aflatoxin effect in the morphometry of sporulated oocysts in Japanese quails infected naturally. Of a total of 50 quails naturally infected by E. bateri were randomly divided into two groups with 25 birds each. In one of them, quails were orally administered with aflatoxin in dose of 0.04 mg/kg body weight previously. Both experimental groups shed E. bateri oocysts. These oocysts were subspherical to elli...

  19. Molecular identification of Neospora caninum from calf/foetal brain tissue and among oocysts recovered from faeces of naturally infected dogs in southern Ethiopia.

    Science.gov (United States)

    Asmare, K; Skjerve, E; Bekele, J; Sheferaw, D; Stachurska-Hagen, T; Robertson, L J

    2014-02-01

    This study sought to confirm and investigate further recently published information regarding the occurrence of Neospora caninum in cattle in Ethiopia and investigate infection in dogs, the canine definitive host, in this region. Faecal samples from 383 dogs in Hawassa, Ethiopia were examined by microscopy for Neospora-like oocysts, and positive samples then analysed by a molecular approach (DNA isolation, PCR and sequencing at the ITS1 gene). Brain tissue samples from four late term aborted foetuses, one congenitally defective calf (hind leg arthrogryposis) and placental tissue from cattle in the same area were also examined by the same molecular approach. All foetal, calf and placental tissue were associated with Neospora seropositive dams. A high prevalence of Neospora-like oocysts (11.5 μm±1.5 μm diameter) was observed in faecal samples from dogs (37 positive samples; 9.7% prevalence), and in 17 of these the identification was confirmed by PCR, giving a prevalence of confirmed infection of 4.4%. N. caninum DNA was also detected in all foetal and calf brain tissue samples. Sequencing revealed only minor differences among all PCR products, whether from oocysts or from brain tissue samples. These data provide molecular evidence of the presence of N. caninum infection in both dog and cattle in this region of Ethiopia. Moreover these findings highlight the role of dogs in maintaining and spreading the infection horizontally in the study area. The high frequency of N. caninum infection in household dogs as well as farm dogs is worthy of further investigation.

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

    Science.gov (United States)

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

    2017-06-01

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

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

    Science.gov (United States)

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

    2013-01-01

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

  2. 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.

  3. Therapeutic and Safety Evaluation of Combined Aqueous Extracts of Azadirachta indica and Khaya senegalensis in Chickens Experimentally Infected with Eimeria Oocysts

    Directory of Open Access Journals (Sweden)

    J. G. Gotep

    2016-01-01

    Full Text Available Coccidiosis is a disease of economic importance in poultry causing morbidity and mortality. Reports show that Azadirachta indica and Khaya senegalensis have been used individually in the treatment of avian coccidiosis. We thus investigated the efficacy and safety of the combined aqueous extracts of these plants for the treatment of experimentally induced coccidiosis in broiler chickens using oocyst count, oxidative stress biomarkers, serum biochemistry, histology, and haematological parameters. The phytochemical screening revealed the presence of tannins, saponins, cardiac glycosides, and steroids in both extracts. In addition, alkaloids and flavonoids were present in Azadirachta indica. There was significant (p<0.05 dose dependent decrease in oocyst count across the treatment groups with 400 mg/kg of the combined extract being the most efficacious dose. Immunomodulatory and erythropoietic activity was observed. There were decreased intestinal lesions and enhanced antioxidant activity across the treatment groups compared to the negative control. Administration of the combined extract did not cause damage to the liver as ALT, AST, and ALP levels were significantly reduced in the uninfected chickens treated with the extracts compared to control suggesting safety at the doses used. The combined aqueous extracts of K. senegalensis stem bark and Azadirachta indica leaves were ameliorative in chickens infected with coccidiosis.

  4. Therapeutic and Safety Evaluation of Combined Aqueous Extracts of Azadirachta indica and Khaya senegalensis in Chickens Experimentally Infected with Eimeria Oocysts

    Science.gov (United States)

    Gotep, J. G.; Tanko, J. T.; Forcados, G. E.; Muraina, I. A.; Ozele, N.; Dogonyaro, B. B.; Oladipo, O. O.; Makoshi, M. S.; Akanbi, O. B.; Kinjir, H.; Samuel, A. L.; Onyiche, T. E.; Ochigbo, G. O.; Aladelokun, O. B.; Ozoani, H. A.; Viyoff, V. Z.; Dapuliga, C. C.; Atiku, A. A.; Okewole, P. A.; Shamaki, D.; Ahmed, M. S.; Nduaka, C. I.

    2016-01-01

    Coccidiosis is a disease of economic importance in poultry causing morbidity and mortality. Reports show that Azadirachta indica and Khaya senegalensis have been used individually in the treatment of avian coccidiosis. We thus investigated the efficacy and safety of the combined aqueous extracts of these plants for the treatment of experimentally induced coccidiosis in broiler chickens using oocyst count, oxidative stress biomarkers, serum biochemistry, histology, and haematological parameters. The phytochemical screening revealed the presence of tannins, saponins, cardiac glycosides, and steroids in both extracts. In addition, alkaloids and flavonoids were present in Azadirachta indica. There was significant (p < 0.05) dose dependent decrease in oocyst count across the treatment groups with 400 mg/kg of the combined extract being the most efficacious dose. Immunomodulatory and erythropoietic activity was observed. There were decreased intestinal lesions and enhanced antioxidant activity across the treatment groups compared to the negative control. Administration of the combined extract did not cause damage to the liver as ALT, AST, and ALP levels were significantly reduced in the uninfected chickens treated with the extracts compared to control suggesting safety at the doses used. The combined aqueous extracts of K. senegalensis stem bark and Azadirachta indica leaves were ameliorative in chickens infected with coccidiosis. PMID:26989496

  5. Quantification of Eimeria acervulina in faeces of broilers: Comparison of McMaster oocyst counts from 24 h faecal collections and single droppings to real-time PCR from cloacal swabs

    NARCIS (Netherlands)

    Velkers, F.C.; Blake, D.P.; Graat, E.A.M.; Vernooij, J.C.M.; Bouma, A.; de Jong, M.C.M; Stegeman, J.A.

    2010-01-01

    Coccidiosis is an economically important disease in chickens, caused by infection with Eimeria species parasites. Diagnosis of coccidiosis is frequently based on oocyst enumeration in pooled faecal samples or litter. In studies on infection dynamics and for monitoring in the field, samples from indi

  6. Eimeria uz ura Single Oocyst Isolation and Amplification in Young Quails%日本鹌鹑艾美耳球虫单卵囊分离及雏鹑扩增研究

    Institute of Scientific and Technical Information of China (English)

    张敏; 韩琼琼; 李清州; 王芳; 梁楠; 张龙现; 菅复春; 宁长申

    2014-01-01

    To obtain a large number of pure-bred E .uz ura ,quails’ coccidia oocysts were collected using the saturated brine flotation method and cultured at 26 ℃ .According to morphological identification , single sporulated oocyst of E .uzurawas separated and amplified in young quails .The results showed that sporulation of oocysts occurred for 14 h at 26 ℃ .seven-day-old artificially-reared coccidian-free quails were inoculated orally with 1 and 200 sporulated oocysts of E .uz ura ,the prepatent period was 6 d and 5 d ,and discharge of oocysts continued for 8-10 d and 11 d ,respectively .%为获取大量纯种日本鹌鹑艾美耳球虫(E .uz ura),采用饱和食盐水漂浮法收集某场鹌鹑的球虫卵囊,26℃恒温培养至孢子化后,对据形态学鉴定为 E .uz ura的分离株进行单卵囊分离和雏鹑扩增试验。结果显示,26℃条件下卵囊孢子化时间为14 h ;单卵囊感染7日龄雏鹑,其潜隐期6 d ,显露期8~10 d;以200个卵囊感染7日龄雏鹑,其潜隐期为5 d ,显露期为11 d。

  7. Toxoplasmosis: The spread of tachyzoites, cysts and oocysts%弓形虫病:速殖子、包囊和卵囊的传播

    Institute of Scientific and Technical Information of China (English)

    汪涛; 汤自豪

    2012-01-01

    Toxo plasma gondii is widely distributed opportunistic pathogenic within the world, which can infect all warm-blooded animals including human and cause zoonotic toxoplasmosis , seriously throating human health and livestock production . This article focuses on the biological characteristics and transmission feature of Toxop lasma gondii tachyzoite , cysts and oocysts.%目的 弓形虫是一种世界范围内广泛分布的机会致病原虫,能感染包括人在内的所有温血动物,能引起人兽共患弓形虫病,严重威胁着人类健康和畜牧业生产.本文重点阐述弓形虫速殖子、包囊和卵囊的生物学特性和传播特点.

  8. Faecal helminth egg and oocyst counts of a small population of African lions (Panthera leo in the southwestern Kalahari, Namibia : research communication

    Directory of Open Access Journals (Sweden)

    Y. Smith

    2006-09-01

    Full Text Available An endoparasite survey of a small pride of African lions (Panthera leo was conducted at Intu Afrika Kalahari Game Reserve, southwestern Namibia, during winter and summer of 2003 and 2004, respectively. Overall, 23 fresh lion scats were collected opportunistically during fieldwork trials. A flotation technique was employed for the diagnosis of parasites. Three nematodes, Ancylostoma braziliense, Gnathostoma spinigerum and Uncinaria stenocephala and two coccidians, Toxoplasma gondii and Isospora felis were recorded. By using the McMaster method for quantification, a maximum number of 14 866 oocysts per gram of faeces was obtained for I. felis during winter 2003. Endoparasite taxa carried by the different individuals in the pride were found to be related to their levels of association. Rates of infection were relatively low as a result of the habitat, semi-captive conditions and earlier sporadic deworming.

  9. Protocol for DNA extraction of Cryptosporidium spp. oocysts in fecal samples Protocolo para extração de DNA de oocistos de Cryptosporidium spp. em amostras fecais

    Directory of Open Access Journals (Sweden)

    Elenice M.N. Gonçalves

    2008-06-01

    Full Text Available Molecular characterization of Cryptosporidium spp.oocysts in clinical samples is useful for public health since it allows the study of sources of contamination as well as the transmission in different geographical regions. Although widely used in developed countries, in Brazil it is restricted to academic studies, mostly using commercial kits for the extraction of genomic DNA, or in collaboration with external reference centers, rendering the method expensive and limited. The study proposes the application of the modifications recently introduced in the method improving feasibility with lower cost. This method was efficient for clinical samples preserved at -20 °C for up to six years and the low number of oocysts may be overcomed by repetitions of extraction.A caracterização molecular de oocistos de Cryptosporidium spp. em amostras clínicas é útil à saúde pública, pois permite estudo das fontes de contaminação e a transmissão em determinadas regiões geográficas. Apesar de largamente utilizada em países desenvolvidos, no Brasil está restrita aos estudos acadêmicos, na maioria utilizando kits comerciais para extração do DNA genômico, ou em colaborações com centros de referência externos, o que torna o método caro e limitado. Este estudo propõe a introdução de modificações nos métodos existentes para melhorar a viabilidade e baixar custos. O método proposto foi eficiente em amostras clínicas preservadas a -20 °C por até seis anos e o baixo número de oocistos pode ser contornado por replicadas extrações de DNA.

  10. Effects of vaccination against coccidiosis, with and without a specific herbal essential oil blend, on performance, oocyst excretion and serum IBD titers of broilers reared on litter

    Directory of Open Access Journals (Sweden)

    Mustafa Çınar

    2012-01-01

    Full Text Available The aim of the present experiment was to investigate the effects of oral administration of a live attenuated vaccine (VAC and an essential oil blend (EOB, either alone or in combination, as a novel anticoccidial strategy for broiler chickens with a mixed Eimeria spp. infection. A total of 624 broiler chicks were randomly assigned to one of six treatments. Two of the groups, only one of which was challenged with coccidiosis, were given a basal diet and served as controls. The other two groups, also infected, were given a basal diet supplemented with monensin sodium (MON, 100 mg/kg or the EOB (75 mg/kg. Of the remaining two groups, which were infected with coccidiosis, one was vaccinated against coccidiosis (VAC and the other was both vaccinated and fed a diet with an EOB (VAC+EOB. Birds treated with VAC and VAC+EOB had comparable live performance to MON-fed birds challenged with coccidiosis. Conversely, EOB diet supplementation had negative effects on growth, feed intake and feed conversion ratio throughout the growth period. None of the coccidial control strategies affected the overall performance of uninfected birds. There was no significant difference in mortality among treatments. All of the anticoccidial procedures kept serum infectious bursal disease titers at high levels after coccidial infection and reduced fecal oocyst excretion, with the exception of the MON-based procedure. The results indicate that vaccination against coccidiosis, with or without EOB, demonstrated the same efficacy in promoting recovery from coccidial infection and in reducing oocyst shedding as MON.

  11. The Extermination Effect of Pomegranate Peel Extracts on Rabbit Coccidian Oocysts Cultured in Vitro%石榴皮提取物对离体培养兔球虫卵囊的杀灭作用

    Institute of Scientific and Technical Information of China (English)

    王玺德; 盛书娟; 祝建平; 崔希云; 谢德颖

    2012-01-01

    The exterminatory actions of pomegranate peel extract to rabbit coccidia oocysts cultured in vitro were studied in this paper. We collected the excrement (24 h, 35 g each rabbit) from eighteen rabbits infected with coccidian, weighing 2. 0 to 2. 5 kg, separated and extracted the coccidia oocysts with saturation salt water floating method. The coccidia oocysts were cultivated in distilled water, aqueous extract of pomegranate peels, pomegranate flavonoids, pomegranate polyphenols and Robe-nidine HC1 for 24, 48 and 78 h, respectively. The number of rabbit coccidia oocysts was observed with the optical microscope. The effects of different concentrations of drugs and different cultivated time on the amount of rabbit coccidia oocysts were estimated. The results showed that comparing with distilled water group, the amount of rabbit coccidia oocysts separately cultivated in 72 h and 5,10 mg of the four drugs groups reduced, and the reducing number ranged in descending order was aqueous extract of pomegranate peels group, pomegranate flavonoids group, pomegranate polyphenols group and Robebidine HC1 group. The onset time of the extracts of pomegranate peel were 48 and 72 h. The amount of rabbit coccidia oocysts cultivated in 24, 48, 72 h and 1 mg of the four drugs groups were not reduced significantly (P>0. 05). There were many notes which indicated that the extracts of pomegranate peel were the stronger exterminatory action to the rabbit coccidia oocysts cultured in vitro. The exterminatory action of aqueous extract to the rabbit coccidia oocysts was the strongest, followed by pomegranate polyphenols and pomegranate flavonoids in the 3 extractions from pomegranate peel. The exterminatory action had a certain positive correlation with the dose.%本试验旨在研究石榴皮提取物对离体培养兔球虫卵囊的杀灭作用.采集已感染球虫的18只家兔(体重2.0~2.5kg)的新鲜粪便(24 h内,35 g/只),用饱和盐水漂浮法分离、提取球虫卵囊.球虫

  12. Single-oocyst Separation and Identification of Eimeria tenella in Chickens in Patrs of Anhui Province%安徽部分地区鸡柔嫩艾美耳球虫单卵囊的分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    王中宝; 欧俊秀; 曾明华; 阮祥春; 张伟玮; 杨娜; 吴丽君

    2013-01-01

    In order to obtain pure Eimeria strain, three Eimeria species were collected from the poultry farms infected with coccidia in Chaohu, Huoqiu and Dingyuan of Anhui Province, and then cultured to sporu-lated oocysts. The 1-7 days old chicks were vaccinated the single-oocyst by single-oocyst separation technology. The oocysts were collected after 7-12 days, and then the coccidia oocysts were identified with biological characteristics and PCR method. The results showed that the ratio of infection were 50%, 57% and 29%, respectively in three areas. The three Eimeria species were all identified as Eimeria tenella by biological characteristics and PCR. This indicated that the single oocyst isolation technique was simply and successful. Using specific primers of PCR to identify strains could lay foundation for the study of pure local strains.%为获得纯种柔嫩艾美耳球虫虫株,从安徽省巢湖市、霍邱县、定远县3个地区患球虫病的鸡场采集3株混合球虫,培养至孢子化,分离单卵囊接种1~7日龄的雏鸡,感染后7~12d,收集卵囊,然后通过生物学特性和PCR方法进行虫种鉴定.结果显示,3个地区的单卵囊感染成功率分别为50%、57%和29%,各虫株经生物学特性和PCR鉴定均为柔嫩艾美耳球虫.研究表明:1.0%琼脂单卵囊分离法简便且成功率较高,利用特异性引物PCR鉴定虫株,为纯种地方株的研究奠定了基础.

  13. Structure and function of a G-actin sequestering protein with a vital role in malaria oocyst development inside the mosquito vector.

    Science.gov (United States)

    Hliscs, Marion; Sattler, Julia M; Tempel, Wolfram; Artz, Jennifer D; Dong, Aiping; Hui, Raymond; Matuschewski, Kai; Schüler, Herwig

    2010-04-09

    Cyclase-associated proteins (CAPs) are evolutionary conserved G-actin-binding proteins that regulate microfilament turnover. CAPs have a modular structure consisting of an N-terminal adenylate cyclase binding domain, a central proline-rich segment, and a C-terminal actin binding domain. Protozoan parasites of the phylum Apicomplexa, such as Cryptosporidium and the malaria parasite Plasmodium, express small CAP orthologs with homology to the C-terminal actin binding domain (C-CAP). Here, we demonstrate by reverse genetics that C-CAP is dispensable for the pathogenic Plasmodium blood stages. However, c-cap(-) parasites display a complete defect in oocyst development in the insect vector. By trans-species complementation we show that the Cryptosporidium parvum ortholog complements the Plasmodium gene functions. Purified recombinant C. parvum C-CAP protein binds actin monomers and prevents actin polymerization. The crystal structure of C. parvum C-CAP shows two monomers with a right-handed beta-helical fold intercalated at their C termini to form the putative physiological dimer. Our results reveal a specific vital role for an apicomplexan G-actin-binding protein during sporogony, the parasite replication phase that precedes formation of malaria transmission stages. This study also exemplifies how Plasmodium reverse genetics combined with biochemical and structural analyses of orthologous proteins can offer a fast track toward systematic gene characterization in apicomplexan parasites.

  14. Effects of lasalocid and monensin in combination with roxarsone on lesion reduction and oocyst suppression in chicks infected with Eimeria tenella field isolates.

    Science.gov (United States)

    Mitrovic, M; Schildknecht, E; Trainor, C

    1977-05-01

    The anticoccidial activity of lasalocid, monensin, and roxarsone, alone and in combination, was evaluated against eleven Eimeria tenella recent field isolates. Lasalocid was used at 0.0075. 0.01, and 0.0125% activity drug in feed; monensin at 0.0099 and 0.0121%; and roxarsone at 0.005%. Further studies with lasalocid 0.0075%, monensin 0.0099% and roxarsone 0.005 and 0.0025% combinations were carried out against three E. tenella field isolates selected from the aforementioned strains. Lasalocid and monensin each exhibited a high degree of anticoccidial activity at all concentrations tested. Lasalocid and monensin fed in combination with roxarsone showed, in addition to high anticoccidial activity a further reduction in gross lesions and oocysts production, more pronounced at 0.005% level of roxarsone than at 0.0025%, compared to either medication alone or the roxarsone combinations. These positive effects were noted with all strains tested. The practical aspects of these findings are discussed.

  15. Campylobacter spp., Enterococcus spp., Escherichia coli, Salmonella spp., Yersinia spp., and Cryptosporidium oocysts in semi-domesticated reindeer (Rangifer tarandus tarandus in Northern Finland and Norway

    Directory of Open Access Journals (Sweden)

    Aschfalk A

    2006-06-01

    Full Text Available Abstract The specific aim of this study was to assess the faecal shedding of zoonotic enteropathogens by semi-domesticated reindeer (Rangifer tarandus tarandus to deduce the potential risk to human health through modern reindeer herding. In total, 2,243 faecal samples of reindeer from northern regions of Finland and Norway were examined for potentially enteropathogenic bacteria (Campylobacter species, Enterococcus species, Escherichia coli, Salmonella species and Yersinia species and parasites (Cryptosporidium species in accordance with standard procedures. Escherichia coli were isolated in 94.7%, Enterococcus species in 92.9%, Yersinia species in 4.8% of the samples and Campylobacter species in one sample only (0.04%. Analysis for virulence factors in E. coli and Yersinia species revealed no pathogenic strains. Neither Salmonella species nor Cryptosporidium oocysts were detected. The public health risk due to reindeer husbandry concerning zoonotic diseases included in this study has to be considered as very low at present but a putative epidemiological threat may arise when herding conditions are changed with respect to intensification and crowding.

  16. Comparison of host cell invasion and proliferation among Neospora caninum isolates obtained from oocysts and from clinical cases of naturally infected dogs.

    Science.gov (United States)

    Dellarupe, A; Regidor-Cerrillo, J; Jiménez-Ruiz, E; Schares, G; Unzaga, J M; Venturini, M C; Ortega-Mora, L M

    2014-10-01

    In a previous study we have shown that the in vitro invasion rate (IR) and tachyzoite yield (TY) are associated with the virulence phenotypes of Neospora caninum isolates of bovine origin. In addition, we recently observed marked differences in virulence when canine isolates were compared in a pregnant BALB/c mouse model. In this study, we investigated whether invasion and proliferation capacities could be used as virulence-related N. caninum phenotypic traits. Of the isolates compared in mice, four canine isolates obtained from oocysts (Nc-Ger2, Nc-Ger3, Nc-Ger-6, Nc-6 Arg) had shown a low-moderate virulence, and two further isolates obtained from dogs with neurological signs (Nc-Bahia, Nc-Liv) were highly virulent. The IR for each isolate was determined by a plaque assay and the counting of immunofluorescence-labeled parasitophorous vacuoles at 3 days post-inoculation (p.i.). The TY was determined by the quantification of tachyzoites at 56 h p.i. by real-time PCR. Most of the canine isolates showed similar IR values under controlled invasion conditions for 4h and 72 h p.i., indicating a limited time period for invasion similar to that observed for bovine isolates. The Nc-Ger3, Nc-Bahia, and Nc-Liv isolates showed a significantly higher IR and TY than the Nc-Ger2 and Nc-Ger6 isolates (P0.885, P0.8117, P<0.058) in mice. These results demonstrate the importance both the invasive and proliferative capacities have on the virulence of canine N. caninum isolates.

  17. Spatial distribution of soil contaminated with Toxoplasma gondii oocysts in relation to the distribution and use of domestic cat defecation sites on dairy farms.

    Science.gov (United States)

    Simon, J A; Kurdzielewicz, S; Jeanniot, E; Dupuis, E; Marnef, F; Aubert, D; Villena, I; Poulle, M-L

    2017-03-16

    Little information is available on the relationship between the spatial distribution of zoonotic parasites in soil and the pattern of hosts' faeces deposition at a local scale. In this study, the spatial distribution of soil contaminated by the parasite Toxoplasma gondii was investigated in relation to the distribution and use of the defecation sites of its definitive host, the domestic cat (Felis catus). The study was conducted on six dairy farms with a high number of cats (seven to 30 cats). During regular visits to the farms over a 10month period, the cat population and cat defecation sites (latrines and sites of scattered faeces) on each farm were systematically surveyed. During the last visit, 561 soil samples were collected from defecation sites and random points, and these samples were searched for T. gondii DNA using real-time quantitative PCR. Depending on the farm, T. gondii DNA was detected in 37.7-66.3% of the soil samples. The proportion of contaminated samples at a farm was positively correlated with the rate of new cat latrines replacing former cat latrines, suggesting that inconstancy in use of a latrine by cats affects the distribution of T. gondii in soil. On the farms, known cat defecation sites were significantly more often contaminated than random points, but 25-62.5% of the latter were also found to be contaminated. Lastly, the proportion of positive T. gondii samples in latrines was related to the proximity of the cats' main feeding and resting sites on the farms. This study demonstrates that T. gondii can be widely distributed in farm soil despite the heterogeneous distribution of cat faeces. This supports the hypothesis that farms are hotspot areas for the risk of T. gondii oocyst-induced infection in rural environments.

  18. The immunogenicity of Eimeria tenella sporozoite proteins and living oocyst vaccines in broilersImunogenicidade de vacina com proteínas de esporozoítos e oocistos vivos de Eimeria tenella em frangos

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    Odilon Vidotto

    2012-02-01

    Full Text Available The purpose of the present study was to evaluate the immunity of broilers inoculated with sporulated oocysts and sporozoite proteins of Eimeria tenella against challenge with homologous infectives oocysts. Broiler chickens of Hubbard strain, females, coccidian-free, were kept in wire cages and inoculated on the third day (day 0. Three treatments were used: T1, the negative control; T2, received 500 sporulated oocysts by gavage; T3, positive control; T4, received 50 ?g of sporozoite protein with Quil A; and T5, received Quil A without sporozoite protein and PBS, the last two protocols were administered via nasal route on days 0, 7, and 21. On the 31st day, all groups, with the exception of T1, were challenged with homologous virulent strain of E. tenella at the dose of 8.0×104 oocysts. Immunogenicity was assessed by carotenoids, ELISA assay, histopathological analysis, oocysts count and lesion score. Antibody kinetics were measured weekly and showed a gradual increase in levels of immunoglobulin for treatment T2 and T4, reaching a peak on day 21. Based on the parameters evaluated, a total protecting immunological effect for the infectant oocysts vaccine (T2 and partial protection for the sporozoite protein vaccine (T4 against homologous virulent strain challenge were observed. O objetivo do presente estudo foi avaliar a imunidade de frangos de corte inoculados com oocistos esporulados e proteínas de esporozoítos de Eimeria tenella utilizando um desafio com oocistos esporulados homólogos. Frangos de corte da linhagem Hubbard, fêmeas, livres de coccidiose, foram mantidos em gaiolas de arame e inoculados no terceiro dia (dia 0. O delineamento experimental consistiu de cinco tratamentos com três repetições: T1- controle negativo; T2- recebeu 500 oocistos esporulados por gavagem; T3- controle positivo; T4- receberam 50 ?g de proteína de esporozoítos adicionados a Quil-A, e finalmente o T5- que recebeu apenas Quil A e PBS, os últimos dois

  19. 猪源隐孢子虫卵囊的分离及基因型鉴定%Isolation and genotype identification of Cryptosporidium oocysts in pigs

    Institute of Scientific and Technical Information of China (English)

    陈甫; 黄克和

    2011-01-01

    To investigate the genotypes of Cryptosporidium parasiting in pigs in Nanjing,Shanghai and Hefei, the total RNA of the oocysts from 6 positive feces was extracted using the TRIZOL reagent,and the partial genes of the small subunit ribosomal RNA (SSU rRNA) were amplified by the reverse transcription polymerase chain reaction (RT-PCR). SSU rRNA gene fragments of the isolates were deposited in the GenBank database under accession NO. DQ855266,DQ855267 and compared with sequences of various Cryptosporidium spp. and genotypes on the NCBI server using the BLAST and DNAStar software by the analysis of the homologous rate and phylogenetic tree. The results showed that the SSU rRNA gene fragments,compared with sequences of various Cryptosporidium spp. and genotypes,belonged to the species C. parvum with 94 %-100 % homologous rate, and were most similar to the C. parvum “mouse” genotype with 99. 8%-100 % homologous rate. The phylogenetic tree and sequence distances gen erated showed that those isolates belong to the C. parvum groups,and lie in the same divarication with C. parvum “mouse” genotype. It is concluded that those isolates are clearly distinct from other sequences as to merit species status, which is C. parvum “mouse” genotype from pigs. There might be a cross-transmission between pigs and mice.%应用RT-PCR方法对南京、上海和合肥猪源隐孢子虫卵囊SSU rRNA部分序列进行扩增,产物测序后提交GenBank,收录号为DQ855266、DQ855267;用BLAST和DNAStar软件与GenBank参考序列进行比较,分析其同源性,绘制系统发育进化树,结合卵囊形态学观察和对小鼠、大鼠、兔、山羊和鸡的传染性试验确定隐孢子虫种类或基因型.结果表明,3地区猪源隐孢子虫分离株与微小隐孢子虫(C.parvum)同源性达94%~100%,与C.parvum"mouse"型有99.8%~100%的同源性,并处于进化树的同一分支.因此,3地区猪源隐孢子虫是C.parvum"mouse"型,提示猪和鼠之间存在交叉传播的可能.

  20. Diagnostic value of a Rec-ELISA using Toxoplasma gondii recombinant SporoSAG, BAG1, and GRA1 proteins in murine models infected orally with tissue cysts and oocysts.

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    Mert Döşkaya

    Full Text Available Toxoplasma gondii causes congenital toxoplasmosis in newborns resulting with fetal anomalies. Determining the initiation time of infection is very important for pregnant women and current serological assays have drawbacks in distinguishing the recently acute toxoplasmosis. Diagnosis of recently acute infection may be improved by using stage specific antigens in serological assays. In the present study, the diagnostic value of sporozoite specific SporoSAG, bradyzoite specific BAG1 proteins and GRA1 protein expressed by all forms of the parasite have been evaluated ELISA using sera systematically collected from mice administered orally with tissue cyst and oocysts. The anti-SporoSAG IgM antibodies in sera obtained from mice infected with oocysts peaked significantly at days 1, 10, and 15 (P<0.01. The anti-BAG1 IgM antibodies in sera obtained from mice infected with tissue cysts peaked significantly at days 15, 40, and 120 (P<0.05. The anti-GRA1 IgM antibodies in sera obtained from mice infected with oocysts peaked significantly at days 2, 10, and 40 (P<0.01. The anti-GRA1 IgM antibodies in sera obtained from mice infected with tissue cysts peaked significantly only at day 40 (P<0.05. The anti-SporoSAG, anti-BAG1, and anti-GRA1 IgG titers of mice showed significant increases at day 40 (P<0.05 and decrement started for only anti-GRA1 IgG at day 120. The presence of anti-SporoSAG IgM and IgG antibodies can be interpreted as recently acute infection between days 10-40 because IgM decreases at day 40. Similarly, presence of anti-BAG1 IgM and absence of IgG can be evaluated as a recently acute infection that occurred 40 days before because IgG peaks at day 40. A peak in anti-GRA1 antibody level at first testing and reduction in consecutive sample can be considered as an infection approximately around day 40 or prior. Overall, recombinant SporoSAG, BAG1 and GRA1 proteins can be accepted as valuable diagnostic markers of recently acute toxoplasmosis.

  1. APPLICATION OF COMPUTER TECHNOLOGY FOR EXAMINATION OF COCCIDIAN OOCYSTS IN SURVEY OF GOAT COCCIDIUM SPECIES%山羊球虫种类调查与电脑技术应用于卵囊测量方法的探索

    Institute of Scientific and Technical Information of China (English)

    阮正祥; 陈兴忠; 何雪锋; 刘君; 王芸华

    2011-01-01

    为探索在电脑显示屏上直接观察、记录和测量球虫卵囊的方法,作者采用漂浮法、麦克马斯特氏法、卵囊培养等方法,并应用电脑和配置有彩色数码摄像机的双目光学显微镜设备,对毕节市的122份山羊粪样进行调查,在电脑显示屏上观察、记录和测量球虫卵囊。结果显示,球虫卵囊的感染率为96.72%,OPG值为600~23 400枚,平均70 57枚,鉴别出艾美耳属球虫8种,并对其卵囊的形态学特征进行了详细地描述。本调查调查结果表明在电显示屏上直接观察、记录和测量球虫卵囊的方法比在显微镜下易于观察鉴别,测量也准确、方便和省力。%A total of 122 goat fecal specimens were collected in Bijie city of Guizhou Province and examined for the presence of coccidian oocysts by using floating method, McMaster's method and oocyst cultures. A color digital camera with binocular optical microscope was linked to a computer so that observation, recording and measurement of coccidian oocysts could be completed on the computer screen. The positive rate of coccidian oocysts was 96.72% and OPG ranged from 600 to 23400 (average 7057). Eimeria tenella was identified in 8 fecal specimens and the morphology of their oocysts was described. These results demonstrated the feasibility of direct observation, recording and measurement of coccidia oocysts on computer screen linked with color digital camera with binocular optical microscope.

  2. Produção irregular e inconstante de oocistos pela ministração de cistos de Toxoplasma gondii Nicole & Manceaux, 1909, em gatos Irregular and inconstant production of Toxoplasma gondii oocysts in cats by ministration of cysts

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    F. Nery-Guimarães

    1973-01-01

    Full Text Available Como a produção de oocistos pelo T. gondii no gato parece vinculada à forma cística, uma vez que ela não ocorre pela ministração de formas vegetativas (17, cérebros de camundongos e ratos, com infecção crônica de T. gondii, forma ministrados "per os" a um total de 32 gatos. Com uma amostra (TC1 nenhum dos 8 gatos experimentados eliminou oocistos atribuiíveis a T. gondii; e com as outras 3 amostras a produção de oocistos foi de 25% ("pombo" e 37,5% ("sonia" e "AS-28". Esses resultados forma atribuídos, em parte, à pobreza de cistos nos cérebros ministrados aos gatos; e, em parte, à existência em condições naturais de amostras de "T. gondii" oocistogênicas, paucloocistogênicas e noocistogênicas. Ao contrário dos gatos infectados com formas vegetativas (17, nenhum gato morreu de toxoplasmose; microscopicamente as lesões com toxoplasmas foram de pouca intensidade e o número de reações de Sabin-Feldman (RSF negativas foi, relativamente, elevado. Foram vistas raras formas evolutivas do T. gondii no epitélio intestinal de um gato, não parasitado por Isospora as quais são menores que as de I. felis e I. rivolta, do mesmo modo que os seus oocistos. Assim como parece não existir imunidade entre as Isospora (I. felis e I. rivolta, também parece não existir imunidade entre estas e o T. gondii. Em 185 gatos examinados, foi encontrado um deles com oocistos de T. gondii em condições naturais (0,54%. É discutida a importância do gato na epidemilogia da toxoplasmose, destacando-se que a descoberta do ciclo sexuado, decisiva para a classificação do parasito, parece não ter tido repercussão semelhante na epidemiologia.As the production of oocysts by T. gondii in the cat seems to be correlated with the cystic form, once it does not occur when vegetative forms are given to cats, (17, brains of mice and rats with T. gondii chronic infection were orally administered to 32 cats. With TC1 strain none of the eight cats

  3. Salivary IgA against sporozoite-specific embryogenesis-related protein (TgERP) in the study of horizontally transmitted toxoplasmosis via T. gondii oocysts in endemic settings.

    Science.gov (United States)

    Mangiavacchi, B M; Vieira, F P; Bahia-Oliveira, L M G; Hill, D

    2016-09-01

    The aim of this study was to contribute to the better understanding of the relative epidemiological importance of different modes of infection with respect to horizontal transmission of Toxoplasma gondii in endemic settings. We investigated the prevalence of salivary IgA against a sporozoite-specific embryogenesis-related protein (TgERP) in a highly endemic area for toxoplasmosis in Brazil in order to pinpoint parasite transmission via oocysts. Prevalence calculated by salivary IgA specific to TgERP was compared to the prevalence calculated by serum IgG against both TgERP and tachyzoites (in conventional serological tests). Prevalence calculated by different serological and salivary parameters varied in the studied age groups. However, for the 15-21 years age group, values for T. gondii prevalence estimated by conventional serological tests and by anti-TgERP salivary IgA were similar; i.e. 68·7% and 66·6% or 66·7%, respectively, using two different cut-off parameters for salivary IgA anti-TgERP. Furthermore, salivary IgA anti-TgERP for this age group presented the highest specificity (93·33%), sensitivity (93·94%), and likelihood (14·09) compared to all the other age groups. These data demonstrate the importance of age for salivary IgA investigation against TgERP to estimate the mode of T. gondii transmission in endemic settings.

