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Sample records for eimeria tenella sporozoites

  1. The rhoptry proteome of Eimeria tenella sporozoites

    KAUST Repository

    Oakes, Richard D.; Kurian, Dominic; Bromley, Elizabeth V.; Ward, Chris; Lal, Kalpana; Blake, Damer P.; Reid, Adam James; Pain, Arnab; Sinden, Robert E.; Wastling, Jonathan M.; Tomley, F. M. M Fiona

    2013-01-01

    Proteins derived from the rhoptry secretory organelles are crucial for the invasion and survival of apicomplexan parasites within host cells. The rhoptries are club-shaped organelles that contain two distinct subpopulations of proteins that localise to separate compartments of the organelle. Proteins from the neck region (rhoptry neck proteins, RON) are secreted early in invasion and a subset of these is critical for the formation and function of the moving junction between parasite and host membranes. Proteins from the bulb compartment (rhoptry protein, ROP) are released later, into the nascent parasitophorous vacuole where they have a role in modifying the vacuolar environment, and into the host cell where they act as key determinants of virulence through their ability to interact with host cell signalling pathways, causing an array of downstream effects. In this paper we present the results of an extensive proteomics analysis of the rhoptry organelles from the coccidian parasite, Eimeria tenella, which is a highly pathogenic parasite of the domestic chicken causing severe caecal coccidiosis. Several different classes of rhoptry protein have been identified. First are the RON proteins that have varying degrees of similarity to proteins of Toxoplasma gondii and Neospora caninum. For some RON families, E. tenella expresses more than one gene product and many of the individual RON proteins are differentially expressed between the sporozoite and merozoite developmental stages. The E. tenella sporozoite rhoptry expresses only a limited repertoire of proteins with homology to known ROP proteins from other coccidia, including just two secreted ROP kinases, both of which appear to be equipped for catalytic activity. Finally, a large number of hitherto undescribed proteins that map to the sporozoite rhoptry are identified, many of which have orthologous proteins encoded within the genomes of T. gondii and N. caninum. © 2012 .

  2. The rhoptry proteome of Eimeria tenella sporozoites

    KAUST Repository

    Oakes, Richard D.

    2013-02-01

    Proteins derived from the rhoptry secretory organelles are crucial for the invasion and survival of apicomplexan parasites within host cells. The rhoptries are club-shaped organelles that contain two distinct subpopulations of proteins that localise to separate compartments of the organelle. Proteins from the neck region (rhoptry neck proteins, RON) are secreted early in invasion and a subset of these is critical for the formation and function of the moving junction between parasite and host membranes. Proteins from the bulb compartment (rhoptry protein, ROP) are released later, into the nascent parasitophorous vacuole where they have a role in modifying the vacuolar environment, and into the host cell where they act as key determinants of virulence through their ability to interact with host cell signalling pathways, causing an array of downstream effects. In this paper we present the results of an extensive proteomics analysis of the rhoptry organelles from the coccidian parasite, Eimeria tenella, which is a highly pathogenic parasite of the domestic chicken causing severe caecal coccidiosis. Several different classes of rhoptry protein have been identified. First are the RON proteins that have varying degrees of similarity to proteins of Toxoplasma gondii and Neospora caninum. For some RON families, E. tenella expresses more than one gene product and many of the individual RON proteins are differentially expressed between the sporozoite and merozoite developmental stages. The E. tenella sporozoite rhoptry expresses only a limited repertoire of proteins with homology to known ROP proteins from other coccidia, including just two secreted ROP kinases, both of which appear to be equipped for catalytic activity. Finally, a large number of hitherto undescribed proteins that map to the sporozoite rhoptry are identified, many of which have orthologous proteins encoded within the genomes of T. gondii and N. caninum. © 2012 .

  3. Assessment of humoral immunity to Eimeria tenella sporozoites in chickens by ELISA

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    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.

  4. In vitro inhibition of Eimeria tenella sporozoite invasion into host cells by probiotics.

    Science.gov (United States)

    Hessenberger, S; Schatzmayr, G; Teichmann, K

    2016-10-15

    The aim was to study the effects of probiotics isolated from the intestinal tract of livestock animals on Eimeria tenella invasion into Madin-Darby bovine kidney (MDBK) cells in vitro. E. tenella sporozoites were purified and labeled with 5(6)-carboxyfluorescein diacetate N-succinimidyl ester before seeding on cell cultures, and invasion was evaluated by fluorescence microscopy. Two protocols (A and B) were used. In protocol A, Enterococcus faecium # 589 or Lactobacillus salivarius subsp. salivarius # 505 were added together with sporozoites to MDBK cell cultures and invasion was evaluated after incubation for approximately 20h. Viable, dead, or spent culture supernatants of probiotics were tested. In protocol B, viable probiotics were incubated with MDBK cells for one hour before sporozoites were added and invasion was evaluated after two more hours of incubation. Parasite invasion of viable, dead, or spent culture supernatant of E. faecium # 589 was assessed. Using protocol A, it was shown that parasite invasion was inhibited by viable (80%) or dead (75%) E. faecium # 589. While inhibition by viable L. salivarius subsp. salivarius # 505 was not valid at the highest concentration and not significant at the other test concentrations, dead cells inhibited parasite invasion up to 45%. Spent culture supernatants of both probiotics had no influence on parasite invasion. Using protocol B, it was shown that viable Bifidobacterium animalis subsp. animalis # 503, E. faecium # 497, E. faecium # 589, L. reuteri # 514, L. salivarius subsp. salivarius # 505, and Bacillus subtilis # 588 inhibited parasite invasion into MDBK cells up to 80%. Anticoccidial activity was strain-specific for E. faecium strains, and the strongest effect was shown by E. faecium # 589. Anticoccidial effects of some of the tested probiotics have already been shown in vivo, which makes them candidates to prevent coccidiosis. These findings have now been confirmed in vitro. The used parasite invasion

  5. 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

  6. Identification of common immunodominant antigens of Eimeria tenella, Eimeria acervulina and Eimeria maxima by immunoproteomic analysis.

    Science.gov (United States)

    Liu, Lianrui; Huang, Xinmei; Liu, Jianhua; Li, Wenyu; Ji, Yihong; Tian, Di; Tian, Lu; Yang, Xinchao; Xu, Lixin; Yan, Ruofeng; Li, Xiangrui; Song, Xiaokai

    2017-05-23

    Clinical chicken coccidiosis is mostly caused by simultaneous infection of several Eimeria species, and host immunity against Eimeria is species-specific. It is urgent to identify common immunodominant antigen of Eimeria for developing multivalent anticoccidial vaccines. In this study, sporozoite proteins of Eimeria tenella, Eimeria acervulina and Eimeria maxima were analyzed by two-dimensional electrophoresis (2DE). Western bot analysis was performed on the yielded 2DE gel using antisera of E. tenella E. acervulina and E. maxima respectively. Next, the detected immunodominant spots were identified by comparing the data from MALDI-TOF-MS/MS with available databases. Finally, Eimeria common antigens were identified by comparing amino acid sequence between the three Eimeria species. The results showed that analysis by 2DE of sporozoite proteins detected 629, 626 and 632 protein spots from E. tenella, E. acervulina and E. maxima respectively. Western bot analysis revealed 50 (E. tenella), 64 (E. acervulina) and 57 (E. maxima) immunodominant spots from the sporozoite 2DE gels of the three Eimeria species. The immunodominant spots were identified as 33, 27 and 25 immunodominant antigens of E. tenella, E. acervulina and E. maxima respectively. Fifty-four immunodominant proteins were identified as 18 ortholog proteins among the three Eimeria species. Finally, 5 of the 18 ortholog proteins were identified as common immunodominant antigens including elongation factor 2 (EF-2), 14-3-3 protein, ubiquitin-conjugating enzyme domain-containing protein (UCE) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). In conclusion, our results not only provide Eimeria sporozoite immunodominant antigen map and additional immunodominant antigens, but also common immunodominant antigens for developing multivalent anticoccidial vaccines.

  7. Eimeria tenella: host specificity in gallinaceous birds.

    Science.gov (United States)

    Vetterling, J M

    1976-02-01

    Eight species representing 8 genera of gallinaceous birds were used: Alectoris graeca; Colinus virginianus; Coturnix coturnix; Gallus gallus; Meleagris gallopavo; Numidia meleagris; Pavo cristatus; Phasianus colchicus. Three week-old birds were dosed with sporulated oocysts of Eimeria tenella Beltsville strain. At 4, 24, 48, 72, 96, 120 and 144, and 168 hr after inoculation, 1-3 infected birds and uninoculated controls of each species were killed by cardiac exsanguination. Pieces of intestines were fixed and examined for stages of E. tenella as stained paraffin sections or indirect fluorescent antibody preparations. Oocyst counts were made in droppings collected for the first 6 days of the patent period. Sporozoites were found in the lamina propria of some birds of 5 species at 4 hr postinoculation, but no stages were found thereafter except in the breeds of G. gallus and A. gracea. At 144 and 168 hr postinoculation, a few macrogametes were found in the ceca of 2 A. gracea, but no oocysts were found in the feces. No statistical difference was found between the number of oocysts produced/bird in the breeds of G. gallus examined. It is evident from these observations the E. tenella did not complete its life cycle in several close phylogenetic relatives of G. gallus, even though in other studies this parasite was found to complete its life cycle in cell cultures derived from the same birds.

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

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    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.

  9. Induction of Protective Immunity against Eimeria tenella, Eimeria maxima, and Eimeria acervulina Infections Using Dendritic Cell-Derived Exosomes

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    Gallego, Margarita; Lee, Sung Hyen; Lillehoj, Hyun Soon; Quilez, Joaquin; Lillehoj, Erik P.; Sánchez-Acedo, Caridad

    2012-01-01

    This study describes a novel immunization strategy against avian coccidiosis using exosomes derived from Eimeria parasite antigen (Ag)-loaded dendritic cells (DCs). Chicken intestinal DCs were isolated and pulsed in vitro with a mixture of sporozoite-extracted Ags from Eimeria tenella, E. maxima, and E. acervulina, and the cell-derived exosomes were isolated. Chickens were nonimmunized or immunized intramuscularly with exosomes and subsequently noninfected or coinfected with E. tenella, E. maxima, and E. acervulina oocysts. Immune parameters compared among the nonimmunized/noninfected, nonimmunized/infected, and immunized/infected groups were the numbers of cells secreting Th1 cytokines, Th2 cytokines, interleukin-16 (IL-16), and Ag-reactive antibodies in vitro and in vivo readouts of protective immunity against Eimeria infection. Cecal tonsils, Peyer's patches, and spleens of immunized and infected chickens had increased numbers of cells secreting the IL-16 and the Th1 cytokines IL-2 and gamma interferon, greater Ag-stimulated proliferative responses, and higher numbers of Ag-reactive IgG- and IgA-producing cells following in vitro stimulation with the sporozoite Ags compared with the nonimmunized/noninfected and nonimmunized/infected controls. In contrast, the numbers of cells secreting the Th2 cytokines IL-4 and IL-10 were diminished in immunized and infected chickens compared with the nonimmunized/noninfected and the nonimmunized/infected controls. Chickens immunized with Ag-loaded exosomes and infected in vivo with Eimeria oocysts had increased body weight gains, reduced feed conversion ratios, diminished fecal oocyst shedding, lessened intestinal lesion scores, and reduced mortality compared with the nonimmunized/infected controls. These results suggest that successful field vaccination against avian coccidiosis using exosomes derived from DCs incubated with Ags isolated from Eimeria species may be possible. PMID:22354026

  10. Molecular Characterization and Immune Protection of a New Conserved Hypothetical Protein of Eimeria tenella.

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    Qi Zhai

    Full Text Available The genome sequences of Eimeria tenella have been sequenced, but >70% of these genes are currently categorized as having an unknown function or annotated as conserved hypothetical proteins, and few of them have been studied. In the present study, a conserved hypothetical protein gene of E. tenella, designated EtCHP559, was cloned using rapid amplification of cDNA 5'-ends (5'RACE based on the expressed sequence tag (EST. The 1746-bp full-length cDNA of EtCHP559 contained a 1224-bp open reading frame (ORF that encoded a 407-amino acid polypeptide with the predicted molecular weight of 46.04 kDa. Real-time quantitative PCR analysis revealed that EtCHP559 was expressed at higher levels in sporozoites than in the other developmental stages (unsporulated oocysts, sporulated oocysts and second generation merozoites. The ORF was inserted into pCold-TF to produce recombinant EtCHP559. Using western blotting, the recombinant protein was successfully recognized by rabbit serum against E. tenella sporozoites. Immunolocalization by using EtCHP559 antibody showed that EtCHP559 was mainly distributed on the parasite surface in free sporozoites and became concentrated in the anterior region after sporozoites were incubated in complete medium. The EtCHP559 became uniformly dispersed in immature and mature schizonts. Inhibition of EtCHP559 function using anti-rEtCHP559 polyclonal antibody reduced the ability of E. tenella sporozoites to invade host cells by >70%. Animal challenge experiments demonstrated that the recombinant EtCHP559 significantly increased the average body weight gain, reduced the oocyst outputs, alleviated cecal lesions of the infected chickens, and resulted in anticoccidial index >160 against E. tenella. These results suggest that EtCHP559 plays an important role in sporozoite invasion and could be an effective candidate for the development of a new vaccine against E. tenella.

  11. Ekstrak Sambiloto Menurunkan Patogenesitas Ookista Eimeria Tenella

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    Yulia Yellita

    2012-11-01

    Full Text Available Eimeria tenella is one of the nine of Eimeria species, a pathogenic intraseluler protozoa causing aviancoccidiosis. Infection was initiated by the ingestion of sporulated oocysts. The aim of this study was toinvestigate the effect of E. tenella oocyst incubation in methanol extract of Andrographis paniculata beforeinfection in broiler performance. This research used 115 broiler DOC (CP 707 devided into five groups,each group consisted of 23 broilers. The infection with 1x105 oocyst were done at the 14th day old of chicken.The 1st group was placebo (KN, while the 2nd group was infected with unincubated oocyst (KP, and theother three groups i.e. : 3rd, 4th, 5th were infected with incubated oocyst in A. paniculata extract for 2, 4, and6 hours, respectively. The number of oocysts in feces were counted on day 5th to 14th post-infection, theheterophile and macrophages were counted from caecum histology preparation, by slaughtered threechickens of each of groups on the day 0,3,6.9, and 14 post infection, and accretion body weight wasmeasured by weighing chickens per week to five-week old chickens. The results of this study indicated thatthe incubation period the sporulated oocyst in the extract of A.paniculata for six hours before infection,reduced the number of oocysts production in the feces, the number of inflammatory cells (macrophages andheterophile in the cecum, and increases body weight (gain. In conclusion A.paniculata extract decreasedthe pathogenisity of E.tenella oocyst, so the extract of A.paniculata has good potential as anticoccidia. Itis high likely that A. paniculata extract has a potential to be anticoccidia.

  12. Evaluation of the antigenicity and immunogenicity of Eimeria tenella ...

    African Journals Online (AJOL)

    Background: The development of vaccine to control coccidiosis caused by Eimeria tenella (E. tenella) in chickens is intensifying because of the increasing threat of drug resistance to anticoccidial agents. It is important, therefore, to develop a reliable standard method for the assessment of vaccine afficacy particularly ...

  13. Induction of protective immunity against Eimeria tenella, Eimeria necatrix, Eimeria maxima and Eimeria acervulina infections using multivalent epitope DNA vaccines.

    Science.gov (United States)

    Song, Xiaokai; Ren, Zhe; Yan, Ruofeng; Xu, Lixin; Li, Xiangrui

    2015-06-04

    Avian coccidiosis is mostly caused by mixed infection of several Eimeria species under natural conditions and immunity to avian coccidiosis is largely dependent on T-cell immune response. In this study, 14 T-cell epitope fragments from eight antigens of Eimeria tenella (E. tenella), Eimeria necatrix (E. necatrix), Eimeria maxima (E. maxima) and Eimeria acervulina (E. acervulina) were ligated with pVAX1 producing 14 monovalent DNA vaccines, respectively. Protective immunity of the monovalent DNA vaccines was assessed by in vivo challenge experiments and then four most protective fragments of each species were chosen to construct multivalent epitope DNA vaccines with or without chicken IL-2 as genetic adjuvant. Protective efficacies of the epitope DNA vaccines on chickens against E. tenella, E. necatrix, E. maxima and E. acervulina were evaluated. The results showed that the constructed multivalent epitope DNA vaccines significantly increased body weight gain, alleviated enteric lesions and reduced oocyst output of the infected birds. Especially, the multivalent epitope DNA vaccines of pVAX1-NA4-1-TA4-1-LDH-2-EMCDPK-1 and pVAX1-NA4-1-TA4-1-LDH-2-EMCDPK-1-IL-2 not only significantly increased body weight gain, alleviated enteric lesions and reduced oocyst output of the infected birds, but also resulted in anti-coccidial index (ACI) more than 170 against E. tenella, E. necatrix, E. maxima and E. acervulina, which indicated they could induce protective immunity against E. tenella, E. necatrix, E. maxima and E. acervulina. Our findings suggest the constructed multivalent epitope DNA vaccines are the potential candidate multivalent vaccines against mixed infection of Eimeria. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. Quantitative proteomic studies in resistance mechanisms of Eimeria tenella against polyether ionophores.

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    Thabet, Ahmed; Honscha, Walther; Daugschies, Arwid; Bangoura, Berit

    2017-05-01

    Polyether ionophores are widely used to treat and control coccidiosis in chickens. Widespread use of anticoccidials resulted in worldwide resistance. Mechanisms of resistance development and expansion are complex and poorly understood. Relative proteomic quantification using LC-MS/MS was used to compare sensitive reference strains (Ref-1, Ref-2) with putatively resistant and moderately sensitive field strains (FS-R, FS-mS) of Eimeria tenella after isotopic labelling with tandem mass tags (TMT). Ninety-seven proteins were identified, and 25 of them were regulated. Actin was significantly upregulated in resistant strains in comparison with their sensitive counterparts. On the other hand, microneme protein (MIC4) was downregulated in resistant strains. Optimization of labelling E. tenella sporozoites by TMT might identify further proteins that play a role in the obvious complex mechanism leading to resistance against Monensin.

  15. A Novel Vaccine Delivery Model of the Apicomplexan Eimeria tenella Expressing Eimeria maxima Antigen Protects Chickens against Infection of the Two Parasites.

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    Tang, Xinming; Liu, Xianyong; Yin, Guangwen; Suo, Jingxia; Tao, Geru; Zhang, Sixin; Suo, Xun

    2017-01-01

    Vaccine delivery is critical in antigen discovery and vaccine efficacy and safety. The diversity of infectious diseases in humans and livestock has required the development of varied delivery vehicles to target different pathogens. In livestock animals, previous strategies for the development of coccidiosis vaccines have encountered several hurdles, limiting the development of multiple species vaccine formulations. Here, we describe a novel vaccine delivery system using transgenic Eimeria tenella expressing immunodominant antigens of Eimeria maxima . In this delivery system, the immune mapped protein 1 of E. maxima (EmIMP1) was delivered by the closely related species of E. tenella to the host immune system during the whole endogenous life cycle. The overexpression of the exogenous antigen did not interfere with the reproduction and immunogenicity of transgenic Eimeria . After immunization with the transgenic parasite, we detected EmIMP1's and E. maxima oocyst antigens' specific humoral and cellular immune responses. In particular, we observed partial protection of chickens immunized with transgenic E. tenella against subsequent E. maxima infections. Our results demonstrate that the transgenic Eimeria parasite is an ideal coccidia antigen delivery vehicle and represents a new type of coccidiosis vaccines. In addition, this model could potentially be used in the development of malaria live sporozoite vaccines, in which antigens from different strains can be expressed in the vaccine strain.

  16. A Novel Vaccine Delivery Model of the Apicomplexan Eimeria tenella Expressing Eimeria maxima Antigen Protects Chickens against Infection of the Two Parasites

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    Tang, Xinming; Liu, Xianyong; Yin, Guangwen; Suo, Jingxia; Tao, Geru; Zhang, Sixin; Suo, Xun

    2018-01-01

    Vaccine delivery is critical in antigen discovery and vaccine efficacy and safety. The diversity of infectious diseases in humans and livestock has required the development of varied delivery vehicles to target different pathogens. In livestock animals, previous strategies for the development of coccidiosis vaccines have encountered several hurdles, limiting the development of multiple species vaccine formulations. Here, we describe a novel vaccine delivery system using transgenic Eimeria tenella expressing immunodominant antigens of Eimeria maxima. In this delivery system, the immune mapped protein 1 of E. maxima (EmIMP1) was delivered by the closely related species of E. tenella to the host immune system during the whole endogenous life cycle. The overexpression of the exogenous antigen did not interfere with the reproduction and immunogenicity of transgenic Eimeria. After immunization with the transgenic parasite, we detected EmIMP1’s and E. maxima oocyst antigens’ specific humoral and cellular immune responses. In particular, we observed partial protection of chickens immunized with transgenic E. tenella against subsequent E. maxima infections. Our results demonstrate that the transgenic Eimeria parasite is an ideal coccidia antigen delivery vehicle and represents a new type of coccidiosis vaccines. In addition, this model could potentially be used in the development of malaria live sporozoite vaccines, in which antigens from different strains can be expressed in the vaccine strain. PMID:29375584

  17. Anticoccidial efficacy testing: In vitro Eimeria tenella assays as replacement for animal experiments.

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    Thabet, Ahmed; Zhang, Runhui; Alnassan, Alaa-Aldin; Daugschies, Arwid; Bangoura, Berit

    2017-01-15

    Availability of an accurate in vitro assay is a crucial demand to determine sensitivity of Eimeria spp. field strains toward anticoccidials routinely. In this study we tested in vitro models of Eimeria tenella using various polyether ionophores (monensin, salinomycin, maduramicin, and lasalocid) and toltrazuril. Minimum inhibitory concentrations (MIC 95 , MIC 50/95 ) for the tested anticoccidials were defined based on a susceptible reference (Houghton strain), Ref-1. In vitro sporozoite invasion inhibition assay (SIA) and reproduction inhibition assay (RIA) were applied on sensitive laboratory (Ref-1 and Ref-2) and field (FS-1, FS-2, and FS-3) strains to calculate percent of inhibition under exposure of these strains to the various anticoccidials (%I SIA and%I RIA, respectively). The in vitro data were related to oocyst excretion, lesion scores, performance, and global resistance indices (GI) assessed in experimentally infected chickens. Polyether ionophores applied in the RIA were highly effective at MIC 95 against Ref-1 and Ref-2 (%I RIA ≥95%). In contrast, all tested field strains displayed reduced to low efficacy (%I RIA animal model (p89%) against all strains used in this study. However, adjusted GI (GI adj ) for toltrazuril-treated groups exhibited differences between reference and field strains which might indicate varying sensitivity. RIA is a suitable in vitro tool to detect sensitivity of E. tenella towards polyether ionophores, and may thus help to reduce, replace, or refine use of animal experimentation for in vivo sensitivity assays. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Dietary Curcuma longa enhances resistance against Eimeria maxima and Eimeria tenella infections in chickens

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    The effects of dietary supplementation with an organic extract of Curcuma longa on systemic and local immune responses to experimental Eimeria maxima and E. tenella infections were evaluated in commercial broiler chickens. Infected chickens given the C. longa-containing diet had increased body weig...

  19. Embryonated chicken eggs as an alternative model for mixed Clostridium perfringens and Eimeria tenella infection in chickens.

    Science.gov (United States)

    Alnassan, Alaa Aldin; Shehata, Awad Ali; Kotsch, Marianne; Lendner, Matthias; Daugschies, Arwid; Bangoura, Berit

    2013-06-01

    The chorioallantoic membrane (CAM) of chicken embryo eggs is a suitable model for viral and bacterial infections. In the present study, a new approach for testing the pathogenesis and virulence of Clostridium perfringens and Eimeria tenella dual infections as a model using the CAM of embryonated chicken eggs was developed. For this purpose, 24 specific pathogen-free (SPF) embryonated chicken eggs were divided into four groups (n = 6) and designated group E, group CP, group CPE, and NC. Sporozoites of E. tenella (20,000 sporozoites) were inoculated into 10-day-old embryonated SPF chicken eggs (groups E and CPE) via allantoic sac route. At 15-day-old, eggs of groups CP and CPE were infected with 10 (4)  cfu C. perfringens via the same route. Assessment of pathogenicity was assessed using gross and histopathological lesions. Embryo mortality reached 17 % after mono-infection with C. perfringens and/or E. tenella and 50 % in the mixed-infected group. Lesions in the CAMs were most numerous and most severe in co-infected eggs (group CPE), reaching the maximum score of 3 in 50 % of the inoculated eggs (P < 0.01). In Eimeria spp.-infected eggs (group E), lesions of score were between 1 and 2. Mono-infection with C. perfringens did not lead to a significant occurrence of lesions. Histopathological investigations of the CAM revealed clusters of Gram-positive bacteria, infiltration with leukocytes, lymphocytes, and developmental stages of E. tenella in the co-infected group. These data suggest that embryonated eggs could be an in ovo model for studying the pathogenesis of mixed infection with Eimeria and C. perfringens.

  20. Identification and characterization of secreted proteins in Eimeria tenella

    Science.gov (United States)

    Ramlee, Intan Azlinda; Firdaus-Raih, Mohd; Wan, Kiew-Lian

    2015-09-01

    Eimeria tenella is a protozoan parasite that causes coccidiosis, an economically important disease in the poultry industry. The characterization of proteins that are secreted by parasites have been shown to play important roles in parasite invasion and are considered to be potential control agents. In this study, 775 proteins potentially secreted by E. tenella were identified. These proteins were further filtered to remove mitochondrial proteins. Out of 763 putative secreted proteins, 259 proteins possess transmembrane domains while another 150 proteins have GPI (Glycosylphosphatidylinositol) anchors. Homology search revealed that 315 and 448 proteins have matches with known and hypothetical proteins in the database, respectively. Within this data set, previously characterized secretory proteins such as micronemes, rhoptry kinases and dense granules were detected.

  1. Molecular characterization and analysis of a novel protein disulfide isomerase-like protein of Eimeria tenella.

    Science.gov (United States)

    Han, Hongyu; Dong, Hui; Zhu, Shunhai; Zhao, Qiping; Jiang, Lianlian; Wang, Yange; Li, Liujia; Wu, Youlin; Huang, Bing

    2014-01-01

    Protein disulfide isomerase (PDI) and PDI-like proteins are members of the thioredoxin superfamily. They contain thioredoxin-like domains and catalyze the physiological oxidation, reduction and isomerization of protein disulfide bonds, which are involved in cell function and development in prokaryotes and eukaryotes. In this study, EtPDIL, a novel PDI-like gene of Eimeria tenella, was cloned using rapid amplification of cDNA ends (RACE) according to the expressed sequence tag (EST). The EtPDIL cDNA contained 1129 nucleotides encoding 216 amino acids. The deduced EtPDIL protein belonged to thioredoxin-like superfamily and had a single predicted thioredoxin domain with a non-classical thioredoxin-like motif (SXXC). BLAST analysis showed that the EtPDIL protein was 55-59% identical to PDI-like proteins of other apicomplexan parasites. The transcript and protein levels of EtPDIL at different development stages were investigated by real-time quantitative PCR and western blot. The messenger RNA and protein levels of EtPDIL were higher in sporulated oocysts than in unsporulated oocysts, sporozoites or merozoites. Protein expression was barely detectable in unsporulated oocysts. Western blots showed that rabbit antiserum against recombinant EtPDIL recognized only a native 24 kDa protein from parasites. Immunolocalization with EtPDIL antibody showed that EtPDIL had a disperse distribution in the cytoplasm of whole sporozoites and merozoites. After sporozoites were incubated in complete medium, EtPDIL protein concentrated at the anterior of the sporozoites and appeared on the surface of parasites. Specific staining was more intense and mainly located on the parasite surface after merozoites released from mature schizonts invaded DF-1 cells. After development of parasites in DF-1 cells, staining intensified in trophozoites, immature schizonts and mature schizonts. Antibody inhibition of EtPDIL function reduced the ability of E. tenella to invade DF-1 cells. These results

  2. Molecular characterization and analysis of a novel protein disulfide isomerase-like protein of Eimeria tenella.

    Directory of Open Access Journals (Sweden)

    Hongyu Han

    Full Text Available Protein disulfide isomerase (PDI and PDI-like proteins are members of the thioredoxin superfamily. They contain thioredoxin-like domains and catalyze the physiological oxidation, reduction and isomerization of protein disulfide bonds, which are involved in cell function and development in prokaryotes and eukaryotes. In this study, EtPDIL, a novel PDI-like gene of Eimeria tenella, was cloned using rapid amplification of cDNA ends (RACE according to the expressed sequence tag (EST. The EtPDIL cDNA contained 1129 nucleotides encoding 216 amino acids. The deduced EtPDIL protein belonged to thioredoxin-like superfamily and had a single predicted thioredoxin domain with a non-classical thioredoxin-like motif (SXXC. BLAST analysis showed that the EtPDIL protein was 55-59% identical to PDI-like proteins of other apicomplexan parasites. The transcript and protein levels of EtPDIL at different development stages were investigated by real-time quantitative PCR and western blot. The messenger RNA and protein levels of EtPDIL were higher in sporulated oocysts than in unsporulated oocysts, sporozoites or merozoites. Protein expression was barely detectable in unsporulated oocysts. Western blots showed that rabbit antiserum against recombinant EtPDIL recognized only a native 24 kDa protein from parasites. Immunolocalization with EtPDIL antibody showed that EtPDIL had a disperse distribution in the cytoplasm of whole sporozoites and merozoites. After sporozoites were incubated in complete medium, EtPDIL protein concentrated at the anterior of the sporozoites and appeared on the surface of parasites. Specific staining was more intense and mainly located on the parasite surface after merozoites released from mature schizonts invaded DF-1 cells. After development of parasites in DF-1 cells, staining intensified in trophozoites, immature schizonts and mature schizonts. Antibody inhibition of EtPDIL function reduced the ability of E. tenella to invade DF-1 cells

  3. Identification and characterization of Eimeria tenella apical membrane antigen-1 (AMA1.

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    Lianlian Jiang

    Full Text Available Apical membrane antigen-1 (AMA1 is a micronemal protein of apicomplexan parasites that appears to be essential during the invasion of host cells. In this study, a full-length cDNA of AMA1 was identified from Eimeria tenella (Et using expressed sequence tag and the rapid amplification of cDNA ends technique. EtAMA1 had an open reading frame of 1608 bp encoding a protein of 535 amino acids. Quantitative real-time PCR analysis revealed that EtAMA1 was expressed at higher levels in sporozoites than in the other developmental stages (unsporulated oocysts, sporulated oocysts and second-generation merozoites. The ectodomain sequence was expressed as recombinant EtAMA1 (rEtAMA1 and rabbit polyclonal antibodies raised against the rEtAMA1 recognized a 58-kDa native parasite protein by Western Blotting and had a potent inhibitory effect on parasite invasion, decreasing it by approximately 70%. Immunofluorescence analysis and immunohistochemistry analysis showed EtAMA1 might play an important role in sporozoite invasion and development.

  4. Eimeria tenella: in vitro development in irradiated bovine kidney cells

    Energy Technology Data Exchange (ETDEWEB)

    Crane, M.St.J.; Schmatz, D.M.; Stevens, S.; Habbersett, M.C.; Murray, P.K. (Merck Sharp and Dohme Research Labs., Rahway, NJ (USA))

    1984-06-01

    The initial infection and first-generation development of Eimeria tenella was quantified using a cloned MDBK (Madin-Darby Bovine Kidney) cell line, irradiated with gamma radiation prior to infection, as the host cell. Irradiated cell cultures were found to be more susceptible to infection and had a greater capacity to support parasite development than non-irradiated cultures. It was suggested that the larger proportion of cells in the G/sub 2/ phase of the cell cycle, the larger individual cell size and the inhibition of cell division in the irradiated cultures were all factors contributing to the increased susceptibility to infection and capacity to support parasite growth and development. The application of this technique (host cell irradiation) to the cultivation of other intracellular, protozoan parasites is discussed.

  5. Eimeria tenella: in vitro development in irradiated bovine kidney cells

    International Nuclear Information System (INIS)

    Crane, M. St.J.; Schmatz, D.M.; Stevens, S.; Habbersett, M.C.; Murray, P.K.

    1984-01-01

    The initial infection and first-generation development of Eimeria tenella was quantified using a cloned MDBK (Madin-Darby Bovine Kidney) cell line, irradiated with gamma radiation prior to infection, as the host cell. Irradiated cell cultures were found to be more susceptible to infection and had a greater capacity to support parasite development than non-irradiated cultures. It was suggested that the larger proportion of cells in the G 2 phase of the cell cycle, the larger individual cell size and the inhibition of cell division in the irradiated cultures were all factors contributing to the increased susceptibility to infection and capacity to support parasite growth and development. The application of this technique (host cell irradiation) to the cultivation of other intracellular, protozoan parasites is discussed. (author)

  6. Attenuate Eimeria Tenella parasite by gamma radiation in chicken vaccination

    International Nuclear Information System (INIS)

    El-atar, M.A.

    1998-01-01

    Mature occysts of eimeria tenella were attenuated by different doses of gamma radiation. The vitality, pathogenicity and immunogenicity of these occysts were examined by infecting one day old broiler chicks. The study revealed that the irradiated occysts lost pathogenicity by increasing radiation dose. To examine the immunogenicity of irradiated occysts, chickens were challenged 28 days post immunogenic infection. It was shown that the irradiated occycts kept their immunogenicity but this ability decreased when the irradiation dose was increased. Also, the number of vaccination doses as well as the level of irradiation were studied. Occysts irradiated with 15, 18, 20 Krad were used to vaccinate one-day old broiler chicks for one or two times, and seven-day old chicks for three times. High level of protection was observed as shown by disappeaeance of clinical signs or mortality in most vaccinated groups

  7. Dietary Curcuma longa enhances resistance against Eimeria maxima and Eimeria tenella infections in chickens.

    Science.gov (United States)

    Kim, Duk Kyung; Lillehoj, Hyun S; Lee, Sung Hyen; Jang, Seung I; Lillehoj, Erik P; Bravo, David

    2013-10-01

    The effects of dietary supplementation with an organic extract of Curcuma longa on systemic and local immune responses to experimental Eimeria maxima and Eimeria tenella infections were evaluated in commercial broiler chickens. Dietary supplementation with C. longa enhanced coccidiosis resistance as demonstrated by increased BW gains, reduced fecal oocyst shedding, and decreased gut lesions compared with infected birds fed a nonsupplemented control diet. The chickens fed C. longa-supplemented diet showed enhanced systemic humoral immunity, as assessed by greater levels of serum antibodies to an Eimeria microneme protein, MIC2, and enhanced cellular immunity, as measured by concanavalin A-induced spleen cell proliferation, compared with controls. At the intestinal level, genome-wide gene expression profiling by microarray hybridization identified 601 differentially expressed transcripts (287 upregulated, 314 downregulated) in gut lymphocytes of C. longa-fed chickens compared with nonsupplemented controls. Based on the known functions of the corresponding mammalian genes, the C. longa-induced intestinal transcriptome was mostly associated with genes mediating anti-inflammatory effects. Taken together, these results suggest that dietary C. longa could be used to attenuate Eimeria-induced, inflammation-mediated gut damage in commercial poultry production.

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

    Science.gov (United States)

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

  9. Vaccination with Eimeria tenella elongation factor-1α recombinant protein induces protective immunity against E. tenella and E. maxima infections.

    Science.gov (United States)

    Lin, Rui-Qing; Lillehoj, Hyun S; Lee, Seung Kyoo; Oh, Sungtaek; Panebra, Alfredo; Lillehoj, Erik P

    2017-08-30

    Avian coccidiosis is caused by multiple species of the apicomplexan protozoan, Eimeria, and is one of the most economically devastating enteric diseases for the poultry industry worldwide. Host immunity to Eimeria infection, however, is relatively species-specific. The ability to immunize chickens against different species of Eimeria using a single vaccine will have a major beneficial impact on commercial poultry production. In this paper, we describe the molecular cloning, purification, and vaccination efficacy of a novel Eimeria vaccine candidate, elongation factor-1α (EF-1α). One day-old broiler chickens were given two subcutaneous immunizations one week apart with E. coli-expressed E. tenella recombinant (r)EF-1α protein and evaluated for protection against challenge infection with E. tenella or E. maxima. rEF-1α-vaccinated chickens exhibited increased body weight gains, decreased fecal oocyst output, and greater serum anti-EF-1α antibody levels following challenge infection with either E. tenella or E. maxima compared with unimmunized controls. Vaccination with EF-1α may represent a new approach to inducing cross-protective immunity against avian coccidiosis in the field. Published by Elsevier B.V.

  10. A genetic linkage map for the apicomplexan protozoan parasite Eimeria maxima and comparison with Eimeria tenella.

    Science.gov (United States)

    Blake, Damer P; Oakes, Richard; Smith, Adrian L

    2011-02-01

    Eimeria maxima is one of the seven Eimeria spp. that infect the chicken and cause the disease coccidiosis. The well characterised immunogenicity and genetic diversity associated with E. maxima promote its use in genetics-led studies on avian coccidiosis. The development of a genetic map for E. maxima, presented here based upon 647 amplified fragment length polymorphism markers typed from 22 clonal hybrid lines and assembled into 13 major linkage groups, is a major new resource for work with this parasite. Comparison with genetic maps produced for other coccidial parasites indicates relatively high levels of genetic recombination. Conversion of ∼14% of the markers representing the major linkage groups to sequence characterised amplified region markers can provide a scaffold for the assembly of future genomic sequences as well as providing a foundation for more detailed genetic maps. Comparison with the Eimeria tenella genetic map produced 10years ago has revealed a less biased marker distribution, with no more than nine markers mapped within any unresolved heritable unit. Nonetheless, preliminary bioinformatic characterisation of the three largest publicly available genomic E. maxima sequences suggest that the feature-poor/feature-rich structure which has previously been found to define the first sequenced E. tenella chromosome also defines the E. maxima genome. The significance of such a segmented genome and the apparent potential for variation in genetic recombination will be relevant to haplotype stability and the longevity of future anticoccidial strategies based upon multiple loci targeted by novel chemotherapeutic drugs or recombinant subunit vaccines. Copyright © 2010 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

  11. Ekstrak Sambiloto (Andrographis paniculata Menurunkan Jumlah Skizon, Mikrogamet, Makrogamet, dan Oosista Eimeria tenella (EXTRACT OF ANDROGRAPHIS PANICULATA DECREASED SCHIZONTS, MICROGAMETES, MACROGAMETES AND OOCYSTS NUMBER OF EIMERIA TENELLA

    Directory of Open Access Journals (Sweden)

    UMI CAHYANINGSIH

    2013-07-01

    Full Text Available The aim of this research was to observe the effect of ethanol extract of Andrographis paniculata givenin grading doses to the schizonts, microgamete, macrogamete, and oocytes counts of Eimeria tenella inchicken caecum. A total of ninety day old broiler chicks were used in the study. At two weeks old the broilerswere divided into six groups. Each group consisted of 15 broilers, the 6 groups were: (i negative control(broilers did not receive any treatment; (ii positive control (each animal were infected with 104 E. tenellaoocytes; (iii medicine control (each animal were infected with 104 E. tenella oocytes and coccidiostat; (ivA1 (each animal were infected with 104 E. tenella oocytes and paniculata extract 90 mg/kg body weight; (vA2 (each animal were infected with 104 E. tenella oocytes and paniculata extract 180 mg/kg body weight;and (vi A3 (each animal were infected with 104 E. tenella oocytes and paniculata extract 360 mg/kg bodyweight. At day 6, 9, 13, 16, and 22 post infection three broilers from each group were sacrificed and theirceca were collected for histopathological examination. The results showed that paniculata extract at dose90 mg/kg body weight and 180 mg/kg body weight was able to decrease the numbers of shizont, microgamete,macrogamete, and oocytes of E. tenella in the chicken caecum.

  12. Characterization of the Eimeria maxima sporozoite surface protein IMP1.

    Science.gov (United States)

    Jenkins, M C; Fetterer, R; Miska, K; Tuo, W; Kwok, O; Dubey, J P

    2015-07-30

    The purpose of this study was to characterize Eimeria maxima immune-mapped protein 1 (IMP1) that is hypothesized to play a role in eliciting protective immunity against E. maxima infection in chickens. RT-PCR analysis of RNA from unsporulated and sporulating E. maxima oocysts revealed highest transcription levels at 6-12h of sporulation with a considerable downregulation thereafter. Alignment of IMP1 coding sequence from Houghton, Weybridge, and APU-1 strains of E. maxima revealed single nucleotide polymorphisms that in some instances led to amino acid changes in the encoded protein sequence. The E. maxima (APU-1) IMP1 cDNA sequence was cloned and expressed in 2 different polyHis Escherichia coli expression vectors. Regardless of expression vector, recombinant E. maxima IMP1 (rEmaxIMP1) was fairly unstable in non-denaturing buffer, which is consistent with stability analysis of the primary amino acid sequence. Antisera specific for rEmaxIMP1 identified a single 72 kDa protein or a 61 kDa protein by non-reducing or reducing SDS-PAGE/immunoblotting. Immunofluorescence staining with anti-rEmaxIMP1, revealed intense surface staining of E. maxima sporozoites, with negligible staining of merozoite stages. Immuno-histochemical staining of E. maxima-infected chicken intestinal tissue revealed staining of E. maxima developmental stages in the lamnia propia and crypts at both 24 and 48 h post-infection, and negligible staining thereafter. The expression of IMP1 during early stages of in vivo development and its location on the sporozoite surface may explain in part the immunoprotective effect of this protein against E. maxima infection. Published by Elsevier B.V.

  13. Sensitive and specific identification by polymerase chain reaction of Eimeria tenella and Eimeria maxima, important protozoan pathogens in laboratory avian facilities.

    Science.gov (United States)

    Lee, Hyun-A; Hong, Sunhwa; Chung, Yungho; Kim, Okjin

    2011-09-01

    Eimeria tenella and Eimeria maxima are important pathogens causing intracellular protozoa infections in laboratory avian animals and are known to affect experimental results obtained from contaminated animals. This study aimed to find a fast, sensitive, and efficient protocol for the molecular identification of E. tenella and E. maxima in experimental samples using chickens as laboratory avian animals. DNA was extracted from fecal samples collected from chickens and polymerase chain reaction (PCR) analysis was employed to detect E. tenella and E. maxima from the extracted DNA. The target nucleic acid fragments were specifically amplified by PCR. Feces secreting E. tenella and E. maxima were detected by a positive PCR reaction. In this study, we were able to successfully detect E. tenella and E. maxima using the molecular diagnostic method of PCR. As such, we recommended PCR for monitoring E. tenella and E. maxima in laboratory avian facilities.

  14. 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

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

    Directory of Open Access Journals (Sweden)

    Zhang Zhenchao

    2017-01-01

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

  16. Molecular Characterization and Analysis of a Novel Protein Disulfide Isomerase-Like Protein of Eimeria tenella

    OpenAIRE

    Han, Hongyu; Dong, Hui; Zhu, Shunhai; Zhao, Qiping; Jiang, Lianlian; Wang, Yange; Li, Liujia; Wu, Youlin; Huang, Bing

    2014-01-01

    Protein disulfide isomerase (PDI) and PDI-like proteins are members of the thioredoxin superfamily. They contain thioredoxin-like domains and catalyze the physiological oxidation, reduction and isomerization of protein disulfide bonds, which are involved in cell function and development in prokaryotes and eukaryotes. In this study, EtPDIL, a novel PDI-like gene of Eimeria tenella, was cloned using rapid amplification of cDNA ends (RACE) according to the expressed sequence tag (EST). The EtPDI...

  17. Anticoccidial activity of hydrochloric acid (HCl against Eimeria tenella in broiler chickens

    Directory of Open Access Journals (Sweden)

    Rao Z Abbas

    2011-05-01

    Full Text Available The present study was planned to evaluate the anticoccidial activity of the different concentrations of the HCl against Eimeria tenella infection in broiler chickens in comparison with the amprolium anticoccidial. For this purpose, a total of 198 chicks were placed 11 per pen with three pens per treatment. The different concentrations of HCl (1000ppm, 2000ppm and 3000ppm and amproilum (at the dose rate of 125ppm were given to the experimental groups in drinking water from 10 to 19th days of age. One group was kept as infected non medicated control and one as non infected non medicated control. At the 12th day of age, all the groups were inoculated orally with 75,000 sporulated oocysts except non infected non medicated control. Anticoccidial activity was evaluated on the basis of performance (weight gain, feed conversion ratio and pathogenic (oocyst score, lesion score and mortality %age parameters. Among HCl medicated groups, the maximum anticoccidial effect was seen in the group medicated with 1000ppm HCl followed by 2000ppm and 3000ppm HCl medicated groups. Amprolium and 1000ppm HCl were almost equivalent in suppressing the negative performance and pathogenic effects associated with coccidiosis (Eimeria tenella challenge. In summary, the lower doses of HCl have the potential to be used as alternative to chemotherapeutic drugs for Eimeria tenella control. It is therefore suggested that further studies should be carried out to determine the possible minimum safe levels of HCl with least toxic effects to be used as anticoccidial.

  18. An Eimeria vaccine candidate based on Eimeria tenella immune mapped protein 1 and the TLR-5 agonist Salmonella typhimurium FliC flagellin

    Energy Technology Data Exchange (ETDEWEB)

    Yin, Guangwen; Qin, Mei [National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China); Liu, Xianyong [National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China); Key Laboratory of Zoonosis, China Ministry of Agriculture and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China); Suo, Jingxia; Tang, Xinming; Tao, Geru [National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China); Han, Qian [Department of Biochemistry, Virginia Tech, Blacksburg, VA 24061 (United States); Suo, Xun [National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China); Key Laboratory of Zoonosis, China Ministry of Agriculture and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China); Wu, Wenxue, E-mail: labboard@126.com [National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China); Key Laboratory of Zoonosis, China Ministry of Agriculture and College of Veterinary Medicine, China Agricultural University, Beijing 100193 (China)

    2013-10-25

    Highlights: •We found a new protective protein – (IMPI) in Eimeria tenella. •EtIMP1-flagellin fusion protein is an effective immunogen against Eimeria infection. •Flagellin can be as an apicomplexan parasite vaccine adjuvant in chickens. -- Abstract: Immune mapped protein-1 (IMP1) is a new protective protein in apicomplexan parasites, and exits in Eimeria tenella. But its structure and immunogenicity in E. tenella are still unknown. In this study, IMPI in E. tenella was predicted to be a membrane protein. To evaluate immunogenicity of IMPI in E. tenella, a chimeric subunit vaccine consisting of E. tenella IMP1 (EtIMP1) and a molecular adjuvant (a truncated flagellin, FliC) was constructed and over-expressed in Escherichia coli and its efficacy against E. tenella infection was evaluated. Three-week-old AA broiler chickens were vaccinated with the recombinant EtIMP1-truncated FliC without adjuvant or EtIMP1 with Freund’s Complete Adjuvant. Immunization of chickens with the recombinant EtIMP1-truncated FliC fusion protein resulted in stronger cellular immune responses than immunization with only recombinant EtIMP1 with adjuvant. The clinical effect of the EtIMP1-truncated FliC without adjuvant was also greater than that of the EtIMP1 with adjuvant, which was evidenced by the differences between the two groups in body weight gain, oocyst output and caecal lesions of E. tenella-challenged chickens. The results suggested that the EtIMP1-flagellin fusion protein can be used as an effective immunogen in the development of subunit vaccines against Eimeria infection. This is the first demonstration of antigen-specific protective immunity against avian coccidiosis using a recombinant flagellin as an apicomplexan parasite vaccine adjuvant in chickens.

  19. An Eimeria vaccine candidate based on Eimeria tenella immune mapped protein 1 and the TLR-5 agonist Salmonella typhimurium FliC flagellin

    International Nuclear Information System (INIS)

    Yin, Guangwen; Qin, Mei; Liu, Xianyong; Suo, Jingxia; Tang, Xinming; Tao, Geru; Han, Qian; Suo, Xun; Wu, Wenxue

    2013-01-01

    Highlights: •We found a new protective protein – (IMPI) in Eimeria tenella. •EtIMP1-flagellin fusion protein is an effective immunogen against Eimeria infection. •Flagellin can be as an apicomplexan parasite vaccine adjuvant in chickens. -- Abstract: Immune mapped protein-1 (IMP1) is a new protective protein in apicomplexan parasites, and exits in Eimeria tenella. But its structure and immunogenicity in E. tenella are still unknown. In this study, IMPI in E. tenella was predicted to be a membrane protein. To evaluate immunogenicity of IMPI in E. tenella, a chimeric subunit vaccine consisting of E. tenella IMP1 (EtIMP1) and a molecular adjuvant (a truncated flagellin, FliC) was constructed and over-expressed in Escherichia coli and its efficacy against E. tenella infection was evaluated. Three-week-old AA broiler chickens were vaccinated with the recombinant EtIMP1-truncated FliC without adjuvant or EtIMP1 with Freund’s Complete Adjuvant. Immunization of chickens with the recombinant EtIMP1-truncated FliC fusion protein resulted in stronger cellular immune responses than immunization with only recombinant EtIMP1 with adjuvant. The clinical effect of the EtIMP1-truncated FliC without adjuvant was also greater than that of the EtIMP1 with adjuvant, which was evidenced by the differences between the two groups in body weight gain, oocyst output and caecal lesions of E. tenella-challenged chickens. The results suggested that the EtIMP1-flagellin fusion protein can be used as an effective immunogen in the development of subunit vaccines against Eimeria infection. This is the first demonstration of antigen-specific protective immunity against avian coccidiosis using a recombinant flagellin as an apicomplexan parasite vaccine adjuvant in chickens

  20. Immunogenic Eimeria tenella glycosylphosphatidylinositol-anchored surface antigens (SAGs induce inflammatory responses in avian macrophages.

    Directory of Open Access Journals (Sweden)

    Yock-Ping Chow

    Full Text Available At least 19 glycosylphosphatidylinositol (GPI-anchored surface antigens (SAGs are expressed specifically by second-generation merozoites of Eimeria tenella, but the ability of these proteins to stimulate immune responses in the chicken is unknown.Ten SAGs, belonging to two previously defined multigene families (A and B, were expressed as soluble recombinant (r fusion proteins in E. coli. Chicken macrophages were treated with purified rSAGs and changes in macrophage nitrite production, and in mRNA expression profiles of inducible nitric oxide synthase (iNOS and of a panel of cytokines were measured. Treatment with rSAGs 4, 5, and 12 induced high levels of macrophage nitric oxide production and IL-1β mRNA transcription that may contribute to the inflammatory response observed during E. tenella infection. Concomitantly, treatment with rSAGs 4, 5 and 12 suppressed the expression of IL-12 and IFN-γ and elevated that of IL-10, suggesting that during infection these molecules may specifically impair the development of cellular mediated immunity.In summary, some E. tenella SAGs appear to differentially modulate chicken innate and humoral immune responses and those derived from multigene family A (especially rSAG 12 may be more strongly linked with E. tenella pathogenicity associated with the endogenous second generation stages.

  1. Shedding of the immunodominant P20 surface antigen of Eimeria bovis sporozoites.

    OpenAIRE

    Speer, C A; Whitmire, W M

    1989-01-01

    P20 is an immunodominant surface antigen of Eimeria bovis sporozoites. As parasites underwent merogony within cultured bovine monocytes and Madin-Darby bovine kidney (MDBK) cells, P20 appeared to be shed gradually by meronts and was absent in type 1 and 2 first-generation merozoites. Meronts of E. bovis appeared to shed P20 into the parasitophorous vacuole of bovine monocytes, whereas MDBK cells evidently released P20 into the culture medium or destroyed its antigenic determinant.

  2. Use of Eimeria tenella radiovaccine for immunoprophylaxis in fowl against coccidiosis

    International Nuclear Information System (INIS)

    Jungmann, R.; Mielke, D.

    1989-01-01

    Studies were conducted into oral immunization of broilers, using a widely standardized coccidiosis vaccine attenuated by irradiation with 250 Gy (Eimeria tenella radiovaccine). The method was applied to 2,567 animals kept in floor pens and to another 21,620 animals under production conditions. Profound immunity was thus built up to last from three weeks after vaccination to slaughter maturity. Immunized chickens challenged with E. tenella develope no clinical infection, whereas the non-immunized ones showed an intensive haemorrhagic enteritis with 88% of lethality. The total output of coccidiaoocysts of immunized chickens decreased to more than 95% compared to the non-immunized ones. These results were reproducible in six different experimental groups. (author)

  3. Study on the immunogenic ability of Eimeria tenella oocytes following treatment with gamma rays

    International Nuclear Information System (INIS)

    Penev, P.; Stafanova, M.