  4. Oocyst shedding by green-winged-saltator (Saltator similis in the diagnostic of coccidiosis and Isospora similisi n. sp. (Apicomplexa: Eimeriidae Eliminação de oocistos por trinca-ferro-verdadeiro (Saltator similis no diagnóstico da coccidiose e Isospora similisi n. sp. (Apicomplexa: Eimeriidae

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    Cleide Domingues Coelho

    2013-03-01

    Full Text Available Diurnal periodicity is a phenomenon that has been observed in coccidian of Isospora parasites of passerines, which have been eliminated great number of oocysts at dusk. The objective of this study was to evaluate the occurrence of periodicity of oocysts presence in the green-winged-saltator Saltator similis, and its use in the diagnosis of coccidiosis in wild birds in captivity. A total of 220 fecal samples were collected from birds, apprehended from illegal trading and kept in quarantine in CETAS∕IBAMA, in the morning and late afternoon, from May to November 2010. It was observed that 1.82% of the samples collected in the morning were positive, while 31.36% of samples were positive in the late afternoon. In addition, the number of oocysts shed was greater in the afternoon. Therefore, it was concluded that the sampling in the late afternoon provided greater reliability for the diagnosis of coccidiosis in green-winged-saltators. Moreover, in this study a new isosporoid coccidian parasite from the green-winged-saltator S. similis was observed and is herein described. Isospora similisi n. sp. oocysts are spheroidal to sub-spheroidal, 27.5 × 25.9 µm, with a smooth and bi-layered wall, ∼1.2 mm. Micropyle and oocyst residuum are absent, but splinter-like or comma-like granules are present. Sporocysts are ellipsoidal or slightly ovoidal, 17.4 × 12.2 mm. A stieda body and substieda body are present. The sporocyst residuum is composed of granules of different sizes. Sporozoites are vermiform with a single refractile body and a nucleus. This is the fourth description of an isosporoid coccidium infecting S. similis and the sixth description from Cardinalidae.A periodicidade diurna é um fenômeno que tem sido observado em coccídios do gênero Isospora parasitas de pássaros, os quais eliminam uma maior quantidade de oocistos ao entardecer. O objetivo deste estudo foi determinar a periodicidade de eliminação de oocistos pelas fezes no trinca

  5. Systemic inflammatory response indicators in rabbits (Oryctolagus cuniculus experimentally infected with sporulated oocysts of Eimeria stiedai (Apicomplexa: Eimeriidae Indicadores da resposta inflamatória sistêmica em coelhos (Oryctolagus cuniculus infectados experimentalmente com oocistos esporulados de Eimeria stiedai (Apicomplexa: Eimeriidae

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    Fagner Luiz da Costa Freitas

    2011-06-01

    Full Text Available Hemograms and acute-phase proteins in adult male New Zealand White rabbits that had been experimentally infected orally with sporulated oocysts of Eimeria stiedai were evaluated over a 28-day period. Fifty animals were used, divided into two groups: group A infected with 1 × 10(4 sporulated oocysts of E. stiedai and group B inoculated with distilled water. On the seventh day after infection, the infected animals presented anemia and leukocytosis with neutrophilia and monocytosis. Protein fractionation by means of electrophoresis identified 19 acute-phase proteins with molecular weights ranging from 24 to 238 kD. Ceruloplasmin, transferrin and haptoglobin showed high levels on the seventh day after infection, with gradual increases in their concentrations until the end of the experimental period. Thus, from the data of the present study, E. stiedai is considered to be a pyogenic etiological agent for which the infection level can be monitored through the leukocyte count and serum concentrations of ceruloplasmin, transferrin and haptoglobin, and these can be recommended as complementary tests.O hemograma e proteínas de fase aguda foram avaliados durante 28 dias em coelhos adultos, machos, raça branco Nova Zelândia, infectados experimentalmente, via oral, com oocistos esporulados de Eimeria stiedai. Foram usados 50 animais distribuídos em dois grupos: grupo A infectado com 1 × 10(4 oocistos esporulados de E. stiedai e grupo B inoculado com água destilada. No 7º dia após a infecção (dpi, os animais infectados tiveram anemia, leucocitose com neutrofilia e monocitose. O método de fracionamento de proteínas por eletroforese identificou 19 proteínas de fase aguda com pesos moleculares que variaram entre 24 e 238 kD. A ceruloplasmina, transferrina e haptoglobina tiveram níveis elevados no 7° dpi com aumento progressivo de suas concentrações até o término do período experimental. Desta forma, considerando-se os dados encontrados no

  6. Aspectos clínicos e patológicos em frangos de corte (Gallus gallus domesticus infectados experimentalmente com oocistos esporulados de Eimeria acervulina Tyzzer, 1929 Clinical and pathological aspects in broilers (Gallus gallus domesticus infected experimentally with Eimeria acervulina Tyzzer, 1929 sporulated oocysts

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    Fagner Luiz da C. Freitas

    2008-03-01

    Full Text Available No presente estudo, sinais clínicos e alterações patológicas foram avaliados por 30 dias em frangos de corte, linhagem Cobb, machos, com dez dias de idade, infectados com Eimeria acervulina. Foram utilizados 192 animais distribuídos em 3 grupos: grupo A inoculado com 1x10(6 oocistos esporulados; grupo B inoculado com 1x10(5 oocistos esporulados; grupo C inoculado com água destilada. Os sinais clínicos observados foram anorexia, diarréia e apatia. As alterações patológicas macroscópicas observadas foram: enterite, hiperemia seguido de congestão intestinal, excesso de exsudato mucoso no lúmen do intestino delgado, palidez e desidratação muscular, alto acúmulo de bile na vesícula biliar e deposição de gordura hepática. A atrofia de vilosidades e alta presença de células inflamatórias foram as alterações microscópicas observadas no epitélio intestinal. Na análise histopatológica do fígado observaram-se infiltrados inflamatórios e deposição de gordura. Os resultados demonstraram que frangos de corte infectados experimentalmente com E. acervulina apresentam progressivas lesões intestinais de intensidade variável e que essas anormalidades são as principais causas de redução no desenvolvimento da ave.In the present study, clinical signs and pathological changes were evaluated during 30 days in broilers, lineage Cobb, males, with ten days old, infected with Eimeria acervulina. Were used 192 animals distributed in 3 groups: group A inoculated with 1x10(6 sporulated oocysts; group B inoculated with 1x10(5 sporulated oocysts; group C inoculated with distilled water. The clinical signs observed were anorexia, diarrhea and apathy. The pathological macroscopic changes observed were: enteritis, hyperemia with subsequent intestinal congestion, high concentration of mucus exudates in the lumen of the small intestine, pale and dehydration muscle, gall bladder full of liquid and deposition of hepatic fat. The villous atrophy

  7. Estudio anual de la eliminación de huevos y ooquistes de parásitos gastrointestinales y larvas de nemátodos pulmonares en ovinos de una estancia en Magallanes, Chile Annual study of the of egg and oocyst outputs of gastrointestinal parasites and lungworm larvae in a sheep station of Magallanes, Chile

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    G. SIEVERS

    2002-01-01

    traslado estacional de los animales sobre toda la superficie de la estanciaIn order to obtain epidemiological data of gastrointestinal and lungworm parasitosis in sheep in Magallanes, Chile, a survey was carried out at the "Entre Vientos" station during a year (September 1999 to August 2000. The tendencies of egg, oocyst and larvae outputs of parasites were determined in 3 age categories of sheep: 9.000 lambs (born in spring 1999, 1.800 ewe hoggets (1 year old and 13.500 ewes (2 or more years old, which were all kept under the traditional management system of the station. Every 14 days, faeces were obtained from the rectum of 25 animals from each age category, sampled at random, to carry out egg counts and differentiation of gastrointestinal worm larvae, counts of cestode eggs, determination of animals that were positive to oocysts of intestinal protozoa and detection of lungworm larvae. Lambs showed the greatest egg output of gastrointestinal worms during the summer, with predominance of the genus Nematodirus and Ostertagia. In ewe hoggets and ewes, the greatest output of Ostertagia and Trichostrongylus were produced in spring (November, coinciding with the temperature rise. The most contaminating category was the ewes in spring, whereas the lambs were in the summer. During spring, most of the animals were positive to Eimeria spp., but the lambs were the ones with more oocyst output. The greatest oviposition of Moniezia expansa eggs was registered in lambs and ewe hoggets during summer and spring respectively. Dictyocaulus filaria larvae were principally detected in lambs during autumn and winter. No cases of clinic parasitosis were diagnosed during the survey. The reason for this can be the extensive grazing system and the seasonal moving of the animals all over the farm surface

  8. Infecção per os de gatos com formas vegetativas de Toxoplasma gondii Nicolle & Manceaux, 1909 sem produção de oocistos Oral infections of domestic cats with vegetative forms of Toxoplasma gondii Nicolle & Manceaux, 1909 without oocyst production

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    Felippe Nery-Guimarães

    1973-01-01

    Full Text Available Quatro grupos de 6 gatos (24 gatos recém-nascidos e desmamados receberam "per os", respectivamente, suspensões de toxoplasmas de camundongos com 3-4 dias de infecção, de 4 amostras de T. gondii. Cada grupo teve um gato testemunha. Nenhum dos gatos de experiência eliminou oocistos atribuíveis a T. gondii, em períodos de observação de 6 a 20 dias; e suas fezes, conservadas 2-4 dais em bicromato de potássio a 2,5% e ministradas "per os" a camundongos, não induziram toxoplasmose nesses roedores. Com exceção dos que eram portadores de Isospora, os gatos não mostraram formas evolutivas de coccídios no epitélio intestinal. Em todos os grupos a infecção toxoplásmica foi comprovada pela positividade da reação de Sabin & Feldman (1:16 a 1:1024; e pelo isolamento de toxoplasmas pela inoculação de triturados dos seus principais órgãos em camundongos indicadores. De um modo geral, os gatos mais crescidos não mostraram sinais de doença, porém os outros, e principalmente os recém-nascidos adoeceram e vários morreram de toxoplasmose sistêmica: esplenite, hepatite, enterite, penumonia e, mais raramente, miocardite e encefalite. Os toxoplasmas foram encontrados em todos esses órgãos e, tamém, nos rins e supra-renais.Vegetative forms of 4 strains of Toxoplasma from peritoneal fluid of mice with 3-4 days of acute infection were orally administered to 4 groups of 6 newborn and wealing kittens respectivelly. Each group had a control kitten. All kittens were observed for eriods of 6-20 days ans never passed in their faeces oocysts attributabel to T. gondii. On the other hand, their faces after 2-4 days in potassium dichromate at 2.5% inoculated orally in mice didn't produce Toxoplasma infection in these animals; and with exception of the cats that have had spontaneous isosporosis (I. felis and/or I. rivolta we didn´t see evolutive forms of Coccidia in the intestinal wall of the cats. In all cat groups the toxoplasmic infection was

  9. Microbial elimination capacity of conventional water treatment for viruses, bacteria and protozoan (oo)cysts

    NARCIS (Netherlands)

    Hijnen, W.A.M.; Medema, Gerriet Jan

    2007-01-01

    In the process of Quantitative Microbial Risk Analysis prescribed by the Dutch Drinking Water Decree quantitative information about the elimination capacity of water treatment is necessary. Site specific data can be collected by monitoring the removal of surrogates like bacteriophages, indicator bac

  10. Inhibition of Plasmodium falciparum oocyst production by membrane-permeant cysteine protease inhibitor E64d.

    NARCIS (Netherlands)

    Eksi, S.; Czesny, B.; Gemert, G.J.A. van; Sauerwein, R.W.; Eling, W.M.C.; Williamson, K.C.

    2007-01-01

    During asexual intraerythrocytic growth, Plasmodium falciparum utilizes hemoglobin obtained from the host red blood cell (RBC) as a nutrient source. Papain-like cysteine proteases, falcipains 2 and 3, have been reported to be involved in hemoglobin digestion and are targets of current antimalarial d

  11. High Mobility Group Protein HMGB2 Is a Critical Regulator of Plasmodium Oocyst Development*S⃞

    OpenAIRE

    Gissot, Mathieu; Ting, Li-Min; Daly, Thomas M.; Bergman, Lawrence W.; Sinnis, Photini; Kim, Kami

    2008-01-01

    The sexual cycle of Plasmodium is required for transmission of malaria from mosquitoes to mammals, but how parasites induce the expression of genes required for the sexual stages is not known. We disrupted the Plasmodium yoelii gene encoding high mobility group nuclear factor hmgb2, which encodes a DNA-binding protein potentially implicated in transcriptional regulation of malaria gene expression. We investigated its function in vivo in the vertebrate and invertebrate ...

  12. 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...

  13. Acid-fast lipids are important structural components of oocyst walls of Cryptosporidium, Toxoplasma, and Eimeria

    Science.gov (United States)

    Coccidia are protozoan parasites that cause significant human disease and are of major agricultural importance. Cryptosporidium spp.cause diarrhea in humans and animals, while congenital Toxoplasma infections causes blindness and death. Eimeria kills chickens, so all poultry feed contain antibioti...

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

    OpenAIRE

    Srisuphanunt M.; Saksirisampant W.; Karanis P.

    2009-01-01

    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...

  15. Oocyst output and transmission rates during successive infections with Eimeria acervulina in experimental broiler flocks

    NARCIS (Netherlands)

    Velkers, F.C.; Bouma, A.; Stegeman, J.A.; Jong, de M.C.M.

    2012-01-01

    The infection dynamics of Eimeria species determine the clinical manifestation of the disease coccidiosis in poultry flocks, and a better understanding of the dynamics may contribute to improvement of control measures. Our aim was to study the course of infection and the transmission of Eimeria acer

  16. 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.

  17. A HISTORICAL PERSPECTIVE OF DETECTION METHODS FOR GIARDIA CYSTS AND CRYPTOSPORIDIUM OOCYSTS IN WATER

    Science.gov (United States)

    In the mid-20th century Giardia was classified as a non-pathogenic commensal organism and Cryptosporidium was not recognized yet. However as early as 1946 a waterborne outbreak of giardiasis was suspected. From 1965 to 1979 it became clear that Giardia lamblia was indeed a human ...

  18. Viability staining of Cryptosporidium oocysts and Giardia cysts combined with flow cytometry

    NARCIS (Netherlands)

    Schets FM; Medema GJ; MGB

    1998-01-01

    Flow cytometrie als extra zuiveringsstap heeft de detectiemethode voor Cryptosporidium oocysten en Giardia cysten in water verbeterd. Door middel van flow cytometrie kunnen (oo)cysten gescheiden worden van het meeste debris dat in waterconcentraten aanwezig is, waardoor de toepassing van fluorogene

  19. Oocyst output and transmission rates during successive infections with Eimeria acervulina in experimental broiler flocks

    NARCIS (Netherlands)

    Velkers, F.C.; Bouma, A.; Stegeman, J.A.; Jong, de M.C.M.

    2012-01-01

    The infection dynamics of Eimeria species determine the clinical manifestation of the disease coccidiosis in poultry flocks, and a better understanding of the dynamics may contribute to improvement of control measures. Our aim was to study the course of infection and the transmission of Eimeria acer

  20. EFFECT OF LACTOBACILLUS AND BIFIDOBACTERIUM ON CRYPTOSPORIDIUM PARVUM OOCYST VIABILITY. (R826138)

    Science.gov (United States)

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

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

    Science.gov (United States)

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

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

    Science.gov (United States)

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

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

    Science.gov (United States)

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

  4. INACTIVATION OF CRYPTOSPORIDIUM OOCYSTS IN A PILOT-SCALE BUBBLE-DIFFUSER CONTACTOR II: MODEL VALIDATION AND APPLICATION. (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...

  5. Factors affecting recovery efficiency in isolation of Cryptosporidium oocysts and Giardia cysts from vegetables for standard method development.

    Science.gov (United States)

    Robertson, L J; Gjerde, B

    2001-11-01

    While recently published techniques for recovering parasites from fruits and vegetables demonstrate a marked increase in efficiency and utility, there is still scope for further improvement in developing a standard method, particularly with difficult, but important, sample matrices such as bean sprouts. Herein, a number of parameters used in published techniques are investigated more closely. While sample size reduction may improve recovery efficiency because of a range of factors, it is important to keep the sample large enough for detection of low-level contamination. Age of sample is also important, and samples should be as fresh as possible. Elution procedures may contribute to losses of Giardia and should be more thoroughly investigated. Improved immunomagnetic separation techniques currently coming onto the market also have the potential to increase recovery efficiency substantially, even with difficult samples such as aged bean sprouts. However, merely increasing magnetic strength of the capturing magnet does not affect recovery efficiency, which must be reliant on a superior bead system, buffering system, or both.

  6. The Effect of Anthelmintic Treatment on Coccidia Oocyst Shedding in a Wild Mammal Host with Intermittent Cestode Infection

    Directory of Open Access Journals (Sweden)

    Radovan Václav

    2014-01-01

    Full Text Available While hosts are routinely exploited by a community of parasite species, the principles governing host responses towards parasites are unclear. Identifying the health outcomes of coinfections involving helminth macroparasites and microparasites is one area of importance for public and domestic animal health. For instance, it is controversial how deworming programmes affect incidence and severity of such important microparasite diseases as malaria. One problem is that most study systems involve domestic and laboratory animals with conditions hardly comparable to those of free-living animals. Here, we study the effect of anthelmintic treatment on coccidia infection intensity in wild Alpine marmots, M. marmota. Our results lend support to the hypothesis that helminth infection has a positive effect on concurrent microparasite infection. However, our work also points to the fact that within-host interactions between helminths and microparasites are context-dependent and can turn to negative ones once helminth burdens increase. Our study suggests that coccidia benefit from intermittent helminth infection in marmots due to the protective effects of helminth infection only during the early phase of the host’s active season. Also, the marmot’s response towards coccidia infection appears optimal only under no helminth infection when the host immune response towards coccidia would not be compromised, thereby pointing to the importance of regular intestinal helminth elimination by marmots just before hibernation.

  7. 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...

  8. The effects of combining Artemisia annua and Curcuma longa ethanolic extracts in broilers challenged with infective oocysts of Eimeria acervulina and E. maxima

    Science.gov (United States)

    Due to an increasing demand for natural products to control coccidiosis in broilers we investigated the effects of supplementing a combination of ethanolic extracts of Artemisia annua and Curcuma longa in drinking water. Three different dosages of this herbal mixture were compared with a negative co...

  9. 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

    Due to an increasing demand for natural products to control coccidiosis in broilers, we investigated the effects of supplementing a combination of ethanolic extracts of Artemisia annua and Curcuma longa in drinking water. Three different dosages of this herbal mixture were compared with a negativ...

  10. 离体条件下球虫卵囊子孢子囊的释放%Studies on Sporocyst Releasing from Oocyst of Eimeria tenella in Vitro

    Institute of Scientific and Technical Information of China (English)

    安健; 王黎霞; 汪明; 张建军; 李翠莲; 高立娜

    2006-01-01

    利用柔嫩艾美耳球虫(Eimeria tenella)为试验虫株,盖玻片、载玻片为实验材料,用拇指适当施加压力时,在400倍的光学显微镜下,观察了柔嫩艾美耳球虫卵囊破裂和子孢子囊释放的过程,结果发现,卵囊在适当压力的作用下,在卵囊一端发生破裂,随后子孢子囊逐个从破裂口处溢出,溢出后的子孢子囊内含有2个子孢子,说明球虫卵囊虽然对常规的消毒剂抵抗力很强,但是在鸡的消化道内,尤其在肌胃内球虫卵囊很容易破裂而释放子孢子囊.

  11. INTERFERON-GAMMA EXPRESSION IN JEJUNAL BIOPSIES IN EXPERIMENTAL HUMAN CRYPTOSPORIDIOSIS CORRELATES WITH PRIOR SENSITIZATION AND CONTROL OF OOCYST EXCRETION. (R824759,R828035)

    Science.gov (United States)

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

  12. Removal of Fecal Indicators, Pathogenic Bacteria, Adenovirus, Cryptosporidium and Giardia (oo)cysts in Waste Stabilization Ponds in Northern and Eastern Australia.

    Science.gov (United States)

    Sheludchenko, Maxim; Padovan, Anna; Katouli, Mohammad; Stratton, Helen

    2016-01-02

    Maturation ponds are used in rural and regional areas in Australia to remove the microbial loads of sewage wastewater, however, they have not been studied intensively until present. Using a combination of culture-based methods and quantitative real-time PCR, we assessed microbial removal rates in maturation ponds at four waste stabilization ponds (WSP) with (n = 1) and without (n = 3) baffles in rural and remote communities in Australia. Concentrations of total coliforms, E. coli, enterococci, Campylobacter spp., Salmonella spp., F+ RNA coliphage, adenovirus, Cryptosporidium spp. and Giardia (oo) cysts in maturation ponds were measured at the inlet and outlet. Only the baffled pond demonstrated a significant removal of most of the pathogens tested and therefore was subjected to further study by analyzing E. coli and enterococci concentrations at six points along the baffles over five sampling rounds. Using culture-based methods, we found a decrease in the number of E. coli and enterococci from the initial values of 100,000 CFU per 100 mL in the inlet samples to approximately 1000 CFU per 100 mL in the outlet samples for both bacterial groups. Giardia cysts removal was relatively higher than fecal indicators reduction possibly due to sedimentation.

  13. Removal of sulphite-reducing clostridia spores by full-scale water treatment processes as a surrogate for protozoan (oo)cysts removal

    NARCIS (Netherlands)

    Hijnen, W.A.M.; Willemsen-Zwaagstra, J.; Hiemstra, P.; Medema, Gerriet Jan; Kooij, D. van der

    2000-01-01

    At eight full-scale water treatment plants in the Netherlands the removal of spores of sulphite-reducing clostridia (SSRC) was determined. By sampling and processing large volumes of water (1 up to 500 litres) SSRC were detected after each stage of the treatment. This enabled the assessment of the

  14. INTERFERON-GAMMA EXPRESSION IN JEJUNAL BIOPSIES IN EXPERIMENTAL HUMAN CRYPTOSPORIDIOSIS CORRELATES WITH PRIOR SENSITIZATION AND CONTROL OF OOCYST EXCRETION. (R824759,R828035)

    Science.gov (United States)

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

  15. 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.

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

    Science.gov (United States)

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

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

    Science.gov (United States)

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

  18. Dicty_cDB: SLJ419 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available |CB367289.1 E38 early-oocyst library Plasmodium berghei/Anopheles stephensi mixe...7 |CB367287.1 E830 early-oocyst library Plasmodium berghei/Anopheles stephensi mixed EST library cDNA clone ...oocyst library Plasmodium berghei/Anopheles stephensi mixed EST library cDNA clon...ly-oocyst library Plasmodium berghei/Anopheles stephensi mixed EST library cDNA clone E3015 similar to Plasm

  19. Development of method for detecting Cyclospora oocysts on the surface of fresh fruits and vegetables%水果及蔬菜表面环孢子虫卵囊检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    孙芳芳; 罗楠楠; 李俊强; 赵金凤; 李廷文; 黄建营; 张龙现

    2014-01-01

    为了建立检测果蔬表面人源卡耶坦环孢子虫(Cyclospora cayetanensis)卵囊的方法,分别用50,500个环孢子虫卵囊对环孢子虫卵囊阴性的新鲜果蔬进行人工污染,并设置阴性对照;使用含有0.1% Alconox@,0.1%雕牌,0.1%立白、0.1%白猫的4种洗涤液和去离子水,在摇床中洗脱样品;然后,反复离心3次收集卵囊并提取卵囊DNA;最后,基于环孢子虫18S rRNA基因住点对卵囊DNA样品进行巢式PCR扩增.结果表明,卵囊污染吸附最佳时间为24 h,样品洗脱最佳时间为30 min.其中,只用去离子水时,生菜、西葫芦、黄瓜、西瓜、甜瓜和草莓的检测敏感性可达到50个卵囊,而西红柿可达到500个卵囊;当用不同洗涤剂时,各样品的检测敏感性均可达到50个卵囊.不同洗涤剂之间检测敏感性无显著差异.该研究初步建立检测水果、蔬菜污染环孢子虫卵囊的技术,采用此方法可特异、敏感、快速的检测果蔬表面环孢子虫卵囊,为食源性环孢子虫病暴发溯源提供技术支持.

  20. Salivary IgA against sporozoite-specific embryogenesis-related protein (TgERP) in the study of horizontally transmitted toxoplasmosis via T. gondii oocysts in endemic settings

    Science.gov (United States)

    The prevalence of toxoplasmosis was investigated in endemic settings in Brazil, and calculated by measuring antibodies in two ELISA systems: 1) IgG and IgM from sera tested by commercial conventional ELISA, and 2) IgA, from saliva, and IgG from sera samples tested against a sporozoite-specific prote...

  1. A new set of primers directed to 18S rRNA gene for molecular identification of Cryptosporidium spp. and their performance in the detection and differentiation of oocysts shed by synanthropic rodents.

    Science.gov (United States)

    Silva, Sheila O S; Richtzenhain, Leonardo J; Barros, Iracema N; Gomes, Alessandra M M C; Silva, Aristeu V; Kozerski, Noemila D; de Araújo Ceranto, Jaqueline B; Keid, Lara B; Soares, Rodrigo M

    2013-11-01

    Cryptosporidium spp. are cosmopolitan protozoa that infect fishes, reptiles, amphibians, birds and mammals. More than 20 species are recognized within this genus. Rodents are a group of abundant and ubiquitous organisms that have been considered reservoirs of Cryptosporidium for humans and livestock. The aim of this study was to design specific primers for the gene encoding 18S rRNA, potentially capable of amplifying any species or genotype of Cryptosporidium spp. and evaluate the diagnostic attributes of the nested-PCR based on such probes. The primers were designed to amplify the shortest segment as possible to maximize the sensitivity of the test, but preserving the discriminatory potential of the amplified sequences for phylogenetic inferences. The nested-PCR standardized in this study (nPCR-SH) was compared in terms of sensitivity with another similar assay (nPCR-XIAO) that has been largely used for the detection and identification of Cryptosporidium spp. worldwide. We also aimed to molecularly characterize samples of Cryptosporidum spp. isolated from synanthropic rodents using these probes. Forty-five rodents were captured in urban areas of the municipality of Umuarama, Paraná State, Brazil. Fecal samples were submitted to three molecular tests (nested-PCRs), two of them targeted to the 18S rDNA gene (nPCR-SH and nPCR-XIAO) and the third targeted to the gene encoding actin (nPCR-actin). The nPCR-SH was tested positive on samples of Cryptosporidum parvum, Cryptosporidum andersoni, Cryptosporidum meleagridis, Cryptosporidum hominis, Cryptosporidum canis, and Cryptosporidum serpentis. Sixteen samples of rodents were positive by nPCR-SH, six by nPCR-XIAO and five by nPCR-actin. Sequencing of amplified fragments allowed the identification of Cryptosporidum muris in three samples of Rattus rattus, and two genotypes of Cryptosporidium, the genotypes mouse II and III. Cryptosporidium genotype mouse II was found in one sample of Mus musculus and genotype mouse III, in twelve samples, being five from R. rattus and seven from M. musculus. The results of this study demonstrated that the primers designed for detection of Cryptosporidium spp. were more efficient than those used in the nPCR-XIAO. Genotypes or species of Cryptosporidium that can be usually transmitted for human beings and livestock were not found in synanthropic rodents, suggesting that the importance of these animals in zoonotic transmission of cryptosporidiosis should be revisited.

  2. 艾美耳属球虫卵囊基因组DNA不同提取方法的比较%Comparison of the Extraction of Eimeria Oocyst Genomic DNA

    Institute of Scientific and Technical Information of China (English)

    李继东

    2009-01-01

    为了获得高质量的艾美耳属(Eimeria)球虫卵囊基因组DNA,采用了4种不同方法提取3种艾美耳属球虫卵囊的基因组DNA.对比结果表明,采用机械法破裂球虫卵囊壁,用裂解液裂解孢子囊和子袍子,释放出DNA,在保持碱性的环境下(pH=8.0),以苯酚-氯仿-异戊醇抽提,乙醇沉淀,可获得高质量的DNA分子,经PCR检验为艾美耳属球虫卵囊基因组DNA.

  3. ENVIRONMENTAL TECHNOLOGY VERIFICATION REPORT PHYSICAL REMOVAL OF CRYPTOSPORIDIUM OOCYSTS AND GIARDIA CYSTS IN DRINKING WATER PALL CORPORATION WPM-1 MICROFILTRATION SYSTEM AT PITTSBURGH, PA - NSF 00/09/EPADW395

    Science.gov (United States)

    Verification testing of the Pall Corporation WPM-1 Microfiltration Pilot System was conducted from February 3 to March 5, 1999. The Pall WPM-1 membrane is a hollow fiber type microfiltration membrane made of polyvinylidenefluoride (PVDF). It has a 0.1 micrometer ( m) nominal por...

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

    Science.gov (United States)

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

    2012-05-01

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

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

    Science.gov (United States)

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

    2015-12-01

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

  6. 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.

  7. Free-living amoebae as vectors of cryptosporidia.

    Science.gov (United States)

    Scheid, Patrick L; Schwarzenberger, Rafael

    2011-08-01

    In the present article, the study to examine the ability of free-living amoebae (FLA) to serve as vectors of cryptosporidia is presented. Ten strains of different free-living amoebae of the FLA collection of the Parasitology Lab at Koblenz were cultivated in the presence of Cryptosporidium parvum oocysts. After phagocytosis and ingestion, the oocysts could be found in food vacuoles within the cytoplasm of the trophozoites of two different FLA strains. The uptake and the transport of the oocysts within the trophozoites could be demonstrated in an Acanthamoeba sp. (group II) strain (maximum, three oocysts; average, one oocyst) as well as in a Thecamoeba quadrilineata strain (maximum, 15 oocysts; average, eight oocysts), with the help of light microscopy. We found that these free-living amoebae can temporarily harbour cryptosporidia, thus supporting the suggestion that FLA may act as carriers and vehicles for cryptosporidia. However, proliferation did not take place within the host amoebae. No cryptosporidium oocysts were found within the cysts of the amoebae. To our knowledge, this is the first study to determine the "host range" of free-living amoebae as vectors and vehicles of cryptosporidia. Free-living amoebae appear able to act as carriers or vectors of the oocysts and thus may play a certain role in the transmission of cryptosporidia.

  8. Comparison of Eimeria species distribution and salinomycin resistance in commercial broiler operations utilizing different coccidiosis control strategies.

    Science.gov (United States)

    Jenkins, M; Klopp, S; Ritter, D; Miska, K; Fetterer, R

    2010-09-01

    The purpose of the present study was to evaluate the species composition and salinomycin sensitivity of Eimeria oocysts isolated from commercial broiler farms that differed by means of coccidiosis control (anticoccidial drugs [ACD] vs. live oocyst vaccines [VAC]). A comparison of Eimeria species composition and salinomycin sensitivity was also made before and after a producer switched from salinomycin to live oocyst vaccines. In general, no significant difference was observed in the concentration of Eimeria spp. oocysts in litter from VAC-utilizing farms compared to litter from ACD-utilizing farms. Application of PCR-based methods to detect coccidia found that Eimeria species distribution in litter from VAC operations more closely resembled the species composition in the live oocyst vaccines. Drug sensitivity testing found that Eimeria oocysts from VAC operations displayed greater salinomycin sensitivity as measured by weight gain and feed conversion efficiency compared to oocysts from ACD farms. These findings provide additional evidence for the usefulness of live oocyst vaccines to restore ionophore sensitivity in poultry operations that contain an ionophore-resistant population of Eimeria spp. oocysts.

  9. Experimental Infection of Swine by Isospora suis Biester 1934 for Species Confirmation

    Directory of Open Access Journals (Sweden)

    Silvia Maria Oliveira Sayd

    1998-11-01

    Full Text Available A survey of Isospora suis performed in 177 faecal samples from 30 swine farms detected thin wall type I. suis oocysts in seven samples. This type of oocyst measuring 23.9 by 20.7 mm had a retracted thin wall similar to that of the genus Sarcocystis. This type of oocysts, isolated from four different faecal samples, was inoculated in four-five-days-old piglets free of contamination in order to verify the life cycle and pathogenicity of the species. The pigs were kept in individual metal cages and fed with cow milk. Daily faecal collections and examinations were performed until the 21st day after infection. MacMaster and Sheather' s methods were used for oocyst counting and identification. Infected piglets produced yellowish-pasty diarrhoea with slight dehydration. The prepatent and patent periods were respectively from 6 to 9 and 3 to 10 days after infection. Oocyst elimination was interrupted on the 10th and 11th days after infection with biphasic cycles. Thin and thick wall oocysts were detected in the same faecal samples. Thin walls were not observed in unsporulated oocysts. The observations suggest that this type of oocysts could appear in specific strains which occur in the later stages of their development. These oocysts seem to be responsible for clinical and pathogenic signs of neonatal isosporosis in pigs.

  10. Direct and indirect immunosuppression by a malaria parasite in its mosquito vector

    NARCIS (Netherlands)

    Boëte, C.H.J.J.; Paul, R.E.L.; Koëlla, J.C.

    2004-01-01

    Malaria parasites develop as oocysts within the haemocoel of their mosquito vector during a period that is longer than the average lifespan of many of their vectors. How can they escape from the mosquito's immune responses during their long development? Whereas older oocysts might camouflage themsel

  11. First isolation of Hammondia hammondi from a cat from Ethiopia

    Science.gov (United States)

    Toxoplasma gondii oocysts are morphologically and antigenically similar to oocysts of another feline coccidian, Hammondia hammondi. The distinction between H. hammondi and T. gondii is important from an epidemiological perspective because all isolates of T. gondii are potentially pathogenic for hum...

  12. Protection of non-immunized broiler chicks housed with immunized cohorts against infection with Eimeria maxima and E. acervulina

    Science.gov (United States)

    The use of live oocyst vaccines is becoming increasingly important in the control of avian coccidosis in broiler chicks. Knowledge of the mechanisms of how chicks uptake oocysts and become immune is important for optimizing delivery of live vaccines. The current study tests the hypothesis that chick...

  13. F-actin distribution and function during sexual development in Eimeria maxima.

    Science.gov (United States)

    Frölich, Sonja; Wallach, Michael

    2015-06-01

    To determine the involvement of the actin cytoskeleton in macrogametocyte growth and oocyst wall formation, freshly purified macrogametocytes and oocysts were stained with Oregon Green 514 conjugated phalloidin to visualize F-actin microfilaments, while Evans blue staining was used to detect type 1 wall forming bodies (WFB1s) and the outer oocyst wall. The double-labelled parasites were then analysed at various stages of sexual development using three-dimensional confocal microscopy. The results showed F-actin filaments were distributed throughout the entire cytoplasm of mature Eimeria maxima macrogametocytes forming a web-like meshwork of actin filaments linking the type 1 WFBs together into structures resembling 'beads on a string'. At the early stages of oocyst wall formation, F-actin localization changed in alignment with the egg-shaped morphology of the forming oocysts with F-actin microfilaments making direct contact with the WFB1s. In tissue oocysts, the labelled actin cytoskeleton was situated underneath the forming outer layer of the oocyst wall. Treatment of macrogametocytes in vitro with the actin depolymerizing agents, Cytochalasin D and Latrunculin, led to a reduction in the numbers of mature WFB1s in the cytoplasm of the developing macrogametocytes, indicating that the actin plays an important role in WFB1 transport and oocyst wall formation in E. maxima.

  14. Sericea Lespedeza has no anticoccidial effect when included in the diet of chickens infected with three species of Eimeria.

    Science.gov (United States)

    Rathinam, T; Gadde, U; Chapman, H D

    2014-05-28

    Anticoccidial effects of Sericea Lespedeza (SL) included in the diet at different levels were evaluated in chickens following oral infection with sporulated oocysts of either Eimeria acervulina, Eimeria maxima or Eimeria tenella. A series of experiments were conducted to determine the effect of SL upon the ability of the parasites to multiply in the intestine, and the effect on bodyweight gain, feed intake, and feed conversion following infection. Chicks infected with a low dose of oocysts (500 oocysts/bird) of E. acervulina or E. maxima did not show differences in the numbers of oocysts produced in the feces whether they were given 0, 1, 2, or 4% SL in the diet. There was no significant difference in the weight gain, feed intake, or FCR of birds infected with high doses of E. acervulina or E. maxima (200,000 or 100,000 oocysts/bird respectively) whether 0, 1, 2, or 4% SL was included in the feed. No significant difference in the numbers of oocysts in the feces, weight gain, feed intake, and FCR of birds infected with E. tenella (low dose of 500 oocysts; high dose of 50,000 oocysts per bird) whether 4% SL was included in the feed. The results of this study indicate that SL has no anticoccidial activity against Eimeria species in the chicken.

  15. 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

  16. Toxoplasmosis in sentinel chickens (Gallus domesticus) in New England farms: seroconversion, distribution of tissue cysts in brain, heart, and skeletal muscle by bioassay in mice and cats

    Science.gov (United States)

    Free-range chickens are a good indicator of soil contamination with oocysts because they feed from the ground and they are also an important source of infection for cats that in turn shed oocysts after eating tissues of intermediate hosts. Little is known of the epidemiology of toxoplasmosis in chic...

  17. Direct and indirect immunosuppression by a malaria parasite in its mosquito vector

    NARCIS (Netherlands)

    Boëte, C.H.J.J.; Paul, R.E.L.; Koëlla, J.C.

    2004-01-01

    Malaria parasites develop as oocysts within the haemocoel of their mosquito vector during a period that is longer than the average lifespan of many of their vectors. How can they escape from the mosquito's immune responses during their long development? Whereas older oocysts might camouflage

  18. Four new species of coccidia (Apicomplexa: Eimeriidae) from Owen Stanley Skinks, Papuascincus stanleyanus (Sauria: Scincidae), from Papua New Guinea

    Science.gov (United States)

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

    2014-01-01

    Between September and November 1991, 12 Owen Stanley skinks, Papuascincus stanleyanus (Booulenger) were collected from various localities on Papua New Guinea and examined for coccidians. Six (50%) were found to harbour four eimerians that we describe here as new. Oocysts of Eimeria burseyi sp. n. were elongate to ellipsoidal with a bilayered wall and measured (length x width, L x W) 36.0 x 24.0 microm, with a L/W ratio of 1.5. Both micropyle and oocyst residuum were absent, but a polar granule was present. Oocysts of Eimeria goldbergi sp. n. were ellipsoidal, with a bilayered wall, and measured 21.4 x 16.1 microm; L/W ratio was 1.3. Both micropyle and oocyst residuum were absent, but a single or fragmented polar granule was present. Oocysts of Eimeria boulengeri sp. n. were spheroidal to slightly subspheroidal, with a thin, single-layered wall that readily collapses, and measured 16.0 microm, L/W ratio was 1.0. Both micropyle and oocyst residuum were absent, but usually one (sometimes two) polar granule(s) were present. Oocysts of Eimeria niuginiensis sp. n. were oblong to tapered with a bilayered wall, and measured 20.0 x 13.1 microm; L/W ratio was 1.5. A micropyle, oocyst residuum and polar granule were absent. To our knowledge, these represent the only coccidians ever described from P. stanleyanus.

  19. 盐城地区山羊球虫感染情况调查及单种卵囊感染试验%Investigation on coccidia of Eimeria arloingi and E. ninakohlyakimovca in goats in Yancheng district and their infection with oocysts

    Institute of Scientific and Technical Information of China (English)

    戴亚斌; 陶红; 符敖齐; 陶建平

    1996-01-01

    @@ 球虫病是山羊多见的一种原虫性疾病,常引起山羊拉稀、消瘦、衰弱,对羔羊的危害尤甚,可使其生长发育停滞,生产性能下降,甚至造成死亡.我们于1994年对盐城地区山羊球虫感染情况进行了调查,并对Eimeria arloingi和E.ninakohlyakimovae分别进行了单种卵囊感染试验.

  20. Study on geographical strains of Eimeria maxima Ⅳ.Analysis on antigens and SDS-PAGE of sporulated oocysts proteins%巨型艾美耳球虫(Eimeria maxima)地理株研究Ⅳ.孢子化卵囊蛋白的SDS-PAGE与抗原性分析

    Institute of Scientific and Technical Information of China (English)

    李文超; 陶建平; 顾有方; 沈永林

    2010-01-01

    利用聚丙烯酰胺凝胶电泳技术(SDS-PAGE)分析比较了11株巨型艾美耳球虫(Eimeria maxima)孢子化卵囊可溶性蛋白.用上海株、扬州株和美国株的阳性血清,分别对上海株、扬州株和美国株的孢子化卵囊可溶性蛋白进行Western blotting分析,比较这3株球虫孢子化卵囊可溶性蛋白的抗原性.结果显示,11株巨型艾美耳球虫孢子化卵囊可溶性蛋白的SDS-PAGE电泳图谱相似,至少有18条电泳条带,其中有7条相对明显的主带,相对分子质量分别为100 200、77 000、63 200、48 300、41 200、36 300和30 000.Western blotting分析显示,在同源株间与异源株问的印迹图谱未见差异,均有4条免疫印迹务带,相对分子质量分别为103 700、64 200、43 400和37 100.