    1975-01-01

    Studied was the immunizing capacity of Eimeria tenella oocytes, treated with gamma rays at the rate of 6000 R, in 10- and 20-day-old chickens. The oocytes sporulated after treatment. Applied at the rate of 50,000 R they showed lower virulence and were capable of inducing resistance to reinfection with non-irradiated oocytes at rates that were three times as much. Following reinfection some birds manifested subclinical coccidiosis but survived. This showed that the immunization with oocytes that had been irradiated with 6,000 R had its peculiar aspects. (author)

  4. Characterization of the Eimeria maxima sporozoite surface protein IMP1

    Science.gov (United States)

    The purpose of this study was to characterize Eimeria maxima immunoprotective protein IMP1 that is hypothesized to play a role in eliciting protective immunity against E. maxima infection in chickens. RT-PCR analysis of RNA from unsporulated and sporulating E. maxima oocysts revealed highest transc...

  5. Expression and immunological characterisation of Eimeria tenella glycosylphosphatidylinositol-anchored surface antigen-5

    Science.gov (United States)

    Ho, Sue-Kim; Nathan, Sheila; Wan, Kiew-Lian

    2016-11-01

    Eimeria tenella is the most pathogenic of the Eimeria species that infect chickens and causes huge economic losses to the poultry industry. The glycosylphosphatidylinositol-anchored surface antigen-5 (SAG5) found on the surface of the parasite has been shown to activate the chicken's immune system. In this study, recombinant SAG5 was expressed, purified and used to investigate the immune-inducing characteristics of the molecule. Chickens were immunized with purified recombinant SAG5 and sera were subjected to Enzyme-linked Immunosorbant Assay (ELISA). Results indicated that specific antibodies against rSAG5 were produced, with IgG detected at a higher level compared to IgA and IgM. Information on the immunological responses elicited by SAG5 provides essential knowledge that will contribute towards the effort to develop more effective strategies against coccidiosis.

  6. Irradiated Eimeria brunetti, E. necatrix and E. tenella in the simultaneous immunization of chickens

    International Nuclear Information System (INIS)

    Sokolic, A.; Movsesijan, M.; Tanielian, Z.; Abu Ali, N.

    1976-01-01

    Chickens three weeks of age were immunized simultaneously against E. necatrix, E. tenella and E. brunetti receiving a single oral dose of oocysts of these Eimeria spp. irradiated at 10k rads with gamma rays delivered from a 60 Co-source. Two weeks later immunized chickens and their corresponding untreated controls were challenged with infective oocysts of the same three protozoan species. The results obtained have shown that all immunized chickens survived a heavy challenge which killed 70% of the corresponding control chickens. The results and their possible practical implication are discussed. Eimeria spp. selected for these studies were of great epidemiological and economic importance in the area of Lebanon with the most intensive poultry production. (author)

  7. Insights into the genome structure and copy-number variation of Eimeria tenella

    Directory of Open Access Journals (Sweden)

    Lim Lik-Sin

    2012-08-01

    Full Text Available Abstract Background Eimeria is a genus of parasites in the same phylum (Apicomplexa as human parasites such as Toxoplasma, Cryptosporidium and the malaria parasite Plasmodium. As an apicomplexan whose life-cycle involves a single host, Eimeria is a convenient model for understanding this group of organisms. Although the genomes of the Apicomplexa are diverse, that of Eimeria is unique in being composed of large alternating blocks of sequence with very different characteristics - an arrangement seen in no other organism. This arrangement has impeded efforts to fully sequence the genome of Eimeria, which remains the last of the major apicomplexans to be fully analyzed. In order to increase the value of the genome sequence data and aid in the effort to gain a better understanding of the Eimeria tenella genome, we constructed a whole genome map for the parasite. Results A total of 1245 contigs representing 70.0% of the whole genome assembly sequences (Wellcome Trust Sanger Institute were selected and subjected to marker selection. Subsequently, 2482 HAPPY markers were developed and typed. Of these, 795 were considered as usable markers, and utilized in the construction of a HAPPY map. Markers developed from chromosomally-assigned genes were then integrated into the HAPPY map and this aided the assignment of a number of linkage groups to their respective chromosomes. BAC-end sequences and contigs from whole genome sequencing were also integrated to improve and validate the HAPPY map. This resulted in an integrated HAPPY map consisting of 60 linkage groups that covers approximately half of the estimated 60 Mb genome. Further analysis suggests that the segmental organization first seen in Chromosome 1 is present throughout the genome, with repeat-poor (P regions alternating with repeat-rich (R regions. Evidence of copy-number variation between strains was also uncovered. Conclusions This paper describes the application of a whole genome mapping

  8. Effects of Mushroom and Herb Polysaccharides on Cellular and Humoral Immune Responses of Eimeria tenella-Infected Chickens

    NARCIS (Netherlands)

    Guo, F.C.; Kwakkel, R.P.; Williams, B.A.; Parmentier, H.K.; Li, W.K.; Yang, Z.Q.; Verstegen, M.W.A.

    2004-01-01

    We investigated the effects of polysaccharide extracts from 2 mushrooms, Lentinus edodes (LenE) and Tremella fuciformis (TreE), and an herb, Astragalus membranaceus (AstE), on cellular and humoral immune responses of Eimeria tenella-infected chickens. A total of 150 broiler chicks were assigned to 5

  9. Coccidiosis Immunization: Effects of Mushroom and Herb Polysaccharides on Immune Responses of Chickens Infected with Eimeria Tenella

    NARCIS (Netherlands)

    Guo, F.C.; Kwakkel, R.P.; Williams, B.A.; Suo, X.; Li, W.K.; Verstegen, M.W.A.

    2005-01-01

    An experiment was conducted to investigate the effects of polysaccharide extracts (E) of two mushrooms, Lentinus edodes (LenE) and Tremella fuciformis (TreE), and an herb, Astragalus membranaceus (AstE), on the immune responses of chickens infected with Eimeria tenella. A total of 180 broiler

  10. Anti-coccidial activity of the ethanol extract of Tribulus terrestris fruits on Eimeria tenella

    Science.gov (United States)

    Hong, Sunhwa; Moon, Mi-Na; Im, Eun-Kyung; Won, Jum-Soon; Yoo, Ji-Hyun

    2018-01-01

    Anti-coccidial effects of the fruits of Tribulus terrestris (Tribuli fructus) ethanol extract (TTE) were studied with animal experiment following per oral administration with Eimeria (E.) tenella. This experiment was performed on the 3-day-old chicks (n=30). The animals were divided with 3 groups; TFE 15mg per animal+infected (n=10), TTE untreated+infected (n=10) and non-infected control (n=10). Animals were administrated with or without TTE during 1 week, and then inoculated with E. tenella. The anti-coccidial activity were evaluated with oocysts shedding numbers in stools, body weights changes and food intake changes. The TTE-inoclated animals revealed significantly decreased stool oocysts numbers (P<0.05) when compared to the TTE untreated animals. Also, TTE-treated animals showed more increased body weight gains (P<0.05) than the TTE untreated animals. These results demonstrate that TTE produce anticoccidial activities against E. tenella. TTE could be a promising treatment for the coccidiosis. PMID:29628976

  11. Anti-coccidial activity of the ethanol extract of Tribulus terrestris fruits on Eimeria tenella.

    Science.gov (United States)

    Hong, Sunhwa; Moon, Mi-Na; Im, Eun-Kyung; Won, Jum-Soon; Yoo, Ji-Hyun; Kim, Okjin

    2018-03-01

    Anti-coccidial effects of the fruits of Tribulus terrestris (Tribuli fructus) ethanol extract (TTE) were studied with animal experiment following per oral administration with Eimeria ( E .) tenella . This experiment was performed on the 3-day-old chicks (n=30). The animals were divided with 3 groups; TFE 15mg per animal+infected (n=10), TTE untreated+infected (n=10) and non-infected control (n=10). Animals were administrated with or without TTE during 1 week, and then inoculated with E. tenella . The anti-coccidial activity were evaluated with oocysts shedding numbers in stools, body weights changes and food intake changes. The TTE-inoclated animals revealed significantly decreased stool oocysts numbers ( P <0.05) when compared to the TTE untreated animals. Also, TTE-treated animals showed more increased body weight gains ( P <0.05) than the TTE untreated animals. These results demonstrate that TTE produce anticoccidial activities against E. tenella . TTE could be a promising treatment for the coccidiosis.

  12. Protection Efficacy of Multivalent Egg Yolk Immunoglobulin against Eimeria Tenella Infection in Chickens

    Directory of Open Access Journals (Sweden)

    JJ Xu

    2013-09-01

    Full Text Available Background: To control avian coccidiosis with drug-independent strategy effec­tively and safely, multivalent hyperimmune egg yolk immunoglobulin (IgY was prepared and its ability to protect against Eimeria tenella infection was evaluated.Methods: Hens were orally immunized with live oocysts of 5 species of Eimeria for six times, antibody titers in serum and yolk were monitored by indirect enzyme-linked immunosorbent assay. The specific IgY was isolated, purified and lyophi­lized. IgY powder was orally administrated as dietary supplement in newly hatched chicks at various dosages. Birds were orally challenged with 10000 sporulated oo­cysts of E. tenella at 10 days of age, weighed and killed at 8 days post challenge, and the protective effect was assessed.Results: The averge yeid of IgY was 9.2 mg/ml yolk, the antibody titer of IgY reached to 1:163840 per mg with the purity up to 98%. Chickens fed IgY resulted in reduced mortality, increased body weight gain (BWG, reduced oocyst shedding, reduced caecal lesion score and increased anti-coccidial index. In terms of BWG and caecal lesion, IgY significantly enhanced the resistance of bird at ≥ 0.05% of IgY in the diet when compared with the challenged control group (P0.05.Conclusion: Supplementing newly hatched chicks with Eimeria-specific IgY represents a promising strategy to prevent avian coccidiosis.

  13. Monitoring of local CD8 β-expressing cell populations during Eimeria tenella infection of naïve and immune chickens

    DEFF Research Database (Denmark)

    Wattrang, Eva; Thebo, Per; Lunden, Anna

    2016-01-01

    The purpose of this study was to monitor abundance and activation of local CD8β-expressing T-cell populations during Eimeria tenella infections of naïve chickens and chickens immune by previous infections. Chickens were infected with E. tenella up to three times. Caecal T-cell receptor (TCR) γ...

  14. Studies on the effects of whole-body gamma irradiation on chickens infected with Eimeria tenella

    International Nuclear Information System (INIS)

    Merritt, S.V.

    1974-01-01

    Whole-body exposure of one- and three-week-old White Leghorn cockerels to 600 R gamma radiation (Cesium-137) 24 hours before oral inoculation with 500, 2500, 5000, or 50,000 Eimeria tenella oocysts produced a pattern of mortality differing markedly from nonirradiated, infected (NRI) control birds. When oocyst dosage was held constant (2500) and radiation exposure increased (250, 450, 600, 800, or 1000 R) a gradual increase in mortality rate with higher radiation dosages was observed among both one- and three-week-old birds. Birds irradiated 24 hours or more before inoculation were less able to survive infection than were those irradiated one hour before and one, two, three, or four days after inoculation. (U.S.)

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

    Science.gov (United States)

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

    2007-01-01

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

  16. Location of RAD51-like protein during meiotic prophase in Eimeria tenella.

    Science.gov (United States)

    Del Cacho, Emilio; Gallego, Margarita; Pagés, Marc; Barbero, José Luís; Monteagudo, Luís; Sánchez-Acedo, Caridad

    2011-05-31

    This study focuses on reporting events in Eimeria tenella oocysts from early to late prophase I in terms of RAD51 protein in association with the synaptonemal complex formed between homologous chromosomes. The aim of the study was the sequential localization of RAD51 protein, which is involved in the repair of double-strand breaks (DSBs) on the eimerian chromosomes as they synapse and desynapse. Structural Maintenance of Chromosome protein SMC3, which plays a role in synaptonemal complex formation, was labeled to identify initiation and progress of chromosome synapsis and desynapsis in parallel with the appearance and disappearance of RAD51 foci. Antibodies directed against RAD51 and cohesin subunit SMC3 proteins were labeled with either fluorescence or colloidal gold to visualize RAD51 protein foci and synaptonemal complexes. RAD51 protein localization during prophase I was studied on meiotic chromosomes spreads obtained from oocysts at different points in time after the start of sporulation. The present findings showed that foci detected with the antibody directed against RAD51 protein first appeared at the pre-leptotene stage before homologous chromosomes began pairing. Subsequently, the foci were detected in association with the lateral elements at the precise sites where synapsis were in progress. These findings lead us to suggest that in E. tenella, homologous chromosome pairing was a DSB-dependent mechanism and reinforced the participation of RAD51 protein in meiotic homology search, alignment and pairing of chromosomes. Copyright © 2010 Elsevier B.V. All rights reserved.

  17. Evaluation of plasma chemistry and haematological studies on chickens infected with Eimeria tenella and E acervulina.

    Science.gov (United States)

    Fukata, T; Komba, Y; Sasai, K; Baba, E; Arakawa, A

    1997-07-12

    Plasma chemistry and haematological studies were conducted on chickens with coccidiosis. Male White Leghorn chickens, of two weeks old, were inoculated with 5 x 10(4) Eimeria tenella sporulated oocysts or with 1 x 10(6) E acervulina sporulated oocysts. Blood samples were taken four, seven and 11 days after inoculation. A wet chemistry system was applied to measure the plasma activities of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma glutamyltransferase, creatine kinase, amylase and lactate dehydrogenase and the concentrations of creatine, total bilirubin, urate, total cholesterol, total protein, albumin, glucose and triglycerides. A dry chemistry system was applied to measure sodium, potassium, chloride and calcium. The number of red blood cells and packed cell volume were determined by a micro cell counter and blood pH was measured with a blood gas analyser. The erythrocyte count, packed cell volume, sodium and chloride levels in the chickens infected with E tenella were significantly (P < 0.05) lower than those of the uninfected controls. The significant decrease in blood pH of the chickens infected with E acervulina suggests malabsorption associated with duodenal lesions induced by the infection.

  18. Identification of sporozoite surface proteins and antigens of Eimeria nieschulzi (Apicomplexa)

    International Nuclear Information System (INIS)

    Tilley, M.; Upton, S.J.

    1990-01-01

    Sodium dodecyl sulfate polyacrylamide gel electrophoresis, immunoblotting, lectin binding, and 125 I surface labeling of sporozoites were used to probe sporozoites of the rat coccidian, Eimeria nieschulzi. Analysis of silver stained gels revealed greater than 50 bands. Surface iodination revealed about 14 well labeled, and about 10 weakly labeled but potential, surface proteins. The most heavily labeled surface proteins had molecular masses of 60, 53-54, 45, 28, 23-24, 17, 15, 14, 13, and 12 kD. Following electrophoresis and Western blotting, 2 of the 12 125I labeled lectin probes bound to two bands on the blots, which collectively indicated that two bands were glycosylated. Concanavalin A (ConA) specifically recognized a band at 53 kD, which may represent a surface glycoprotein, and a lectin derived from Osage orange (MPA) bound to a single band at 82-88 kD, that may also be a surface molecule. Immunoblotting using sera collected from rats inoculated orally with oocysts, as well as sera from mice hyperimmunized with sporozoites, revealed that many surface molecules appear to be immunogenic

  19. Effects of dietary supplementation with phytonutrients on vaccine-stimulated immunity against infection with Eimeria tenella.

    Science.gov (United States)

    Lee, Sung Hyen; Lillehoj, Hyun S; Jang, Seung I; Lee, Kyung Woo; Bravo, David; Lillehoj, Erik P

    2011-09-27

    Two phytonutrient mixtures, VAC (carvacrol, cinnamaldehyde, and Capsicum oleoresin), and MC (Capsicum oleoresin and turmeric oleoresin), were evaluated for their effects on chicken immune responses following immunization with an Eimeria profilin protein. Chickens were fed with a non-supplemented diet, or with VAC- or MC-supplemented diets, immunized with profilin, and orally challenged with virulent oocysts of Eimeria tenella. Immunity against infection was evaluated by body weight, fecal oocyst shedding, profilin antibody levels, lymphocyte recall responses, cytokine expression, and lymphocyte subpopulations. Following immunization and infection, chickens fed the VAC- or MC-supplemented diets showed increased body weights, greater profilin antibody levels, and/or greater lymphocyte proliferation compared with non-supplemented controls. Prior to Eimeria infection, immunized chickens on the MC-supplemented diet showed reduced IFN-γ and IL-6 levels, but increased expression of TNFSF15, compared with non-supplemented controls. Post-infection levels of IFN-γ and IL-6 were increased, while IL-17F transcripts were decreased, with MC-supplementation. For VAC-supplemented diets, decreased IL-17F and TNFSF15 levels were observed only in infected chickens. Finally, immunized chickens fed the MC-supplemented diet exhibited increased MHC class II(+), CD4(+), CD8(+), TCR1+, or TCR2(+) T cells compared with nonsupplemented controls. Animals on the VAC-containing diet only displayed an increase in K1(+) macrophages. In conclusion, dietary supplementation with VAC or MC alters immune parameters following recombinant protein vaccination against avian coccidiosis. Published by Elsevier B.V.

  20. Efficacy of chimeric DNA vaccines encoding Eimeria tenella 5401 and chicken IFN-γ or IL-2 against coccidiosis in chickens.

    Science.gov (United States)

    Song, Xiaokai; Huang, Xinmei; Yan, Ruofeng; Xu, Lixin; Li, Xiangrui

    2015-09-01

    Chimeric DNA vaccines encoding Eimeria tenella (E. tenella) surface antigen 5401 were constructed and their efficacies against E. tenella challenge were studied. The open reading frame (ORF) of 5401 was cloned into the prokaryotic expression vector pGEX-4T2 to express the recombinant protein and the expressed recombinant protein was identified by Western blot. The ORF of 5401 and chicken cytokine gene IFN-γ or IL-2 were cloned into the eukaryotic expression vector pVAX1 consecutively to construct DNA vaccines pVAX-5401-IFN-γ, pVAX-5401-IL-2 and pVAX-5401. The expression of aim genes in vivo was detected by reverse transcription-polymerase chain reaction and Western blot. Fourteen-day-old chickens were inoculated twice at an interval of 7 days with 100 µg of plasmids pVAX-5401, pVAX-5401-IFN-γ and pVAX-5401-IL-2 or 200 µg of recombinant 5401 protein by leg intramuscular injection, respectively. Seven days after the second inoculation, all chickens except the unchallenged control group were challenged orally with 5 × 10(4) sporulated oocysts of E. tenella. Seven days after challenge, all chickens were weighted and slaughtered to determine the effects of immunization. The results showed the recombinant protein was about 90 kDa and reacted with antiserum against soluble sporozoites. The animal experiment showed that all the DNA vaccines pVAX-5401, pVAX-5401-IFN-γ or pVAX-5401-IL-2 and the recombinant 5401 protein could obviously alleviate body weight loss and cecal lesions as compared with non-vaccinated challenged control and empty vector pVAX1control. Furthermore, pVAX-5401-IFN-γ or pVAX-5401-IL-2 induced anti-coccidial index (ACI) of 180.01 or 177.24 which were significantly higher than that of pVAX-5401. The results suggested that 5401 was an effective candidate antigen for vaccine. This finding also suggested that chicken IFN-γ or IL-2 could effectively improve the efficacies of DNA vaccines against avian coccidiosis. Copyright © 2015 Elsevier

  1. 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. Copyright © 2015 Elsevier B.V. All rights

  2. The response of red ginger (Zinggiber officinalle var rubra) with various processing in broilers were infected by Eimeria tenella

    Science.gov (United States)

    Nasution, E. Z. J.; Tafsin, M.; Hanafi, N. D.

    2018-02-01

    Red ginger contains high antioxidants and have anti -inflamatory properties. Ginger also has the ability to treat kimiatif, antiemetic, antinausea, and antiparasitik. The aim of this experiment was identified the response of red ginger in broilers were infected by Eimeria tenella. This research used Completely Randomized Design (CRD) with 5 treatments and 4 replications. Eimeria tenella were infected by 10.000 oocysts / head and red ginger solution were aplicated with 1% concentration. The treatments consist of KP (positive control), KO (coccidiostat), K1 (red ginger powder), K2 (extracted red ginger by ethanol) and K3 (extracted red ginger by water) . The results showed that the treatment of red ginger was significant effect (PEimeria tenella. The comparison between extracted red ginger by ethanol is better than by water or in powder form to decreased. The utilization of red ginger showed the percentage of heterophile and eosinophile close to normal when compared with positive control. Assesment of caecum lesion score was not significant (P>0.05) different effect between all the treatments. It is concluded that the treatment by red ginger better than coccidiostat and positive control.

  3. 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.

  4. Vaccination with Eimeria tenella Elongation Factor-1alpha Recombinant Protein Induces protective Immunity against E. tenella and E. maxima infections

    Science.gov (United States)

    Avian coccidiosis is caused by multiple species of the apicomplexan protozoan, Eimeria, and is one of the most economically devastating enteric diseases for the poultry industry worldwide. Host immunity to Eimeria infection, however, is relatively species-specific. The ability to immunize chickens a...

  5. Comparative Microarray Analysis of Intestinal Lymphocytes following Eimeria acervulina, E. maxima, or E. tenella Infection in the Chicken

    Science.gov (United States)

    Kim, Duk Kyung; Lillehoj, Hyun; Min, Wongi; Kim, Chul Hong; Park, Myeong Seon; Hong, Yeong Ho; Lillehoj, Erik P.

    2011-01-01

    Relative expression levels of immune- and non-immune-related mRNAs in chicken intestinal intraepithelial lymphocytes experimentally infected with Eimeria acervulina, E. maxima, or E. tenella were measured using a 10K cDNA microarray. Based on a cutoff of >2.0-fold differential expression compared with uninfected controls, relatively equal numbers of transcripts were altered by the three Eimeria infections at 1, 2, and 3 days post-primary infection. By contrast, E. tenella elicited the greatest number of altered transcripts at 4, 5, and 6 days post-primary infection, and at all time points following secondary infection. When analyzed on the basis of up- or down-regulated transcript levels over the entire 6 day infection periods, approximately equal numbers of up-regulated transcripts were detected following E. tenella primary (1,469) and secondary (1,459) infections, with a greater number of down-regulated mRNAs following secondary (1,063) vs. primary (890) infection. On the contrary, relatively few mRNA were modulated following primary infection with E. acervulina (35 up, 160 down) or E. maxima (65 up, 148 down) compared with secondary infection (E. acervulina, 1,142 up, 1,289 down; E. maxima, 368 up, 1,349 down). With all three coccidia, biological pathway analysis identified the altered transcripts as belonging to the categories of “Disease and Disorder” and “Physiological System Development and Function”. Sixteen intracellular signaling pathways were identified from the differentially expressed transcripts following Eimeria infection, with the greatest significance observed following E. acervulina infection. Taken together, this new information will expand our understanding of host-pathogen interactions in avian coccidiosis and contribute to the development of novel disease control strategies. PMID:22140460

  6. Ameliorative effect of mycofix plus 3.0 in reducing intensity of Eimeria tenella infection during aflatoxicosis in broiler chicks

    Directory of Open Access Journals (Sweden)

    A. M. Shareef

    2008-01-01

    Full Text Available One hundred and sixty male broiler chicks were fed at one day of age aflatoxin (AF at a rate of 3.5 mg/kg alone, or with groups injected with Eimeria tenella sporulated oocysts (40000 at 14 days of age. Adsorbent (Mycofix® plus 3.0 was incorporated at a rate of 0.25% in the above mentioned groups from one day of age till the end of the experiment. The study was conducted to reveal the effect of a aforementioned different diets and treatments on live body weight, feed consumption, feed conversion ratio, blood parameters (total red blood cells, hemoglobin, packed cell volume, biochemical profile of serum (alkaline phosphatase and β-carotin, liver weights, bursal and thymus indexes, caecal lesion scores and mortalities. The results indicated that AF was responsible for a significant (P<0.05 reduction. in body weigh gain (BWG, feed consumption, and an increase in feed conversion ratio. Afllatoxin was also responsible for reduction in blood parameters, β-Carotin, bursal and thymus indexes. While relative liver weight and alkaline phosphatase level were significantly (P<0.05 increased. Groups that fed AF at a rate of 3.5 mg/kg feed and exposed to sporulated oocysts of Eimeria tenella show a high significant (P<0.05 reduction in BWG, feed consumption and an increase in feed conversion ratio. Aflatoxin was also responsible for significant blood parameter, β-carotin, and also a significant (P<0.05 increase in caecal lesion scores, mortality, alkaline phosphatase level and relative liver weight, while they showed significant (P<0.05 decrease in bursal and thymus indexes in comparison with injected groups with E-tenella sporulated oocysts alone. The study approved that the groups maintained on mycofix plus 3.0 (0.25% and contaminated with aflatoxin 3.5 mg/kg, revealed a positive noticeable effects in amelioration on BWG, feed consumption and feed conversion, blood parameter, β-carotin, alkaline phosphatase level, relative liver weight, bursal and

  7. EVALUATION OF THE ANTIGENICITY AND IMMUNOGENICITY OF Eimeria tenella BY REPRODUCTIVE INDEX AND HISTOPATHOLOGICAL CHANGES OF CECAL COCCIDIOSIS VIRULENT LIVE VACCINE IN BROILER CHICKENS.

    Science.gov (United States)

    Suprihati, Endang; Yunus, Muchammad

    2018-01-01

    The development of vaccine to control coccidiosis caused by Eimeria tenella ( E. tenella ) in chickens is intensifying because of the increasing threat of drug resistance to anticoccidial agents. It is important, therefore, to develop a reliable standard method for the assessment of vaccine afficacy particularly antigenicity and immunogenicity become crucial. Evaluation of E. tenella antigenicity and immunogenicity to some low doses can be reflected by reproductive index and histopathological changes. The complete random design of research was used in this study. Sixty of two weeks old broilers were divided into four groups and each group composed 15 replications. The group 1 was chicken group without virulent E. tenella oocyst inoculation. The group 2, 3 and group 4 were chicken group inoculated with virulent E. tenella oocyst at doses of 1.0 x 10 2 , 2.0 x 10 2 , 3.0 x 10 2 , respectively. Then all chicken groups were challenged with E. tenella oocyst at doses of 1.0 x 10 3 . Observation of research that represented antigenicity and immunogenicity was clinical sign, reproductive index, histopathological changes. On virulent E. tenella inoculation step, some clinical signs such as appetite, weakness, and diarrhea were very slight on all chicken groups. While on challenge test step, there were no clinical signs of all chicken groups except the group 1. For the reproductive index of virulent E. tenella inoculation step, there were no significantly differences in all chicken groups except the group 1. As reproductive index, the same result pattern was seen for histopathological changes. The low number virulent E. tenella had low reproductive index and few histopathological changes effect that represents a promising strategy to prevent cecal coccidiosis in chickens.

  8. Profil Saraf Nitrergik Sekum Ayam Pedaging yang Diinfeksi Eimeria tenella (THE PROFILE OF NEORAL NITRERGIC IN THE COECUM OF BROILER INFECTED WITH Emeria tenella

    Directory of Open Access Journals (Sweden)

    Amelia Hana

    2016-01-01

    Full Text Available Neurotransmitter found in the enteric nervous system that plays an important role in a variety ofenteropathies, including inflammatory bowel disease. Alteration of nitrergic neurons has been reported to bedependent on the manner by which inflammation is caused. This study was performed to determine theprofile of neural nitrergic with Nicotinamide Adenine Dinucleotide Phosphate-diaphorase (NADPH-dstaining in the cecum of broiler infected by Eimeria tenella (E. tenella. Thirty six health free of coccidiosisbroiler one-day old chickens (DOC used in this study. All of broiler adapted for 7 days with food and drinkgiven ad libitum then divided into three groups, each group consisted of 12 broilers. The 1st group wascontrol only given orally 1.0 ml of distilled water/ heads , while 2nd group was infected with a single dose of5x103 oocysts/ head orally, and 3rd group was infected with a single dose of 2x104 oocysts/ head orally. Onday 7 post infection, all of chickens were fasted for 12 hours then were euthanized and cecum was taken.Lesion score of cecum was assessed. Furthermore, tissue of the coecum was prepared for hisochemicalusing NADPH-d staining to determine morphology, and morhometric of nitrergic neurons. The resultshown that cecum lesion score of group I is 0, group II is +2, group III is +3. Neuron density in the cecum ofgroup I is 2794.96+4.65 neuron/cm2, group II is 4930.36+63.73 neuron/cm2 and group III is 7892.31+44.97 neuron/cm2. Neuron density of group II and III increased signifcantly (p<001 than group I. Nitrergic neurondesity of group III increased significantly (p<0.05 than that of group II. It was concluded that the infectionof E.tenella led to increase the number of neutrl mienteric nitrergic of the cecum. The higher lesion score ofcecum led to increase the number of mienteric nitrergic neuron.

  9. Host response to simultaneous infections with Eimeria acervulina, maxima and tenella: A cumulation of single responses

    NARCIS (Netherlands)

    Cornelissen, J.B.W.J.; Swinkels, W.J.C.; Boersma, W.A.; Rebel, J.M.J.

    2009-01-01

    It is well known that broilers may be infected by different Eimeria strains at the same time and that different species infect specific parts of the gut. Cell mediated responses play a major role in the immune response in broilers after infection with Eimeria species. The cell mediated responses

  10. Invasion of the intestinal tract by sporozoites of Eimeria coecicola and Eimeria intestinalis in naive and immune rabbits

    Czech Academy of Sciences Publication Activity Database

    Pakandl, Michal; Sewald, B.; Drouet-Viard, F.

    2006-01-01

    Roč. 98, č. 4 (2006), s. 310-316 ISSN 0932-0113 R&D Projects: GA ČR GA524/05/2328 Institutional research plan: CEZ:AV0Z60220518 Keywords : rabbit coccidia * sporozoites * intra- and extraintestinal migration Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 1.140, year: 2006

  11. The Induction of Protective Immunity against Experimental Eimeria tenella Infection using Serum Exosomes

    Science.gov (United States)

    Avian coccidiosis is caused by Eimeria, a unicellular, apicomplexan protist which primarily infects intestinal epithelia resulting in nutrition malabsorption and reduced growth of commercial poultry. Vaccination of chickens with exosomes isolated from antigen presenting cells and containing parasit...

  12. Effect of dietary vitamin E on Eimeria tenella-induced oxidative ...

    African Journals Online (AJOL)

    ramezanali

    2012-05-10

    May 10, 2012 ... Full Length Research Paper. Effect of dietary ... vitamin E (p<0.05). A significant rise of plasma and erythrocyte MDA was ... biological membranes, thereby making them stable. This prevents free ... Vegetable oil. 2.00. 2.00 ..... tenella: a review of the type strain selected for genome sequencing. Avian Pathol.

  13. Microsatellite mapping of QTLs affecting resistance to coccidiosis (Eimeria tenella) in a Fayoumi x White Leghorn cross.

    Science.gov (United States)

    Pinard-van der Laan, Marie-Hélène; Bed'hom, Bertrand; Coville, Jean-Luc; Pitel, Frédérique; Feve, Katia; Leroux, Sophie; Legros, Hélène; Thomas, Aurélie; Gourichon, David; Repérant, Jean-Michel; Rault, Paul

    2009-01-20

    Avian coccidiosis is a major parasitic disease of poultry, causing severe economical loss to poultry production by affecting growth and feed efficiency of infected birds. Current control strategies using mainly drugs and more recently vaccination are showing drawbacks and alternative strategies are needed. Using genetic resistance that would limit the negative and very costly effects of the disease would be highly relevant. The purpose of this work was to detect for the first time QTL for disease resistance traits to Eimeria tenella in chicken by performing a genome scan in an F2 cross issued from a resistant Fayoumi line and a susceptible Leghorn line. The QTL analysis detected 21 chromosome-wide significant QTL for the different traits related to disease resistance (body weight growth, plasma coloration, hematocrit, rectal temperature and lesion) on 6 chromosomes. Out of these, a genome-wide very significant QTL for body weight growth was found on GGA1, five genome-wide significant QTL for body weight growth, plasma coloration and hematocrit and one for plasma coloration were found on GGA1 and GGA6, respectively. Two genome-wide suggestive QTL for plasma coloration and rectal temperature were found on GGA1 and GGA2, respectively. Other chromosme-wide significant QTL were identified on GGA2, GGA3, GGA6, GGA15 and GGA23. Parent-of-origin effects were found for QTL for body weight growth and plasma coloration on GGA1 and GGA3. Several QTL for different resistance phenotypes were identified as co-localized on the same location. Using an F2 cross from resistant and susceptible chicken lines proved to be a successful strategy to identify QTL for different resistance traits to Eimeria tenella, opening the way for further gene identification and underlying mechanisms and hopefully possibilities for new breeding strategies for resistance to coccidiosis in the chicken. From the QTL regions identified, several candidate genes and relevant pathways linked to innate immune

  14. Microsatellite mapping of QTLs affecting resistance to coccidiosis (Eimeria tenella in a Fayoumi × White Leghorn cross

    Directory of Open Access Journals (Sweden)

    Gourichon David

    2009-01-01

    Full Text Available Abstract Background Avian coccidiosis is a major parasitic disease of poultry, causing severe economical loss to poultry production by affecting growth and feed efficiency of infected birds. Current control strategies using mainly drugs and more recently vaccination are showing drawbacks and alternative strategies are needed. Using genetic resistance that would limit the negative and very costly effects of the disease would be highly relevant. The purpose of this work was to detect for the first time QTL for disease resistance traits to Eimeria tenella in chicken by performing a genome scan in an F2 cross issued from a resistant Fayoumi line and a susceptible Leghorn line. Results The QTL analysis detected 21 chromosome-wide significant QTL for the different traits related to disease resistance (body weight growth, plasma coloration, hematocrit, rectal temperature and lesion on 6 chromosomes. Out of these, a genome-wide very significant QTL for body weight growth was found on GGA1, five genome-wide significant QTL for body weight growth, plasma coloration and hematocrit and one for plasma coloration were found on GGA1 and GGA6, respectively. Two genome-wide suggestive QTL for plasma coloration and rectal temperature were found on GGA1 and GGA2, respectively. Other chromosme-wide significant QTL were identified on GGA2, GGA3, GGA6, GGA15 and GGA23. Parent-of-origin effects were found for QTL for body weight growth and plasma coloration on GGA1 and GGA3. Several QTL for different resistance phenotypes were identified as co-localized on the same location. Conclusion Using an F2 cross from resistant and susceptible chicken lines proved to be a successful strategy to identify QTL for different resistance traits to Eimeria tenella, opening the way for further gene identification and underlying mechanisms and hopefully possibilities for new breeding strategies for resistance to coccidiosis in the chicken. From the QTL regions identified, several

  15. Proteomic analysis of protein interactions between Eimeria maxima sporozoites and chicken jejunal epithelial cells by shotgun LC-MS/MS.

    Science.gov (United States)

    Huang, Jingwei; Liu, Tingqi; Li, Ke; Song, Xiaokai; Yan, Ruofeng; Xu, Lixin; Li, Xiangrui

    2018-04-04

    Eimeria maxima initiates infection by invading the jejunal epithelial cells of chicken. However, the proteins involved in invasion remain unknown. The research of the molecules that participate in the interactions between E. maxima sporozoites and host target cells will fill a gap in our understanding of the invasion system of this parasitic pathogen. In the present study, chicken jejunal epithelial cells were isolated and cultured in vitro. Western blot was employed to analyze the soluble proteins of E. maxima sporozoites that bound to chicken jejunal epithelial cells. Co-immunoprecipitation (co-IP) assay was used to separate the E. maxima proteins that bound to chicken jejunal epithelial cells. Shotgun LC-MS/MS technique was used for proteomics identification and Gene Ontology was employed for the bioinformatics analysis. The results of Western blot analysis showed that four proteins bands from jejunal epithelial cells co-cultured with soluble proteins of E. maxima sporozoites were recognized by the positive sera, with molecular weights of 70, 90, 95 and 130 kDa. The co-IP dilutions were analyzed by shotgun LC-MS/MS. A total of 204 proteins were identified in the E. maxima protein database using the MASCOT search engine. Thirty-five proteins including microneme protein 3 and 7 had more than two unique peptide counts and were annotated using Gene Ontology for molecular function, biological process and cellular localization. The results revealed that of the 35 annotated peptides, 22 (62.86%) were associated with binding activity and 15 (42.86%) were involved in catalytic activity. Our findings provide an insight into the interaction between E. maxima and the corresponding host cells and it is important for the understanding of molecular mechanisms underlying E. maxima invasion.

  16. Assessment of probiotics supplementation via feed or water on the growth performance, intestinal morphology and microflora of chickens after experimental infection with Eimeria acervulina, Eimeria maxima and Eimeria tenella.

    Science.gov (United States)

    Giannenas, I; Tsalie, E; Triantafillou, E; Hessenberger, S; Teichmann, K; Mohnl, M; Tontis, D

    2014-01-01

    In this study, the effect of probiotic supplementation via drinking water or feed on the performance of broiler chickens experimentally infected with sporulated oocysts of Eimeria acervulina (5 × 10(4)), Eimeria maxima and Eimeria tenella (2 × 10(4) each one) at 14 days of age was evaluated. Two hundred and forty 1-day-old Ross 308 male chicks were separated into eight equal groups with three replicates. Two of the groups, one infected with mixed Eimeria oocysts and the other not, were given a basal diet and served as controls. The remaining groups were also challenged with mixed Eimeria species and received the basal diet and either water supplemented with probiotic (three groups) or probiotic via feed (two groups); the probiotic used consisted of Enterococcus faecium #589, Bifidobacterium animalis #503 and Lactobacillus salivarius #505 at a ratio of 6:3:1. Probiotic supplementation was applied either via drinking water in different inclusion rates (groups W1, W2 and W3) or via feed using uncoated (group FN) or coated strains (group FC). The last group was given the basal diet supplemented with the anticoccidial lasalocid at 75 mg/kg. Each experimental group was given the corresponding diet or drinking water from day 1 to day 42 of age. Throughout the experimental period of 42 days, body weight and feed intake were recorded weekly and feed conversion ratios were calculated. Seven days after infection, the infected control group presented the lowest weight gain values, while probiotics supplied via feed supported growth to a comparable level with that of the lasalocid group. Probiotic groups presented lesion score values and oocyst numbers that were lower than in control infected birds but higher than in the lasalocid group. In the duodenum, jejunum and ileum, the highest villous height values were presented by probiotic groups. In conclusion, a mixture of probiotic substances gave considerable improvement in both growth performance and intestinal health in

  17. IL-17A regulates Eimeria tenella schizont maturation and migration in avian coccidiosis

    Science.gov (United States)

    Although IL17A is associated with the immunological control of various infectious diseases, its role in host response to Eimeria infections is not well understood. In an effort to better dissect the role of IL17A in host-pathogen interactions in avian coccidiosis, a neutralizing antibody (Ab) to chi...

  18. Anticoccidial effects of coumestans from Eclipta alba for sustainable control of Eimeria tenella parasitosis in poultry production.

    Science.gov (United States)

    Michels, M G; Bertolini, L C T; Esteves, A F; Moreira, P; Franca, S C

    2011-04-19

    The aim of this study was to evaluate the anticoccidial efficacy of a product containing coumestans from Eclipta alba. Experimental conditions were set up as to reproduce the environment conditions for husbandry adopted in commercial broiler farms. Broilers were raised in broiler chicken shed provided with feeders, drinkers, illumination and temperature control systems and floor covering to afford an adequate nourishing environment. Male Cobb broilers (240) were assigned to four experimental groups being each experimental group set apart in rice straw-covered shed isolated with wire mesh. One-day-old broilers were reared in a coccidian-free environment with ad libitum supply of filtered water and freely available standard feed, from the 1st to the 35th day of life. The T1 group received standard feed (negative control); T2 was treated with standard feed supplemented with 66 ppm of salinomycin (positive control); groups T3 and T4 had standard feed supplemented with the ethyl acetate fraction from methanolic extract of E. alba aerial parts, which contains the coumestans WL and DWL (120 and 180 ppm, respectively). The chicken broilers were individually infected with 2 × 104 oocysts of Eimeria tenella when they were 14 days old and were monitored weekly to evaluate zootechnical parameters such as weight gain and food conversion ratio. Counting of coccidial oocyst in chicken feces was assessed from random samples, from the 21st to 28th days of life, which corresponded to 7-14 days after the infection. Five chickens selected at random from each experimental group were subsequently euthanized at 21, 28 or 35 days of life to determine the lesion score in the cecal region and to excise a cecum portion for histopathological evaluation. The group treated with coumestans from E. alba presented an average weight gain and food conversion ratio higher than the negative control group and similar to the mean value of the positive control group. Coumestan-treated groups showed a

  19. IMMUNO-MODULATORY EFFECT OF INACTIVATED EIMERIA TENELLA VACCINE AND LIVE IMPPORTED COCCIDIAL VACCINE ON NEWCASTLE DISEASE VIRUS VACCINA TED BROILER CHICKS

    Directory of Open Access Journals (Sweden)

    Muhammad Akram Muneer, Haji Ahmad Hashmi, Masood Rabbani, Zahid Munir Chaudhry and Ali M. Bahrami

    2001-01-01

    Full Text Available A total of 160 one-day-old broiler chicks were used to evaluate the immunomodulatory effects of an inactivated Eimeria tenella vaccine and a live polyvalent imported antiococcidial vaccine (Coccivac. This study indicated that both of these vaccines did not adversely affect the development of serum antibody against Newcastle disease virus (NDV and the chicks vaccinated with either of the anticoccidial vaccines resisted the virulent NDV challenge. A study of the lymphoid organs such as bursa of fabricuis: thymus and spleen from the experimental chicks indicated that those organs were comparable with those from the chicks not vaccinated with these coccidial vaccines. The overall findings of this study indicate that anticoccidial vaccines do not have any effects on the immune functions of the vaccinates. In fact these vaccines prevented the occurrence of clinical coccidiosis in the vaccinates.

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

    International Nuclear Information System (INIS)

    Hayat, Birjees

    1976-01-01

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

  1. A new adenovirus based vaccine vector expressing an Eimeria tenella derived TLR agonist improves cellular immune responses to an antigenic target.

    Directory of Open Access Journals (Sweden)

    Daniel M Appledorn

    2010-03-01

    Full Text Available Adenoviral based vectors remain promising vaccine platforms for use against numerous pathogens, including HIV. Recent vaccine trials utilizing Adenovirus based vaccines expressing HIV antigens confirmed induction of cellular immune responses, but these responses failed to prevent HIV infections in vaccinees. This illustrates the need to develop vaccine formulations capable of generating more potent T-cell responses to HIV antigens, such as HIV-Gag, since robust immune responses to this antigen correlate with improved outcomes in long-term non-progressor HIV infected individuals.In this study we designed a novel vaccine strategy utilizing an Ad-based vector expressing a potent TLR agonist derived from Eimeria tenella as an adjuvant to improve immune responses from a [E1-]Ad-based HIV-Gag vaccine. Our results confirm that expression of rEA elicits significantly increased TLR mediated innate immune responses as measured by the influx of plasma cytokines and chemokines, and activation of innate immune responding cells. Furthermore, our data show that the quantity and quality of HIV-Gag specific CD8(+ and CD8(- T-cell responses were significantly improved when coupled with rEA expression. These responses also correlated with a significantly increased number of HIV-Gag derived epitopes being recognized by host T cells. Finally, functional assays confirmed that rEA expression significantly improved antigen specific CTL responses, in vivo. Moreover, we show that these improved responses were dependent upon improved TLR pathway interactions.The data presented in this study illustrate the potential utility of Ad-based vectors expressing TLR agonists to improve clinical outcomes dependent upon induction of robust, antigen specific immune responses.

  2. Construction of Lactococcus lactis expressing secreted and anchored Eimeria tenella 3-1E protein and comparison of protective immunity against homologous challenge.

    Science.gov (United States)

    Ma, Chunli; Zhang, Lili; Gao, Mingyang; Ma, Dexing

    2017-07-01

    Two novel plasmids pTX8048-SP-Δ3-1E and pTX8048-SP-NAΔ3-1E-CWA were constructed. The plasmids were respectively electrotransformed into L. lactis NZ9000 to generate strain of L. lactis/pTX8048-SP-Δ3-1E in which 3-1E protein was expressed in secretion, and L. lactis/pTX8048-SP-NAΔ3-1E-CWA on which 3-1E protein was covalently anchored to the surface of bacteria cells. The expression of target proteins were examined by Western blot. The live lactococci expressing secreted 3-1E protein, anchored 3-1E protein, and cytoplasmic 3-1E protein was administered orally to chickens respectively, and the protective immunity and efficacy were compared by animal experiment. The results showed oral immunization to chickens with recombinant lactococci expressing anchored 3-1E protein elicited high 3-1E-specific serum IgG, increased high proportion of CD4 + and CD8α + cells in spleen, alleviated average lesion score in cecum, decreased the oocyst output per chicken compared to lactococci expressing cytoplasmic or secreted 3-1E protein. Taken together, these findings indicated the surface anchored Eimeria protein displayed by L. lacits can induce protective immunity and partial protection against homologous infection. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. From the Macro to the Micro: Gel Mapping to Differentiate between Sporozoites of Two Immunologically Distinct Strains of Eimeria maxima (Strains M6 and Guelph).

    Science.gov (United States)

    El-Ashram, Saeed; Yin, Qing; Liu, Hongbin; Al Nasr, Ibrahim; Liu, Xianyong; Suo, Xun; Barta, John

    2015-01-01

    Two immunologically distinct strains of E. maxima were examined in this study: the M6 strain and the Guelph strain. The differential expression between the sporozoites of the two strains of E. maxima was determined by image analysis of 100 μg of protein from each strain separated by standard one- and conventional two-dimensional polyacrylamide gel electrophoresis. In addition to differences in both molecular weight and the electrophoretic mobility, differences in the intensity of polypeptide bands for example, GS 136.4 and M6 169 were explored. Pooled gels were prepared from each strain. A representative 2D-PAGE gel spanning a non-linear pH range of 3-10 of E. maxima strain M6 consisted of approximately 694 polypeptide spots with about 67 (9.6%) of the polypeptide spots being unique relative to the other strain. E. maxima strain GS had about 696 discernable polypeptide spots with 69 spots (9.9%) that differed from those of the M6 strain. In-depth characterization of the variable polypeptide spots; unique polypeptide spots (absence or presence) and shared polypeptide spots with modifications may lead to novel vaccine target in the form of multi-component, multi-stage, multi-immunovariant strains, multi-species subunit vaccine, and diagnostic probe for E. maxima.

  4. Are Eimeria Genetically Diverse, and Does It Matter?

    Science.gov (United States)

    Clark, Emily L; Tomley, Fiona M; Blake, Damer P

    2017-03-01

    Eimeria pose a risk to all livestock species as a cause of coccidiosis, reducing productivity and compromising animal welfare. Pressure to reduce drug use in the food chain makes the development of cost-effective vaccines against Eimeria essential. For novel vaccines to be successful, understanding genetic and antigenic diversity in field populations is key. Eimeria species that infect chickens are most significant, with Eimeria tenella among the best studied and most economically important. Genome-wide single nucleotide polymorphism (SNP)-based haplotyping has been used to determine population structure, genotype distribution, and potential for cross-fertilization between E. tenella strains. Here, we discuss recent developments in our understanding of diversity for Eimeria in relation to its specialized life cycle, distribution across the globe, and the challenges posed to vaccine development. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Molecular Identification of Eimeria Species in Broiler Chickens in Trinidad, West Indies

    Directory of Open Access Journals (Sweden)

    Arianne Brown Jordan

    2018-01-01

    Full Text Available Coccidiosis is an intestinal disease of chickens of major economic importance to broiler industries worldwide. Species of coccidia found in chickens include Eimeria acervulina, Eimeria brunetti, Eimeria maxima, Eimeria mitis, Eimeria necatrix, Eimeria praecox, and Eimeria tenella. In recent years, polymerase chain reaction (PCR has been developed to provide accurate and rapid identification of the seven known Eimeria species of chickens. The aim of this study was to use species-specific real-time PCR (qPCR to identify which of the seven Eimeria species are present in Trinidad poultry. Seventeen pooled fecal samples were collected from 6 broiler farms (2–5 pens per farm across Trinidad. Feces were also collected from birds showing clinical signs of coccidiosis in two live bird markets (pluck shops. qPCR revealed the presence of five species of Eimeria (E. acervulina, E. maxima, E. mitis, E. necatrix, and E. tenella, but not E. brunetti or E. praecox. Mixed infections were detected on all broiler farms, and DNA of two highly pathogenic Eimeria species (E. tenella and E. necatrix was detected in feces taken from clinically sick birds sampled from the two pluck shops.

  6. Transfection of Eimeria mitis with yellow fluorescent protein as reporter and the endogenous development of the transgenic parasite.

    Directory of Open Access Journals (Sweden)

    Mei Qin

    Full Text Available BACKGROUND: Advancements have been made in the genetic manipulation of apicomplexan parasites. Both the in vitro transient and in vivo stable transfection of Eimeria tenella have been developed successfully. Herein, we report the transient and stable transfection of Eimeria mitis. METHODS AND FINDINGS: Sporozoites of E. mitis transfected with enhanced yellow fluorescent protein (EYFP expression plasmid were inoculated into chickens via the cloacal route. The recovered fluorescent oocysts were sorted by fluorescence activated cell sorting (FACS and then passaged 6 generations successively in chickens. The resulting population was analyzed by genome walking and Western blot. The endogenous development of the transgenic E. mitis was observed and its reproduction potential was tested. The stable transfection of E. mitis was developed. Genome walking confirmed the random integration of plasmid DNA into the genome; while Western blot analysis demonstrated the expression of foreign proteins. Constitutive expression of EYFP was observed in all stages of merogony, gametogony and sporogony. The peak of the transgenic oocyst output was delayed by 24 h and the total oocyst reproduction was reduced by 7-fold when compared to the parental strain. CONCLUSION: Stable transfection of E. mitis was successfully developed. The expression of foreign antigens in the transgenic parasites will facilitate the development of transgenic E. mitis as a vaccine vector.

  7. Prevalence of Eimeria spp. in Broilers by Multiplex PCR in the Southern Region of Brazil on Two Hundred and Fifty Farms.

    Science.gov (United States)

    Moraes, Julio Cesar; França, Marciél; Sartor, Amélia Aparecida; Bellato, Valdomiro; de Moura, Anderson Barbosa; de Lourdes Borba Magalhães, Maria; de Souza, Antonio Pereira; Miletti, Luiz Claudio

    2015-06-01

    Parasitic infections caused by Eimeria species are responsible for most economic losses in poultry production. Prevalence studies can adequately assist the design of prophylaxis strategies for disease control. Therefore, stool samples from 251 flocks of broilers from 28 to 48 days old were collected in 21 municipalities in the state of Santa Catarina, Brazil, to detect and examine the prevalence of Eimeria acervulina, Eimeria maxima, Eimeria tenella, Eimeria mitis, Eimeria praecox, Eimeria necatrix, and Eimeria brunetti. The oocysts were recovered and quantified, and the species were identified by a multiplex PCR technique. Amplicons of seven Eimeria species originating from the PCR-positive samples were cloned. Microscopy studies demonstrated that 96% of the farms were positive for the Eimeria. Seven species were identified, as follows: E. maxima (63.7%) and E. acervulina (63.3%) were the most prevalent species, followed by E. tenella (54.6%), E. mitis (38.6%), E. praecox (25.1%), E. necatrix (24.3%), and E. brunetti (13.1%). The average number of species detected per farm was 2.96, and the most common were E. acervulina, E. maxima, and E. tenella (9.16%). The sequencing of the clones confirmed the specificity and effectiveness of multiplex PCR for the identification of seven species of Eimeria, so this tool can be useful in studying circulating species in poultry farms, thereby assisting prophylactic measures against coccidiosis.

  8. Sporulation dynamics of poultry Eimeria oocysts in Chennai.

    Science.gov (United States)

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

    2015-12-01

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

  9. Plasmodium Sporozoite Biology.

    Science.gov (United States)

    Frischknecht, Friedrich; Matuschewski, Kai

    2017-05-01

    Plasmodium sporozoite transmission is a critical population bottleneck in parasite life-cycle progression and, hence, a target for prophylactic drugs and vaccines. The recent progress of a candidate antisporozoite subunit vaccine formulation to licensure highlights the importance of sporozoite transmission intervention in the malaria control portfolio. Sporozoites colonize mosquito salivary glands, migrate through the skin, penetrate blood vessels, breach the liver sinusoid, and invade hepatocytes. Understanding the molecular and cellular mechanisms that mediate the remarkable sporozoite journey in the invertebrate vector and the vertebrate host can inform evidence-based next-generation drug development programs and immune intervention strategies. Copyright © 2017 Cold Spring Harbor Laboratory Press; all rights reserved.