  1. 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.

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

    Science.gov (United States)

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

    2008-02-01

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

  3. Toltrazuril (Baycox) treatment against coccidiosis caused by Eimeria sp. in Japanese quails (Coturnix coturnix japonica).

    Science.gov (United States)

    Sokół, R; Gesek, M; Raś-Noryńska, M; Michalczyk, M

    2014-01-01

    Coccidiosis is the most predominant parasitic disease affecting Japanese quails (Coturnix coturnix japonica) in commercial farms. Coccidiosis as a subclinical infection is difficult to diagnose without parasitological examinations. Oocysts of two Eimeria species, E. bateri and E. tsunodai, were determined in the analysed quail flock. Infected birds were administered Baycox 2.5% at the dose of: group I--7 mg toltrazuril/kg BW per day provided in drinking water (1.5 ml/0.5 1 H2O) that was available 24 h for 2 days, group II--14 mg/kg BW (3 ml/0.5 1 H2O), and group III-- 24.5 mg/kg BW (5 ml/0.5 1 H2O); in groups II and III, the solutions were available 8 h/24 h for 2 days. After the first day of the treatment, the number of excreted oocysts (OPG - oocysts per gram) increased, a steady decrease in oocyst counts began on the second day of Baycox administration and lasted until a three-day period when no oocyst were determined in faecal samples. Regardless of the dose applied, toltrazuril (Baycox) completely eliminated E. bateri coccidia and led to a highly significant reduction in the number of E. tsunodai oocysts. The results suggest that the effectiveness of toltrazuril varies depending on coccidia species and developmental stages of the parasite. From the clinical point of view, the treatment applied significantly reduces the number of coccidia oocysts in commercial flocks of Japanese quails.

  4. Effects of artemisinin in broiler chickens challenged with Eimeria acervulina, E. maxima and E. tenella in battery trials.

    Science.gov (United States)

    Pop, Loredana; Györke, Adriana; Tǎbǎran, Alexandru Flaviu; Dumitrache, Mirabela Oana; Kalmár, Zsuzsa; Magdaş, Cristian; Mircean, Viorica; Zagon, Diana; Balea, Anamaria; Cozma, Vasile

    2015-12-15

    Four experiments were conceived in order to test the efficacy of artemisinin, a sesquiterpene lactone derived from Artemisia annua, in single experimental infection of broiler chickens with Eimeria acervulina (1 × 10(5) oocysts), Eimeria maxima (5 × 10(4) oocysts) or Eimeria tenella (1 × 10(4) oocysts), and mixed infection with all 3 species (3.2 × 10(4) Eimeria spp. oocysts). For each experiment, three different dosages of artemisinin (5, 50 and 500 ppm) were compared with a negative control (uninfected, unmedicated), a positive control (infected, unmedicated) and a classical anticoccidial (monensin). The weight gain (WG), feed conversion ratio (FCR), oocysts shedded per gram of feces (OPG), lesion score, oocysts sporulation rates and mortality rate were recorded in all groups. The dosage of 5 ppm of artemisinin improved the WG and FCR for the chickens infected with E. acervulina. The OPG was significantly decreased in all the groups medicated with artemisinin and challenged with a mixed infection (p ≤ 0.01). The lesion score of the chickens challenged with Eimeria was reduced by different concentrations of artemisinin, depending on the species involved, but this compound did not have a positive effect on the lesions caused by E. acervulina. Histopathological analysis revealed superficial erosions of the intestinal mucosa, mixt. mononuclear and heterophilic inflammatory infiltrate in the lamina propria and intralesional presence of various developmental stages of parasite in groups infected with Eimeria spp.The sporulation rate of E. acervulina and E. maxima oocysts was significantly affected by 500 ppm of artemisinin, whilst the dosage of 5 ppm affected the sporulation of E. tenella oocysts. These data suggest that artemisinin is not effective against single eimerian infections but could be used as an alternative in mixed coccidiosis, especially if its effect on the oocysts sporulation would be fully investigated.

  5. Coccidia in passerines from the Nevado de Toluca National Park, Mexico.

    Science.gov (United States)

    Medina, Juan P; Salgado-Miranda, Celene; García-Conejo, Michele; Galindo-Sánchez, Karla P; Mejía-García, Cristian J; Janczur, Mariusz K; Gomes Lopes, Carlos W; Berto, Bruno P; Soriano-Vargas, Edgardo

    2014-03-01

    In this study, we found unsporulated coccidia oocysts in passerines from the Nevado de Toluca National Park, Mexico. We captured birds and took samples of their droppings during three field visits. We examined a total of 72 fecal samples and found unsporulated coccidia oocysts in 10 samples from five passerine species: Atlapetes pileatus (3), Cardelina ruber (1), Mniotilta varia (1), Oreothlypis celata (2) and Regulus calendula (3). This appears to be the first recorded study of unsporulated coccidia oocysts in passerine species from Mexico.

  6. Cryptosporidium infections in children in Durban

    African Journals Online (AJOL)

    1991-03-16

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

  7. Australian dingoes are definitive hosts of Neospora caninum.

    Science.gov (United States)

    King, Jessica S; Slapeta, Jan; Jenkins, David J; Al-Qassab, Sarwat E; Ellis, John T; Windsor, Peter A

    2010-07-01

    To provide objective data on the potential role of dingoes (Canis lupus dingo) in the life cycle of Neospora caninum in Australia, the production of N. caninum oocysts by experimentally infected canids was investigated. Three dingo pups raised in captivity and three domestic dogs were fed tissue from calves infected with an Australian isolate of N. caninum, Nc-Nowra. Oocysts of N. caninum, confirmed by species-specific PCR, were shed in low numbers by one dingo pup at 12-14 days p.i. The remaining animals did not shed oocysts. Furthermore, the blood from two out of three dingoes tested positive for DNA of N. caninum using PCR tests at 14 and 28 days p.i. Oocyst shedding from the intestinal tract of a dingo demonstrates that dingoes are definitive hosts of N. caninum and horizontal transmission of N. caninum from dingoes to farm animals and wildlife may occur in Australia.

  8. The sylvatic cycle of Neospora caninum: where do we go from here?

    Science.gov (United States)

    Rosypal, Alexa C; Lindsay, David S

    2005-10-01

    Bovine abortions due to Neospora caninum infection are a major cattle-production problem worldwide. The parasite is readily maintained in cattle populations by vertical transmission. The domestic dog excretes oocysts in its feces and, after sporulation, these oocysts are infectious to cattle. Current control measures are aimed at culling infected cows and limiting the access of cattle to infective oocysts. The recent revelations that coyotes (Canis latrans) can excrete N. caninum oocysts in their feces and that white-tailed deer (Odocoileus virginianus) are natural intermediate hosts of the parasite demonstrate the existence of a sylvatic cycle of neosporosis in North America. This complicates parasite-prevention programs but opens many new and exciting avenues of research. Similar canid-ruminant sylvatic cycles might exist in other countries and, if so, need to be investigated.

  9. Deformability Assessment of Waterborne Protozoa Using a Microfluidic-Enabled Force Microscopy Probe

    NARCIS (Netherlands)

    McGrath, John S.; Quist, Jos; Seddon, James R.T.; Lai, Stanley C.S.; Lemay, Serge G.; 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 Cryptosporidiu

  10. New and emended descriptions of gregarines from flour beetles (Tribolium spp. and Palorus subdepressus: Coleoptera, Tenebrionidae).

    Science.gov (United States)

    Janovy, J; Detwiler, J; Schwank, S; Bolek, M G; Knipes, A K; Langford, G J

    2007-10-01

    The following new gregarine taxa are described from larvae of flour beetles (Coleoptera: Tenebrionidae): Awrygregarina billmani, n. gen., n. sp., from Tribolium brevicornis; Gregarina cloptoni, n. sp., from Tribolium freemani; Gregarina confusa, n. sp., from Tribolilum confusum; and Gregarina palori, n. sp., from Palorus subdepressus. In addition, the description of Gregarina minuta Ishii, 1914, from Tribolium castaneum, is emended. Scanning electron micrograph studies of these species' oocysts reveal differences in surface architecture. The Gregarina species have oocysts with longitudinal ridges, visible with SEM, whereas Awrygregarina billmani oocysts have fine circumferential striations; surface architecture is the main feature distinguishing the 2 gregarine genera. Although parasites from adult beetles are not included in the descriptions, adults of all host species can be infected experimentally using oocysts from the new taxa.

  11. Gregarina triboliorum (Eugregarinida: Gregarinidae) n. sp. from Tribolium confusum and resolution of the confused taxonomic history of Gregarina minuta Ishii, 1914.

    Science.gov (United States)

    Watwood, S; Janovy, J; Peterson, E; Addison, M A

    1997-06-01

    The septate gregarine parasites of flour beetles (Tribolium spp.) include Gregarina minuta Ishii, 1914, a relatively small species in which both primite and satellite possess an obvious protomerite, and a larger species that lacks the satellite protomerite. The latter species has been placed in the genera Didymophyes and Hirmocystis by various authors, but studies reported here demonstrate that this species, herein described as Gregarina triboliorum, exhibits early pairing and produces oocyst chains, both characteristics of the genus Gregarina. The oocysts of this new species are described for the first time. In addition, experimental infections using oocyst from single gametocysts reveal that oocyst chain number is variable but is typically 1, 2, or 4. Prior experiments involving a related beetle, Tenebrio molitor, demonstrated extreme host specificity within the 4 Gregarina species parasitizing larval and adult hosts. However, G. triboliorum is not limited either stadially or specially, infecting both adults and larvae of Tribolium confusum and Tribolium castaneum.

  12. [Investigation of the prevalence of gastrointestinal parasites in aviary birds in homes in the Van province].

    Science.gov (United States)

    Gül, Abdurrahman; Ciçek, Mutalip

    2009-01-01

    This study was conducted to investigate the prevalence of gastrointestinal parasites in aviary birds in the Van province. For this purpose, fecal samples were collected from aviary birds (6 parrots, 5 nightingales, 24 canaries and 35 lovebirds) in Van. In the parasitology laboratory, seventy fecal samples were examined using the native and sedimentation technique for helminth eggs, Eimeria sp. oocysts and Giardia sp. cysts. The same samples were examined by modified-acid-fast staining for Cryptosporidium sp. oocysts. Prevalence of gastrointestinal parasites was found to be 8.57% (6/70) in Van. Eimeria sp. oocysts were detected in lovebird (8.57%) and canaries (8.33%). Cryptosporidium sp. oocysts were detected only in parrots (16.67%). Helminth eggs and Giardia sp. cysts were not found in aviary birds. However no parasites were detected in nightingales.

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

    Science.gov (United States)

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

    2014-01-01

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

  14. Prevalence of Parasitic Contamination of Salad Vegetables in Ilorin ...

    African Journals Online (AJOL)

    EBUBE AMAECHICHARLES

    parasitic contamination of food product varies from one area to another. ... oocysts to adverse temperatures, dessication, natural irradiation, chemicals and ..... cestodes is due to the inadequate provision of toilet facilities in Ipata area, ...

  15. Reclassification of Eimeria pogonae Walden (2009) as Choleoeimeria pogonae comb. nov. (Apicomplexa: Eimeriidae).

    Science.gov (United States)

    Szczepaniak, Klaudiusz Oktawian; Tomczuk, Krzysztof; Lojszczyk-Szczepaniak, Anna; Lopuszynski, Wojciech

    2016-02-01

    The presented paper provides a reclassification of Eimeria pogonae from Pogona vitticeps into the correct genus Choleoeimeria. A description of exogenous and endogenous stages of biliary coccidium is given. Sporulation of the oocysts was endogenous. The mature oocysts contained four sporocysts each with two sporozoites. Oocysts were ellipsoidal in shape, with average length/width ratio 1.7 and measured 28.4 (SD1.5) × 16.8 (SD 1.5). The micropyle, residuum, and polar granules were absent from the sporulated oocysts. Ovoidal in shape, sporosysts without Steida bodies contained residuum and two elongated and boat-shaped sporozoites. The endogenous stages of the coccidia were located mainly in the epithelium of bile ducts; however, single-epithelium cells of the gallbladder were also infected.

  16. Comparison of toltrazuril and sulphadimethoxine in the treatment of intestinal coccidiosis in pet rabbits.

    Science.gov (United States)

    Redrobe, S P; Gakos, G; Elliot, S C; Saunders, R; Martin, S; Morgan, E R

    2010-08-21

    Treatment of intestinal coccidiosis was studied in domestic pet rabbits. In 45 rabbits aged four months or more, coccidial oocysts were observed in the faeces of 35 rabbits at a mean density of 806 opg (range 50 to 6800 opg). Eimeria magna was the dominant species, with Eimeria media and Eimeria intestinalis also being common. The presence of the hepatic species Eimeria stiedae was not recorded. A single oral dose of 2.5 mg/kg or 5.0 mg/kg toltrazuril, or a single oral dose of 50 mg/kg sulphadimethoxine followed by its inclusion in drinking water at 1 g/4 l for nine days, were all found to significantly reduce the faecal oocyst count by 73 to 99 per cent. The extent of oocyst reduction in the faeces was not dependent on the dose of toltrazuril. Oocyst counts began to rise again in the days after treatment ceased.

  17. Caryospora uptoni n. sp. (Apicomplexa: Eimeriidae) from red-tailed hawks (Buteo jamaicensis borealis).

    Science.gov (United States)

    Lindsay, D S; Blagburn, B L

    1986-10-01

    Oocysts of Caryospora uptoni n. sp. were described from the feces of red-tailed hawks, Buteo jamaicensis borealis. Sporulated oocysts were spherical or subspherical and measured 28.1 by 26.4 micron. The oocyst wall was composed of a yellowish outer layer and brownish inner layer and was about 1.5 micron thick. Neither micropyle, polar granules, nor oocyst residuum were present. A single, spherical sporocyst 18.2 by 17.9 micron was present; a Stieda body was absent. A spherical eccentrically located sporocyst residuum was present in many sporocysts, but it degenerated to form a dispersed granular residuum in other sporocysts. Eight randomly arranged sporozoites, 12.6 by 4.2 micron, were present in each sporocyst; they contained a centrally or slightly posteriorly located nucleus.

  18. Caryospora uptoni and Frenkelia sp.-like coccidial infections in red-tailed hawks (Buteo borealis).

    Science.gov (United States)

    Lindsay, D S; Blagburn, B L

    1989-07-01

    The feces from 16 red-tailed hawks (Buteo borealis) were examined by fecal flotation for the presence of coccidial oocysts or sporocysts. Oocysts of Caryospora uptoni were found in five (31%), sporocysts of a Frenkelia sp.-like coccidium were found in eight (50%), and mixed infections with both species of coccidia were found in three (19%) red-tailed hawks. Neither oocysts nor sporocysts were found in six (38%) red-tailed hawks. Sexual stages of C. uptoni were found in the duodenum and jejunum of a naturally infected red-tailed hawk. Sexual stages were located in enterocytes on the distal three-fourths of the villi. This study shows that over 60% of red-tailed hawks may be passing coccidial oocysts/sporocysts in their feces and provides morphological information for diagnosing C. uptoni infections in histological sections.

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

    Science.gov (United States)

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

    2010-03-01

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

  20. 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...

  1. 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...

  2. Experimental toxoplasmosis in pregnant bitches.

    Science.gov (United States)

    Bresciani, K D; Costa, A J; Toniollo, G H; Sabatini, G A; Moraes, F R; Paulillo, A C; Ferraudo, A S

    1999-09-30

    Three Toxoplasma gondii free bitches (No. 3-5) were fed 15000 sporulated T. gondii oocysts at 56, 40 and 32 day of gestation and the outcome of the pregnancy was monitored. Two of the three dogs infected during pregnancy showed evidence of congenital infection and one aborted. Two control bitches not fed oocysts delivered eight uninfected healthy pups. This study demonstrated that T. gondii can be congenitally transmitted in dogs when bitches are infected during pregnancy.

  3. Diversity of Eimeria spp. in dairy cattle of Guwahati, Assam, India

    OpenAIRE

    Das, M.; Deka, D. K.; Sarmah, P. C.; Islam, S.; S Sarma

    2015-01-01

    Aim: To determine the prevalence and diversity of Eimeria spp. in dairy cattle present in and around Guwahati, Kamrup district, Assam, India. Materials and Methods: A total of 2339 fecal samples of calves (535), heifer (641) and adult (1163) cattle were screened for 1 year present in and around Guwahati, Assam for detection of Eimeria oocysts by flotation techniques. Sporulation of the oocyst was done in 2.5% potassium dichromate solution for identification of the Eimeria species. Results: Ex...

  4. THE INHIBITORY EFFECT OF ACRIDINE ON THE SPOROGONY OF A COCCIDIUM (EIMERIA STIEDAE).

    Science.gov (United States)

    Lewis, P A

    1924-07-31

    The development or ripening of the oocyst of the coccidium of the rabbit is prevented by acridine hydrochloride provided that the cysts are exposed to the action of the chemical before development has started. After sporoblasts are formed acridine does not prevent further development. Many other substances, some of them known to be active against certain protozoan parasites, have no influence on the ripening of the oocysts of the coccidium.

  5. Bioaccumulation of Toxoplasma and Cryptosporidium by the freshwater crustacean Gammarus fossarum: Involvement in biomonitoring surveys and trophic transfer.

    Science.gov (United States)

    Bigot-Clivot, Aurélie; Palos Ladeiro, Mélissa; Lepoutre, Alexandra; Bastien, Fanny; Bonnard, Isabelle; Dubey, Jitender P; Villena, Isabelle; Aubert, Dominique; Geffard, Olivier; François, Adeline; Geffard, Alain

    2016-11-01

    The protozoa Toxoplasma gondii and Cryptosporidium parvum are public health priorities because their oocysts can persist in recreational, surface, drinking, river, and sea water sources for a long time. To evaluate the capacity of the freshwater crustacean Gammarus fossarum to accumulate T. gondii and C. parvum oocysts, gammarids were exposed to 200, 2000 or 20,000 oocysts per gammarid and per day for 21 days followed by 5 days of depuration. C. parvum DNA was detected by qPCR in G. fossarum in only one out of four pools for the highest concentration and after 14 days of exposure, and T. gondii DNA was detected after 7 days of exposure to the two highest concentrations. Our results document the capacity of G. fossarum to accumulate T. gondii in its tissues proportionally to the ambient concentration; the maximum number of oocysts was detected in gammarid tissues after exposure to 20,000 oocysts per day. Mean values of 3.26 (±3), 21.71 (±15.18), and 17.41 (±10.89) oocysts were detected in gammarids after 7, 14, and 21 days, respectively, and after 5 days of depuration, T. gondii oocysts were still present in gammarid tissues. These results show for the first time that a freshwater crustacean can bioaccumulate T. gondii oocysts, suggesting that G. fossarum is a potential effective bioindicator of protozoan contamination in biomonitoring studies. Moreover, due to its key position in freshwater food webs, G. fossarum could also play a role in the trophic transfer of protozoa. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Eimeripain, a Cathepsin B-Like Cysteine Protease, Expressed throughout Sporulation of the Apicomplexan Parasite Eimeria tenella

    OpenAIRE

    Anaïs Rieux; Simon Gras; Fabien Lecaille; Alisson Niepceron; Marilyn Katrib; Smith, Nicholas C.; Gilles Lalmanach; Fabien Brossier

    2012-01-01

    The invasion and replication of Eimeria tenella in the chicken intestine is responsible for avian coccidiosis, a disease that has major economic impacts on poultry industries worldwide. E. tenella is transmitted to naïve animals via shed unsporulated oocysts that need contact with air and humidity to form the infectious sporulated oocysts, which contain the first invasive form of the parasite, the sporozoite. Cysteine proteases (CPs) are major virulence factors expressed by protozoa. In this ...

  7. Eimeria fraterculae sp. n. in the kidneys of Atlantic puffins (Fratercula arctica) from Newfoundland, Canada: species description and lesions.

    Science.gov (United States)

    Leighton, F A; Gajadhar, A A

    1986-10-01

    Renal coccidiosis is reported for the first time in an auk (Alcidae). Infection was detected in seven of 50 nestling Atlantic puffins (Fratercula arctica) and a new species of coccidia, Eimeria fraterculae sp. n., is described. The structure and sporulation of oocysts are characterized. Meronts, gamonts, and developing oocysts were present in collecting duct epithelium of medullary cones. The predominant host response was hypertrophy of infected cells, tubule dilation, and a mild localized peritubular infiltration with mononuclear inflammatory cells.

  8. [Construction of subtractive cDNA libraries of the sporogony stage of Eimeria tenella by suppression subtractive hybridization].

    Science.gov (United States)

    Han, Hong-Yu; Lin, Jiao-Jiao; Zhao, Qi-Ping; Dong, Hui; Jiang, Lian-Lian; Wang, Xin; Han, Jing-Fang; Huang, Bing

    2007-11-01

    In order to clone and identify differentially expressed genes in the sporogony stage of Eimeria tenella, the cDNAs from unsporulated oocysts and sporulated oocysts of E. tenella were used as driver, respectively, the cDNAs from sporozoites of E. tenella was used tester, Two subtractive cDNA libraries of sporozoites were constructed by using the technique of suppression subtractive hybridization (SSH). the cDNAs from unsporulated oocysts was used driver, the cDNAs from sporulated ooceysts was used tester, one subtractive cDNA library of sporulated oocysts was constructed. PCR amplification revealed that the two subtractive cDNA libraries of sporozoites and one subtractive cDNA library of sporulated oocysts contained approximated 96%, 96% and 98% recombinant clones, respectively. Fifty positive clones were sequenced and analyzed in GenBank with Blast search from three subtractive cDNA libraries, respectively, thirteen unique sequences were found from the subtractive cDNA library of sporulated oocysts, eight ESTs shared significant identity with previously described. A total of forty unique sequences were obtained from the two subtractive cDNA libraries, nine ESTs shared significant identity with previously described, the other sequences represent novel genes of E. tenella with no significant homology to the proteins in Genbank. These results have provided the foundation for cloning new genes of E. tenella and further studying new approaches to control coccidiosis.

  9. Eimeria galateai sp. n. from the Paradise Kingfisher, and Eimeria duncani sp. n. from the Sacred Kingfisher in Papua New Guinea.

    Science.gov (United States)

    Varghese, T

    1977-05-01

    Eimeria galateai sp. n. from the paradise kingfisher (Tanysiptera galatea Gray) and Eimeria duncani sp. n. from the sacred kingfisher (Halcyon sancta Vigors & Horsfield) have been described from Papua New Guinea. Four of 11 paradise kingfishers were infected with E. galateai oocysts, measuring 13 (11-16) X 9 (8-11) micron. The oocysts were ovoid with nipple-like protrusion at one pole. Micropyle and polar granule were absent, while oocyst residuum (5 X 4 micron) was present. Sporocysts, measuring 5 (4-6) X 2 micron, were elongate-ovoid, and had a distinct convex Stieda body; the sporocyst residuum was absent. Two of 9 sacred kingfishers were infected with ovoid-truncated, 22 (19-25) X 16 (12-18) micron oocysts of E. duncani. Polar granule (5 X 2) was present in the oocysts, but there was no micropyle or oocyst residuum. Sporocysts were ovoid, measuring 9 (8-10) X 5 (4-6) micron, with a prominent Stieda body, and granular sporocyst residuum. Eimeria galateai and E. duncani are the first species of this genus to be described from birds of the order Coraciiformes.

  10. Immunization of young chicks using graded dose of wild strain of Eimeria tenella

    Directory of Open Access Journals (Sweden)

    E.N. Kimbita

    2005-09-01

    Full Text Available A wild strain of Eimeria tenella was isolated and utilized for immunization studies. Its optimal sporulation was attained at room temperature 24-25 °C after 24-48 h. Two groups of chicks were immunized by dosing a graded dose of five oocysts/chick/day for 6 days followed by 50 oocysts/chick/day for 7 days. A third group was not immunized and served as a negative control. Immunized chicks gained mass at the same rate as unimmunized ones, but when challenged with 200 000 oocysts/chick, mass gains declined in the unimmunized group. The growth rate of immunized chicks was not affected by challenge (P > 0.05. Upon challenge, unimmunized chicks produced significantly more oocysts than immunized chicks (P < 0.005. Immunized chicks withstood a challenged with 200 000 oocysts/chick without developing any clinical signs whereas the unimmunized chicks developed typical clinical signs of coccidiosis. Unimmunized chicks had significantly more severe lesions in the caecum than any other group (P > 0.005 and also produced significantly more oocysts than any other group (P > 0.005.

  11. Protective effect of Eimeria stiedae coproantigen against hepatic coccidiosis in rabbits.

    Science.gov (United States)

    Abdel Megeed, Kadria N; Abuel Ezz, Nadia M T; Abdel-Rahman, Eman H

    2005-08-01

    Immunization of rabbits against hepatic coccidiosis was tried. The animals were immunized twice with Eimeria stiedae coproantigen in freund's adjuvant with two week intervals. The rabbits were challenged orally with sporulated E. stiedae oocysts one week post last injection. The protection was assessed by number of oocysts output, number of focal liver lesions, clinical sings and antibody response. The immunization resulted in 70% protection from infection and decline in oocysts count. High level of IgG response in immunized rabbits than control infected ones was occurred and being responsible for the recorded protection. The electrophoretic make up of the coproantigen and oocyst antigen showed different patterns of separation. Common as well as specific bands to each antigen were identified. Using the rabbit sera after 3 weeks post vaccination in immunoblot assay, immunogenic components were detected of molecular weight 155, 103, 74, 66, 44, 22KD with coproantigen and 155, 115, 57, 26 KD with oocyst antigen. While, the rabbit sera after 2 and 4 weeks post challenge reacted with oocyts antigen, in immunoblot assay, revealing immunogenic bands of molecular weight 155, 115, 57, 26, 22KD and 155, 115, 57, 25 KD respectively. Bands of 22KD and 155KD are partially responsible for eliciting host protective immune response where they were recognized by immunized sera in coproantigen and by immunized infected sera in oocyst antigen.

  12. Chitosan and metal salt coagulant impacts on Cryptosporidium and microsphere removal by filtration.

    Science.gov (United States)

    Brown, Trevor J; Emelko, Monica B

    2009-02-01

    Maintenance of appropriate chemical pretreatment is a critical component of ensuring proper filtration performance. Pilot-scale in-line filtration studies were performed to investigate the relative impacts of chitosan, alum, and FeCl(3) coagulation on the removal of Cryptosporidium parvum oocysts and oocyst-sized polystyrene microspheres by granular media filtration. Similar removals of oocysts and microspheres were achieved when optimal coagulant doses were utilized. Sub-optimal alum and FeCl(3) coagulation resulted in a deterioration filter effluent turbidity (0.2-0.3NTU) and total particle counts (30-100 total particles > or =2microm/mL) that were accompanied by reduced (by approximately 2-3-log) median oocyst and microsphere removals by filtration. At all doses investigated, chitosan coagulation resulted in excellent turbidity and particle reductions by filtration. Nonetheless, chitosan coagulation at doses of 0.1, 0.5, and 1.0mg/L did not result in appreciable improvements in C. parvum oocyst removal relative to complete coagulation failure (median oocyst removals were alum and FeCl(3) coagulation, but not chitosan coagulation.

  13. Observations on some Avian Coccidia (Apicomplexa: Eimeriidae in Amazonian Brazil

    Directory of Open Access Journals (Sweden)

    Ralph Lainson

    1994-09-01

    Full Text Available Oocysts of Eimeria porphyrulae n. sp. are described in faeces of Porphyrula martinica (Aves: Gruiformes: Rallidae. They are ellipsoidal to oval, 22.4 x 17.7 (20.0-23.7 x 16.2-18.7 µm, shape-index (length/width 1.3. Oocyst wall about 1.25 µm thick, colourless, with two layers: inner one prominently striated. Micropyle and sub-micropylar granule present: no oocyst residuum. Sporocysts 17.5 x 9.0 (17.0-19.0 x 8.0-10.0 µm, shape-index 1.9, with inconspicuous Stieda/sub-Stieda bodies. Sporocyst residuum of scattered granules, sometimes a compact mass: sporozoites with two refractile bodies. Eimeria crypturelli n. sp. is described in faeces of Crypturellus soiu (Tinamiformes: Tinamidae. Oocysts ellipsoidal-oval, 20.75 x 14.5 (17.5-25.0 x 11.25-21.25 µm, shape-index 1.4. Oocysts wall about 1.25 µm thick and bi-layered: inner layer faintly striated. Micropyle present, with oocyst residuum immediately below: single polar body rarely present. Sporocysts 13.0 x 7.5 (12.5-13.75 x 7,5-8.1 µm, shape-index 1.7, with a Stieda body but seemingly no sub-Stieda. Sporocyst residuum compact: sporozoites with two refractile bodies. Isospora cacici n. sp. is recorded from faeces of Cacicus cela cela (Passeriformes: Icteridae. Oocysts subspherical-spherical, 26.5 x 23.7 (22.5-27.5 x 20.0-26.2 µm, shape-index 1.1. Wall a single, colourless layer about 1.5 µm thick. No micropyle or oocyst residuum: 1-2 polar bodies. Sporocysts ellipsoidal, 17.7 x 12.5 (17.5-18.75 x 11.25-13.75 µm, shape-index 1.4, with pronounced Stieda/sub-Stieda bodies: residuum compact and sporozoites with two refractile bodies. Isospora thraupis n. sp. is described from faeces of Thraupis palmarum melanoptera (Passeriformes: Thraupidae. Oocysts subspherial-spherical, 19.9 x 19.0 (18.7-21.2 x 18.75-20.0 µm, shape-index 1.0. Wall about 0.6 µm thick, smooth, colourless and a single layer: no micropyle, oocyst residuum or polar bodies. Sporocysts 14.2 x 9.2 (13.7-16.2 x 8.7-10.0 µm, shape

  14. 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

    Directory of Open Access Journals (Sweden)

    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

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

    Science.gov (United States)

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

    2010-01-01

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

  16. Shedding of live Eimeria vaccine progeny is delayed in chicks with delayed access to feed after vaccination.

    Science.gov (United States)

    Price, Kayla R; Freeman, Megan; Van-Heerden, Kobus; Barta, John R

    2015-03-15

    Hatching, processing and transportation result in inevitable delays before chicks are placed into brooding and receive their first feed and drinking water after hatching. To determine if delayed access to feed for different durations following live Eimeria vaccination affected initial shedding of vaccine progeny, replacement layer chicks (480, Lohmann-LSL Lite) aged approximately 6h after hatch were administered a commercial live Eimeria vaccine. Vaccinated chicks were divided randomly into groups and were provided access to feed immediately (0 h) or after a delay of 6, 12, or 24 h (4 treatments × 6 replicates per treatment × 20 pullets per replicate). All pullets were provided drinking water immediately following vaccination. Fecal oocysts shed per gram of feces for each cage replicate was determined daily from 4 to 9 days post inoculation. Chicks provided feed immediately had peak oocyst shedding at 5 days post-inoculation but delayed access to feed for 24h was associated with a 2 days delay in peak oocyst shedding to 7 days post-inoculation. Chicks with delays in access to feed of intermediate duration (i.e. 6 or 12h) had peak oocyst shedding at 6 days post-inoculation. Overall oocyst shedding was not affected. Live Eimeria vaccination success may be measured by evaluating initial shedding of oocysts at some pre-established time after vaccine application, usually by a single fecal collection conducted at 5, 6 or 7 days post-inoculation. Recognizing that withholding feed following live Eimeria vaccination shifts the time of the resultant peak oocyst shedding complicates the assessment of vaccine application; if delayed access to feed is not taken into account, it is possible that false conclusions could be drawn regarding the relative success of vaccine administration.

  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. Experimentally induced clinical Cystoisospora canis coccidiosis in dogs with prior natural patent Cystoisospora ohioensis-like or C. canis infections.

    Science.gov (United States)

    Houk, Alice E; O'Connor, Thomas; Pena, Hilda F J; Gennari, Solange Maria; Zajac, Anne M; Lindsay, David S

    2013-10-01

    Diarrhea caused by intestinal coccidia (Cystoisospora species) is a common problem in pet dogs and in dogs in animal shelters. Cystoisospora canis has the largest oocysts of the 4 named species of coccidia infecting dogs. The present study examined an isolate of C. canis obtained from a dog from São Paulo, SP, Brazil. Oocysts sporulated within 2 days at room temperature, and 20 sporulated oocysts were measured at 37.6 by 28.6 μm (range 35-42 by 26-31 μm). Most sporulated oocysts contained 2 sporocysts, each with 4 sporozoites, although a few (coccidiosis using sulfadimethoxine was recommended. Dog CRU had a natural C. canis infection and did not develop clinical disease after oral infection with C. canis oocysts. This dog had a prepatent period of 9 days and a patent period of 3 days, corresponding to experimental infection with the new isolate of C. canis. It excreted fewer C. canis oocysts than did the other dogs. The 4 dogs with natural C. ohioensis-like infection all developed clinical disease, and 1 required treatment. The prepatent period was 9-10 days, and the patent period was 10-11 days in these dogs. All 6 dogs not naturally infected with Cystoisospora developed clinical disease, and 2 required treatment. The prepatent period was 9-10 days, and the patent period was 8-12 days. The present study confirms that C. canis is a primary pathogen for young dogs. It demonstrates that prior infection with C. canis but not C. ohioensis-like coccidia confers some resistance to clinical disases and a decrease in oocyst production in dogs challenged with C. canis.

  19. Activity of diclazuril against coccidiosis in growing rabbits: experimental and field experiences

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    Monita Vereecken

    2012-12-01

    Full Text Available The efficacy of diclazuril in growing rabbits was investigated under experimental and field conditions. In a first experimental trial, the susceptibility of recent isolated French Eimeria field strains to in-feed use of diclazuril, salinomycin and robenidine was studied in fattening rabbits. Rabbits were challenged at the age of 31 d with a mixed inoculum of Eimeria magna, E. media and E. perforans. Production data and oocyst excretion were compared with an infected-untreated control group and an uninfected-untreated control group. Infection resulted in significantly lower production data and higher oocyst excretion in the infected-untreated control group. Salinomycin and diclazuril treated rabbits were able to control the infection, demonstrated also by comparable weight gain and final weight to those of the uninfected-untreated control rabbits and significantly higher than those of the infected-untreated control rabbits. Based on the production data and oocyst excretion, robenidine was not able to control the infection adequately. Economic performance (weight gain, feed intake, feed conversion and oocyst excretion were significantly worse than in the uninfected-untreated controls. In a second trial, a 1 yr longitudinal study was carried out in Italy to evaluate the excretion of coccidia in growing rabbits from 8 meat farms applying a 2-phase anticoccidial programme (diclazuril and robenidine. Parasitological parameters (oocyst counts and species identification were measured monthly. Seven of the 11 known coccidial rabbit species were identified. Variable levels of oocysts per gram were detected in the farms, but on all farms lower oocyst per gram and a reduced number of Eimeria spp. in rabbit faeces were recorded in the 8-mo treatment period with diclazuril.

  20. Coccidia of the collared peccary (Tayassu tajacu) in southern Texas with descriptions of three new species of Eimeria (Apicomplexa: Eimeriidae).

    Science.gov (United States)

    Wilber, P G; Hellgren, E C; Gabor, T M

    1996-08-01

    In February 1993, fresh fecal samples were collected from 47 collared peccaries (Tayassu tajacu) killed by hunters at the Chaparral Wildlife Management Area, southern Texas, USA. Five species of coccidia (Eimeria chaparralensis n. sp. [9/47, 19%], Eimeria dicotylensis n. sp. [21/47, 21%], Eimeria pecari n. sp. [2/47, 4%], Eimeria sp. [1/47, 2%], and Klossia sp. [1/47, 2%]) were observed. Measurements are in micron. Sporulated oocysts of E. chaparralensis are rough-walled, elongate ovoidal, 43.3 x 28.5 (37-52 x 26-35); sporocysts are elongate ellipsoidal 21.8 x 9.0 (16-27 x 7-12); micropyle (approximately 4.9), Stieda, and substieda body are present; sporocyst residuum is present in newly sporulated oocysts; polar granule and oocyst residuum are absent. Sporulated oocysts of E. dicotylensis are smooth-walled, ovoidal, 25.7 x 20.1 (23-29 x 17-23); sporocysts are ellipsoidal 13.0 x 6.9 (11-17 x 6-9); micropyle and oocyst residuum are absent; polar body sometimes present; Stieda body and sporocyst residuum always present. Sporulated oocysts of E. pecari are smooth-walled, elongate ellipsoidal, 26.8 x 18.1 (22-31 x 15-21); sporocysts are elongate ellipsoidal 16.4 x 5.9 (13-22 x 4-7); micropyle (approximately 5.8) with collar, Stieda body, substieda body, and sporocyst residuum are present; polar granule and small oocyst residuum sometimes present. There were no sex or age differences in prevalences of infection, and there were no positive or negative associations between any species of eimerian. The majority of hosts were infected with a single species of Eimeria. Overall prevalence of infection with eimerians was 23/47 (49%).

  1. An experimental Toxoplasma gondii dose response challenge model to study therapeutic or vaccine efficacy in cats.

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    Jan B W J Cornelissen

    Full Text Available High numbers of Toxoplasma gondii oocysts in the environment are a risk factor to humans. The environmental contamination might be reduced by vaccinating the definitive host, cats. An experimental challenge model is necessary to quantitatively assess the efficacy of a vaccine or drug treatment. Previous studies have indicated that bradyzoites are highly infectious for cats. To infect cats, tissue cysts were isolated from the brains of mice infected with oocysts of T. gondii M4 strain, and bradyzoites were released by pepsin digestion. Free bradyzoites were counted and graded doses (1000, 100, 50, 10, and 250 intact tissue cysts were inoculated orally into three cats each. Oocysts shed by these five groups of cats were collected from faeces by flotation techniques, counted microscopically and estimated by real time PCR. Additionally, the number of T. gondii in heart, tongue and brains were estimated, and serology for anti T. gondii antibodies was performed. A Beta-Poisson dose-response model was used to estimate the infectivity of single bradyzoites and linear regression was used to determine the relation between inoculated dose and numbers of oocyst shed. We found that real time PCR was more sensitive than microscopic detection of oocysts, and oocysts were detected by PCR in faeces of cats fed 10 bradyzoites but by microscopic examination. Real time PCR may only detect fragments of T. gondii DNA without the presence of oocysts in low doses. Prevalence of tissue cysts of T. gondii in tongue, heart and brains, and anti T. gondii antibody concentrations were all found to depend on the inoculated bradyzoite dose. The combination of the experimental challenge model and the dose response analysis provides a suitable reference for quantifying the potential reduction in human health risk due to a treatment of domestic cats by vaccination or by therapeutic drug application.