  10. Prevalence and distribution of Eimeria species in broiler chicken farms of different capacities

    Science.gov (United States)

    Györke, Adriana; Pop, Loredana; Cozma, Vasile

    2013-01-01

    We conducted a survey in broiler farms from Romania to establish prevalence and distribution of Eimeria species using single PCR assay. We found Eimeria spp. in 21 (91%) out of 23 flocks, and in 11 (92%) out of 12 farms. Four species of Eimeria were identified: E. acervulina (21/23; 91%), E. tenella (14/23; 61%), E. maxima (5/23; 22%) and E. praecox (3/23; 13%). Infection with a single species (E. acervulina) was detected in 6 (26%) infected flocks originated from large farms. Mixed infections were found in 15 (65%) flocks and the most prevalent combination was E. acervulina + E. tenella (8/23; 35%). Four flocks (17%) harboured mixed infection with E. acervulina + E. tenella + E. maxima. E. acervulina was significantly more prevalent in flocks that received ionophores as anticoccidial feed additives. PMID:24309007

  11. The Eimeria Transcript DB: an integrated resource for annotated transcripts of protozoan parasites of the genus Eimeria

    Science.gov (United States)

    Rangel, Luiz Thibério; Novaes, Jeniffer; Durham, Alan M.; Madeira, Alda Maria B. N.; Gruber, Arthur

    2013-01-01

    Parasites of the genus Eimeria infect a wide range of vertebrate hosts, including chickens. We have recently reported a comparative analysis of the transcriptomes of Eimeria acervulina, Eimeria maxima and Eimeria tenella, integrating ORESTES data produced by our group and publicly available Expressed Sequence Tags (ESTs). All cDNA reads have been assembled, and the reconstructed transcripts have been submitted to a comprehensive functional annotation pipeline. Additional studies included orthology assignment across apicomplexan parasites and clustering analyses of gene expression profiles among different developmental stages of the parasites. To make all this body of information publicly available, we constructed the Eimeria Transcript Database (EimeriaTDB), a web repository that provides access to sequence data, annotation and comparative analyses. Here, we describe the web interface, available sequence data sets and query tools implemented on the site. The main goal of this work is to offer a public repository of sequence and functional annotation data of reconstructed transcripts of parasites of the genus Eimeria. We believe that EimeriaTDB will represent a valuable and complementary resource for the Eimeria scientific community and for those researchers interested in comparative genomics of apicomplexan parasites. Database URL: http://www.coccidia.icb.usp.br/eimeriatdb/ PMID:23411718

  12. Molecular characterization of eimeria species naturally infecting egyptian baldi chickens.

    Directory of Open Access Journals (Sweden)

    Sahar M Gadelhaq

    2015-03-01

    Full Text Available Coccidiosis is a serious protozoal disease of poultry. The identification of Eimeria species has important implications for diagnosis and control as well as for epidemiology. The molecular characterization of Eimeria species infecting Egyptian baladi chickens was investigated.Eimeria species oocysts were harvested from intestines of naturally infected Egyptian baldi chickens. The morphometry characterization of oocysts along with COCCIMORPH software was done. The DNA was extracted initially by freezing and thawing then the prepared samples was subjected to commercial DNA kits. The DNA products were analyzed through conventional polymerase chain reaction by using amplified region (SCAR marker.The PCR results confirmed the presence of 7 Eimeria species in the examined fecal samples of Egyptian baldi breed with their specific ampilicon sizes being E. acervulina (811bp, E. brunette (626bp, E. tenella (539bp, E. maxima (272bp, E. necatrix (200bp, E. mitis (327bp and E. praecopx (354bp. A sequencing of the two most predominant species of Eimeria was done, on E. tenella and E. máxima. Analysis of the obtained sequences revealed high identities 99% between Egyptian isolates and the reference one. Similarly, E. maxima isolated from Egyptian baldi chickens showed 98% nucleotide identities with the reference strain. Only single nucleotide substitution was observed among the Egyptian E. tenella isolates (A181G when compared to the reference one. The Egyptian isolates acquired 4 unique mutations (A68T, C164T, G190A and C227G in compared with the reference sequence.This is the first time to identify the 7 species of Eimeria from Egyptian baladi chickens.

  13. Molecular characterization of eimeria species naturally infecting egyptian baldi chickens.

    Science.gov (United States)

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

    2015-01-01

    Coccidiosis is a serious protozoal disease of poultry. The identification of Eimeria species has important implications for diagnosis and control as well as for epidemiology. The molecular characterization of Eimeria species infecting Egyptian baladi chickens was investigated. Eimeria species oocysts were harvested from intestines of naturally infected Egyptian baldi chickens. The morphometry characterization of oocysts along with COCCIMORPH software was done. The DNA was extracted initially by freezing and thawing then the prepared samples was subjected to commercial DNA kits. The DNA products were analyzed through conventional polymerase chain reaction by using amplified region (SCAR) marker. The PCR results confirmed the presence of 7 Eimeria species in the examined fecal samples of Egyptian baldi breed with their specific ampilicon sizes being E. acervulina (811bp), E. brunette (626bp), E. tenella (539bp), E. maxima (272bp), E. necatrix (200bp), E. mitis (327bp) and E. praecopx (354bp). A sequencing of the two most predominant species of Eimeria was done, on E. tenella and E. máxima. Analysis of the obtained sequences revealed high identities 99% between Egyptian isolates and the reference one. Similarly, E. maxima isolated from Egyptian baldi chickens showed 98% nucleotide identities with the reference strain. Only single nucleotide substitution was observed among the Egyptian E. tenella isolates (A181G) when compared to the reference one. The Egyptian isolates acquired 4 unique mutations (A68T, C164T, G190A and C227G) in compared with the reference sequence. This is the first time to identify the 7 species of Eimeria from Egyptian baladi chickens.

  14. Molecular Characterization of Eimeria Species Naturally Infecting Egyptian Baldi Chickens

    Science.gov (United States)

    GADELHAQ, Sahar M; ARAFA, Waleed M; ABOELHADID, Shawky M

    2015-01-01

    Background: Coccidiosis is a serious protozoal disease of poultry. The identification of Eimeria species has important implications for diagnosis and control as well as for epidemiology. The molecular characterization of Eimeria species infecting Egyptian baladi chickens was investigated. Methods: Eimeria species oocysts were harvested from intestines of naturally infected Egyptian baldi chickens. The morphometry characterization of oocysts along with COCCIMORPH software was done. The DNA was extracted initially by freezing and thawing then the prepared samples was subjected to commercial DNA kits. The DNA products were analyzed through conventional polymerase chain reaction by using amplified region (SCAR) marker. Results: The PCR results confirmed the presence of 7 Eimeria species in the examined fecal samples of Egyptian baldi breed with their specific ampilicon sizes being E. acervulina (811bp), E. brunette (626bp), E. tenella (539bp), E. maxima (272bp), E. necatrix (200bp), E. mitis (327bp) and E. praecopx (354bp). A sequencing of the two most predominant species of Eimeria was done, on E. tenella and E. máxima. Analysis of the obtained sequences revealed high identities 99% between Egyptian isolates and the reference one. Similarly, E. maxima isolated from Egyptian baldi chickens showed 98% nucleotide identities with the reference strain. Only single nucleotide substitution was observed among the Egyptian E. tenella isolates (A181G) when compared to the reference one. The Egyptian isolates acquired 4 unique mutations (A68T, C164T, G190A and C227G) in compared with the reference sequence. Conclusion: This is the first time to identify the 7 species of Eimeria from Egyptian baladi chickens. PMID:25904950

  15. Complete mitochondrial genome sequences from five Eimeria species (Apicomplexa; Coccidia; Eimeriidae) infecting domestic turkeys.

    Science.gov (United States)

    Ogedengbe, Mosun E; El-Sherry, Shiem; Whale, Julia; Barta, John R

    2014-07-17

    via host switching from another avian host. Phylogenetic analyses suggest E. necatrix and E. tenella are related distantly to other Eimeria of chickens. Mitochondrial genomes of Eimeria species sequenced to date are highly conserved with regard to gene content and structure. Nonetheless, complete mitochondrial genome sequences and, particularly the three CDS, possess sufficient sequence variability for differentiating Eimeria species of poultry. The mitochondrial genome sequences are highly suited for molecular diagnostics and phylogenetics of coccidia and, potentially, genetic markers for molecular epidemiology.

  16. Expression of avian beta-defensins in the intestine of Eimeria-challenged chickens

    Science.gov (United States)

    Avian coccidiosis is caused by the intracellular protozoa Eimeria. The site of invasion and lesions in the intestine is species-specific; for example, E. acervulina mainly affects the duodenum, E. maxima the jejunum, and E. tenella the ceca. Lesions in the intestinal mucosa reduce feed efficiency a...

  17. Expression of digestive enzymes and nutrient transporters in Eimeria-challenged broilers

    Science.gov (United States)

    Avian coccidiosis is a disease caused by the intestinal protozoa Eimeria. The site of invasion and lesions in the intestine is species-specific, for example E. acervulina affects the duodenum, E. maxima the jejunum, and E. tenella the ceca. Lesions in the intestinal mucosa cause reduced feed effic...

  18. 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.

  19. Cryptic Eimeria genotypes are common across the southern but not northern hemisphere.

    Science.gov (United States)

    Clark, Emily L; Macdonald, Sarah E; Thenmozhi, V; Kundu, Krishnendu; Garg, Rajat; Kumar, Saroj; Ayoade, Simeon; Fornace, Kimberly M; Jatau, Isa Danladi; Moftah, Abdalgader; Nolan, Matthew J; Sudhakar, N R; Adebambo, A O; Lawal, I A; Álvarez Zapata, Ramón; Awuni, Joseph A; Chapman, H David; Karimuribo, Esron; Mugasa, Claire M; Namangala, Boniface; Rushton, Jonathan; Suo, Xun; Thangaraj, Kumarasamy; Srinivasa Rao, Arni S R; Tewari, Anup K; Banerjee, Partha S; Dhinakar Raj, G; Raman, M; Tomley, Fiona M; Blake, Damer P

    2016-08-01

    The phylum Apicomplexa includes parasites of medical, zoonotic and veterinary significance. Understanding the global distribution and genetic diversity of these protozoa is of fundamental importance for efficient, robust and long-lasting methods of control. Eimeria spp. cause intestinal coccidiosis in all major livestock animals and are the most important parasites of domestic chickens in terms of both economic impact and animal welfare. Despite having significant negative impacts on the efficiency of food production, many fundamental questions relating to the global distribution and genetic variation of Eimeria spp. remain largely unanswered. Here, we provide the broadest map yet of Eimeria occurrence for domestic chickens, confirming that all the known species (Eimeria acervulina, Eimeria brunetti, Eimeria maxima, Eimeria mitis, Eimeria necatrix, Eimeria praecox, Eimeria tenella) are present in all six continents where chickens are found (including 21 countries). Analysis of 248 internal transcribed spacer sequences derived from 17 countries provided evidence of possible allopatric diversity for species such as E. tenella (FST values ⩽0.34) but not E. acervulina and E. mitis, and highlighted a trend towards widespread genetic variance. We found that three genetic variants described previously only in Australia and southern Africa (operational taxonomic units x, y and z) have a wide distribution across the southern, but not the northern hemisphere. While the drivers for such a polarised distribution of these operational taxonomic unit genotypes remains unclear, the occurrence of genetically variant Eimeria may pose a risk to food security and animal welfare in Europe and North America should these parasites spread to the northern hemisphere. Copyright © 2016 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  20. Translational Repression in Malaria Sporozoites

    Science.gov (United States)

    Turque, Oliver; Tsao, Tiffany; Li, Thomas; Zhang, Min

    2016-01-01

    Malaria is a mosquito-borne infectious disease of humans and other animals. It is caused by the parasitic protozoan, Plasmodium. Sporozoites, the infectious form of malaria parasites, are quiescent when they remain in the salivary glands of the Anopheles mosquito until transmission into a mammalian host. Metamorphosis of the dormant sporozoite to its active form in the liver stage requires transcriptional and translational regulations. Here, we summarize recent advances in the translational repression of gene expression in the malaria sporozoite. In sporozoites, many mRNAs that are required for liver stage development are translationally repressed. Phosphorylation of eukaryotic Initiation Factor 2α (eIF2α) leads to a global translational repression in sporozoites. The eIF2α kinase, known as Upregulated in Infectious Sporozoite 1 (UIS1), is dominant in the sporozoite. The eIF2α phosphatase, UIS2, is translationally repressed by the Pumilio protein Puf2. This translational repression is alleviated when sporozoites are delivered into the mammalian host. PMID:28357358

  1. Translational repression in malaria sporozoites

    Directory of Open Access Journals (Sweden)

    Oliver Turque

    2016-04-01

    Full Text Available Malaria is a mosquito-borne infectious disease of humans and other animals. It is caused by the parasitic protozoan, Plasmodium. Sporozoites, the infectious form of malaria parasites, are quiescent when they remain in the salivary glands of the Anopheles mosquito until transmission into a mammalian host. Metamorphosis of the dormant sporozoite to its active form in the liver stage requires transcriptional and translational regulations. Here, we summarize recent advances in the translational repression of gene expression in the malaria sporozoite. In sporozoites, many mRNAs that are required for liver stage development are translationally repressed. Phosphorylation of eukaryotic Initiation Factor 2α (eIF2α leads to a global translational repression in sporozoites. The eIF2α kinase, known as Upregulated in Infectious Sporozoite 1 (UIS1, is dominant in the sporozoite. The eIF2α phosphatase, UIS2, is translationally repressed by the Pumilio protein Puf2. This translational repression is alleviated when sporozoites are delivered into the mammalian host.

  2. Oral treatment of Eimeria tenella -infected broilers using aqueous ...

    African Journals Online (AJOL)

    In Nigeria, wild Ganoderma species of mushroom grows in abundance during the rainy season. Studies were conducted to evaluate the haematological parameters and the histopathology lesions in organs of broilers treated with aqueous extract of wild Ganoderma sp. Blood and organs were collected for haematology and ...

  3. Meta-analysis of the performance variation in broilers experimentally challenged by Eimeria spp.

    Science.gov (United States)

    Kipper, Marcos; Andretta, Ines; Lehnen, Cheila Roberta; Lovatto, Paulo Alberto; Monteiro, Silvia Gonzalez

    2013-09-01

    A meta-analysis was carried out to (1) study the relation of the variation in feed intake and weight gain in broilers infected with Eimeria acervulina, Eimeria maxima, Eimeria tenella, or a Pool of Eimeria species, and (2) to identify and to quantify the effects involved in the infection. A database of articles addressing the experimental infection with Coccidia in broilers was developed. These publications must present results of animal performance (weight gain, feed intake, and feed conversion ratio). The database was composed by 69 publications, totalling around 44 thousand animals. Meta-analysis followed three sequential analyses: graphical, correlation, and variance-covariance. The feed intake of the groups challenged by E. acervulina and E. tenella did not differ (P>0.05) to the control group. However, the feed intake in groups challenged by E. maxima and Pool showed an increase of 8% and 5% (PEimeria species, animal age, sex, and genetic line. In general the age effect is superior to the challenge effect, showing that age at the challenge is important to determine the impact of Eimeria infection. Copyright © 2013 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2017-06-01

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

  5. Detection of four important Eimeria species by multiplex PCR in a single assay.

    Science.gov (United States)

    You, Myung-Jo

    2014-06-01

    The oocysts of some of the recognized species of chicken coccidiosis are difficult to distinguish morphologically. Diagnostic laboratories are increasingly utilizing DNA-based technologies for the specific identification of Eimeria species. This study reports a multiplex polymerase chain reaction (PCR) assay based on internal transcribed spacer-1 (ITS-1) for the simultaneous diagnosis of the Eimeria tenella, Eimeria acervulina, Eimeria maxima, and Eimeria necatrix species, which infect domestic fowl. Primer pairs specific to each species were designed in order to generate a ladder of amplification products ranging from 20 to 25 bp, and a common optimum annealing temperature for these species was determined to be 52.5 °C. Sensitivity tests were performed for each species, showing a detection threshold of 1-5 pg. All the species were amplified homogeneously, and a homogenous band ladder was observed, indicating that the assay permitted the simultaneous detection of all the species in a single-tube reaction. In the phylogenic study, there was a clear species clustering, which was irrespective of geographical location, for all the ITS-1 sequences used. This multiplex PCR assay represents a rapid and potential cost-effective diagnostic method for the detection of some key Eimeria species that infect domestic fowl. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  6. Eimeria pipistrellus n. sp. from Pipistrellus kuhlii (Chiroptera: Vespertilionidae) in Saudi Arabia

    Science.gov (United States)

    Al-Dakhil, Mohamed; Al-Shawa, Yaser

    1999-01-01

    Fecal samples from 12 Pipistrellus kuhlii captured at Shagrah, Saudi Arabia, were examined for coccidia and three (25%) found to harbor a undescribed eimerian, herein described as Eimeria pipistrellus n. sp. Sporulated oocysts were subspherical, 24.8×23.2 (22-27×20-25) µm, with a bilayered and smooth wall. The micropyle was absent, but a large oocyst residuum and a single polar granule were present. Sporocysts were ovoid, 11.6×8.3 (10.5-13×7.5-9) µm, with a prominent Stieda body, but without a substiedal body; sporozoites lay head to tail in sporocysts and contained one large posterior refractile body. Eimeria pipistrellus n. sp. is the 3rd species of the genus Eimeria found from bats of the genus Pipistrellus. PMID:10188376

  7. Description of Eimeria motelo sp. n. (Apicomplexa: Eimeriidae from the yellow footed tortoise, Geochelone denticulata (Chelonia: Testudinidae, and replacement of Eimeria carinii Lainson, Costa & Shaw, 1990 by Eimeria lainsoni nom. nov.

    Directory of Open Access Journals (Sweden)

    Lada Hurková

    2000-12-01

    Full Text Available Eimeria motelo sp. n. is described from faeces of the yellow-footed tortoise, Geochelone denticulata (L.. Oocysts are irregularly ellipsoidal or cylindrical, with slightly expressed lobed protrusions and irregularities at the poles, possibly caused by wrinkling of the oocyst wall, 17 (15-19 × 9.4 (8.5-11 µm, shape index (length/width being 1.81 (1.45-2. The oocyst wall is smooth, single-layered, 0.5 µm thick with no micropyle. There are no polar bodies. Sporocysts are ellipsoidal, 8.9 (7.5-10 × 4.4 (4-5 µm, shape index 2.03 (1.7-2.5. A sporocyst residuum is present, composed of many granules of irregular size. The sporozoites are elongate, lying lengthwise in the sporocysts. Comparison with other species of the genus Eimeria parasitising members of family Testudinidae indicates that the presently described coccidium represents a new species. The name of Eimeria carinii Lainson, Costa & Shaw, 1990 is found to be preoccupied by a homonym, Eimeria carinii Pinto 1928 given to a coccidium from Rattus norvegicus. Therefore, it is replaced by Eimeria lainsoni nom. nov.

  8. Further characterisation of two Eimeria species (Eimeria quokka and Eimeria setonicis) in quokkas (Setonix brachyurus).

    Science.gov (United States)

    Austen, J M; Friend, J A; Yang, R; Ryan, U M

    2014-03-01

    The identification and characterisation of novel Eimeria species has largely been based on sporulated oocyst and sporocyst morphology, the host species and the geographical range. Variation in the size and shape of Eimeria oocysts across their host range however, make the identification and characterisation of novel species using traditional methodologies alone problematic. The use of molecular markers and phylogenetic analysis has greatly advanced our ability to characterise Eimeria species and has recently been applied to understand evolutionary relationships among Eimeria species from Australian marsupials. In the present study, Eimeria species isolated from quokkas (Setonix brachyurus) captured from Two Peoples Bay, Bald Island and Rottnest Island, Western Australia, were morphologically identified as Eimeria quokka and Eimeria setonicis. Both Eimeria species were identified as being polymorphic in nature with regards to sporulated oocyst and sporocyst morphometrics. Phylogenetic analysis using 18S rRNA and COI (cytochrome c oxidase subunit 1) genes, grouped E. quokka and E. setonicis within the Eimeria marsupial clade together with Eimeria trichosuri from brushtail possums, Eimeria macropodis from tammar wallabies (Macropus eugenii) and several unidentified macropod Eimeria species from western grey kangaroos (Macropus fuliginosus). This study is the first to characterise E. quokka and E. setonicis by molecular analysis, enabling more extensive resolution of evolutionary relationships among marsupial-derived Eimeria species. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. Epidemiology of Eimeria species in selected broiler farms of Khoy suburb, West Azarbaijan Province, Iran

    Directory of Open Access Journals (Sweden)

    Fakhri, M.

    2015-12-01

    Full Text Available Intestinal coccidiosis, caused by Eimeria species, is an economically-important disease of poultry production industry worldwide. This study was designed to investigate the prevalence of different Eimeria species in the farmed broilers of Khoy city, West Azarbaijan, North West Iran. A total of 26 broiler farms of different production capacities were arbitrarily selected and examined in 2013. In each of the farms, Litters of two broilers farms were randomly sampled twice a week and examined. The intensity of infection with each of the Eimeria species was assessed on the basis of number of oocysts per gram of litter using Clayton-Lane and McMaster methods. Eimeria species diversity was determined by using oocyst sporulation technique in 2% potassium dichromate solution. Results indicated that 23.08% (6/26 of the broiler farms were infected with Eimeria oocysts. The maximum litter infection rate (7.5×103 was observed in fifth week of the rearing period. The litter infection rate was significantly correlated with kinds of water dispenser, feeder, ventilation, and density. The litters were infected with five Eimeria species; E. maxima (32.67% in 6 farms (23.07%, E. mitis (24% in 6 farms (23.07%, E. acervulina (18% in 5 farms (19.23%, E. tenella (14.67% in 4 farms (15.38%, and E. necatrix (10.67% in 3 farms (11.58%. Results of this study uncovered high rates of litter infection with various Eimeria species in the studied farms, suggesting the establishment of firm health management strategies in the region.

  10. 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.

  11. Eimeria species occurrence varies between geographic regions and poultry production systems and may influence parasite genetic diversity.

    Science.gov (United States)

    Chengat Prakashbabu, B; Thenmozhi, V; Limon, G; Kundu, K; Kumar, S; Garg, R; Clark, E L; Srinivasa Rao, A S R; Raj, D G; Raman, M; Banerjee, P S; Tomley, F M; Guitian, J; Blake, D P

    2017-01-15

    Coccidiosis is one of the biggest challenges faced by the global poultry industry. Recent studies have highlighted the ubiquitous distribution of all Eimeria species which can cause this disease in chickens, but intriguingly revealed a regional divide in genetic diversity and population structure for at least one species, Eimeria tenella. The drivers associated with such distinct geographic variation are unclear, but may impact on the occurrence and extent of resistance to anticoccidial drugs and future subunit vaccines. India is one of the largest poultry producers in the world and includes a transition between E. tenella populations defined by high and low genetic diversity. The aim of this study was to identify risk factors associated with the prevalence of Eimeria species defined by high and low pathogenicity in northern and southern states of India, and seek to understand factors which vary between the regions as possible drivers for differential genetic variation. Faecal samples and data relating to farm characteristics and management were collected from 107 farms from northern India and 133 farms from southern India. Faecal samples were analysed using microscopy and PCR to identify Eimeria occurrence. Multiple correspondence analysis was applied to transform correlated putative risk factors into a smaller number of synthetic uncorrelated factors. Hierarchical cluster analysis was used to identify poultry farm typologies, revealing three distinct clusters in the studied regions. The association between clusters and presence of Eimeria species was assessed by logistic regression. The study found that large-scale broiler farms in the north were at greatest risk of harbouring any Eimeria species and a larger proportion of such farms were positive for E. necatrix, the most pathogenic species. Comparison revealed a more even distribution for E. tenella across production systems in south India, but with a lower overall occurrence. Such a polarised region- and

  12. Effects of in ovo supplementation of probiotics on performance and immunocompetence of broiler chicks to an Eimeria challenge.

    Science.gov (United States)

    Pender, C M; Kim, S; Potter, T D; Ritzi, M M; Young, M; Dalloul, R A

    2016-11-30

    Coccidiosis is regarded as the parasitic disease with the greatest economic impact on the poultry industry due to reduced performance and increased mortality. A study was conducted to investigate the effects of in ovo administration of probiotics on hatchability, performance, immune organ weights, and lesion scores in broiler chicks during a mixed Eimeria infection. At embryonic day 18, 210 eggs were injected with either sterile water or 1×10 6 cfu probiotic bacteria. On day 3 post-hatch, half of the chicks from each treatment group were challenged with a mixed inoculum of Eimeria acervulina, Eimeria maxima and Eimeria tenella. Measurements and tissue samples were taken on day of hatch (DOH) and days 3, 9 and 15. On day 9, 24 birds per treatment were scored for intestinal Eimeria lesions. No differences were seen among groups for hatchability as well as for body weight (BW), BW gain (BWG), or immune organ weights prior to the Eimeria challenge. On day 9, the non-challenged birds with probiotic supplementation had higher BW and BWG than the non-supplemented controls while no differences were seen among the challenged groups. On day 15, probiotic supplemented birds had improved BW compared to the non-supplemented birds as well as increased BWG from day 9 to 15. Bursa weight was not affected by treatment at any time point while spleen weight was greater in supplemented birds on day 15. Birds receiving the probiotic had significantly lower mortality than non-treated birds. Additionally, gross lesion severity was reduced due to probiotic supplementation in all intestinal segments evaluated. These results suggest that in ovo supplementation of probiotics may improve early performance and provide protection against a mixed Eimeria infection.

  13. Host response to Eimeria infections

    NARCIS (Netherlands)

    Swinkels, W.J.C.

    2008-01-01

    The protozoan parasite Eimeria is responsible for the disease coccidiosis and has a worldwide distribution. Intestinal Eimeria infections are the dominating class of diseases in poultry causing great economical damage and considerably affecting animal welfare. In the Netherlands in chickens raised

  14. Eimeria genomics: Where are we now and where are we going?

    Science.gov (United States)

    Blake, Damer P

    2015-08-15

    The evolution of sequencing technologies, from Sanger to next generation (NGS) and now the emerging third generation, has prompted a radical frameshift moving genomics from the specialist to the mainstream. For parasitology, genomics has moved fastest for the protozoa with sequence assemblies becoming available for multiple genera including Babesia, Cryptosporidium, Eimeria, Giardia, Leishmania, Neospora, Plasmodium, Theileria, Toxoplasma and Trypanosoma. Progress has commonly been slower for parasites of animals which lack zoonotic potential, but the deficit is now being redressed with impact likely in the areas of drug and vaccine development, molecular diagnostics and population biology. Genomics studies with the apicomplexan Eimeria species clearly illustrate the approaches and opportunities available. Specifically, more than ten years after initiation of a genome sequencing project a sequence assembly was published for Eimeria tenella in 2014, complemented by assemblies for all other Eimeria species which infect the chicken and Eimeria falciformis, a parasite of the mouse. Public access to these and other coccidian genome assemblies through resources such as GeneDB and ToxoDB now promotes comparative analysis, encouraging better use of shared resources and enhancing opportunities for development of novel diagnostic and control strategies. In the short term genomics resources support development of targeted and genome-wide genetic markers such as single nucleotide polymorphisms (SNPs), with whole genome re-sequencing becoming viable in the near future. Experimental power will develop rapidly as additional species, strains and isolates are sampled with particular emphasis on population structure and allelic diversity. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Expression of host defense peptides in the intestine of Eimeria-challenged chickens.

    Science.gov (United States)

    Su, S; Dwyer, D M; Miska, K B; Fetterer, R H; Jenkins, M C; Wong, E A

    2017-07-01

    Avian coccidiosis is caused by the intracellular protozoan Eimeria, which produces intestinal lesions leading to weight gain depression. Current control methods include vaccination and anticoccidial drugs. An alternative approach involves modulating the immune system. The objective of this study was to profile the expression of host defense peptides such as avian beta-defensins (AvBDs) and liver expressed antimicrobial peptide 2 (LEAP2), which are part of the innate immune system. The mRNA expression of AvBD family members 1, 6, 8, 10, 11, 12, and 13 and LEAP2 was examined in chickens challenged with either E. acervulina, E. maxima, or E. tenella. The duodenum, jejunum, ileum, and ceca were collected 7 d post challenge. In study 1, E. acervulina challenge resulted in down-regulation of AvBD1, AvBD6, AvBD10, AvBD11, AvBD12, and AvBD13 in the duodenum. E. maxima challenge caused down-regulation of AvBD6, AvBD10, and AvBD11 in the duodenum, down-regulation of AvBD10 in the jejunum, but up-regulation of AvBD8 and AvBD13 in the ceca. E. tenella challenge showed no change in AvBD expression in any tissue. In study 2, which involved challenge with only E. maxima, there was down-regulation of AvBD1 in the ileum, AvBD11 in the jejunum and ileum, and LEAP2 in all 3 segments of the small intestine. The expression of LEAP2 was further examined by in situ hybridization in the jejunum of chickens from study 2. LEAP2 mRNA was expressed similarly in the enterocytes lining the villi, but not in the crypts of control and Eimeria challenged chickens. The lengths of the villi in the Eimeria challenged chickens were less than those in the control chickens, which may in part account for the observed down-regulation of LEAP2 mRNA quantified by PCR. Overall, the AvBD response to Eimeria challenge was not consistent; whereas LEAP2 was consistently down-regulated, which suggests that LEAP2 plays an important role in modulating an Eimeria infection. Published by Oxford University Press on

  16. Divergent nuclear 18S rDNA paralogs in a turkey coccidium, Eimeria meleagrimitis, complicate molecular systematics and identification.

    Science.gov (United States)

    El-Sherry, Shiem; Ogedengbe, Mosun E; Hafeez, Mian A; Barta, John R

    2013-07-01

    Multiple 18S rDNA sequences were obtained from two single-oocyst-derived lines of each of Eimeria meleagrimitis and Eimeria adenoeides. After analysing the 15 new 18S rDNA sequences from two lines of E. meleagrimitis and 17 new sequences from two lines of E. adenoeides, there were clear indications that divergent, paralogous 18S rDNA copies existed within the nuclear genome of E. meleagrimitis. In contrast, mitochondrial cytochrome c oxidase subunit I (COI) partial sequences from all lines of a particular Eimeria sp. were identical and, in phylogenetic analyses, COI sequences clustered unambiguously in monophyletic and highly-supported clades specific to individual Eimeria sp. Phylogenetic analysis of the new 18S rDNA sequences from E. meleagrimitis showed that they formed two distinct clades: Type A with four new sequences; and Type B with nine new sequences; both Types A and B sequences were obtained from each of the single-oocyst-derived lines of E. meleagrimitis. Together these rDNA types formed a well-supported E. meleagrimitis clade. Types A and B 18S rDNA sequences from E. meleagrimitis had a mean sequence identity of only 97.4% whereas mean sequence identity within types was 99.1-99.3%. The observed intraspecific sequence divergence among E. meleagrimitis 18S rDNA sequence types was even higher (approximately 2.6%) than the interspecific sequence divergence present between some well-recognized species such as Eimeria tenella and Eimeria necatrix (1.1%). Our observations suggest that, unlike COI sequences, 18S rDNA sequences are not reliable molecular markers to be used alone for species identification with coccidia, although 18S rDNA sequences have clear utility for phylogenetic reconstruction of apicomplexan parasites at the genus and higher taxonomic ranks. Copyright © 2013. Published by Elsevier Ltd.

  17. A NEW SPECIES OF EIMERIA (APICOMPLEXA: EIMERIIDAE) FROM THE NORTHERN MYOTIS, MYOTIS SEPTENTRIONALIS (CHIROPTERA: VESPERTILIONIDAE), IN OKLAHOMA

    Science.gov (United States)

    McAllister, Chris T.; Seville, R. Scott; Roehrs, Zachary P.

    2012-01-01

    During September 2004, 4 adult northern myotis, Myotis septentrionalis, were collected from LeFlore County, Oklahoma (n = 2), and Logan (n = 1) and Yell (n = 1) counties, Arkansas, and their feces examined for coccidian parasites. Three of 4 bats (75%) were passing oocysts of Eimeria spp. Oocysts of Eimeria tumlisoni n. sp. were ovoidal, 17.6 × 16.8 (16–19 × 14–18) μm with a shape index of 1.0 (1.0–1.1). A micropyle and oocyst residuum were absent, although 1–2 bilobed polar granules were often present. Sporocysts were ovoidal, 10.5 × 5.9 (9–12 × 5–7) μm with a shape index of 1.8 (1.6–2.0). A Stieda body was present, but sub–Stieda and para–Stieda bodies were absent. A sporocyst residuum was present consisting of compact to dispersed granules between the sporozoites. The sporozoites were elongate, with subspherical anterior refractile body and spherical posterior refractile body; a nucleus was not discernable. This is the second coccidian reported from this host and the first instance of a bat coccidian reported from Oklahoma. We also document a new geographic record for Eimeria catronensis in Oklahoma, and provide an emended description. PMID:22509940

  18. Two new Eimeria species (Apicomplexa: Eimeriidae) from the yellow-crowned Amazon Amazona ochrocephala (Aves: Psittacidae) in Brazil.

    Science.gov (United States)

    Hofstatter, P G; Kawazoe, U

    2011-06-01

    In this study, we describe 2 new species of Eimeria associated with the yellow-crowned Amazon Amazona ochrocephala. Eimeria amazonae n. sp. has bilayered, ellipsoidal, and smooth oocysts that measure 48.9 × 36.2 µm; the length/width ratio is 1.35. The micropyle and oocyst residuum are both absent, but the polar granule is present. Ovoidal sporocysts are 22.2 × 11.9 µm. Stieda and sub-Stieda bodies and sporocyst residuum are present. The 2 elongate sporozoites are curved and measure 18.1 × 3.4 µm; both have 2 refractile bodies. Eimeria ochrocephalae n. sp. has bilayered, ellipsoidal, and smooth oocysts that measure 43.8 × 27.7 µm; the length/width ratio is 1.58. The micropyle and oocyst residuum are absent, but the polar granule is present; ovoidal sporocysts are 20.6 × 10.1 µm. Stieda and sub-Stieda bodies and sporocyst residuum are present; 2 elongate and curved sporozoites are 15.8 × 3.4 µm, each of which has 2 refractile bodies.

  19. Re-evaluation of the life cycle of Eimeria maxima Tyzzer, 1929 in chickens (Gallus domesticus).

    Science.gov (United States)

    Dubey, J P; Jenkins, M C

    2017-12-14

    A time-course study was conducted to resolve discrepancies in the literature and better define aspects of the Eimeria maxima life cycle such, as sites of development and both morphology and number of asexual stages. Broiler chickens were inoculated orally with five million E. maxima oocysts (APU1), and were necropsied at regular intervals from 12 to 120 h p.i. Small intestine tissue sections and smears were examined for developmental stages. The jejunum contained the highest numbers of developmental stages. At 12 h p.i., sporozoites were observed inside a parasitophorous vacuole (PV) in the epithelial villi and the lamina propria. By 24 h, sporozoites enclosed by a PV were observed in enterocytes of the glands of Lieberkühn. At 48 h p.i., sporozoites, elongated immature and mature schizonts, were all seen in the glands with merozoites budding off from a residual body. By 60 h, second-generation, sausage-shaped schizonts containing up to 12 merozoites were observed around a residual body in the villar tip of invaded enterocytes. At 72 and 96 h, profuse schizogony associated with third- and fourth-generation schizonts was observed throughout the villus. At 120 h, another generation (fifth) of schizonts were seen in villar tips as well as in subepithelium where gamonts and oocysts were also present; a few gamonts were in epithelium. Our finding of maximum parasitization of E. maxima in jejunum is important because this region is critical for nutrient absorption and weight gain.

  20. Eimeria jayakaris sp. n. (Apicomplexa:Eimeriidae) from the snake, Eryx jayakari (Serpentes: Boidae) in Saudi Arabia

    International Nuclear Information System (INIS)

    Alyousif, Mohamed S; AlShawa, Yaser R

    2007-01-01

    A new coccidian parasite of the genus Eimeria is described from the gall bladder of the boid snake, Eryx jayakari collected from Althumamah, central region, Saudi Arabia. Oocysts of Eimeria jayakaris sp. n. are ellipsoid, measuring 31x19.5 (28.7-33.5 x 18.5-20.8) micron meter, with a smooth greenish-yellow bilayered oocyst wall of 1.1 (0.9-1.2) micron meter. Micropyle, oocyst residuum and polar granule are absent. Sporocysts are ellipsoidal, measuring 12 x 9.3 (10.7-12.8x8-10) micron meter. Sporocyst residuum is present as a granulated compact mass. Sporocysts lack a Stieda body. Sporozoites are banana-shaped, laying head to tail in the sporocysts, each with one spherical refractile globule. (author)

  1. Two new species of Eimeria (Apicomplexa, Eimeriidae) from tree skinks, Prasinohaema spp. (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 1991 and June 1992, feces from 4 species of tree skinks, Prasinohaema spp. from Papua New Guinea, were collected and examined for coccidia. Two species, P. flavipes and P. prehensicauda were found to harbor eimerians which are described as new. Oocysts of Eimeria krausi sp. nov. from P. flavipes were ellipsoidal to subspheroidal with a smooth bilayered wall and measured (L × W) 19.2 × 16.9 μm, with a length/width (L/W) ratio of 1.1. Micropyle and oocyst residuum were absent but a fragmented polar granule was present. Sporocysts were ellipsoidal, 9.7 × 6.7 μm, L/W of 1.5. Stieda, subStieda and paraStieda bodies were absent. The sporocyst residuum was composed of many small granules in a compact mass between sporozoites. The sporozoites were sausage-shaped, 11.7 × 2.7 μm, in situ, with an ellipsoidal posterior refractile body and a spheroidal anterior refractile body. Oocysts of Eimeria greeri sp. nov. from P. prehensicauda were ellipsoidal with a smooth bilayered wall, (L × W) 23.0 × 18.3 μm, with a L/W of 1.3. Micropyle and oocyst residuum were absent but a fragmented polar granule was present. Sporocysts were ellipsoidal, 9.7 × 8.4 μm, with a L/W of 1.2. Stieda, subStieda and paraStieda bodies were absent. The sporocyst residuum was composed of many large granules in a compact mass between sporozoites. The sporozoites were sausage-shaped, with an ellipsoidal posterior refractile body and a spheroidal anterior refractile body. We document here the first report of coccidia from skinks of the genus Prasinohaema.

  2. Effects of probiotics and application methods on performance and response of broiler chickens to an Eimeria challenge.

    Science.gov (United States)

    Ritzi, Miranda M; Abdelrahman, Wael; Mohnl, Michaela; Dalloul, Rami A

    2014-11-01

    Coccidiosis is an inherent risk in the commercial broiler industry and inflicts devastating economic losses to poultry operations. Probiotics may provide a potential alternative to the prophylactic use of anticoccidials in commercial production. This study evaluated the effects of probiotic applications (feed and water) on bird performance and resistance to a mixed Eimeria infection in commercial broilers. On day of hatch, 1,008 commercial male broilers (Cobb 500) were assigned to 1 of 6 treatments (8 replicate floor pens; 21 birds/pen), including noninfected negative control (NEG), Eimeria-infected positive control (POS), anticoccidial control (0.01% salinomycin, SAL), intermittent high-dose water-applied probiotic (WPI), continuous low-dose water-applied probiotic (WPC), and feed-supplemented probiotic (FSP). On d 15, all birds except those in NEG were challenged with a mixed inoculum of Eimeria acervulina, Eimeria maxima, and Eimeria tenella. Measurements were taken on d 7, 15, 21, 28, 35, and 42. Fecal samples were collected from d 20 to 24 for oocyst counts, and lesion scores were evaluated on d 21. Data were analyzed using the Fit Model platform in JMP Pro 10.0 (SAS Institute Inc.). Differences in experimental treatments were tested using Tukey's honestly significant difference following ANOVA with significance reported at P ≤ 0.05. Overall, NEG birds outperformed all other groups. For performance, the probiotic groups were comparable with the SAL-treated birds, except during the 6 d immediately following the Eimeria species challenge, where the SAL birds exhibited better performance. The WPC birds had lower duodenal and jejunal lesion scores, indicating a healthier intestine and enhanced resistance to Eimeria species compared with POS. Birds in the WPI treatment shed fewer oocysts in the feces, although this was not a trend for all of the probiotic treatment groups. The results of this study suggest probiotic supplementation without anticoccidials can

  3. Protective Efficacy of Coccidial Common Antigen Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) against Challenge with Three Eimeria Species

    Science.gov (United States)

    Tian, Lu; Li, Wenyu; Huang, Xinmei; Tian, Di; Liu, Jianhua; Yang, Xinchao; Liu, Lianrui; Yan, Ruofeng; Xu, Lixin; Li, Xiangrui; Song, Xiaokai

    2017-01-01

    Coccidiosis is an intestinal disorder of poultry and often caused by simultaneous infections of several Eimeria species. GAPDH is one of the immunogenic common antigens among Eimeria tenella, E. acervulina, and E. maxima identified in our previous study. The present study was performed to further evaluate its immunogenicity and protective efficacy. The genes of GAPDH cloned from E. acervulina and E. maxima were named as EaGAPDH and EmGAPDH, respectively. The immunogenicity of recombinant proteins of EaGAPDH and EmGAPDH were analyzed by Western blot. The transcription and expression of pVAX-EaGAPDH and pVAX-EmGAPDH in the injected muscles were detected by reverse transcription PCR (RT-PCR) and Western blot, respectively. GAPDH-induced changes of T lymphocytes subpopulation, cytokines production, and antibody were determined using flow cytometry, quantitative real-time PCR (qPCR), and ELISA, respectively. Finally, the protective efficacies of pVAX-EaGAPDH and pVAX-EmGAPDH were evaluated by vaccination and challenge experiments. The results revealed that the recombinant GAPDH proteins reacted with the corresponding chicken antisera. The EaGAPDH genes were successfully transcribed and expressed in the injected muscles. Vaccination with pVAX-EaGAPDH and pVAX-EmGAPDH significantly increased the proportion of CD4+ and CD8+ T lymphocytes, the cytokines productions of IFN-γ, IL-2, IL-4 et al., and IgG antibody levels compared to controls. The vaccination increased the weight gains, decreased the oocyst outputs, alleviate the enteric lesions compared to controls, and induced moderate anti-coccidial index (ACI). In conclusion, the coccidial common antigen of GAPDH induced significant humoral and cellular immune response and effective protection against E. tenella, E. acervulina, E. maxima, and mixed infection of the three Eimeria species. PMID:28769877

  4. Chicken IL-17F: identification and comparative expression analysis in Eimeria-infected chickens.

    Science.gov (United States)

    Kim, Woo H; Jeong, Jipseol; Park, Ae R; Yim, Dongjean; Kim, Yong-Hwan; Kim, Kwang D; Chang, Hong H; Lillehoj, Hyun S; Lee, Byung-Hyung; Min, Wongi

    2012-11-01

    Interleukin-17F (IL-17F) is a proinflammatory cytokine, which plays an important role in gut homeostasis. A full-length chicken IL-17F (chIL-17F) cDNA with a 510-bp coding region was identified from ConA-activated chicken splenic lymphocytes. ChIL-17F shares 53% amino acid sequence identity with the previously described chicken IL-17 (chIL-17A) and 38-43% with mammalian homologues. The locus harboring chIL-17 and chIL-17F displayed inverted order compared to those of mammals. ChIL-17F transcript expression was high in lymphoblast cell line CU205 and at moderate levels in small and large intestines and liver. ChIL-17F and chIL-17 expression profiles were examined by quantitative real-time RT-PCR in mitogen-stimulated splenic lymphocytes and intestinal areas affected by Eimeria maxima and Eimeria tenella infections. Expression levels of chIL-17F, like chIL-17, were elevated in mitogen-activated splenic lymphocytes. ChIL-17F, but not chIL-17, expression was upregulated in intestinal tissues affected by E. maxima and E. tenella infections. Recombinant chIL-17F biological activities were similar to that of chIL-17 in primary chicken embryonic fibroblasts. These results suggest that chIL-17F is a unique member of the IL-17 family of cytokines. Copyright © 2012 Elsevier Ltd. All rights reserved.

  5. The mRNA expression of amino acid and sugar transporters, aminopeptidase, as well as the di- and tri-peptide transporter PepT1 in the intestines of Eimeria infected broiler chickens.

    Science.gov (United States)

    Miska, K B; Fetterer, R H

    2017-02-01

    Coccidiosis in chickens is caused by infection of gut epithelial cells with protozoan parasites of the genus Eimeria This disease causes losses to the poultry industry since infected birds fail to gain weight as rapidly as non-infected birds and efficiency of feed conversion is compromised. For the present study the effect of Eimeria on expression of components of amino acid and sugar uptake mechanisms was determined. Broiler chicks were infected with Eimeria maxima, which infects the jejunum; Eimeria acervulina, which infects the duodenum; or Eimeria tenella, which infects the ceca. Sections of the jejunum, duodenum, and ceca (depending on species of Eimeria) were taken at several time points between d zero and 14 post infection (PI) for mRNA expression analysis. Genes examined included one digestive enzyme, 7 peptide and amino acid transporters located on the brush border, 8 transporters located at the basolateral surface of the gut epithelium, and 5 sugar transporters. All 3 Eimeria species examined caused decrease in expression of brush border transporters particularly at d 5 to 7 PI, which corresponds to the time when pathology is greatest. The same pattern was seen in expression of sugar transporters. However, the expression of basolateral transporters differed among species. Eimeria tenella infection resulted in decreased expression of all basolateral transporters, while E. maxima infection caused increased expression of 2 genes and slight decrease in expression of the remaining 5 genes. Infection with E. acervulina resulted in increased expression at the height of infection of all but one basolateral transporter. In conclusion, Eimeria infection causes a general decrease in gene expression of sugar transporter and brush border AATs at the height of infection. However the expression of basolateral transporters is increased in E. maxima and E. acervulina infected birds. It is possible that decreased expression of brush border transporters in combination with

  6. Plasmodium sporozoites trickle out of the injection site.

    Science.gov (United States)

    Yamauchi, Lucy Megumi; Coppi, Alida; Snounou, Georges; Sinnis, Photini

    2007-05-01

    Plasmodium sporozoites make a remarkable journey from the skin, where they are deposited by an infected Anopheline mosquito, to the liver, where they invade hepatocytes and develop into exoerythrocytic stages. Although much work has been done to elucidate the molecular mechanisms by which sporozoites invade hepatocytes, little is known about the interactions between host and parasite before the sporozoite enters the blood circulation. It has always been assumed that sporozoites rapidly exit the injection site, making their interactions with the host at this site, brief and difficult to study. Using quantitative PCR, we determined the kinetics with which sporozoites leave the injection site and arrive in the liver and found that the majority of infective sporozoites remain in the skin for hours. We then performed sub-inoculation experiments which confirmed these findings and showed that the pattern of sporozoite exit from the injection site resembles a slow trickle. Last, we found that drainage of approximately 20% of the sporozoite inoculum to the lymphatics is associated with a significant enlargement of the draining lymph node, a response not observed after intravenous inoculation. These findings indicate that there is ample time for host and parasite to interact at the inoculation site and are of relevance to the pre-erythrocytic stage malaria vaccine effort.

  7. A review of Eimeria infections in horses and other equids

    Science.gov (United States)

    There is considerable confusion concerning validity of Eimeria species in equids, and endogenous developmental stages and pathogenicity of equid Eimeria. This paper summarizes worldwide information on history, structure, life cycle, pathogenicity, prevalence, epidemiology, and diagnosis of Eimeria i...

  8. Immunomodulatory properties of Alternanthera tenella Colla aqueous extracts in mice

    Directory of Open Access Journals (Sweden)

    R.N.M. Guerra

    2003-09-01

    Full Text Available Plants from the genus Alternanthera are thought to possess antimicrobial and antiviral properties. In Brazilian folk medicine, the aqueous extract of A. tenella Colla is used for its anti-inflammatory activity. The present study investigated the immunomodulatory property of A. tenella extract by evaluating the antibody production in male albino Swiss mice weighing 20-25 g (10 per group. The animals received standard laboratory diet and water ad libitum. The effect of A. tenella extract (5 and 50 mg/kg, ip was evaluated in mice immunized with sheep red blood cells (SRBC 10%, ip as T-dependent antigen, or in mice stimulated with mitogens (10 µg, Escherichia coli lipopolysaccharide, LPS, ip. The same doses (5 and 50 mg/kg, ip of A. tenella extract were also tested for antitumor activity, using the Ehrlich ascites carcinoma as model. The results showed that 50 mg/kg A. tenella extract ip significantly enhanced IgM (64% and IgG2a (50% antibody production in mice treated with LPS mitogen. The same dose had no effect on IgM-specific response, whereas the 5 mg/kg treatment caused a statiscally significant reduction of anti-SRBC IgM-specific antibodies (82%. The aqueous extract of A. tenella (50 mg/kg increased the life span (from 16 ± 1 to 25 ± 1 days and decreased the number of viable tumor cells (59% in mice with Ehrlich ascites carcinoma. The present findings are significant for the development of alternative, inexpensive and perhaps even safer strategies for cancer treatment.

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

    International Nuclear Information System (INIS)

    Jenkins, M.C.; Augustine, P.C.; Barta, J.R.; Castle, M.D.; Danforth, H.D.

    1991-01-01

    Sporulated oocysts of the protozoan Eimeria acervulina were subjected to 0, 10, 15, 20, or 30 krad of X-irradiation and inoculated into susceptible outbred chickens to determine if radioattenuated coccidia could induce protection against parasite challenge. Irradiation treatment had an appreciable dose-dependent effect on parasite development. Insignificant numbers of oocysts were produced by chickens inoculated with parasites that had been exposed to greater than 10 krad X-irradiation. Sporozoites exposed to 15 or 20 krad irradiation conferred significant protection against the appearance of intestinal lesions after parasite challenge. Sporozoites subjected to the highest dose level (30 krad) did not produce any significant level of protection. To investigate this phenomenon further and assess intracellular parasite development, susceptible outbred strains of chickens were administered either nonirradiated (0 krad) oocysts or oocysts that were exposed to an optimal dose (15 krad) or a high dose (30 krad) of X-irradiation. Immunofluorescence staining of tissue sections from each treatment group at various intervals after the initial administration of irradiated parasites indicated that sporozoites exposed to 15 krad irradiation were as capable of invading the host intestinal epithelium as nonirradiated sporozoites. However, at 48, 60, 72, and 96 hr, there was a marked reduction in merogonic development in groups receiving irradiated sporozoites compared to those inoculated with nonirradiated parasites. The latter parasites underwent profuse merogonic development; in contrast, irradiated parasites demonstrated little (15 krad) or no (30 krad) merogonic development. These results suggest that induction of a protective immune response occurs during a critical period early in intracellular development of E. acervulina

  10. A NEW SPECIES OF EIMERIA (APICOMPLEXA: EIMERIIDAE) FROM THE EASTERN PIPISTRELLE, PERIMYOTIS SUBFLAVUS (CHIROPTERA: VESPERTILIONIDAE), IN ARKANSAS

    Science.gov (United States)

    McAllister, Chris T.; Burt, Scott; Seville, R. Scott; Robison, Henry W.

    2011-01-01

    During November 2009 and March 2010, 20 adult eastern pipistrelles, Perimyotis (=Pipistrellus) subflavus were collected from Polk County, Arkansas, and their feces examined for coccidian parasites. Two (10%) of the bats were found to be passing oocysts of an undescribed species of Eimeria. Oocysts of Eimeria heidti n. sp. were ovoidal to ellipsoidal, 26.1 × 20.5 (23-31 × 18-23) μm, with a bilayered wall, externally rough, internally smooth, and with a shape index of 1.3. Micropyle and oocyst residuum were absent, but a subspherical polar granule was often present. Sporocysts were ovoidal, 13.0 × 8.8 (11-15 × 7-13) μm, the shape index was 1.6, a Stieda body was present and sub-Stieda and para-Stieda bodies were absent. A sporocyst residuum consisting of multiple globules dispersed along the perimeter of the sporocyst and between the sporozoites were present, sporozoites were elongate, with a subspherical anterior refractile body and elongate posterior refractile body; a nucleus not discernable. This is the second coccidian reported from this host and the fourth instance of a coccidian species reported from an Arkansas bat. PMID:21506799

  11. Expression of digestive enzymes and nutrient transporters in Eimeria-challenged broilers.

    Science.gov (United States)

    Su, S; Miska, K B; Fetterer, R H; Jenkins, M C; Wong, E A

    2015-03-01

    Avian coccidiosis is a disease caused by the intestinal protozoa Eimeria. The site of invasion and lesions in the intestine is species-specific, for example E. acervulina affects the duodenum, E. maxima the jejunum, and E. tenella the ceca. Lesions in the intestinal mucosa cause reduced feed efficiency and body weight gain. The growth reduction may be due to changes in expression of digestive enzymes and nutrient transporters in the intestine. The objective of this study was to compare the expression of digestive enzymes, nutrient transporters and an antimicrobial peptide in broilers challenged with either E. acervulina, E. maxima or E. tenella. The genes examined included digestive enzymes (APN and SI), peptide and amino acid transporters (PepT1, ASCT1, b(0,+)AT/rBAT, B(0)AT, CAT1, CAT2, EAAT3, LAT1, y(+)LAT1 and y(+)LAT2), sugar transporters (GLUT1, GLUT2, GLUT5 and SGLT1), zinc transporter (ZnT1) and an antimicrobial peptide (LEAP2). Duodenum, jejunum, ileum and ceca were collected 7 days post challenge. E. acervulina challenge resulted in downregulation of various nutrient transporters or LEAP2 in the duodenum and ceca, but not the jejunum or ileum. E. maxima challenge produced both downregulation and upregulation of nutrient transporters and LEAP2 in all three segments of the small intestine and ceca. E. tenella challenge resulted in the downregulation and upregulation of nutrient transporters and LEAP2 in the jejunum, ileum and ceca, but not the duodenum. At the respective target tissue, E. acervulina, E. maxima and E. tenella infection caused common downregulation of APN, b(0,+)AT, rBAT, EAAT3, SI, GLUT2, GLUT5, ZnT1 and LEAP2. The downregulation of nutrient transporters would result in a decrease in the efficiency of protein and polysaccharide digestion and uptake, which may partially explain the weight loss. The downregulation of nutrient transporters may also be a cellular response to reduced expression of the host defense protein LEAP2, which would

  12. In vitro infectivity of irradiated Plasmodium berghei sporozoites to cultured hepatoma cells

    International Nuclear Information System (INIS)

    Sigler, C.I.; Leland, P.; Hollingdale, M.R.