  2. An Experimental Toxoplasma gondii Dose Response Challenge Model to Study Therapeutic or Vaccine Efficacy in Cats

    Science.gov (United States)

    Cornelissen, Jan B. W. J.; van der Giessen, Joke W. B.; Takumi, Katsuhisa; Teunis, Peter F. M.; Wisselink, Henk J.

    2014-01-01

    High numbers of Toxoplasma gondii oocysts in the environment are a risk factor to humans. The environmental contamination might be reduced by vaccinating the definitive host, cats. An experimental challenge model is necessary to quantitatively assess the efficacy of a vaccine or drug treatment. Previous studies have indicated that bradyzoites are highly infectious for cats. To infect cats, tissue cysts were isolated from the brains of mice infected with oocysts of T. gondii M4 strain, and bradyzoites were released by pepsin digestion. Free bradyzoites were counted and graded doses (1000, 100, 50, 10), and 250 intact tissue cysts were inoculated orally into three cats each. Oocysts shed by these five groups of cats were collected from faeces by flotation techniques, counted microscopically and estimated by real time PCR. Additionally, the number of T. gondii in heart, tongue and brains were estimated, and serology for anti T. gondii antibodies was performed. A Beta-Poisson dose-response model was used to estimate the infectivity of single bradyzoites and linear regression was used to determine the relation between inoculated dose and numbers of oocyst shed. We found that real time PCR was more sensitive than microscopic detection of oocysts, and oocysts were detected by PCR in faeces of cats fed 10 bradyzoites but by microscopic examination. Real time PCR may only detect fragments of T. gondii DNA without the presence of oocysts in low doses. Prevalence of tissue cysts of T. gondii in tongue, heart and brains, and anti T. gondii antibody concentrations were all found to depend on the inoculated bradyzoite dose. The combination of the experimental challenge model and the dose response analysis provides a suitable reference for quantifying the potential reduction in human health risk due to a treatment of domestic cats by vaccination or by therapeutic drug application. PMID:25184619

  3. Morphological and molecular characterization of Eimeria paludosa coccidian parasite (Apicomplexa:Eimeriidae) in a dusky moorhen (Gallinula tenebrosa, Gould, 1846) in Australia.

    Science.gov (United States)

    Yang, Rongchang; Brice, Belinda; Elloit, Aileen; Lee, Elvina; Ryan, Una

    2014-12-01

    An Eimeria species is described from a dusky moorhen (Gallinula tenebrosa). Sporulated oocysts (n = 40) are ovoid, with a pitted single-layered oocyst wall in young oocysts and a relatively smooth wall in the mature oocysts. Oocyst wall was 1.0 µm thick, oocysts measured 17.3 × 13.3 (16.3-17.9 × 12.7-13.9) µm, oocyst length/width (L/W) ratio, 1.3. Oocyst residuum was absent. A large polar granule was always observed in the centre of the micropyle and many small polar granules were observed when the focus was on the wall. Sporocysts are elongate-ovoid, 8.4 × 5.1 (8.0-8.9 × 4.9-5.5) µm, sporocyst L/W ratio, 1.6 (1.5-1.8), sporocyst residuum was present, composed of numerous granules in a spherical or ovoid mass. Each sporocyst contained 2 elongate sporozoites, 7.7 × 2.6 (7-10 × 2.2-3) µm. A spherical-ellipsoid posterior refractile body was found in the sporozoites. A nucleus is located immediately anterior to the posterior refractile body. When the oocyst measurements and features were compared with valid Eimeria species from hosts in the Rallidae family, this Eimeria species was identified as E. paludosa. This is the first report of E. paludosa in Australia and the dusky moorhen (Gallinula tenebrosa) in a new host for this species. Molecular analysis was conducted at three loci; the 18S and 28S ribosomal RNA genes and the mitochondrial cytochrome oxidase gene (COI). At the 18 S locus, E. paludosa shared 97.3% genetic similarity with Eimeria gruis (GenBank accession number: AB544336). It also shared 99.2% genetic similarity with Eimeria crecis (GenBank accession numbers: HE653904 and HE653905) and 98.5% similarity with Eimeria nenei (GenBank accession numbers: HE653906), both of which were identified from a corncrake (Crex crex) in the United Kingdom. At the 28S locus, E. paludosa shared 91.4% similarity with E. papillata from a chicken (Gallus gallus) in the USA. At COI locus, E. paludosa was in a clade by itself and shared 87.2% similarity with E

  4. Comparison of different diagnostic techniques for the detection of cryptosporidiosis in bovines

    Directory of Open Access Journals (Sweden)

    H. K. M. Rekha

    2016-02-01

    Full Text Available Aim: Aim of the present study was to compare different methods, viz., Sheather’s sugar flotation (SSF, Ziehl-Neelsen (ZN, Kinyoun’s acid-fast method (KAF, safranin-methylene blue staining (SMB, and negative staining techniques such as nigrosin staining, light green staining, and malachite green staining for the detection of Cryptosporidium spp. oocysts in bovines. Materials and Methods: A total of 455 fecal samples from bovines were collected from private, government farms and from the clinical cases presented to Department of Medicine, Veterinary College, Bengaluru. They were subjected for SSF, ZN, KAF, SMB and negative staining methods. Results: Out of 455 animal fecal samples screened 5.71% were found positive for Cryptosporidium spp. oocysts. The species were identified as Cryptosporidium parvum in calves and Cryptosporidium andersoni in adults based on the morphological characterization and micrometry of the oocysts. Conclusions: Of all the techniques, fecal flotation with sheather’s was found to be more specific and sensitive method for the detection of Cryptosporidium spp. oocysts. Among the conventional staining methods, the SMB gives better differentiation between oocysts and yeast. Among the three negative staining methods, malachite green was found sensitive over the other methods.

  5. Deformability Assessment of Waterborne Protozoa Using a Microfluidic-Enabled Force Microscopy Probe.

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    John S McGrath

    Full Text Available 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 Cryptosporidium in drinking-water do not discern between non-pathogenic and pathogenic species, nor between viable and non-viable oocysts. Using FluidFM, a novel force spectroscopy method employing microchannelled cantilevers for single-cell level manipulation, we assessed the size and deformability properties of two species of Cryptosporidium that pose varying levels of risk to human health. A comparison of such characteristics demonstrated the ability of FluidFM to discern between Cryptosporidium muris and Cryptosporidium parvum with 86% efficiency, whilst using a measurement throughput which exceeded 50 discrete oocysts per hour. In addition, we measured the deformability properties for untreated and temperature-inactivated oocysts of the highly infective, human pathogenic C. parvum to assess whether deformability may be a marker of viability. Our results indicate that untreated and temperature-inactivated C. parvum oocysts had overlapping but significantly different deformability distributions.

  6. Human cryptosporidiosis: detection of specific antibodies in the serum by an indirect immunofluorescence

    Directory of Open Access Journals (Sweden)

    Braz Lúcia M.A.

    1996-01-01

    Full Text Available Cryptosporidium sp., a coccidian parasite usually found in the faeces of cattle, has been recently implicated as an agent of human intestinal disease, mainly in immunocompromised patients. In the study realized, by an indirect immunofluorescence technique, specific immunoglobulins (IgG and IgM have been demonstrated in human serum against Cryptosporidium oocysts. Purified oocysts were used as antigens in the indirect immunofluorecence assay. After analyzing this test in sera from selected groups of patients, the frequency of both specific IgG and IgM of immunocompetent children who were excreting oocysts in their faeces was 62% and in children with negative excretion of oocysts was 20% and 40%, respectively. In adults infected with the human immunodeficiency virus (HIV and who were excreting Cryptosporidium in their stools, the frequency was 57% for IgG but only 2% for IgM. Twenty three percent of immunocompromised adults with not determined excretion of oocysts in their stools had anti-Cryptosporidium IgG in their sera. Children infected with human immunodeficiency virus had no IgM and only 14% had IgG detectable in their sera. The indirect immunoflorescence assay, when used with other parasitological techniques appears to be useful for retrospective population studies and for diagnosis of acute infection. The humoral immune response of HIV positive patients to this protozoan agent needs clarification.

  7. Chlamydotis macqueenii and C. undulata (Aves: Otididae) are new hosts for Caryospora megafalconis (Apicomplexa: Eimeriidae) and proposal of the genus Avispora gen. nov.

    Science.gov (United States)

    Schuster, Rolf K; Woo, Patrick C Y; Poon, Rosana W S; Lau, Susanna K P; Sivakumar, Saritha; Kinne, Jörg

    2016-11-01

    Oocysts of a coccidian morphologically matching features of Caryospora megafalconis Klüh, 1994 were found in fecal samples and contents of the large intestines in five wild caught Clamydotis macqueenii (Gray) and 19 captive bred C. undulata (Jaquin). Scrapings of the intestinal mucosa of necropsied birds revealed macrogamonts and unsporulated oocysts. Sporulation in a potassium dichromate solution at 26 °C was completed in 48 h. Intestinal contents and sporulated oocysts obtained from feces of infected bustards as well as sporulated oocysts of C. megafalconis and C. neofalconis Böer, 1982 from two Falco rusticolis Linnaeus and one F. peregrinus Tunstall were used for DNA sequencing of the cox1, 18S ribosomal ribonucleic acid (rRNA), and 28S rRNA genes. The phylogenetic trees for all three genes showed that sequences of the material from bustards were identical with C. megafalconis from falcons. C. neofalconis and C. daceloe Yang et al., 2014 were situated in the neighboring clades. Contrary to this, subsequent sequences of C. bigenetica Wacha and Christiansen, 1982 from rattlesakes are at a distinct distance suggesting that despite morphological similarities of the oocysts, there are differences between Caryospora species of birds and reptiles. For this reason, it might be reasonable to transfer avian Caryospora species into a new genus Avispora.

  8. A new Eimeria species (Protozoa: Eimeriidae) from caribou in Ameralik, West Greenland.

    Science.gov (United States)

    Skirnisson, K; Cuyler, C

    2016-04-01

    Fecal samples of 11 calves shot in the Ameralik area, West Greenland, in August-September 2014 were examined for coccidian parasites. The calves belonged to a population of interbreeding indigenous caribou Rangifer tarandus groenlandicus and feral semi-domestic Norwegian reindeer Rangifer tarandus tarandus. Two coccidian species were found: Eimeria rangiferis and a coccidium that was identified and described as a new species. The latter's sporulated oocyst is spherical or slightly subspherical. Average size is 25.6 × 24.8 μm. The oocyst has two distinct walls. Wall thickness is ∼1.4 μm. The unicolored outer wall is brown, the inner wall is dark gray. The oocysts contain a small polar granule but are devoid of a microphyle. The oocysts enclose four ovoid-shaped sporocysts with a rounded end opposite to the Stieda body. The average size of sporocysts is 15.2 × 7.8 μm. Sporocysts contain a granular sporocyst residuum that forms a spherical cluster between the sporocysts, one large refractile body is present in each sporozoite. The spherical form easily distinguishes oocysts of the new species from the seven previously described eimerid species in R. tarandus. This is the first eimerid described as a new species to the sciences from caribou in the Nearctic.

  9. Observation of the excretion pattern of a precocious line of Eimeria necatrix and the strengthening of immune homogeneity.

    Science.gov (United States)

    Chen, Rui-Ai; Wang, Wen-Li; Lin, Rui-Qing; Liu, Li-Dan; Liao, Gui-Cheng; Tang, Jian-Hong; Wu, Song-Ming; Tan, Zhi-Jian; Weng, Ya-Biao

    2016-05-01

    The excretion frequencies of cecal and intestinal droppings of Chinese Lingnan yellow chickens were observed for 10 consecutive days. The chickens were then orally inoculated with a precocious line of Eimeria necatrix, and the oocysts present in the cecal and intestinal droppings were separately collected and monitored using the McMaster method. The results showed that the excretion frequency of cecal droppings was significantly lower than that of intestinal droppings, and the oocysts of E. necatrix were distributed primarily in the cecal droppings. This distribution affects the homogeneity of the second and third generation of oocysts ingested by the chickens and therefore affects the immune effect observed during E. necatrix immunization. To artificially strengthen the immunologic homogeneity against E. necatrix, a method of artificially strengthening the second immunization was applied, and the immune effect was evaluated based on oocyst excretion, body weight gain, fecal scores, intestinal lesion scores and survival percentages. The results showed that no significant intestinal damage was caused by immunization reactions in the chickens. In addition, the number of excreted oocysts in the immunized chicken groups could be significantly increased, and the immunologic homogeneity of the immunized chickens could be improved by artificially strengthening the second immunization, which could in turn improve the immune protective effect.

  10. Attenuation of a drug-sensitive strain of a turkey protozoan parasite Eimeria meleagrimitis by selection for precocious development.

    Science.gov (United States)

    Rathinam, T; Gadde, U; Chapman, H D

    2016-01-30

    An attenuated line of Eimeria meleagrimitis was established by repeated propagation of the parasite in 9-day old turkey poults and subsequent selection for precocious development. Following 20 passages, the prepatent period decreased from 120 to 104h. A series of experiments were conducted to evaluate the pathogenicity, immunogenicity and fecundity of the newly selected line. Judged by body weight gain, feed consumption and feed efficiency following infection, the attenuated line had appreciably reduced pathogenicity. Immunogenicity of the attenuated line was examined by infecting poults successively with incremental doses of 10(2), 10(3) and 10(4) oocysts at 0, 7, and 14 days of age respectively. No oocysts were detected following challenge with 5×10(2) oocysts, indicating that the attenuated line had retained immunogenicity. Fecundity was assessed by infecting two-week old birds with 5×10(2) oocysts of either parent or attenuated line. Oocyst production from 96 to 240h post-infection showed that the patent period of the attenuated line commenced earlier and was of shorter duration than the parent line.

  11. Two new Eimeria species parasitic in corncrakes (Crex crex) (Gruiformes: Rallidae) in the United Kingdom.

    Science.gov (United States)

    Jeanes, C; Vaughan-Higgins, R; Green, R E; Sainsbury, A W; Marshall, R N; Blake, D P

    2013-08-01

    In this study we describe 2 new species of coccidia (Apicomplexa: Eimeriidae) parasites isolated from the feces of corncrake (Crex crex) (Gruiformes: Rallidae), bred in captivity in the U.K. Oocysts of Eimeria crecis n. sp. were approximately spherical and measured 15.3 μm (13-18) × 14.3 (12-16), providing an oocyst shape index of 1.1. A micropyle and oocyst residuum were absent, but a polar granule was present. Oocysts of Eimeria nenei n. sp. were ellipsoidal and measured 23.6 (21-26) × 18.1 (17-20), providing an oocyst shape index of 1.3. A micropyle and polar granule were present. Surveying free-living, wild adult corncrakes in Scotland (U.K.) demonstrated both parasite species to be widespread. These are the first species described to infect the corncrake, and they are distinct from those previously found to infect members of the closely related crane family (Gruiformes: Gruidae). Partial amplification and sequencing of the 18S rRNA gene and internal transcribed spacer 2 indicated a close relationship between the 2 proposed new species as a group distinct from the Eimeria species known to infect cranes. These newly identified parasite species have been associated with enteric disease in corncrakes being prepared for reproduction in captivity and reintroduction into England (U.K.).

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

    Science.gov (United States)

    Drozd, C; Schwartzbrod, J

    1996-04-01

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

  13. Occurrence of parasites on fruits and vegetables in Norway.

    Science.gov (United States)

    Robertson, L J; Gjerde, B

    2001-11-01

    Between August 1999 and January 2001, samples of various fruits and vegetables obtained within Norway were analyzed by published methods for parasite contamination. Neither Cyclospora oocysts nor Ascaris (or other helminth) eggs were detected on any of the samples examined for these parasites. However, of the 475 samples examined for Cryptosporidium oocysts and Giardia cysts, 29 (6%) were found to be positive. No samples were positive for both parasites. Of the 19 Cryptosporidium-positive samples. 5 (26%) were in lettuce, and 14 (74%) in mung bean sprouts. Of the 10 Giardia-positive samples, 2 (20%) were in dill, 2 (20%) in lettuce, 3 (30%) in mung bean sprouts, 1 (10%) in radish sprouts, and 2 (20%) in strawberries. Mung bean sprouts were significantly more likely to be contaminated with Cryptosporidium oocysts or Giardia cysts than the other fruits and vegetables. Concentrations of Cryptosporidium and Giardia detected were generally low (mean of approximately 3 [oo]cysts per 100 g produce). Although some of the contaminated produce was imported (the majority, if sprouted seeds are excluded), there was no association between imported produce and detection of parasites. Crvptosporidium oocysts and Giardia cysts were also detected in water samples concerned with field irrigation and production of bean sprouts within Norway. This is the first time that parasites have been detected on vegetables and fruit obtained in a highly developed. wealthy country, without there being an outbreak situation. These findings may have important implications for global food safety.

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

    DEFF Research Database (Denmark)

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

    2014-01-01

    Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp. contaminat......Protozoan parasites belonging to the genus Cryptosporidium are transmitted e.g. by food and water and may cause severe diarrhoea, dehydration, weight loss and malnutrition. Ingestion of 10 oocysts can lead to infection and pathogenic symptoms. Thus, to characterize Cryptosporidium spp....... contamination level of river water, irrigation water and lettuce, 10L of water and 16 lettuce samples were collected four times in the period of, September – October 2013, with weekly intervals from six sample sites in and around Kumasi, Ghana. Oocysts were purified from water by sedimentation for 2 x 48 hours...... to characterize Cryptosporidium spp. positive samples were done by PCR amplification and sequencing of the SSU rRNA, the HSP70 and the GP60 genes after. Cryptosporidium oocysts were found in 75% of the water samples and on 43% of the lettuce with concentrations of 53 – 3268 per 10 L water and 11 – 118 oocyst per...

  15. Simultaneous detection of the protozoan parasites Toxoplasma, Cryptosporidium and Giardia in food matrices and their persistence on basil leaves.

    Science.gov (United States)

    Hohweyer, Jeanne; Cazeaux, Catherine; Travaillé, Emmanuelle; Languet, Emilie; Dumètre, Aurélien; Aubert, Dominique; Terryn, Christine; Dubey, Jitender P; Azas, Nadine; Houssin, Maryline; Loïc, Favennec; Villena, Isabelle; La Carbona, Stéphanie

    2016-08-01

    Toxoplasma gondii, Cryptosporidium spp. and Giardia intestinalis are emerging pathogen parasites in the food domain. However, without standardized methods for their detection in food matrices, parasitic foodborne outbreaks remain neglected. In this study, a new immunomagnetic separation assay (IMS Toxo) targeting the oocyst's wall of T. gondii was developed using a specific purified monoclonal antibody. Performance of this IMS Toxo coupled to microscopic and qPCR analyses was evaluated in terms of limit of detection (LOD) and recovery rate (RR) on: i) simple matrix (LOD = 5 oocysts; RR between 5 and 56%); ii) raspberries and basil (LOD = 33 oocysts/g; RR between 0.2 and 35%). Finally, to simultaneously extract the three protozoa from these food matrices, T. gondii oocysts were directly concentrated (without IMS Toxo) from the supernatant of the IMS of Cryptosporidium and Giardia (oo)cysts. This strategy associated to qPCR detection led to LOD <1 to 3 (oo)cysts/g and RR between 2 and 35%. This procedure was coupled to RT-qPCR analyses and showed that the three protozoa persisted on the leaves of basil and remained viable following storage at 4 °C for 8 days. These data strengthen the need to consider these protozoa in food safety.

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

    Science.gov (United States)

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

    2012-11-06

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

  17. The epizootiology of Eimeria infections in commercial broiler chickens where anticoccidial drug programs were employed in six successive flocks to control coccidiosis.

    Science.gov (United States)

    Chapman, H D; Barta, J R; Hafeez, M A; Matsler, P; Rathinam, T; Raccoursier, M

    2016-08-01

    The course of natural Eimeria infections in 6 successive broiler flocks at a commercial farm comprising 4 houses, where different anticoccidial drug programs were employed, was studied by counting the number of oocysts in the litter at weekly intervals. The course of infection in all flocks followed a bell shaped curve in which oocyst numbers, initially low, increased to a peak ranging from 36 × 10(3) to 74 × 10(3) oocysts/g (OPG) of litter around 3 to 4 wk of age. Numbers subsequently declined to 3 × 10(3) to 15 × 10(3) OPG. Oocysts could be detected between flocks when birds were not present. Species of Eimeria identified included E. acervulina, E. maxima, and E. tenella Despite the presence of large numbers of oocysts in the litter, coccidial lesions were not observed in the intestines of the birds. The performance of broilers at the study site was comparable to that of other farms in the area where birds from the same settlement were reared to a similar age using the same drug programs. The results indicate the ubiquitous nature of Eimeria spp. infections in commercial broilers despite prophylactic medication. © 2016 Poultry Science Association Inc.

  18. Tomaculocystis corpulenta n. gen., n. sp. (Apicomplexa: Eugregarinorida) parasitizing the little yellow cockroach, Cariblatta lutea (Blattodea: Ectobiidae), in Alabama and Florida with recognition of Tomaculocystis cylindrosa n. comb. and Tomaculocystis mukundai n. comb. parasitizing ectobiid cockroaches in India.

    Science.gov (United States)

    Clopton, Richard E

    2015-02-01

    Tomaculocystis corpulenta n. gen., n. sp. (Apicomplexa: Eugregarinorida: Septatorina: Gregarinidae) is described from populations of the little yellow cockroach, Cariblatta lutea (Blattodea: Ectobiidae), established in laboratory culture from samples collected in Alabama and Florida. Tomaculocystis n. gen. are differentiated from other members of Gregarina by a markedly elliptoid gametocyst inside a persistent, lomentiform hyaline epicyst; developmental organization and growth of the spore tubes from gametocyst surface tumidi; and dehiscence by extrusion of non-chain forming oocysts through spore tubes that barely extend beyond the epicyst wall. Gregarina cylindrosa, Gregarina discocephala, and Gregarina mukundai are recognized as members of Tomaculocystis, and G. cylindrosa is recognized as the senior synonym of G. discocephala. Thus, Tomaculocystis cylindrosa n. comb. and Tomaculocystis mukundai n. comb. are formed. Species of Tomaculocystis are distinguished based on gamont deutomerite and oocyst shape and size. The oocysts of T. corpulenta are broadly dolioform, lack 4 polar knobs, and possess distinct, unique polar plates. Oocysts of all other known species in the genus are more oblong in shape, possess 4 polar knobs, and lack the distinct polar plates observed in the oocysts of T. corpulenta. Host utilization and geographic distribution among gregarine genera parasitizing the cockroach family Ectobiidae reveal a pattern of host-parasite specificity linking gregarine genera with ectobiidid subfamilies. Overall patterns suggest a hypothesis of European endemicy for Gamocystis, but hypotheses for the origin and radiation of Tomaculocystis or species of Gregarina infecting cockroaches are confounded by the cosmopolitan spread of pest cockroach species among humans.

  19. Some coccidial parasites of the lizard Amphisbaena alba (Reptilia: Amphisbaenia: Amphisbaenidae

    Directory of Open Access Journals (Sweden)

    Ralph Lainson

    2003-10-01

    Full Text Available Five parasites are described in the lizard Amphisbaena alba (Amphisbaenidae from the state of Pará, North Brazil. Mature oocysts of Choleoeimeria amphisbaenae n. sp., are passed already mature in the faeces. They are ellipsoidal-cylindrical, average 33.7 x 22.8 µm and are devoid of micropyle, oocyst residuum or polar body. The colourless wall is smooth and of 2 layers. The 4 dizoic sporocysts have no Stieda body and average 13 x 9.3 µm. Endogenous stages develop in the epithelial cells of the gall-bladder in the manner described for the genus and may cause extensive tissue damage. Sporulation of Isospora capanemaensis n. sp., is completed 3 days after the oocysts are voided in the faeces. They average 14.8 x 14.5 µm and have no micropyle, oocyst residuum or polar body. The 2 tetrazoic sporocysts are pear-shaped, average 8.6 x 6.6 and have an inconspicuous Stieda body. Endogenous development is in the epithelial cells of the ileum, and heavy infections cause considerable tissue destruction. Multisporocystic oocysts passed in the faeces of one A. alba possibly originated from an invertebrate host ingested by the lizard. A globidium-like cyst in the digestive tract of A. alba measured 105 x 85 µm and contained many hundreds of merozoites. A stained kidney smear of the same lizard revealed the presence of an unidentified parasite producing multinucleate cyst-like stages.

  20. Four new coccidia (Apicomplexia: Eimeriidae) from anoles (Lacertilia: Polychrotidae) in the Dominican Republic.

    Science.gov (United States)

    Cisper, G L; Huntington, C; Smith, D D; Powell, R; Parmerlee, J S; Lathrop, A

    1995-04-01

    Fecal samples from 25 Anolis armouri, 2 Anolis bahorucoensis, 48 Anolis cybotes, and 21 Anolis olssoni (Lacertilia: Polychrotidae) from southern Hispaniola were examined for coccidian oocysts. Two eimerians and 2 isosporans are herein described as new species. Sporulated oocysts of Eimeria schwartzi n. sp. from A. armouri are ellipsoidal, 22.7 (20.8-25.0) x 15.7 (14.6-17.7) microns, with spherical to subspherical sporocysts, 7.9 (6.2-9.4) x 7.4 (6.2-8.3) microns. Sporulated oocysts of Isospora reui n. sp. from A. bahorucoensis are spherical to subspherical, 18.2 (15.6-20.0) x 17.8 (15.6-19.8) microns, with ovoid sporocysts, 11.9 (10.4-12.7) x 8.5 (7.5-9.4) microns. Sporulated oocysts of Isospora hendersoni n. sp. from A. armouri and A. cybotes are spherical to subspherical, 23.2 (20.8-26.0) x 21.1 (18.4-23.9) microns, with ellipsoidal sporocysts, 14.7 (12.5-15.6) x 10.0 (9.2-11.4) microns. Sporulated oocysts of Eimeria avilae n. sp. from A. olssoni are cylindrical, 29.3 (26.0-33.3) x 15.9 (13.5-18.9) microns, with ellipsoidal sporocysts 10.2 (9.4-11.4) x 6.8 (5.2-8.0) microns.

  1. Isospora manchacensis n. sp., an intranuclear coccidian from the Louisiana ground skink, Scincella lateralis (Say, 1823) (Lacertilia: Scincidae).

    Science.gov (United States)

    Atkinson, C T; Ayala, S C

    1987-08-01

    Isospora manchacensis n. sp. is described from ground skinks, Scincella lateralis (Say, 1823) from Louisiana. Overall prevalence at 6 sites near Lake Ponchartrain was 43.1% (59/137) and ranged from 8% (1/13) to as high as 60% (6/10). Endogenous stages develop inside the nuclei of epithelial cells in the small intestine. Infected hypertrophic nuclei migrate from the basal lamina of the host cell to the luminal striated border. Oocysts in freshly passed fecal pellets usually contain a single contracted sporont that divides to form 2 sporoblasts. These undergo a brief pyramid stage followed by sporulation within 45-50 hr. Sporulated oocysts have a single-layered wall and measure 25.0 X 22.6 (20.0-28.9 X 18.6-26.0) micron. The lemon-shaped sporocysts measure 12.8 X 10.2 (11.1-15.2 X 9.0-11.0) micron and contain a Steida body, a spherical to oval substeida body, and a dispersed, granular sporocyst residuum. Prepatent periods in skinks fed 700 and 1,400 oocysts ranged from 24 to 32 days. Experimentally infected skinks produced large numbers of oocysts continuously during the 3-4 wk they were monitored after the onset of patency, but exhibited no signs of disease. Experimental doses of 200 oocysts failed to produce infections in skinks monitored for as long as 7 wk.

  2. Restoration of sensitivity to salinomycin in Eimeria following 5 flocks of broiler chickens reared in floor-pens using drug programs and vaccination to control coccidiosis.

    Science.gov (United States)

    Chapman, H D; Jeffers, T K

    2015-05-01

    Five successive flocks of broilers were reared in floor-pens and given different drug programs or were vaccinated against coccidiosis. Oocysts of Eimeria were isolated from the litter of pens during the fifth flock and their sensitivity to salinomycin (Sal) investigated by measuring new oocyst production following infection of medicated and unmedicated birds. Parasites obtained following 5 flocks given Sal were not well-controlled and it was concluded that they were partially resistant to the drug. Parasites obtained following 4 unmedicated flocks and one medicated flock were better controlled by Sal and it was concluded that in the absence of continuous medication there had been an improvement in drug efficacy. Sal almost completely suppressed oocyst production of isolates from treatments in which medication was followed by vaccination, indicating that when a drug program is followed by vaccination, restoration of sensitivity to Sal had occurred.

  3. Scanning electron microscopy of damage to the cecal mucosa of turkeys infected with Eimeria adenoides.

    Science.gov (United States)

    Bemrick, W J; Hammer, R F

    1979-01-01

    White Wrolstad turkeys were each inoculated with 100,000 Eimeria adenoides oocysts and killed on days 4-14 postinoculation. Tissue samples, obtained from 4 areas of the ceca comparable to areas examined in chickens infected with E. tenella in previous studies, were processed by a modification of the osmium-thiocarbo-hydrazide-osmium technique and examined with a scanning electron microscope. The pathologic situation found in turkeys was slightly different from that in the ceca of chickens infected with E. tenella. The mucosal lesions are most severe at the proximal end of an infected cecum. Surface disruption was far less severe than with cecal coccidiosis in chickens of the same age exposed to an equal number of infective oocysts. Rupture of the epithelial cell often caused the mucosal surface to present a honeycomb appearance. Some specific stages of the life cycle were identified, including schizonts and oocysts.

  4. Internal parasites of reptiles.

    Science.gov (United States)

    Raś-Noryńska, Małgorzata; Sokół, Rajmund

    2015-01-01

    Nowadays a growing number of exotic reptiles are kept as pets. The aim of this study was to determine the species of parasites found in reptile patients of veterinary practices in Poland. Fecal samples obtained from 76 lizards, 15 turtles and 10 snakes were examined by flotation method and direct smear stained with Lugol's iodine. In 63 samples (62.4%) the presence of parasite eggs and oocysts was revealed. Oocysts of Isospora spp. (from 33% to 100% of the samples, depending on the reptilian species) and Oxyurids eggs (10% to 75%) were predominant. In addition, isolated Eimeria spp. oocysts and Giardia intestinalis cysts were found, as well as Strongylus spp. and Hymenolepis spp. eggs. Pet reptiles are often infected with parasites, some of which are potentially dangerous to humans. A routine parasitological examination should be done in such animals.

  5. Gray wolf (Canis lupus) is a natural definitive host for Neospora caninum.

    Science.gov (United States)

    Dubey, J P; Jenkins, M C; Rajendran, C; Miska, K; Ferreira, L R; Martins, J; Kwok, O C H; Choudhary, S

    2011-09-27

    The gray wolf (Canis lupus) was found to be a new natural definitive host for Neospora caninum. Neospora-like oocysts were found microscopically in the feces of three of 73 wolves from Minnesota examined at necropsy. N. caninum-specific DNA was amplified from the oocysts of all three wolves. Oocysts from one wolf were infective for the gamma interferon gene knock out (KO) mice. Viable N. caninum (designated NcWolfUS1) was isolated in cell cultures seeded with tissue homogenate from the infected mouse. Typical thick walled tissue cysts were found in outbred mice inoculated with the parasite from the KO mouse. Tissue stages in mice stained positively with N. caninum-specific polyclonal antibodies. Our observation suggests that wolves may be an important link in the sylvatic cycle of N. caninum.

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

    Science.gov (United States)

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

    2012-01-01

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

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

    Science.gov (United States)

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

    2003-05-01

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

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

    DEFF Research Database (Denmark)

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

    2015-01-01

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

  9. Eimeria stiedae

    Science.gov (United States)

    Rose, M. Elaine

    1959-01-01

    Circulating antibodies to Eimeria stiedae have been demonstrated in rabbit serum using precipitation in gel and liquid media and complement fixation tests. Sera prepared by subcutaneous injections of antigen were similar to those obtained after oral infection with oocysts of E. stiedae. Antigens prepared from crushed oocysts and the exudate of infected bile ducts had similar properties, although a constituent of the bile duct antigen was absent or only present in very small amounts in the oocyst antigen. Antigens obtained on different days after infection from bile ducts showed differing precipitation patterns in gel; the maximum number of bands was given by antigens obtained 17–19 days after infection, antigens taken before and after this period reacted to give fewer precipitation bands. PMID:13653731

  10. Dietary supplementation of Artemisia annua to free range broilers and its effects on gastro-intestinal parasite infections

    DEFF Research Database (Denmark)

    Almeida, Gustavo Fonseca; Horsted, Klaus; Thamsborg, Stig Milan

    2012-01-01

    of the 12 groups (n=120) were monitored twice a week for excretion of coccidia oocysts (OPG) and gastrointestinal nematode eggs (EPG). At the end of the trial, when broilers were 74 days of age, 5 broilers per plot (n=60) were necropsied and their intestines opened for coccidia speciation and to account...... for the presence of gastrointestinal nematodes. In general, broilers from both genotypes coped well with coccidia infections as no deaths were reported. Broilers supplemented with A. annua showed a reduced number of excreted oocysts (67% less; p... of oocysts than males (45% more; pgastrointestinal nematodes were observed during routine EPG. However, during necropsy, juvenile larvae of Ascaridia galli were found in 73% of the broilers with no differences in respect...

  11. Eimeria auratae n. sp. (Apicomplexa: Eimeriidae) infecting the lizard Mabuya aurata in Saudi Arabia.

    Science.gov (United States)

    Alyousif, M S; AL-Rasheid, K A

    2001-03-01

    Eimeria auratae n. sp. was described from the gall bladder of the lizard Mabuya aurata collected at Al-Hofuf village, eastern region, Saudi Arabia. Morphology of sporulated as well as non-sporulated oocysts were studied. Sporulated oocysts were ellipsoidal 22-31.5x13.5-21.8 (27.7x18.5) microm with smooth brownish-yellow bilayered wall, 1.1 (0.9-1.3) microm. Micropyle, polar granule and oocyst residuum were absent. Sporocysts were ellipsoidal 10.5-12.8x7.5-9 (11.8x8.5) microm. Sporocyst residuum was present but Stieda body was absent. Sporozoites were crescent-shaped, blunt at one end and slightly tapered at the other. Eimeria species from Scincidae were compared.

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

    Science.gov (United States)

    Tram, Nguyen Thuy; Dalsgaard, Anders

    2014-12-01

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

  13. Occurrence of Cryptosporidium species in catfish (Clarias gariepinus) Harvested from two lakes and artificial ponds in Zaria, Northern Nigeria

    Institute of Scientific and Technical Information of China (English)

    Joy Cecilia Atawodi; Armayau Hamisu Bichi

    2013-01-01

    Objective: To determine the occurrence of Cryptosporidium species in catfish (Clarias gariepinus) harvested from two lakes and ponds in Zaria, Kaduna State, Nigeria.Methods:Catfish samples (n=200) from two lakes and (n=200) from two private fish ponds were collected and their gills and gastrointestinal tract samples were analysed for Cryptosporidium oocysts using modified Ziehl-Neelsen staining technique was used, followed by microscopy. Results: Oocysts of Cryptosporidium species were found in all the sampling sites with an overall positivity rate of 49.75%. A comparison between the lakes and ponds as sampling sites revealed a statistically significant (P<0.05) higher percentage of occurrence of Cryptosporidium oocyst from the lakes (39.25%) than from the ponds (10.50%). Conclusions: These findings document for the first time the natural occurrence ofCryptosporidium sp. in catfish intended for human consumption and therefore, underlines the need for public enlightenment to guard against any possible zoonotic transmission.

  14. Prevalence of subclinical coccidiosis in broiler farms in Turkey.

    Science.gov (United States)

    Karaer, Zafer; Guven, Esin; Akcay, Aytac; Kar, Sirri; Nalbantoglu, Serpil; Cakmak, Ayse

    2012-03-01

    The presence of Eimeria spp. oocysts in fecal samples collected from 1,108 broiler houses in six regions, representing about 12% of all broiler farms in Turkey, was studied using the modified McMaster method. The age of the chickens in the 1,108 pens varied from 1 to 50 days. Oocysts were found in 602 (54.3%) of these broiler houses, and the mean OPG (oocysts per gram of feces) in those samples was 36,498.7 (50-952,000). No indication of clinical coccidiosis or other clinically evident infection or wide mortality was encountered in any of the pens studied. Further study showed that the age of the chickens, the occurrence of diarrhea on the houses and the density of broiler breeding in the area correlated with subclinical coccidiosis prevalence.

  15. Effect of irradiation on the viability of Toxoplasma gondii cysts in tissues of mice and pigs

    Energy Technology Data Exchange (ETDEWEB)

    Dubey, J.P.; Brake, R.J.; Murrell, K.D.; Fayer, R.

    1986-03-01

    Muscles from tongue, heart, and limbs of 14 pigs inoculated orally with Toxoplasma gondii oocysts were irradiated with 10, 20, 25, and 30 krad of gamma (cesium-137 and cobalt-60) irradiation. Viability of T gondii cysts was assayed by feeding porcine muscles to T gondii-free cats and/or by inoculation of sediment from acid-pepsin digested porcine muscle into mice. Cats fed 500-g samples of muscles irradiated with up to 20 krad shed T gondii oocysts. Cats fed muscles irradiated with 25 or 30 krad did not shed oocysts. Mice were inoculated with 8 isolates of T gondii, and tissue cysts in their brains irradiated with up to 40 krad were infective to mice; however, there was a 10,000-fold reduction in the viability of organisms in tissue cysts irradiated with 40 krad, compared with that in nonirradiated cysts. At 50 krad of gamma irradiation, there were no detectable infective organisms in infected mouse brains.

  16. In vitro cultivation of Hammondia heydorni: Generation of tachyzoites, stage conversion into bradyzoites, and evaluation of serologic cross-reaction with Neospora caninum.

    Science.gov (United States)

    Gondim, L F P; Meyer, J; Peters, M; Rezende-Gondim, M M; Vrhovec, M G; Pantchev, N; Bauer, C; Conraths, F J; Schares, G

    2015-06-15

    Hammondia heydorni was in vitro isolated from oocysts shed by three dogs using a finite cell line from embryonal bovine heart (KH-R). The oocysts were purified and suspended in 2% potassium dichromate or 2% sulphuric acid for sporulation for 2-5 days at room temperature. The parasites were confirmed as H. heydorni by PCR using specific primers (JS4/JS5) and by negative reaction for Neospora caninum employing the primers Np6+/Np21+. H. heydorni sporulated oocysts (1 × 10(6)) from each dog were initially treated with sodium hypochlorite. For excystation of sporozoites, oocysts from one dog were lysed by ultrasound followed by incubation with 0.75% taurocholate. Excystation of sporozoites from the other two dogs was achieved by oocyst fragmentation with glass beads with no further chemical treatment. Tachyzoites were clearly seen in the cultures at three days post inoculation (dpi). Bradyzoite conversion and cyst formation were evaluated at different time points by using a polyclonal rabbit serum against a bradyzoite-specific antigen (anti-BAG1), and a rat monoclonal antibody (mAbCC2) against a cyst wall protein. Bradyzoites were firstly detected at 7 dpi. Between 18 and 21 dpi most of cultured parasites consisted of encysted bradyzoites. The H. heydorni cysts increased in size during cultivation and reached a length of up to 135 μm. The parasite was maintained in the bovine heart cells up to 4.5months. Sera from mice and sheep experimentally infected with H. heydorni oocysts reacted with H. heydorni by IFAT, but did not cross-react with N. caninum antigens using IFAT or immunoblot. These findings suggest that serological cross-reactivity between H. heydorni and N. caninum seems to be of minor importance.

  17. Three new species of Coccidia (Apicomplexa: Eimeriidae) from Skinks, Lipinia spp. (Sauria: Scincidae), from Oceania

    Science.gov (United States)

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

    2013-01-01

    Between September 1991 and March 1993, 25 moth skinks (Lipinia noctua) were collected from various localities on the Cook Islands, Fiji, Papua New Guinea (PNG), and Vanuatu and examined for coccidians. In addition, a single Roux's lipinia skink (Lipinia rouxi) was collected from PNG and examined for coccidia. Sixteen (64%) L. noctua were found to harbor 2 new eimerians, and L. rouxi harbored another new Eimeria sp. Oocysts of Eimeria lipinia n. sp. from 9 (36%) L. noctua from the Cook Islands, Fiji, and PNG were subspherical with a bilayered wall and measured (L × W) 18.6 × 16.9 μm, with a L/W ratio of 1.1. Both micropyle and oocyst residuum were absent, but a polar granule was present. Oocysts of Eimeria melanesia n. sp. from 6 (24%) L. noctua from Fiji and Vanuatu and a single L. rouxi from PNG were subspherical to ellipsoidal with a bilayered wall and measured 19.8 × 17.5 μm, and L/W was 1.1. Both micropyle and oocyst residuum were absent, but a single or fragmented polar granule was present. Oocysts of Eimeria lessoni n. sp. from 1 (4%) L. noctua from PNG were cylindroidal with a bilayered wall and measured 28.1 × 15.7 μm, and L/W was 1.8. Both micropyle and oocyst residuum were absent, but a single polar granule was present. These represent the third report of Eimeria spp. reported from any host on PNG and the only coccidians, to our knowledge, ever described from L. noctua and L. rouxi and from the Cook Islands and Vanuatu.

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

    Science.gov (United States)

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

    2011-01-01

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

  19. Disease surveillance of Atlantic herring: molecular characterization of hepatic coccidiosis and a morphological report of a novel intestinal coccidian

    Science.gov (United States)

    Friend, Sarah E; Lovey, J; Hershberger, Paul

    2016-01-01

    Surveillance for pathogens of Atlantic herring, including viral hemorrhagic septicemia virus (VHSV),Ichthyophonus hoferi, and hepatic and intestinal coccidians, was conducted from 2012 to 2016 in the NW Atlantic Ocean, New Jersey, USA. Neither VHSV nor I. hoferi was detected in any sample. Goussia clupearum was found in the livers of 40 to 78% of adult herring in varying parasite loads; however, associated pathological changes were negligible. Phylogenetic analysis based on small subunit 18S rRNA gene sequences placed G. clupearum most closely with other extraintestinal liver coccidia from the genus Calyptospora, though the G. clupearum isolates had a unique nucleotide insertion between 604 and 729 bp that did not occur in any other coccidian species. G. clupearum oocysts from Atlantic and Pacific herring were morphologically similar, though differences occurred in oocyst dimensions. Comparison of G. clupearum genetic sequences from Atlantic and Pacific herring revealed 4 nucleotide substitutions and 2 gaps in a 1749 bp region, indicating some divergence in the geographically separate populations. Pacific G. clupearum oocysts were not directly infective, suggesting that a heteroxenous life cycle is likely. Intestinal coccidiosis was described for the first time from juvenile and adult Atlantic herring. A novel intestinal coccidian species was detected based on morphological characteristics of exogenously sporulated oocysts. A unique feature in these oocysts was the presence of 3 long (15.1 ± 5.1 µm, mean ±SD) spiny projections on both ends of the oocyst. The novel morphology of this coccidian led us to tentatively name this parasite G. echinata n. sp.

  20. A study of the level and dynamics of Eimeria populations in naturally infected, grazing beef cattle at various stages of production in the Mid-Atlantic USA.

    Science.gov (United States)

    Lucas, Aaron S; Swecker, William S; Lindsay, David S; Scaglia, Guillermo; Neel, James P S; Elvinger, Francois C; Zajac, Anne M

    2014-05-28

    There is little information available on the species dynamics of eimerian parasites in grazing cattle in the central Appalachian region of the United States. Therefore, the objective of this study was to describe the level of infection and species dynamics of Eimeria spp. in grazing beef cattle of various age groups over the course of a year in the central Appalachian region. Rectal fecal samples were collected from male and female calves (n=72) monthly from May through October 2005, heifers only (n=36) monthly from November 2005 to April 2006, and cows (n=72) in May, July, and September, 2005. Eimeria spp. oocysts were seen in 399 of 414 (96%) fecal samples collected from the calves from May through October. Fecal oocysts counts (FOC) in the calves were lower (PEimeria spp. oocysts were detected in 198 of 213 (92%) of fecal samples collected from the 36 replacement heifers monthly from November to April and monthly mean FOC did not differ during this time period. The prevalence of oocyst shedding increased to 100% in calves in September and remained near 100% in the replacement heifers during the sampling period. Eimeria spp. oocysts were also detected in 150 of 200 (75%) samples collected in May, July, and September from the cows and mean FOC did not differ significantly over the sampling period. Eimeria spp. composition was dominated by Eimeria bovis in fecal samples collected from calves, replacement heifers and cows. Mixed Eimeria spp. infections were, however, common in all groups and 13 Eimeria spp. oocysts were identified throughout the sampling period.

  1. A low-virulence Eimeria intestinalis isolate from rabbit (Oryctolagus cuniculus) in China: molecular identification, pathogenicity, and immunogenicity.

    Science.gov (United States)

    Shi, Tuanyuan; Bao, Guolian; Fu, Yuan; Suo, Xun; Hao, Lili

    2014-03-01

    An Eimeria intestinalis isolated from a rabbit in China was first identified by amplifying the 18S small subunit (SSU) ribosomal RNA gene. The size of the amplified fragment was 1521 bp. The 18S SSU RNA gene of the E. intestinalis isolate shared 99% sequence identity with E. intestinalis isolates from France and the Czech Republic, with 100 and 96% coverage, respectively. Then, the pathogenicity and immunogenicity of the E. intestinalis isolate were evaluated in specific pathogen free (SPF) rabbits. In the pathogenicity assay, SPF rabbits in four groups were infected with 5 × 10(3), 5 × 10(4), 5 × 10(5), and 0 sporulated oocysts, respectively. Clinical signs including diarrhoea, constipation, loss of appetite, and reduction of body weight gain were observed in rabbits inoculated with 5 × 10(4) and 5 × 10(5) oocysts. And one rabbit (25 %) inoculated with 5 × 10(5) oocysts died 15 days after the inoculation. In the immunogenicity assay, SPF rabbits in five groups (named B1, B2, B3, B4, and B5) were immunised with 5 × 10(1), 5 × 10(2), 5 × 10(3), 0, and 0 sporulated oocysts, respectively. All rabbits but the B5 group were challenged with 1 × 10(6) oocysts. After the challenge, no or slight clinical signs were seen in rabbits of the B2 and B3 groups. Compared with the control, a 69.6 and 84.5% reduction of oocyst output was observed in the B2 and B3 groups, respectively. The body weight gain of the two groups was obviously higher than that of the challenge control group. All the results show that the E. intestinalis isolate has low virulence but immunogenicity in rabbit.

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

    Science.gov (United States)

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

    2007-05-01

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

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

    Science.gov (United States)

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

    2013-10-18

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

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

    Science.gov (United States)

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

    2014-06-15

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

  5. Disease surveillance of Atlantic herring: molecular characterization of hepatic coccidiosis and a morphological report of a novel intestinal coccidian.

    Science.gov (United States)

    Friend, Sarah E; Lovy, Jan; Hershberger, Paul K

    2016-07-07

    Surveillance for pathogens of Atlantic herring, including viral hemorrhagic septicemia virus (VHSV), Ichthyophonus hoferi, and hepatic and intestinal coccidians, was conducted from 2012 to 2016 in the NW Atlantic Ocean, New Jersey, USA. Neither VHSV nor I. hoferi was detected in any sample. Goussia clupearum was found in the livers of 40 to 78% of adult herring in varying parasite loads; however, associated pathological changes were negligible. Phylogenetic analysis based on small subunit 18S rRNA gene sequences placed G. clupearum most closely with other extraintestinal liver coccidia from the genus Calyptospora, though the G. clupearum isolates had a unique nucleotide insertion between 604 and 729 bp that did not occur in any other coccidian species. G. clupearum oocysts from Atlantic and Pacific herring were morphologically similar, though differences occurred in oocyst dimensions. Comparison of G. clupearum genetic sequences from Atlantic and Pacific herring revealed 4 nucleotide substitutions and 2 gaps in a 1749 bp region, indicating some divergence in the geographically separate populations. Pacific G. clupearum oocysts were not directly infective, suggesting that a heteroxenous life cycle is likely. Intestinal coccidiosis was described for the first time from juvenile and adult Atlantic herring. A novel intestinal coccidian species was detected based on morphological characteristics of exogenously sporulated oocysts. A unique feature in these oocysts was the presence of 3 long (15.1 ± 5.1 µm, mean ±SD) spiny projections on both ends of the oocyst. The novel morphology of this coccidian led us to tentatively name this parasite G. echinata n. sp.

  6. Experimental transmission of intestinal coccidiosis to piglets: clinical, parasitological and pathological findings.

    Science.gov (United States)

    Robinson, Y; Morin, M; Girard, C; Higgins, R

    1983-01-01

    Twenty-eight piglets coming from a "specific pathogen free" herd were inoculated at three days of age with 50 000 or 100 000 sporulated oocysts of Isospora suis. Fecal samples were examined for oocyst shedding daily and several clinical parameters were recorded. Ten piglets were used as normal controls. Groups of piglets were euthanized from three days to 12 days postinoculation and routine necropsies were performed. Bacteriological, virological, parasitological and histopathological examinations were made on the intestinal tracts. The incubation period was four to five days. Clinical signs and microscopic intestinal lesions observed in the experimentally infected animals were similar to those reported in spontaneous cases of porcine neonatal coccidiosis. Lesions of villous atrophy in the small intestine seemed to result from the destruction of villous epithelial cells mainly during the peak of asexual reproduction which occurred around four to five days postinoculation. Intracellular coccidial organisms were difficult to find during the late atrophic and villous regrowth stages of the intestinal lesions. The prepatent period varied from four to seven days and the most common was five days. Eighty percent of the piglets kept alive more than four days postinoculation have shed oocysts. Piglets dosed with old sporulated oocysts (ten months old) shed many more oocysts than those infected with a fresh inoculum (less than two months old). The patent period was not determined precisely with the design of the experiment but some of the infected piglets shed oocysts for at least five days. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 5. Fig. 6. PMID:6230142

  7. Renal infection by a new coccidian genus in big brown bats (Eptesicus fuscus).

    Science.gov (United States)

    Wünschmann, Arno; Wellehan, James F X; Armien, Anibal; Bemrick, William J; Barnes, Donald; Averbeck, Gary A; Roback, Richard; Schwabenlander, Marc; D'Almeida, Edgar; Joki, Ron; Childress, April L; Cortinas, Roberto; Gardiner, Chris H; Greiner, Ellis C

    2010-02-01

    A novel coccidian parasite from the kidney of big brown bats (Eptesicus fuscus) is described. This coccidian (Nephroisospora eptesici nov. gen., n. sp.) was associated with a generally mild, focal or multifocal, well-demarcated cortical renal lesion less than 1 mm in diameter. The lesion represented cystic, dilated tubules with hypertrophied tubular epithelial cells and was present in the kidneys of 29 of 590 bats. Numerous coccidian parasites in various stages of development were present within the tubular epithelial cells and within the cyst lumina. Oocysts were collected from cystic dilated tubules. Thin-walled, sporulated ellipsoidal oocysts measuring an average of 18.9 x 20.8 microm were present in kidney tissue. The oocysts contained 2 sporocysts with 4 sporozoites. A polar body and a prominent oocyst residuum were present in the oocysts, but no micropyle, sporocyst residuum, or Stieda bodies were detected. Analysis of the 18S rRNA gene sequence put the parasite in the Sarcocystidae. The parasite is closely related to Besnoitia, Hammondia, Neospora, and Toxoplasma. Ultrastructural features, such as the presence of an apical complex in merozoites, support the identification of a coccidian. A new genus and species, Nephroisospora eptesicii, is proposed for this unusual coccidian in which the entire cycle is completed in the kidney of a single host; it has a membrane-like oocyst wall, sporogony occurs in the host rather than in the abiotic environment, and the positioning of the parasite by nucleic acid sequence indicates it to be closely allied to Sarcocystis and Besnoitia.

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

    Science.gov (United States)

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

    2010-06-01

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

  9. Use of Artemisia annua as a natural coccidiostat in free-range broilers and its effects on infection dynamics and performance

    DEFF Research Database (Denmark)

    Almeida, Gustavo Fonseca; Horsted, Klaus; Thamsborg, Stig Milan

    2012-01-01

    This work investigated the preventive effect of Artemisia annua L. dried leaves supplied as a botanical coccidiostat to two broiler genotypes reared in a Danish free-range system in a factorial experiment (two genotypes and ± supplement of dried A. annua leaves). The genotypes White Bresse L40...... animals per plot. This infection strategy was meant to imitate the transmission pathway observed at farm level. Ten individual birds from each of the 12 groups, in total 120 animals of mixed sex, were monitored twice weekly for 30 days for oocysts excretion. PCR of pooled faecal samples, oocyst morphology...

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

    Science.gov (United States)

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

    2002-08-01

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

  11. Four New Species of Eimeria (Apicomplexa: Eimeriidae) from Emoia spp. Skinks (Sauria: Scincidae), from Papua New Guinea and the Insular Pacific.

    Science.gov (United States)

    McAllister, Chris T; Duszynski, Donald W; Austin, Christopher C; Fisher, Robert N

    2017-02-01

    Between September and November 1991, 54 adult skinks from 15 species were collected by hand or blowpipe from several localities on Rarotonga, Cook Islands, Ovalau Island, Fiji, and Papua New Guinea (PNG), and their feces were examined for coccidians. Species included 5 seaside skinks (Emoia atrocostata), 1 Pacific blue-tailed skink (Emoia caeroleocauda), 2 Fiji slender treeskinks (Emoia concolor), 15 white-bellied copper-striped skinks (Emoia cyanura), 1 Bulolo River forest skink (Emoia guttata), 6 dark-bellied copper-striped skinks (Emoia impar), 5 Papua five-striped skinks (Emoia jakati), 2 Papua slender treeskinks (Emoia kordoana), 3 Papua robust treeskinks (Emoia longicauda), 1 brown-backed forest skink (Emoia loveridgei), 3 Papua black-sided skinks (Emoia pallidiceps), 2 Papua white-spotted skinks (Emoia physicae), 2 Papua yellow-head skinks (Emoia popei), 1 Papua brown forest skink (Emoia submetallica), and 5 Fiji barred treeskinks (Emoia trossula) Species of Eimeria (Ei.) were detected from these Emoia (Em.) spp. and are described here as new. Oocysts of Eimeria iovai n. sp. from Em. pallidiceps from PNG were ellipsoidal with a bilayered wall (L × W) 26.5 × 18.1 μm, with a length/width ratio (L/W) of 1.1. Both micropyle and oocyst residuum were absent, but a fragmented polar granule was present. This eimerian also was found in Em. atrocostata from PNG. Oocysts of Eimeria kirkpatricki n. sp. from Em. atrocostata from PNG were ellipsoidal with a bilayered wall, 18.6 × 13.5 μm, L/W 1.4. A micropyle and oocyst residuum were absent, but a fragmented polar granule was present. This eimerian was also shared by Em. cyanura from the Cook Islands and Fiji, Em. impar from the Cook Islands, Em. loveridgei from PNG, Em. pallidiceps from PNG, Em. popei from PNG, and Em. submetallica from PNG. Oocysts of Eimeria stevejayuptoni n. sp. from Em. longicauda were subspheroidal to ellipsoidal with a bilayered wall, 18.7 × 16.6 μm, L/W 1.1. A micropyle and oocyst residuum

  12. Strategies to reduce severity of coccidia infections in organic poultry systems by the use of plant extracts

    DEFF Research Database (Denmark)

    Almeida, Gustavo Fonseca; Horsted, Klaus; Thamsborg, Stig Milan

    2012-01-01

    treatment combinations and compared to a group vaccinated with a commercial anti-coccidia vaccine. We investigated dried leaves of cassava and Artemisia annua mixed in the diet for pullets at different ages and ethanolic concentrate extracts of A. annua and A. vulgaris via drinking water supplemented before...... or after appearance of clinical symptoms of the disease. Performance attributes and oocyst excretion (oocysts per g faeces = OPG) were monitored weekly from 1 to 14 weeks of age. The supplementation of A. annua in the diet (3% inclusion based on feed weight) influenced negatively the growth rates...

  13. Diagnostic stages of the parasites of the Florida manatee, Trichechus manatus latirostris.

    Science.gov (United States)

    Bando, Monica; Larkin, Iskande V; Wright, Scott D; Greiner, Ellis C

    2014-02-01

    Limited information is available on diagnostic stages of parasites in Florida manatees (Trichechus manatus latirostris). We examined 67 fecal samples from captive and wild manatees to define the diagnostic stages of the parasite fauna known to occur in Florida manatees. Parasite eggs were freshly extracted ex utero from identified mature helminths and subsequently characterized, illustrated, and matched to those isolated from fecal samples. In addition, coccidian oocysts in the fecal samples were identified. These diagnostic stages included eggs from 5 species of trematodes (Chiorchis fabaceus, Chiorchis groschafti, Pulmonicola cochleotrema, Moniligerum blairi, and Nudacotyle undicola), 1 nematode (Heterocheilus tunicatus), and oocysts of 2 coccidians (Eimeria manatus and Eimeria nodulosa).

  14. Efficacy of halofuginone lactate against experimental cryptosporidiosis in goat neonates.

    Science.gov (United States)

    Petermann, Julie; Paraud, Carine; Pors, Isabelle; Chartier, Christophe

    2014-05-28

    Preliminary results obtained in calves, lambs and goat kids infected by Cryptosporidium sp. have indicated a partial prophylactic efficacy of halofuginone lactate when administered at 100 μg/kg body weight (BW). In this study, the efficacy of halofuginone lactate was evaluated in goat neonates experimentally inoculated with Cryptosporidium parvum oocysts per oral route. The trial consisted in 2 replicated experiments carried out successively at 2 months of interval. Twenty-two 2- to 4-day-old kids were experimentally inoculated once, 2-3 days after the arrival in premises, with 10(6)C. parvum oocysts per oral route and were allocated into 2 groups. Animals of group 1 acted as untreated control whereas animals of group 2 received halofuginone lactate for 10 days from the infection day to day 9 post-infection (DPI) at a daily oral dose rate of 100 μg/kg BW. Individual oocyst shedding was monitored by daily examination of faecal smears stained by carbol fuchsin and scored semi-quantitatively (0-5) until 19 DPI. Daily diarrhoea scores, weight gain and mortality were recorded. In the first experiment, oocyst excretion started 1 DPI in the control group, was highest on 4 DPI (mean score 3.6) and became undetectable from 16-19 DPI. In the treated group, oocyst shedding started 1 day later, showed lower scores compared to control on 4, 5, 6, 7 and 10 DPI and vanished from 16 to 19 DPI. No significant difference was seen for weight gains between groups. Five kids died in the control group compared to 1 kid in the treated group. In the second (replicated) experiment, oocyst excretion started 2 DPI in the control group, was highest on 4 DPI (mean score 4.5) and became undetectable 18 and 19 DPI. In the treated group, oocyst shedding started 2 days later, peaked on 13 DPI (mean score 2.3) and persisted until the end of the experiment. No significant difference was seen for weight gains between groups. Ten kids died in the control group compared to 3 kids in the treated group

  15. Eimeria peltocephali n. sp., (Apicomplexa:Eimeriidae from the Freshwater Turtle Peltocephalus dumerilianus (Chelonia:Pelomusidae and Eimeria molossi n. sp., from the Bat, Molossus ater (Mammalia:Chiroptera

    Directory of Open Access Journals (Sweden)

    Lainson R

    1998-01-01

    Full Text Available The oocyst is described of Eimeria peltocephali n.sp. from faeces of the freshwater turtle Peltocephalus dumerilianus from Barcelos, State of Amazonas, Brazil. Sporulation is exogenous and fully developed oocysts are elongate, ellipsoidal or cylindrical, frequently curved to a banana-shape, 54.4 x19.1 (37.5 - 68.7 x 18.7-20.0 µm, shape-index 2.8 (1.8 -3.9. The oocyst wall is a single thin, colourless layer about 1 µm thick, with no micropyle. There is a bulky oocyst residuum, at first spherical to ellipsoidal, 19 x 16 (16. 2 -26.2 x 16 - 21.5µm , but becoming dispersed on maturation. There are no polar bodies. The sporocysts, 19.1 x 6.8 ( 17.5 -21.2 x 6.2 -7.5 µm, shape- index 2.8 (2.3 -3.2, are usually disposed in pairs at each end of the oocyst, and bear an inconspicuous Stieda body in the form of a flat cap. The sporozoites are elongate and slightly curved around the residuum. No refractile bodies were seen. Eimeria molossi n.sp., is described from the molossid bat Molossus ater. Sporulation is exogenous and the mature oocysts are predominantly broadly ellipsoidal, 23.4 x 17.5 (18-30 x 15-22.5 µm, shape-index 1.3 (1-1.6. The oocyst wall is about 2 µm thick, and of three layers: an inner thin, colourless one and two outer layers which are thicker, yellowish-brown, prominently striated and in close apposition. There is no micropyle or oocyst residuum, but one and occasionally two polar bodies are usually present. Sporocysts are ellipsoidal, 10.2 x 7.5 (10-12.5 x 7.5 µm, shape-index 1.4 (1.3-1.7 with an inconspicuous Stieda body. Endogenous stages are described in the epithelial cells of the small intestine

  16. Caecal coccidiosis in commercial male turkeys

    Directory of Open Access Journals (Sweden)

    Giuseppe Sironi

    2010-01-01

    Full Text Available An outbreak of coccidiosis with high mortality is reported in 30-day-old commercial turkeys. Grossly, a severe typhlitis with a large fibrino-necrotic core was present. Large numbers of oocysts were observed in caecal smears. The location and the severity of the lesions and the oocyst morphology were strongly suggestive of Eimeria adenoeides infection. This species has already been reported in turkey flocks in Italy, but it is rarely responsible for clinical coccidiosis and severe lesions with high mortality. Other caecal parasitic infections are considered in differential diagnosis.

  17. Effect of heat killed Mycobacterium phlei on body weight gain and management of caecal coccidiosis in broiler chickens.

    Science.gov (United States)

    Bera, A K; Bhattacharya, D; Pan, D; Manna, B; Bandyopadhyay, S; Das, S K

    2010-10-01

    The objective of this study was to evaluate the immunotherapeutic potential of heat killed Mycobacterium phlei in broiler chicken against experimentally produced Eimeria tenella infection. The selected dose of E. tenella oocyst (5x10(3) sporulated oocysts per bird) was capable of producing a mild form of caecal coccidiosis as observed by significant difference in body weight gain, clinical findings and caecal lesion score. Heat killed M. phlei was fed orally at 10 mg per bird with sterile PBS vehicle at alternate day for four doses. Our study reveals that per day body weight gain was significantly (pcoccidiosis in broiler chicken. 2010 Elsevier Ltd. All rights reserved.

  18. Caecal coccidiosis in commercial male turkeys

    Directory of Open Access Journals (Sweden)

    Tiziana Rampin

    2006-01-01

    Full Text Available An outbreak of coccidiosis with high mortality is reported in 30-day-old commercial turkeys. Grossly, a severe typhlitis with a large fibrino-necrotic core was present. Large numbers of oocysts were observed in caecal smears. The location and the severity of the lesions and the oocyst morphology were strongly suggestive of Eimeria adenoeides infection. This species has already been reported in turkey flocks in Italy, but it is rarely responsible for clinical coccidiosis and severe lesions with high mortality. Other caecal parasitic infections are considered in differential diagnosis.

  19. 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.

  20. Relation between infection by gastrointestinal helminths and coccidia in the hematocrit values and weight gain in goats (Capra hircus from the breed Anglo Nubian during growth phase

    Directory of Open Access Journals (Sweden)

    Elizabeth Regina Rodrigues da Silva

    2012-11-01

    Full Text Available This study aimed to monitor the correlation between infections by gastrointestinal helminths and coccidia in the weight gain and hematocrit tests of goats from the breed Anglo Nubian, during growth phase, reared in the Metropolitan Region of Recife. The research was carried out between May and October 2011. Data analysis was performed through descriptive statistics techniques: absolute distributions, percentages, and Spearman correlation coefficient. The results found were infections by parasites, with a predominance of Strongyloidea eggs, besides oocysts of Eimeria spp. and also a significant correlation between the presence of Eimeria spp. oocysts and decrease in the weight gain and hematocrit value.

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

    Science.gov (United States)

    Nasser, Abidelfatah M

    2016-02-01

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

  2. Efficacy of different anticoccidials against experimental coccidiosis in large white turkeys.

    Science.gov (United States)

    Cabel, M C; Norton, R A; Yazwinski, T A; Waldroup, P W

    1991-02-01

    Two trials were conducted to compare the efficacy of currently approved anticoccidials for turkeys against challenge using a field isolate of mixed Eimeria species; E. adenoides, E. gallopavonis, and E. meleagrimitis. Poults in wire-floored cages were fed unmedicated diets from day-old to 3 wk of age. Diets were supplemented with either amprolium (AMP, 125 mg/kg), butynorate (BUT, 375 mg/kg), monensin (MON-60, 60 mg/kg; MON-100, 100 mg/kg), halofuginone (HAL; 3 mg/kg), zoalene (ZOA; 125 mg/kg), or sulfadimethoxine plus ormetoprim (SUL + ORM, 62.5 mg/kg and 37.5 mg/kg, respectively). After 2 days on the test diets, poults were individually weighed and inoculated with sporulated coccidial oocysts from the field isolate. Total fecal collections were obtained for Days 0 to 5 and 6 to 10 to estimate oocyst output. At 10 days postinoculation, the birds were individually weighed and killed to determine severity of intestinal lesions. The HAL and MON were most effective and AMP, ZOA, and SUL + ORM were least effective in maintaining weight and in reducing the severity of intestinal lesions. All the coccidiostats tested reduced oocyst passage, but poults fed HAL produced fewer oocysts. The results demonstrated differences in efficacy among anticoccidials with the more recently approved drugs providing the best protection against coccidiosis.

  3. Seroprevalence of Neospora caninum in free-range chickens (Gallus domesticus) from the Americas

    Science.gov (United States)

    Neospora caninum and Toxoplasma gondii are biologically and morphologically similar coccidians with canids as definitive hosts for N.caninum and felids for T. gondii. Feral chickens have been used as indicators of soil contamination with T. gondii oocysts because they feed from ground. In the presen...

  4. Clinical outcome and vertical transmission variability among canine Neospora caninum isolates in a pregnant mouse model of infection.

    Science.gov (United States)

    Dellarupe, Andrea; Regidor-Cerrillo, Javier; Jiménez-Ruiz, Elena; Schares, Gereon; Unzaga, Juan Manuel; Venturini, María Cecilia; Ortega-Mora, Luis M

    2014-03-01

    We compared the clinical outcome and vertical transmission of six canine Neospora caninum isolates using a pregnant BALB/c model. Four of the isolates were obtained from oocysts of naturally infected dogs (Nc-Ger2, Nc-Ger3, Nc-Ger6 and Nc-6Arg) and two were from diseased dogs with neurological signs (Nc-Bahia and Nc-Liv). The dams were inoculated with 2×106 tachyzoites of each isolate at day 7 of pregnancy. Morbidity, mortality and the antibody responses were evaluated in both the dams and the offspring, as was parasite transmission to the progeny. The mortality rates varied from 100% in Nc-Bahia and Nc-Liv-infected pups to 19% or less for those infected with the isolates from oocysts. The vertical transmission rates varied from 9 to 53% for N. caninum from oocysts, compared with 100% for the Nc-Liv and Nc-Bahia isolates. All dams showed specific IgG responses against tachyzoite and rNc-GRA7 antigens, confirming Neospora infection. The highest IgG levels were detected in mice inoculated with the Nc-Liv and Nc-Bahia isolates. These results demonstrate marked differences in virulence between the N. caninum isolates obtained from oocysts and neurologically affected dogs. This variability could help us to explain the differences in the outcome of the infection in definitive and intermediate hosts.

  5. Seroprevalence of Toxoplasma gondii and Neospora caninum in feral cats (Felis silvestris catus) in Majorca, Balearic Islands, Spain

    Science.gov (United States)

    Felids are important in the epidemiology of Toxoplasma gondii infection because they are the only hosts that can excrete environmentally-resistant oocysts. Antibodies to T. gondii and Neospora caninum were determined in serum samples from 59 feral cats (Felis silvestris catus) captured in baited tra...

  6. The Genus Cyclospora (Apicomplexa: Eimeriidae, with a description of Cyclospora schneideri n.sp. in the snake Anilius scytale scytale (Aniliidae from Amazonian Brazil: a review

    Directory of Open Access Journals (Sweden)

    Ralph Lainson

    2005-04-01

    Full Text Available A review is made of the recorded species of the coccidian genus Cyclospora and major events leading up to the discovery of C. cayetanensis, which is responsible for serious outbreaks of diarrhoea in man and is one of the aetiological agents of "traveller's diarrhoea". Humans appear to be the specific hosts, with the entire life-cycle in the intestine: to date there is no convincing evidence that the disease is a zoonosis. A description is given of oocysts and endogenous stages of C. schneideri n.sp., in the snake Anilius scytale scytale. Sporulation is exogenous and completed after about one week at 24-26º. Mature oocysts 19.8 × 16.6 (15.1 × 13.8-25.7 × 20.1, shape-index 1.2 (1.0-1.3: no oocyst residuum or polar bodies. Oocyst wall a single colourless, smooth layer with no micropyle: it is rapidly deformed or broken. Sporocysts 13.6 × 9.4 (11.3 × 8.3-15.1 × 9.9, shape-index 1.4 (1.2-1.5 with an inconspicuous Stieda body. Sporozoites 11-13 × 2.5-3. Endogenous stages are intracytoplasmic in the epithelial cells of the small intestine and with the characters of the Eimeriorina.

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

    Science.gov (United States)

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

  8. The red fox (Vulpes vulpes) and the arctic fox (Vulpes lagopus) are definitive hosts of Sarcocystis alces and Sarcocystis hjorti from moose (Alces alces).

    Science.gov (United States)

    Dahlgren, Stina S; Gjerde, Bjørn

    2010-09-01

    The aim of this study was to determine whether foxes might act as definitive hosts of Sarcocystis alces in moose. In 2 experiments, 6 silver foxes (Vulpes vulpes) and 6 blue foxes (Vulpes lagopus) were fed muscle tissue from moose containing numerous sarcocysts of S. alces, and euthanazed 7-28 days post-infection (p.i.). Intestinal mucosal scrapings and faecal samples were screened microscopically for Sarcocystis oocysts/sporocysts, which were identified to species by means of species-specific primers and sequence analysis targeting the ssu rRNA gene. All foxes in both experiments became infected with Sarcocystis; the oocysts were fully sporulated by 14 days p.i., containing sporocysts measuring 14-15 x 10 microm. Molecular identification revealed that the oocysts/sporocysts belonged to 2 species, S. alces and Sarcocystis hjorti, although sarcocysts of S. hjorti were only identified in moose subsequent to the infection of foxes. In the first experiment, all 8 foxes also became infected with a Hammondia sp. derived from moose, shedding unsporulated, subspherical oocysts, measuring 10-12 microm in diameter, from 6-7 days p.i. onwards. The study proved that canids (the red fox and arctic fox) are definitive hosts for S. alces and S. hjorti, as had been inferred from the phylogenetic position of these species.

  9. Schellackia (Apicomplexa: Eimeriidae of the brazilian tree-frog, Phrynohyas venulosa (Amphibia: Anura from Amazonian Brazil

    Directory of Open Access Journals (Sweden)

    I. Paperna

    1995-10-01

    Full Text Available Endogenous stages of a Schellackia species are described in histological sections of the intestine of the tree-frog, Phrynohyas venulosa, from North Brazil. Most oocysts sporulate within the epithelial cells of the gut, but a few were detected in the lamina propria.

  10. Characterization of IgG monoclonal antibody targeted to both tissue cyst and sporocyst walls of Toxoplasma gondii

    Science.gov (United States)

    Toxoplasma gondii infects approximately one third of the human population and animals habiting terrestrial and aquatic environments. Its environmentally resistant oocysts are excreted by felids, and the stage encysted in tissues (tissue cysts), are important in the horizontal transmission of T. gon...

  11. Toxoplasma gondii: epidemiology, feline clinical aspects, and prevention

    Science.gov (United States)

    Toxoplasma gondii is a parasite of birds and mammals. Cats are the only definitive host and thus the only source of infective oocysts, but other mammals and birds can develop tissue cysts. While feline infections are typically asymptomatic, infection during human pregnancy can cause severe disease i...

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

    Science.gov (United States)

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

    2015-01-01

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

  13. Isolation of Viable Toxoplasma gondii from Tissues and Feces of Cats from Addis Ababa, Ethiopia

    Science.gov (United States)

    Cats are important in the epidemiology of Toxoplasma gondii because they are the only hosts that excrete environmentally resistant oocysts in feces. In the present study, hearts, serum, and feces from 36 feral cats from Addis Ababa area, Ethiopia were examined for T. gondii infection. Antibodies to ...

  14. High prevalence of toxoplasmosis in cats from Egypt: isolation of viable Toxoplasma Gondii, tissue distribution, and isolate designation

    Science.gov (United States)

    Cats are important in the epidemiology of Toxoplasma gondii because they are the only hosts that excrete environmentally resistant oocysts in feces. In the present study, 158 feral cats from Giza, Egypt were examined for T. gondii infection. Antibodies to T. gondii were found in 97.4% with modified ...

  15. [Coccidies genus Eimeria as a bioindicator of radioactive pollution of the biocenose].

    Science.gov (United States)

    Pel'gunov, A N

    2006-01-01

    The data on coccidies of rodents were collected in Chernobil (1989-1991) and in the regions of radioactive pollution in the Bryansk region of Russia (1992-1999). The surface pollution of experimental plots was different and come from 0.11 to 11.8 MBq/m2. 2185 rodent were examined in all. Thirteen types of coccidies p. Eimeria were found out in 525 small animals. The analysis of changes in morphological characters and oocysts sporulation in dependence of the level of radioactive pollution of biocenose was carried out. It was found out that parametric signs (length, width and form index of oocysts) were independent from radioactive pollution. At the some time the radioactive pollution renders a significant influence on the nonparametric signs (different types of capsule deformation and internal texture of oocysts) and the process of sporulation. With the increase of radioactive pollution the part of nonsporulated oocycts increased and the quantity of oocysts, corresponding to the description of given type (normal), decreased. This dependence is well described by the equation of logarithmic regression, that allows to use this indexes in the bioindication of the radioactive pollution of the biocenose.

  16. Eimeria tenella infections in chicken: aspects of host-parasite interaction

    NARCIS (Netherlands)

    Jeurissen, S.H.M.; Janse, E.M.; Vermeulen, A.N.; Vervelde, L.

    1996-01-01

    Intestinal coccidiosis, caused by various species of Eimeria, has become an economically important disease of poultry and livestock throughout the world. Infection of chickens starts after ingestion of oocysts when sporozoites penetrate the epithelium of the villi. After passage through the lamina p

  17. TOXOPLASMOSIS IN CAPTIVE DOLPHINS (TURSIOPS TRUNCATUS) AND WALRUS (ODOBENUS ROSMRUS)

    Science.gov (United States)

    Toxoplasma gondii infection in marine mammals is intriguing and indicative of contamination of the ocean environment and coastal waters with oocysts. Toxoplasma gondii infection was detected in captive marine mammals at a seaquarium in Canada. Antibodies to T. gondii were found in all 7 bottlenose ...

  18. ISOLATION OF TOXOPLASMA GONDII FROM BOTTLENOSE DOLPHINS (TURSIOPS TRUNCATUS)

    Science.gov (United States)

    Toxoplasma gondii infection in marine mammals is intriguing and indicative of contamination of the ocean environment and coastal waters with oocysts. In previous serological surveys > 90% of bottlenose dolphins (Tursiops truncatus) from the coasts of Florida, South Carolina, and California had antib...