    1984-01-01

    The invasion of gamma-irradiated Plasmodium berghei sporozoites into cultured hepatoma cells and their transformation into trophozoites was similar to invasion and transformation of non-irradiated sporozoites. However, trophozoites from irradiated sporozoites did not further develop into schizonts, but persisted within the cells for up to 3 days. Sporozoite surface protective antigen was present in trophozoites from irradiated and non-irradiated sporozoites, suggesting that hepatocyte antigen processing may contribute to the induction of anti-malarial immunity

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

    International Nuclear Information System (INIS)

    Singh, J.; Gill, B.S.

    1975-01-01

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

  14. Eimeria species (Apicomplexa: Eimeriidae of podocnemis expansa (Schweigger and geochelone denticulata (LINN. from Amazonian Brazil (Reptilia: Chelonia

    Directory of Open Access Journals (Sweden)

    Ralph Lainson

    1990-12-01

    Full Text Available Eimeria lagunculata, Eimeria mammiformis and Eimeria podocnemis n. spp., are described from the faeces of the fresh-water turtle Podocnemis expansa, in Pará State, north Brasil. Oocysts of E. lagunculata are ellipsoidal, 19.2 x 12.8 (17.0-20.7 x 11.8-14.1 mum, shape-index (= length/ width 1.5 (1.4-1.7. Oocyst wall about 0.5-0.7 mum thick, with a prominent stopper-like micropyle at one pole. No oocyst residuum and no polar body. Sporocysts elongate ellipsoidal, 11.0 x 5.4 (10.4-11.8 x 5.2-6.0 mum, shape-index 2.0 (1.8-2.1: no Stieda body. A compact, ellipsoidal sporocyst residuum lies between the two sporozoites, which possess a posterior and an anterior refractile body. Oocysts of E. mammiformis broadly ellipsoidal, 30.0 x 19.4 (23.0-37.0 x 16.3-21.5 mum, shape-index 1.5 (1.1-1.9. Oocyst wall about 0.7 mum thick, with a prominent micropyle: no oocyst residuum and rarely a single polar body. Sporocysts ellipsoidal, 15.3 x 7.9 (14.8-17.0 x 7.4-9.6 mum, shape-index 2.0 (1.8-2.2, with a tiny Stieda body. Sporocyst residuum bulky, ellipsoidal: sporozoites with two conspicuous refractile bodies. E. podocnemis has broadly ellipsoidal oocysts, 17.0 x 12.8 (14.8-19.2 x 11.8-14.1 mum, shape-index 1.3 (1.1-1.4. Oocyst wall about 0.5-0.7 mum thick, with no micropyle. No oocyst residuum, but always a single polar body. Sporocysts ellipsoidal, 9.7 x 5.2 (8.9-10.4 x 4.4-6.0 mum, shape-index 1.9 (1.6-2.0, with no Stieda body. Sporocyst residuum bulky, ellipsoidal: sporocysts with 2 refractile bodies. Eimeria carinii n. sp., is recorded from the tortoise Geochelone denticulata, also from Pará. Oocyst wall about 1.2 mum thicl. No micropyle. Oocyst residuum limited to a number (about 10-20 of scattered granules: no polar body. Sporocysts broadly ellipsoidal, and with no Stieda body: they measure 8,8 x 7.3 (8.0-9.0 x 7.0-7.5 mum, shape-index 1.2 (1.1-1.3. Sporocyst residuum bulky, spherical to ellipsoidal: sporozoites possess both posterior and anterior

  15. Two new species of Eimeria (Apicomplexa: Eimeriidae) from the mountain beaver, Aplodontia rufa (Rodentia: Aplodontiidae), from Oregon.

    Science.gov (United States)

    McAllister, Chris T; Duszynski, Donald W; McKown, Richard D

    2013-06-01

    Two mountain beavers, Aplodontia rufa , were collected in Lincoln County, Oregon, and examined for coccidia. Both were infected with 2 new species of Eimeria. Oocysts of Eimeria chitkoae n. sp. were ellipsoidal with a bilayered wall and measured (L × W) 24.5 × 20.2 μm, with a shape index (SI) of 1.2. Both micropyle and oocyst residuum were absent, but a polar granule of several fragments was present. Sporocysts were ovoidal, 12.5 × 7.9 μm, SI was 1.6. Stieda and substieda bodies were present, but a parastieda body was absent; a sporocyst residuum was present, composed of a cluster of moderately coarse granules with many scattered fine granules. Stout sporozoites were 14.7 × 2.9 μm in situ, with spheroidal anterior and posterior refractile bodies. Oocysts of Eimeria lewisi n. sp. were ovoidal, with a smooth single-layered wall, and measured 13.7 × 7.8 μm, SI was 1.7. A micropyle and oocyst residuum were absent, but 1-2 polar granule(s) were present. Sporocysts were 6.6 × 4.2 μm, with SI of 1.6. A Stieda body was present, but substieda and parastieda bodies were absent; a sporocyst residuum was present, composed of a small cluster of several granules. Sporozoites were granular, 8.2 × 1.8 μm in situ, with a posterior refractile body. These are the first coccidians reported from the mountain beaver.

  16. Molecular characterisation and the protective immunity evaluation of Eimeria maxima surface antigen gene.

    Science.gov (United States)

    Liu, Tingqi; Huang, Jingwei; Li, Yanlin; Ehsan, Muhammad; Wang, Shuai; Zhou, Zhouyang; Song, Xiaokai; Yan, Ruofeng; Xu, Lixin; Li, Xiangrui

    2018-05-30

    Coccidiosis is recognised as a major parasitic disease in chickens. Eimeria maxima is considered as a highly immunoprotective species within the Eimeria spp. family that infects chickens. In the present research, the surface antigen gene of E. maxima (EmSAG) was cloned, and the ability of EmSAG to stimulate protection against E. maxima was evaluated. Prokaryotic and eukaryotic plasmids expressing EmSAG were constructed. The EmSAG transcription and expression in vivo was performed based on the RT-PCR and immunoblot analysis. The expression of EmSAG in sporozoites and merozoites was detected through immunofluorescence analyses. The immune protection was assessed based on challenge experiments. Flow cytometry assays were used to determine the T cell subpopulations. The serum antibody and cytokine levels were evaluated by ELISA. The open reading frame (ORF) of EmSAG gene contained 645 bp encoding 214 amino acid residues. The immunoblot and RT-PCR analyses indicated that the EmSAG gene were transcribed and expressed in vivo. The EmSAG proteins were expressed in sporozoite and merozoite stages of E. maxima by the immunofluorescence assay. Challenge experiments showed that both pVAX1-SAG and the recombinant EmSAG (rEmSAG) proteins were successful in alleviating jejunal lesions, decreasing loss of body weight and the oocyst ratio. Additionally, these experiments possessed anticoccidial indices (ACI) of more than 170. Higher percentages of CD4 + and CD8 + T cells were detected in both EmSAG-inoculated birds than those of the negative control groups (P maxima.

  17. Experimental vaccination of chicks with Plasmodium gallinaceum sporozoites. I. Circumsporozoite proteins are expressed by sporozoites recovered from both salivary glands and midguts of mosquitoes

    International Nuclear Information System (INIS)

    Daher, V.R.; Krettli, A.U.

    1987-01-01

    Immunogenicity of Plasmodium gallinaceum sporozoites for chicks and their in vitro reactivity with normal and specific immune sera were studied. Two sporozoite populations recovered from experimentally infected Aedes fluviatilis were used: sporozoites from salivary glands and sporozoites from midgut oocysts. Populations seven to nine days old of sporozoites recovered from salivary glands were infective for all chicks until the chicks were three weeks old; however, sporozoites recovered from midguts containing oocysts infected these chicks only if isolated on days 8-9, but not on day 7 after the mosquitoes' infective blood meal. Infectivity of the sporozoites was lost after exposure to ultraviolet (UV) light (30 min) or X-rays (13 krad). Inactivated sporozoites from both sources proved highly immunogenic to chicks that were immunized by several intravenous or intramuscular injections. These parasites elicited a strong humoral immune response in the chicks, as measured by the circumsporozoite precipitation (CSP) reaction. The levels of the CSP antibodies were similar with sporozoites from both sources, there being no detectable differences in the percentage of reactive sporozoites or the intensity of the CSP reaction with sera containing antibodies to either sporozoites from salivary glands or sporozoites from oocysts. These results provide the first evidence that avian malaria sporozoites express the circumsporozoite protein that has been extensively characterized in mammalian malaria (rodent, simian, human sporozoites). Furthermore, we observed that the yields of sporozoites obtained from mosquito midguts, on days 8 and 9 of the P. gallinaceum infection, were at least twice as great as those obtained by salivary gland dissection, even 20 days after a blood meal

  18. Complete mitochondrial genome sequences from five Eimeria species (Apicomplexa; Coccidia; Eimeriidae) infecting domestic turkeys

    OpenAIRE

    Ogedengbe, Mosun E; El-Sherry, Shiem; Whale, Julia; Barta, John R

    2014-01-01

    Background Clinical and subclinical coccidiosis is cosmopolitan and inflicts significant losses to the poultry industry globally. Seven named Eimeria species are responsible for coccidiosis in turkeys: Eimeria dispersa; Eimeria meleagrimitis; Eimeria gallopavonis; Eimeria meleagridis; Eimeria adenoeides; Eimeria innocua; and, Eimeria subrotunda. Although attempts have been made to characterize these parasites molecularly at the nuclear 18S rDNA and ITS loci, the maternally-derived and mitotic...

  19. 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.

  20. An alternative to antibiotic-based drugs in feed for enhancing performance of broilers grown on Eimeria spp.-infected litter.

    Science.gov (United States)

    Stanley, V G; Gray, C; Daley, M; Krueger, W F; Sefton, A E

    2004-01-01

    Three trials were conducted to evaluate the effects of lasalocid, an anticoccidial feed additive (90.7 kg/ton); bacitracin, a growth-promoter (50 g/ton); and yeast culture residue (YCR) (1 kg/ton) on the performance of broiler chicks reared to 42 d of age on recycled litter. Recycled litter consisted of pine wood shavings containing droppings from chicks infected with 3 select strains of coccidia (Eimeria tenella, Eimeria maxima, and Eimeria acervulina). Response variables (BW, intestinal tract and litter coliform counts, cecal and liver relative weights, and litter moisture content) were recorded biweekly. Mean BW of chicks fed the diet supplemented with YCR was higher than that of the controls (P litter aged and moisture content increased. The mean intestinal coliform population from YCR-treated chicks was lower (P litter coliform counts increased with increased use of the litter. Cecal and liver relative weights calculated from the chicks in trial 3 showed that only the liver was significantly affected by treatments. YCR appeared to be a viable alternative to bacitracin and lasalocid medication in enhancing growth of broiler chicks reared on recycled litter.

  1. 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

  2. A new species of Eimeria Schneider, 1875 (Apicomplexa: Eimeriidae) from the Solomon ground skink, Sphenomorphus solomonis (Boulenger) (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 1990 and November 1991, 19 Sphenomorphus spp. skinks, including nine S. jobiense, three S. simus, and seven Solomon ground skinks, S. solomonis (Boulenger), were collected from Madang and Morobe Provinces, Papua New Guinea (PNG), and examined for coccidia. A single S. solomonis was found to be infected with a new species of Eimeria Schneider, 1875. Oöcysts of Eimeria perkinsae n. sp. are ellipsoidal with a smooth, colourless, bi-layered wall, measure 18.6 × 14.7 μm, and have a length/width (L/W) ratio of 1.3; both micropyle and oöcyst residuum are absent, but a fragmented polar granule is present. Sporocysts are ovoidal, 8.9 × 6.4 μm, L/W 1.4; neither Stieda, sub-Stieda or para-Stieda bodies are present; a sporocyst residuum consisted of a loose cluster of granules dispersed between sporozoites. Sporozoites are comma-shaped with spheroidal anterior and posterior refractile bodies. This represents the first report of coccidia from this skink genus.

  3. Identification of a surface antigen on Theileria parva sporozoites by monoclonal antibody.

    OpenAIRE

    Dobbelaere, D A; Shapiro, S Z; Webster, P

    1985-01-01

    A mouse monoclonal antibody (mAbD1) that neutralizes sporozoites of different stocks of the protozoan parasite Theileria parva has been used to localize and identify a sporozoite antigen. Protein A-colloidal gold was used to localize bound mAbD1 in immunoelectron microscopic studies. mAbD1 bound to sporozoite antigen, which was evenly spread over the surface of all sporozoites. Immune complexes were obtained by incubation of sporozoite suspensions with mAbD1 followed by Zwittergent 3-14 extra...

  4. Survival and antigenic profile of irradiated malarial sporozoites in infected liver cells

    International Nuclear Information System (INIS)

    Suhrbier, A.; Winger, L.A.; Castellano, E.; Sinden, R.E.

    1990-01-01

    Exoerythrocytic (EE) stages of Plasmodium berghei derived from irradiated sporozoites were cultured in vitro in HepG2 cells. They synthesized several antigens, predominantly but not exclusively those expressed by normal early erythrocytic schizonts. After invasion, over half the intracellular sporozoites, both normal and irradiated, appeared to die. After 24 h, in marked contrast to the normal parasites, EE parasites derived from irradiated sporozoites continued to break open, shedding their antigens into the cytoplasm of the infected host cells. Increasing radiation dosage, which has previously been shown to reduce the ability of irradiated sporozoites to protect animals, correlated with reduced de novo antigen synthesis by EE parasites derived from irradiated sporozoites

  5. Improvement of Theileria parva sporozoite vaccine against East ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Mobile Nav Footer Links ... Improvement of Theileria parva sporozoite vaccine against East Coast fever ... Mortality rates approach 90% in susceptible cattle, and animals that recover are long-term Theileria parva ... Global Alliance for Livestock Veterinary Medicines (GALVmed) for technology distribution to endemic regions.

  6. Antibody and B cell responses to Plasmodium sporozoites

    Directory of Open Access Journals (Sweden)

    Johanna N Dups

    2014-11-01

    Full Text Available Antibodies are capable of blocking infection of the liver by Plasmodium sporozoites. Accordingly the induction of anti-sporozoite antibodies is a major aim of various vaccine approaches to malaria. In recent years our knowledge of the specificity and quantities of antibodies required for protection has been greatly expanded by clinical trials of various whole sporozoite and subunit vaccines. Moreover, the development of humanized mouse models and transgenic parasites have also aided our ability to assess the specificity of antibodies and their ability to block infection. Nonetheless, considerable gaps remain in our knowledge - in particular in understanding what antigens are recognized by infection blocking antibodies and in knowing how we can induce robust, long-lived antibody responses. Maintaining high levels of circulating antibodies is likely to be of primary importance, as antibodies must block infection in the short time it takes for sporozoites to reach the liver from the skin. It is clear that a better understanding of the development of protective B cell-mediated immunity will aid the development and refinement of malaria vaccines.

  7. Differences in susceptibility among mouse strains to infection with Plasmodium berghei (ANKA clone) sporozoites and its relationship to protection by gamma-irradiated sporozoites

    International Nuclear Information System (INIS)

    Jaffe, R.I.; Lowell, G.H.; Gordon, D.M.

    1990-01-01

    Three inbred mouse strains, C57BL/6 (H-2b), A/J (H-2a), and BALB/c (H-2d), and 1 outbred strain, CD-1, demonstrated differences in susceptibility to iv challenge with the ANKA clone of Plasmodium berghei. Mice were challenged with 100, 1,000, or 10,000 sporozoites, then evaluated daily beginning on day 4 for patency. CD-1 mice were further evaluated at challenge doses of 12,500, 25,000, and 50,000 sporozoites. C57BL/6 mice were the easiest to infect, with 90% becoming infected with 100 sporozoites. The outbred strain CD-1 was the most difficult to infect, requiring a challenge dose of 25,000 sporozoites/mouse in order to achieve a 100% infection rate. Mouse strains also demonstrated differences in their ability to be protected by intravenous immunization with gamma-irradiated sporozoites. A/J mice needed a minimum of 3 doses of irradiated sporozoites for protection against a challenge with 10,000 sporozoites. In contrast, BALB/c mice immunized with a single dose of 1,000 irradiated sporozoites are protected against a 10,000 sporozoite challenge. These data suggest that both infectivity and protection are genetically restricted and that susceptibility to infection may be inversely related to protection

  8. Stable transfection of Eimeria intestinalis and investigation of its life cycle, reproduction and immunogenicity

    Directory of Open Access Journals (Sweden)

    Tuanyuan eShi

    2016-05-01

    Full Text Available Rabbit coccidiosis, caused by infection of Eimeria spp. is one of the most severe parasitic diseases in rabbits. E. intestinalis is one of the most immunogenic species in rabbit coccidia. Due to the lack of genomic information and unsuccessful in vitro cultivation, genetic manipulation of rabbit coccidia lagged behind other apicomplexan parasites. Using regulatory sequences from E. tenella, we obtained a transgenic line of E. intestinalis expressing yellow fluorescent protein (YFP. YFP was continuously expressed throughout the whole life cycle. Morphological features of E. intestinalis in the different developmental stages were dynamically observed with the transgenic line. Some important features in the endogenous development stages were observed. Trophozoites were found as early as 4 h post inoculation. Two-types of schizonts and merozoites were observed in first three of the four schizogonies. Beside jejunum and ileum, gametogony stage and oocysts were also found in the duodenum and vermiform appendix. In addition, the transgenic strain was highly immunogenic but less pathogenic than the wild type. Considering the high immunogenicity of E. intestinalis and amenability to transfection with foreign genes, transgenic E. intestinalis could be a promising oral eukaryotic vaccine vector.

  9. Eimeria infections in goats in Southern Portugal

    Directory of Open Access Journals (Sweden)

    Liliana Machado Ribeiro da Silva

    Full Text Available 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.

  10. Eimeria collieie n. sp. (Apicomplexa:Eimeriidae) from the western long-necked turtle (Chelodina colliei).

    Science.gov (United States)

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

    2015-07-01

    A new species, Eimeria collieie n. sp., is described from the western long-necked turtle (Chelodina colliei). Sporulated oocysts (n = 35) are spherical to subspherical, with colourless single layer oocyst wall, 0.6 ± 0.2 (0.4-0.7) µm thick. Oocyst with elongated ellipsoid sporocysts. Oocyst length, 29.8 ± 0.4 (28.2-31.0) µm; oocyst width, 29.4 ± 0.3 (28.0-30.8) µm; oocyst length/width (L/W) ratio, 1.0 ± 0.03 (1.0-1.05). Micropyle, oocyst residuum and polar granule were absent. Sporocysts with sporocyst residuum and 2 sporozoites. Sporocyst length, 21.6 ± 0.4 (21.2-22.0) µm; sporocyst width, 6.0 ± 0.3 (5.7-6.3) µm; sporocyst L/W ratio, 3.6 ± 0.2 (3.4-3.8). Stieda, parastieda and substieda bodies were absent. Sporozoite length, 14.0 ± 0.2 (13.8-14.2) µm; sporozoite width, 2.6 ± 0.2 (2.4-2.8) µm; sporozoite L/W ratio, 5.46 ± 0.10 (5.4-5.6). Molecular analysis was conducted at three loci: the 18S and 28S ribosomal RNA (rRNA), and the mitochondrial cytochrome oxidase gene (COI). At the 18S rRNA locus, E. collieie n. sp. shared 96.4% and 98.3% genetic similarity to E. ranae (GenBank accession number: EU717219) and E. arnyi (AY613853) respectively. At the 28S rRNA locus, E. collieie n. sp. shared 91.6% genetic similarity to E. papillata (GenBank accession number: GU593706) and phylogenetic analysis at this locus placed E. collieie n. sp. in aseparateclade. At the COI locus, E. collieie n. sp. shared 92.7% genetic similarity to Eimeria setonicis (GenBankaccession number: KF225638) from a quokka (Setonix brachyurus) in Western Australia. Reptile-derived sequences were not available for the 28S rRNA and the COI loci. Based on morphological and molecular data, this isolate is a new species of coccidian parasite that, to date, has only been found in western long-necked turtles. Crown Copyright © 2015. Published by Elsevier Inc. All rights reserved.

  11. Eimeria pavoaegyptica sp. nov. (Apicomplexa: Eimeriidae) in faeces of Indian peacocks, Pavo cristatus Linnaeus, 1758 (Galliformes: Phasianidae) from Egypt.

    Science.gov (United States)

    El-Shahawy, Ismail Saad

    2010-12-01

    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) microm 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 ~approximately1.7 microm 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) microm; their average shape-index is 2.5 microm 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 microm and each has two different-sized refractile bodies.

  12. 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.

  13. A new species of Eimeria Schneider, 1875 (Apicomplexa: Eimeriidae) from Carlia spp. (Sauria: Scincidae) from Papua New Guinea

    Science.gov (United States)

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

    2013-01-01

    A new species of Eimeria Schneider, 1875 from rainbow skinks, Carlia ailanpalai Zug and Carlia eothen Zug is described from specimens collected in Papua New Guinea (PNG). Oöcysts of Eimeria zugi n. sp. from one of one (100%) C. eothen are ellipsoidal to cylindroidal, with a smooth, colourless, bi-layered wall, measure 25.1 × 15.5 μm and have a length/width ratio of 1.6. The micropyle and the oöcyst residuum are absent, but a polar granule is present. The sporocysts are ovoidal to ellipsoidal and 10.3 × 7.1 μm in size and do not contain Stieda, sub-Stieda or para-Stieda bodies; and the sporocyst residuum is composed of a compact mass of large globules. The sporozoites are elongate, 12.8 × 2.9 μm in size, and contain anterior and posterior refractile bodies with a nucleus between them. This is the ninth species of coccidium described from skinks from PNG, and the new species described herein is apparently endemic to the skink genus Carlia (Gray).

  14. First field trial of an immunoradiometric assay for the detection of malaria sporozoites in mosquitoes

    International Nuclear Information System (INIS)

    Collins, F.H.; Zavala, F.; Graves, P.M.; Cochrane, A.H.; Gwadz, R.W.; Akoh, J.; Nussenzweig, R.S.

    1984-01-01

    An immunoradiometric assay (IRMA) using a monoclonal antibody to the major surface protein of Plasmodium falciparum sporozoites was used to assess the P. falciparum sporozoite rate in a West African population of Anopheles gambiae (s.1.). Unlike current dissection techniques, the IRMA could detect sporozoite antigen in dried as well as fresh mosquitoes. In a controlled comparison, the sensitivity of the IRMA was comparable that of the dissection technique. Additionally, the IRMA was species specific and quantitative. Sensitivity of the assay was sufficient to detect sporozoite infections resulting from the development of a single oocyst

  15. Transmission dynamics of Eimeria acervulina in broilers

    NARCIS (Netherlands)

    Velkers, F.C.

    2011-01-01

    Control of the intestinal disease coccidiosis, caused by infections with Eimeria species, is a major challenge, especially for the broiler industry. Effective control strategies require a comprehensive understanding of processes that lead to infection and disease in a population. One of the key

  16. New approaches for morphological diagnosis of bovine Eimeria species: a study on a subtropical organic dairy farm in Brazil.

    Science.gov (United States)

    Florião, Mônica Mateus; Lopes, Bruno do Bomfim; Berto, Bruno Pereira; Lopes, Carlos Wilson Gomes

    2016-03-01

    Bovine eimeriosis or coccidiosis is an intestinal disease caused by Eimeria spp. which is related to gastrointestinal disorders and, in some cases, death. The current work aimed to identify and provide detailed morphological characteristic features of the different Eimeria spp. parasites of crossbred cows of a subtropical organic dairy farm in Brazil, offering tools for the diagnosis of bovine eimeriosis. Eimeria auburnensis, Eimeria bovis, Eimeria bukidnonensis, Eimeria canadensis, Eimeria cylindrica, Eimeria ildefonsoi, and Eimeria zuernii were identified. The application of line regressions and ANOVA provided a means for the identification of these species. Finally, the current work proposes a dichotomous key to assist in the morphologic identification of bovine Eimeria spp. oocysts.

  17. Genetically attenuated P36p-deficient Plasmodium berghei sporozoites confer long-lasting and partial cross-species protection

    NARCIS (Netherlands)

    Douradinha, Bruno; van Dijk, Melissa R.; Ataide, Ricardo; van Gemert, Geert-Jan; Thompson, Joanne; Franetich, Jean-Francois; Mazier, Dominique; Luty, Adrian J. F.; Sauerwein, Robert; Janse, Chris J.; Waters, Andrew P.; Mota, Maria M.

    2007-01-01

    Immunisation with live, radiation-attenuated sporozoites (RAS) or genetically attenuated sporozoites (GAS) of rodent plasmodial parasites protects against subsequent challenge infections. We recently showed that immunisation with Plasinodium berghei GAS that lack the microneme protein P36p protects

  18. Morphological and molecular characterization of Eimeria labbeana-like (Apicomplexa:Eimeriidae) in a domestic pigeon (Columba livia domestica, Gmelin, 1789) in Australia.

    Science.gov (United States)

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

    2016-07-01

    An Eimeria species is described from a domestic pigeon (Columba livia domestica). Sporulated oocysts (n = 35) were subspherical, with a smooth bi-layered oocyst wall (1.0 μm thick). Oocysts measured 20.2 × 16.1 (22.0-18.9 × 15.7-18.9) μm, oocyst length/width (L/W) ratio, 1.38. Oocyst residuum and a polar granule were present. The micropyle was absent. Sporocysts are elongate-ovoid, 13.0 × 6.1 (14.5-12.5 × 5.5-7.0) μm, sporocyst L/W ratio, 2.13 (2.0-2.2), sporocyst residuum was present, composed of numerous granules in a spherical or ovoid mass. Each sporocyst contained 2 banana-shaped sporozoites, 12.3 × 3.5 (11.8-13.0 × 3.3-3.6) μm. A spherical-ellipsoid posterior refractile body was found in the sporozoites. A nucleus was located immediately anterior to the posterior refractile body. Molecular analysis was conducted at three loci; the 18S and 28S ribosomal RNA genes and the mitochondrial cytochrome oxidase gene (COI). At the 18S locus, the new isolate shared 98.0% genetic similarity with three Isospora isolates from Japan from the domestic pigeon (Columba livia domestica). At the 28S locus, it grouped separately and shared 92.4% and 92.5% genetic similarity with Isospora anthochaerae (KF766053) from a red wattlebird (Anthochaera carunculata) from Australia and an Isospora sp. (MS-2003 - AY283845) from a Himalayan grey-headed bullfinch (Pyrrhula erythaca) respectively. At COI locus, this new isolate was in a separate clade and shared 95.6% and 90.0% similarity respectively with Eimeria tiliquae n. sp. from a shingleback skink in Australia and an Eimeria sp. from a common pheasant (Phasianus colchicus) from America. Based on the morphological data, this isolate is most similar to Eimeria labbeana. As no molecular data for E. labbeana is available and previous morphological data is incomplete, we refer to the current isolate as E. labbeana-like. Crown Copyright © 2016. Published by Elsevier Inc. All rights reserved.

  19. Sequence analysis of the PIP5K locus in Eimeria maxima provides further evidence for eimerian genome plasticity and segmental organization.

    Science.gov (United States)

    Song, B K; Pan, M Z; Lau, Y L; Wan, K L

    2014-07-29

    Commercial flocks infected by Eimeria species parasites, including Eimeria maxima, have an increased risk of developing clinical or subclinical coccidiosis; an intestinal enteritis associated with increased mortality rates in poultry. Currently, infection control is largely based on chemotherapy or live vaccines; however, drug resistance is common and vaccines are relatively expensive. The development of new cost-effective intervention measures will benefit from unraveling the complex genetic mechanisms that underlie host-parasite interactions, including the identification and characterization of genes encoding proteins such as phosphatidylinositol 4-phosphate 5-kinase (PIP5K). We previously identified a PIP5K coding sequence within the E. maxima genome. In this study, we analyzed two bacterial artificial chromosome clones presenting a ~145-kb E. maxima (Weybridge strain) genomic region spanning the PIP5K gene locus. Sequence analysis revealed that ~95% of the simple sequence repeats detected were located within regions comparable to the previously described feature-rich segments of the Eimeria tenella genome. Comparative sequence analysis with the orthologous E. maxima (Houghton strain) region revealed a moderate level of conserved synteny. Unique segmental organizations and telomere-like repeats were also observed in both genomes. A number of incomplete transposable elements were detected and further scrutiny of these elements in both orthologous segments revealed interesting nesting events, which may play a role in facilitating genome plasticity in E. maxima. The current analysis provides more detailed information about the genome organization of E. maxima and may help to reveal genotypic differences that are important for expression of traits related to pathogenicity and virulence.

  20. Eimeria minasensis n. sp. (Apicomplexa: Eimeriidae in the Domestic Goat Capra hircus, from Brazil

    Directory of Open Access Journals (Sweden)

    Silva Andréa C

    1998-01-01

    Full Text Available Eimeria minasensis n. sp. is described in the domestic goat Capra hircus from Brazil. Oocysts ellipsoidal are 35 x 24.5 (32-37.7 x 20.9-27.9 mm. Sporocysts elongate-ellipsoid are 15.2 x 9 (12.3-18.4 x 7.8-10.2 mm, with a Stieda body at the narrow end. Oocyst wall smooth and bilayered; outer layer about 1.2 (0.8-1.6 mm and colorless; inner layer about 0.5 (0.4-0.8 mm and dark-brown. Micropyle, a mound-shaped micropylar cap 1,6 x 8,9 (0,8-2 x7-10,2 easily dislodged; one or more oocyst polar granules present. Oocyst residuum absent. Sporocyst residuum present, composed of many scattered granules. Sporozoites elongate, lying lengthwise, "head to tail" in the sporocysts; one or two refractile globules are usually visible. Sporulation time was 120 hr at 27oC, prepatent period, 19 to 20 days and patent period 15 to 25 days. Gamonts, gametes and oocysts present in cecum and colon. Prevalence was 12.8% (6/47 in goats from Minas Gerais, Brazil.

  1. Protective immunity against Eimeria maxima induced by vaccines of Em14-3-3 antigen.

    Science.gov (United States)

    Liu, Tingqi; Huang, Jingwei; Ehsan, Muhammad; Wang, Shuai; Fei, Hong; Zhou, Zhouyang; Song, Xiaokai; Yan, Ruofeng; Xu, Lixin; Li, Xiangrui

    2018-04-15

    Eimeria maxima 14-3-3 (Em14-3-3) open reading frame (ORF) which consisted of 861 bp encoding a protein of 286 amino acids was successfully amplified and sequenced. Subsequently, the Em14-3-3 ORF was subcloned into pET-32a (+) and pVAX1, respectively. RT-PCR and immunoblot analyses confirmed that the target gene was successfully transcribed and expressed in vivo. Immunofluorescence analysis showed that Em14-3-3 was expressed in both the sporozoites and merozoites. The animal experiments demonstrated that both rEm14-3-3 and pVAX1-14-3-3 could clearly alleviate jejunum lesions and body weight loss. The Em14-3-3 vaccines could increase oocyst decrease ratio, as well as produce an anticoccidial index of more than 165. The percentages of CD4 + in both the Em14-3-3 immunized groups were much higher, when compared with those of PBS, pET32a (+), and pVAX1 controls (P maxima. Copyright © 2018 Elsevier B.V. All rights reserved.

  2. 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.

  3. Liver or blood-stage arrest during malaria sporozoite immunization: the later the better?

    NARCIS (Netherlands)

    Nganou Makamdop, C.K.; Sauerwein, R.W.

    2013-01-01

    So far, the best immunization strategies to achieve high levels of protection against malaria are based on whole parasites. Complete sterile protection can be obtained in rodent models after immunization with sporozoites and chemoprophylaxis, or with sporozoites attenuated either genetically or by

  4. Hepatocyte CD81 is required for Plasmodium falciparum and Plasmodium yoelii sporozoite infectivity.

    NARCIS (Netherlands)

    Silvie, O.; Rubinstein, E.; Franetich, J.F.; Prenant, M.; Belnoue, E.; Renia, L.; Hannoun, L.; Eling, W.M.C.; Levy, S.; Boucheix, C.; Mazier, D.

    2003-01-01

    Plasmodium sporozoites are transmitted through the bite of infected mosquitoes and first invade the liver of the mammalian host, as an obligatory step of the life cycle of the malaria parasite. Within hepatocytes, Plasmodium sporozoites reside in a membrane-bound vacuole, where they differentiate

  5. Antisporozoite antibodies in mice immunized with irradiation-attenuated Plasmodium berghei sporozoites

    International Nuclear Information System (INIS)

    Hansen, R.; Silva, S.de; Strickland, G.T.

    1979-01-01

    Sera from NMRI/NIH mice were tested for the presence of IgM and IgG anti-sporozoite antibodies using the indirect fluorescent antibody test (IFAT). Both IgM and IgG antibody titres were related to the number of immunizations with irradiation-attenuated Plasmodium berghei sporozoites, and protection from challenge with subsequent non-attenuated sporozoites correlated with the pre-challenge antibody titre. Sera taken five days following challenge showed marked reductions in antibody titres, except for the group receiving the maximum (four) immunizations. Groups immunized with frozen sporozoites or mosquito tissue antigen developed neither antibodies to sporozoites nor protective immunity; nor did animals infected with parasitized blood. However, sera from mice immunized four times with attenuated sporozoites demonstrated IFA titres to blood-stage antigens. The results showed that both IgM and IgG anti-sporozoite antibodies could be detected in mice immunized with attenuated-sporozoites by IFAT, and that the antibody titres correlated with protective immunity. Cross reaction with blood-stage antigens occurred, but the test should still prove useful. (author)

  6. Proteomic profiling of Plasmodium sporozoite maturation identifies new proteins essential for parasite development and infectivity.

    NARCIS (Netherlands)

    Lasonder, E.; Janse, C.J.; Gemert, G.J.A. van; Mair, G.R.; Vermunt, A.M.W.; Douradinha, B.G.; Noort, V. van; Huynen, M.A.; Luty, A.J.F.; Kroeze, H.; Khan, S.M.; Sauerwein, R.W.; Waters, A.P.; Mann, M.; Stunnenberg, H.G.

    2008-01-01

    Plasmodium falciparum sporozoites that develop and mature inside an Anopheles mosquito initiate a malaria infection in humans. Here we report the first proteomic comparison of different parasite stages from the mosquito -- early and late oocysts containing midgut sporozoites, and the mature,

  7. Plasmodium knowlesi Sporozoite Antigen: Expression by Infectious Recombinant Vaccinia Virus

    Science.gov (United States)

    Smith, Geoffrey L.; Godson, G. Nigel; Nussenzweig, Victor; Nussenzweig, Ruth S.; Barnwell, John; Moss, Bernard

    1984-04-01

    The gene coding for the circumsporozoite antigen of the malaria parasite Plasmodium knowlesi was inserted into the vaccinia virus genome under the control of a defined vaccinia virus promoter. Cells infected with the recombinant virus synthesized polypeptides of 53,000 to 56,000 daltons that reacted with monoclonal antibody against the repeating epitope of the malaria protein. Furthermore, rabbits vaccinated with the recombinant virus produced antibodies that bound specifically to sporozoites. These data provide evidence for expression of a cloned malaria gene in mammalian cells and illustrate the potential of vaccinia virus recombinants as live malaria vaccines.

  8. Proteomic profiling of Plasmodium sporozoite maturation identifies new proteins essential for parasite development and infectivity

    DEFF Research Database (Denmark)

    Lasonder, Edwin; Janse, Chris J; van Gemert, Geert-Jan

    2008-01-01

    Plasmodium falciparum sporozoites that develop and mature inside an Anopheles mosquito initiate a malaria infection in humans. Here we report the first proteomic comparison of different parasite stages from the mosquito -- early and late oocysts containing midgut sporozoites, and the mature...... whose annotation suggest an involvement in sporozoite maturation, motility, infection of the human host and associated metabolic adjustments. Analyses of proteins identified in the P. falciparum sporozoite proteomes by orthologous gene disruption in the rodent malaria parasite, P. berghei, revealed...... three previously uncharacterized Plasmodium proteins that appear to be essential for sporozoite development at distinct points of maturation in the mosquito. This study sheds light on the development and maturation of the malaria parasite in an Anopheles mosquito and also identifies proteins that may...

  9. The trial detection of malaria sporozoit in field-collected mosquito by immunoradiometric assay in Thailand

    International Nuclear Information System (INIS)

    Kasemsuth, R.; Asavanich, A.; Sucharit, S.; Vutikes, S.; Vutikes, M.; Patamatum, S.

    1988-01-01

    The sporozoite rate, species of parasite and vector are important in the epidemiology of malaria. The investigation of sporozoite by dissection and examination under a microscope is time-consuming and it could be done only on freshly killed mosquitoes. Immunoradiometric assay (IRMA) that can detect, identify and quantify malaria sporozoite (Zavala et al., 1982) was therefore applied to detect sporozoite in laboratory-maintained Anopheles dirus and wild-caught mosquitoes. Study on P. falciparum-infected An. dirus showed that the circumsporozoite (CS) antigen was first found in the abdomen on the 10th day post-infection, whilst the sporozoites were examined in salivary glands from day 15 onwards. The malaria infection in wild-caught mosquitoes were investigated in Anopheles spp collected by human baites from three endemic areas in Thailand. Since the sporozoite rate refers to the presence of sporozoite in the salivary gland, then only head-thorax part of the specimens were detected by IRMA to prevent an exaggeration over the true results. It was found that none of mosquitoes collected from Phrae was positive for malaria. Four out of 1243 An. dirus among eight species collected from Chantaburi were positive for P. falciparum with sporozoites ranged from 270 to 3875. Of all ten species collected from Kanchanaburi, two and one out of 3123 An. minimus were positive for P. falciparum and P. vivax with sporozoites found in head-thorax portions were 1880, 2380 and 1026 respectively. It is evident that the IRMA is suitable for the investigation of malaria sporozoites in this region. The application of this technique in the further epidemiological study is in progress

  10. Validation of a quantitative Eimeria spp. PCR for fresh droppings of broiler chickens.

    Science.gov (United States)

    Peek, H W; Ter Veen, C; Dijkman, R; Landman, W J M

    2017-12-01

    A quantitative Polymerase Chain Reaction (qPCR) for the seven chicken Eimeria spp. was modified and validated for direct use on fresh droppings. The analytical specificity of the qPCR on droppings was 100%. Its analytical sensitivity (non-sporulated oocysts/g droppings) was 41 for E. acervulina, ≤2900 for E. brunetti, 710 for E. praecox, 1500 for E. necatrix, 190 for E. tenella, 640 for E. maxima, and 1100 for E. mitis. Field validation of the qPCR was done using droppings with non-sporulated oocysts from 19 broiler flocks. To reduce the number of qPCR tests five grams of each pooled sample (consisting of ten fresh droppings) per time point were blended into one mixed sample. Comparison of the oocysts per gram (OPG)-counting method with the qPCR using pooled samples (n = 1180) yielded a Pearson's correlation coefficient of 0.78 (95% CI: 0.76-0.80) and a Pearson's correlation coefficient of 0.76 (95% CI: 0.70-0.81) using mixed samples (n = 236). Comparison of the average of the OPG-counts of the five pooled samples with the mixed sample per time point (n = 236) showed a Pearson's correlation coefficient (R) of 0.94 (95% CI: 0.92-0.95) for the OPG-counting method and 0.87 (95% CI: 0.84-0.90) for the qPCR. This indicates that mixed samples are practically equivalent to the mean of five pooled samples. The good correlation between the OPG-counting method and the qPCR was further confirmed by the visual agreement between the total oocyst/g shedding patterns measured with both techniques in the 19 broiler flocks using the mixed samples.

  11. Evaluation of ionophore sensitivity of Eimeria acervulina and Eimeria maxima isolated from the Algerian Jijel province poultry farms

    Science.gov (United States)

    The present study represents the first description of ionophore resistance in Eimeria recovered from commercial Algerian (Jijel-Algeria) broiler farms. Microscopy and ITS1 PCR revealed only 2 Eimeria species present in litter from these farms- namely E. acervulina and E. maxima. A pool of these is...

  12. Dual RNA-seq reveals no plastic transcriptional response of the coccidian parasite Eimeria falciformis to host immune defenses.

    Science.gov (United States)

    Ehret, Totta; Spork, Simone; Dieterich, Christoph; Lucius, Richard; Heitlinger, Emanuel

    2017-09-05

    Parasites can either respond to differences in immune defenses that exist between individual hosts plastically or, alternatively, follow a genetically canalized ("hard wired") program of infection. Assuming that large-scale functional plasticity would be discernible in the parasite transcriptome we have performed a dual RNA-seq study of the lifecycle of Eimeria falciformis using infected mice with different immune status as models for coccidian infections. We compared parasite and host transcriptomes (dual transcriptome) between naïve and challenge infected mice, as well as between immune competent and immune deficient ones. Mice with different immune competence show transcriptional differences as well as differences in parasite reproduction (oocyst shedding). Broad gene categories represented by differently abundant host genes indicate enrichments for immune reaction and tissue repair functions. More specifically, TGF-beta, EGF, TNF and IL-1 and IL-6 are examples of functional annotations represented differently depending on host immune status. Much in contrast, parasite transcriptomes were neither different between Coccidia isolated from immune competent and immune deficient mice, nor between those harvested from naïve and challenge infected mice. Instead, parasite transcriptomes have distinct profiles early and late in infection, characterized largely by biosynthesis or motility associated functional gene groups, respectively. Extracellular sporozoite and oocyst stages showed distinct transcriptional profiles and sporozoite transcriptomes were found enriched for species specific genes and likely pathogenicity factors. We propose that the niche and host-specific parasite E. falciformis uses a genetically canalized program of infection. This program is likely fixed in an evolutionary process rather than employing phenotypic plasticity to interact with its host. This in turn might limit the potential of the parasite to adapt to new host species or niches, forcing

  13. Two new species of Eimeria Schneider, 1875 (Apicomplexa: Eimeriidae) from emerald tree skinks, Lamprolepis smaragdina (Lesson) (Sauria: Scincidae) from Papua New Guinea and the Philippines

    Science.gov (United States)

    McAllister, Chris T.; Seville, R. Scott; Duszynski, Donald W.; Bush, Sarah E.; Fisher, Robert N.; Austin, Christopher C.

    2013-01-01

    Two new species of Eimeria Schneider, 1875, from emerald tree skinks, Lamprolepis smaragdina (Lesson) are described from specimens collected in Papua New Guinea (PNG) and the Philippines. Oöcysts of Eimeria nuiailan n. sp. from the only L. smaragdina from PNG are ovoidal, with a smooth, colourless, bi-layered wall, measure 23.7 × 19.1 μm, and have a length/width (L/W) ratio of 1.3; both micropyle and oöcyst residuum are absent, but a fragmented polar granule is present. Sporocysts are ovoidal to ellipsoidal, 11.9 × 7.0 μm, L/W 1.7, and the wall is composed of two valves joined by a longitudinal suture; neither Stieda nor sub-Stieda bodies are present; a sporocyst residuum is present as a compact mass of granules. Sporozoites are elongate, 14.6 × 2.6 μm, and contain anterior and posterior refractile bodies with a nucleus between them. Oöcysts of Eimeria auffenbergi n. sp. from L. smaragdina collected in the Philippines are ovoidal, with a smooth, colourless, bi-layered wall, measure 19.9 × 15.8 μm, L/W 1.3; both micropyle and oöcyst residuum are absent, but one to four polar granules are present. Sporocysts are ovoidal to ellipsoidal, 10.3 × 5.8 μm, L/W 1.8, and the wall is composed of two valves joined by a longitudinal suture; neither Stieda nor sub-Stieda bodies are present; a sporocyst residuum is composed of dispersed granules.

  14. Two new species of Eimeria (Apicomplexa: Eimeriidae) from emerald tree skinks, Lamprolepis smaragdina (Sauria: Scincidae) from Papua New Guinea and the Philippines

    Science.gov (United States)

    McAllister, Chris T.; Seville, R. Scott; Duszynski, Donald W.; Bush, Sarah E.; Fisher, Robert N.; Austin, Christopher C.

    2014-01-01

    Two new species of Eimeria Schneider, 1875, from emerald tree skinks, Lamprolepis smaragdina (Lesson) are described from specimens collected in Papua New Guinea (PNG) and the Philippines. Oöcysts of Eimeria nuiailan sp. n. from the only L. smaragdina from PNG are ovoidal, with a smooth, colourless, bilayered wall, measure 23.7 × 19.1 μm, and have a length/width (L/W) ratio of 1.3; both micropyle and oöcyst residuum are absent, but a fragmented polar granule is present. Sporocysts are ovoidal to ellipsoidal, 11.9 × 7.0 μm, L/W 1.7, and the wall is composed of 2 valves joined by a longitudinal suture; neither Stieda nor sub-Stieda bodies are present; a sporocyst residuum is present as a compact mass of granules. Sporozoites are elongate, 14.6 × 2.6 μm, and contain anterior and posterior refractile bodies with a nucleus between them. Oöcysts of Eimeria auffenbergi sp. n. from both L. smaragdina we collected in the Philippines are ovoidal, with a smooth, colorless, bilayered wall, measure 19.9 × 15.8 μm, L/W 1.3; both micropyle and oöcyst residuum are absent, but 1–4 polar granules are present. Sporocysts are ovoidal to ellipsoidal, 10.3 × 5.8 μm, L/W 1.8, and the wall is composed of 2 valves joined by a longitudinal suture; neither Stieda nor sub-Stieda bodies are present; a sporocyst residuum is composed of dispersed granules. PMID:24048748

  15. Two new species of Eimeria Schneider, 1875 (Apicomplexa: Eimeriidae) from emerald tree skinks, Lamprolepis smaragdina (Lesson) (Sauria: Scincidae) from Papua New Guinea and the Philippines.

    Science.gov (United States)

    McAllister, Chris T; Seville, R Scott; Duszynski, Donald W; Bush, Sarah E; Fisher, Robert N; Austin, Christopher C

    2013-10-01

    Two new species of Eimeria Schneider, 1875, from emerald tree skinks, Lamprolepis smaragdina (Lesson) are described from specimens collected in Papua New Guinea (PNG) and the Philippines. Oöcysts of Eimeria nuiailan n. sp. from the only L. smaragdina from PNG are ovoidal, with a smooth, colourless, bi-layered wall, measure 23.7 × 19.1 μm, and have a length/width (L/W) ratio of 1.3; both micropyle and oöcyst residuum are absent, but a fragmented polar granule is present. Sporocysts are ovoidal to ellipsoidal, 11.9 × 7.0 μm, L/W 1.7, and the wall is composed of two valves joined by a longitudinal suture; neither Stieda nor sub-Stieda bodies are present; a sporocyst residuum is present as a compact mass of granules. Sporozoites are elongate, 14.6 × 2.6 μm, and contain anterior and posterior refractile bodies with a nucleus between them. Oöcysts of Eimeria auffenbergi n. sp. from L. smaragdina collected in the Philippines are ovoidal, with a smooth, colourless, bi-layered wall, measure 19.9 × 15.8 μm, L/W 1.3; both micropyle and oöcyst residuum are absent, but one to four polar granules are present. Sporocysts are ovoidal to ellipsoidal, 10.3 × 5.8 μm, L/W 1.8, and the wall is composed of two valves joined by a longitudinal suture; neither Stieda nor sub-Stieda bodies are present; a sporocyst residuum is composed of dispersed granules.

  16. A molecular survey of Eimeria in chickens across Australia.

    Science.gov (United States)

    Godwin, Rosamond M; Morgan, Jess A T

    2015-11-30

    Coccidiosis is a costly enteric disease of chickens caused by protozoan parasites of the genus Eimeria. Disease diagnosis and management is complicated since there are multiple Eimeria species infecting chickens and mixed species infections are common. Current control measures are only partially effective and this, combined with concerns over vaccine efficacy and increasing drug resistance, demonstrates a need for improved coccidiosis diagnosis and control. Before improvements can be made, it is important to understand the species commonly infecting poultry flocks in both backyard and commercial enterprises. The aim of this project was to conduct a survey and assessment of poultry Eimeria across Australia using genetic markers, and create a collection of isolates for each Eimeria species. A total of 260 samples (faecal or caecal) was obtained, and survey results showed that Eimeria taxa were present in 98% of commercial and 81% of backyard flocks. The distribution of each Eimeria species was widespread across Australia, with representatives of all species being found in every state and territory, and the Eimeria species predominating in commercial flocks differed from those in backyard flocks. Three operational taxonomic units also occurred frequently in commercial flocks highlighting the need to understand the impact of these uncharacterised species on poultry production. As Eimeria infections were also frequent in backyard flocks, there is a potential for backyard flocks to act as reservoirs for disease, especially as the industry moves towards free range production systems. This Eimeria collection will be an important genetic resource which is the crucial first step in the development of more sophisticated diagnostic tools and the development of new live vaccines which ultimately will provide savings to the industry in terms of more efficient coccidiosis management. Crown Copyright © 2015. Published by Elsevier B.V. All rights reserved.

  17. Memory B-Cell and Antibody Responses Induced by Plasmodium falciparum Sporozoite Immunization

    NARCIS (Netherlands)

    Nahrendorf, W.; Scholzen, A.; Bijker, E.M.; Teirlinck, A.C.; Bastiaens, G.J.H.; Schats, R.; Hermsen, C.C.; Visser, L.G.; Langhorne, J.; Sauerwein, R.W.

    2014-01-01

    BACKGROUND: Immunization of healthy volunteers during receipt of chemoprophylaxis with Plasmodium falciparum sporozoites (CPS-immunization) induces sterile protection from malaria. Antibody responses have long been known to contribute to naturally acquired immunity against malaria, but their

  18. Studies on the transfer of protective immunity with lymphoid cells from mice immune to malaria sporozoites

    International Nuclear Information System (INIS)

    Verhave, J.P.; Strickland, G.T.; Jaffe, H.A.; Ahmed, A.

    1978-01-01

    In an effort to understand the mechanisms involved in the protective immunity to malarial sporozoites, an A/J mouse/Plasmodium berghei model was studied. Protective immunity could consistently be adoptively transferred only by using sublethal irradiation of recipients (500 R); a spleen equivalent (100 x 10 6 ) of donor cells from immune syngeneic mice; and a small booster immunization (1 x 10 4 ) of recipients with irradiation-attenuated sporozoites. Recipient animals treated in this manner were protected from lethal challenge with 1 x 10 4 nonattenuated sporozoites. Immune and nonimmune serum and spleen cells from nonimmune animals did not protect recipient mice. Fewer immune spleen cells (50 x 10 6 ) protected some recipients. In vitro treatment of immune spleen cells with anti-theta sera and complement abolished their ability to transfer protection. This preliminary study suggests that protective sporozoite immunity can be transferred with cells, and that it is T cell dependent

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

    Science.gov (United States)

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

    2016-08-02

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

  20. Identification of immune protective genes of Eimeria maxima through cDNA expression library screening.

    Science.gov (United States)

    Yang, XinChao; Li, MengHui; Liu, JianHua; Ji, YiHong; Li, XiangRui; Xu, LiXin; Yan, RuoFeng; Song, XiaoKai

    2017-02-16

    Eimeria maxima is one of the most prevalent Eimeria species causing avian coccidiosis, and results in huge economic loss to the global poultry industry. Current control strategies, such as anti-coccidial medication and live vaccines have been limited because of their drawbacks. The third generation anticoccidial vaccines including the recombinant vaccines as well as DNA vaccines have been suggested as a promising alternative strategy. To date, only a few protective antigens of E. maxima have been reported. Hence, there is an urgent need to identify novel protective antigens of E. maxima for the development of neotype anticoccidial vaccines. With the aim of identifying novel protective genes of E. maxima, a cDNA expression library of E. maxima sporozoites was constructed using Gateway technology. Subsequently, the cDNA expression library was divided into 15 sub-libraries for cDNA expression library immunization (cDELI) using parasite challenged model in chickens. Protective sub-libraries were selected for the next round of screening until individual protective clones were obtained, which were further sequenced and analyzed. Adopting the Gateway technology, a high-quality entry library was constructed, containing 9.2 × 10 6 clones with an average inserted fragments length of 1.63 kb. The expression library capacity was 2.32 × 10 7 colony-forming units (cfu) with an average inserted fragments length of 1.64 Kb. The expression library was screened using parasite challenged model in chickens. The screening yielded 6 immune protective genes including four novel protective genes of EmJS-1, EmRP, EmHP-1 and EmHP-2, and two known protective genes of EmSAG and EmCKRS. EmJS-1 is the selR domain-containing protein of E. maxima whose function is unknown. EmHP-1 and EmHP-2 are the hypothetical proteins of E. maxima. EmRP and EmSAG are rhomboid-like protein and surface antigen glycoproteins of E. maxima respectively, and involved in invasion of the parasite. Our

  1. Óleo essencial de orégano, alecrim, canela e extrato de pimenta no controle de Salmonella, Eimeria e Clostridium em frangos de corte

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    Tânia D.M.M. Bona

    2012-05-01

    Full Text Available Este trabalho foi conduzido com o objetivo de avaliar a eficiência de um composto vegetal contendo óleo essencial de orégano, alecrim, canela e extrato de pimenta vermelha no controle de Salmonella, Eimeria e Clostridium em frangos de corte. Para tal, foram realizados dois experimentos. No primeiro avaliou-se a eficiência deste produto no controle de Clostridium perfringens após desafio com Eimeria acervulina, E. maxima e E. tenella. Aves de um dia de idade foram divididas em três grupos: T1 - dieta controle sem aditivo promotor de crescimento; T2 - dieta com adição de avilamicina (10ppm; e T3 - dieta com adição do composto vegetal (100ppm. O uso do composto vegetal na alimentação de frangos reduziu lesões específicas de E. maxima e E. tenella aos 14 dias pós-inoculação (PI como também reduziram a contagem de unidades formadoras de colônias (UFC de Clostridium perfringens no conteúdo do ceco das aves em relação ao grupo controle. No segundo experimento avaliou-se a eficiência deste mesmo produto em aves desafiadas com Salmonella Enteritidis. Aves de um dia de idade foram distribuídas em três tratamentos, sendo T1 - dieta controle sem adição de antibiótico promotor de crescimento, T2 - dieta com 10ppm de Avilamicina, T3 - dieta com 100ppm de um produto a base do composto vegetal acima citado. Aos 21 dias de idade todas as aves foram inoculadas com 10(5 UFC de Salmonella Enteritidis. A utilização do composto vegetal e avilamicina diminuiu a excreção de Salmonella nas aves 72 horas PI de Salmonella. A utilização do composto vegetal aumentou a relação vilo/células CD3+ no duodeno, em relação ao grupo avilamicina e controle, porém não teve efeito sobre a expressão destas células no ceco.

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

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    M. Das

    2015-08-01

    Full Text Available 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: Examination of fecal samples revealed an overall prevalence of 11.97% Eimeria infection in dairy cattle of Guwahati, Assam. Age-wise, 33.2%, 45.4%, and 21.4% infections were recorded in calves (3 years cattle, respectively. Season-wise, infection was recorded highest during post-monsoon (16.29%, followed by monsoon (15%, winter (9.44%, and pre-monsoon (7.49% season. Seven species of Eimeria were recorded viz. Eimeria bovis, Eimeria zuernii, Eimeria subspherica, Eimeria bukidnonensis, Eimeria auburnensis, Eimeria ellipsoidalis and Eimeria alabamensis. The oocyst count per gram of feces ranged from 50 to 1500 in infected cattle. Conclusion: This study indicates that there is the prevalence of seven species of Eimeria in dairy cattle of Guwahati, Assam and mostly prevalent during the post-monsoon season.

  3. Field abundance and nest structure of the ant Anoplolepis tenella associated with the African root and tuber scale in the Congo Basin

    OpenAIRE

    Fotso Kuate, A.; Tindo, M.; Hanna, R.; Kenne, M.; Goergen, G.

    2006-01-01

    Anoplolepis tenella is a ground-dwelling ant of the tropical forest zone of Central Africa. It is associated with the African root and tuber scale (ARTS), Stictococcus vayssierei Richard, an emerging cassava pest in the zone. Developing sustainable and effective methods to control ARTS requires basic knowledge of the biology of A. tenella and the nature of interactions between the two insects. A study on the nest distribution and composition of A. tenella in various vegetation types prevailin...

  4. The Plasmodium protein P113 supports efficient sporozoite to liver stage conversion in vivo.

    Science.gov (United States)

    Offeddu, Vittoria; Rauch, Manuel; Silvie, Olivier; Matuschewski, Kai

    2014-02-01

    Invasive stages of Plasmodium parasites possess distinct integral and peripheral membrane proteins that mediate host cell attachment and invasion. P113 is an abundant protein in detergent-resistant high molecular weight complexes in Plasmodium schizonts, but is unusual since expression extends to gametocytes and sporozoites. In this study, we tested whether P113 performs important functions for parasite propagation in Plasmodium berghei. We show that pre-erythrocytic expression of P113 displays key signatures of upregulated in infectious sporozoites (UIS) genes, including control by the liver stage master regulator SLARP. Targeted gene deletion resulted in viable blood stage parasites that displayed no signs of blood stage growth defects. p113(-) parasites propagated normally through the life cycle until mature sporozoites, but displayed defects during natural sporozoite transmission, leading to a delay to patency in infected animals. By comparative in vitro and in vivo analysis of pre-erythrocytic development and using a xeno-diagnostic test we show that ablation of P113 results in lower sporozoite to liver stage conversion and, as a consequence, reduced merozoite output in vivo, without delaying liver stage development. We conclude that p113 is dispensable for Plasmodium life cycle progression and plays auxiliary roles during pre-erythrocytic development. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. A sporozoite asparagine-rich protein controls initiation of Plasmodium liver stage development.

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    Olivier Silvie

    2008-06-01

    Full Text Available Plasmodium sporozoites invade host hepatocytes and develop as liver stages (LS before the onset of erythrocytic infection and malaria symptoms. LS are clinically silent, and constitute ideal targets for causal prophylactic drugs and vaccines. The molecular and cellular mechanisms underlying LS development remain poorly characterized. Here we describe a conserved Plasmodium asparagine-rich protein that is specifically expressed in sporozoites and liver stages. Gene disruption in Plasmodium berghei results in complete loss of sporozoite infectivity to rodents, due to early developmental arrest after invasion of hepatocytes. Mutant sporozoites productively invade host cells by forming a parasitophorous vacuole (PV, but subsequent remodelling of the membrane of the PV (PVM is impaired as a consequence of dramatic down-regulation of genes encoding PVM-resident proteins. These early arrested mutants confer only limited protective immunity in immunized animals. Our results demonstrate the role of an asparagine-rich protein as a key regulator of Plasmodium sporozoite gene expression and LS development, and suggest a requirement of partial LS maturation to induce optimal protective immune responses against malaria pre-erythrocytic stages. These findings have important implications for the development of genetically attenuated parasites as a vaccine approach.

  6. Long term protection after immunization with P. berghei sporozoites correlates with sustained IFNgamma responses of hepatic CD8+ memory T cells.

    NARCIS (Netherlands)

    Nganou Makamdop, C.K.; Gemert, G.J.A. van; Arens, T.; Hermsen, C.C.; Sauerwein, R.W.

    2012-01-01

    Protection against P. berghei malaria can successfully be induced in mice by immunization with both radiation attenuated sporozoites (RAS) arresting early during liver stage development, or sporozoites combined with chloroquine chemoprophylaxis (CPS), resulting in complete intra-hepatic parasite

  7. Heterologous Protection against Malaria after Immunization with Plasmodium falciparum Sporozoites.

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    Remko Schats

    Full Text Available Sterile protection in >90% of volunteers against homologous Plasmodium falciparum infection has been achieved only using the controlled human malaria infection (CHMI model. This efficient model involves whole parasite immunizations under chloroquine prophylaxis (CPS-immunization, requiring only 30-45 mosquitoes bites infected with P. falciparum-sporozoites. Given the large diversity of P. falciparum parasites, it is essential to assess protection against heterologous parasite strains.In an open-label follow-up study, 16 volunteers previously CPS-immunized and challenged with P. falciparum NF54 (West-Africa in a dose de-escalation and challenge trial were re-challenged with clone NF135.C10 (Cambodia at 14 months after the last immunization (NCT01660854.Two out of thirteen NF54 protected volunteers previously fully protected against NF54 were also fully protected against NF135.C10, while 11/13 showed a delayed patency (median prepatent period of 10.5 days (range 9.0-15.5 versus 8.5 days in 5 malaria-naïve controls (p = 0.0005. Analysis of patency by qPCR indicated a 91 to >99% estimated reduction of liver parasite load in 7/11 partially protected subjects. Three volunteers previously not protected against NF54, were also not protected against NF135.C10.This study shows that CPS-immunization can induce heterologous protection for a period of more than one year, which is a further impetus for clinical development of whole parasite vaccines.Clinicaltrials.gov NCT01660854.

  8. The complete mitochondrial genome sequence of Eimeria magna (Apicomplexa: Coccidia).

    Science.gov (United States)

    Tian, Si-Qin; Cui, Ping; Fang, Su-Fang; Liu, Guo-Hua; Wang, Chun-Ren; Zhu, Xing-Quan

    2015-01-01

    In the present study, we determined the complete mitochondrial DNA (mtDNA) sequence of Eimeria magna from rabbits for the first time, and compared its gene contents and genome organizations with that of seven Eimeria spp. from domestic chickens. The size of the complete mt genome sequence of E. magna is 6249 bp, which consists of 3 protein-coding genes (cytb, cox1 and cox3), 12 gene fragments for the large subunit (LSU) rRNA, and 7 gene fragments for the small subunit (SSU) rRNA, without transfer RNA genes, in accordance with that of Eimeria spp. from chickens. The putative direction of translation for three genes (cytb, cox1 and cox3) was the same as those of Eimeria species from domestic chickens. The content of A + T is 65.16% for E. magna mt genome (29.73% A, 35.43% T, 17.09 G and 17.75% C). The E. magna mt genome sequence provides novel mtDNA markers for studying the molecular epidemiology and population genetics of Eimeria spp. and has implications for the molecular diagnosis and control of rabbit coccidiosis.

  9. Ontogenia do fruto em desenvolvimento de Alternanthera tenella Colla e Amaranthus blitum Linnaeus (Amaranthaceae Ontogeny of fruits of Alternanthera tenella Colla and Amaranthus blitum Linnaeus (Amaranthaceae

    Directory of Open Access Journals (Sweden)

    Vanessa de Carvalho Harthman

    2012-09-01

    Full Text Available Alternanthera tenella Colla e Amaranthus blitum Linnaeus são espécies invasoras que ocorrem em culturas e terrenos baldios na região de Maringá, Paraná. O trabalho teve por objetivo a análise morfoanatômica dos frutos em desenvolvimento e estruturas não pericárpicas dessas duas espécies, com a finalidade de contribuir com informações estruturais para identificação das espécies, classificação dos frutos e investigações ecológicas. Flores e frutos foram coletados no campus da Universidade Estadual de Maringá, fixados em Glutaraldeído, secionados em micrótomo de rotação e corados com a azul de Toluidina. As bractéolas e perigônio são persistentes nos frutos e têm estrutura diferente nas duas espécies. O aquênio de Alternanthera tenella mantém o mesmo número de estratos celulares que o ovário, que sofrem colapso na fase madura, exceto o mesocarpo interno que se mantém com espessamento parietal em U e cristais. O utrículo de Amaranthus blitum é semelhante ao ovário em número de camadas celulares e apresenta aerênquima quando maduro. As sementes maduras são exotestais, com mesotesta e endotesta colapsadas, e embrião curvo. Nesse estudo, foram registrados alguns caracteres estruturais dos perigônios e dos frutos que são potencialmente significativos para caracterização e separação das espécies, ao contrário das sementes que são muito semelhantes.Alternanthera tenella Colla and Amaranthus blitum Linnaeus are weeds that occur in crops and uncultivated areas in the Maringá region of Paraná. In this study, a morphoanatomical analysis of fruit development and the pericarp of A. tenella and A. blitum was made in order to contribute structural information for species identification, fruit classification, and ecological investigations. Flowers and fruits were collected at the campus of the State University of Maringá, Paraná, fixed in glutaraldehyde, sectioned with a rotary microtome and stained with

  10. Influence of climatic and management factors on Eimeria infections in goats from semi-arid zones.

    Science.gov (United States)

    Ruiz, A; González, J F; Rodríguez, E; Martín, S; Hernández, Y I; Almeida, R; Molina, J M

    2006-10-01

    A survey of Eimeria infections was performed in dairy goats and kids (<6 months old) of six farms from a dry desert area of Gran Canaria Island (Spain). The number of oocysts per gram of faeces (OPG) was determined by a modified McMaster technique over a total of 2,616 individual faecal samples taken from the rectum in monthly intervals. Eimeria oocysts were found in 96.1% of the samples with OPG ranging from 1 x 10(2) to 1.4 x 10(6). Kid goats had significantly (P < 0.001) higher OPG counts (46,496 +/- 5,228) than dairy females (2,225 +/- 287). Eight Eimeria species were identified, with Eimeria ninakohlyakimovae (30.0%), Eimeria arloingi (28.6%) and Eimeria alijevi (20.5%) being the most frequent species followed by Eimeria caprina (9.1%), Eimeria christenseni (4.5%), Eimeria jolchijevi (3.4%), Eimeria caprovina (3.2%) and Eimeria hirci (0.7%). Although significant differences were observed among goat groups and herds, the eight species were present in the six farms in both dairy goats and kids. The intensity of oocysts shedding was related to some factors such as the size of the herd and was further influenced by the prevailing climatic conditions of the area. The highest OPG counts were recorded during the hot season in dairy goats and close to weaning time in kids reared in small farms having no prophylactic treatments against eimeriosis.

  11. Evaluation of ionophore sensitivity of Eimeria acervulina and Eimeria maxima isolated from the Algerian to Jijel province poultry farms.

    Science.gov (United States)

    Djemai, Samir; Mekroud, Abdeslam; Jenkins, Mark C

    2016-07-15

    The present study represents the first description of ionophore resistance in recovered from commercial Algerian (Jijel-Algeria) broiler farms. Microscopy and intervening transcribed sequence 1 PCR (ITS1 PCR) revealed only 2 Eimeria species present in litter from these farms- namely Eimeria acervulina and Eimeria maxima. A pool of these isolates were evaluated in broiler chickens (Cobb 500) for sensitivity to 5 anticoccidial compounds-diclazuril (1ppm), lasalocid (125ppm), monensin (125ppm), narasin (70ppm) and salinomycin (60ppm). As indicated by anticoccidial sensitivity profiles based on lesion scores and anticoccidial index (ACI), complete resistance to monensin and narasin, partial resistance to salinomycin and lasalocid, and complete sensitivity to diclazuril was observed. While lack of sensitivity to monensin is not surprising given its use for years as the sole anticoccidial compound, the resistance to monoether (narasin) and polyether (lasalocid) ionophores suggests that cross-resistance has developed in a segment of the Eimeria population. The fairly uniform Eimeria species composition among all poultry farms suggests that E. acervulina and E. maxima more rapidly develop resistance to ionophore drugs. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Co-infection of chickens with Eimeria praecox and Eimeria maxima does not prevent development of immunity to Eimeria maxima.

    Science.gov (United States)

    Jenkins, M; Fetterer, R; Miska, K

    2009-05-12

    Previous studies revealed an ameliorating effect of Eimeria praecox on concurrent E. maxima infection, such that weight gain, feed conversion ratio, and intestinal lesions were nearly identical to uninfected or E. praecox-infected controls. The purpose of the present study was to determine if protective immunity against E. maxima challenge infection developed in chickens infected with both E. praecox and E. maxima. Day-old chickens were infected with 10(3)E. praecox, 10(3)E. maxima, or a mixture of 10(3)E. praecox and 10(3)E. maxima oocysts. Chickens were then challenged at 4 weeks of age with 5x10(4)E. praecox or 5x10(3)E. maxima oocysts and clinical signs of coccidiosis were assessed 7 days post-challenge. Relative to non-challenged controls, naïve chickens or chickens immunized with E. praecox displayed a 32-34% weight gain depression after challenge with 5x10(3)E. maxima oocysts. In contrast, chickens immunized with either E. maxima oocysts alone or a combination of E. praecox and E. maxima oocysts displayed complete protection against lower weight gain associated with E. maxima challenge. Also, protection against decreased feed conversion ratio and intestinal lesions was observed in single E. maxima- or dual E. maxima+E. praecox-immunized chickens. These findings indicate that co-infection of chickens with E. maxima and E. praecox does not prevent development of immunity against E. maxima or E. praecox challenge.

  13. Identification of Eimeria acervulina conoid antigen using chicken monoclonal antibody.

    Science.gov (United States)

    Matsubayashi, Makoto; Minoura, Chisa; Kimura, Shintaro; Tani, Hiroyuki; Furuya, Masaru; Lillehoj, Hyun S; Matsuda, Haruo; Takenaka, Shigeo; Hatta, Takeshi; Tsuji, Naotoshi; Sasai, Kazumi

    2016-11-01

    In the poultry industry, Eimeria spp. is one of the important pathogens which cause significant economic losses. We have previously generated a chicken monoclonal antibody (mAb), 6D-12-G10, with specificity for an antigen located in the apical cytoskeleton of Eimeria acervulina and with cross-reactive among Apicomplexan parasites, including other Eimeria spp., Toxoplasma, Neospora, and Cryptosporidium spp. Furthermore, the protein of Cryptosporidium parvum recognized by the 6D-12-G10 has been identified as elongation factor-1α (EF-1α). In the present study, to identify the target molecule of E. acervulina by the mAb, we performed two-dimensional Western blotting analysis. Finally, we found two positive molecules which are identified as EF-1α and a related protein. Our previous finding using C. parvum and the results in this study suggest that EF-1α could be associated with the invasion facilitated by the cytoskeleton at the apical region of zoites.

  14. Environmental predictors of bovine Eimeria infection in western Kenya.

    Science.gov (United States)

    Makau, D N; Gitau, G K; Muchemi, G K; Thomas, L F; Cook, E A J; Wardrop, N A; Fèvre, E M; de Glanville, W A

    2017-02-01

    Eimeriosis is caused by a protozoan infection affecting most domestic animal species. Outbreaks in cattle are associated with various environmental factors in temperate climates but limited work has been done in tropical settings. The objective of this work was to determine the prevalence and environmental factors associated with bovine Eimeria spp. infection in a mixed farming area of western Kenya. A total of 983 cattle were sampled from 226 cattle-keeping households. Faecal samples were collected directly from the rectum via digital extraction and analysed for the presence of Eimeria spp. infection using the MacMaster technique. Individual and household level predictors of infection were explored using mixed effects logistic regression. The prevalence of individual animal Eimeria infection was 32.8% (95% CI 29.9-35.9). A positive linear relationship was found between risk of Eimeria infection and increasing temperature (OR = 1.4, 95% CI 1.06-1.86) and distance to areas at risk of flooding (OR = 1.49, 95% CI 1.17-1.91). There was weak evidence of non-linear relationship between Eimeria infection and the proportion of the area around a household that was classified as swamp (OR = 1.12, 95% CI 0.87-1.44; OR (quadratic term) = 0.85, 95% CI 0.73-1.00), and the sand content of the soil (OR = 1.18, 95% CI 0.91-1.53; OR (quadratic term) = 1.1, 95% CI 0.99-1.23). The risk of animal Eimeria spp. infection is influenced by a number of climatic and soil-associated conditions.

  15. A Study of Eimeria Species in Sheep in Mosul City

    Directory of Open Access Journals (Sweden)

    M. H. Hasan

    2012-01-01

    Full Text Available The study was conducted to diagnose and study species of Eimeria in sheep in Mosul city from beginning of September2009 to end May 2010, as well as to determine the percentage and intensity of infection of Eimeria species. Five hundredfecal samples of sheep with different ages were collected from different areas of the Mosul city. The results showed that totalpercentage of Emeria infection was 63.6%. The variations in percentage of infection were recorded according to month ofstudy. Highest percentage was recorded in March being 89.2% and the lowest in September 25.9%. The species E. ovinarecorded the highest infection rate 86.7%, while the species E. granulosa represented lowest infection rate 10%. Moreover theintensity of infection was higher in young ages and lower in adult. The results were detected that indoor sheep infection withhigh parasitic infection 69.9% whereas outdoor animals have an infection rate 25.3%. The morphological characters of oocystswere varied according to species of Eimeria has been studied. Fifty of intestinal and abomasal samples from both slaughteredin shops butchery in Mosul city and dead animals were examined to detect Eimeria infection, and results show that infectionpercentage was 56.4% in intestine of slaughtered animals and 36.3% in dead animal. Moreover no infection of Eimeria weredetected in abomasums in both slaughtered and dead animals. The oocysts of (E. parva, E.pallida and E. ovinoidalis detectedat more than 5000 oocysts per gram of intestinal contents. The intestinal secraping stained with Giemsa stain reveals thepresence of different developmental stages of parasites in wall of intestine. The histopathological sections of intestine revealedthe different pathological changes concerning of Eimeria infection.

  16. Immunolocalization of an enterotoxic glycoprotein exoantigen on the secretory organelles of Cryptosporidium parvum sporozoites

    Directory of Open Access Journals (Sweden)

    El-Shewy K.A.

    2004-06-01

    Full Text Available In this study, the fine ultrastructures of the secretory organelles of C. parvum sporozoites were demonstrated using transmission electron microscopy (TEM. Meanwhile, a previously identified enterotoxic 18-20 kDa copro-antigen (18-20 kDa CCA, associated with cryptosporidiosis in both human and calves, was isolated and immunolocalized on C. parvum sporozoites. Using immunoelectron microscopy and anti-18-20 kDa monospecific antibody demonstrated marked existence of the 18-20 kDa CCA on the apical organelles and at the trilaminar pellicles. An anterior extrusion of this protein was demonstrated around the excysted and released sporozoites. However, non excysted sporozoites did not show this protein. Affinity blotting, with biotinylated jacalin, demonstrated the O-linked oligosaccharide moiety of this protein. The potential role of this protein in the host cell invasion and/or gliding motility remains unelucidated. However, its enterotoxicity, location and secretory nature suggest that it may be a target for neutralization or invasion inhibition of Cryptosporidium.

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

    Science.gov (United States)

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

    2016-09-19

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

  18. 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...

  19. Plasmodium vivax sporozoite challenge in malaria-naïve and semi-immune Colombian volunteers

    DEFF Research Database (Denmark)

    Arévalo-Herrera, Myriam; Forero-Peña, David A.; Rubiano, Kelly

    2014-01-01

    induced in naïve and semi-immune volunteers by infected mosquito bites was compared. Methods: Seven malaria-naïve and nine semi-immune Colombian adults (n = 16) were subjected to the bites of 2-4 P. vivax sporozoite-infected Anopheles mosquitoes. Parasitemia levels, malaria clinical manifestations...

  20. A T-cell response to a liver-stage Plasmodium antigen is not boosted by repeated sporozoite immunizations

    Science.gov (United States)

    Murphy, Sean C.; Kas, Arnold; Stone, Brad C.; Bevan, Michael J.

    2013-01-01

    Development of an antimalarial subunit vaccine inducing protective cytotoxic T lymphocyte (CTL)-mediated immunity could pave the way for malaria eradication. Experimental immunization with sporozoites induces this type of protective response, but the extremely large number of proteins expressed by Plasmodium parasites has so far prohibited the identification of sufficient discrete T-cell antigens to develop subunit vaccines that produce sterile immunity. Here, using mice singly immunized with Plasmodium yoelii sporozoites and high-throughput screening, we identified a unique CTL response against the parasite ribosomal L3 protein. Unlike CTL responses to the circumsporozoite protein (CSP), the population of L3-specific CTLs was not expanded by multiple sporozoite immunizations. CSP is abundant in the sporozoite itself, whereas L3 expression does not increase until the liver stage. The response induced by a single immunization with sporozoites reduces the parasite load in the liver so greatly during subsequent immunizations that L3-specific responses are only generated during the primary exposure. Functional L3-specific CTLs can, however, be expanded by heterologous prime-boost regimens. Thus, although repeat sporozoite immunization expands responses to preformed antigens like CSP that are present in the sporozoite itself, this immunization strategy may not expand CTLs targeting parasite proteins that are synthesized later. Heterologous strategies may be needed to increase CTL responses across the entire spectrum of Plasmodium liver-stage proteins. PMID:23530242

  1. 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

  2. Genetic characterization and phylogenetic analysis of Eimeria arloingi in Iranian native kids.

    Science.gov (United States)

    Khodakaram-Tafti, A; Hashemnia, M; Razavi, S M; Sharifiyazdi, H; Nazifi, S

    2013-09-01

    Among the 16 species of Eimeria from goats, Eimeria arloingi and Eimeria ninakohlyakimovae are regarded as the most pathogenic species in the world and cause clinical caprine coccidiosis. E. arloingi is known to be an important cause of coccidiosis in Iranian kids. Molecular analyses of two portions of nuclear ribosomal DNA (internal transcribed spacer1 (ITS1) and 18S rDNA) were used for the genetic characterization of the E. arloingi. Comparison of the sequencing data of E. arloingi obtained in the present study (ITS1: KC507793 and 18S rDNA: KC507792) with other Eimeria species in the GenBank database revealed a particularly close relationship between E. arloingi and Eimeria spp. from the cattle and sheep. The phylogram based on the ITS1 sequences shows that the E. arloingi, Eimeria bovis, and Eimeria zuernii formed a distinct group separate from the other remaining Eimeria spp. in cattle and poultry. In pairwise alignment, 18S rDNA sequence derived from E. arloingi showed 99% similarity to Eimeria ahsata with differences observed at only three nucleotides. This study showed that the ITS1 and 18S rDNA gene are useful genetic markers for the specific identification and differentiation of Eimeria spp. in ruminants.

  3. Immunostimulatory complexes containing Eimeria tenella antigens and low toxicity plant saponins induce antibody response and provide protection from challenge in broiler chickens

    Science.gov (United States)

    Immunostimulating complexes (ISCOMs) are unique multimolecular structures formed by encapsulating antigens, lipids and triterpene saponins and are one of the most successful antigen delivery systems for microbial antigens. In the current study, both the route of administration and the antigen conce...

  4. Prevalence and intensity of Nematodirus sp. and Eimeria sp ...

    African Journals Online (AJOL)

    There was a significant correlation between OPG and host size. We conclude that differences in parasite loads are determined by both environmental and biological factors. KEY WORDS: goats, Nematodirus, Eimeria, season, site, food, sex, age. Egyptian Journal of Botany Vol.5 2003: 78-85. AJOL African Journals Online.

  5. New Eimeria species from the snowy owl (Nyctea scandiaca)

    Czech Academy of Sciences Publication Activity Database

    Volf, J.; Koudela, Břetislav; Modrý, D.

    1998-01-01

    Roč. 45, č. 1 (1998), s. 8 ISSN 1066-5234. [New Eimeria species from the snowy owl (Nyctea scandiaca). 01.01.1998-02.01.1998, Praha] R&D Projects: GA MŠk 8111231 Subject RIV: fp - Other Medical Disciplines

  6. Immune responses in Eimeria acervulina infected one-day-old broilers compared to amount of Eimeria in the duodenum, measured by real-time PCR

    NARCIS (Netherlands)

    Swinkels, W.J.C.; Post, J.; Cornelissen, J.B.W.J.; Engel, B.; Boerma, S.; Rebel, J.M.J.

    2006-01-01

    T-cell responses are supposed to be the major immune reactions in broilers infected with Eimeria. The nature of such T-cell responses is influenced by the species of Eimeria involved, age of the host, amount of parasites and the preceding infection history. In young chicks the intestine is still

  7. Potassium 4-(hydroxymethyl)-benzenosulfonate: a novel metabolite isolated from the marine red alga Bostrychia tenella (Rhodomelaceae, ceramiales); 4-(Hidroximetil)-Benzenossulfonato de potassio: metabolito inedito isolado da alga marinha Bostrychia tenella (Rhodomelaceae, ceramiales)

    Energy Technology Data Exchange (ETDEWEB)

    Felicio, Rafael de; Debonsi, Hosana Maria [Universidade de Sao Paulo (USP), Ribeirao Preto, SP (Brazil). Faculdade de Ciencias Farmaceuticas. Dept. de Fisica e Quimica]. E-mail: hosana@fcfrp.usp.br; Yokoya, Nair Sumie [Instituto de Botanica de Sao Paulo, SP (Brazil). Secao de Ficologia

    2008-07-01

    Chemical investigation of the dichloromethane/methanol extract of the marine alga Bostrychia tenella has led to the isolation of two aromatic compounds, the new sulfate metabolite potassium 4-(hydroxymethyl)-benzenosulfonate (1) and the compound 1-methoxyphenethyl alcohol (2), described previously as a synthetic product. Their structures were determined by spectroscopic methods including NMR, MS, IR and by comparison with literature data. (author)

  8. Transition of Plasmodium sporozoites into liver stage-like forms is regulated by the RNA binding protein Pumilio

    KAUST Repository

    Gomes-Santos, Carina S. S.; Braks, Joanna; Prudê ncio, Miguel; Carret, Cé line; Gomes, Ana Rita; Pain, Arnab; Feltwell, Theresa; Khan, Shahid; Waters, Andrew; Janse, Chris; Mair, Gunnar R.; Mota, Maria M.

    2011-01-01

    -associated environmental cues. Puf2- sporozoites exhibit genome-wide transcriptional changes that result in loss of gliding motility, cell traversal ability and reduction in infectivity, and, moreover, trigger metamorphosis typical of early Plasmodium intra-hepatic

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

    Science.gov (United States)

    Al-Badri, Riadh; Barta, John Robert

    2012-11-01

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

  10. Molecular characterization of crane Coccidia, Eimeria gruis and E. reichenowi, found in feces of migratory cranes.

    Science.gov (United States)

    Matsubayashi, Makoto; Takami, Kazutoshi; Abe, Niichiro; Kimata, Isao; Tani, Hiroyuki; Sasai, Kazumi; Baba, Eiichiroh

    2005-08-01

    Eimeria gruis and E. reichenowi have lethal pathogenicity to a number of species of cranes. These parasites develop at multiple organs or tissues in infected cranes, thus lacking the specificity of infection sites shown by other Eimeria spp. in spite of morphologic similarity. To date, there have been many reports of crane Eimeria infections, however, genetic examinations of these parasites have never been published. In the present study, we isolated oocysts of E. gruis and E. reichenowi from crane feces at a wintering area in Japan. By phylogenic analysis, we first demonstrated that partial sequences of the isolates formed their own cluster, located separately from other Eimeria spp.

  11. The complete mitochondrial genomes of five Eimeria species infecting domestic rabbits.

    Science.gov (United States)

    Liu, Guo-Hua; Tian, Si-Qin; Cui, Ping; Fang, Su-Fang; Wang, Chun-Ren; Zhu, Xing-Quan

    2015-12-01

    Rabbit coccidiosis caused by members of the genus Eimeria can cause enormous economic impact worldwide, but the genetics, epidemiology and biology of these parasites remain poorly understood. In the present study, we sequenced and annotated the complete mitochondrial (mt) genomes of five Eimeria species that commonly infect the domestic rabbits. The complete mt genomes of Eimeria intestinalis, Eimeria flavescens, Eimeria media, Eimeria vejdovskyi and Eimeria irresidua were 6261bp, 6258bp, 6168bp, 6254bp, 6259bp in length, respectively. All of the mt genomes consist of 3 genes for proteins (cytb, cox1, and cox3), 14 gene fragments for the large subunit (LSU) rRNA and 11 gene fragments for the small subunit (SSU) rRNA, but no transfer RNA (tRNA) genes. The gene order of the mt genomes is similar to that of Plasmodium, but distinct from Haemosporida and Theileria. Phylogenetic analyses based on full nucleotide sequences using Bayesian analysis revealed that the monophyly of the Eimeria of rabbits was strongly statistically supported with a Bayesian posterior probabilities. These data provide novel mtDNA markers for studying the population genetics and molecular epidemiology of the Eimeria species, and should have implications for the molecular diagnosis, prevention and control of coccidiosis in rabbits. Copyright © 2015 Elsevier Inc. All rights reserved.

  12. 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.

  13. THE SPOROZOITE ENZYME-LINKED IMMUNOSORBENT ASSAY : APPLICATION IN MALARIA EPIDEMIOLOGY

    Directory of Open Access Journals (Sweden)

    Michael J. Bangs

    2012-09-01

    Full Text Available Recent biotechnological breakthroughs have led to the development of various methods for detection and identification of human pathogens in their vectors. Monoclonal antibodies produced against malaria sporozoite antigens have permitted the development of several sensitive, species specific immunological tests (IFA, IRMA, ELIS A. One of these, a two-site enzyme-linked immunosorbent assay (ELIS A has been developed as a useful epidemiological tool in the identification of malaria-infected mosquitoes. This method employs highly species specific monoclonal antibodies that recognize the repetitive immunodominant epitope of the circumsporozoite (CS protein. Monoclonal antibodies have been developed for all four species of human malaria The key feature of the ELISA technique is the use of an enzyme indicator for an immunological reaction. The antigen capture or "sandwich" ELISA configuration uses the purified monoclonal both as the solid phase and, conjugated to enzyme, as a marker for the presence of CS protein in a mosquito homogenate incubated in the wells of a microtitration plate. This technology has shown advantages over other methods for epidemiological data collection. Mosquitoes can be caught, dried and stored until a time convenient for examination. The sporozoite rate by Plasmodium species can be identified easily, and when combined with the man-biting rate provides the sporozoite inoculation rate, an important entomologic estimate of the number of potential infective bites a person could expect over a given period of time. Presently, mosquitoes can be tested individually or pooled up to 20 anophe lines. The assay is sensitive enough to detect 1 infected mosquito per pool or as few as 25 sporozoites per 50 pi of mosquito extract. Basic principles and procedures are covered concerning solid substrate, adsorption to solid substrate, buffers and wash solutions, conjugates and enzyme substrates. The advantages and limitations of this technique

  14. Temperature shift and host cell contact up-regulate sporozoite expression of Plasmodium falciparum genes involved in hepatocyte infection.

    Directory of Open Access Journals (Sweden)

    Anthony Siau

    Full Text Available Plasmodium sporozoites are deposited in the skin by Anopheles mosquitoes. They then find their way to the liver, where they specifically invade hepatocytes in which they develop to yield merozoites infective to red blood cells. Relatively little is known of the molecular interactions during these initial obligatory phases of the infection. Recent data suggested that many of the inoculated sporozoites invade hepatocytes an hour or more after the infective bite. We hypothesised that this pre-invasive period in the mammalian host prepares sporozoites for successful hepatocyte infection. Therefore, the genes whose expression becomes modified prior to hepatocyte invasion would be those likely to code for proteins implicated in the subsequent events of invasion and development. We have used P. falciparum sporozoites and their natural host cells, primary human hepatocytes, in in vitro co-culture system as a model for the pre-invasive period. We first established that under co-culture conditions, sporozoites maintain infectivity for an hour or more, in contrast to a drastic loss in infectivity when hepatocytes were not included. Thus, a differential transcriptome of salivary gland sporozoites versus sporozoites co-cultured with hepatocytes was established using a pan-genomic P. falciparum microarray. The expression of 532 genes was found to have been up-regulated following co-culture. A fifth of these genes had no orthologues in the genomes of Plasmodium species used in rodent models of malaria. Quantitative RT-PCR analysis of a selection of 21 genes confirmed the reliability of the microarray data. Time-course analysis further indicated two patterns of up-regulation following sporozoite co-culture, one transient and the other sustained, suggesting roles in hepatocyte invasion and liver stage development, respectively. This was supported by functional studies of four hitherto uncharacterized proteins of which two were shown to be sporozoite surface

  15. A specific PfEMP1 is expressed in P. falciparum sporozoites and plays a role in hepatocyte infection

    DEFF Research Database (Denmark)

    Zanghì, Gigliola; Vembar, Shruthi S; Baumgarten, Sebastian

    2018-01-01

    Heterochromatin plays a central role in the process of immune evasion, pathogenesis, and transmission of the malaria parasite Plasmodium falciparum during blood stage infection. Here, we use ChIP sequencing to demonstrate that sporozoites from mosquito salivary glands expand heterochromatin......_SpzPfEMP1. Overall, we show that the epigenetic signature of var genes is reset in mosquito stages. Moreover, the identification of a strain-specific sporozoite PfEMP1 is highly relevant for vaccine design based on sporozoites....

  16. Expression of digestive enzymes and nutrient transporters in Eimeria acervulina-challenged layers and broilers

    Science.gov (United States)

    Avian coccidiosis is a disease caused by the intestinal protozoa Eimeria. Eimeria-infected chickens develop lesions in the intestinal mucosa, which result in reduced feed efficiency and body weight gain. This growth reduction may be due to changes in expression of digestive enzymes and nutrient tran...

  17. Eimeria spp. from Japanese quails (Coturnix japonica: new characteristic features and diagnostic tools

    Directory of Open Access Journals (Sweden)

    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. The Puf-family RNA-binding protein Puf2 controls sporozoite conversion to liver stages in the malaria parasite.

    Directory of Open Access Journals (Sweden)

    Katja Müller

    Full Text Available Malaria is a vector-borne infectious disease caused by unicellular, obligate intracellular parasites of the genus Plasmodium. During host switch the malaria parasite employs specialized latent stages that colonize the new host environment. Previous work has established that gametocytes, sexually differentiated stages that are taken up by the mosquito vector, control expression of genes required for mosquito colonization by translational repression. Sexual parasite development is controlled by a DEAD-box RNA helicase of the DDX6 family, termed DOZI. Latency of sporozoites, the transmission stage injected during an infectious blood meal, is controlled by the eIF2alpha kinase IK2, a general inhibitor of protein synthesis. Whether RNA-binding proteins participate in translational regulation in sporozoites remains to be studied. Here, we investigated the roles of two RNA-binding proteins of the Puf-family, Plasmodium Puf1 and Puf2, during sporozoite stage conversion. Our data reveal that, in the rodent malaria parasite P. berghei, Puf2 participates in the regulation of IK2 and inhibits premature sporozoite transformation. Inside mosquito salivary glands puf2⁻ sporozoites transform over time to round forms resembling early intra-hepatic stages. As a result, mutant parasites display strong defects in initiating a malaria infection. In contrast, Puf1 is dispensable in vivo throughout the entire Plasmodium life cycle. Our findings support the notion of a central role for Puf2 in parasite latency during switch between the insect and mammalian hosts.

  19. Transition of Plasmodium sporozoites into liver stage-like forms is regulated by the RNA binding protein Pumilio

    KAUST Repository

    Gomes-Santos, Carina S. S.

    2011-05-19

    Many eukaryotic developmental and cell fate decisions that are effected post-transcriptionally involve RNA binding proteins as regulators of translation of key mRNAs. In malaria parasites (Plasmodium spp.), the development of round, non-motile and replicating exo-erythrocytic liver stage forms from slender, motile and cell-cycle arrested sporozoites is believed to depend on environmental changes experienced during the transmission of the parasite from the mosquito vector to the vertebrate host. Here we identify a Plasmodium member of the RNA binding protein family PUF as a key regulator of this transformation. In the absence of Pumilio-2 (Puf2) sporozoites initiate EEF development inside mosquito salivary glands independently of the normal transmission-associated environmental cues. Puf2- sporozoites exhibit genome-wide transcriptional changes that result in loss of gliding motility, cell traversal ability and reduction in infectivity, and, moreover, trigger metamorphosis typical of early Plasmodium intra-hepatic development. These data demonstrate that Puf2 is a key player in regulating sporozoite developmental control, and imply that transformation of salivary gland-resident sporozoites into liver stage-like parasites is regulated by a post-transcriptional mechanism. 2011 Gomes-Santos et al.

  20. First amplification of Eimeria hessei DNA from the lesser horseshoe bat (Rhinolophus hipposideros) and its phylogenetic relationships with Eimeria species from other bats and rodents.

    Science.gov (United States)

    Afonso, Eve; Baurand, Pierre-Emmanuel; Tournant, Pierline; Capelli, Nicolas

    2014-04-01

    Although coccidian parasites of the genus Eimeria are among the best-documented parasites in bats, few Eimeria species found in bats have been characterised using molecular tools, and none of the characterised species are found in European countries. Phylogenetic relationships of Eimeria species that parasitise bats and rodents can be related to the morphology of oocysts, independently from host range, suggesting that these species are derived from common ancestors. In the present study, we isolated a partial sequence of the Eimeria hessei 18S rRNA gene from the lesser horseshoe bat (Rhinolophus hipposideros), a European bat species. Droppings from lesser horseshoe bats were collected from 11 maternity roosts located in France that were positive for the presence of the parasite. Through morphological characterisation, the oocysts detected in the lesser horseshoe bat droppings were confirmed to be E. hessei. The unique E. hessei sequence obtained through molecular analysis belonged to a clade that includes both rodent and bat Eimeria species. However, the E. hessei oocysts isolated from the bat droppings did not show morphological similarities to rodent Eimeria species. Copyright © 2014 Elsevier Inc. All rights reserved.

  1. Intestinal mucosa structure of broiler chickens infected experimentally with Eimeria tenella and treated with essential oil of oregano Morfometria intestinal de frangos de corte infectados experimentalmente com Eimeria tenella e tratados com óleo essencial de orégano

    Directory of Open Access Journals (Sweden)

    Maria Aparecida da Silva

    2009-08-01

    Full Text Available In the first trial a total of 250 day-old male chicks were distributed into five treatments and given the following diets: a diet with growth promoter; a diet without added growth promoter; a diet added with avilamycin only; diet supplemented with 0.5g of oregano oil kg diet-1; 1.0g of oregano oil kg diet-1. In other trial a total of 288 day-old chicks was used and distributed into four treatments, which were given the following diets: a diet with anticoccidial agent; a diet without anticoccidial agent; a diet supplemented with 0.5g of oregano oil kg diet-1; a 1.0g of oregano oil kg diet-1. In the first trial the nonmedicated group had the highest crypt depth which differs from chickens fed with growth promoter or with 0.5 and 1.0g of oregano oil kg diet-1. The broilers fed with positive control (antibiotic and anticoccidial had the highest villous: crypt ratio compared with the negative control that had the lowest villous:crypt ratio and the highest oocyst excretion in litter (PInicialmente, foram utilizados, neste estudo, 250 pintos de um dia de idade distribuídos em cinco tratamentos: dieta com promotor de crescimento; dieta sem promotor de crescimento; dieta contendo somente antibiótico; dieta com 0,5g de orégano óleo kg de ração-1 ou com 1,0g de orégano óleo kg de ração-1. No outro ensaio, foram utilizados 288 pintos de um dia de idade distribuídos em quatro grupos: dieta com anticoccidiano; dieta sem anticoccidiano; dieta com 0,5g de orégano óleo kg dieta-1 ou 1,0g de orégano óleo kg de ração-1. No primeiro ensaio, o grupo tratado sem promotor de crescimento apresentou a maior profundidade de cripta quando comparada com os animais tratados com promotor de crescimento ou com 0,5 e 1,0g de orégano óleo kg de ração-1. Os frangos que receberam a dieta com promotor de crescimento (antibiótico+anticoccidiano apresentaram uma maior relação vilo:cripta em comparação com os frangos do controle negativo, os quais tiveram a menor relação vilo:cripta e uma maior excreção de oocistos por grama de fezes (P<0.05. No segundo ensaio, observou-se que os frangos alimentados com dieta sem anticoccidiano tiveram uma maior espessura de lâmina própria cecal, diferindo dos frangos tratados com anticoccidiano ou com 1,0 de orégano óleo kg de ração-1 (P <0,05.

  2. Antibacterial, antifungal and cytotoxic activities exhibited by endophytic fungi from the Brazilian marine red alga Bostrychia tenella (Ceramiales

    Directory of Open Access Journals (Sweden)

    Rafael de Felício

    Full Text Available Abstract Marine environment is one of the most important sources regarding natural products research. Besides, marine microorganisms have been denominated as a talented natural source for discovery of new leads. Although the association of macroalgae and fungi has been described regarding ecological issues, there is a lack of studies about marine seaweed endophytic fungi. In this context, the goal of this study was to evaluate cytotoxic, antifungal and antibacterial activities of endophytic fungi isolated from the Brazilian marine seaweed Bostrychia tenella (J.V. Lamouroux J. Agardh (Ceramiales, Rhodophyta. Forty-five endophytic microorganism strains were isolated from B. tenella. Crude extracts and organic fractions of ten selected strains were obtained after growth in rice medium. Samples were evaluated for cytotoxicity, antifungal and antibacterial assays. Penicillium strains showed positive results in a diversity of assays, and other five strains were active in at least one test. In addition, cytochalasin D was isolated from Xylaria sp. This alga is composed of a microbiological potential, since its endophytic strains exhibited remarkable biological properties. Moreover, cytochalasin D isolation has confirmed chemical potential of marine endophytic strains. This is the first study in which cultured fungi isolates from the Brazilian macroalga B. tenella were evaluated concerning biological properties. Results corroborated that this species could be a pharmaceutical source from marine environment. Furthermore, Acremonium implicatum is being firstly described as marine endophyte and Xylaria sp., Trichoderma atroviride and Nigrospora oryzae as marine seaweed endophytes. Thus, this work reports the first study relating detailed isolation, cultivation and biological evaluation (cytotoxic, antifungal and antibacterial of endophytes Penicillium decaturense and P. waksmanii from the Brazilian marine red alga B. tenella. We are also reporting the

  3. Evaluation of immunomodulatory and anti-inflammatory effects and phytochemical screening of Alternanthera tenella Colla (Amaranthaceae aqueous extracts

    Directory of Open Access Journals (Sweden)

    Carla de Agostino Biella

    2008-09-01

    Full Text Available Alternanthera tenella Colla extracts are used in Brazilian traditional folk medicine to treat a variety of infectious diseases as well as inflammation and fever. In this work, the immunomodulatory, anti-inflammatory and potential toxic effects of cold (CAE and hot (HAE aqueous extracts of A. tenella were investigated in vivo. In addition, we analyzed the phytochemical properties of both extracts. BALB/c mice were immunized in vivo with sheep red blood cells and concomitantly inoculated intraperitoneally (i.p. with each extract (50, 100 or 200 mg/kg. Specific antibody-producing cells were enumerated using plaque-forming cell assays (PFC and anti-SRBC IgG and IgM serum levels were measured via enzyme-linked immunosorbent assay. Body and lymphoid organ weights were determined after treatments in order to evaluate toxic effects. Carrageenan-induced paw edema was employed to investigate anti-inflammatory activity in mice inoculated i.p. with CAE or HAE (200 or 400 mg/kg. Phytochemical screening was performed using spectrometric and chromatographic approaches and revealed that CAE possessed higher tannin and flavonoid levels than HAE. PFC numbers were increased after treatment with CAE (100 mg/kg four days after immunization, as were the serum antibody titers after four and seven days, suggesting immunostimulatory activity through modulation of B lymphocyte functions. Body and organ weights did not show major changes, suggesting that extracts administered to mice did not induce significant toxicity. Both extracts had significant anti-inflammatory activity in the paw edema assay. These results suggested that aqueous extracts from A. tenella contained several chemical compounds that possess positive and/or negative modulator effects on the immune system, which appeared to correlate with tannin and flavonoid levels in those extracts. In summary, these studies provide important insight into the biological activities of A. tenella.

  4. Prevalence and molecular characterisation of Eimeria species in Ethiopian village chickens.

    Science.gov (United States)

    Luu, Lisa; Bettridge, Judy; Christley, Robert M; Melese, Kasech; Blake, Damer; Dessie, Tadelle; Wigley, Paul; Desta, Takele T; Hanotte, Olivier; Kaiser, Pete; Terfa, Zelalem G; Collins, Marisol; Lynch, Stacey E

    2013-10-15

    Coccidiosis, caused by species of the apicomplexan parasite Eimeria, is a major disease of chickens. Eimeria species are present world-wide, and are ubiquitous under intensive farming methods. However, prevalence of Eimeria species is not uniform across production systems. In developing countries such as Ethiopia, a high proportion of chicken production occurs on rural smallholdings (i.e. 'village chicken production') where infectious diseases constrain productivity and surveillance is low. Coccidiosis is reported to be prevalent in these areas. However, a reliance on oocyst morphology to determine the infecting species may impede accurate diagnosis. Here, we used cross-sectional and longitudinal studies to investigate the prevalence of Eimeria oocyst shedding at two rural sites in the Ethiopian highlands. Faecal samples were collected from 767 randomly selected chickens in May or October 2011. In addition, 110 chickens were sampled in both May and October. Eimeria oocysts were detected microscopically in 427 (56%, 95% confidence interval (95% CI) 52-59%) of the 767 faecal samples tested. Moderate clustering of positive birds was detected within households, perhaps suggesting common risk factors or exposure pathways. Seven species of Eimeria were detected by real time PCR in a subset of samples further analysed, with the prevalence of some species varying by region. Co-infections were common; 64% (23/36, 95% CI 46-79%) of positive samples contained more than one Eimeria spp. Despite frequent infection and co-infection overt clinical disease was not reported. Eimeria oocysts were detected significantly more frequently in October (248/384, 65%, 95% CI 60-69%), following the main rainy season, compared to May (179/383, 47%, 95% CI 42-52%, p Eimeria oocyst positivity in May did not significantly affect the likelihood of detecting Eimeria oocyst five months later perhaps suggesting infection with different species or immunologically distinct strains. Eimeria spp oocysts

  5. Development of a metabolically active, non-replicating sporozoite vaccine to prevent Plasmodium falciparum malaria.

    Science.gov (United States)

    Hoffman, Stephen L; Billingsley, Peter F; James, Eric; Richman, Adam; Loyevsky, Mark; Li, Tao; Chakravarty, Sumana; Gunasekera, Anusha; Chattopadhyay, Rana; Li, Minglin; Stafford, Richard; Ahumada, Adriana; Epstein, Judith E; Sedegah, Martha; Reyes, Sharina; Richie, Thomas L; Lyke, Kirsten E; Edelman, Robert; Laurens, Matthew B; Plowe, Christopher V; Sim, B Kim Lee

    2010-01-01

    Immunization of volunteers by the bite of mosquitoes carrying radiation-attenuated Plasmodium falciparum sporozoites protects greater than 90% of such volunteers against malaria, if adequate numbers of immunizing biting sessions and sporozoite-infected mosquitoes are used. Nonetheless, until recently it was considered impossible to develop, license and commercialize a live, whole parasite P. falciparum sporozoite (PfSPZ) vaccine. In 2003 Sanaria scientists reappraised the potential impact of a metabolically active, non-replicating PfSPZ vaccine, and outlined the challenges to producing such a vaccine. Six years later, significant progress has been made in overcoming these challenges. This progress has enabled the manufacture and release of multiple clinical lots of a 1(st) generation metabolically active, non-replicating PfSPZ vaccine, the Sanaria PfSPZ Vaccine, submission of a successful Investigational New Drug application to the US Food and Drug Administration, and initiation of safety, immunogenicity and protective efficacy studies in volunteers in MD, US. Efforts are now focused on how best to achieve submission of a successful Biologics License Application and introduce the vaccine to the primary target population of African children in the shortest possible period of time. This will require implementation of a systematic, efficient clinical development plan. Short term challenges include optimizing the (1) efficiency and scale up of the manufacturing process and quality control assays, (2) dosage regimen and method of administration, (3) potency of the vaccine, and (4) logistics of delivering the vaccine to those who need it most, and finalizing the methods for vaccine stabilization and attenuation. A medium term goal is to design and build a facility for manufacturing highly potent and stable vaccine for pivotal Phase 3 studies and commercial launch.

  6. Fruit and seed biometry of cambuí (Myrciaria tenella O. Berg = Biometria de fruto e semente de cambuí (Myrciaria tenella O. Berg

    Directory of Open Access Journals (Sweden)

    Ana Veruska Cruz da Silva

    2012-12-01

    Full Text Available The cambuí tree (Myrciaria tenella O. Berg is native to Brazil and its fruits are harvested by extraction for fresh consumption in the state of Sergipe (Brazil. Because of the regional importance of the species, this work aims to characterize the cambuí fruits and seeds. The fruits were harvested mature from native trees in the Reserva do Caju Experimental Field, on Itaporanga d’Ajuda (Sergipe, Brazil, belonging to Embrapa Tabuleiros Costeiros. Two hundred fruits and seeds were biometrically analyzed. In addition, we evaluated the fruit, pulp, and seed color based on RHS Color Chart. Two different skin colors were found: 1. yellow [orange - red (group 32A] with yellow pulp [yellow - orange (group 17A]; and 2. purple [violet -blue (group 93A] with red pulp [red - purple (group 60A]. The seeds have only one color, green [yellow-green (group 152A] with dark stripes [Brown (group 200C]. The fruits mean diameter was 9.23 mm.fruit-1, and average width 8.50 mm.fruit-1. The seeds mean diameter was 5.34 mm.seed-1, mean width 6.52 mm.seed-1 and thickness 5.08 mm.seed-1. The cambuí is a little red fruit. The biometry had a high variability as expected, because it is a native species without any type of management.=Resumo - O cambuí (Myrciaria tenella O. Berg é nativo do Brasil e os frutos são colhidos por extrativismo para consumo in natura no estado de Sergipe (Brasil. Por causa da importância regional da espécie, objetivou-se com este trabalho caracterizar os frutos e sementes de cambuí. Os frutos foram colhidos maduros provenientes de plantas nativas do Campo Experimental Reserva do Caju, no município de Itaporanga d’Ajuda (Sergipe, Brasil pertencente a Embrapa Tabuleiros Costeiros. Foram analisados 200 frutos e sementes biometricamente. Além disso, caracterizou-se a cor da fruta, da polpa, e da semente com base na cartela de cores RHS. Foram identificados duas cores diferentes para a fruta: 1. de cor amarela [laranja - vermelho (grupo32A

  7. Immunity to sporozoite-induced malaria infection in mice. I. The effect of immunization of T and B cell-deficient mice

    International Nuclear Information System (INIS)

    Chen, D.H.; Tigelaar, R.E.; Weinbaum, F.I.