  19. Malaria parasites in mosquitoes: laboratory models, evolutionary temptation and the real world

    NARCIS (Netherlands)

    Boëte, C.H.J.J.

    2005-01-01

    A recent study describing the effect of Plasmodium berghei infection on some Anopheles gambiae immune genes demonstrates that P. berghei is responsible for the upregulation of several genes involved in the immune response that affect parasitic development differently during the ookinete-to-oocyst de

  20. ENHANCED DAPI STAINING FOR CRYPTOSPORIDIUM IN WATER SAMPLES

    Science.gov (United States)

    The U.S. Environmental Protection Agency's Method 1623 is used to detect and quantify the presence of {ital Cryptosporidium} spp. oocysts in water. The protocol consists of concentrating a sample, staining this concentrate with a fluorescent antibody, and examining the sample mi...

  1. Interlaboratory validation of an improved U.S. Food and Drug Administration method for detection of Cyclospora cayetanensis in produce using TaqMan real-time PCR

    Science.gov (United States)

    A collaborative validation study was performed to evaluate the performance of a new U.S. Food and Drug Administration method developed for detection of the protozoan parasite, Cyclospora cayetanensis, on cilantro and raspberries. The method includes a sample preparation step in which oocysts are re...

  2. Low prevalence of Neospora caninum and Toxoplasma gondii antibodies in dogs in Jilin, Henan and Anhui Provinces of the People’s Republic of China

    Science.gov (United States)

    Background: Neospora caninum and Toxoplasma gondii are important pathogens of worldwide distribution. N. caninum is a major cause of abortion in cattle and dogs are main reservoirs because they excrete the environmentally resistant oocysts. Toxoplasmosis is a worldwide zoonosis and dogs are consider...

  3. Immunohistochemical localization of sex hormone receptors in two Raillietina tapeworms.

    Science.gov (United States)

    Chen, L; Sun, Y M; Mu, L; Zeng, Y; Li, H Y; Yang, T H

    2017-03-08

    Sex hormone receptors play critical roles in development and reproduction. However, it is not known whether they exist in Raillietina tapeworms, and if they do, whether they have a similar function to that in vertebrates. We examined the immunohistochemical distributions of androgen receptors (ARs), estrogen receptors (ERs), and progesterone receptors (PRs) in the tissues of two tapeworm species: Raillietina echinobothrida and Raillietina tetragona. Immunopositive ARs were found in the entire reproductive system of R. echinobothrida, including the testes, ovaries, and oocysts, and weakly immunopositive ERs and PRs were found in the testes, ovaries, and oocysts. Immunopositive ARs were also found throughout the entire reproductive system of R. tetragona, including the testes, ovaries, and oocysts, and weakly immunopositive ERs were in the testes and oocysts; the PRs were distributed in an immunonegative manner. The results show that androgens and their receptors play critical roles in reproductive system development in the two tapeworms. The immunoreactivity and tissue localizations of the sex hormone receptors suggest that, in both species, they have similar functions as in vertebrates, and modulate reproduction.

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

    Science.gov (United States)

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

    2008-12-01

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

  5. Waterborne toxoplasmosis investigated and analysed under hydrogeological assessment: new data and perspectives for further research.

    Science.gov (United States)

    Vieira, Flávia Pereira; Alves, Maria da Glória; Martins, Livia Mattos; Rangel, Alba Lucínia Peixoto; Dubey, Jitender Prakash; Hill, Dolores; Bahia-Oliveira, Lilian Maria Garcia

    2015-11-01

    We present a set of data on human and chicken Toxoplasma gondii seroprevalence that was investigated and analysed in light of groundwater vulnerability information in an area endemic for waterborne toxoplasmosis in Brazil. Hydrogeological assessment was undertaken to select sites for water collection from wells for T. gondii oocyst testing and for collecting blood from free-range chickens and humans for anti-T. gondii serologic testing. Serologic testing of human specimens was done using conventional commercial tests and a sporozoite-specific embryogenesis-related protein (TgERP), which is able to differentiate whether infection resulted from tissue cysts or oocysts. Water specimens were negative for the presence of viable T. gondii oocysts. However, seroprevalence in free-range chickens was significantly associated with vulnerability of groundwater to surface contamination (p toxoplasmosis in light of groundwater vulnerability information associated with prevalence in humans estimated by oocyst antigens recognition have implications for the potential role of hydrogeological assessment in researching waterborne toxoplasmosis at a global scale.

  6. Experimental toxoplasmosis in rats induced orally with eleven strains of Toxoplasma gondii of seven genotypes: Tissue tropism, tissue cyst size, neural lesions, tissue cyst rupture without reactivation, and ocular lesions

    Science.gov (United States)

    The protozoan parasite Toxoplasma gondii is one of the most widely distributed and most successful microorganism. Of all warm blooded hosts, only cats can excrete the environmentally resistant stage, the oocyst. T. gondii manipulates rodent behavior so that infected rodents are losing fear of the ca...

  7. Traditional goat husbandry may substantially contribute to human toxoplasmosis exposure

    Science.gov (United States)

    Raising goats in settings that are highly contaminated with oocysts of Toxoplasma gondii may contribute significantly to human exposure to this zoonotic parasite. Increasing consumption of young goats in Romania, where goats are typically reared in backyards that are also home to cats (the definitiv...

  8. A novel dual promoter DNA vaccine induces CD8+ response against Toxoplasma gondii sporozoite specific surface protein “SporoSAG” through non-apoptotic cells

    Directory of Open Access Journals (Sweden)

    Sultan Gülçe İz

    2014-01-01

    The results of this study reveal the ability of SporoSAG protein to induce CD8+ T lymphocyte response for the first time. Overall, SporoSAG protein can be included to multivalant vaccine formulations in future studies to increase the protection in infections acquired through T. gondii oocysts.

  9. Gray wolf (Canis lupus) is a natural definitive host for Neospora caninum

    Science.gov (United States)

    The gray wolf (Canis lupus) was found to be a new natural definitive host for Neospora caninum. This finding is based on the recovery of Neospora-like oocysts from the feces of 3 of 73 wolves from Minnesota examined at necropsy, and on successful amplification of N. caninum-specific sequences from ...

  10. Metaphylactic effect of Diclazuril 0.25% in suckling beef calves, during a coccidiosis outbreak in extensive farming.

    Science.gov (United States)

    Romero, Jorge; Sanabria, Rodrigo; Travería, Gabriel; Di Paolo, Leandro; Peralta, Luis

    2013-03-31

    The weight gain performance and oocysts reduction, as response to the metaphylactic treatment with Diclazuril 0.25% at the start of a coccidiosis outbreak, was studied by a cases-controls transverse study. Fifty-eight suckling calves of approximately 90 days old were randomly selected from an infected beef herd on extensive farming. The calves were weighted and individual faecal samples were taken for oocyst per gram count (OPG). Out of those, 29 were drenched with 1 mg kg(-1) of Diclazuril in one oral dose (group T), since the other 29 remained as control group (group C). Samples for OPG and weights were measured again at days 7 and 21 after treatment, respectively. Later, the groups were divided (by the median) in higher or lower OPG counts, and were compared in the same way, in order to remove those without apparent infection (lower OPG counts). The faecal oocysts reduction reached 99.5% (p<0.0001), for the treated group. Along the three weeks of study, an increment of 2.65 kg in 21 days (125 g day(-1), p=0.036), was seen in treated group respect to controls, but this difference increased to 3.94 kg in 21 days (p<0.0001), when only the calves with higher OPG were taken into account. These results highlight the magnitude of the subclinical impact of coccidiosis, biased by the individual susceptibility to infection, which leads to get heavier infections and express higher oocysts output.

  11. Clinical observations, pathology, bioassay in mice and serological response at slaughter in pigs experimentally infected with Toxoplasma gondii

    DEFF Research Database (Denmark)

    Wingstrand, Anne; Lind, Peter; Haugegaard, J.;

    1997-01-01

    Experimental infections of a total of 47 pigs with tachyzoites of the Toxoplasma gondii RH-strain, tissue cysts of the SSI-119 and R92 strains as well as oocysts of the SSI-119 strain were performed to determine the sensitivity of an indirect IgG-ELISA, using tachyzoite lysate of the RH-strain as...

  12. A new coccidian from Acomys cahirinus Desmarest, 1819, from Evolution Canyon, Lower Nahal Oren, Mount Carmel, Israel.

    Science.gov (United States)

    Couch, L; Blaustein, L; Duszynski, D W; Shenbrot, G; Nevo, E

    1997-04-01

    In 1994, fresh fecal samples were collected and examined for coccidian parasites from 43 spiny mice (Acomys cahirinus) and from 60 wood mice (Apodemus mystacinus). The 2 genera of rodents inhabit an area in Lower Nahal Oren. Mount Carmel, Israel, known as Evolution Canyon, which consists of opposite-facing slopes that are geologically identical, but micro-climatically very different. Acomys cahirinus is found primarily on the warmer and drier south-facing slope (SFS), whereas A. mystacinus primarily inhabits the cooler and wetter north-facing slope (NFS). None of the samples from the A. mystacinus contained coccidia, but 6 of 43 (14%) A. cahirinus individuals were discharging eimerian oocysts that we describe herein as a new species. Five of the 6 positive samples were from the SFS. Sporulated oocysts are ovoidal to subspheroidal, 26.5 x 22.9 (21-29 x 19-26) microns, without a micropyle, but with an oocyst residuum of 1 to several large clear globules and a medium-sized refractile polar body; they contain lemon-shaped sporocysts, 10.4 x 8.1 (10 11 x 7-10) microns, with a sporocyst residuum and Stieda body, but no sub-/or parastieda body. Sporozoites lie side by side, completely filling oocysts; each contains a large posterior refractile body.

  13. Effect on performance of weanling alpacas following treatments against gastro-intestinal parasites.

    Science.gov (United States)

    Thomas, Susan M; Morgan, Eric R

    2013-11-15

    Nematodes and coccidia are common parasites of alpacas (Vicugna pacos), and important causes of disease in this increasingly popular livestock species. Endoparasitic infestation is thought to increase at times of natural or imposed stress, and antiparasitic treatments are often administered, although to date there is little evidence regarding their effect. Thirty-one alpaca juvenilles (cria) were divided into four groups at weaning, and received either no treatment as a control (C), fenbendazole anthelmintic (FB), toltrazuril coccidiostat (T), or both treatments (FBT). Body weights and faecal egg/oocyst counts were recorded weekly for six weeks following treatment. Although the prophylactic treatments decreased faecal egg/oocyst counts of the target organisms in the short term, there was no significant difference in egg/oocyst output over the course of the trial from animals given wormer, coccidiostat or both treatments. The group receiving anthelmintic only showed a significant reduction in live weight gain (LWG), with no significant difference in LWG between the other groups. At the conclusion of the trial, 'wormed only' alpacas weighed 3.3% less than at weaning, losing an average 1.3 kg over six weeks, whereas average LWG in the control group was 2.5 kg. Antiparasitics transiently reduced egg/oocyst output but results suggest that further investigation is required on the action of anthelmintics administered to alpaca cria at weaning and their effect on animal health and welfare.

  14. Severe intestinal coccidiosis in a newborn lion (Panthera leo)

    Science.gov (United States)

    Severe coccidiosis was found in sections of small intestine of a less than 2 day old lion (Panthera leo) born in captivity. Schizonts, merozoites, gamonts, and unsporulated oocysts were located in epithelial cells of ileum. Ultrastructural examination indicated that schizonts divided by schizogony. ...

  15. Eimeria pavoaegyptica sp. nov. (Apicomplexa: Eimeriidae in faeces of Indian peacocks, Pavo cristatus Linnaeus, 1758 (Galliformes: Phasianidae from Egypt

    Directory of Open Access Journals (Sweden)

    Ismail Saad El-Shahawy

    2010-12-01

    Full Text Available Coprological examination of 15 Indian peacocks, Pavo cristatus, revealed the presence of a coccidium species of the genus Eimeria, which apparently represents a previously undescribed species. Sporulation is exogenous and fully developed oocysts of Eimeria pavoaegyptica sp. nov. are ellipsoidal, with a dimension of 15 (13-16 × 12 (10-12.9 μm and with a shape index of 1.25 (1-1.3. The sporulated oocysts have no micropyle but enclose one large rectangular-shaped polar granule and an oocyst residuum. The oocysts have a distinct two-layered wall, which is ~1.7 μm thick. The outer layer has a smooth texture; it fills ~¾ of the total thickness and appears bicolored. The sporocysts are boat-shaped, of about 10 (9-11 × 4 (4-4.7 μm; their average shape-index is 2.5 μm with a small pointed Stieda body and a smooth, thin single-layered wall. No substieda body is detected. The sporocysts contain numerous, nearly uniform granular residua. The sporozoites are banana-shaped, 6 × 3 μm and each has two different-sized refractile bodies.

  16. Eimeria infections in goats in Southern Portugal.

    Science.gov (United States)

    Silva, Liliana Machado Ribeiro da; Vila-Viçosa, Maria João Martins; Nunes, Telmo; Taubert, Anja; Hermosilla, Carlos; Cortes, Helder Carola Espiguinha

    2014-01-01

    Coccidiosis caused by Eimeria species is a major form of intestinal infection affecting intensively and semi-intensively reared goats. The province of Alentejo is the main goat-producing area in Portugal. Therefore, all 15 Serpentina goat farms in Alentejo were analyzed regarding the occurrence and diversity of Eimeria species. Fecal samples obtained from 144 animals (52.1% dairy goats, 47.9% pre-pubertal goats) were examined using the modified McMaster technique to determine the number of oocysts per gram of feces. Eimeria spp. oocysts were present in 98.61% of the fecal samples and, overall, nine different Eimeria species were identified. The most prevalent species were E. ninakohlyakimovae (88%) and E. arloingi (85%), followed by E. alijevi (63%) and E. caprovina (63%). The average number of oocysts shed was significantly lower in dairy goats than in pre-adult animals. Astonishingly, no clinical signs of coccidiosis were observed in any of the animals examined, even though they were shedding high numbers of oocysts and were infected with highly pathogenic species. Thus, implementation of routine diagnostic investigation of the occurrence and diversity of caprine Eimeria species may be a useful tool for determination and better understanding of their potential economic impact on goat herds in southern Portugal.

  17. 两种鸡艾美耳球虫冰箱保存后的毒力观察%Observation on the Virulence of E.tenella and E.necatrix Preserved in Refrigerator

    Institute of Scientific and Technical Information of China (English)

    李秋明; 徐光勋; 沙晓菲; 雒凯; 孙剑

    2011-01-01

    Two strains of Eimeria oocysts , Eimeria tenella,E. Necatrix, were divided into two parts. One was preserved at 4 ℃ in refrigerator for 6 months, and then proliferated once. The other as fresh control strain was proliferated monthly. Pathoge-nicity experiments were performed. The results showed that the virulence of Eimeria (E. Tenella and E. Necatrix) oocysts weakened obviously 6 months later and it was not able to achieve the same level as fresh strain's if oocysts were proliferated once. The oocysts should be proliferated twice or above before used.%鸡艾美耳球虫(柔嫩艾美耳球虫和毒害艾美耳球虫)卵囊分成两部分处理,一部分常规冰箱保存6个月,到期后增殖1次;另一部分每月繁殖1次,为新鲜对照虫株.之后,进行致病性试验.结果显示:冰箱保存6个月后,虫株毒力明显下降,仅增殖1次不能达到新鲜虫株毒力水平.如果两种球虫卵囊冰箱保存6个月后,用于试验研究前需至少继代繁殖2次.

  18. Intestinal Coccidia (Apicomplexa: Eimeriidae of Brazilian Lizards. Eimeria carmelinoi n.sp., from Kentropyx calcarata and Acroeimeria paraensis n.sp. from Cnemidophorus lemniscatus lemniscatus (Lacertilia: Teiidae

    Directory of Open Access Journals (Sweden)

    Lainson Ralph

    2002-01-01

    Full Text Available Eimeria carmelinoi n.sp., is described in the teiid lizard Kentropyx calcarata Spix, 1825 from north Brazil. Oocysts subspherical to spherical, averaging 21.25 x 20.15 µm. Oocyst wall smooth, colourless and devoid of striae or micropyle. No polar body or conspicuous oocystic residuum, but frequently a small number of fine granules in Brownian movement. Sporocysts, averaging 10.1 x 9 µm, are without a Stieda body. Endogenous stages characteristic of the genus: intra-cytoplasmic, within the epithelial cells of the ileum and above the host cell nucleus. A re-description is given of a parasite previously described as Eimeria cnemidophori, in the teiid lizard Cnemidophorus lemniscatus lemniscatus. A study of the endogenous stages in the ileum necessitates renaming this coccidian as Acroeimeria cnemidophori (Carini, 1941 nov.comb., and suggests that Acroeimeria pintoi Lainson & Paperna, 1999 in the teiid Ameiva ameiva is a synonym of A. cnemidophori. A further intestinal coccidian, Acroeimeria paraensis n.sp. is described in C. l. lemniscatus, frequently as a mixed infection with A. cnemidophori. Mature oocysts, averaging 24.4 x 21.8 µm, have a single-layered, smooth, colourless wall with no micropyle or striae. No polar body, but the frequent presence of a small number of fine granules exhibiting Brownian movements. Sporocysts 9 x 8, without a Stieda body. Endogenous stages epicytoplasmic, characteristic of the genus, in the upper ileum. The importance of a study of the endogenous stages of eimeriid coccidia is discussed.

  19. Is sporozoite refractile body protein expression different in Eimeria acervulina sporozoites isolated from non-immune versus immune chickens?

    Science.gov (United States)

    A hallmark of Eimeria infection in avians is the establishment of immunity against clinical signs of coccidiosis. Resistant birds experience improved weight gain and feed conversion efficiency and lack intestinal lesions. Oocysts excretion is reduced, but not eliminated, in such immune chickens. ...

  20. A rapid method for determining salinomycin and monensin sensitivity in Eimeria tenella

    Science.gov (United States)

    Standard methods of determining the ionophore sensitivity of Eimeria rely on infecting chickens with an isolate or a mixture of Eimeria spp. oocysts in the presence of different anti-coccidial drugs. The purpose of this study was to develop a rapid in vitro method for assessing salinomycin and mone...

  1. Bioaccumulation of Toxoplasma and Cryptosporidium by the crustacean Gammarus fossarum: involvement in biomonitoring survey and trophic transfer

    Science.gov (United States)

    The protozoans Toxoplasma gondii and Cryptosporidium parvum are public health priorities and their oocysts can persist in environment for long time. They are present in various watercourses as recreational, surface, drinking, river, and seawater and could interact with organisms. To evaluate the cap...

  2. Seroprevalence of Toxoplasma gondii in free-living Amazon river dolphins (Inia geoffrensis) from central Amazon, Brazil

    Science.gov (United States)

    Toxoplasma gondii is an important pathogen in aquatic mammals and its presence in these animals may indicate water contamination of aquatic environment by oocysts. Serum samples from 95 dolphins from free-living Amazon River dolphins (Inia geoffrensis) from Sustainable Development Reserve Mamirauá (...

  3. Isolation of viable Toxoplasma gondii, molecular characterization, and seroprevalence in elk (Cervus canadensis) in Pennsylvania, USA

    Science.gov (United States)

    Toxoplasmosis is a worldwide zoonosis. The ingestion of uncooked/undercooked meat and consumption of water contaminated with Toxoplasma gondii oocysts excreted by felids are the main modes of transmission of this parasite. Thousands of wild cervids are hunted or killed in traffic accidents yearly bu...

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

    NARCIS (Netherlands)

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

    2007-01-01

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

  5. 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...

  6. Isolation and genetic characterization of toxoplasma gondii from striped dolphin (Stenella coeruleoalba) from Costa Rica

    Science.gov (United States)

    Toxoplasma gondii infection in marine mammals is of interest because of mortality and mode of transmission. It has been suggested that marine mammals become infected with T. gondii oocysts washed from land to the sea. We report the isolation and genetic characterization of viable T. gondii from a st...

  7. Coccidian parasites of intertidal fishes from Wales: systematics, development, and cytochemistry.

    Science.gov (United States)

    Davies, A J

    1978-02-01

    Examination of littoral fish Blennius pholis and Cottus bubalis caught at Aberystwyth and Porth Cwyfan, Wales, U.K. revealed 2 species of coccidia. Eimeria dingleyi sp. n. Oocysts spherical (16.1-19.2) to subspherical (13.9-14.2 X 18.8-20.0) micron, with thin walls; sporulation outside the host to produce ellipsoid sporocysts; endogenous phases in epithelial cells throughout intestine; 26 of 58 B. pholis infected. Eimeria variabilis (Théohan) Reichenow. Oocysts spherical (11.9-14.6) to subspherical (9.2-10.9 X 13.9-14.3) micron, sporulation in lining of pyloric ceca and rectum; previously unrecorded schizonts and gametocytes present; 21 of 25 C. bubalis infected. Electron microscopy revealed that the oocyst wall of E. variabilis consists of a thin membrane whereas the sporocyst wall is thick and 3-layered. Typical oocyst wall-forming bodies were absent from the macrogamete. Cytochemical tests on the endogenous stages of E. dingleyi and E. variabilis indicated that in general they resembled other coccidia in their chemical constitution.

  8. Eimeria tenella infections in chicken: aspects of host-parasite interaction

    NARCIS (Netherlands)

    Jeurissen, S.H.M.; Janse, E.M.; Vermeulen, A.N.; Vervelde, L.

    1996-01-01

    Intestinal coccidiosis, caused by various species of Eimeria, has become an economically important disease of poultry and livestock throughout the world. Infection of chickens starts after ingestion of oocysts when sporozoites penetrate the epithelium of the villi. After passage through the lamina

  9. It's a dirty job--A robust method for the purification and de novo genome assembly of Cryptosporidium from clinical material.

    Science.gov (United States)

    Andersson, Sofia; Sikora, Per; Karlberg, Maria L; Winiecka-Krusnell, Jadwiga; Alm, Erik; Beser, Jessica; Arrighi, Romanico B G

    2015-06-01

    We have developed a novel strategy for the purification of Cryptosporidium oocysts from clinical samples using IMS and PCR amplification of target DNA to facilitate uniform coverage genome sequencing and de novo assembly. Our procedure could also be used for other microbial pathogens from clinical specimens.

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

    Science.gov (United States)

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

    2011-09-01

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

  11. Biologic and genetic characteristics of Toxoplasma gondii isolates in free-range chickens from Nicaragua, Central America

    DEFF Research Database (Denmark)

    Dubey, J.P.; Sundar, N.; Pineda, N.

    2006-01-01

    The prevalence of Toxoplasma gondii in free-ranging chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii in 98 free-range chickens (Gallus domesticus) from Nicragua was determined. Antibodies to T. gondi...

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

    Directory of Open Access Journals (Sweden)

    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.

  13. 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.

  14. Disruption of Plasmodium sporozoite transmission by depletion of sporozoite invasion-associated protein 1.

    Science.gov (United States)

    Engelmann, Sabine; Silvie, Olivier; Matuschewski, Kai

    2009-04-01

    Accumulation of infectious Plasmodium sporozoites in Anopheles spp. salivary glands marks the final step of the complex development of the malaria parasite in the insect vector. Sporozoites are formed inside midgut-associated oocysts and actively egress into the mosquito hemocoel. Traversal of the salivary gland acinar cells correlates with the sporozoite's capacity to perform continuous gliding motility. Here, we characterized the cellular role of the Plasmodium berghei sporozoite invasion-associated protein 1 (SIAP-1). Intriguingly, SIAP-1 orthologs are found exclusively in apicomplexan hemoprotozoa, parasites that are transmitted by arthropod vectors, e.g., Plasmodium, Babesia, and Theileria species. By fluorescent tagging with mCherry, we show that SIAP-1 is expressed in oocyst-derived and salivary gland-associated sporozoites, where it accumulates at the apical tip. Targeted disruption of SIAP-1 does not affect sporozoite formation but causes a partial defect in sporozoite egress from oocysts and abolishes sporozoite colonization of mosquito salivary glands. Parasites with the siap-1(-) mutation are blocked in their capacity to perform continuous gliding motility. We propose that arthropod-transmitted apicomplexan parasites specifically express secretory factors, such as SIAP-1, that mediate efficient oocyst exit and migration to the salivary glands.

  15. Disruption of Plasmodium Sporozoite Transmission by Depletion of Sporozoite Invasion-Associated Protein 1▿ §

    Science.gov (United States)

    Engelmann, Sabine; Silvie, Olivier; Matuschewski, Kai

    2009-01-01

    Accumulation of infectious Plasmodium sporozoites in Anopheles spp. salivary glands marks the final step of the complex development of the malaria parasite in the insect vector. Sporozoites are formed inside midgut-associated oocysts and actively egress into the mosquito hemocoel. Traversal of the salivary gland acinar cells correlates with the sporozoite's capacity to perform continuous gliding motility. Here, we characterized the cellular role of the Plasmodium berghei sporozoite invasion-associated protein 1 (SIAP-1). Intriguingly, SIAP-1 orthologs are found exclusively in apicomplexan hemoprotozoa, parasites that are transmitted by arthropod vectors, e.g., Plasmodium, Babesia, and Theileria species. By fluorescent tagging with mCherry, we show that SIAP-1 is expressed in oocyst-derived and salivary gland-associated sporozoites, where it accumulates at the apical tip. Targeted disruption of SIAP-1 does not affect sporozoite formation but causes a partial defect in sporozoite egress from oocysts and abolishes sporozoite colonization of mosquito salivary glands. Parasites with the siap-1(−) mutation are blocked in their capacity to perform continuous gliding motility. We propose that arthropod-transmitted apicomplexan parasites specifically express secretory factors, such as SIAP-1, that mediate efficient oocyst exit and migration to the salivary glands. PMID:19181869

  16. Malaria, Leishmaniasis and Shistosomiasis Vector Ecology, Transmission, Immunology and Prophylaxis in Kenya

    Science.gov (United States)

    1994-01-27

    or eggs cab be used to establish the approximate age of an infection and may indicate whether th insect is a suitable host. Recognition of metacyclics...Andrysiak, and A.D. Brandling -Bennett. Identification of Malaria Species by ELISA in Sporozoite and Oocyst Infected Anopheles From Western Kenya

  17. Cryptosporidium en Giardia in Nederlandse zwembaden

    NARCIS (Netherlands)

    Schets FM; Engels GB; Leenen EJTM; MGB

    2003-01-01

    Swimming-pool associated outbreaks of cryptosporidiosis have been frequently reported in the UK and USA. Cryptosporidium oocysts could sometimes be detected in the pool water or the filter backwash water in cases where the source of the outbreak was confirmed. The occurrence of Cryptosporidium and G

  18. Emissie van Cryptosporidium en Giardia door landbouwhuisdieren

    NARCIS (Netherlands)

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

    1999-01-01

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

  19. Intestinal Parasites in the Critically Endangered Przewalski's Gazelle ( Procapra przewalskii ) in China, with the Description of a New Species of Eimeria (Apicomplexa: Eimeriidae).

    Science.gov (United States)

    Wang, Yunping; Du, Shouyang; Yang, Yanbin; Zhang, Xin; Duszynski, Donald W; Bian, Jianghui; Cao, Yifan

    2016-10-01

    We used a flotation technique to survey fecal samples from 27 Przewalski's gazelles ( Procapra przewalskii Büchner, 1891) for intestinal parasites. Samples were collected from the Qinghai Lake region, Qinghai-Tibet plateau, China, in April 2015. We report parasites belonging to two nematode (Nematodirus, Marshallagia) genera and one apicomplexan (Eimeria) genus; one of the latter was a new species from the critically endangered Przewalski's gazelle. Sporulated oocysts of the new eimerian were spheroidal; they had a thick, outer wall without a micropyle, but with a distinct polar granule and an oocyst residuum. Oocysts measured a mean of 18.6(±0.8)×17.3(±0.9) μm in length and width, with a mean 1.1(±0.1) length/width ratio and a spheroidal oocyst residuum that had a mean width of 2.0(±0.7) μm. Sporocysts were ovoid, measuring a mean of 9.4(±0.6)×5.5(±0.5) μm in length and width, with a mean 1.7(±0.2) length/width ratio. A Stieda body and sporocyst residuum were present, but a substieda body was absent. Sporozoites each had a small anterior refractile body and a larger, posterior refractile body; a small nucleus was visible between them.

  20. Transmission of Hepatozoon americanum (Apicomplexa: Adeleorina) by ixodids (Acari: Ixodidae).

    Science.gov (United States)

    Ewing, S A; Mathew, J S; Panciera, R J

    2002-07-01

    American canine hepatozoonosis (ACH) caused by Hepatozoon americanum Vincent-Johnson, Macintire, Lindsay, Lenz, Baneth, and Shkap is an emerging, often fatal, tick-borne protozoal disease of dogs in the United States of America. Dogs acquire infection by ingesting ticks that contain oocysts. To understand the invertebrate (definitive) host range of H. americanum, experiments were carried out using four ixodids, Rhipicephalus sanguineus (Latreille), Dermacentor variabilis Say, Amblyomma americanum (L.), and Amblyomma maculatum Koch. Laboratory-reared nymphal ticks were fed on dogs that were either naturally or experimentally infected with H. americanum; when these ticks molted to the adult stage they were either fed to susceptible dogs or were dissected and examined for the presence of oocysts. Mature H. americanum oocysts were found in >90% of A. maculatum (both males and females), whereas oocysts were not found in any of the other three species. These results confirm that A. maculatum is an excellent host and vector for H. americanum and also suggest that this apicomplexan may have a narrow invertebrate host range, at least among ixodid ticks that are likely candidate vectors in the United States.

  1. 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...

  2. Renal coccidiosis in interior Canada geese, Branta canadensis interior Todd, of the Mississippi Valley population

    Science.gov (United States)

    Tuggle, Benjamin N.; Crites, John L.

    1984-01-01

    Kidneys from 309 Interior Canada geese from three locations in the Mississippi Flyway were examined for renal coccidia. Oocysts and/or young zygotes of Eimeria sp. were found in 6.8% of goose kidneys sampled. Only one type of renal coccidian oocyst was observed. Significantly more immature geese were infected than adults; however, there was no significant difference observed between the prevalences of infection in male and female birds. A host cellular response to zygotes and oocysts was noted in the majority of infected adult geese. Heavily infected kidneys were hypertrophic with minute foci on the surface of the organ. Histological examinations showed large numbers of unsporulated oocysts accumulated in distended collecting tubules, resulting in pressure necrosis to adjacent tissue and urate retention. Zygotes were observed in the cytoplasm of tubule cells and extracellularly in interstitial tissue. Infected tubule cells were characterized by the peripheral location of the nuclei, cytoplasmic basophilia, and cellular hypertrophy. This is the first report of an Eimeria sp. in the kidneys of Canada geese of the Mississippi Valley population.

  3. Comparison of control methods for coccidiosis in native Spanish Castellana Negra chickens

    Energy Technology Data Exchange (ETDEWEB)

    Miguel, J. A.; Asenjo, B.; Ciria, J.; Cacho, E. del; Calvo, J. L.

    2008-07-01

    Coccidiosis is a disease responsible for serious economic losses in the poultry industry. This paper compares the effect of coccidiosis infection in a population of experimentally infected Castellana Negra chickens previously administered the ionophores antibiotic monensin (Treatment 1), Alquernat Zyox, a herb-based product (Treatment 2), or a live vaccine based on oocysts selected for precocity (Treatment 3). Fifty birds per treatment were housed in captivity and weighed individually once every two weeks. At nine weeks they were infected with pathogenic oocysts of Eimeria tenella, E. acervulina and E. maxima. No significant differences (P<0.05) were seen in body weight between the birds in the three treatment groups after week 10. The average daily weight gain of the Alquernat Zycox-treated and vaccinated birds was similar over the entire experimental period, and more regular than that of the monensin-treated birds. The number of oocysts eliminated in the faeces and the degree of intestinal injury caused were analysed at 10, 11 and 12 weeks. The vaccine-treated birds shed a smaller number of oocysts in their faeces at 11 and 12 weeks than did those treated with monensin or Alquernat Zycox (P<0.001). At 11 and 12 weeks the vaccine- and Alquernat Zycox-treated birds showed significantly (P<0.05) less intestinal injury than the monensin-treated birds. (Author) 22 refs.

  4. Secondary transmission of cryptosporidiosis associated with well water consumption: two case studies.

    Science.gov (United States)

    Rezende, Natania Carol Cavalcante; Bezerra, Camila Loredana Pereira Alves Madeira; Almeida, Jéssica Jacinto Salviano de; Fernandes, Tatiane Uetti Gomes; Luz, Kleber Giovanni

    2016-04-01

    Cryptosporidiosis is a very prominent disease in the field of public health, and usually causes diarrhea. We describe two immunocompetent patients who presented with chronic diarrhea that was ultimately found to be caused by continuous exposure to well water contaminated with the microbial cysts (oocysts) of the Cryptosporidium spp parasite. We describe the patients' histories and possible explanations for their prolonged symptoms.

  5. Cryptosporidiosis in a black bear in Virginia.

    Science.gov (United States)

    Duncan, R B; Caudell, D; Lindsay, D S; Moll, H D

    1999-04-01

    Cryptosporidiosis has not been previously reported in black bears in North America, either free-roaming or captive. However, oocysts have been documented in two captive Malayan sun bears (Helarctos malayanus) located in zoological parks in Taiwan. Developmental stages of Cryptosporidium parvum were observed in tissue sections from the small intestine of a black bear cub found dead in Virginia (USA).

  6. Effect of the granulometric characteristics of monensin sodium on controlling experimental coccidiosis in broiler chickens.

    Science.gov (United States)

    Rambozzi, Luisa; Renna, Manuela; Cornale, Paolo; Perona, Giovanni; Malfatto, Vanda; Mimosi, Antonio

    2012-01-01

    The aim of the present study was to investigate the efficacy of monensin sodium grain size on controlling coccidiosis in broiler chickens. Three hundred and fifty Cobb 700 chicks were infected experimentally with 8 × 10(4) sporulated oocysts of Eimeria spp. The birds were fed with diets supplemented with powdered (n = 150) or granulated (n = 150) monensin, with particle sizes of ≤ 100 µm and 450-650 µm, respectively. The remaining chicks (n = 50) were used as an unmedicated control group. The birds' weights and number of oocysts per gram of litter in each group were recorded weekly. A total of 96 birds were randomly selected and culled during the trial. Their intestinal oocyst counts and lesion scores were determined. No significant differences in body weights were found between the groups at the end of the study. The percentages of infected animals, oocyst counts and lesion scores were significantly higher in the control group than in the other two groups; the two treated groups did not show any appreciable differences to each other. Enteritis was observed in 14 birds, all in the group supplemented with granulated monensin. These results suggest that powdered and granulated monensin have comparable efficacy in controlling coccidiosis in broiler chickens. However, treatments with monensin sodium of high grain size may be subject to contraindications relating to enteritis.

  7. Development of an in vitro assay for monensin and salinomycin sensitivity in Eimeria tenella

    Science.gov (United States)

    Applying ionophore drugs and synthetic chemicals to poultry feed has been used for over 50 years to control avian coccidiosis. While this strategy has been and continues to be effective, drug resistance has prompted alternatives such as drug rotation or vaccination with live Eimeria oocysts. Often...

  8. Occurrence of Toxoplasma gondii antibodies in birds from the Atlantic Forest, state of São Paulo, Brazil

    Science.gov (United States)

    Avians are considered important intermediate hosts for Toxoplasma gondii because they serve as source of infection for Felidae, which shed environmentally resistant oocysts after ingesting infected tissues. Little is known of the epidemiology of toxoplasmosis in wild birds. In the present study, ant...

  9. Interaction forces drive the environmental transmission of pathogenic protozoa.

    Science.gov (United States)

    Dumètre, Aurélien; Aubert, Dominique; Puech, Pierre-Henri; Hohweyer, Jeanne; Azas, Nadine; Villena, Isabelle

    2012-02-01

    The protozoan parasites Giardia duodenalis, Cryptosporidium spp., and Toxoplasma gondii are pathogens that are resistant to a number of environmental factors and pose significant risks to public health worldwide. Their environmental transmission is closely governed by the physicochemical properties of their cysts (Giardia) and oocysts (Cryptosporidium and Toxoplasma), allowing their transport, retention, and survival for months in water, soil, vegetables, and mollusks, which are the main reservoirs for human infection. Importantly, the cyst/oocyst wall plays a key role in that regard by exhibiting a complex polymeric coverage that determines the charge and hydrophobic characteristics of parasites' surfaces. Interaction forces between parasites and other environmental particles may be, in a first approximation, evaluated following the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory of colloidal stability. However, due to the molecular topography and nano- to microstructure of the cyst/oocyst surface, non-DVLO hydrophobic forces together with additional steric attractive and/or repulsive forces may play a pivotal role in controlling the parasite behavior when the organism is subjected to various external conditions. Here, we review several parameters that enhance or hinder the adhesion of parasites to other particles and surfaces and address the role of fast-emerging techniques for mapping the cyst/oocyst surface, e.g., by measuring its topology and the generated interaction forces at the nano- to microscale. We discuss why characterizing these interactions could be a crucial step for managing the environmental matrices at risk of microbial pollution.

  10. An experimental Toxoplasma gondii dose response challenge model to study therapeutic or vaccine efficacy in cats

    NARCIS (Netherlands)

    Cornelissen, J.B.W.J.; Giessen, van der J.W.B.; Takumi, K.; Teunis, P.F.M.; Wisselink, H.J.

    2014-01-01

    High numbers of Toxoplasma gondii oocysts in the environment are a risk factor to humans. The environmental contamination might be reduced by vaccinating the definitive host, cats. An experimental challenge model is necessary to quantitatively assess the efficacy of a vaccine or drug treatment. Prev

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

    Science.gov (United States)

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

    2003-01-01

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

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

    Science.gov (United States)

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

    2012-11-01

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

  13. Coccidial and helminth infections in goats kept indoors in the Netherlands

    NARCIS (Netherlands)

    Borgsteede, F.H.M.; Dercksen, D.P.