    1977-01-01

    The cellular basis of immunity to sporozoites was investigated by examining the effect of immunization of T and B cell-deficient C57BL/6N x BALB/c AnN F 1 (BLCF 1 ) mice compared to immunocompetent controls. Immunization of T cell-deficient (ATX-BM-ATS) BLCF 1 mice with x-irradiated sporozoites did not result in the generation of protective immunity. The same immunization protocols protected all immunocompetent controls. In contrast, B cell-deficient (μ-suppressed) BLCF 1 mice were protected by immunization in the majority of cases. The absence of detectable serum circumsporozoite precipitins or sporozoite neutralizing activity in the μ-suppressed mice that resisted a sporozoite challenge suggests a minor role for these humoral factors in protection. These data demonstrate a preeminent role for T cells in the induction of protective immunity in BLCF 1 mice against a P. berghei sporozoite infection

  8. An optimised protocol for molecular identification of Eimeria from chickens☆

    Science.gov (United States)

    Kumar, Saroj; Garg, Rajat; Moftah, Abdalgader; Clark, Emily L.; Macdonald, Sarah E.; Chaudhry, Abdul S.; Sparagano, Olivier; Banerjee, Partha S.; Kundu, Krishnendu; Tomley, Fiona M.; Blake, Damer P.

    2014-01-01

    Molecular approaches supporting identification of Eimeria parasites infecting chickens have been available for more than 20 years, although they have largely failed to replace traditional measures such as microscopy and pathology. Limitations of microscopy-led diagnostics, including a requirement for specialist parasitological expertise and low sample throughput, are yet to be outweighed by the difficulties associated with accessing genomic DNA from environmental Eimeria samples. A key step towards the use of Eimeria species-specific PCR as a sensitive and reproducible discriminatory tool for use in the field is the production of a standardised protocol that includes sample collection and DNA template preparation, as well as primer selection from the numerous PCR assays now published. Such a protocol will facilitate development of valuable epidemiological datasets which may be easily compared between studies and laboratories. The outcome of an optimisation process undertaken in laboratories in India and the UK is described here, identifying four steps. First, samples were collected into a 2% (w/v) potassium dichromate solution. Second, oocysts were enriched by flotation in saturated saline. Third, genomic DNA was extracted using a QIAamp DNA Stool mini kit protocol including a mechanical homogenisation step. Finally, nested PCR was carried out using previously published primers targeting the internal transcribed spacer region 1 (ITS-1). Alternative methods tested included sample processing in the presence of faecal material, DNA extraction using a traditional phenol/chloroform protocol, the use of SCAR multiplex PCR (one tube and two tube versions) and speciation using the morphometric tool COCCIMORPH for the first time with field samples. PMID:24138724

  9. 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

  10. Self-incompatibility of Prunus tenella and evidence that reproductively isolated species of Prunus have different SFB alleles coupled with an identical S-RNase allele.

    Science.gov (United States)

    Surbanovski, Nada; Tobutt, Kenneth R; Konstantinović, Miroslav; Maksimović, Vesna; Sargent, Daniel J; Stevanović, Vladimir; Bosković, Radovan I

    2007-05-01

    Many species of Prunus display an S-RNase-based gametophytic self-incompatibility (SI), controlled by a single highly polymorphic multigene complex termed the S-locus. This comprises tightly linked stylar- and pollen-expressed genes that determine the specificity of the SI response. We investigated SI of Prunus tenella, a wild species found in small, isolated populations on the Balkan peninsula, initially by pollination experiments and identifying stylar-expressed RNase alleles. Nine P. tenella S-RNase alleles (S(1)-S(9)) were cloned; their sequence analysis showed a very high ratio of non-synonymous to synonymous nucleotide substitutions (K(a)/K(s)) and revealed that S-RNase alleles of P. tenella, unlike those of Prunus dulcis, show positive selection in all regions except the conserved regions and that between C2 and RHV. Remarkably, S(8)-RNase, was found to be identical to S(1)-RNase from Prunus avium, a species that does not interbreed with P. tenella and, except for just one amino acid, to S(11) of P. dulcis. However, the corresponding introns and S-RNase-SFB intergenic regions showed considerable differences. Moreover, protein sequences of the pollen-expressed SFB alleles were not identical, harbouring 12 amino-acid replacements between those of P. tenella SFB(8) and P. avium SFB(1). Implications of this finding for hypotheses about the evolution of new S-specificities are discussed.

  11. Baculovirus virions displaying Plasmodium berghei circumsporozoite protein protect mice against malaria sporozoite infection

    International Nuclear Information System (INIS)

    Yoshida, Shigeto; Kondoh, Daisuke; Arai, Eriko; Matsuoka, Hiroyuki; Seki, Chisato; Tanaka, Takao; Okada, Masaji; Ishii, Akira

    2003-01-01

    The display of foreign proteins on the surface of baculovirus virions has provided a tool for the analysis of protein-protein interactions and for cell-specific targeting in gene transfer applications. To evaluate the baculovirus display system as a vaccine vehicle, we have generated a recombinant baculovirus (AcNPV-CSPsurf) that displays rodent malaria Plasmodium berghei circumsporozoite protein (PbCSP) on the virion surface as a fusion protein with the major baculovirus envelope glycoprotein gp64. The PbCSP-gp64 fusion protein was incorporated and oligomerized on the virion surface and led to a 12-fold increase in the binding activity of AcNPV-CSPsurf virions to HepG2 cells. Immunization with adjuvant-free AcNPV-CSPsurf virions induced high levels of antibodies and gamma interferon-secreting cells against PbCSP and protected 60% of mice against sporozoite challenge. These data demonstrate that AcNPV-CSPsurf displays sporozoite-like PbCSP on the virion surface and possesses dual potentials as a malaria vaccine candidate and a liver-directed gene delivery vehicle

  12. Distinct Prominent Roles for Enzymes of Plasmodium berghei Heme Biosynthesis in Sporozoite and Liver Stage Maturation

    Science.gov (United States)

    Matuschewski, Kai; Haussig, Joana M.

    2016-01-01

    Malarial parasites have evolved complex regulation of heme supply and disposal to adjust to heme-rich and -deprived host environments. In addition to its own pathway for heme biosynthesis, Plasmodium likely harbors mechanisms for heme scavenging from host erythrocytes. Elaborate compartmentalization of de novo heme synthesis into three subcellular locations, including the vestigial plastid organelle, indicates critical roles in life cycle progression. In this study, we systematically profile the essentiality of heme biosynthesis by targeted gene deletion of enzymes in early steps of this pathway. We show that disruption of endogenous heme biosynthesis leads to a first detectable defect in oocyst maturation and sporogony in the Anopheles vector, whereas blood stage propagation, colonization of mosquito midguts, or initiation of oocyst development occurs indistinguishably from that of wild-type parasites. Although sporozoites are produced by parasites lacking an intact pathway for heme biosynthesis, they are absent from mosquito salivary glands, indicative of a vital role for heme biosynthesis only in sporozoite maturation. Rescue of the first defect in sporogony permitted analysis of potential roles in liver stages. We show that liver stage parasites benefit from but do not strictly depend upon their own aminolevulinic acid synthase and that they can scavenge aminolevulinic acid from the host environment. Together, our experimental genetics analysis of Plasmodium enzymes for heme biosynthesis exemplifies remarkable shifts between the use of endogenous and host resources during life cycle progression. PMID:27600503

  13. Anticoccidial activities of Chitosan on Eimeria papillata-infected mice.

    Science.gov (United States)

    Abdel-Latif, Mahmoud; Abdel-Haleem, Heba M; Abdel-Baki, Abdel-Azeem S

    2016-07-01

    Eimeria spp. multiply within the intestinal tract causing severe inflammatory responses. Chitosan (CS), meanwhile, has been shown to exhibit anti-inflammatory activities in different experimental models. Here, we investigated the effect of CS on the outcome of inflammation caused by Eimeria papillata in the mouse intestine. Investigations were undertaken into the oocyst output in feces and developmental stages and goblet cells in intestinal tissue. Assays for lipid peroxidation, nitric oxide (NO), and myeloperoxidase (MPO) were also performed. T cells in intestinal tissue were counted using immunohistochemistry while total IgA in serum or intestinal wash was assayed using ELISA. In addition, mRNA expression of tumor necrosis factor alpha (TNF-α), transforming growth factor β (TGF-β), interleukin (IL)-10, and IL-4 were detected using real-time PCR. The data indicated a reduction in both oocyst output and in the number of parasite developmental stages following CS treatment, while the goblet cell hypoplasia in infected mice was also inhibited. CS decreased lipid peroxidation, NO, and MPO but did not alter the T cell count or IgA levels in comparison to the infected group. The expression of TNF-α and TGF-β decreased but IL-10 and IL-4 increased after CS treatment in comparison to the non-treated infected group. In conclusion, CS showed anti-inflammatory and protective effects against E. papillata infection.

  14. Assessment and optimization of theileria parva sporozoite full-length p67 antigen expression in mammalian cells

    Science.gov (United States)

    Delivery of various forms of recombinant Theileria parva sporozoite antigen (p67) has been shown to elicit antibody responses in cattle capable of providing protection against East Coast fever, the clinical disease caused by T. parva. Previous formulations of full-length and shorter recombinant vers...

  15. Expression of human CD81 differently affects host cell susceptibility to malaria sporozoites depending on the Plasmodium species.

    NARCIS (Netherlands)

    Silvie, O.; Greco, C.; Franetich, J.F.; Dubart-Kupperschmitt, A.; Hannoun, L.; Gemert, G.J.A. van; Sauerwein, R.W.; Levy, S.; Boucheix, C.; Rubinstein, E.; Mazier, D.

    2006-01-01

    Plasmodium sporozoites can enter host cells by two distinct pathways, either through disruption of the plasma membrane followed by parasite transmigration through cells, or by formation of a parasitophorous vacuole (PV) where the parasite further differentiates into a replicative exo-erythrocytic

  16. Genome-wide association study and biological pathway analysis for response to Eimeria maxima in broilers

    DEFF Research Database (Denmark)

    Hamzic, Edin; Buitenhuis, Albert Johannes; Hérault, Frédéric

    2015-01-01

    Background Coccidiosis is the most common and costly disease in the poultry industry and which caused by protozoans from the genus of Eimeria. The current control of coccidiosis, based on the use of anticoccidial drugs and vaccination, faces serious obstacles such as drug resistance and the high...... costs for development of efficient vaccines, respectively. Therefore, the present control programs must be expanded with complementary approaches such as the use of genetics for improvement of the host’s response to Eimeria infections. Recently, we have performed a large-scale challenge study on Cobb500...... of the measured traits in the response to Eimeria maxima in broilers. Furthermore, we conducted a post-GWAS functional analysis with the aim of gaining a better biological understanding of the underlying response to Eimeria maxima challenge in broilers. Results In total, we identified 22 single nucleotide...

  17. ABEJAS VISITANTES DE Aspilia tenella (KUNTH S. F. BLAKE (ASTERACEAE: COMPORTAMIENTO DE FORRAJEO Y CARGAS POLÍNICAS BEES VISITING Aspilia tenella (KUNTH S .F. BLAKE (ASTERACEAE: FORAGING BEHAVIOR AND POLLEN LOADS

    Directory of Open Access Journals (Sweden)

    Clara Isabel Aguilar Sierra

    2008-12-01

    Full Text Available En cuatro localidades de la zona de influencia del embalse Porce II (Antioquia, Colombia, se observó el comportamiento de 23 taxones de abejas durante sus visitas a Aspilia tenella (Kunth S. F. Blake; se registró el número de individuos y el tiempo total de visita y se midieron la temperatura y la humedad relativa. Los análisis de varianza indican que hubo una relación estadísticamente significativa en cuanto a los atributos considerados para las especies de abejas y una covariación significativa con las variables ambientales. Las pruebas de rangos múltiples muestran que Trigona nigerrima es la especie con los mayores valores promedio para el número de individuos y el tiempo de visita. En el estudio de las cargas polínicas se encontraron 30 tipos polínicos, dentro de los cuales A. tenella presentó el mayor porcentaje; Piper aduncum, Miconia minuti flora, Psidium guajava, Rapanea sp., Mimosa pudica y Psychotria sp., en su orden, son otras fuentes de polen importantes para varias de las especies de abejas. El análisis de agrupamiento para las abejas capturadas permitió diferenciar dos grupos: el primero incluye a 18 especies cuyas cargas polínicas que poseían una abundancia relativa de granos de polen de A. tenella por encima de 77%; el segundo grupo estaba conformado por cinco especies de abejas que recolectaron porcentajes mayores de otros tipos polínicos: Exomalopsis sp. 1 (90,4% de M. minuti flora; Lasioglossum sp. 1 y Coelioxys aff. mexicana (77,4% y 64,1% de P. aduncum, respectivamente; Exomalopsis sp. 2 (67,1% de P. guajava y Auglochloropsis vesta (55,5% de Rapanea sp.. Trigona fulviventris y Apis mellifera presentaron los valores más altos de riqueza de tipos polínicos en las cargas de polen; ello evidencia sus nichos tróficos amplios y su comportamiento generalista de visita.We observed the behavior of 23 species of wild bees visiting Aspilia tenella (Kunth S .F. Blake in four different localities of the Porce II dam

  18. Evaluation of next generation sequencing for the analysis of Eimeria communities in wildlife.

    Science.gov (United States)

    Vermeulen, Elke T; Lott, Matthew J; Eldridge, Mark D B; Power, Michelle L

    2016-05-01

    Next-generation sequencing (NGS) techniques are well-established for studying bacterial communities but not yet for microbial eukaryotes. Parasite communities remain poorly studied, due in part to the lack of reliable and accessible molecular methods to analyse eukaryotic communities. We aimed to develop and evaluate a methodology to analyse communities of the protozoan parasite Eimeria from populations of the Australian marsupial Petrogale penicillata (brush-tailed rock-wallaby) using NGS. An oocyst purification method for small sample sizes and polymerase chain reaction (PCR) protocol for the 18S rRNA locus targeting Eimeria was developed and optimised prior to sequencing on the Illumina MiSeq platform. A data analysis approach was developed by modifying methods from bacterial metagenomics and utilising existing Eimeria sequences in GenBank. Operational taxonomic unit (OTU) assignment at a high similarity threshold (97%) was more accurate at assigning Eimeria contigs into Eimeria OTUs but at a lower threshold (95%) there was greater resolution between OTU consensus sequences. The assessment of two amplification PCR methods prior to Illumina MiSeq, single and nested PCR, determined that single PCR was more sensitive to Eimeria as more Eimeria OTUs were detected in single amplicons. We have developed a simple and cost-effective approach to a data analysis pipeline for community analysis of eukaryotic organisms using Eimeria communities as a model. The pipeline provides a basis for evaluation using other eukaryotic organisms and potential for diverse community analysis studies. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Eimeria spp. infecting quenda (Isoodon obesulus) in the greater Perth region, Western Australia.

    Science.gov (United States)

    Hillman, Alison E; Yang, Rongchang; Lymbery, Alan J; Thompson, R C Andrew

    2016-11-01

    Parasites of wildlife inhabiting urbanised and peri-urban environments are of interest regarding wildlife population health, and also veterinary public health in the case of parasites that can also infect humans and domestic animals. This study aimed to: identify, and estimate the prevalence of, species of Eimeria parasitic in quenda (Isoodon obesulus) in the greater Perth region, Western Australia; 2) morphologically describe and genetically characterise a novel observed species of Eimeria as E. angustus; and 3) genetically characterise E. kanyana. Eimeria spp. prevalence was 76.1% (95% CI 64.9-84.5%), and four putative species of Eimeria were identified. Eimeria kanyana was identified infecting quenda for the first time, with a prevalence of 54.9% (43.4-66.0%). Eimeria quenda was less prevalent, at 7.0% (3.1-15.5%). The novel species E. angustus was present in 45.1% of sampled quenda (34.0-56.6%). A second novel morphotype of Eimeria was present in 2.8% of sampled quenda (0.9-9.7%). Mixed Eimeria spp. infections were present in 21/71 quenda (29.6%, 95% CI 20.2-41.1%). Molecular phylogenetic analyses of E. kanyana and E. angustus were conducted at the 18S rRNA and mitochondrial cytochrome oxidase loci. At both loci, two isolates identified as E. kanyana grouped in a phylogenetic clade with E. trichosuri. Five isolates identified as the novel E. angustus were most closely related to E. tropidura at the 18S locus. At the COI locus, no sequence data were available for E. tropidura; isolates of E. angustus grouped with E. sciurorum. Copyright © 2016 Elsevier Inc. All rights reserved.

  20. Immune response to rabbit coccidiosis: a comparison between infections with Eimeria flavescens and E. intestinalis

    Czech Academy of Sciences Publication Activity Database

    Pakandl, Michal; Hlásková, Lenka; Poplštein, M.; Nevečeřalová, M.; Vodička, T.; Salát, Jiří; Mucksová, J.

    2008-01-01

    Roč. 55, č. 1 (2008), s. 1-6 ISSN 0015-5683 R&D Projects: GA ČR GA524/05/2328 Institutional research plan: CEZ:AV0Z60220518 Keywords : rabbit coccidiosis * Eimeria intestinalis * Eimeria flavescens * immune response * ELISA * lymph ocyte proliferation * intraepithelial lymph ocytes Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 1.307, year: 2008

  1. Transcript and protein expression profile of PF11_0394, a Plasmodium falciparum protein expressed in salivary gland sporozoites

    Directory of Open Access Journals (Sweden)

    Schlarman Maggie S

    2012-03-01

    Full Text Available Abstract Background Plasmodium falciparum malaria is a significant problem around the world today, thus there is still a need for new control methods to be developed. Because the sporozoite displays dual infectivity for both the mosquito salivary glands and vertebrate host tissue, it is a good target for vaccine development. Methods The P. falciparum gene, PF11_0394, was chosen as a candidate for study due to its potential role in the invasion of host tissues. This gene, which was selected using a data mining approach from PlasmoDB, is expressed both at the transcriptional and protein levels in sporozoites and likely encodes a putative surface protein. Using reverse transcription-polymerase chain reaction (RT-PCR and green fluorescent protein (GFP-trafficking studies, a transcript and protein expression profile of PF11_0394 was determined. Results The PF11_0394 protein has orthologs in other Plasmodium species and Apicomplexans, but none outside of the group Apicomplexa. PF11_0394 transcript was found to be present during both the sporozoite and erythrocytic stages of the parasite life cycle, but no transcript was detected during axenic exoerythrocytic stages. Despite the presence of transcript throughout several life cycle stages, the PF11_0394 protein was only detected in salivary gland sporozoites. Conclusions PF11_0394 appears to be a protein uniquely detected in salivary gland sporozoites. Even though a specific function of PF11_0394 has not been determined in P. falciparum biology, it could be another candidate for a new vaccine.

  2. Genome-wide association study and biological pathway analysis of the Eimeria maxima response in broilers.

    Science.gov (United States)

    Hamzić, Edin; Buitenhuis, Bart; Hérault, Frédéric; Hawken, Rachel; Abrahamsen, Mitchel S; Servin, Bertrand; Elsen, Jean-Michel; Pinard-van der Laan, Marie-Hélène; Bed'Hom, Bertrand

    2015-11-25

    Coccidiosis is the most common and costly disease in the poultry industry and is caused by protozoans of the Eimeria genus. The current control of coccidiosis, based on the use of anticoccidial drugs and vaccination, faces serious obstacles such as drug resistance and the high costs for the development of efficient vaccines, respectively. Therefore, the current control programs must be expanded with complementary approaches such as the use of genetics to improve the host response to Eimeria infections. Recently, we have performed a large-scale challenge study on Cobb500 broilers using E. maxima for which we investigated variability among animals in response to the challenge. As a follow-up to this challenge study, we performed a genome-wide association study (GWAS) to identify genomic regions underlying variability of the measured traits in the response to Eimeria maxima in broilers. Furthermore, we conducted a post-GWAS functional analysis to increase our biological understanding of the underlying response to Eimeria maxima challenge. In total, we identified 22 single nucleotide polymorphisms (SNPs) with q value Eimeria maxima in broilers. Furthermore, the post-GWAS functional analysis indicates that biological pathways and networks involved in tissue proliferation and repair along with the primary innate immune response may play the most important role during the early stage of Eimeria maxima infection in broilers.

  3. Host specificity of turkey and chicken Eimeria: controlled cross-transmission studies and a phylogenetic view.

    Science.gov (United States)

    Vrba, Vladimir; Pakandl, Michal

    2015-03-15

    Protozoan parasites of the Eimeria genus have undergone extensive speciation and are now represented by a myriad of species that are specialised to different hosts. These species are highly host-specific and usually parasitise single host species, with only few reported exceptions. Doubts regarding the strict host specificity were frequent in the original literature describing coccidia parasitising domestic turkeys. The availability of pure characterised lines of turkey and chicken Eimeria species along with the recently developed quantitative PCR identification of these species allowed to investigate the issue of host specificity using well-controlled cross-transmission experiments. Seven species of gallinaceous birds (Gallus gallus, Meleagris gallopavo, Alectoris rufa, Perdix perdix, Phasianus colchicus, Numida meleagris and Colinus virginianus) were inoculated with six species and strains of turkey Eimeria and six species of chicken coccidia and production of oocysts was monitored. Turkey Eimeria species E. dispersa, E. innocua and E. meleagridis could complete their development in the hosts from different genera or even different families. Comparison of phylogenetic positions of these Eimeria species according to 18S rDNA and COI showed that the phylogeny cannot explain the observed patterns of host specificity. These findings suggest that the adaptation of Eimeria parasites to foreign hosts is possible and might play a significant role in the evolution and diversification of this genus. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Towards identifying novel anti-Eimeria agents: trace elements, vitamins, and plant-based natural products.

    Science.gov (United States)

    Wunderlich, Frank; Al-Quraishy, Saleh; Steinbrenner, Holger; Sies, Helmut; Dkhil, Mohamed A

    2014-10-01

    Eimeriosis, a widespread infectious disease of livestock, is caused by coccidian protozoans of the genus Eimeria. These obligate intracellular parasites strike the digestive tract of their hosts and give rise to enormous economic losses, particularly in poultry, ruminants including cattle, and rabbit farming. Vaccination, though a rational prophylactic measure, has not yet been as successful as initially thought. Numerous broad-spectrum anti-coccidial drugs are currently in use for treatment and prophylactic control of eimeriosis. However, increasing concerns about parasite resistance, consumer health, and environmental safety of the commercial drugs warrant efforts to search for novel agents with anti-Eimeria activity. This review summarizes current approaches to prevent and treat eimeriosis such as vaccination and commercial drugs, as well as recent attempts to use dietary antioxidants as novel anti-Eimeria agents. In particular, the trace elements selenium and zinc, the vitamins A and E, and natural products extracted from garlic, barberry, pomegranate, sweet wormwood, and other plants are discussed. Several of these novel anti-Eimeria agents exhibit a protective role against oxidative stress that occurs not only in the intestine of Eimeria-infected animals, but also in their non-parasitized tissues, in particular, in the first-pass organ liver. Currently, it appears to be promising to identify safe combinations of low-cost natural products with high anti-Eimeria efficacy for a potential use as feed supplementation in animal farming.

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

    Directory of Open Access Journals (Sweden)

    Hong Chen

    2013-01-01

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

  6. Comparative Plasmodium gene overexpression reveals distinct perturbation of sporozoite transmission by profilin.

    Science.gov (United States)

    Sato, Yuko; Hliscs, Marion; Dunst, Josefine; Goosmann, Christian; Brinkmann, Volker; Montagna, Georgina N; Matuschewski, Kai

    2016-07-15

    Plasmodium relies on actin-based motility to migrate from the site of infection and invade target cells. Using a substrate-dependent gliding locomotion, sporozoites are able to move at fast speed (1-3 μm/s). This motility relies on a minimal set of actin regulatory proteins and occurs in the absence of detectable filamentous actin (F-actin). Here we report an overexpression strategy to investigate whether perturbations of F-actin steady-state levels affect gliding locomotion and host invasion. We selected two vital Plasmodium berghei G-actin-binding proteins, C-CAP and profilin, in combination with three stage-specific promoters and mapped the phenotypes afforded by overexpression in all three extracellular motile stages. We show that in merozoites and ookinetes, additional expression does not impair life cycle progression. In marked contrast, overexpression of C-CAP and profilin in sporozoites impairs circular gliding motility and salivary gland invasion. The propensity for productive motility correlates with actin accumulation at the parasite tip, as revealed by combinations of an actin-stabilizing drug and transgenic parasites. Strong expression of profilin, but not C-CAP, resulted in complete life cycle arrest. Comparative overexpression is an alternative experimental genetic strategy to study essential genes and reveals effects of regulatory imbalances that are not uncovered from deletion-mutant phenotyping. © 2016 Sato et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  7. Eimeria atlapetesi nom. nov., a replacement name for Eimeria pileata Soriano-Vargas et al., 2015 (Apicomplexa: Eimeriidae), preoccupied by Eimeria pileata Straneva and Kelley, 1979 (Apicomplexa: Eimeriidae), with observations on histopathology and phylogenetic analysis.

    Science.gov (United States)

    Soriano-Vargas, Edgardo; Salgado-Miranda, Celene; Zepeda-Velázquez, Andrea Paloma; Medina, Juan Pablo; Janczur, Mariusz Krzysztof; González-Gómez, Maricruz; Flores-Valle, Izanami Tereira; Berto, Bruno Pereira; Lopes, Carlos Wilson Gomes

    2017-01-31

    Eimeria pileata Soriano-Vargas, Medina, Salgado-Miranda, García-Conejo, Galindo-Sánchez, Janczur, Berto and Lopes, 2015 is a junior homonym of Eimeria pileata Straneva and Kelley, 1979 and needs to be replaced. This coccidium was described from a rufous-capped brush finch Atlapetes pileatus Wagler in the Nevado de Toluca Natural Protected Area, Mexico. Thus, to maintain the original intent of the specific epithet derived from the scientific name of the type-host, the name Eimeria atlapetesi nom. nov. is proposed as a replacement name. Additionally, the current work reports another rufous-capped brush finch A. pileatus parasitized by E. atlapetesi in co-infection with an Isospora sp., providing observations of histopathology and phylogenetic analysis of 18S ribosomal RNA (rRNA) gene from E. atlapetesi. Endogenous forms of E. atlapetesi and Isospora sp. were observed in intestinal sections. Few oocysts of Isospora sp. were observed; therefore they were not morphologically or molecularly identified. In return, E. atlapetesi was identified and it was phylogenetically close to Eimeria dispersa Tyzzer, 1929 from the domestic turkey Meleagris gallopavo Linnaeus.

  8. Interleukin-10 neutralizing antibody for detection of intestinal luminal levels and as a dietary additive in Eimeria challenged broiler chicks.

    Science.gov (United States)

    Arendt, Maria K; Sand, Jordan M; Marcone, Taylor M; Cook, Mark E

    2016-02-01

    Interleukin-10 (IL-10) mRNA levels are increased within intestinal mucosa after Eimeria infection. IL-10 apical receptor presence on enterocytes suggests IL-10 is secreted into the intestinal lumen. Increased IL-10 has been shown to be central to the pathogenesis of numerous intracellular pathogens; we hypothesize luminal secretion of IL-10 enables Eimeria spp. infection in chickens. This study examines intestine luminal IL-10 levels and performance in broilers challenged with Eimeria when fed an anti-IL-10 antibody. Chicks were fed a diet (1 to 21 d) with control or anti-IL-10 antibody (0.34 g egg yolk antibody powder/Kg diet) with a saline or 10× dose of Advent coccidiosis vaccine on d 3. One chick per pen was euthanized on days 2, 4, 7, 10, 13, 16, and 19 post-challenge, bled, and intestines were collected for luminal fluid IL-10 concentrations. Body weight and feed intake were measured on d 21, and oocyst shedding was assessed on d 7 post-challenge. A significant Eimeria × antibody interaction on d 21 body weight (P < 0.05) showed chicks fed control antibody, but not anti-IL-10, had significant reductions in body weight when challenged with Eimeria spp. Oocyst shedding was increased with Eimeria challenge, but dietary antibody had no effect. Plasma carotenoid levels were reduced in Eimeria challenged chicks 4, 7, 10, and 16 days post-challenge compared to unchallenged chicks. Lack of an Eimeria × antibody interaction showed anti-IL-10 was not protective against Eimeria-induced decreases in plasma carotenoids. Eimeria challenge increased intestine luminal IL-10 on days 4 and 7 post-challenge in the cecum and jejunum, respectively, compared to unchallenged. Dietary anti-IL-10 decreased luminal IL-10 in the ileum on day 2 post-challenge when compared to control antibody fed chicks. No interaction between Eimeria challenge and antibody was observed on intestine luminal contents of IL-10, suggesting anti-IL-10 was ineffective at preventing increased Eimeria

  9. Identification of a major rif transcript common to gametocytes and sporozoites of Plasmodium falciparum

    DEFF Research Database (Denmark)

    Wang, Christian W; Mwakalinga, Steven B; Sutherland, Colin J

    2010-01-01

    ABSTRACT: BACKGROUND: The Plasmodium falciparum parasite is transmitted in its sexual gametocyte stage from man to mosquito and as asexual sporozoites from mosquito to man. Developing gametocytes sequester preferentially in the bone marrow, but mature stage gametocytes are released...

  10. Characterization of sporozoite surface antigens of Plasmodium falciparum, using monoclonal antibodies. Part of a coordinated programme on the preparation of irradiated vaccines against some human diseases

    International Nuclear Information System (INIS)

    Groot, M.

    1982-10-01

    Sporozoites are considered as a source of potential vaccine. Characterization of their antigens is therefore important and can be achieved by monoclonal antibodies. The purpose of this project is to study the production of monoclonal antibodies against sporozoites of P. falciparum. Various infections of mosquitoes were carried out during the period 1981-1982 to obtain antigens for the production of hybridomas. Hybridomas were produced from mice immunized through the bites of infected mosquitoes and by intravenous inoculation. The anti-sporozoite activity of the hybridomas was tested by an immunofluorescent antibody test using P. falciparum sporozoites as antigens. Positive immunofluorescence was seen in hybridoma cell lines tested with P. falciparum, whereas negative results were obtained when the cell lines were cross-reacted with other human species (P. vivax) and with a rodent malaria parasite (P. berghei)

  11. Biopathologic Characterization of Three Mixed Poultry Eimeria spp. Isolates

    Directory of Open Access Journals (Sweden)

    B Mansoori

    2011-09-01

    Full Text Available Background: Coccidiosis of domestic fowl, caused by species of the Genus Eimeria, is responsi­ble for important economic losses in poultry production. Because different species and/or strains can vary in pathogenicity and other biological parameters, their precise characteriza­tion is important for epizootiological studies.Methods: Fifty samples from litter, whole intestinal tract and feces were collected from poul­try houses located in different provinces of Iran. One hundred twenty male day-old broi­ler chicks were challenged with three selected isolates. Data on weight gain, Food Conversion Ratio (FCR, food intake, lesion scoring and shedding of oocysts per gram of feces were rec­orded and analyzed by the Duncan's test.Results: In all treatments, the challenged groups had statistically significant lower weight gain than that of unchallenged control group. Isolate three caused the lowest weight gain and food intake and the worst lesion score as well as FCR. Despite originating from close geographi­cal regions for isolates 1 and 2, the difference in biopathologic factors may be either due to different proportion of identified species or the different pathogenicity of the species present in the isolates.Conclusion: The results highlight the importance of considering various species of Eimeria in designing the preventive, control and treatment strategies to prevent coccidiosis in different regions of Iran. Further characterization of each isolate would be the next step to provide a basis for coccidiosis research with well-characterized local isolates.

  12. Blood Profile of Rabbits Infected with Eimeria magna

    Directory of Open Access Journals (Sweden)

    A Hana

    2011-09-01

    Full Text Available Abstract. The research aimed at determining the blood profile of local rabbits infected with different dose of Eimeria magna oocysts. This research used 45 male rabbits with the age of 4 month old, range from 1.5 to 1.8 kg, clinically healthy and free from coccidiosis. The rabbits were randomly divided into 3 groups, group I as control (K-0 was given 1.0 ml distilled water/rabbit orally, group II (K-10 was infected with single dose of 10x106 oocysts of E. magna/rabbit orally, and group III (K-20 was infected with single dose of 20x106 oocysts of E. magna/rabbit orally. After infection, rabbits were examined for clinical signs, body weight and temperature daily for five days. Blood samples were drawn from the vena marginalis to examine the number of erythrocytes, hemoglobine, packed cell volume (PCV, leukocytes and its deferent, total protein plasma (TPP and fibrinogen, activities of alkaline phosphatase (ALP, alanine amino transferase (ALT, and aspartat aminotransferase (AST. The data were statistically analyzed by two-way anova using factorial design. The results of this research showed that the infection of E. magna in rabbits caused fever and weight loss, accompanied by normochromic microcytic anemia (at doses of 10x106 oocysts, macrocytic normochromic (at doses of 20x106 oocysts, leukocytosis, lymphocytosis, hiperfibrinogenemia, and increased of ALP activity. There were correlations between clinical symptoms and blood profile of rabbits infected with E. magna for five days. The higher the dose and the longer the infection of E. magna in rabbits caused weight loss, increased body temperature, MCV (microcytic to macrocytic, leukocyte, fibrinogen and ALP activity. These findings were useful to have a better understanding of pathophysiology of E. magna infection in  rabbits. Key Words: Eimeria magna, oocyst, rabbit, blood profile A Hana et al/Animal Production 13(3:185-190 (2011

  13. Comparative transcriptome analysis of second- and third-generation merozoites of Eimeria necatrix.

    Science.gov (United States)

    Su, Shijie; Hou, Zhaofeng; Liu, Dandan; Jia, Chuanli; Wang, Lele; Xu, Jinjun; Tao, Jianping

    2017-08-16

    Eimeria is a common genus of apicomplexan parasites that infect diverse vertebrates, most notably poultry, causing serious disease and economic losses. Eimeria species have complex life-cycles consisting of three developmental stages. However, the molecular basis of the Eimeria reproductive mode switch remains an enigma. Total RNA extracted from second- (MZ-2) and third-generation merozoites (MZ-3) of Eimeria necatrix was subjected to transcriptome analysis using RNA sequencing (RNA-seq) followed by qRT-PCR validation. A total of 6977 and 6901 unigenes were obtained from MZ-2 and MZ-3, respectively. Approximately 2053 genes were differentially expressed genes (DEGs) between MZ-2 and MZ-3. Compared with MZ-2, 837 genes were upregulated and 1216 genes were downregulated in MZ-3. Approximately 95 genes in MZ-2 and 48 genes in MZ-3 were further identified to have stage-specific expression. Gene ontology category and KEGG analysis suggested that 216 upregulated genes in MZ-2 were annotated by 70 GO assignments, 242 upregulated genes were associated with 188 signal pathways, while 321 upregulated genes in MZ-3 were annotated by 56 GO assignments, 322 upregulated genes were associated with 168 signal pathways. The molecular functions of upregulated genes in MZ-2 were mainly enriched for protein degradation and amino acid metabolism. The molecular functions of upregulated genes in MZ-3 were mainly enriched for transcriptional activity, cell proliferation and cell differentiation. To the best of our knowledge, this is the first RNA-seq data study of the MZ-2 and MZ-3 stages of E. necatrix; it demonstrates a high number of differentially expressed genes between the MZ-2 and MZ-3 of E. necatrix. This study forms a basis for deciphering the molecular mechanisms underlying the shift from the second to third generation schizogony in Eimeria. It also provides valuable resources for future studies on Eimeria, and provides insight into the understanding of reproductive mode

  14. Adenovirus Particles that Display the Plasmodium falciparum Circumsporozoite Protein NANP Repeat Induce Sporozoite-Neutralizing Antibodies in Mice

    Science.gov (United States)

    Palma, Christopher; Overstreet, Michael G.; Guedon, Jean-Marc; Hoiczyk, Egbert; Ward, Cameron; Karen, Kasey A.; Zavala, Fidel; Ketner, Gary

    2011-01-01

    Adenovirus particles can be engineered to display exogenous peptides on their surfaces by modification of viral capsid proteins, and particles that display pathogen-derived peptides can induce protective immunity. We constructed viable recombinant adenoviruses that display B-cell epitopes from the Plasmodium falciparum circumsporozoite protein (PfCSP) in the major adenovirus capsid protein, hexon. Recombinants induced high-titer antibodies against CSP when injected intraperitoneally into mice. Serum obtained from immunized mice recognized both recombinant PfCSP protein and P. falciparum sporozoites, and neutralized P. falciparum sporozoites in vitro. Replicating adenovirus vaccines have provided economical protection against adenovirus disease for over three decades. The recombinants described here may provide a path to an affordable malaria vaccine in the developing world. PMID:21199707

  15. Detection and molecular characterization of Cryptosporidium and Eimeria species in Philippine bats.

    Science.gov (United States)

    Murakoshi, Fumi; Recuenco, Frances C; Omatsu, Tsutomu; Sano, Kaori; Taniguchi, Satoshi; Masangkay, Joseph S; Alviola, Philip; Eres, Eduardo; Cosico, Edison; Alvarez, James; Une, Yumi; Kyuwa, Shigeru; Sugiura, Yuki; Kato, Kentaro

    2016-05-01

    The genus Cryptosporidium, which is an obligate intracellular parasite, infects various vertebrates and causes a diarrheal disease known as cryptosporidiosis. Bats are naturally infected with zoonotic pathogens; thus, they are potential reservoirs of parasites. We investigated the species and genotype distribution as well as prevalence of Cryptosporidium and Eimeria in Philippine bats. We captured and examined 45 bats; four were positive for Cryptosporidium spp. and seven were positive for Eimeria spp. We detected Cryptosporidium bat genotype II from Ptenochirus jagori. Three other Cryptosporidium sequences, detected from Rhinolophus inops, Cynopterus brachyotis, and Eonycteris spelaea, could not be classified as any known species or genotype; we therefore propose the novel genotype Cryptosporidium bat genotypes V, VI, and VII. Bat genotype V is associated with human cryptosporidiosis clade, and therefore, this genotype may be transmissible to humans. Among the Eimeria sequences, BE3 detected from Scotophilus kuhlii was classified with known bat and rodent clades; however, other sequences detected from C. brachyotis, E. spelaea, Rousettus amplexicaudatus, and R. inops could not be classified with known Eimeria species. These isolates might represent a new genotype. Our findings demonstrate that the bats of the Philippines represent a reservoir of multiple Cryptosporidium and Eimeria spp.

  16. The ETRAMP family member SEP2 is expressed throughout Plasmodium berghei life cycle and is released during sporozoite gliding motility.

    Science.gov (United States)

    Currà, Chiara; Di Luca, Marco; Picci, Leonardo; de Sousa Silva Gomes dos Santos, Carina; Siden-Kiamos, Inga; Pace, Tomasino; Ponzi, Marta

    2013-01-01

    The early transcribed membrane proteins ETRAMPs belong to a family of small, transmembrane molecules unique to Plasmodium parasite, which share a signal peptide followed by a short lysine-rich stretch, a transmembrane domain and a variable, highly charged C-terminal region. ETRAMPs are usually expressed in a stage-specific manner. In the blood stages they localize to the parasitophorous vacuole membrane and, in described cases, to vesicle-like structures exported to the host erythrocyte cytosol. Two family members of the rodent parasite Plasmodium berghei, uis3 and uis4, localize to secretory organelles of sporozoites and to the parasitophorous membrane vacuole of the liver stages. By the use of specific antibodies and the generation of transgenic lines, we showed that the P. berghei ETRAMP family member SEP2 is abundantly expressed in gametocytes as well as in mosquito and liver stages. In intracellular parasite stages, SEP2 is routed to the parasitophorous vacuole membrane while, in invasive ookinete and sporozoite stages, it localizes to the parasite surface. To date SEP2 is the only ETRAMP protein detected throughout the parasite life cycle. Furthermore, SEP2 is also released during gliding motility of salivary gland sporozoites. A limited number of proteins are known to be involved in this key function and the best characterized, the CSP and TRAP, are both promising transmission-blocking candidates. Our results suggest that ETRAMP members may be viewed as new potential candidates for malaria control.

  17. Re-assessing the relationship between sporozoite dose and incubation period in Plasmodium vivax malaria: a systematic re-analysis.

    Science.gov (United States)

    Lover, Andrew A; Coker, Richard J

    2014-05-01

    Infections with the malaria parasite Plasmodium vivax are noteworthy for potentially very long incubation periods (6-9 months), which present a major barrier to disease elimination. Increased sporozoite challenge has been reported to be associated with both shorter incubation and pre-patent periods in a range of human challenge studies. However, this evidence base has scant empirical foundation, as these historical analyses were limited by available analytic methods, and provides no quantitative estimates of effect size. Following a comprehensive literature search, we re-analysed all identified studies using survival and/or logistic models plus contingency tables. We have found very weak evidence for dose-dependence at entomologically plausible inocula levels. These results strongly suggest that sporozoite dosage is not an important driver of long-latency. Evidence presented suggests that parasite strain and vector species have quantitatively greater impacts, and the potential existence of a dose threshold for human dose-response to sporozoites. Greater consideration of the complex interplay between these aspects of vectors and parasites are important for human challenge experiments, vaccine trials, and epidemiology towards global malaria elimination.

  18. Longitudinal prevalence, oocyst shedding and molecular characterisation of Eimeria species in sheep across four states in Australia.

    Science.gov (United States)

    Yang, Rongchang; Jacobson, Caroline; Gardner, Graham; Carmichael, Ian; Campbell, Angus J D; Ryan, Una

    2014-10-01

    The prevalence of Eimeria in sheep in Australia has not been well described, therefore a quantitative PCR (qPCR) was developed, validated and used to study the prevalence and oocyst concentration in lamb faecal samples at three sampling periods (weaning, post-weaning and pre-slaughter) from eight farms across South Australia, New South Wales, Victoria and Western Australia. A total of 3412 faecal samples were collected from approximately 1182 lambs across the 4 states and screened for the presence of Eimeria using this qPCR at the 18S ribosomal RNA (rRNA) locus. A subset of positives was typed by sequence analysis at the 18S locus. The overall prevalence was 18.1% (95% CI 16.8-19.3%) and of the 616 positives, 118 were successfully genotyped. The prevalence of Eimeria was highest in NSW and peaked at 70% during the post-weaning period. The range of oocyst shedding per gram of faeces (g(-1)) at weaning, post-weaning and pre-slaughter overall across all states was 23-2.1×10(7), 23-1.3×10(7) and 23-2.1×10(5), respectively. Median Eimeria shedding g(-1) was higher during post-weaning (1.1×10(3)) and pre-slaughter (1.1×10(3)) than during weaning (206). The following species were identified: Eimeria crandallis, Eimeria ahsata, Eimeria ovinoidalis, Eimeria weybridgensis and Eimeria cylindrica. Of these, E. crandallis and E. ovinoidalis, the most pathogenic species in sheep were responsible for 58.5% of infections typed. This highlights a need for further research to quantify the production impacts of Eimeria in sheep. Copyright © 2014 Elsevier Inc. All rights reserved.

  19. Simultaneous identification and DNA barcoding of six Eimeria species infecting turkeys using PCR primers targeting the mitochondrial cytochrome c oxidase subunit I (mtCOI) locus.

    Science.gov (United States)

    Hafeez, Mian A; Shivaramaiah, Srichaitanya; Dorsey, Kristi Moore; Ogedengbe, Mosun E; El-Sherry, Shiem; Whale, Julia; Cobean, Julie; Barta, John R

    2015-05-01

    Species-specific PCR primers targeting the mitochondrial cytochrome c oxidase subunit I (mtCOI) locus were generated that allow for the specific identification of the most common Eimeria species infecting turkeys (i.e., Eimeria adenoeides, Eimeria meleagrimitis, Eimeria gallopavonis, Eimeria meleagridis, Eimeria dispersa, and Eimeria innocua). PCR reaction chemistries were optimized with respect to divalent cation (MgCl2) and dNTP concentrations, as well as PCR cycling conditions (particularly anneal temperature for primers). Genomic DNA samples from single oocyst-derived lines of six Eimeria species were tested to establish specificity and sensitivity of these newly designed primer pairs. A mixed 60-ng total DNA sample containing 10 ng of each of the six Eimeria species was used as DNA template to demonstrate specific amplification of the correct product using each of the species-specific primer pairs. Ten nanograms of each of the five non-target Eimeria species was pooled to provide a non-target, control DNA sample suitable to test the specificity of each primer pair. The amplifications of the COI region with species-specific primer pairs from pooled samples yielded products of expected sizes (209 to 1,012 bp) and no amplification of non-target Eimeria sp. DNA was detected using the non-target, control DNA samples. These primer pairs specific for Eimeria spp. of turkeys did not amplify any of the seven Eimeria species infecting chickens. The newly developed PCR primers can be used as a diagnostic tool capable of specifically identifying six turkey Eimeria species; additionally, sequencing of the PCR amplification products yields sequence-based genotyping data suitable for identification and molecular phylogenetics.

  20. The complete mitochondrial genome sequence of Eimeria innocua (Eimeriidae, Coccidia, Apicomplexa).

    Science.gov (United States)

    Hafeez, Mian Abdul; Vrba, Vladimir; Barta, John Robert

    2016-07-01

    The complete mitochondrial genome of Eimeria innocua KR strain (Eimeriidae, Coccidia, Apicomplexa) was sequenced. This coccidium infects turkeys (Meleagris gallopavo), Bobwhite quails (Colinus virginianus), and Grey partridges (Perdix perdix). Genome organization and gene contents were comparable with other Eimeria spp. infecting galliform birds. The circular-mapping mt genome of E. innocua is 6247 bp in length with three protein-coding genes (cox1, cox3, and cytb), 19 gene fragments encoding large subunit (LSU) rRNA and 14 gene fragments encoding small subunit (SSU) rRNA. Like other Apicomplexa, no tRNA was encoded. The mitochondrial genome of E. innocua confirms its close phylogenetic affinities to Eimeria dispersa.

  1. Zinc Supplementation against Eimeria acervulina-Induced Oxidative Damage in Broiler Chickens

    Directory of Open Access Journals (Sweden)

    Nedyalka V. Georgieva

    2011-01-01

    Full Text Available This study was undertaken to determine the dietary supplements of Zn containing diet on the antioxidant status in chickens experimentally infected with Eimeria acervulina. The antioxidant status was monitored via determination of MDA concentrations and erythrocyte SOD and CAT activities, as well as vitamin E, vitamin C, Cu, and Zn in liver, muscle, and serum. The results showed increased MDA (<.05, CAT (<.001, and decreased SOD (<.001 in the infected birds. Significant changes in Cu and Zn concentrations and dramatically reduction of vitamin C and E concentrations in the infected chickens were found. The observed deviations in the studied enzymes and nonenzymatic parameters evidence the occurrence of oxidative stress following the infection and impaired antioxidant status of chickens, infected with Eimeria acervulina. Our results proved the ameliorating role of CuZn(OH3Cl (0.170 g per kg food against Eimeria acervulina-induced oxidative damage in infected chickens.

  2. Mechanisms of invasion from sporozoite and merozoíto of Plasmodium

    Directory of Open Access Journals (Sweden)

    Lilian M. Spencer

    2016-05-01

    Full Text Available Malaria or paludismo is caused in humans by four species of Plasmodium belonging to phylum Apicomplexa: ovale, malaria, vivax and falciparum, being the last, the responsible of the clinical complication and death in the vertebrate host. Plasmodium parasite possess a specialized secretory organelles called rhoptries, micronemes and dense granules that facilitate invasion of host cells. The sporozoite stage of Plasmodium travels through the different cells of vertebrate host until it reaches the hepatocyte and have been form the parasitophorous vacuole. The infected hepatocytes rupture, results in the releasing thousands of daughter merozoites that invade the erythrocytes with the formation of parasitophorous vacuole too. Several researchers suggest the gliding motility mechanism as the responsible of hepatocyte invasion. While, which the erythrocyte invasion process has been described as the result of tree steps: first contact, re-orientation and invasion. In this review the surface proteins of merozoites and esporozoites are pointed out as the most important factors for the molecular invasion mechanisms until the elaboration of the parasitophorous vacuole. These proteins that take part in these mechanisms are the possible candidates in the design of an anti-malaria vaccine.

  3. Interrogating the Plasmodium Sporozoite Surface: Identification of Surface-Exposed Proteins and Demonstration of Glycosylation on CSP and TRAP by Mass Spectrometry-Based Proteomics.

    Directory of Open Access Journals (Sweden)

    Kristian E Swearingen

    2016-04-01

    Full Text Available Malaria parasite infection is initiated by the mosquito-transmitted sporozoite stage, a highly motile invasive cell that targets hepatocytes in the liver for infection. A promising approach to developing a malaria vaccine is the use of proteins located on the sporozoite surface as antigens to elicit humoral immune responses that prevent the establishment of infection. Very little of the P. falciparum genome has been considered as potential vaccine targets, and candidate vaccines have been almost exclusively based on single antigens, generating the need for novel target identification. The most advanced malaria vaccine to date, RTS,S, a subunit vaccine consisting of a portion of the major surface protein circumsporozoite protein (CSP, conferred limited protection in Phase III trials, falling short of community-established vaccine efficacy goals. In striking contrast to the limited protection seen in current vaccine trials, sterilizing immunity can be achieved by immunization with radiation-attenuated sporozoites, suggesting that more potent protection may be achievable with a multivalent protein vaccine. Here, we provide the most comprehensive analysis to date of proteins located on the surface of or secreted by Plasmodium falciparum salivary gland sporozoites. We used chemical labeling to isolate surface-exposed proteins on sporozoites and identified these proteins by mass spectrometry. We validated several of these targets and also provide evidence that components of the inner membrane complex are in fact surface-exposed and accessible to antibodies in live sporozoites. Finally, our mass spectrometry data provide the first direct evidence that the Plasmodium surface proteins CSP and TRAP are glycosylated in sporozoites, a finding that could impact the selection of vaccine antigens.

  4. Prevalence and risk factors associated to Eimeria spp. infection in unweaned alpacas (Vicugna pacos) from Southern Peru.

    Science.gov (United States)

    Díaz, Pablo; Panadero, Rosario; López, Rosalía; Cordero, Aida; Pérez-Creo, Ana; López, Ceferino M; Fernández, Gonzalo; Díez-Baños, Pablo; Morrondo, Patrocinio

    2016-01-01

    A total of 350 faecal samples from unweaned alpacas over 3 months of age were collected from 23 herds in order to determine the prevalence of Eimeria spp. in Southern Peru and to identify the risk factors associated to Eimeria infection in young alpacas. Samples were examined by a flotation technique and the identification of risk factors was assessed by a logistic regression analysis. Sixty four percent of the examined animals shed Eimeria oocysts; herd prevalence was 96%, with an intra-herd prevalence of 60% (range 5.9-100%). Five different Eimeria species were identified, being E. lamae (91%), E. alpacae (87%) and E. punoensis (78%) the most prevalent; E. macusaniensis (35%) and E. ivitaensis (13%) were less common. Mixed-species infections were more frequent (78%) than single infections (22%). E. lamae was the most common monospecific infection and E. lamae/E. alpacae the most frequent association. The geographical area has a significant effect on Eimeria infection rates (74.9% wet Puna vs 37.4% dry Puna) as well as the breeding system (65.1% traditional vs 63.8% modern). In contrast, the sex of the animals (64.6% males vs 64.0% females) showed no influence on the prevalence of infection by Eimeria. The high prevalence found at both individual and herd level and the common presence of highly pathogenic Eimeria species may lead to important economic losses for alpaca breeders and could require the implementation of suitable control measures.

  5. An analysis of the spatial distribution of Plasmodium sporozoites and effects of climatic correlates on malaria infection in Anyigba town, Nigeria.