    1996-01-01

    An investigation was carried out on coccidial and helminth infections in goats kept indoors on five farms in the Netherlands. The goats were individually sampled. Coccidial oocysts were identified and nematode eggs counted. Larval cultures were made and infective larvae identified to the generic or

  14. Intestinal coccidia (Apicomplexa: Eimeriidae) of Brazilian lizards. Eimeria carmelinoi n.sp., from Kentropyx calcarata and Acroeimeria paraensis n.sp. from Cnemidophorus lemniscatus lemniscatus (Lacertilia: Teiidae).

    Science.gov (United States)

    Lainson, Ralph

    2002-03-01

    Eimeria carmelinoi n.sp., is described in the teiid lizard Kentropyx calcarata Spix, 1825 from north Brazil. Oocysts subspherical to spherical, averaging 21.25 x 20.15 micro m. Oocyst wall smooth, colourless and devoid of striae or micropyle. No polar body or conspicuous oocystic residuum, but frequently a small number of fine granules in Brownian movement. Sporocysts, averaging 10.1 x 9 microm, are without a Stieda body. Endogenous stages characteristic of the genus: intra-cytoplasmic, within the epithelial cells of the ileum and above the host cell nucleus. A re-description is given of a parasite previously described as Eimeria cnemidophori, in the teiid lizard Cnemidophorus lemniscatus lemniscatus. A study of the endogenous stages in the ileum necessitates renaming this coccidian as Acroeimeria cnemidophori (Carini, 1941) nov.comb., and suggests that Acroeimeria pintoi Lainson & Paperna, 1999 in the teiid Ameiva ameiva is a synonym of A. cnemidophori. A further intestinal coccidian, Acroeimeria paraensis n.sp. is described in C. l. lemniscatus, frequently as a mixed infection with A. cnemidophori. Mature oocysts, averaging 24.4 x 21.8 microm, have a single-layered, smooth, colourless wall with no micropyle or striae. No polar body, but the frequent presence of a small number of fine granules exhibiting Brownian movements. Sporocysts 9 x 8, without a Stieda body. Endogenous stages epicytoplasmic, characteristic of the genus, in the upper ileum. The importance of a study of the endogenous stages of eimeriid coccidia is discussed.

  15. Isolation and genetic characterization of viable Toxoplasma gondii from tissues and feces of cats from the central region of China

    Science.gov (United States)

    Cats are important in the epidemiology of toxoplasmosis because they are the only definitive hosts that excrete environmentally resistant Toxoplasma gondii oocysts. Little is known of feline toxoplasmosis in China and most of the literature is in Chinese. Here we summarized all published reports on ...

  16. Tyzzeria boae n. sp., (Apicomplexa: Eimeriidae, a New Coccidium from the Kidney of the snake Boa constrictor constrictor (Serpentes: Boidae

    Directory of Open Access Journals (Sweden)

    Ralph Lainson

    1994-12-01

    Full Text Available A new species of Tyzzeria is described from the kidney of the snake Boa constrictor constrictor Linnaeus, from the State of Pará, north Brazil. Oocysts from the coacal contents matured in eight days, at approximately 24°C. They measured 19.0 x 18.0 (15.0 x 15.0 - 22.5 x 21.5 µm, shape-index (length/width 1.0 (1.0 - 1.1. The oocyst wall is of an extremely delicate single, colourless layer, with no micropyle. Division of the oocyst contents into the 8 naked sporozoites leaves a bulky, spherical oocyst residuum averaging 15.5 x 14.8 (13.5 x 13.5 - 18.5 x 17.5 µm; the sporozoites measure an average of 11.0 x 1.8 (8.5 x 1.25 - 12.5 x 2.0 µm, and possess both anterior and posterior refractile bodies. Tyzzeria boae n.sp. is unique among the recorded species of the genus by virtue of its development in the epithelial cells of the distal convoluted tubules and collecting tubules of the kidney: stages in the merogony and gametogony of the parasite are described and figured.

  17. Genotypic Characterization of Cryptosporidium hominis from Water Samples in São Paulo, Brazil

    Science.gov (United States)

    Araújo, Ronalda S.; Dropa, Milena; Fernandes, Licia N.; Carvalho, Terezinha T.; Sato, Maria Inês Z.; Soares, Rodrigo M.; Matté, Glavur R.; Matté, Maria Helena

    2011-01-01

    The protozoan parasite Cryptosporidium has emerged as one of the most important water contaminants, causing waterborne outbreaks of diarrheal diseases worldwide. The small size of oocysts under the microscope and the possibility of changes in characteristics of oocysts, mainly in environmental samples, make the taxonomy of the genus difficult if morphologic characteristics are considered. This limitation encouraged the application of molecular methods to identify this microorganism. The aim of this study was to detect and identify by nested-polymerase chain reaction oocysts of Cryptosporidium present in water samples in the state of São Paulo, Brazil. Water samples were concentrated through a membrane filter, DNA was extracted by using a standard technique, and both amplification reactions used forward and reverse oligonucleotides that were complementary to Cryptosporidium 18S ribosomal RNA gene sequences. Thirty water samples from different sites of collection in the state of São Paulo were evaluated. Cryptosporidium oocysts were detected in 30% of the samples. By genoptyping, C. hominis and Cryptosporidium sp. were identified in recreational water and C. meleagridis was identified in surface water samples. This is the first report of C. hominis in environmental samples in Brazil. Although identification of Cryptosporidium is still a difficult task, molecular methods are essential for specific identification and are a helpful tool to aid to understand the epidemiology of this parasite in Brazil. PMID:22049036

  18. Use of Artemisia annua as a natural coccidiostat in free-range broilers and its effects on infection dynamics and performance

    DEFF Research Database (Denmark)

    Almeida, Gustavo Fonseca; Horsted, Klaus; Thamsborg, Stig Milan

    2012-01-01

    combination. The paddocks were cultivated with a mix of grass and clover. A separate group of broilers was naturally infected with Eimeria spp. oocysts and five animals nominated as “seeders” were introduced to the above mentioned 12 groups, 10 days after its formation, with each group consisting of 35...

  19. Browse Title Index

    African Journals Online (AJOL)

    Items 501 - 550 of 683 ... Vol 53, No 1 (2005), Replacement of groundnut cake and maize with ... supplementation of Urea-Ammoniated rice straw of untreated rice straw fed ... indica) and garlic (Allium sativum) on oocyst count, bacteria count and ...

  20. 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...

  1. A new coccidian parasite, Isospora samoaensis, from the Wattled Honeyeater (Foulehaio carunculata) from American Samoa

    Science.gov (United States)

    Adamczyk, Kelly J.; McQuistion, Thomas E.; LaPointe, Dennis

    2004-01-01

    A new species of Isospora is described from the feces of the wattled honeyeater, Foulehaio carunculata from American Samoa. Numerous oocysts of similar morphology were found in a single adult wattled honeyeater. Sporulated oocysts are ovoid, 28.9 × 26.1 (25-32 × 23-30) µm, with a smooth, colorless, bilayered wall; the inner wall is slightly thicker and darker than the outer wall. The average shape index is 1.1. No micropyle or oocyst residuum are present but the oocyst contains one or two ovoid polar granules. Sporocysts are ovoid, 17.1 × 10.9 (16-18 × 10-11) µm with a smooth single layered wall and an average shape index of 1.6. The Stieda body is broad, dome-like with a rather rectangular-shaped substieda body. Within the sporocyst is a large amorphous residuum composed of coarse granules and 4 randomly arranged, sausage-shaped sporozoites with a subspherical, posterior refractile body and a centrally located nucleus.

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

    Science.gov (United States)

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

    2007-11-15

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

  3. Crab-eating fox (Cerdocyon thous), a South American canid, as a definitive host for Hammondia heydorni.

    Science.gov (United States)

    Soares, Rodrigo M; Cortez, Luiz R P B; Gennari, Solange M; Sercundes, Michelle K; Keid, Lara B; Pena, Hilda F J

    2009-05-26

    Hammondia heydorni is a cyst forming coccidia closely related to other apicomplexans, such as Toxoplasma gondii, Neospora caninum and Hammondia hammondi with a two-host life cycle. Dogs and other canids as red foxes (Vulpes vulpes) and coyotes (Canis latrans) may serve as definitive hosts for H. heydorni. Sporulated oocysts are infective for cattle, sheep and goats, which may serve as intermediate hosts. Herein, we describe the ability of crab-eating fox (Cerdocyon thous), a wild carnivore that is commonly found from northern Argentina to northern South America, to serve as definitive host of H. heydorni. The whole masseter muscle and brain from two 2-year-old bovines were collected, minced and pooled together for the fox infection. The bovine pooled tissues were equally administered to four foxes, in two consecutive days. Two foxes shed subspherical unsporulated oocysts measuring 10-15microm, after 8 and 9 days post-infection, respectively. One of the foxes eliminated oocysts for 5 days, while the other fox shed oocysts for 9 days. A DNA sample of oocysts detected at each day of oocyst elimination was tested by two PCRs, one of them carried out employing primers directed to the common toxoplasmatiid 18S and 5.8S ribosomal RNA coding genes (PCR-ITS1) and the other based on heat-shock protein 70kDa coding gene (PCR-HSP70). These samples were also submitted to a N. caninum specific nested-PCR protocol based on a N. caninum specific gene (Nc5-nPCR). All of them were positive by PCR-ITS1 and PCR-HSP70 but negative by Nc5-nPCR. The PCR-ITS1 and PCR-HSP70 nucleotide sequences amplified from the oocysts shed by the foxes revealed 100% identity with homologous sequences of H. heydorni. In conclusion, it is clear that H. heydorni also uses the crab-eating fox as a definitive host. The crab-eating fox is usually reported to live in close contact with livestock in several regions of Brazil. Therefore, it is reasonable to infer that such carnivores may play an important role

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

    Science.gov (United States)

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

    2002-08-01

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

  5. Outcome of an artificial coccidial infection in poults under the influence of floor heating.

    Science.gov (United States)

    Abd El-Wahab, A; Visscher, C F; Wolken, S; Reperant, L-M; Beineke, A; Beyerbach, M; Kamphues, J

    2013-03-01

    Coccidiosis is one of the most prevalent diseases in poultry. The objective of the current study was to determine the effects of floor heating when poults were artificially infected with Eimeria regarding the outcome of the infection and secondary effects on litter quality and health of the foot pad. Two trials were performed. In each trial, 4 groups of 2-wk-old turkeys were reared for a 4-wk period. All birds were fed ad libitum identical pelleted diets without any anticoccidial additive. The first 2 groups were housed on dry wood shavings, with and without floor heating; the other 2 groups were housed on wet wood shavings (35% moisture, achieved by adding water as required), with and without floor heating. Two birds only (primary seeder birds; the other 18 birds were nominated as secondary infected birds) in each of the 4 groups were experimentally infected orally with Eimeria adenoeides (~50,000 oocysts/bird). The number of oocysts eliminated via excreta was determined repeatedly. On d 42, each bird not only was scored macroscopically for coccidial lesions in the cecum but also oocyst counts were determined in the cecal contents. Finally, the foot pads were assessed weekly for external scoring and on d 42 of life for histopathological scoring. Although the counts of oocysts in seeder birds were almost identical (d 6 postinoculation), oocyst counts in excreta of secondary birds were markedly reduced in both trials when the poults were not exposed to wet litter. Moreover, in both trials using floor heating with exposure to wet litter resulted in a higher oocyst count in the excreta of secondary infected birds (3.72/3.92 in trials 1 and 2) on d 24 postinoculation compared with the other groups. Using floor heating resulted in significantly decreased foot pad dermatitis scores compared with groups housed without floor heating. In conclusion, the differences in oocyst counts indicate that the process of sporulation is affected by both the moisture and the

  6. Isospora streperae n. sp. (Apicomplexa: Eimeriidae) from a grey currawong (Strepera versicolour plumbea) (Passeriformes: Artamidae) in Western Australia.

    Science.gov (United States)

    Yang, Rongchang; Brice, Belinda; Habsi, Khalid Al; Elliot, Aileen; Ryan, Una

    2015-01-01

    A new species, Isospora streperae n. sp., (Apicomplexa: Eimeriidae) is described from a single grey currawong bird (Strepera versicolour) (subspecies S. v. plumbea) in Western Australia. Sporulated oocysts (n = 32) are spherical to subspherical, with smooth colourless bilayered oocyst wall, 1.0 µm thick (outer layer 0⋅8 µm, inner 0.2 µm thick). Oocyst with a polar granule, an oocyst residuum and two spheroidal to subspheroidal sporocysts. Oocyst length, 23.8 (20.4-25.0) µm; oocyst width, 22.5 (20.0-24.6) µm; a shape index of 1.06, with Stieda, substieda bodies. Micropyle is absent. Sporocysts with compressed sporocyst residuum and four sporozoites. Sporocyst length, 14.4 (12.5-15.2) µm; sporocyst width, 11.2 (10.6-14.0) µm, sporocyst L/W ratio, 1.29. Necropsy of the bird identified haemorrhaging along the ileum and jejunum, which is where Isospora oocysts were also mostly detected. Molecular analysis was conducted at three loci; the 18S, 28S ribosomal RNA and the mitochondrial cytochrome oxidase (COI) gene. At the 18S locus, I. streperae n. sp. exhibited 99.5% and 99.4% similarity respectively to an Isospora sp. (MS-2003) from a Southern cape sparrow (Passer melanurus melanurus) and Isospora dovati from a domestic pigeon (Columba livia domestica). At the 28S locus, I. streperae n. sp. exhibited 96.9% similarity to an Isospora sp. (MS-2003) from a grosbeak starling (Scissirostrum dubium) and 95.8% similarity with the Isospora sp. (MS-2003) from a Southern cape sparrow. At the COI locus, I. streperae n. sp. exhibited 95.0% similarity to Isospora sp. from a yellow-necked mouse (Apodemus flavicollis) from the Czech Republic. Based on morphological and molecular data, this isolate is a new species of Isospora, which is named Isospora streperae n. sp. after its host, the grey currawong (Strepera versicolour plumbea). Crown Copyright © 2014. Published by Elsevier Inc. All rights reserved.

  7. Synergistic approach for treatment of chicken coccidiosis using berberine--A plant natural product.

    Science.gov (United States)

    Malik, Tauseef Ahmad; Kamili, Azra N; Chishti, M Z; Tanveer, Syed; Ahad, Shazia; Johri, R K

    2016-04-01

    Despite the advent of anticoccidial drugs and vaccines, coccidiosis continues to result in substantial economic losses to the poultry industry. Berberine, a natural alkaloid is well known in studies involving synergistic approaches, thereby reducing the dosage of principal drugs. Therefore, a study was designed to see whether a synergistic anticoccidial effect could be obtained between amprolium and berberine, in vivo using broiler chicken. Anticoccidial activity was measured in comparison to the reference drug amprolium on the basis of oocyst output reduction, mean weight gain and feed conversion ratio. Oocyst output was measured using Mc-Masters counting technique. Different combinations of berberine and amprolium were tested and out of which 1:1 ratio was the most effective for controlling these parasites. Oral gavaging of 100(50 + 50) mg/kg body weight of 1:1 ratio of amprolium and berberine caused the equivalent reduction in number of oocysts (38.85 ± 9.61) one day prior to that of standard drug amprolium (49.95 ± 16.65) as well as pure berberine (44.4 ± 9.61) used in the study. Weight gain of birds was also highest in the synergistic group (1547.43 ± 12.86) among all the infected groups. Besides feed conversion ratio in the synergistic group was also better (1.387 ± 0.026). The results of this study proved the effectiveness of both amprolium and berberine and revealed synergism between amprolium and berberine against coccidian oocysts, confirmed by significant reduction in the number of coccidian oocysts shed in the feces, leading to better weight gain and improved feed conversion ratio. The study deep-rooted the synergistic potential of berberine, a natural bioactive compound for controlling a protozoan parasite and the results of this study corroborate with its use for treatment of severe diarrhoea, amoebiasis and intestinal infections.

  8. Amphiphile disruption of pathogen attachment at the hematite (α-Fe2O3)-water interface.

    Science.gov (United States)

    Gao, Xiaodong; Chorover, Jon

    2011-05-17

    Prior studies have indicated that the subsurface transport of Cryptosporidium parvum oocysts is diminished in sediments containing iron oxides and that inner-sphere complexation of oocyst surficial carboxylate plays a role in the retardation. However, the impacts of natural organic matter (NOM) remain poorly understood. In this study, we used a model anionic surfactant, sodium dodecyl sulfate (SDS), as a surrogate for amphiphilic NOM components to examine the impacts of amphiphilic components on oocyst adhesion mechanisms. We employed in situ attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy to determine the effects of SDS on the molecular bonds that mediate interactions between oocyst surficial biomolecules and hematite (α-Fe(2)O(3)) surface functional groups over a wide range of solution pH. The results show that the presence of SDS significantly diminishes Fe-carboxylate complexation, as indicated by progressive decreases in intensity of asymmetric and symmetric stretching vibrations of carboxylate [ν(as)(COO(-)) and ν(s)(COO(-))] with reaction time. In addition, one of the ν(s)(COO(-)) bands shifted from 1370 to 1418 cm(-1) upon SDS introduction, suggesting that SDS also changed the complexation mode. The data indicate that competition from the sulfonate groups (OSO(3)(-)) of SDS at α-Fe(2)O(3) surface sites is a primary mechanism resulting in decreased Fe-carboxylate complexation. Sorptive competition from amphiphilic NOM components may therefore increase the mobility of C. parvum oocysts in the environment through disruption of interfacial pathogen-mineral surface bonds.

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

    Science.gov (United States)

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

    2016-06-01

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

  10. Large-scale survey of the prevalence of Eimeria infections in domestic rabbits in China.

    Science.gov (United States)

    Jing, Fa; Yin, Guangwen; Liu, Xianyong; Suo, Xun; Qin, Yinghe

    2012-04-01

    The current study examined the prevalence of Eimeria infections in domestic rabbits in China. A total of 480 faecal samples were collected from 48 farms in 14 provinces of China. Each faecal sample was subjected to oocyst counting and oocyst isolation. The Eimeria species from samples containing isolated and sporulated oocysts were morphologically identified under microscope. The overall prevalence of infections was 41.9% (201/480). Northwest China had the highest prevalence (70%), followed closely by Northeast China (65%) and Southwest China (62.5%). The prevalences in North China (34%) and South China (25.8%) were significantly lower. The large and medium farms had lower prevalences (34.2% and 37.2%, respectively) than the small farms (61.4%). Coccidian oocysts were found in 42.2% (76/180) of faecal samples from meat rabbits, 40% (28/70) from angora rabbits and 44.7% (85/190) from Rex rabbit. In total, ten species of Eimeria were identified from oocyst-positive samples. Concurrent infection with two to eight Eimeria species was found. E. perforans was the most prevalent species (35.2%), followed in order by E. media, E. magna, E. irresidua and E. intestinalis with prevalences of 31.3%, 28.8%, 19.4%, and 14.8%, respectively. Taken together, These results reveal the characteristics of the prevelance of rabbit coccidia infection in China, including the distribution, the scale of farming and the species, which are indispensable to the control of rabbits coccidiosis in China.

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

    Science.gov (United States)

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

    2013-09-01

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

  12. Large-scale investigation of the parameters in response to Eimeria maxima challenge in broilers.

    Science.gov (United States)

    Hamzic, E; Bed'Hom, B; Juin, H; Hawken, R; Abrahamsen, M S; Elsen, J M; Servin, B; Pinard-van der Laan, M H; Demeure, O

    2015-04-01

    Coccidiosis, a parasitic disease of the intestinal tract caused by members of the genera Eimeria and Isospora, is one of the most common and costly diseases in chicken. The aims of this study were to assess the effect of the challenge and level of variability of measured parameters in chickens during the challenge with Eimeria maxima. Furthermore, this study aimed to investigate which parameters are the most relevant indicators of the health status. Finally, the study also aimed to estimate accuracy of prediction for traits that cannot be measured on large scale (such as intestinal lesion score and fecal oocyst count) using parameters that can easily be measured on all animals. The study was performed in 2 parts: a pilot challenge on 240 animals followed by a large-scale challenge on 2,024 animals. In both experiments, animals were challenged with 50,000 Eimeria maxima oocysts at 16 d of age. In the pilot challenge, all animals were measured for BW gain, plasma coloration, hematocrit, and rectal temperature and, in addition, a subset of 48 animals was measured for oocyst count and the intestinal lesion score. All animals from the second challenge were measured for BW gain, plasma coloration, and hematocrit whereas a subset of 184 animals was measured for intestinal lesion score, fecal oocyst count, blood parameters, and plasma protein content and composition. Most of the parameters measured were significantly affected by the challenge. Lesion scores for duodenum and jejunum (P Eimeria maxima. Prediction of intestinal lesion score and fecal oocyst count using the other parameters measured was not very precise (R2 Eimeria maxima has a strong genetic determinism, which may be improved by genetic selection.

  13. Assessment of humoral immunity to Eimeria tenella sporozoites in chickens by ELISA

    Directory of Open Access Journals (Sweden)

    S. Saravanan

    2014-07-01

    Full Text Available Aim: To assess the humoral immune response of Eimeria tenella sporozoites in broiler chickens by a developed enzyme linked immunosorbent assay (ELISA and the efficacy in terms of bodyweight, lesion score and oocysts excretion in immunized broilers. Materials and Methods: Purified live E. tenella sporozoites were administered subcutaneously in neck region of broiler chickens in the early life (first week at different concentrations. The potency of the sporozoite vaccine as assessed by IgG levels and the performance in immunized broilers as assessed by body weight, lesion score and oocysts excretion in faeces after challenge with 10, 000 live E. tenella oocysts at 49 days of age were evaluated. Results: The chickens of group (T4 immunized with 20 µg of antigen on day 6 showed an increase in IgG levels (0.161±0.004 two weeks post immunization (PI peaking (0.399± 0.016 at 5 weeks PI. The mean weekly weight gain (g after challenge, at 56 days of age was high in T4 (148±4.751 g with a low mean lesion score (2.5±0.22 and mean oocyst output (x103 oocytes per gram (OPG in faeces (100.3± 45.72 when compared to unimmunised infected controls. Conclusion: An early but partial immune response against caecal coccidiosis could be achieved by immunization with E. tenella specific sporozoites in chickens of less than a week old. Moreover, the performance of immunized chickens as indicated by weight gain, lesion score and oocyst output was found to be superior to the unimmunized infected controls.

  14. Sequence-based genotyping clarifies conflicting historical morphometric and biological data for 5 Eimeria species infecting turkeys.

    Science.gov (United States)

    El-Sherry, S; Ogedengbe, M E; Hafeez, M A; Sayf-Al-Din, M; Gad, N; Barta, J R

    2015-02-01

    Unlike with Eimeria species infecting chickens, specific identification and nomenclature of Eimeria species infecting turkeys is complicated, and in the absence of molecular data, imprecise. In an attempt to reconcile contradictory data reported on oocyst morphometrics and biological descriptions of various Eimeria species infecting turkey, we established single oocyst derived lines of 5 important Eimeria species infecting turkeys, Eimeria meleagrimitis (USMN08-01 strain), Eimeria adenoeides (Guelph strain), Eimeria gallopavonis (Weybridge strain), Eimeria meleagridis (USAR97-01 strain), and Eimeria dispersa (Briston strain). Short portions (514 bp) of mitochondrial cytochrome c oxidase subunit I gene (mt COI) from each were amplified and sequenced. Comparison of these sequences showed sufficient species-specific sequence variation to recommend these short mt COI sequences as species-specific markers. Uniformity of oocyst features (dimensions and oocyst structure) of each pure line was observed. Additional morphological features of the oocysts of these species are described as useful for the microscopic differentiation of these Eimeria species. Combined molecular and morphometric data on these single species lines compared with the original species descriptions and more recent data have helped to clarify some confusing, and sometimes conflicting, features associated with these Eimeria spp. For example, these new data suggest that the KCH and KR strains of E. adenoeides reported previously represent 2 distinct species, E. adenoeides and E. meleagridis, respectively. Likewise, analysis of the Weybridge strain of E. adenoeides, which has long been used as a reference strain in various studies conducted on the pathogenicity of E. adenoeides, indicates that this coccidium is actually a strain of E. gallopavonis. We highly recommend mt COI sequence-based genotyping be incorporated into all studies using Eimeria spp. of turkeys to confirm species identifications and so

  15. A comparative transcriptome analysis reveals expression profiles conserved across three Eimeria spp. of domestic fowl and associated with multiple developmental stages.

    Science.gov (United States)

    Novaes, Jeniffer; Rangel, Luiz Thibério L D; Ferro, Milene; Abe, Ricardo Y; Manha, Alessandra P S; de Mello, Joana C M; Varuzza, Leonardo; Durham, Alan M; Madeira, Alda Maria B N; Gruber, Arthur

    2012-01-01

    Coccidiosis of the domestic fowl is a worldwide disease caused by seven species of protozoan parasites of the genus Eimeria. The genome of the model species, Eimeria tenella, presents a complexity of 55-60MB distributed in 14 chromosomes. Relatively few studies have been undertaken to unravel the complexity of the transcriptome of Eimeria parasites. We report here the generation of more than 45,000 open reading frame expressed sequence tag (ORESTES) cDNA reads of E. tenella, Eimeria maxima and Eimeria acervulina, covering several developmental stages: unsporulated oocysts, sporoblastic oocysts, sporulated oocysts, sporozoites and second generation merozoites. All reads were assembled to constitute gene indices and submitted to a comprehensive functional annotation pipeline. In the case of E. tenella, we also incorporated publicly available ESTs to generate an integrated body of information. Orthology analyses have identified genes conserved across different apicomplexan parasites, as well as genes restricted to the genus Eimeria. Digital expression profiles obtained from ORESTES/EST countings, submitted to clustering analyses, revealed a high conservation pattern across the three Eimeria spp. Distance trees showed that unsporulated and sporoblastic oocysts constitute a distinct clade in all species, with sporulated oocysts forming a more external branch. This latter stage also shows a close relationship with sporozoites, whereas first and second generation merozoites are more closely related to each other than to sporozoites. The profiles were unambiguously associated with the distinct developmental stages and strongly correlated with the order of the stages in the parasite life cycle. Finally, we present The Eimeria Transcript Database (http://www.coccidia.icb.usp.br/eimeriatdb), a website that provides open access to all sequencing data, annotation and comparative analysis. We expect this repository to represent a useful resource to the Eimeria scientific

  16. The Alice - "Follow the White Rabbit" - parasites of farm rabbits based on coproscopy.

    Science.gov (United States)

    Kornaś, Sławomir; Kowal, Jerzy; Wierzbowska, Izabela; Basiaga, Marta; Nosal, Paweł; Niedbała, Piotr

    2015-01-01

    The aim of the study, conducted in the years 2011-2013, was to determine the level of gastrointestinal parasites infection in New Zealand White rabbits, kept at the Experimental Station of the University of Agriculture in Krakow. The study showed rabbits protozoan infection with the genus Eimeria, belonging - based on the sporulation method - to the following species: E. magna, E. media, E. perforans, E. stiedae and E. irresidua. The highest prevalence of infection, as well as the intensity of oocysts output (OPG - oocysts per gram of faeces), was noted for E. magna and E. media - respectively 31.4 % (19477.3 OPG), and 40.0 % (14256.07 OPG). The infection of rabbits with Eimeria spp. differed significantly between years. With regard to oocysts output, the level of infection was strongly connected with the age of rabbits, being higher in young animals. However, the range of infection was highest among adults. Among nematodes, Passalurus ambiguus pinworm was regularly found (prevalence reached 21.9%), other species - Trichuris leporis, and Graphidium strigosum were rarely noted. The overall infection with nematodes did not differ between years. Similarly, as in the case of Eimeria older individuals were more often infected by nematodes. We observed some trends in parasite oocysts/eggs output; the protozoan oocysts were recorded more often in faecal samples collected in the evenings, whereas the nematodes eggs occurred frequently in the mornings. This situation may be related to the phenomenon of coprophagy occurring in the mammals of Lagomorpha order. The results of the study indicate that especially coccidiosis constitute permanently throughout the years an important problem in the rabbitry examined.

  17. Efficacy of emodepside/toltrazuril suspension (Procox® oral suspension for dogs) against mixed experimental Isospora felis/Isospora rivolta infection in cats.

    Science.gov (United States)

    Petry, Gabriele; Kruedewagen, Eva; Kampkoetter, Andreas; Krieger, Klemens

    2011-08-01

    The coccidia Isospora felis and Isospora rivolta are intestinal parasites occurring worldwide in domestic cats. In young cats, they can be detected with higher prevalence.The effects of toltrazuril in the new combination product Procox(®) oral suspension for dogs containing 0.1 % emodepside and 2 % toltrazuril (0.9 mg emodepside + 18 mg toltrazuril per ml) were studied in eighteen kittens experimentally infected each with a total of 1 x 10(5) oocysts of a mixture of Isospora felis and Isospora rivolta. In the infectious material, the quantitative relation of I. felis and I. rivolta was about 1:5. Following a three-days period after infection, two groups of 6 kittens were treated during the prepatent period with either a single dose of 0.45 mg emodepside + 9 mg toltrazuril/kg body weight or 0.9 mg emodepside + 18 mg toltrazuril/kg body weight. A group of six kittens without any treatment served as a control. On day 5 post infection, the untreated kittens started the excretion of oocysts. Treatment with both toltrazuril doses significantly reduced oocyst excretion. Following the single higher dose, the reduction of oocysts of both Isospora spp. was more pronounced (96.7 % to 100 %) in comparison to the lower dose (57.2 % to 100 %). The Procox(®) application was well tolerated and no adverse events were seen with any of the applied dosages.When administered to kittens and as a single treatment during the prepatent period, Procox(®) is suitable to control the number of oocysts excreted in the faeces in case of an Isospora felis and Isospora rivolta infection.

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

    Science.gov (United States)

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

    2012-12-01

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

  19. Use of ponazuril paste to treat coccidiosis in shelter-housed cats and dogs.

    Science.gov (United States)

    Litster, A L; Nichols, J; Hall, K; Camp, J; Mohamed, A S

    2014-05-28

    Cystoisospora (synonym Isospora) spp. infections are common in dogs and cats worldwide, especially in crowded or unsanitary environments. Ponazuril (toltrazuril sulfone) is a widely used oral treatment, but protocols that will produce oocyst excretion below the detection limit in shelter-housed animals have not been determined. The aim of this study was to determine the efficacy of ponazuril paste at each of three dosages (dosage 1, 50mg/kg q24 h for 3 days, dogs n=14, cats n=16; dosage 2, 50mg/kg as a single dose, dogs n=13, cats n=25; or dosage 3, 20mg/kg as a single dose, dogs n=16, cats n=23) in shelter-housed dogs (n=43) and cats (n=64) with confirmed coccidiosis. Fecal oocyst counts and identification and fecal consistency scoring was performed pre-treatment (Day 1) and again at Day 3-4 and Day 8. There were higher proportions of animals with oocyst excretion below the detection limit at both Day 3-4 and Day 8 in the dosage 1 group (dogs 92.9%, cats 87.5%) than in the other two groups (dosage 2, dogs 76.9%, cats 80.0%; dosage 3, dogs 68.8%, cats 47.8%). Animals with high fecal oocyst counts at Day 1 were significantly more likely to be infected at Day 3-4 (dogs, Pcats, P=0.013). Fecal consistency score at Day 3-4 was not significantly related to infection status (dogs, P=0.898; cats, P=0.136). Further studies are warranted to investigate a ponazuril protocol that can safely reduce fecal oocyst burdens in infected dogs and cats to levels below the detection limit. Environmental decontamination is also important to reduce the likelihood of re-infection. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. Morphological and molecular characterization of Choleoeimeria pogonae n. sp. coccidian parasite (Apicomplexa: Eimeriidae, 1989, Paperna and Landsberg) in a western bearded dragon (Pogona minor minor).

    Science.gov (United States)

    Yang, Rongchang; Brice, Belinda; Ryan, Una

    2016-01-01

    A new species, Choleoeimeria pogonae n. sp. is described from a Western bearded dragon (Pogona minor minor) in Western Australia. Sporulated oocysts (n = 48) were cylindroidal in shape. Oocyst length, 27.0 (26.0-28.3) μm, oocyst width, 15.2 (14.0-16.5) μm, oocyst length/width ratio (L/W) 1.8 (1.6-1.9), each with 4 sporocysts (Eimeria-like) and a polar granule, but lacking a micropyle and oocyst residuum. Sporocysts are ovoidal in shape, sporocyst length, 10.0 (9.0-11.0) μm, sporocyst width 8.5 (7.0-9.5) μm, sporocyst L/W ratio, 1.2 (1.1-1.3). Stieda, substieda and parasubstieda bodies were all absent. Molecular analysis was conducted at the 18S rRNA and cytochrome c oxidase I (COI) loci. Phylogenetic analysis of 18S sequences revealed that C. pogonae n. sp. grouped together with another four Choleoeimeria spp. and exhibited 99.1%-99.4% genetic similarity. At the COI locus, C. pogonae n. sp. was in an independent clade and had the highest similarity (80.4%) to Eimeria cf. mivati from a chicken (Gallus gallus domesticus). According to the morphological and molecular data, this isolate is a new species of coccidian parasite. This study further supports the taxonomy of Choleoeimeria spp. as a new genus based on molecular phylogenetic analysis. Crown Copyright © 2015. Published by Elsevier Inc. All rights reserved.

  1. Six new species of coccidia (Apicomplexa: Eimeriidae) from endangered Phelsuma spp. geckoes (Sauria: Gekkonidae) of the Black River Gorges National Park, Mauritius.

    Science.gov (United States)

    Daszak, Peter; Ball, Stanley J; Jones, Carl G; Streicker, Daniel G; Snow, Keith R

    2009-12-01

    Six new species of coccidia are described from endangered Phelsuma spp. geckoes (Sauria: Gekkonidae) endemic to Mauritius, Indian Ocean. Five new species (3 Eimeria and 2 Isospora species) are described from Phelsuma rosagularis Vinson et Vinson; all lack a micropyle and an oocyst residuum, and all have a sporocyst residuum. Oocysts of Eimeria swinnertonae sp. n. are ellipsoidal, 22.2 x 17.8 (20.8-24.8 x 16.8-18.4) microm; SI 1.25; polar granule absent. Sporocysts are ellipsoidal, 8.8 x 7.0 (8.0-9.6 x 6.4-8.0) microm; SI 1.3; Stieda body absent. Oocysts of Eimeria stebbinsi sp. n. are ellipsoidal, 17.4 x 11.7 (16.0-19.2 x 11.2-12.8) microm; SI 1.5; polar granules present. Sporocysts are elongate-ellipsoidal, 7.7 x 4.0 (7.2-8.0 x 3.2-5.6) microm; SI 1.9; Stieda body present. Oocysts of Eimeria raleighi sp. n. are spheroidal to sub-spheroidal, 17.0 x 15.5 (16.0-19.2 x 14.4-16.8) microm; SI 1.1; polar granule present. Sporocysts are sub-spheroidal, 7.8 x 6.6 (7.2-8.0 x 6.4-7.2) microm; SI 1.2; Stieda body absent. Oocysts of Isospora cottinghamae sp. n. are ellipsoidal, 19.8 x 15.5 (17.6-21.6 x 14.4-17.6) microm; SI 1.3; polar granules present. Sporocysts are ellipsoidal, 10.8 x 6.9 (9.6-12.8 x 6.4-8.0) microm; SI 1.6; Stieda body present. Oocysts of Isosporapearlae sp. n. are ellipsoidal, 16.0 x 11.5 (15.2-17.6 x 9.6-12.8) microm; SI 1.4; polar granule present. Sporocysts are ellipsoidal, 8.8 x 5.4 (8.0-9.6 x 4.8-6.4) microm; SI 1.6; Stieda and substieda bodies present. One new Eimeria species is described from the blue-tailed day gecko, Phelsuma cepediana Merrem. Oocysts of Eimeria hartleyi sp. n. are sub-spheroidal to ellipsoidal, 18.2 x 14.5 (16.0-20.8 x 13.6-16.0) microm; SI 1.26; polar granules present. Sporocysts are ellipsoidal to cylindroidal, 7.5 x 5.3 (6.4-8.0 x 4.8-6.4) microm; SI 1.4; Stieda body present. We report the presence of tetrazoic spheroidal to sub-spheroidal oocysts or sporocysts 10.2 x 8.5 (9.9-10.4 x 8.3-8.8) microm; SI 1.2 from an individual

  2. Evaluation of an Improved U.S. Food and Drug Administration Method for the Detection of Cyclospora cayetanensis in Produce Using Real-Time PCR.

    Science.gov (United States)

    Murphy, Helen R; Lee, Seulgi; da Silva, Alexandre J

    2017-07-01

    Cyclospora cayetanensis is a protozoan parasite that causes human diarrheal disease associated with the consumption of fresh produce or water contaminated with C. cayetanensis oocysts. In the United States, foodborne outbreaks of cyclosporiasis have been linked to various types of imported fresh produce, including cilantro and raspberries. An improved method was developed for identification of C. cayetanensis in produce at the U.S. Food and Drug Administration. The method relies on a 0.1% Alconox produce wash solution for efficient recovery of oocysts, a commercial kit for DNA template preparation, and an optimized TaqMan real-time PCR assay with an internal amplification control for molecular detection of the parasite. A single laboratory validation study was performed to assess the method's performance and compare the optimized TaqMan real-time PCR assay and a reference nested PCR assay by examining 128 samples. The samples consisted of 25 g of cilantro or 50 g of raspberries seeded with 0, 5, 10, or 200 C. cayetanensis oocysts. Detection rates for cilantro seeded with 5 and 10 oocysts were 50.0 and 87.5%, respectively, with the real-time PCR assay and 43.7 and 94.8%, respectively, with the nested PCR assay. Detection rates for raspberries seeded with 5 and 10 oocysts were 25.0 and 75.0%, respectively, with the real-time PCR assay and 18.8 and 68.8%, respectively, with the nested PCR assay. All unseeded samples were negative, and all samples seeded with 200 oocysts were positive. Detection rates using the two PCR methods were statistically similar, but the real-time PCR assay is less laborious and less prone to amplicon contamination and allows monitoring of amplification and analysis of results, making it more attractive to diagnostic testing laboratories. The improved sample preparation steps and the TaqMan real-time PCR assay provide a robust, streamlined, and rapid analytical procedure for surveillance, outbreak response, and regulatory testing of foods for

  3. Isolation and identification of Eimeria tenella Anhui Feixi strain%柔嫩艾美耳球虫安徽肥西(AHFX)株的分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    张龙乐; 徐前明; 李培英

    2012-01-01

    目的 分离并鉴定安徽肥西病鸡盲肠内的柔嫩艾美耳球虫.方法 通过雏鸡进行单卵囊接种与增殖试验,对所获卵囊用形态学方法进行初步鉴定;运用PCR方法扩增该分离株的ITS-1基因序列,并与相关序列进行比对、构建系统进化树.结果 该球虫分离株孢子化卵囊的平均大小为21.1 μm ×18.0 μm,卵形指数为1.17,潜隐期为140 h;其ITS-1基因序列与柔嫩艾美耳球虫(Eimeria tenella)GQ856310相似性达98.2%,二者的亲缘关系非常接近.结论 该球虫分离株为柔嫩艾美耳球虫(E.tenella),暂命名为柔嫩艾美耳球虫安徽肥西(AHFX)株.%Objective To isolate and identify the Eimeria tenella from Eimeria oocysts of chickens obtained from Feixi County, Anhui Province. Methods The pure strain was obtained with single oocyst method and proliferated. After identifying the oocysts by using morphological methods, the internal transcribed spacer 1 (ITS-1) was amplified by polymerase chain reaction (PCR) and sequenced, then the phylogenetic tree analysis of the sequence was performed to determine the species. Results The average size of the sporulated oocysts was 21.1 μm × 18.0 μm; the oocyst index was 1.17, and the latent period was 140 h. The homology in ITS-1 was 98. 2% between the isolated strain and E. tenella (GQ856310); they were very close in phylogeny. Conclusion The isolated oocyst was identified as E. tenella Anhui Feixi strain.