    Science.gov (United States)

    Ifatimehin, Olarewaju Oluseyi; Falola, O O; Odogbo, E V

    2013-10-27

    The infectivity of sporozoites on both mosquitoes and human is the major cause of malaria infection on its host, Man. Malaria infection had continued to blossom despite measures to curb it. Clinically diagnosed malaria data for 3 years, capture of mosquitoes for laboratory analysis to determining the infectivity of sporozoites, responses from the population on the number of episode of malaria in the last 60 days were all collected and generated, and also subjected to various analysis using methods accepted tools and methods. A fifteen weeks climatic data was also collected. It was discovered that malaria incidence of 467.2853/1000 persons is very high. This high rate is possible as out of every 10 mosquitoes in Anyigba, 4 are infected by sporozoites and can possibly transmit these sporozoites during blood feeding on the population. This is affirmed by the prevalence of malaria by 54.75% among Anyigba's population. At p>001 (0.829), climatic variables and sporozoites rate showed a strong affinity with the prevalence of malaria. The risk map showed that the university community and the surrounding students' lodges are areas of very high risk. Therefore, the populace is strongly advised to employed practicable measures such as regular environmental sanitation and the use of Insecticidal Treated Nets (ITN) in order to drastically address this epidemic.

  6. Anti-Eimeria activity of berberine and identification of associated gene expression changes in the mouse jejunum infected with Eimeria papillata.

    Science.gov (United States)

    Dkhil, Mohamed A; Metwaly, Mahmoud S; Al-Quraishy, Saleh; Sherif, Nour E; Delic, Denis; Al Omar, Suliman Y; Wunderlich, Frank

    2015-04-01

    Plant-based natural products are promising sources for identifying novel agents with potential anti-Eimeria activity. This study explores possible effects of berberine on Eimeria papillata infections in the jejunum of male Swiss albino mice. Berberine chloride, when daily administered to mice during infection, impairs intracellular development and multiplication of E. papillata, evidenced as 60% reduction of maximal fecal output of oocysts on day 5 p.i. Concomitantly, berberine attenuates the inflammatory response, evidenced as decreased messenger RNA (mRNA) expression of IL-1β, IL-6, TNFα, IFNγ, and iNOS, as well as the oxidative stress response, evidenced as impaired increase in malondialdehyde, nitrate, and H2O2 and as prevented decrease in glutathione and catalase activity. Berberine also alters gene expression in the infected jejunum. On day 5 p.i., mRNA expression of 29 genes with annotated functions is more than 10-fold upregulated and that of 14 genes downregulated. Berberine downregulates the genes Xaf1, Itgb3bp, and Faim3 involved in apoptotic processes and upregulates genes involved in innate immune responses, as e.g., Colec11, Saa2, Klra8, Clec1b, and Crtam, especially the genes Cpa3, Fcer1a, and Mcpt1, Mcpt2, and Mcpt4 involved in mast cell activity. Additionally, 18 noncoding lincRNA species are differentially expressed more than 10-fold under berberine. Our data suggest that berberine induces hosts to exert anti-Eimeria activity by attenuating the inflammatory and oxidative stress response, by impairing apoptotic processes, and by activating local innate immune responses and epigenetic mechanisms in the host jejunum. Berberine has the potential as an anti-Eimeria food additive in animal farming.

  7. A new Eimeria (Apicomplexa: Eimeriidae), possessing mitra-shaped oocysts, from the Neotropical chelid turtle Batrachemys heliostemma (Testudines: Chelidae), and its comparison with Eimeria mitraria (Laveran & Mesnil 1902)

    Czech Academy of Sciences Publication Activity Database

    Široký, P.; Kamler, M.; Modrý, David

    2006-01-01

    Roč. 101, č. 5 (2006), s. 555-558 ISSN 0074-0276 R&D Projects: GA ČR GP524/03/D104; GA ČR GD524/03/H133 Institutional research plan: CEZ:AV0Z60220518 Keywords : Batrachemys * Eimeria Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 1.208, year: 2006

  8. Improved resistance to Eimeria acervulina infection in chickens due to dietary supplementation with garlic metabolites

    Science.gov (United States)

    The effects of a compound including secondary metabolites of garlic, propyl thiosulfinate (PTS) and propyl thiosulfinatate oxide (PTSO), on in vitro and in vivo parameters of chicken gut immunity during experimental Eimeria acervulina infection were evaluated. In in vitro assays, the compound of P...

  9. Eimeria Species and Genetic Background Influence the Serum Protein Profile of Broilers with Coccidiosis

    Science.gov (United States)

    Gilbert, Elizabeth R.; Cox, Chasity M.; Williams, Patricia M.; McElroy, Audrey P.; Dalloul, Rami A.; Ray, W. Keith; Barri, Adriana; Emmerson, Derek A.; Wong, Eric A.; Webb, Kenneth E.

    2011-01-01

    Background Coccidiosis is an intestinal disease caused by protozoal parasites of the genus Eimeria. Despite the advent of anti-coccidial drugs and vaccines, the disease continues to result in substantial annual economic losses to the poultry industry. There is still much unknown about the host response to infection and to date there are no reports of protein profiles in the blood of Eimeria-infected animals. The objective of this study was to evaluate the serum proteome of two genetic lines of broiler chickens after infection with one of three species of Eimeria. Methodology/Principal Findings Birds from lines A and B were either not infected or inoculated with sporulated oocysts from one of the three Eimeria strains at 15 d post-hatch. At 21 d (6 d post-infection), whole blood was collected and lesion scoring was performed. Serum was harvested and used for 2-dimensional gel electrophoresis. A total of 1,266 spots were quantitatively assessed by densitometry. Protein spots showing a significant effect of coccidia strain and/or broiler genetic line on density at PEimeria infection and in identifying molecular targets for diagnostic screening and development of alternative preventative and therapeutic methods. PMID:21297942

  10. Genome-Wide Association Study for Response to Eimeria maxima Challenge in Broilers

    DEFF Research Database (Denmark)

    Hamzic, Edin; Bed'hom, Bertrand; Hérault, Frédéric

    Use of genetic tools for improvement of host’s response is considered as a promising complementary approach for coccidiosis control. Therefore, we performed genome wide association study (GWAS) for response to Eimeria maxima challenge in broilers. The challenge was done on 2024 Cobb500 broilers. We...

  11. The genome of Eimeria falciformis--reduction and specialization in a single host apicomplexan parasite.

    Science.gov (United States)

    Heitlinger, Emanuel; Spork, Simone; Lucius, Richard; Dieterich, Christoph

    2014-08-20

    The phylum Apicomplexa comprises important unicellular human parasites such as Toxoplasma and Plasmodium. Eimeria is the largest and most diverse genus of apicomplexan parasites and some species of the genus are the causative agent of coccidiosis, a disease economically devastating in poultry. We report a complete genome sequence of the mouse parasite Eimeria falciformis. We assembled and annotated the genome sequence to study host-parasite interactions in this understudied genus in a model organism host. The genome of E. falciformis is 44 Mb in size and contains 5,879 predicted protein coding genes. Comparative analysis of E. falciformis with Toxoplasma gondii shows an emergence and diversification of gene families associated with motility and invasion mainly at the level of the Coccidia. Many rhoptry kinases, among them important virulence factors in T. gondii, are absent from the E. falciformis genome. Surface antigens are divergent between Eimeria species. Comparisons with T. gondii showed differences between genes involved in metabolism, N-glycan and GPI-anchor synthesis. E. falciformis possesses a reduced set of transmembrane transporters and we suggest an altered mode of iron uptake in the genus Eimeria. Reduced diversity of genes required for host-parasite interaction and transmembrane transport allow hypotheses on host adaptation and specialization of a single host parasite. The E. falciformis genome sequence sheds light on the evolution of the Coccidia and helps to identify determinants of host-parasite interaction critical for drug and vaccine development.

  12. Eimeria infection in calves under local farming system in and around ...

    African Journals Online (AJOL)

    High infection rates occur from environments that were already contaminated with infected animals. A study on the prevalence, species and risk of occurrence of Eimeria species in calves was conducted at Asella, Oromia Regional State, Ethiopia. Management systems, breed, age, sex, and site were considered as variables ...

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

    DEFF Research Database (Denmark)

    Sommer, C.

    1998-01-01

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

  14. Eimeria terraepokotorum sp.n. (Apicomplexa: Eimeriidae) from Hoplobatrachus occipitalis (Anura: Ranidae) from Kenya

    Czech Academy of Sciences Publication Activity Database

    Jirků, M.; Modrý, David

    2006-01-01

    Roč. 45, č. 4 (2006), s. 443-447 ISSN 0065-1583 R&D Projects: GA ČR GD524/03/H133; GA ČR GA206/03/1544 Institutional research plan: CEZ:AV0Z60220518 Keywords : Eimeria * coccidia * Anura Subject RIV: EG - Zoology Impact factor: 1.162, year: 2006

  15. Mucin gene mRNA levels in broilers challenged with eimeria and/or Clostridium perfringens.

    Science.gov (United States)

    Kitessa, Soressa M; Nattrass, Gregory S; Forder, Rebecca E A; McGrice, Hayley A; Wu, Shu-Biao; Hughes, Robert J

    2014-09-01

    The effects of Eimeria (EM) and Clostridium perfringens (CP) challenges on the mRNA levels of genes involved in mucin (Muc) synthesis (Muc2, Muc5ac, Muc13, and trefoil family factor-2 [TFF2]), inflammation (tumor necrosis factor alpha [TNF-alpha] and interleukin-18 [IL-18]), and metabolic processes (cluster of differentiation [CD]36) in the jejunum of broilers were investigated. Two parallel experiments involving 1) EM challenge and 2) EM and CP challenges were conducted. The first experiment was a 2 X 2 study with 12 birds per treatment (N = 48) involving fishmeal substitution (25%) in the diet (FM) and EM challenge. The treatments were: Control (FM-, EM-), Fishmeal (FM+, EM-), EM challenge (FM-, EM+), and fishmeal substitution and EM challenge (FM+, EM+). The second experiment was a 2 X 2 X 2 experiment with six birds per treatment (N = 48) involving fishmeal (FM-, FM+), Eimeria (EM-, EM+), and C perfringens (CP-, CP+). In both arms of the study, male broilers were given a starter diet for the whole period of 16 days, except those assigned to FM+, where 25% of the starter ration was replaced with fishmeal from days 8 to 14. EM inoculation was performed on day 9 and CP inoculation on days 14 and 15. The EM challenge birds were euthanatized for sampling on day 13; postmortem examination and sampling for the Eimeria plus C perfringens challenge arm of the study were on day 16. In the Eimeria challenge arm of the study, fishmeal supplementation significantly suppressed the mRNA levels of TNF-alpha, TFF2, and IL-18 pre-CP inoculation but simultaneously increased the levels of Muc13 and CD36 mRNAs. Birds challenged with Eimeria exhibited increased mRNA levels of Muc13, Muc5ac, TNF-alpha, and IL-18. In the Eimeria and C. perfringens challenge arm, birds exposed to EM challenge exhibited significantly lower mRNA levels of Muc2 and CD36. The mRNA levels of CD36 were also significantly suppressed by CP challenge. Our results showed that the transcription of mucin synthesis

  16. A new species of Eimeria (Apicomplexa: Eimeriidae) from Californian rabbits in Hebei Province, China.

    Science.gov (United States)

    Cui, Ping; Liu, Hongbin; Fang, Sufang; Gu, Xiaolong; Wang, Peng; Liu, Chunling; Tao, Geru; Liu, Xianyong; Suo, Xun

    2017-10-01

    Rabbit coccidiosis is caused by infection with one or usually several Eimeria species, parasitizing in hepatobiliary ducts or intestinal epithelium of rabbits. To date, 11 species of rabbit coccidia have been well documented. Here we report a new species of Eimeria from rabbits. Sporulated oocysts were ellipsoidal to slightly ovoidal, 37.4 (32.6-41.2) μm in length, 23.5 (20.9-25.5) μm in width, with a shape index (length/width) 1.6 (1.43-1.91) and smooth, bilayered, homogeneously thick wall. The micropyle was obvious and with an inner diameter of 6.2 (5.0-7.5) μm. Both oocyst residuum and polar granule were absent. Sporocysts were ellipsoidal to elongate, 17.2 (13.2-20.0) μm long and 8.4 (7.5-9.1) μm wide, with a shape index (length/width) of 2.1 (1.74-2.21) and the presence of Stieda body and sporocyst residuum. The prepatent period was 132h. Phylogenetic analysis showed that 18S rDNA sequence of the new species clustered together with the 11 rabbit Eimeria species into a clade. However, ITS-1 sequence of the new species shared low similarities (27.1%-30%) with those of 11 rabbit Eimeria species. As the data above supported the erection of a new species, we named it as Eimeria kongi n. sp., in honor of Fanyao Kong, a Chinese parasitologist. The finding of the new species has important implications for the diagnosis and prevention of rabbit coccidiosis. Copyright © 2017. Published by Elsevier B.V.

  17. 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.

  18. Genetic analysis of resistance to ticks, gastrointestinal nematodes and Eimeria spp. in Nellore cattle.

    Science.gov (United States)

    Passafaro, Tiago Luciano; Carrera, Juan Pablo Botero; dos Santos, Livia Loiola; Raidan, Fernanda Santos Silva; dos Santos, Dalinne Chrystian Carvalho; Cardoso, Eduardo Penteado; Leite, Romário Cerqueira; Toral, Fabio Luiz Buranelo

    2015-06-15

    The aim of the present study was to obtain genetic parameters for resistance to ticks, gastrointestinal nematodes (worms) and Eimeria spp. in Nellore cattle, analyze the inclusion of resistance traits in Nellore breeding programs and evaluate genetic selection as a complementary tool in parasite control programs. Counting of ticks, gastrointestinal nematode eggs and Eimeria spp. oocysts per gram of feces totaling 4270; 3872 and 3872 records from 1188; 1142 and 1142 animals, respectively, aged 146 to 597 days were used. The animals were classified as resistant (counts equal to zero) or susceptible (counts above zero) to each parasite. The statistical models included systematics effects of contemporary groups and the mean trajectory. The random effects included additive genetic effects, direct permanent environmental effects and residual. The mean trajectory and random effects were modeled with linear Legendre polynomials for all traits except for the mean trajectory of resistance to Eimeria spp., which employed the cubic polynomial. Heritability estimates were of low to moderate magnitude and ranged from 0.06 to 0.30, 0.06 to 0.33 and 0.04 to 0.33 for resistance to ticks, gastrointestinal nematodes and Eimeria spp., respectively. The posterior mean of genetic and environmental correlations for the same trait at different ages (205, 365, 450 and 550 days) were favorable at adjacent ages and unfavorable at distant ages. In general, the posterior mean of the genetic and environmental correlations between traits of resistance were low and high-density intervals were large and included zero in many cases. The heritability estimates support the inclusion of resistance to ticks, gastrointestinal nematodes and Eimeria spp. in Nellore breeding programs. Genetic selection can increase the frequency of resistant animals and be used as a complementary tool in parasite control programs. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Calcium Montmorillonite-based dietary supplement attenuates Necrotic Enteritis induced by Eimeria maxima and Clostridium perfringens in broilers

    Science.gov (United States)

    We provide the first description of Dietary Supplement of sorbent minerals attenuates Necrotic Enteritis Induced by Eimeria maxima and Clostridium perfringens in Broilers. Necrotic enteritis (NE) is a poultry disease caused by Clostridium perfringens and characterized by severe intestinal necrosis....

  20. Identification of Eimeria acervulina conoid antigen using chicken monoclonal antibody and liquid chromatography coupled to mass spectrometry

    Science.gov (United States)

    Protozoan parasites of the phylum Apicomplexa include a large number of medically important species. Among them, Toxoplasma gondii, Plasmodium, Cryptosporidium that cause watery diarrhea and mortality in humans and livestock, and Eimeria which induces gastrointestinal disorder in livestock and poul...

  1. Identification and analysis of Eimeria nieschulzi gametocyte genes reveal splicing events of gam genes and conserved motifs in the wall-forming proteins within the genus Eimeria (Coccidia, Apicomplexa

    Directory of Open Access Journals (Sweden)

    Wiedmer Stefanie

    2017-01-01

    Full Text Available The genus Eimeria (Apicomplexa, Coccidia provides a wide range of different species with different hosts to study common and variable features within the genus and its species. A common characteristic of all known Eimeria species is the oocyst, the infectious stage where its life cycle starts and ends. In our study, we utilized Eimeria nieschulzi as a model organism. This rat-specific parasite has complex oocyst morphology and can be transfected and even cultivated in vitro up to the oocyst stage. We wanted to elucidate how the known oocyst wall-forming proteins are preserved in this rodent Eimeria species compared to other Eimeria. In newly obtained genomics data, we were able to identify different gametocyte genes that are orthologous to already known gam genes involved in the oocyst wall formation of avian Eimeria species. These genes appeared putatively as single exon genes, but cDNA analysis showed alternative splicing events in the transcripts. The analysis of the translated sequence revealed different conserved motifs but also dissimilar regions in GAM proteins, as well as polymorphic regions. The occurrence of an underrepresented gam56 gene version suggests the existence of a second distinct E. nieschulzi genotype within the E. nieschulzi Landers isolate that we maintain.

  2. Identification and analysis of Eimeria nieschulzi gametocyte genes reveal splicing events of gam genes and conserved motifs in the wall-forming proteins within the genus Eimeria (Coccidia, Apicomplexa)

    Science.gov (United States)

    Wiedmer, Stefanie; Erdbeer, Alexander; Volke, Beate; Randel, Stephanie; Kapplusch, Franz; Hanig, Sacha; Kurth, Michael

    2017-01-01

    The genus Eimeria (Apicomplexa, Coccidia) provides a wide range of different species with different hosts to study common and variable features within the genus and its species. A common characteristic of all known Eimeria species is the oocyst, the infectious stage where its life cycle starts and ends. In our study, we utilized Eimeria nieschulzi as a model organism. This rat-specific parasite has complex oocyst morphology and can be transfected and even cultivated in vitro up to the oocyst stage. We wanted to elucidate how the known oocyst wall-forming proteins are preserved in this rodent Eimeria species compared to other Eimeria. In newly obtained genomics data, we were able to identify different gametocyte genes that are orthologous to already known gam genes involved in the oocyst wall formation of avian Eimeria species. These genes appeared putatively as single exon genes, but cDNA analysis showed alternative splicing events in the transcripts. The analysis of the translated sequence revealed different conserved motifs but also dissimilar regions in GAM proteins, as well as polymorphic regions. The occurrence of an underrepresented gam56 gene version suggests the existence of a second distinct E. nieschulzi genotype within the E. nieschulzi Landers isolate that we maintain. PMID:29210668

  3. Three operational taxonomic units of Eimeria are common in Nigerian chickens and may undermine effective molecular diagnosis of coccidiosis.

    Science.gov (United States)

    Jatau, Isa D; Lawal, Idris A; Kwaga, Jacob K P; Tomley, Fiona M; Blake, Damer P; Nok, Andrew J

    2016-06-04

    Chicken is fast becoming the world's most consumed meat. As a consequence poultry health is more important now than ever before, with pathogens of chickens recognised as serious threats to food security. One such threat are Eimeria species parasites, protozoa which can cause the disease coccidiosis. Eimeria can compromise economic poultry production and chicken welfare, and have serious consequences for poor livestock keepers. Seven Eimeria species that infect chickens are recognised with a global enzootic distribution. More recently three cryptic Operational Taxonomic Units (OTUx, y and z) have been described in populations of Eimeria recovered from chickens in Australia. Two of the three OTUs have also been detected in sub-Saharan Africa, but their occurrence, pathology and the risk they pose is largely unknown. Nigeria has witnessed a dramatic expansion in poultry production and is now the largest poultry producer in Africa. Here, faecal samples collected from nine of 12 commercial chicken farms sampled in Kaduna state, Nigeria, were found to contain eimerian oocysts. After amplification by in vivo propagation all three cryptic OTU genotypes were detected using polymerase chain reaction (PCR), including OTUy for the first time outside of Australia. Comparison with a widely used, established Eimeria species-specific PCR assay revealed failure to detect the OTU genotypes. All three of the Eimeria OTU genotypes appear to be common in north-western Nigeria. The failure of a leading species-specific molecular assay to detect these genotypes indicates a risk of false negative Eimeria diagnosis when using molecular tools and suggests that the spatial occurrence of each OTU may be far wider than has been recognised. The risk posed by these novel genotypes is unknown, but it is clear that a better understanding of Eimeria occurrence is required together with the validation of effective diagnostics.

  4. Chicken IgY Fc expressed by Eimeria mitis enhances the immunogenicity of E. mitis.

    Science.gov (United States)

    Qin, Mei; Tang, Xinming; Yin, Guangwen; Liu, Xianyong; Suo, Jingxia; Tao, Geru; Ei-Ashram, Saeed; Li, Yuan; Suo, Xun

    2016-03-21

    Eimeria species are obligate intracellular apicomplexan parasites, causing great economic losses in the poultry industry. Currently wild-and attenuated- type anticoccidial vaccines are used to control coccidiosis. However, their use in fast growing broilers is limited by vaccination side effects caused by medium and/or low immunogenic Eimeria spp. There is, therefore, a need for a vaccine with high immunogenicity for broilers. The avian yolk sac IgY Fc is the avian counterpart of the mammalian IgG Fc, which enhances immunogenicity of Fc-fusion proteins. Here, we developed a stable transgenic Eimeria mitis expressing IgY Fc (Emi.chFc) and investigated whether the avian IgY Fc fragment enhances the immunogenicity of E. mitis. Two-week-old broilers were immunized with either Emi.chFc or wild type Eimeria and challenged with wild type E. mitis to analyze the protective properties of transgenic Emi.chFc. Chickens immunized with Emi.chFc had significantly lower oocyst output, in comparison with PBS, mock control (transgenic E. mitis expressing HA1 from H9N2 avian influenza virus) and wildtype E. mitis immunized groups after challenge, indicating that IgY Fc enhanced the immunogenicity of E. mitis. Our findings suggest that IgY Fc-expressing Eimeria may be a better coccidiosis vaccine, and transgenic Eimeria expressing Fc-fused exogenous antigens may be used as a novel vaccine-delivery vehicle against a wide variety of pathogens.

  5. Assessment of the relative success of sporozoite inoculations in individuals exposed to moderate seasonal transmission

    Directory of Open Access Journals (Sweden)

    Spiegel André

    2009-07-01

    Full Text Available Abstract Background The time necessary for malaria parasite to re-appear in the blood following treatment (re-infection time is an indirect method for evaluating the immune defences operating against pre-erythrocytic and early erythrocytic malaria stages. Few longitudinal data are available in populations in whom malaria transmission level had also been measured. Methods One hundred and ten individuals from the village of Ndiop (Senegal, aged between one and 72 years, were cured of malaria by quinine (25 mg/day oral Quinimax™ in three equal daily doses, for seven days. Thereafter, thick blood films were examined to detect the reappearance of Plasmodium falciparum every week, for 11 weeks after treatment. Malaria transmission was simultaneously measured weekly by night collection of biting mosquitoes. Results Malaria transmission was on average 15.3 infective bites per person during the 77 days follow up. The median reappearance time for the whole study population was 46.8 days, whereas individuals would have received an average one infective bite every 5 days. At the end of the follow-up, after 77 days, 103 of the 110 individuals (93.6%; CI 95% [89.0–98.2] had been re-infected with P. falciparum. The median reappearance time ('re-positivation' was longer in subjects with patent parasitaemia at enrolment than in parasitologically-negative individuals (58 days vs. 45.9; p = 0.03 and in adults > 30 years than in younger subjects (58.6 days vs. 42.7; p = 0.0002. In a multivariate Cox PH model controlling for the sickle cell trait, G6PD deficiency and the type of habitat, the presence of parasitaemia at enrolment and age ≥ 30 years were independently predictive of a reduced risk of re-infection (PH = 0.5 [95% CI: 0.3–0.9] and 0.4; [95% CI: 0.2–0.6] respectively. Conclusion Results indicate the existence of a substantial resistance to sporozoites inoculations, but which was ultimately overcome in almost every individual after 2 1/2 months of

  6. Nuevos casos de coccidiosis bovina en León. Denuncia de Eimeria bovis (Züblin,1908) Fiebiger, 1912, E. auburnensis Christensen y Porter, 1939 y E. ellipsoidalis Becker y Frye, 1929

    OpenAIRE

    Cordero del Campillo, Miguel

    1981-01-01

    P. 61-72 Se estudian nuevos focos de coccidiosis bovina en la provincia de León. En la zona montañosa del NE, se identifica, nuevamente Eimeria zürni más Eimeria auburnensis. En las cercanías de la ciudad de León se comprobó la existencia de Eimeria bovis, Eimeria auburnensis y Eimeria ellipsoidalis. La máxima frecuencia corre a cargo de Eimeria zürni y Eimeria bovis, dotadas también de mayor poder patógeno. Los datos morfológicos de Eimeria zürni concuerdan con los estudiados por el autor...

  7. Analysis of intrinsic and extrinsic factors influencing the dynamics of bovine Eimeria spp. from central-eastern Poland.

    Science.gov (United States)

    Tomczuk, Krzysztof; Grzybek, Maciej; Szczepaniak, Klaudiusz; Studzińska, Maria; Demkowska-Kutrzepa, Marta; Roczeń-Karczmarz, Monika; Klockiewicz, Maciej

    2015-11-30

    Eimeria infections are common in cattle worldwide, however, little is known about the invasion dynamics of this unicellular parasite. Therefore, the aim of this study was to analyze intrinsic (host age) and extrinsic (herd size and management system) factors influencing the dynamics of Eimeria spp. found in calves from CE Poland. Fecal samples were collected from 356 calves from different types of management systems and from different herd sizes. Flotation and McMaster method were used for parasitological investigation. Oocysts were differentiated on the basis of morphological criteria. Eight Eimeria species were identified and mean species richness (MSR) was significantly affected by host age. The highest MSR was noted for middle age animals. There was an association between species, with a highly significant co-occurrence of Eimeria bovis with Eimeria zuernii. The presence of E. bovis significantly increased the percentage of individuals carrying E. zuernii. The presence of E. bovis significantly increased the percentage of individuals carrying Eimeria canadensis. The overall prevalence of Eimeria spp. reached 52.8% and was significantly affected by the age of cows, with the highest prevalence in animals between 5-10 months old. The most prevalent species were E. bovis (37.4%), E. zuernii (19.9%) and E. canadensis (12.1%). The prevalence of E. bovis was affected by host age (the highest prevalence in age class 2 animals) and management type (the highest prevalence in individuals raised in groups). The prevalence of E. zuernii was affected by age (the lowest prevalence was noted in the oldest individuals) and herd size (individuals infected were present only in the middle and large size herds), whereas the prevalence of E. canadensis was affected by all three factors. Overall, mean OPG of the combined Eimeria spp. was 458.84 (37.93) and differed significantly between age classes. Mean OPGs were generally low for young and mature animals but high for middle age

  8. Oral antibody to interleukin-10 reduces growth rate depression due to Eimeria spp. infection in broiler chickens.

    Science.gov (United States)

    Sand, Jordan M; Arendt, Maria K; Repasy, Alec; Deniz, Gűlay; Cook, Mark E

    2016-02-01

    Eimeria spp. must be controlled in floor-reared poultry to prevent the onset of coccidiosis. Here we use an oral antibody to chicken IL-10 to prevent growth depression due to Eimeria spp. infection. Egg antibody directed against an antigenic peptide of IL-10 was produced in laying hens and measured using an ELISA. In the first experiment, egg yolk powder containing antibody to chicken IL-10 (vlpramqt conjugate) (anti-IL-10 yolk powder) was fed at 3.4 g/kg feed to determine growth response following mixed Eimeria spp. challenge. Chicks were fed either anti-IL-10 antibodies or control antibodies and challenged (d3) with either sterile saline or a 10× attenuated Eimeria spp. vaccine. Control-fed and Eimeria-challenged chicks grew 8.8% slower than those challenged with saline (P < 0.04), whereas anti-IL-10-fed Eimeria challenged chicks were not different from untreated controls. In the second trial a dose response was performed with doses of either 0 (control antibody), 0.34-, or 3.4-g anti-IL-10 yolk powder/kg feed. Control-fed, Eimeria-challenged chicks grew 10.6% slower than control saline-challenged chicks (P < 0.05); however, anti-IL-10-fed chicks fed either dose of anti-IL-10 were not different from saline-challenged chicks. Finally, the effect of anti-IL-10 on acquired immunity was investigated. Chicks were fed control or anti-IL-10 yolk powder and vaccinated with a 1× dose of Eimeria vaccine at d 3. After 14 d, antibody was removed from the diet. Chicks were either saline or 10× Eimeria challenged at d 17. We found that the anti-IL-10-fed chickens did not show a reduction in growth due to challenge; hence anti-IL-10 does not appear to affect adaptive immunity during the primary immunization. Overall, use of an antibody to IL-10 is a novel method in preventing adverse effects of Eimeria spp. infection in poultry. © 2016 Poultry Science Association Inc.

  9. An assessment in rodents of the pathological and immunopathological consequence of multiple vaccinations and challenge with radiation-attenuated malaria parasites (blood forms and sporozoites)

    International Nuclear Information System (INIS)

    Boonpucknavig, S.

    1982-06-01

    Multiple vaccination with irradiated merozoites of Plasmodium berghei resulted in high levels of circulating antibody but a low degree of protection to challenge with normal merozoites. On the other hand, the multiple vaccinations and the challenge resulted in severe immunopathological reactions shortly after the immunization or challenge. These reactions were seen in the liver, kidneys and spleen and included the accumulation of mononuclear cells, severe coagulative necrosis of liver cells and proliferative changes in the splenic white pulp and in the glomeruli. The pathological reactions were more severe than in non-immunized animals but the parasitemia was lower and less malarial antigen was detected in Kupfer Cells of the liver, the sinusoidal cells of the spleen and the reticuloendothelial cells in the interstitial tissue of the kidney. Vaccination with irradiated sporozoites of P. berghei resulted in good protection to challenge with normal sporozoites even before circulating anti-sporozoite antibody could be detected. Only mild pathological changes were associated with up to 4 immunizations followed by challenge and these were largely limited to the liver, and were reversible. Sporozoite antigens were detected in the spleen and immune complexes in the glomeruli for 2-4 weeks following challenge but not later. Immunized mice however often developed some lobular pneumonia of the lung but the severity of this did not increase with challenge

  10. Long term protection after immunization with P. berghei sporozoites correlates with sustained IFNγ responses of hepatic CD8+ memory T cells.

    Science.gov (United States)

    Nganou-Makamdop, Krystelle; van Gemert, Geert-Jan; Arens, Theo; Hermsen, Cornelus C; Sauerwein, Robert W

    2012-01-01

    Protection against P. berghei malaria can successfully be induced in mice by immunization with both radiation attenuated sporozoites (RAS) arresting early during liver stage development, or sporozoites combined with chloroquine chemoprophylaxis (CPS), resulting in complete intra-hepatic parasite development before killing of blood-stages by chloroquine takes place. We assessed the longevity of protective cellular immune responses by RAS and CPS P. berghei immunization of C57BL/6j mice. Strong effector and memory (T(EM)) CD8+ T cell responses were induced predominantly in the liver of both RAS and CPS immunized mice while CD4+ T cells with memory phenotype remained at base line levels. Compared to unprotected naïve mice, we found high sporozoite-specific IFNγ ex vivo responses that associated with induced levels of in vivo CD8+ T(EM) cells in the liver but not spleen. Long term evaluation over a period of 9 months showed a decline of malaria-specific IFNγ responses in RAS and CPS mice that significantly correlated with loss of protection (r(2) = 0.60, pmemory CD8+ T cells could be boosted by re-exposure to wild-type sporozoites. Our data show that sustainable protection against malaria associates with distinct intra-hepatic immune responses characterized by strong IFNγ producing CD8+ memory T cells.

  11. Long term protection after immunization with P. berghei sporozoites correlates with sustained IFNγ responses of hepatic CD8+ memory T cells.

    Directory of Open Access Journals (Sweden)

    Krystelle Nganou-Makamdop

    Full Text Available Protection against P. berghei malaria can successfully be induced in mice by immunization with both radiation attenuated sporozoites (RAS arresting early during liver stage development, or sporozoites combined with chloroquine chemoprophylaxis (CPS, resulting in complete intra-hepatic parasite development before killing of blood-stages by chloroquine takes place. We assessed the longevity of protective cellular immune responses by RAS and CPS P. berghei immunization of C57BL/6j mice. Strong effector and memory (T(EM CD8+ T cell responses were induced predominantly in the liver of both RAS and CPS immunized mice while CD4+ T cells with memory phenotype remained at base line levels. Compared to unprotected naïve mice, we found high sporozoite-specific IFNγ ex vivo responses that associated with induced levels of in vivo CD8+ T(EM cells in the liver but not spleen. Long term evaluation over a period of 9 months showed a decline of malaria-specific IFNγ responses in RAS and CPS mice that significantly correlated with loss of protection (r(2 = 0.60, p<0.0001. The reducing IFNγ response by hepatic memory CD8+ T cells could be boosted by re-exposure to wild-type sporozoites. Our data show that sustainable protection against malaria associates with distinct intra-hepatic immune responses characterized by strong IFNγ producing CD8+ memory T cells.

  12. Plasmodium berghei sporozoite challenge of vaccinated BALB/c mice leads to the induction of humoral immunity and improved function of CD8(+) memory T cells

    Czech Academy of Sciences Publication Activity Database

    Tartz, S.; Deschermeier, Ch.; Retzlaff, S.; Heussler, V.; Šebo, Peter; Fleischer, B.; Jacobs, T.

    2013-01-01

    Roč. 43, č. 3 (2013), s. 693-704 ISSN 0014-2980 R&D Projects: GA ČR GA310/08/0447 Institutional research plan: CEZ:AV0Z50200510 Keywords : CD8+Tcells * Plasmodium * Sporozoites Subject RIV: EE - Microbiology, Virology Impact factor: 4.518, year: 2013

  13. The impact of livestock on the abundance, resting behaviour and sporozoite rate of malaria vectors in southern Tanzania.

    Science.gov (United States)

    Mayagaya, Valeriana S; Nkwengulila, Gamba; Lyimo, Issa N; Kihonda, Japheti; Mtambala, Hassan; Ngonyani, Hassan; Russell, Tanya L; Ferguson, Heather M

    2015-01-21

    Increases in the coverage of long-lasting insecticidal nets (LLINs) have significantly reduced the abundance of Anopheles gambiae sensu stricto in several African settings, leaving its more zoophagic sibling species Anopheles arabiensis as the primary vector. This study investigated the impact of livestock ownership at the household level on the ecology and malaria infection rate of vectors in an area of Tanzania where An. arabiensis accounts for most malaria transmission. Mosquito vectors were collected resting inside houses, animal sheds and in outdoor resting boxes at households with and without livestock over three years in ten villages of the Kilombero Valley, Tanzania. Additionally, the abundance and sporozoite rate of vectors attempting to bite indoors at these households was assessed as an index of malaria exposure. The mean abundance of An. gambiae s.l. biting indoors was similar at houses with and without livestock. In all years but one, the relative proportion of An. arabiensis within the An. gambiae s.l. species complex was higher at households with livestock. Livestock presence had a significant impact on malaria vector feeding and resting behaviour. Anopheles arabiensis were generally found resting in cattle sheds where livestock were present, and inside houses when absent. Correspondingly, the human blood index of An. arabiensis and An. funestus s.l. was significant reduced at households with livestock, whereas that of An. gambiae s.s. was unaffected. Whilst there was some evidence that sporozoite rates within the indoor-biting An. gambiae s.l population was significantly reduced at households with livestock, the significance of this effect varied depending on how background spatial variation was accounted for. These results confirm that the presence of cattle at the household level can significantly alter the local species composition, feeding and resting behaviour of malaria vectors. However, the net impact of this livestock-associated variation in

  14. Seroprevalence of Antibodies against Plasmodium falciparum Sporozoite Antigens as Predictive Disease Transmission Markers in an Area of Ghana with Seasonal Malaria Transmission.

    Directory of Open Access Journals (Sweden)

    Kwadwo A Kusi

    Full Text Available As an increasing number of malaria-endemic countries approach the disease elimination phase, sustenance of control efforts and effective monitoring are necessary to ensure success. Mathematical models that estimate anti-parasite antibody seroconversion rates are gaining relevance as more sensitive transmission intensity estimation tools. Models however estimate yearly seroconversion and seroreversion rates and usually predict long term changes in transmission, occurring years before the time of sampling. Another challenge is the identification of appropriate antigen targets since specific antibody levels must directly reflect changes in transmission patterns. We therefore investigated the potential of antibodies to sporozoite and blood stage antigens for detecting short term differences in malaria transmission in two communities in Northern Ghana with marked, seasonal transmission.Cross-sectional surveys were conducted during the rainy and dry seasons in two communities, one in close proximity to an irrigation dam and the other at least 20 Km away from the dam. Antibodies against the sporozoite-specific antigens circumsporozoite protein (CSP and Cell traversal for ookinetes and sporozoites (CelTOS and the classical blood stage antigen apical membrane antigen 1 (AMA1 were measured by indirect ELISA. Antibody levels and seroprevalence were compared between surveys and between study communities. Antibody seroprevalence data were fitted to a modified reversible catalytic model to estimate the seroconversion and seroreversion rates.Changes in sporozoite-specific antibody levels and seroprevalence directly reflected differences in parasite prevalence between the rainy and dry seasons and hence the extent of malaria transmission. Seroconversion rate estimates from modelled seroprevalence data did not however support the above observation.The data confirms the potential utility of sporozoite-specific antigens as useful markers for monitoring short term

  15. Eimeria Species in Danish Dairy Cattle – Preliminary Data from an Ongoing Study

    DEFF Research Database (Denmark)

    Enemark, Heidi L.; Enemark, J. M.D.

    2011-01-01

    . auburnensis was in several cases correlated to diarrhea. These cases however were not diagnosed as coccidiosis. The results warrants further pathogenicity studies of the different Eimeria spp. In addition, it was shown that correct diagnosis of coccidiosis is a challenge and knowledge of the management system......Contrary to the majority of European countries, antiparasiticides are on prescription only in Denmark, thus treatment requires a proper diagnosis made by a veterinarian, and therefore relies on adequate diagnostic procedures. This study was performed to obtain information about presence of Eimeria...... identified so far. Of the faecal samples included in the study 7% had a firm/ normal consistency, 81% were soft to liquid, and 12 % were watery with blood and/or mucus. Oocyst excretion above 5000 oocysts per gram (OPG) was found in 6.5% of the calves, whereas 12.0% excreted 500-5000 OPG. Clinical...

  16. Outbreak of Eimeria kofoidi and E. legionensis coccidiosis in redlegged partridges (Alectoris rufa

    Directory of Open Access Journals (Sweden)

    Pier Giorgio Bolognesi

    2006-01-01

    Full Text Available An outbreak of coccidiosis occurred in red-legged partridges is reported. At the post-mortem examination the birds showed a mucous haemorrhagic enteritis, mostly in the duodenal intestinal tract. Direct microscopic examination of intestinal content revealed the presence of a high number of oocysts. After incubation, on the basis of the morphological features, two species of coccidia were identified: Eimeria kofoidi and E. legionensis.

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

    Science.gov (United States)

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

    2018-02-15

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

  18. Two new species of Eimeria from peacocks (Pavo cristatus) in Saudi Arabia.

    Science.gov (United States)

    Amoudi, M A

    1988-11-01

    Fifteen fecal samples from peacocks (Pavo cristatus) in Saudi Arabia contained oocysts of Eimeria riyadhae n. sp. in two peacocks and oocysts of E. arabica n. sp. in one peacock. Sporulated oocysts of Eimeria riyadhae are ellipsoidal, 27-30.5 x 20.5-25 (28.8 +/- 1.3 x 22.4 +/- 1.6) micron, with a two-layered wall and bilobed polar body, but without a micropyle or residuum. The sporocysts are ovoid, 11-14.5 x 6.5-8 (13.2 +/- 1.2 x 7.2 +/- 0.6) micron with a thick, knob-like Stieda body and a residuum. Sporulated oocysts of Eimeria arabica are spheroidal, 17.5-21.5 x 17.5-21.5 (19.2 +/- 1.6 x 19.2 +/- 1.6) micron, with a two-layered wall and two refractile polar bodies, but without a micropyle or residuum. The sporocyts are elongate ovoid, 9.5-12 x 4-6.5 (11.2 +/- 0.9 x 5.5 +/- 0.88), with a small crescent-shaped Stieda body. The host bird belongs to the order Galliformis.

  19. New species of Eimeria (Apicomplexa: Eimeriidae) from Thrichomys fosteri and Clyomys laticeps (Rodentia: Echimyidae) of the Brazilian Pantanal.

    Science.gov (United States)

    Barreto, Wanessa Teixeira Gomes; Viana, Lúcio André; Santos, Filipe Martins; de Oliveira Porfírio, Grasiela Edith; Perdomo, Alessandra Cabral; da Silva, Alanderson Rodrigues; de Sousa, Keyla Carstens Marques; de Oliveira, Michel Angelo Constantino; Herrera, Heitor Miraglia; de Andrade, Gisele Braziliano

    2017-11-01

    The echimyid rodents Thrichomys fosteri and Clyomys laticeps are among the most commonly recorded small mammals in the Pantanal wetland of Brazil. These species play important ecological roles since they are the basis of the food chain of some predators and are parasitized by some pathogens. Knowledge of the eimerians that parasitize echimyid rodents in Brazil is absent, and only one report is available for South America. We therefore investigated parasitism by coccidians in the echimyids T. fosteri and C. laticeps in the Pantanal. Using morphological and morphometric features and associated statistical analyses, we describe five new eimerian species parasitizing T. fosteri (Eimeria nhecolandensis n. sp., Eimeria jansenae n. sp., and Eimeria fosteri n. sp.) and C. laticeps (E. nhecolandensis n. sp., Eimeria corumbaensis n. sp., and Eimeria laticeps n. sp.) in different types of infection associations. We document the developmental forms in the tissues, and describe lesions in the enteric tract of some infected animals. We also discuss some approaches regarding epidemiological and ecological data. Our results demonstrate that echimyid rodents in the Brazilian Pantanal are important hosts for the maintenance of enteric coccidia. Moreover, in some circumstances, this parasitism may threaten the health of the hosts.

  20. Loop-mediated isothermal amplification (LAMP) assays for the species-specific detection of Eimeria that infect chickens.

    Science.gov (United States)

    Barkway, Christopher P; Pocock, Rebecca L; Vrba, Vladimir; Blake, Damer P

    2015-02-20

    Eimeria species parasites, protozoa which cause the enteric disease coccidiosis, pose a serious threat to the production and welfare of chickens. In the absence of effective control clinical coccidiosis can be devastating. Resistance to the chemoprophylactics frequently used to control Eimeria is common and sub-clinical infection is widespread, influencing feed conversion ratios and susceptibility to other pathogens such as Clostridium perfringens. Despite the availability of polymerase chain reaction (PCR)-based tools, diagnosis of Eimeria infection still relies almost entirely on traditional approaches such as lesion scoring and oocyst morphology, but neither is straightforward. Limitations of the existing molecular tools include the requirement for specialist equipment and difficulties accessing DNA as template. In response a simple field DNA preparation protocol and a panel of species-specific loop-mediated isothermal amplification (LAMP) assays have been developed for the seven Eimeria recognised to infect the chicken. We now provide a detailed protocol describing the preparation of genomic DNA from intestinal tissue collected post-mortem, followed by setup and readout of the LAMP assays. Eimeria species-specific LAMP can be used to monitor parasite occurrence, assessing the efficacy of a farm's anticoccidial strategy, and to diagnose sub-clinical infection or clinical disease with particular value when expert surveillance is unavailable.

  1. Mitochondrial genomes of Australian chicken Eimeria support the presence of ten species with low genetic diversity among strains.

    Science.gov (United States)

    Morgan, Jess A T; Godwin, Rosamond M

    2017-08-30

    Modern molecular approaches have vastly improved diagnostic capabilities for differentiating among species of chicken infecting Eimeria. Consolidating information from multiple genetic markers, adding additional poultry Eimeria species and increasing the size of available data-sets is improving the resolving power of the DNA, and consequently our understanding of the genus. This study adds information from 25 complete mitochondrial DNA genomes from Australian chicken Eimeria isolates representing all 10 species known to occur in Australia, including OTU-X, -Y and -Z. The resulting phylogeny provides a comprehensive view of species relatedness highlighting where the OTUs align with respect to others members of the genus. All three OTUs fall within the Eimeria clade that contains only chicken-infecting species with close affinities to E. maxima, E. brunetti and E. mitis. Mitochondrial genetic diversity was low among Australian isolates likely reflecting their recent introduction to the country post-European settlement. The lack of observed genetic diversity is a promising outcome as it suggests that the currently used live vaccines should continue to offer widespread protection against Eimeria outbreaks in all states and territories. Flocks were frequently found to host multiple strains of the same species, a factor that should be considered when studying disease epidemiology in the field. Crown Copyright © 2017. Published by Elsevier B.V. All rights reserved.

  2. Studies on the in vitro cultivation of coccidia

    International Nuclear Information System (INIS)

    Schmatz, D.M.

    1985-01-01

    New approaches to the in vitro cultivation of coccidian parasites are described here, specifically for avian coccidia of the genus Eimeria. Firstly, an improved method of purifying the infectious stage of these parasites, known as sporozoites, over a DEAE-52 cellulose anion exchange column to eliminate toxic debris generated during excystation is described. The cultured cells used to support the intracellular development of these parasites, Madin-Darby Bovine Kidney Cells (MDBK), were cloned and it was demonstrated that some clones were more susceptible than others to infection with sporozoites. The use of sub-lethal doses of gamma radiation to pre-treat host cell monolayers prior to infecting has been found to prevent host cell overgrowth and subsequent peeling of the monolayers while not interfering with parasite development. Utilizing in vitro culture techniques developed here in conjunction with radiolabeling studies, an assay has been development using the parasite-specific incorporation of 3 H-uracil to assess the intracellular development of E. tenella and E. acervulina in vitro. As shown by both scintillation counts and autoradiography, 3 H-uracil was incorporated specifically into the intracellular parasites from the onset of infection and continued throughout the development of the first generation schizonts. Based on these findings, a semi-automated microscale incorporation assay was developed to determine parasite viability. The assay system is used in this study to investigate the effects of known anticoccidials, sporozoite antiserum, and varying the composition of the cell culture medium on parasite development

  3. Pathogenic characteristics of a novel intranuclear coccidia in Japanese black calves and its genetic identification as Eimeria subspherica.

    Science.gov (United States)

    Koreeda, Terunori; Kawakami, Tomo; Okada, Ayako; Hirashima, Yoshimasa; Imai, Naoto; Sasai, Kazumi; Tanaka, Shogo; Matsubayashi, Makoto; Shibahara, Tomoyuki

    2017-11-01

    Bovine intranuclear coccidiosis is caused by the protozoans Eimeria alabamensis and Cyclospora spp. Here, we characterized the disease and genetically identified the causative species in Japanese black calves with chronic and refractory watery diarrhea. Histologic examinations revealed atrophy of the jejunal villi and numerous parasites in the nucleus of epithelial cells in the jejunum. Based on molecular analyses using 18S ribosomal RNA gene-specific primers that we designed, the parasites were found to be formed in the same cluster as Eimeria subspherica in the phylogenetic tree, which was separated from those of other related Eimeria spp. These results constitute the first report of E. subspherica as a cause of bovine intranuclear coccidiosis.

  4. Two necrotic enteritis predisposing factors, dietary fishmeal and Eimeria infection, induce large changes in the caecal microbiota of broiler chickens.

    Science.gov (United States)

    Wu, Shu-Biao; Stanley, Dragana; Rodgers, Nicholas; Swick, Robert A; Moore, Robert J

    2014-03-14

    It is widely established that a high-protein fishmeal supplemented starter diet and Eimeria infection can predispose birds to the development of clinical necrotic enteritis symptoms following Clostridium perfringens infection. However, it has not been clearly established what changes these treatments cause to predispose birds to succumb to necrotic enteritis. We analysed caecal microbiota of 4 groups of broilers (n=12) using deep pyrosequencing of 16S rDNA amplicons: (1) control chicks fed a control diet, (2) Eimeria infected chicks fed control diet, (3) chicks fed fishmeal supplemented diet and lastly (4) both fishmeal fed and Eimeria infected chicks. We found that the high-protein fishmeal diet had a strong effect on the intestinal microbiota similar to the previously reported effect of C. perfringens infection. We noted major changes in the prevalence of various lactobacilli while the total culturable Lactobacillus counts remained stable. The Ruminococcaceae, Lachnospiraceae, unknown Clostridiales and Lactobacillaceae families were most affected by fishmeal with increases in a number of operational taxonomic units (OTUs) that had previously been linked to Crohn's disease and reductions in OTUs known to be butyrate producers. Eimeria induced very different changes in microbiota; Ruminococcaceae groups were reduced in number and three unknown Clostridium species were increased in abundance. Additionally, Eimeria did not significantly influence changes in pH, formic, propionic or isobutyric acid while fishmeal induced dramatic changes in all these measures. Both fishmeal feeding and Eimeria infection induced significant changes in the gut microbiota; these changes may play an important role in predisposing birds to necrotic enteritis. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Eimeria pavoaegyptica sp. nov. (Apicomplexa: Eimeriidae) in faeces of Indian peacocks, Pavo cristatus Linnaeus, 1758 (Galliformes: Phasianidae) from Egypt

    OpenAIRE

    El-Shahawy,Ismail Saad

    2010-01-01

    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 ooc...

  6. Incidence of single and mixed infections with Eimeria kofoidi, E. caucasica and E. legionensis on the health of experimentally infected red-legged partridges (Alectoris rufa).

    Science.gov (United States)

    Naciri, M; Fort, G; Briant, J; Duperray, J; Benzoni, G

    2014-09-15

    Little is known about Eimeria-induced coccidiosis in partridges. After a coccidiosis outbreak in a farm rearing red-legged partridges (Alectoris rufa) in Brittany (France), three Eimeria species were identified as Eimeria kofoidi, Eimeria caucasica and Eimeria legionensis. This study aimed to reproduce the effects of the disease occurring in field conditions, in the absence of preventive treatments, to further build a coccidiosis model, helpful for coccidiostatic development. The pathogenic effects of a single infection with Eimeria kofoidi, E. caucasica and E. legionensis were evaluated, as well as the effects of multiple infections associating two or three of these species in red-legged partridges. Thirty-one-day-old birds were individually inoculated with Eimeria spp. and clinically followed up until 49 days of age. Mortality, lesion scores, daily oocyst production and growth were used as assessment criteria. Single infections with 250,000 E. kofoidi, 30,000 E. caucasica or 100,000 E. legionensis oocysts did not increase mortality rate compared to uninfected birds, whereas the combination of 3 species caused significant 28% mortality (PEimeria spp. or for selecting efficient molecules to struggle coccidiosis of red-legged partridges. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. The development of exo-erythrocytic schizonts of Plasmodium berghei in vitro from gamma-irradiated and non-irradiated sporozoites: a study using confocal laser scanning microscopy

    International Nuclear Information System (INIS)

    Sinden, E.; Couchman, A.; Suhrbier, A.; Marsh, F.; Winger, L.; Ranawaka, G.

    1991-01-01

    Confocal scanning laser microscopy has been used to study the distribution of antigens expressed by the liver stages of Plasmodium berghei in cultured hepatoma cells. The 3-dimensional images obtained of intact parasites clearly show complex patterns of antigen expression not apparent when using conventional IFAT or immunoelectron microscopy. A liver-stage specific antigen (Pbl 1) was shown to be confined to the parasitophorous vacuole; the vacuole has extensive diverticulae extending into the host cell. Small parasites were detected for the first time in 'mature' cultures. These did not represent a distinct population, but the 'tail' of a broad continuum of parasite sizes. Irradiated sporozoites produce a transient population of slow-growing parasites which express a very limited range of antigens de novo in the invaded hepatoma cell. A comparison of the reactivity of normal EE parasites with anti-circumsporozoite antibody and with ant-Pbl 1 suggests that the former reagent may reliably be used to identify sporozoites invading host cells, but should not be used to determine the number of parasites that successfully undergo intrahepatic development. Anti-Pbl-1 indicates on 33% of invaded sporozoites identified by anti-CSP subsequently differentiate. (author)

  8. Combination of probiotics and coccidiosis vaccine enhances protection against an Eimeria challenge.

    Science.gov (United States)

    Ritzi, Miranda M; Abdelrahman, Wael; van-Heerden, Kobus; Mohnl, Michaela; Barrett, Nathaniel W; Dalloul, Rami A

    2016-11-08

    Coccidiosis is endemic in the commercial broiler industry capable of inflicting devastating economic losses to poultry operations. Vaccines are relatively effective in controlling the disease; their efficacy could potentially be improved with concurrent use of probiotics as evaluated in this study using an Eimeria challenge. Day of hatch 400 Cobb-500 male broilers were assigned to one of four treatment groups including control (CON), vaccine-only gel application (VNC), probiotic-only gel application (NPC), and vaccine-plus-probiotic gel application (VPC). Birds were placed in floor pens (6 replicate pens/treatment, 16-17 birds/pen). NPC and VPC birds received the probiotics in the water on days 2-4, 8, 14-20, 22, 29, and 34-36. On day 15, birds were mildly challenged with 0.5 mL of a mixed oral inoculum of Eimeria sp. prepared with the coccidiosis vaccine at 10× the vaccination dose. Performance measurements were recorded on first day and weekly afterwards, and lesion scores were evaluated 6 days post-challenge. Overall, the probiotics and coccidiosis vaccine resulted in an enhanced protective effect against the challenge, with VPC birds exhibiting lower lesion scores in the duodenum than VNC or NPC birds. Birds in the VPC treatment also demonstrated higher weight gains during days 1-15, days 7-15, and days 21-28 when compared to the VNC birds. These results suggest that the combination of probiotics and coccidiosis vaccines could enhance performance and provide an additional protective effect against a mixed Eimeria challenge.