  4. Systemic acute phase proteins response in calves experimentally infected with Eimeria zuernii.

    Science.gov (United States)

    Lassen, Brian; Bangoura, Berit; Lepik, Triin; Orro, Toomas

    2015-09-15

    Acute phase proteins (APPs) have been demonstrated to be useful in evaluating general health stress and diseases in cattle. Serum amyloid A (SAA) and haptoglobin (Hp) are APPs that are produced during inflammation, and likely play a role in host immunological defence against Eimeria infection and the associated intestinal tissue damage. We investigated the involvement of SAA and HP in an experimental study, including three groups of calves: a control group (group 0, n=11), and two groups infected with either 150,000 or 250,000 Eimeria zuernii oocysts (group 1 (n=11) and group 2 (n=12), respectively). The calves were monitored for 28 days and data was collected on oocyst excretion, faecal score, animal weight, and SAA and Hp serum concentrations. Generalized linear mixed models showed that the clinical symptoms, indicated by an increase in the number of oocysts in the faeces and severe diarrhoea, manifested at patency for group 1 and 2. Serum Hp and SAA levels also increased during this period. Hp appeared to be a more sensitive marker than SAA, and differences between groups 1 and 2 were observed only for Hp. Linear regression models showed a negative association between weight gain and Hp concentrations, calculated as the area under the curve (AUC) during the overall experimental period and the patency period. A similar result was seen for SAA only during the patency period. This result supports the assumption that reduced weight gain due to E. zuernii infection is an immunologically driven process that involves an increase in APPs. A random intercept regression model of oocyst shedding groups showed that calves shedding 1-500 oocysts had reduced concentrations of Hp, indicating that a different immunological reaction occurs during mild shedding of E. zuernii oocysts than during more intensive shedding. A similar model was used to examine associations between faecal scores and Hp concentrations for each group. Group 2 calves with haemorrhagic diarrhoea displayed

  5. Epidemiological survey of cryptosporidiosis in Anhui Province China

    Institute of Scientific and Technical Information of China (English)

    Ke-Xia Wang; Chao-Pin Li; Jian Wang; Bo-Rong Pan

    2002-01-01

    AIM: To provide scientific evidence for prevention and controlling of cryptosporidiosis, the infection of Cryptosporidium parvum and its epidemiological characteristics were studied in some areas of Anhui Province.METHODS: The oocyst of Cryptosporidium parvum in 5421fresh stool samples from eleven areas of Anhui Provincewas tested by auramine-phenol stain and improved anti-acidstain respectively. The specific antibody of IgG, IgM and Tsubsets of 41 patients with positive Cryptosporidium parvumin stools were detected by ELISA and biotin-streptavidin(BSA) respectively.RESULTS: The total infective rate of Cryptosporidiumparvum was 1.33 % (74/5421). Among them, the positiverates of oocyst in the areas of Huaibei (1. 82 % ) and Fuyang( 1. 80 % ) were higher. The positive rates of oocyst in stoolsof infants, pupils, middle school students, collegestudents, adults, patients with diarrhea, and those withimmunodeficiency were 3.15 % (28/889), 0.82 % (9/1098), 0.82 %(9/1092), 0.83 %(8/969), 0.85 % (9/1095), 2.88 %(8/27g) and 8.33 %(3/36) % respectively. The positive rates ofoocyst in infants and the patients with diarrhea andimmunodeficiency were significantly higher than those incontrols ( P < 0.01 ). The positive rate of oocyst in maleswas similar to that in females ( P> 0.05). The positive rateof oocyst in urban areas ( 1. 13 %) was significantly lowerthan those in rural areas ( 1. 72 %, P < 0.01 ). The positiverates of specific IgG, IgM and IgG + IgM in sera of thepatients with positive oocyst in stool were 63.4 % ( 26/41 ),17.1% (7/41), 19.5 % (8/41) respectively. The numberfractions of T subsets of CD3 + , CD4 + , CD8 + and CD4 +/CD8 + of the patients were 0.66 ± 0.07, 0.44 ± 0.06, 0.28 ± 0.04 and 1. 58 ± 0.32 respectively. The difference between thepatients and the controls was significant ( P < 0. 05). Themain manifestations of the patients were subclinicalinfection, in forms of slight abdominal pain, mild diarrhea,and loose stool.CONCLUSION: There are

  6. Coccidiosis in japanese quails (Coturnix japonica: characterization of a naturally occurring infection in a commercial rearing farm

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    M Teixeira

    2004-06-01

    Full Text Available A study about coccidiosis in Japanese quails was carried out in order to identify species of the genus Eimeria and characterize a naturally occurring infection in a commercial rearing farm. For this purpose, fecal exams, oocyst counting and morphological study were performed, besides necropsy and histopathology to confirm diagnosis. Three species of the genus Eimeria were found and identified as E. tsunodai, E. uzura and E. bateri. The natural infection was characterized as subclinical because of the mild and nonspecific clinical signs. Nevertheless, coccidiosis was considered an important disease because endogenous stages of the parasites and a high number of oocysts in feces were associated with intestinal lesions. The results suggest that such infection might represent a limiting factor to this branch of the modern poultry industry.

  7. High occurrence of cyclosporiasis in Istanbul, Turkey, during a dry and warm summer

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    Hakko Elif

    2010-04-01

    Full Text Available Abstract We evaluated the incidence of Cyclospora cayetanensis in immunocompetent, diarrheic patients during the summers of 2006-2009 in Istanbul. Stools from 1876 patients were examined using microscopic techniques. Cyclospora oocysts were observed in wet preparations by light and epifluorescence microscopy and in fecal smears that were stained by Kinyoun's modified acid-fast stain. Characteristic Cyclospora oocysts were observed in 2 patients in 2006, 17 in 2007, and one in 2009. Samples positive for Cyclospora were further analyzed by a single step polymerase chain reaction (PCR with Cyclospora-specific primers from the ITS-1 region of the genome. The majority of the Cyclospora positive cases (15 were clustered during about 15 days in June 2007, indicating an unusual incidence of cyclosporiasis in this time period. The climatic characteristics of 2007 could have played a role in this high occurrence rate.

  8. Four new species of Eimeria (Apicomplexa: Eimeriidae) from Emoia spp. Skinks (Sauria: Scincidae), from Papua New Guinea and the Insular Pacific

    Science.gov (United States)

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

    2017-01-01

    Between September and November 1991, 54 adult skinks from 15 species were collected by hand or blowpipe from several localities on Rarotonga, Cook Islands, Ovalau Island, Fiji, and Papua New Guinea (PNG), and their feces were examined for coccidians. Species included 5 seaside skinks (Emoia atrocostata), 1 Pacific blue-tailed skink (Emoia caeroleocauda), 2 Fiji slender treeskinks (Emoia concolor), 15 white-bellied copper-striped skinks (Emoia cyanura), 1 Bulolo River forest skink (Emoia guttata), 6 dark-bellied copper-striped skinks (Emoia impar), 5 Papua five-striped skinks (Emoia jakati), 2 Papua slender treeskinks (Emoia kordoana), 3 Papua robust treeskinks (Emoia longicauda), 1 brown-backed forest skink (Emoia loveridgei), 3 Papua black-sided skinks (Emoia pallidiceps), 2 Papua white-spotted skinks (Emoia physicae), 2 Papua yellow-head skinks (Emoia popei), 1 Papua brown forest skink (Emoia submetallica), and 5 Fiji barred treeskinks (Emoia trossula) Species of Eimeria (Ei.) were detected from these Emoia (Em.) spp. and are described here as new. Oocysts of Eimeria iovai n. sp. from Em. pallidiceps from PNG were ellipsoidal with a bilayered wall (L × W) 26.5 × 18.1 μm, with a length/width ratio (L/W) of 1.1. Both micropyle and oocyst residuum were absent, but a fragmented polar granule was present. This eimerian also was found in Em. atrocostata from PNG. Oocysts of Eimeria kirkpatricki n. sp. from Em. atrocostata from PNG were ellipsoidal with a bilayered wall, 18.6 × 13.5 μm, L/W 1.4. A micropyle and oocyst residuum were absent, but a fragmented polar granule was present. This eimerian was also shared by Em. cyanura from the Cook Islands and Fiji, Em. imparfrom the Cook Islands, Em. loveridgei from PNG, Em. pallidicepsfrom PNG, Em. popei from PNG, and Em. submetallica from PNG. Oocysts of Eimeria stevejayuptoni n. sp. from Em. longicaudawere subspheroidal to ellipsoidal with a bilayered wall, 18.7 × 16.6 μm, L/W 1.1. A

  9. Host range characteristics of the primate coccidian, Isospora arctopitheci Rodhain 1933 (Protozoa: Eimeriidae).

    Science.gov (United States)

    Hendricks, L D

    1977-02-01

    Studies were conducted on 35 primates, 12 carnivores, and 2 marsupials to determine their susceptibility to the primate coccidian, Isospora arctopitheci. Patent oocyst infections resulted in 12 of the 14 species of animals investigated. These included 6 genera of New World primates native to Panama: Saguinus geoffroyi, Aotus trivirgatus, Ateles fusciceps, Cebus capucinus, Alouatta villosa, and Saimiri sciureus. In addition 4 families of carnivores (2 domestic and 2 sylvatic) and 1 species of marsupial became infected following experimental exposure. These animals are represented respectively by the following 6 genera and species: Canis familiaris; Felis catus; Nasua nasua, and Potos flavus; Eiria barbara; and Didelphis marsupialis. Four Old World rhesus monkeys, Macaca mulatta, and 1 carnivore, Bassaricyon gabbii, did not become oocyst positive. This unusually large host range makes this isosporan unique among the coccidia that have been investigated to date.

  10. Caryospora neofalconis and other enteroparasites in raptors from Mexico

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    G. Santana-Sánchez

    2015-12-01

    Full Text Available A coprological survey of enteroparasites in raptors (60 Falconiformes from Central Mexico is reported. Three samples contained coccidian unsporulated oocysts, one contained Eimeria sp., one contained trematode eggs and one contained capillarid and trematode eggs and Eimeria sp. After sporulation at the laboratory, oocysts from a Falco peregrinus were identified as Caryospora neofalconis. The phylogenetic analysis of the C. neofalconis (GenBank accession number KT037081 showed a close relationship to the Australian strain RY 2014 isolate 16710 (GenBank accession number KJ634019 of Caryospora daceloe, with 99.2% similarity. As far as we are aware, this is the first report of C. neofalconis in raptors from Mexico and the Americas.

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

    Science.gov (United States)

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

    2013-07-01

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

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

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    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.

  13. Some new species of Caryospora (Apicomplexa: Eimeriidae from brazilian snakes, and a re-description of C. jararacae Carini, 1939

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    Ralph Lainson

    1991-09-01

    Full Text Available The mature ooxysts of six new species of Caryospora are described from the faeces of Brazilian snakes. They are differentiated from other species previously recorded from reptiles, largely on the size and shape of the oocyst and sporocyst, structure of the oocyst wall, and presence or absence of a polar body. C. paraensis n. sp., and C. carajasensis n. sp., are from the "false coral", Oxyrhopus petola digitalis; C. pseustesi n. sp., from the "egg-eater", Pseustes sulphureus sulphureus; C. epicratesi n. sp., from the "red boa", Epicrates cenchria cenchria; and C. micruri n. sp., and C. constancieae n. sp., from the "coral snake", Micrurus spixii spixii. A re-description is given of C. jararacae Carini, 1939, from the "jararaca" Bothrops atrox, embodying some additional morphological features.

  14. Lacteal immunity to enteric cryptosporidiosis in mice: immune dams do not protect their suckling pups.

    Science.gov (United States)

    Moon, H W; Woodmansee, D B; Harp, J A; Abel, S; Ungar, B L

    1988-03-01

    The susceptibilities of passively immunized principal and nonimmunized control suckling mice to orogastric challenge with Cryptosporidium parvum oocysts were compared. Principals were suckled by dams that had recovered from C. parvum infection. Controls were suckled by dams reared free of C. parvum infection. Principals and controls were equally susceptible to challenge. Principals were susceptible even when their dams were hyperimmunized by oral and parenteral booster inoculations with C. parvum oocysts. Immune dams produced serum antibody against C. parvum, while nonimmune dams did not. Anti-cryptosporidia immunoglobulin G (IgG) and IgA were demonstrated in whey extracted from the stomachs of principals that had suckled immune dams but not in whey extracted from the stomachs of controls. It was concluded that passive lacteal immunity is not an efficient means of protection against cryptosporidiosis in mice. As in other coccidian infections, protective immunity against cryptosporidiosis may depend more on immune cells than on antibody.

  15. The effects of salinomycin and roxarsone on the performance of broilers when included in the feed for four, five, or six weeks and infected with Eimeria species during the starter or grower phase of production.

    Science.gov (United States)

    Chapman, H D; Marsler, P; LaVorgna, M W

    2004-05-01

    The timing and magnitude of a coccidian infection, judged by the numbers of oocysts of Eimeria present in the litter, were affected by the duration of medication. In birds medicated for 6 wk and infected at 35 d of age, fewer oocysts were produced than in birds medicated for 4 or 5 wk whether infected at 18 or 35 d of age. Feed conversion at 6, 7, and 8 wk of birds infected at 18 d and medicated for 6 wk was less than that of birds medicated for 4 or 5 wk. Birds infected at 35 d and medicated for 6 wk had a lower feed conversion than birds medicated for 5 wk. Immunity to Eimeria tenella had developed by 8 wk in birds medicated for 4, 5, or 6 wk if infected at 18 d of age. Immunity did not develop in those birds medicated for 6 wk when infected at 35 d.

  16. Molecular detection of field isolates of Turkey Eimeria by polymerase chain reaction amplification of the cytochrome c oxidase I gene.

    Science.gov (United States)

    Rathinam, T; Gadde, U; Chapman, H D

    2015-07-01

    Oocysts of Eimeria spp. were isolated from litter samples obtained from 30 commercial turkey farms. Genomic DNA was extracted from clean oocysts, and polymerase chain amplification of the species-specific cytochrome c oxidase subunit I (COI) gene was performed for five species of turkey Eimeria. The species tested were Eimeria adenoeides, Eimeria meleagrimitis, Eimeria meleagridis, Eimeria dispersa, and Eimeria gallopavonis. All DNA samples were positive for E. meleagrimitis, nine were positive for E. adenoeides, two were positive for E. dispersa, and none for E. meleagridis and E. gallopavonis. E. meleagrimitis occurred as a single species in 21 (70 %) of the farms while 9 (30 %) farms had a mixed species with E. meleagrimitis and E. adenoeides and 2 (7 %) were triple positive with E. meleagrimitis, E. adenoeides, and E. dispersa. This is the first account of the field prevalence of turkey Eimeria species using molecular methods.

  17. Eimeria spp. from Japanese quails (Coturnix japonica: new characteristic features and diagnostic tools

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    Bruno P. Berto

    2013-12-01

    Full Text Available The Japanese quail Coturnix japonica originated from North Africa, Europe and Asia, is used worldwide as an experimental animal and model for aviculture. The current paper characterizes Eimeria bateri, Eimeria tsunodai and Eimeria uzura recovered from C. japonica. Based on the fact that quails have a global distribution, as are their coccidia, the findings of this study should provide the means for diagnosis of those Eimeria spp. in other regions and continents. Eimeria bateri showed the greatest intensity of infection and shed oocysts from the fourth day after infection; in contrast, E. tsunodai and E. uzura shed oocysts from the fifth day after infection. The three species shared a high degree of similarity and were all polymorphic. Yet, the application of line regressions, histograms and ANOVA provided means for the identification of these species. Finally, the algorithm was very efficient since verified that resultant values were not superimposed.

  18. Study on impression smears of hepatic coccidiosis in rabbits.

    Science.gov (United States)

    Sivajothi, S; Reddy, B Sudhakara; Rayulu, V C

    2016-09-01

    Hepatic coccidiosis is a contagious and lethal disease condition in rabbits. The disease was recorded in six rabbits suffering with watery diarrhoea. Clinically, affected rabbits showed decreased growth rate, anorexia, debilitation, diarrhea and rough hair coat. Examination of the faecal samples revealed the presence of unsporulated oocysts of Eimeria spp. After sporulation Eimeria stiedae oocysts were identified. Postmortem examination revealed hepatomegaly with presence of discrete yellowish-white nodules on the surface of the liver. Impression smears from the liver revealed the presence of numerous developmental stages of E. stiedae corresponding with the stage of the liver lesion and also represent the histological changes of the liver. Rabbits were treated with a combination of sulphaquinoxaline and diaveridine for five days.

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

    Science.gov (United States)

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

    2011-12-01

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

  20. First survey of endoparasites in pet ferrets in Italy.

    Science.gov (United States)

    d'Ovidio, D; Pepe, P; Ianniello, D; Noviello, E; Quinton, Jean-Francois; Cringoli, G; Rinaldi, L

    2014-06-16

    Endoparasites are infrequently reported in ferrets. A cross-sectional survey was conducted to determine the prevalence of intestinal parasites in pet ferrets in southern Italy. Fresh fecal samples were randomly collected from 50 ferrets housed in pet shops or privately owned. All fecal samples were processed using the FLOTAC pellet technique to identify and count helminthic eggs/larvae and protozoan cysts/oocysts. In addition, the samples were analyzed also by the Remel XpectGiardia/Cryptosporidium immunoassay. Intestinal parasites were detected in 15 out of 50 ferrets (30%). Eggs of ancylostomids were found in 28.0% (14/50) of the animals and oocysts of Sarcocystis were detected in one ferret (2.0%). None of the samples was positive for Cryptosporidium or Giardia. To the authors' knowledge, this is the first report of sarcosporidiosis in a pet ferret in Italy.

  1. An Ancient Protein Phosphatase, SHLP1, Is Critical to Microneme Development in Plasmodium Ookinetes and Parasite Transmission

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    Eva-Maria Patzewitz

    2013-03-01

    Full Text Available Signaling pathways controlled by reversible protein phosphorylation (catalyzed by kinases and phosphatases in the malaria parasite Plasmodium are of great interest, for both increased understanding of parasite biology and identification of novel drug targets. Here, we report a functional analysis in Plasmodium of an ancient bacterial Shewanella-like protein phosphatase (SHLP1 found only in bacteria, fungi, protists, and plants. SHLP1 is abundant in asexual blood stages and expressed at all stages of the parasite life cycle. shlp1 deletion results in a reduction in ookinete (zygote development, microneme formation, and complete ablation of oocyst formation, thereby blocking parasite transmission. This defect is carried by the female gamete and can be rescued by direct injection of mutant ookinetes into the mosquito hemocoel, where oocysts develop. This study emphasizes the varied functions of SHLP1 in Plasmodium ookinete biology and suggests that it could be a novel drug target for blocking parasite transmission.

  2. The first detection of Toxoplasma gondii DNA in environmental fruits and vegetables samples.

    Science.gov (United States)

    Lass, A; Pietkiewicz, H; Szostakowska, B; Myjak, P

    2012-06-01

    Toxoplasma gondii infections are prevalent in humans and animals all over the world. The aim of the study was to estimate the occurrence of T. gondii oocysts in fruits and vegetables and determine the genotype of the parasites. A total number of 216 fruits and vegetables samples were taken from shops and home gardens located in the area of northern Poland. Oocysts were recovered with the flocculation method. Then, real-time polymerase chain reaction (PCR) targeting the B1 gene was used for specific T. gondii detection and quantification. Toxoplasma DNA was found in 21 samples. Genotyping at the SAG2 locus showed SAG2 type I and SAG2 type II. This is the first investigation describing T. gondii DNA identification in a large number of fruits and vegetables samples with rapid molecular detection methods. The results showed that fruits and vegetables contaminated with T. gondii may play a role in the prevalence of toxoplasmosis in Poland.

  3. Laminin and the malaria parasite's journey through the mosquito midgut.

    Science.gov (United States)

    Arrighi, Romanico B G; Lycett, Gareth; Mahairaki, Vassiliki; Siden-Kiamos, Inga; Louis, Christos

    2005-07-01

    During the invasion of the mosquito midgut epithelium, Plasmodium ookinetes come to rest on the basal lamina, where they transform into the sporozoite-producing oocysts. Laminin, one of the basal lamina's major components, has previously been shown to bind several surface proteins of Plasmodium ookinetes. Here, using the recently developed RNAi technique in mosquitoes, we used a specific dsRNA construct targeted against the LANB2 gene (laminin gamma1) of Anopheles gambiae to reduce its mRNA levels, leading to a substantial reduction in the number of successfully developed oocysts in the mosquito midgut. Moreover, this molecular relationship is corroborated by the intimate association of developing P. berghei parasites and laminin in the gut, as observed using confocal microscopy. Our data support the notion of laminin playing a functional role in the development of the malaria parasite within the mosquito midgut.

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

    Science.gov (United States)

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

    2016-10-01

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

  5. Caryospora peneireiroi n. sp. (Apicomplexa: Eimeriidae) in the common kestrel, Falco tinnunculus (Falconiformes: Falconidae), in mainland Portugal.

    Science.gov (United States)

    Cardozo, Sergian Vianna; Berto, Bruno Pereira; Caetano, Inês; Maniero, Viviane Camara; Fonseca, Isabel Pereira da; Lopes, Carlos Wilson Gomes

    2016-06-07

    The common kestrel Falco tinnunculus Linnaeus, 1758, is a widespread raptor, native in Europe, Asia and Africa, and vagrant in the Americas. In the current work, 27 fecal samples were collected from common kestrels kept in the Lisbon Center for Wild Animal Recovery, located at Monsanto Forest Park, Lisbon, Portugal. Five (19%) of them were found to be passing an undescribed species of Caryospora in their feces. The oocysts of Caryospora peneireiroi n. sp. were ellipsoidal with a bilayered wall and measured 47.1 × 37.6 µm with a shape index of 1.25. No micropyle, oocyst residuum or polar granule was present. The sporocysts were subspherical, measuring 25.1 × 24.3 µm. Stieda, sub-Stieda and para-Stieda bodies were absent. The sporocyst residuum was composed of many homogenous globules scattered throughout the periphery of the sporocyst. This is the fourth caryosporan species reported from F. tinnunculus.

  6. Caryospora peneireiroi n. sp. (Apicomplexa: Eimeriidae in the common kestrel, Falco tinnunculus (Falconiformes: Falconidae, in mainland Portugal

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    Sergian Vianna Cardozo

    2016-01-01

    Full Text Available Abstract The common kestrel Falco tinnunculus Linnaeus, 1758, is a widespread raptor, native in Europe, Asia and Africa, and vagrant in the Americas. In the current work, 27 fecal samples were collected from common kestrels kept in the Lisbon Center for Wild Animal Recovery, located at Monsanto Forest Park, Lisbon, Portugal. Five (19% of them were found to be passing an undescribed species of Caryospora in their feces. The oocysts of Caryospora peneireiroi n. sp. were ellipsoidal with a bilayered wall and measured 47.1 × 37.6 µm with a shape index of 1.25. No micropyle, oocyst residuum or polar granule was present. The sporocysts were subspherical, measuring 25.1 × 24.3 µm. Stieda, sub-Stieda and para-Stieda bodies were absent. The sporocyst residuum was composed of many homogenous globules scattered throughout the periphery of the sporocyst. This is the fourth caryosporan species reported from F. tinnunculus.

  7. A new coccidian, Isospora rheae sp. nov. (Apicomplexa, Eimeriidae, from Rhea americana (Aves, Rheidae from South America

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    Samira S.M. Gallo

    2014-12-01

    Full Text Available A new species of coccidian (Protozoa: Apicomplexa: Eimeriidae obtained from rheas, Rhea americana, is reported in Brazil. Oocysts of Isospora rheae sp. nov. are spherical to subspheroidal, measuring 22.6 × 21.0 µm, and have a double and smooth wall that is approximately 1.7 µm thick. The micropyle, oocyst residuum and polar granule are absent. Sporocysts are slightly ovoid, measuring 13.9 × 9.6 µm. The Stieda body is flattened, the substieda body is pointed, irregular and wavy and the sporocyst residuum is composed of scattered granules of varying sizes. Sporozoites have an oblong refractile body and one nucleus. This is the first description of an isosporid coccidian infecting birds of the family Rheidae.

  8. The aromatic amino acid hydroxylase genes AAH1 and AAH2 in Toxoplasma gondii contribute to transmission in the cat.

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    Zi T Wang

    2017-03-01

    Full Text Available The Toxoplasma gondii genome contains two aromatic amino acid hydroxylase genes, AAH1 and AAH2 encode proteins that produce L-DOPA, which can serve as a precursor of catecholamine neurotransmitters. It has been suggested that this pathway elevates host dopamine levels thus making infected rodents less fearful of their definitive Felidae hosts. However, L-DOPA is also a structural precursor of melanins, secondary quinones, and dityrosine protein crosslinks, which are produced by many species. For example, dityrosine crosslinks are abundant in the oocyst walls of Eimeria and T. gondii, although their structural role has not been demonstrated, Here, we investigated the biology of AAH knockout parasites in the sexual reproductive cycle within cats. We found that ablation of the AAH genes resulted in reduced infection in the cat, lower oocyst yields, and decreased rates of sporulation. Our findings suggest that the AAH genes play a predominant role during infection in the gut of the definitive feline host.

  9. Prevalence, pathology and risk factors for coccidiosis in domestic rabbits (Oryctolagus cuniculus) in selected regions in Kenya.

    Science.gov (United States)

    Okumu, P O; Gathumbi, P K; Karanja, D N; Mande, J D; Wanyoike, M M; Gachuiri, C K; Kiarie, N; Mwanza, R N; Borter, D K

    2014-01-01

    The potential for rabbit production in Kenya is high. However, high morbidity and mortality of domestic rabbits were reported. The aim of the study was to determine the pathology, prevalence and the predisposing factors to coccidiosis in domestic rabbits in selected regions in Kenya. A total of 61 farms keeping rabbits in six different counties were visited in the survey. A total of 2680 live rabbits were examined and 61 rabbits and 302 fecal samples were randomly collected from the farms and examined for coccidian oocysts by ante-mortem and post-mortem methods. The predisposing factors to coccidiosis were assessed through questionnaires and direct observation. Chi square (χ(2)) statistics was used with P values coccidiosis and hepatic coccidiosis was 29.5% and 11.5%, respectively. Higher counts of coccidian oocysts per gram of feces were recovered in weaners than in growers and adults rabbits (P coccidiosis in domestic rabbits in Kenya.

  10. Caryospora neofalconis and other enteroparasites in raptors from Mexico

    Science.gov (United States)

    Santana-Sánchez, G.; Flores-Valle, I.T.; González-Gómez, M.; Vega-Sánchez, V.; Salgado-Miranda, C.; Soriano-Vargas, E.

    2015-01-01

    A coprological survey of enteroparasites in raptors (60 Falconiformes) from Central Mexico is reported. Three samples contained coccidian unsporulated oocysts, one contained Eimeria sp., one contained trematode eggs and one contained capillarid and trematode eggs and Eimeria sp. After sporulation at the laboratory, oocysts from a Falco peregrinus were identified as Caryospora neofalconis. The phylogenetic analysis of the C. neofalconis (GenBank accession number KT037081) showed a close relationship to the Australian strain RY 2014 isolate 16710 (GenBank accession number KJ634019) of Caryosporadaceloe, with 99.2% similarity. As far as we are aware, this is the first report of C. neofalconis in raptors from Mexico and the Americas. PMID:26543806

  11. Immune responses and protective effect in mice vaccinated orally with surface sporozoite protein of Eimeria falciformis in ISCOMs.

    Science.gov (United States)

    Kazanji, M; Laurent, F; Péry, P

    1994-07-01

    Immunostimulating complexes (ISCOMs) were built after treatment of a purified surface protein from Eimeria falciformis sporozoites with a palmitic acid derivation, leading to a high ratio (33-64%) of P27 incorporation in these cage-like structures. P27 kept its antigenicity after incorporation in ISCOMs, which induced, after iterative intubations by the oral route to groups of mice, a systemic IgG response, a local IgA response, and a local enhanced cellular response as demonstrated by lymphoproliferation of mesenteric lymph node cells upon in vitro stimulation with antigen. This immunization (120 micrograms in six oral doses at 2-day intervals) afforded mice a partial protection (60%) against a subsequent 400 oocyst challenge. The reduction in daily oocyst excretion was corroborated by significantly different weight losses between immunized and control mice on days 9 and 10 postinfection and the subsequent death of these control mice. These observations provide the first application of ISCOMs to parasitic intestinal diseases.

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

    Science.gov (United States)

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

    2016-01-01

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

  13. A new species of Caryospora (Apicomplexa: Eimeriidae) from the bald eagle, Haliaeetus leucocephalus (Accipitriformes: Accipitridae), from Kansas.

    Science.gov (United States)

    McAllister, Chris T; Duszynski, Donald W; McKown, Richard D

    2013-04-01

    Between March 1989 and February 1994, 4 bald eagles ( Haliaeetus leucocephalus ) from various localities in Kansas were examined for coccidia. One (25%) of the bald eagles was found to be passing an undescribed species of Caryospora in its feces. Oocysts of Caryospora hanebrinki n. sp. are ellipsoidal to ovoidal with a bilayered wall and measure 48.1 × 42.1 μm with a shape index of 1.2. A micropyle, oocyst residuum, and polar granule were absent. Sporocysts are spheroidal, 24.8 μm wide. Stieda, substieda, and parastieda bodies were absent; a spheroidal sporocyst residuum is present; it measures 17.5 μm and is composed of many intact homogenous globules with a few dispersed in a loose spiral around the sporocysts. This is the first caryosporan documented from the bald eagle and is the largest known Caryospora from raptors.

  14. The Effect of Selected Dietary Supplements on the Occurrence of Coccidia in the Alimentary Tract of Chicken

    Directory of Open Access Journals (Sweden)

    Luboš Zábranský

    2013-10-01

    Full Text Available Poultry farming represents 30 % of meat production worldwide. The occurrence of animal parasites is an enormous problem of poultry farming. This problem is recorded from factory farming as well as from small farming and causes financial losses. The aim of this study was to gain basic data about the occurrence of coccidia in the intestines and about the occurrence of coccidia oocysts in the excrements of indoor broilers and carrier hens. Broilers and carrier hens were fed by dietary supplements which were supposed to have influence on native gut microflora composition and on weight gain. The experiment was conducted on 250 individuals of broilers. Broilers were divided into 5 groups according to dietary supplement (one group consisted of 50 individuals. Experimental groups were as follows: prebiotics, probiotics, homeopathic, coccidiostatics. One control group was also made. In the experimental groups dietary supplements were served every day from the 8th to 22nd day after the birth. The samples of excrements were tested in a laboratory twice a week. During 42nd and 57th day after the birth the samples of intestines of death chicken were tested three times (five individuals was chosen from every experimental group. Every individual was weighted before the slaughter, the samples of intestines were screened for occurrence, amount and type of coccidia right after the slaughter. The decrease of frequency of coccidia oocysts occurrence in the excrements in the experimental group prebiotics, homeopathics and control group in comparison with experimental group probiotics and coccidiostats was proved. The frequency of coccidia oocysts occurrence was the lowest by the experimental group prebiotics and by control group. The frequency of coccidia oocysts occurrence was the highest by the experimental group probiotics and coccidiostats. The highest weight gains were recorded by the control group (ø2.55 in comparison with the group with coccidiostats (ø 1.99.

  15. Use of Artemisia annua as a natural coccidiostat in free-range broilers and its effects on infection dynamics and performance.

    Science.gov (United States)

    de Almeida, Gustavo F; Horsted, Klaus; Thamsborg, Stig M; Kyvsgaard, Niels C; Ferreira, Jorge F S; Hermansen, John E

    2012-05-25

    This work investigated the preventive effect of Artemisia annua L. dried leaves supplied as a botanical coccidiostat to two broiler genotypes reared in a Danish free-range system in a factorial experiment (two genotypes and ± supplement of dried A. annua leaves). The genotypes White Bresse L40, a pure slow-growing line, and Kosmos 8 Ross, a hybrid genotype with medium growing characteristics, were used. Broilers were raised indoor until 29-days-old and kept free of parasites. Twelve groups of 30 randomly selected broilers were placed in the range forming three replicates for each treatment combination. The paddocks were cultivated with a mix of grass and clover. A separate group of broilers was naturally infected with Eimeria spp. oocysts and five animals nominated as "seeders" were introduced to the above mentioned 12 groups, 10 days after its formation, with each group consisting of 35 animals per plot. This infection strategy was meant to imitate the transmission pathway observed at farm level. Ten individual birds from each of the 12 groups, in total 120 animals of mixed sex, were monitored twice weekly for 30 days for oocysts excretion. PCR of pooled faecal samples, oocyst morphology and localization upon necropsy were used to identify the Eimeria species involved in the infection. In general, broilers from both genotypes in the range coped well with a coccidia infection caused by Eimeria acervulina and Eimeria maxima as no clinical symptoms, or deaths, were reported during the experiment. In general, broilers supplemented with A. annua dried leaves showed a significantly (ptreatment - accounted mainly for the fact that Kosmos females responded positively to the Artemisia treatment while Kosmos males responded negatively, and only minor differences were found between sexes for the White Bresse genotype. In conclusion, supply of A. annua dried leaves as a botanical coccidiostat significantly reduced oocyst output in free ranged broilers and thus may form part

  16. Changes in parasite transmission stage excretion after pheasant release

    OpenAIRE

    2006-01-01

    The production of parasite transmission stages was investigated in the faeces of 77 farm-bred ring-necked pheasants (Phasianus colchicus). Coccidian oocysts (Eimeria sp.), and nematode eggs (Heterakis sp., and Capillaria-like eggs) were recovered before and after release but all birds were treated prior to release. Treatment with fenbendazole significantly reduced the abundance of trans-mission-stage excretion for all parasites, and reduced the prevalence in the case of Eimeria sp. and Hetera...

  17. Secondary transmission of cryptosporidiosis associated with well water consumption: two case studies

    Directory of Open Access Journals (Sweden)

    Natania Carol Cavalcante Rezende

    2016-04-01

    Full Text Available Abstract: Cryptosporidiosis is a very prominent disease in the field of public health, and usually causes diarrhea. We describe two immunocompetent patients who presented with chronic diarrhea that was ultimately found to be caused by continuous exposure to well water contaminated with the microbial cysts (oocysts of the Cryptosporidium spp parasite. We describe the patients' histories and possible explanations for their prolonged symptoms.

  18. Toxoplasmosis – An update

    OpenAIRE

    Mittal, Veena; Ichhpujani, R L

    2011-01-01

    Toxplasmosis is an important zoonotic disease caused by protozoan parasite Toxoplasma gondii. The disease affects one-third of the total world population. Transmission of the disease is mainly by ingestion of food or water contaminated with oocysts. Congenital toxoplasmosis occurs from the transplacental passage of the parasite from mother to fetus. In most adults it does not cause serious illness, but it can cause blindness and mental retardation in congenitally infected children, and it is ...

  19. Anticoccidial effects of herbal extracts on Eimeria tenella infection in broiler chickens: in vitro and in vivo study.

    Science.gov (United States)

    Habibi, Hasan; Firouzi, Sobhan; Nili, Hasan; Razavi, Mostafa; Asadi, Seyedeh Leili; Daneshi, Sajad

    2016-06-01

    Safe alternative anticoccidial drug to chemical feed additives are herbal extracts, because they don't results to tissue residue and drug resistance. In order to evaluate the effects of herbal extracts to control avian coccidiosis, 180 one-day-old broiler chickens were randomly divided into nine equal groups, as follows: (1) Biarum bovei (2) Nectaroscordum tripedale( 3) Dorema aucheri (4) Cichorium intybus (5) Prangos ferulaceae (6) diclazuril (7) Artemisia absinthium (8) infected control (9) uninfected control (each contains two groups). Administration of herbal extracts and supplementation of diclazuril was began 2 days before challenge and lasted for the duration of the experiment. The chicks of all the groups except uninfected control group were inoculated orally with sporulated oocysts (3 × 10(3) oocysts of Eimeria tenella) on the day 22 of age. The criteria employed were: body weight, feed conversion ratio, blood in feces, survival rate, lesion scoring, number of oocyst output per gram feces and histopathological changes. For histopathological evaluation, on day 12 post inoculation three birds from each group were randomly selected and humanly sacrificed. N. tripedale and diclazuril revealed better results in terms of growth performance, lesion score, extent of bloody diarrhea and oocyst count as compared to other herbal extracts. The increase in the severity of lesions was observed in groups of D. aucheri, A. absinthium, B. bovei, P. ferulaceae, C. intybus, diclazuril and N. tripedale, respectively. In conclusion, the current study showed that herbal extracts were effective in control of coccidiosis caused by the E. tenella infection.

  20. 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..