  9. Age-related immune response to experimental infection with Eimeria ninakohlyakimovae in goat kids.

    Science.gov (United States)

    Matos, L; Muñoz, M C; Molina, J M; Rodríguez, F; Pérez, D; López, A M; Hermosilla, C; Taubert, A; Ruiz, A

    2018-06-01

    Both the immune response developed in ruminants against Eimeria spp. and the ability to bear patent infections seems to be dependent on the age of the host. In the present study we have evaluated the influence of the age in the development of protective immune responses against Eimeria ninakohlyakimovae. For this purpose, 3, 4 and 5-week-old goat kids were infected with sporulated oocysts and subjected to a homologous challenge 3 weeks later. Goat kids primary infected at 6, 7 and 8 weeks of age served as challenge controls, and uninfected animals were used as negative controls. The protective immunity was assessed by clinical, haematological, parasitological, immunological and pathological parameters. Altogether, the results demonstrate that goat kids of either 3, 4 or 5 weeks of age are able to develop patent infections and immunoprotective responses against E. ninakohlyakimovae, as all age groups: (i) released significantly less oocysts after challenge, which was associated to milder clinical signs; (ii) displayed a local immune response, with significant increase of numerous cellular populations; and (iii) had increased levels of IgG and IgM, and mainly of local IgA. Nevertheless, detailed analysis of the data showed some differences between the three age groups, related both to the Eimeria infection outcome and the resulting immune response, suggesting that youngest goat kids are not fully immunocompetent. This finding may be of interest for the design of immunoprophylactic approaches and/or prophylactic/methaphylactic treatments against goat coccidiosis. Copyright © 2018 Elsevier Ltd. All rights reserved.

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

    International Nuclear Information System (INIS)

    Penev, P.; Stefanova, M.

    1975-01-01

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

  11. Sporozoite Route of Infection Influences In Vitro var Gene Transcription of Plasmodium falciparum Parasites From Controlled Human Infections.

    Science.gov (United States)

    Dimonte, Sandra; Bruske, Ellen I; Hass, Johanna; Supan, Christian; Salazar, Carmen L; Held, Jana; Tschan, Serena; Esen, Meral; Flötenmeyer, Matthias; Koch, Iris; Berger, Jürgen; Bachmann, Anna; Sim, Betty K L; Hoffman, Stephen L; Kremsner, Peter G; Mordmüller, Benjamin; Frank, Matthias

    2016-09-15

    Antigenic variation in Plasmodium falciparum is mediated by the multicopy var gene family. Each parasite possesses about 60 var genes, and switching between active var loci results in antigenic variation. In the current study, the effect of mosquito and host passage on in vitro var gene transcription was investigated. Thirty malaria-naive individuals were inoculated by intradermal or intravenous injection with cryopreserved, isogenic NF54 P. falciparum sporozoites (PfSPZ) generated from 1 premosquito culture. Microscopic parasitemia developed in 22 individuals, and 21 in vitro cultures were established. The var gene transcript levels were determined in early and late postpatient cultures and in the premosquito culture. At the early time point, all cultures preferentially transcribed 8 subtelomeric var genes. Intradermal infections had higher var gene transcript levels than intravenous infections and a significantly longer intrahost replication time (P = .03). At the late time point, 9 subtelomeric and 8 central var genes were transcribed at the same levels in almost all cultures. Premosquito and late postpatient cultures transcribed the same subtelomeric and central var genes, except for var2csa  The duration of intrahost replication influences in vitro var gene transcript patterns. Differences between premosquito and postpatient cultures decrease with prolonged in vitro growth. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

  12. Occurrence of Eimeria species parasites on small-scale commercial chicken farms in Africa and indication of economic profitability.

    Directory of Open Access Journals (Sweden)

    Kimberly M Fornace

    Full Text Available Small-scale commercial poultry production is emerging as an important form of livestock production in Africa, providing sources of income and animal protein to many poor households, yet the occurrence and impact of coccidiosis on this relatively new production system remains unknown. The primary objective of this study was to examine Eimeria parasite occurrence on small-scale commercial poultry farms in Ghana, Tanzania and Zambia. Additionally, farm economic viability was measured by calculating the farm gross margin and enterprise budget. Using these economic measures as global assessments of farm productivity, encompassing the diversity present in regional husbandry systems with a measure of fundamental local relevance, we investigated the detection of specific Eimeria species as indicators of farm profitability. Faecal samples and data on production parameters were collected from small-scale (less than 2,000 birds per batch intensive broiler and layer farms in peri-urban Ghana, Tanzania and Zambia. All seven Eimeria species recognised to infect the chicken were detected in each country. Furthermore, two of the three genetic variants (operational taxonomic units identified previously in Australia have been described outside of Australia for the first time. Detection of the most pathogenic Eimeria species associated with decreased farm profitability and may be considered as an indicator of likely farm performance. While a causal link remains to be demonstrated, the presence of highly pathogenic enteric parasites may pose a threat to profitable, sustainable small-scale poultry enterprises in Africa.

  13. Occurrence of Eimeria Species Parasites on Small-Scale Commercial Chicken Farms in Africa and Indication of Economic Profitability

    Science.gov (United States)

    Fornace, Kimberly M.; Clark, Emily L.; Macdonald, Sarah E.; Namangala, Boniface; Karimuribo, Esron; Awuni, Joseph A.; Thieme, Olaf; Blake, Damer P.; Rushton, Jonathan

    2013-01-01

    Small-scale commercial poultry production is emerging as an important form of livestock production in Africa, providing sources of income and animal protein to many poor households, yet the occurrence and impact of coccidiosis on this relatively new production system remains unknown. The primary objective of this study was to examine Eimeria parasite occurrence on small-scale commercial poultry farms in Ghana, Tanzania and Zambia. Additionally, farm economic viability was measured by calculating the farm gross margin and enterprise budget. Using these economic measures as global assessments of farm productivity, encompassing the diversity present in regional husbandry systems with a measure of fundamental local relevance, we investigated the detection of specific Eimeria species as indicators of farm profitability. Faecal samples and data on production parameters were collected from small-scale (less than 2,000 birds per batch) intensive broiler and layer farms in peri-urban Ghana, Tanzania and Zambia. All seven Eimeria species recognised to infect the chicken were detected in each country. Furthermore, two of the three genetic variants (operational taxonomic units) identified previously in Australia have been described outside of Australia for the first time. Detection of the most pathogenic Eimeria species associated with decreased farm profitability and may be considered as an indicator of likely farm performance. While a causal link remains to be demonstrated, the presence of highly pathogenic enteric parasites may pose a threat to profitable, sustainable small-scale poultry enterprises in Africa. PMID:24391923

  14. Volutin granules of Eimeria parasites are acidic compartments and have physiological and structural characteristics similar to acidocalcisomes

    Science.gov (United States)

    Medeiros, Lia Carolina Soares; Gomes, Fabio; Maciel, Luis Renato Maia; Seabra, Sergio Henrique; Docampo, Roberto; Moreno, Silvia; Plattner, Helmut; Hentschel, Joachim; Kawazoe, Urara; Barrabin, Hector; de Souza, Wanderley; DaMatta, Renato Augusto; Miranda, Kildare

    2012-01-01

    The structural organization of parasites has been the subject of investigation by many groups and has lead to the identification of structures and metabolic pathways that may represent targets for anti-parasitic drugs. A specific group of organelles named acidocalcisomes has been identified in a number of organisms, including the apicomplexan parasites such as Toxoplasma and Plasmodium, where they have been shown to be involved in cation homeostasis, polyphosphate metabolism, and osmoregulation. Their structural counterparts in the apicomplexan parasite Eimeria have not been fully characterized. In this work, the ultrastructural and chemical properties of acidocalcisomes in Eimeria were characterized. Electron microscopy analysis of Eimeria parasites showed the dense organelles called volutin granules similar to acidocalcisomes. Immunolocalization of the vacuolar proton pyrophosphatase, considered as a marker for acidocalcisomes, showed labeling in vesicles of size and distribution similar to the dense organelles seen by electron microscopy. Spectrophotometric measurements of the kinetics of proton uptake showed a vacuolar proton pyrophosphatase activity. X-ray mapping revealed significant amounts of Na, Mg, P, K, Ca, and Zn in their matrix. The results suggest that volutin granules of Eimeria parasites are acidic, dense organelles and possess structural and chemical properties analogous to those of other acidocalcisomes, suggesting a similar functional role in these parasites. PMID:21699625

  15. Efficacy of treatments with toltrazuril 7.5% and lasalocid sodium in sheep naturally infected with Eimeria spp.

    Science.gov (United States)

    Rodrigues, Fernando de Souza; Tavares, Luiz Eduardo Roland; Paiva, Fernando

    2016-01-01

    The objective of this study was to evaluate the efficacy of an experimental formulation of toltrazuril 7.5% + Trimix™ on a naturally acquired infection of Eimeria spp. in suckling lambs kept on pasture and, in another trial, evaluate the comparative efficacy between lasalocid and toltrazuril 7.5% + Trimix™ in newly weaned sheep under feedlot conditions that had been naturally infected with Eimeria spp. In the first experiment, 30 suckling lambs were divided into two groups: A - treated with toltrazuril 7.5% + Trimix™ and B- control. In experiment 2, 30 weaned sheep were divided into three groups: I - treated with toltrazuril 7.5% + Trimix™, II - treated with lasalocid and III - control. Treatment group A showed an efficacy of 90, 99.4 and 87.3% on days 5, 10 and 20, respectively. Treatment group I had an efficacy of 98.2, 92.6 and 94.5%, while group II had an efficacy of 72.7, 81.6 and 95.9% on days 7, 21 and 42, respectively. Eight Eimeria species were identified; E. ovinoidalis was the most common. Treatment with the toltrazuril 7.5% +Trimix ™ formulation was effective against Eimeria spp. in suckling lambs in field conditions and lambs weaned in under feedlot conditions.

  16. A simple, one-tube assay for the simultaneous detection and diagnosis of ten Australian poultry Eimeria.

    Science.gov (United States)

    Godwin, Rosamond M; Morgan, Jess A T

    2014-02-01

    Coccidiosis is a costly worldwide enteric disease of chickens caused by parasites of the genus Eimeria. At present, there are seven described species that occur globally and a further three undescribed, operational taxonomic units (OTUs X, Y, and Z) that are known to infect chickens from Australia. Species of Eimeria have both overlapping morphology and pathology and frequently occur as mixed-species infections. This makes definitive diagnosis with currently available tests difficult and, to date, there is no test for the detection of the three OTUs. This paper describes the development of a PCR-based assay that is capable of detecting all ten species of Eimeria, including OTUs X, Y, and Z in field samples. The assay is based on a single set of generic primers that amplifies a single diagnostic fragment from the mitochondrial genome of each species. This one-tube assay is simple, low-cost, and has the capacity to be high throughput. It will therefore be of great benefit to the poultry industry for Eimeria detection and control, and the confirmation of identity and purity of vaccine strains. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Incorporation of a recombinant Eimeria maxima IMP1 antigen into nanoparticles confers protective immunity against E. maxima challenge infection

    Science.gov (United States)

    The purpose of this study was to determine if incorporating a recombinant Eimeria maxima protein, namely rEmaxIMP1, into gold nanoparticles (NP) could improve the level of protective immunity against E. maxima challenge infection. Recombinant EmaxIMP1 was expressed in Escherchia coli as a poly-His f...

  18. Occurrence of Eimeria species parasites on small-scale commercial chicken farms in Africa and indication of economic profitability.

    Science.gov (United States)

    Fornace, Kimberly M; Clark, Emily L; Macdonald, Sarah E; Namangala, Boniface; Karimuribo, Esron; Awuni, Joseph A; Thieme, Olaf; Blake, Damer P; Rushton, Jonathan

    2013-01-01

    Small-scale commercial poultry production is emerging as an important form of livestock production in Africa, providing sources of income and animal protein to many poor households, yet the occurrence and impact of coccidiosis on this relatively new production system remains unknown. The primary objective of this study was to examine Eimeria parasite occurrence on small-scale commercial poultry farms in Ghana, Tanzania and Zambia. Additionally, farm economic viability was measured by calculating the farm gross margin and enterprise budget. Using these economic measures as global assessments of farm productivity, encompassing the diversity present in regional husbandry systems with a measure of fundamental local relevance, we investigated the detection of specific Eimeria species as indicators of farm profitability. Faecal samples and data on production parameters were collected from small-scale (less than 2,000 birds per batch) intensive broiler and layer farms in peri-urban Ghana, Tanzania and Zambia. All seven Eimeria species recognised to infect the chicken were detected in each country. Furthermore, two of the three genetic variants (operational taxonomic units) identified previously in Australia have been described outside of Australia for the first time. Detection of the most pathogenic Eimeria species associated with decreased farm profitability and may be considered as an indicator of likely farm performance. While a causal link remains to be demonstrated, the presence of highly pathogenic enteric parasites may pose a threat to profitable, sustainable small-scale poultry enterprises in Africa.

  19. A new species of Eimeria Schneider, 1875 (Apicomplexa: Eimeriidae) from Alectoris barbara (Aves: Phasianidae) from the Canary Islands (Spain)

    Czech Academy of Sciences Publication Activity Database

    Fernández-Alvarez, A.; Modrý, David; Foronda, P.

    2016-01-01

    Roč. 115, č. 5 (2016), s. 1817-1825 ISSN 0932-0113 Institutional support: RVO:60077344 Keywords : Coccidia * Eimeria barbarae n. sp * Alectoris barbara * Canary Island Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.329, year: 2016

  20. Two new species of Eimeria (Apicomplexa: Eimeriidae) from Asian geoemydid turtles Kachuga tentoria and Melanochelys trijuga (Testudines: Geoemydidae)

    Czech Academy of Sciences Publication Activity Database

    Široký, P.; Modrý, David

    2005-01-01

    Roč. 12, č. 1 (2005), s. 9-13 ISSN 1252-607X R&D Projects: GA ČR GP524/03/D104 Institutional research plan: CEZ:AV0Z60220518 Keywords : Eimeria * Apicomplexa * Testudines Subject RIV: EG - Zoology Impact factor: 0.844, year: 2005

  1. HIGH HYDROSTATIC PRESSURE AND UV LIGHT TREATMENT OF PRODUCE CONTAMINATED WITH EIMERIA ACERVULINA AS A CYCLOSPORA CAYETANENSIS SURROGATE

    Science.gov (United States)

    The prevalence, size, genome, and life cycle of Eimeria acervulina make this organism a good surrogate for Cyclospora cayetanensis, a protozoan that causes gastroenteritis in humans, including recent outbreaks in the United States and Canada associated with contaminated raspberries and basil. Labora...

  2. The rabbit coccidium Eimeria flavescens Marotel and Guilhon, 1941: an electron microscopic study of its life cycle

    Czech Academy of Sciences Publication Activity Database

    Pakandl, Michal; Černík, F.; Coudert, P.

    2003-01-01

    Roč. 91, č. 4 (2003), s. 304-311 ISSN 0932-0113 R&D Projects: GA AV ČR IBS6022002 Institutional research plan: CEZ:AV0Z6022909 Keywords : Coccidia * life cycle * Eimeria flavescens Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 1.000, year: 2003

  3. Development of molecular assays for the identification of the 11 Eimeria species of the domestic rabbit (Oryctolagus cuniculus)

    Czech Academy of Sciences Publication Activity Database

    Oliveira, U. C.; Fraga, J. S.; Licois, D.; Pakandl, Michal; Gruber, A.

    2011-01-01

    Roč. 176, 2/3 (2011), 275-280 ISSN 0304-4017 Institutional research plan: CEZ:AV0Z60220518 Keywords : Eimeria * Rabbit * Coccidiosis * ITS1 * Ribosomal DNA * PCR-based diagnosis Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.579, year: 2011

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

    Science.gov (United States)

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

    2013-09-03

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

  5. Experimental Infection with Sporulated Oocysts of Eimeria maxima (Apicomplexa: Eimeriidae) in Broiler

    Science.gov (United States)

    Brito, Luciana da S.; Pereira, Elder N.; da Silva, Augusta A.; Bentivóglio Costa Silva, Vinícius; Freitas, Fagner L. da C.

    2014-01-01

    Through this study we assessed the metabolic and pathological changes in broilers experimentally infected with oocysts of Eimeria maxima. To perform the experiment, we used 150 broiler strain cooB males, with ten days of age, were randomized according to weight and randomly assigned to two experimental groups: the control group was inoculated with 0.5 mL of distilled water; the infected group inoculated with 0.5 mL of solution containing 5 × 104 sporulated oocysts of Eimeria maxima. The live performance was evaluated on day 0 (day of inoculation), 5°, 10°, 15°, 25°, and 35° dpi, being slaughtered by cervical dislocation, fifteen birds/group. Although the sum in meat production was higher in the control group, the weight of the heart and gizzard of the experimental animals showed no significant difference, while the liver had difference on day 5°, 15°, and 35° dpi. The pathologic evaluation showed congested mucosa and presence of large amounts of mucus at 6 dpi. Therefore, it is concluded that the dose of 5 × 104  E. maxima inoculated in the experimental group was enough to cause harm to the animal organism. PMID:26464925

  6. Metabolic alterations in broiler chickens experimentally infected with sporulated oocysts of Eimeria maxima.

    Science.gov (United States)

    Freitas, Fagner Luiz da Costa

    2014-01-01

    Metabolic and morphometric alterations of the duodenal villi caused by parasitism of chickens by Eimeria maxima were evaluated, using 100 male Cobb birds, randomly distributed into two groups (control and infected). The infected group was inoculated with 0.5 ml of a solution containing 5 × 10³ sporulated oocysts of Eimeria maxima. Ten birds per sample were sacrificed on the 6th, 11th, 22nd and 41st days post-infection (dpi). In order to evaluate the alterations, samples of duodenum, jejunum and ileum fragments were collected after necropsy for histological analysis. Villus biometry was determined by means of a slide graduated in microns that was attached to a binocular microscope. To evaluate the biochemical data, 5 ml of blood were sampled from the birds before sacrifice. The statistical analyses were performed using the GraphPad 5 statistical software for Windows. Tukey's multiple comparison test (p maxima causes both qualitative and quantitative alterations to the structure of the intestinal villi, thereby interfering with the absorption of nutrients such as calcium, phosphorus, magnesium, protein and lipids, with consequent reductions in the birds' weights.

  7. Use of fluorescent nanoparticles to investigate nutrient acquisition by developing Eimeria maxima macrogametocytes.

    Science.gov (United States)

    Frölich, Sonja; Wallach, Michael

    2016-06-29

    The enteric disease coccidiosis, caused by the unicellular parasite Eimeria, is a major and reoccurring problem for the poultry industry. While the molecular machinery driving host cell invasion and oocyst wall formation has been well documented in Eimeria, relatively little is known about the host cell modifications which lead to acquisition of nutrients and parasite growth. In order to understand the mechanism(s) by which nutrients are acquired by developing intracellular gametocytes and oocysts, we have performed uptake experiments using polystyrene nanoparticles (NPs) of 40 nm and 100 nm in size, as model NPs typical of organic macromolecules. Cytochalasin D and nocodazole were used to inhibit, respectively, the polymerization of the actin and microtubules. The results indicated that NPs entered the parasite at all stages of macrogametocyte development and early oocyst maturation via an active energy dependent process. Interestingly, the smaller NPs were found throughout the parasite cytoplasm, while the larger NPs were mainly localised to the lumen of large type 1 wall forming body organelles. NP uptake was reduced after microfilament disruption and treatment with nocodazole. These observations suggest that E. maxima parasites utilize at least 2 or more uptake pathways to internalize exogenous material during the sexual stages of development.

  8. High pathogenicity and strong immunogenicity of a Chinese isolate of Eimeria magna Pérard, 1925.

    Science.gov (United States)

    Tao, Geru; Wang, Yunzhou; Li, Chao; Gu, Xiaolong; Cui, Ping; Fang, Sufang; Suo, Xun; Liu, Xianyong

    2017-06-01

    Coccidia infection of rabbits with one or several species of parasites of the genus Eimeria causes coccidiosis, a disease leading to huge economic losses in the rabbit industry. Eimeria magna, one of the causal agents of rabbit coccidiosis, was characterized as mildly pathogenic and moderately immunogenic in previous studies. In this study, we identified a Chinese isolate of E. magna by testing its biological features (oocyst morphology and size, prepatent time) and sequencing its internal transcribed spacer 1 (ITS-1) DNA fragment. This isolate is highly pathogenic; infection of rabbits with only 1×10 2 oocysts caused a 55% reduction in weight gain in 14days. In addition, immunization with 1×10 2 oocysts prevented body weight loss against re-infection with 5×10 4 oocysts, indicating the high immunogenicity of this isolate. Our study described the distinctive phenotype of the Chinese isolate of E. magna and contributed to the research of geographic variation of rabbit coccidia. Copyright © 2017 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2014-09-01

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

  10. Humoral immune responses of experimentally Eimeria ninakholyakimovae-infected goat kids.

    Science.gov (United States)

    Matos, Lorena; Muñoz, María Del Carmen; Molina, José Manuel; Ferrer, Otilia; Rodríguez, Francisco; Pérez, Davinia; López, Adassa María; Martín, Sergio; Hermosilla, Carlos; Taubert, Anja; Ruiz, Antonio

    2017-04-01

    Although cellular immune reactions seem to be crucial for protective immune responses in Eimeria spp. infections, there are also evidences on an active involvement of the humoral counterpart. In the present study, we have analyzed the humoral response of goat kids subjected to primary and challenge infections with Eimeria ninakholyakimovae. Specific levels of IgG and IgM in serum samples and IgA in the ileal mucus were estimated. In infected kids, significantly increased levels of IgG were observed from 3 weeks post infection onwards in addition to an enhancement of specific IgM and secretory IgA levels. A wide range of peptides of sporulated oocyst antigen (SOA) was recognized by specific IgG as determined by immunoblotting. However, no correlations were found between immunoglobulin levels and OPG counts after challenge infection. Overall, these data indicate a significant specific humoral response of E. ninakohlyakimovae-infected goat kids that does not seem to convey immunoprotection. Further studies should be addressed to clarify if the lack of correlation might be associated to the type of antigen used for the immunoenzimatic assays, the age of the animals or other factors. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. [Species of the genus Eimeria (Apicomplexa: Eimeriidae) in giant anteaters (Myrmecophaga tridactyla Linnaeus, 1758) in captivity].

    Science.gov (United States)

    Freitas, Fagner Luiz da C; Almeida, Katyane de S; Zanetti, André S; do Nascimento, Adjair A; Machado, Cé Lio R; Machado, Rosangela Z

    2006-01-01

    The parasitism of the two giant anteaters adults (Myrmecophaga tridactyla), one male and one female, infected naturally with Eimeria escomeli, E. tamanduae e E. marajoensis was related in the present research. In E. escomeli oocysts were 23.9 +/- 1.89 by 19.7 +/- 1.60 microm and its sporocysts were 11.47 +/- 1.25 by 6.48 +/- 0.80 microm. In E. tamanduae oocysts were 23.52 +/- 0.95 by 20.59 +/- 0.92 microm and its sporocysts were 12.19 +/- 0.65 by 7.15 +/- 0.55 microm. In E. marajoensis oocysts were 13.5 +/- 1.7 by 13.1 +/- 1.8 microm and its sporocysts were 7.4 +/- 0.58 by 5.4 +/- 0.8 microm. Eimeria escomeli was described before parasitizing giants anteater from Bolivia, and it was point out as the first time in Brazil. The presence of E. tamanduae and E. marajoensis parasitizing giant anteaters indicate the possibility of having co-infection of them among animals of the family Myrmecophagidae.

  12. IDENTIFICATION AND CHARACTERIZATION OF Eimeria spp. DURING EARLY NATURAL INFECTION IN GOAT KIDS IN BAJA CALIFORNIA SUR, MEXICO

    Directory of Open Access Journals (Sweden)

    Ramón Cepeda-Palacios

    2015-12-01

    Full Text Available The aim of this study was to characterize the natural infection by Eimeria spp. in goat kids, and to describe some pathophysiological responses to eimerosis in kids under intensive rearing conditions in B.C.S, Mexico. Nineteen adult crossbred does naturally infected with mixed Eimeria spp. and 20 Anglo Nubian x Creole crossbred kids were used. Oocyst per gram of feces (OPG and identification of Eimeria species were determined in does (during the pre-kidding and post-kidding periods and kids. Clinical signs, hematocrit, hemoglobin and alkaline phosphatase activity in blood serum were evaluated. OPG (mean±SD was significantly higher (P<0.05 in pre-kidding (9,478±7,599 than in post-kidding (5,313±2,909 period. Oocyst elimination in feces began at age 59±9 days in kids. Eimerian species identified were E. arloingi, E. jolchijevi, E. ninakohlyakimovae, E. hirci, E. christenseni and E. alijevi. Kids were humanely sacrificed to evaluate pathological lesions. Intestinal lesions and lesion severity showed differences in duodenum, jejunum, ileum, cecum and colon, being more severe in duodenum. In conclusion, OPG increased during the late pregnancy in does which favored a doe-kid transmission mechanism. Our results support the notion of Eimeria reproduction rhythms during the late pregnancy period in goats, and this reproduction contribute to vertical transmission of Eimeria to the newborn. However, coccidian outbreaks are developed and clinically observed only when stressing factors such as when weaning occur. Coccidia had devastating effects on the intestine of kids, which might cause long-term permanent malabsortion consequences. Â

  13. Relationship Between Broiler Body Weights, Eimeria maxima Gross Lesion Scores, and Microscores in Three Anticoccidial Sensitivity Tests.

    Science.gov (United States)

    Barrios, Miguel A; Da Costa, Manuel; Kimminau, Emily; Fuller, Lorraine; Clark, Steven; Pesti, Gene; Beckstead, Robert

    2017-06-01

    Anticoccidial sensitivity tests (ASTs) serve to determine the efficacy of anticoccidial drugs against Eimeria field isolates in a controlled laboratory setting. The most commonly measured parameters are body weight gain, feed conversion ratio, gross intestinal lesion scores, and mortality. Due to the difficulty in reliably scoring gross lesion scores of Eimeria maxima , microscopic analysis of intestinal scrapings (microscores) can be used in the field to indicate the presence of this particular Eimeria. The goal of this study was to determine the relationship between E. maxima microscores and broiler body weights and gross E. maxima lesion scores in three ASTs. Day-old broiler chicks were raised for 12 days on a standard corn-soy diet. On Day 12, chicks were placed in Petersime batteries and treatment diets were provided. There were six birds per pen, four pens per treatment, and 12 treatments, for a total of 288 chicks per AST. The treatments were as follows: 1) nonmedicated, noninfected; 2) nonmedicated, infected; 3) lasalocid, infected; 4) salinomycin, infected; 5) diclazuril, infected; 6) monensin, infected; 7) decoquinate, infected; 8) narasin + nicarbazin, infected; 9) narasin, infected; 10) nicarbazin, infected; 11) robenidine, infected; and 12) zoalene, infected. On Day 14, chicks were challenged with an Eimeria field isolate by oral gavage. On Day 20, broilers were weighed, and gross lesion scores and microscores were classified from 0 to 4 depending on the severity of the gross lesion scores and E. maxima microscores. Data from three trials using different field isolates were statistically analyzed using a logarithmic regression model. There was no relationship (P = 0.1224) between microscores and body weight gain. There was a positive relationship between microscores and gross lesion scores (P = 0.004). However, there was also an interaction between isolate and treatment (P Eimeria or the amount of E. maxima in the inoculum.

  14. Differential responses of cecal microbiota to fishmeal, Eimeria and Clostridium perfringens in a necrotic enteritis challenge model in chickens.

    Directory of Open Access Journals (Sweden)

    Dragana Stanley

    Full Text Available Clostridium perfringens causes enteric diseases in animals and humans. In poultry, avian-specific C. perfringens strains cause necrotic enteritis, an economically significant poultry disease that costs the global industry over $2 billion annually in losses and control measures. With removal of antibiotic growth promoters in some countries this disease appears to be on the rise. In experimental conditions used to study disease pathogenesis and potential control measures, reproduction of the disease relies on the use of predisposing factors such as Eimeria infection and the use of high protein diets, indicating complex mechanisms involved in the onset of necrotic enteritis. The mechanisms by which the predisposing factors contribute to disease progression are not well understood but it has been suggested that they may cause perturbations in the microbiota within the gastrointestinal tract. We inspected changes in cecal microbiota and short chain fatty acids (SCFA induced by Eimeria and fishmeal, in birds challenged or not challenged with C. perfringens. C. perfringens challenge in the absence of predisposing factors did not cause significant changes in either the alpha or beta diversity of the microbiota nor in concentrations of SCFA. Moreover, there was no C. perfringens detected in the cecal microbiota 2 days post-challenge without the presence of predisposing factors. In contrast, both fishmeal and Eimeria caused significant changes in microbiota, seen in both alpha and beta diversity and also enabled C. perfringens to establish itself post challenge. Eimeria had its strongest influence on intestinal microbiota and SCFA when combined with fishmeal. Out of 6 SCFAs measured, including butyric acid, none were significantly influenced by C. perfringens, but their levels were strongly modified following the use of both predisposing factors. There was little overlap in the changes caused following Eimeria and fishmeal treatments, possibly indicating

  15. Molecular and phylogenetic characterizations of an Eimeria krijgsmanni Yakimoff & Gouseff, 1938 (Apicomplexa: Eimeriidae) mouse intestinal protozoan parasite by partial 18S ribosomal RNA gene sequence analysis.

    Science.gov (United States)

    Takeo, Toshinori; Tanaka, Tetsuya; Matsubayashi, Makoto; Maeda, Hiroki; Kusakisako, Kodai; Matsui, Toshihiro; Mochizuki, Masami; Matsuo, Tomohide

    2014-08-01

    Previously, we characterized an undocumented strain of Eimeria krijgsmanni by morphological and biological features. Here, we present a detailed molecular phylogenetic analysis of this organism. Namely, 18S ribosomal RNA gene (rDNA) sequences of E. krijgsmanni were analyzed to incorporate this species into a comprehensive Eimeria phylogeny. As a result, partial 18S rDNA sequence from E. krijgsmanni was successfully determined, and two different types, Type A and Type B, that differed by 1 base pair were identified. E. krijgsmanni was originally isolated from a single oocyst, and thus the result show that the two types might have allelic sequence heterogeneity in the 18S rDNA. Based on phylogenetic analyses, the two types of E. krijgsmanni 18S rDNA formed one of two clades among murine Eimeria spp.; these Eimeria clades reflected morphological similarity among the Eimeria spp. This is the third molecular phylogenetic characterization of a murine Eimeria spp. in addition to E. falciformis and E. papillata. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  16. The SnSAG merozoite surface antigens of Sarcocystis neurona are expressed differentially during the bradyzoite and sporozoite life cycle stages.

    Science.gov (United States)

    Gautam, A; Dubey, J P; Saville, W J; Howe, D K

    2011-12-29

    Sarcocystis neurona is a two-host coccidian parasite whose complex life cycle progresses through multiple developmental stages differing at morphological and molecular levels. The S. neurona merozoite surface is covered by multiple, related glycosylphosphatidylinositol-linked proteins, which are orthologous to the surface antigen (SAG)/SAG1-related sequence (SRS) gene family of Toxoplasma gondii. Expression of the SAG/SRS proteins in T. gondii and another related parasite Neospora caninum is life-cycle stage specific and seems necessary for parasite transmission and persistence of infection. In the present study, the expression of S. neurona merozoite surface antigens (SnSAGs) was evaluated in the sporozoite and bradyzoite stages. Western blot analysis was used to compare SnSAG expression in merozoites versus sporozoites, while immunocytochemistry was performed to examine expression of the SnSAGs in merozoites versus bradyzoites. These analyses revealed that SnSAG2, SnSAG3 and SnSAG4 are expressed in sporozoites, while SnSAG5 was appeared to be downregulated in this life cycle stage. In S. neurona bradyzoites, it was found that SnSAG2, SnSAG3, SnSAG4 and SnSAG5 were either absent or expression was greatly reduced. As shown for T. gondii, stage-specific expression of the SnSAGs may be important for the parasite to progress through its developmental stages and complete its life cycle successfully. Thus, it is possible that the SAG switching mechanism by these parasites could be exploited as a point of intervention. As well, the alterations in surface antigen expression during different life cycle stages may need to be considered when designing prospective approaches for protective vaccination. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. Co-localization of a CD1d-binding glycolipid with a radiation-attenuated sporozoite vaccine in LN-resident DCs for a robust adjuvant effect

    OpenAIRE

    Li, Xiangming; Kawamura, Akira; Andrews, Chasity D.; Miller, Jessica L.; Wu, Douglass; Tsao, Tiffany; Zhang, Min; Oren, Deena; Padte, Neal N.; Porcelli, Steven A.; Wong, Chi-Huey; Kappe, Stefan H. I.; Ho, David D.; Tsuji, Moriya

    2015-01-01

    A CD1d-binding glycolipid, α-Galactosylceramide (αGalCer), activates invariant natural killer T (iNKT) cells and acts as an adjuvant. We previously identified a fluorinated phenyl ring-modified αGalCer analog, 7DW8-5, displaying nearly 100-fold stronger CD1d binding affinity. In the present study, 7DW8-5 was found to exert a more potent adjuvant effect than αGalCer for a vaccine based on radiation-attenuated sporozoites (RAS) of a rodent malaria parasite, Plasmodium yoelii, also referred to a...

  18. Coccidies genus Eimeria as bioindicator of radioactive pollution of the biocenoses

    International Nuclear Information System (INIS)

    Pel'gunov, A.N.

    2006-01-01

    The effect of ecosystems contamination on morphological changes and sporulation of oocytes of Coccidae r. Eimeria, which are rodent parasites, is analyzed. Coccidae of rodents were collected in Chernobyl (1989-1991) and the Bryansk region of Russia (1992-1999). The surface contamination of experimental plots was varied from 0.11 to 11.8 MBq/m 2 . It was found that parametric sings (size and form index of oocytes) were independent from contamination level. Whereas nonparametric sings (deformations of oocytes shells and internal structure) and sporulatio process depend on contamination levels. The part of nonsporulated oocytes increased with contamination increasing but the the quantity of oocytes corresponding to the description of given type (norm) decreased. The dependence is well described by the logarithmic regression equation that allows to use these indexes for biological indication of ecosystem contamination [ru

  19. The economic impact of infection with Eimeria spp. in broiler farms from Romania

    Directory of Open Access Journals (Sweden)

    Adriana Györke

    2016-05-01

    Full Text Available ABSTRACT A survey was conducted on chicken broiler farms from Romania in August-November 2010 to evaluate economic losses due to coccidiosis. Data were collected from six broiler farms of different capacity regarding chemoprophylaxis program, weight gain, feed conversion, and mortality, for two previous flocks in two houses of each farm, and finally we evaluated the economic losses. Also, faeces samples were collected and oocysts were classified according to their size, and virulence of each Eimeria spp. field isolate was determined by lesion scoring. Correlations between economic performance, oocysts category, and virulence of Eimeria were assessed by multiple linear regression. Total economic losses per 24 flocks of 18,000 chicks each were about €37,948.2, with an average of €3,162.4 per flock, and they were caused by mortality (34.8% and poor feed conversion (65.2%. Poor body weight gain was associated with AM oocyst category (presumptively E. acervulina and/or E. mitis, high lesion score in the duodenum, and coccidiostat used for chemoprophylaxis. Feed conversion ratio was linked to the same parameters as body weight gain, minus chemoprophylaxis programme, plus total lesion score. The percentage of mortality was influenced by the lesion score in the caecum and total lesion score. Statistical analysis showed that epidemiological survey of broiler flocks during the grower period can help the farmer to avoid important economic losses due to coccidiosis. As in other countries, the economic losses caused by coccidiosis in Romania are important, and a good prophylaxis programme can reduce the economic impact of coccidiosis.

  20. Interactive effects of dietary arginine and Eimeria acervulina infection on broiler growth performance and metabolism.

    Science.gov (United States)

    Rochell, S J; Helmbrecht, A; Parsons, C M; Dilger, R N

    2017-03-01

    The influence of dietary Arg concentration and Eimeria acervulina infection on broiler growth performance and plasma carotenoid, nitric oxide (NO), amino acid, and urea concentrations was evaluated. Male Ross × Ross 308 broilers (384 total) were fed a common diet for 10 d post-hatch and provided experimental diets formulated to contain 1.23 (HA) or 0.74% (LA) standardized ileal digestible Arg from 10 to 28 d. At 21 d, one-half of the broilers were switched to the opposite diet to create 4 dietary regimens where birds were fed the LA diet throughout, the LA diet replaced by the HA diet at 21 d, the HA diet throughout, or the HA diet replaced by the LA diet at 21 d. Broilers were orally inoculated 0 (uninfected) or 3.5 × 105 sporulated E. acervulina oocysts at 15 d, resulting in a factorial arrangement of 4 dietary regimens × 2 infection states (8 replicates/treatment). Overall (10 to 28 d) BW gain and G:F were greatest (P Eimeria acervulina infection decreased (P  0.05) of E. acervulina on BW gain or G:F of broilers from 21 to 28 d. Plasma Arg, Lys, and Orn levels at 21 d indicated that the LA diet caused an imbalance in the Lys and Arg status of broilers, and E. acervulina infection increased (P < 0.01) the plasma concentration of these 3 amino acids. Diet × infection interactions (P < 0.05) were observed on 21 d for plasma carotenoids and NO, whereby infection decreased plasma carotenoids and increased plasma NO, but dietary Arg concentration only influenced these measures for uninfected birds. Thus, production of NO during E. acervulina infection was not impaired by dietary Arg limitation. © 2016 Poultry Science Association Inc.

  1. Growth performance and gastrointestinal responses of broiler chickens fed corn-soybean meal diet without or with exogenous epidermal growth factor upon challenge with Eimeria1

    Science.gov (United States)

    Kim, E.; Leung, H.; Akhtar, N.; Li, J.; Barta, J. R.; Wang, Y.; Yang, C.; Kiarie, E.

    2017-01-01

    Abstract Epidermal growth factor (EGF), a protein known for its mitogenic and anti-apoptotic effects was fed to broiler chickens to evaluate growth performance, gastrointestinal measurements, and apparent retention (AR) of components upon challenge with Eimeria. A total of 216, d old male broiler chicks (Ross 708) were placed in cages (6 birds/cage) and allocated to treatments. The treatments were: 1) control (Lactotobacilli lactis fermentation supernatant without EGF), 2) 80 μg of EGF/kg BW/d, and 3) 160 μg of EGF/kg BW/d. A basal antibiotic-free corn-soybean diet containing TiO2 was used. Birds were offered fresh feed with respective treatments on daily basis and had free access to drinking water for 14 d. On d 5, birds (6 replicates per treatment) were challenged with 1 mL of E. acervulina and E. maxima mixture via oral gavage and the other 6 replicates were given sham. Growth performance was measured in pre- (d 0 to 5) and post- (d 6 to 14) challenge periods. Two birds per cage were necropsied on d 10 for intestinal lesion scores and tissue samples for histomorphology and expression of select intestinal genes. Excreta samples for AR of components and oocyst shedding were taken d 10 to 13 and all birds were necropsied on d 14 for gastrointestinal weight. The EGF linearly (P Eimeria interaction (P > 0.05) on growth performance, AR of GE, and intestinal histomorphology; the main effects were such that Eimeria depressed (P Eimeria (P Eimeria challenged birds whilst no effect in non-challenged control. In conclusion, Eimeria challenge reduced growth performance and impaired gut function; EGF showed beneficial effects on growth pre-challenge and improved indices of gut function upon Eimeria challenge. PMID:28938785

  2. 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. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Growth performance and gastrointestinal responses of broiler chickens fed corn-soybean meal diet without or with exogenous epidermal growth factor upon challenge with Eimeria.

    Science.gov (United States)

    Kim, E; Leung, H; Akhtar, N; Li, J; Barta, J R; Wang, Y; Yang, C; Kiarie, E

    2017-10-01

    Epidermal growth factor (EGF), a protein known for its mitogenic and anti-apoptotic effects was fed to broiler chickens to evaluate growth performance, gastrointestinal measurements, and apparent retention (AR) of components upon challenge with Eimeria. A total of 216, d old male broiler chicks (Ross 708) were placed in cages (6 birds/cage) and allocated to treatments. The treatments were: 1) control (Lactotobacilli lactis fermentation supernatant without EGF), 2) 80 μg of EGF/kg BW/d, and 3) 160 μg of EGF/kg BW/d. A basal antibiotic-free corn-soybean diet containing TiO2 was used. Birds were offered fresh feed with respective treatments on daily basis and had free access to drinking water for 14 d. On d 5, birds (6 replicates per treatment) were challenged with 1 mL of E. acervulina and E. maxima mixture via oral gavage and the other 6 replicates were given sham. Growth performance was measured in pre- (d 0 to 5) and post- (d 6 to 14) challenge periods. Two birds per cage were necropsied on d 10 for intestinal lesion scores and tissue samples for histomorphology and expression of select intestinal genes. Excreta samples for AR of components and oocyst shedding were taken d 10 to 13 and all birds were necropsied on d 14 for gastrointestinal weight. The EGF linearly (P Eimeria interaction (P > 0.05) on growth performance, AR of GE, and intestinal histomorphology; the main effects were such that Eimeria depressed (P Eimeria (P Eimeria challenged birds whilst no effect in non-challenged control. In conclusion, Eimeria challenge reduced growth performance and impaired gut function; EGF showed beneficial effects on growth pre-challenge and improved indices of gut function upon Eimeria challenge. © The Author 2017. Published by Oxford University Press on behalf of Poultry Science Association.

  4. Control of eight predominant Eimeria spp. involved in economic coccidiosis of broiler chicken by a chemically characterized essential oil.

    Science.gov (United States)

    Barbour, E K; Bragg, R R; Karrouf, G; Iyer, A; Azhar, E; Harakeh, S; Kumosani, T

    2015-03-01

    To control eight most predominant Eimeria spp. involved in the economic disease of coccidiosis in broiler chicken, by a chemically characterized essential oil of eucalyptus and peppermint. The experimental design consisted of 160 day-old-broiler chicks, divided into four equal groups (G1 , G2 , G3 and G4 ), with 40 birds per group. Each group was divided into four equal subgroups. Birds in G1 were deprived of essential oil treatment and of Eimeria challenge. Birds in G2 were unchallenged, and administered the essential oil in drinking water at 0.69 ml kg(-1) body weight. Birds in G3 were untreated with essential oil, and each of its four subgroups was challenged at a different age (14, 21, 28 and 35 days). Birds in G4 were treated with essential oil, and challenged in the same manner as for G3 . Equal number of birds from all subgroups (n = 10) were sacrificed at the sixth day after the time allocated for each challenge. The 6 day incubation period post challenge resulted in respective mean per cent weight increase in G2 and G1 birds equivalent to 57.8 and 53.1% (P essential oil improved the per cent weight increase in challenged birds (54.6%) compared to the challenged-untreated birds (18.6%) (P essential oils of eucalyptus and peppermint to control the most prevalent Eimeria spp. involved in coccidiosis of broiler chicken, helping in improvement of their production, alleviation of lesions and reduction in intestinal oocyst counts. This study provides information about the possibility of using this blend of essential oil as a coccidiostat for the protection of broiler chickens against the prevalent eight Eimeria spp. of coccidiosis. © 2014 The Society for Applied Microbiology.

  5. Eimeria lokuma n. sp. (Apicomplexa: Eimeriidae), a new coccidium from the African helmeted turtle Pelomedusa subrufa (Lacepede) (Testudines: Pelomedusidae)

    Czech Academy of Sciences Publication Activity Database

    Široký, P.; Kamler, M.; Modrý, David

    2006-01-01

    Roč. 65, č. 1 (2006), s. 73-76 ISSN 0165-5752 R&D Projects: GA ČR GD524/03/H133; GA ČR GP524/03/D104 Institutional research plan: CEZ:AV0Z60220518 Keywords : Eimeria * Pelomedusa * Apicomplexa Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 0.856, year: 2006

  6. 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.

  7. The Efficacies of Banana Stem Extract as a Candidate of Coccidiostat Against Rabbit Eimeria Stiedaio Ocysts: an in Vitro Analysis

    Directory of Open Access Journals (Sweden)

    Diana Indrasanti

    2015-09-01

    Full Text Available The objective of this research was to investigatethe ability of banana stem (Musa paradisiaca to inhibitsporulation of Eimeria stiedaioocystsderived fromrabbit by in vitroanalysis.Analyze the active substance proximate analysis and active substancesin this research were performed too. Banana stem extract were used in this experiment andsulfaquinoxalline(Coxy ®was run as acontrol. The Eimeria stiedaioocystswere incubated prior the presence of  different concentration from banana stem extract  0%, 1%, 2%, 4%, 8%for 1, 2 and 3 daysat 26°C. In addition,Factorial patterned Completely Randomized Design (CRD with five replicates wasapplied on the experiment. Result analysis was performed by using Analysis of Variance and following by Honestly Significant Difference (HSD post hoc test. Here, we identified that banana stem extract contain different type of active substance such as tannin, saponin, and alkaloid. Banana stem extract significantly affected the oocysts sporulation included the amount of sporulatedoocysts (P<0.01, unsporulatedoocysts (P<0.01, and transformed oocysts (P<0.01. In conclusion banana stem could inhibit the development of Eimeria stiedaioocysts on in vitroexperiment. HSD test showed that the optimum potential efficacy of banana stem toinhibit sporulation was at 4% and 8% concentration during three days incubation.

  8. The effect of Eimeria maxima infection on the expression of amino acid and sugar transporters aminopeptidase, as well as the di- and tri-peptide transporter PepT1, is not solely due to decreased feed intake

    Science.gov (United States)

    Coccidiosis caused by Eimeria in poultry is endemic to poultry operations and results in decreased feed intake, diarrhea, and decreased weight gain. The goal was to determine the effect infection Eimeria maxima on the expression of genes that encode peptide and amino acid transporters (AATs), and al...

  9. Estudos anatômicos de folhas de espécies de plantas daninhas de grande ocorrência no Brasil: IV - Amaranthus deflexus, Amaranthus spinosus, Alternanthera tenella e Euphorbia heterophylla Leaf anatomical studies in weed species widely common in Brazil: IV - Amaranthus deflexus, Amaranthus spinosus, Alternanthera tenella and Euphorbia heterophylla

    Directory of Open Access Journals (Sweden)

    E.A. Ferreira

    2003-08-01

    Full Text Available Neste trabalho é apresentado um estudo da anatomia das folhas de espécies de plantas daninhas de grande ocorrência no Brasil: Amaranthus deflexus, Amaranthus spinosus, Alternanthera tenella e Euphorbia heterophylla, visando melhor compreensão das barreiras que cada espécie impõe à penetração dos herbicidas e outros compostos utilizados em aplicações foliares. As folhas completamente expandidas do terceiro ao quinto nó foram coletadas de plantas de ocorrência espontânea no campo. Das folhas de cada espécie foram obtidas três amostras da região mediana, com aproximadamente 1 cm², as quais foram utilizadas em estudos da estrutura, clarificação e em observações em microscópio eletrônico de varredura. Todas as espécies avaliadas são anfiestomáticas. As principais barreiras foliares potenciais à penetração de herbicidas observadas nas plantas daninhas A. deflexus e A. spinosus foram, respectivamente, grande espessura da cutícula da face adaxial e da cutícula das duas faces. Já em relação a A. tenella, grande espessura da cutícula das duas faces, elevado teor de cera epicuticular e alta densidade tricomática foram os principais obstáculos potenciais detectados. E. heterophylla apresentou como possíveis principais barreiras foliares à penetração de agroquímicos o alto teor de cera epicuticular, a elevada densidade de laticíferos e a grande espessura da cutícula da face adaxial.This research aimed to study the leaf anatomy of the weed species Amaranthus deflexus, Amaranthus spinosus, Alternanthera tenella and Euphorbia heterophylla, widely known in Brazil, to acquire a better understanding of the barriers each species imposes to herbicide penetration, and to other substances used for leaf spraying. Completely expanded leaves from the third to the fifth nodes were collected from spontaneous plants in the field. Three samples approximately 1cm², were removed from the medium portion of the leaves, from each

  10. Reproductive effort and seasonality associated with male-biased parasitism in Gracilinanus agilis (Didelphimorphia: Didelphidae) infected by Eimeria spp. (Apicomplexa: Eimeriidae) in the Brazilian cerrado.

    Science.gov (United States)

    Strona, A L S; Levenhagem, M; Leiner, N O

    2015-07-01

    The aggregation of parasites among hosts is associated with differential host exposure and susceptibility to parasites, which varies according to host gender, body size, reproductive status and environmental factors. We evaluated the role of these factors on infestation by Eimeria spp. (Eimeriidae) in the agile gracile mouse opossum (Gracilinanus agilis), a semelparous didelphid inhabiting neotropical savannahs. Eimeria spp. abundance and prevalence among G. agilis were associated with the breeding status of individuals and to a lesser extent to climatic season, with both sexes presenting higher Eimeria spp. burdens during late breeding/wet season. On the other hand, male-biased parasitism was restricted to dry/mating season. We suggest that male spatial organization and diet may account for increased parasite burdens within this sex, although future studies should evaluate the role of physiological differences associated with androgen hormones. Finally, a rapid increase in Eimeria spp. loads among females during the late breeding/wet season seems associated with seasonal changes in susceptibility, due to breeding costs related to semelparity, and exposure to infective propagules, while male-die off seems to explain maintenance of higher Eimeria spp. burdens within this sex in the same period.

  11. 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...

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

    Science.gov (United States)

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

    2018-05-01

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

  13. Colocalization of a CD1d-Binding Glycolipid with a Radiation-Attenuated Sporozoite Vaccine in Lymph Node-Resident Dendritic Cells for a Robust Adjuvant Effect.

    Science.gov (United States)

    Li, Xiangming; Kawamura, Akira; Andrews, Chasity D; Miller, Jessica L; Wu, Douglass; Tsao, Tiffany; Zhang, Min; Oren, Deena; Padte, Neal N; Porcelli, Steven A; Wong, Chi-Huey; Kappe, Stefan H I; Ho, David D; Tsuji, Moriya

    2015-09-15

    A CD1d-binding glycolipid, α-Galactosylceramide (αGalCer), activates invariant NK T cells and acts as an adjuvant. We previously identified a fluorinated phenyl ring-modified αGalCer analog, 7DW8-5, displaying nearly 100-fold stronger CD1d binding affinity. In the current study, 7DW8-5 was found to exert a more potent adjuvant effect than αGalCer for a vaccine based on radiation-attenuated sporozoites of a rodent malaria parasite, Plasmodium yoelii, also referred to as irradiated P. yoelii sporozoites (IrPySpz). 7DW8-5 had a superb adjuvant effect only when the glycolipid and IrPySpz were conjointly administered i.m. Therefore, we evaluated the effect of distinctly different biodistribution patterns of αGalCer and 7DW8-5 on their respective adjuvant activities. Although both glycolipids induce a similar cytokine response in sera of mice injected i.v., after i.m. injection, αGalCer induces a systemic cytokine response, whereas 7DW8-5 is locally trapped by CD1d expressed by dendritic cells (DCs) in draining lymph nodes (dLNs). Moreover, the i.m. coadministration of 7DW8-5 with IrPySpz results in the recruitment of DCs to dLNs and the activation and maturation of DCs. These events cause the potent adjuvant effect of 7DW8-5, resulting in the enhancement of the CD8(+) T cell response induced by IrPySpz and, ultimately, improved protection against malaria. Our study is the first to show that the colocalization of a CD1d-binding invariant NK T cell-stimulatory glycolipid and a vaccine, like radiation-attenuated sporozoites, in dLN-resident DCs upon i.m. conjoint administration governs the potency of the adjuvant effect of the glycolipid. Copyright © 2015 by The American Association of Immunologists, Inc.

  14. Protection of Broiler Chicks Housed with Immunized Cohorts Against Infection with Eimeria maxima and E. acervulina.

    Science.gov (United States)

    Fetterer, Raymond H; Barfield, Ruth C; Jenkins, Mark C

    2015-03-01

    The use of live oocyst vaccines is becoming increasingly important in the control of avian coccidiosis in broilers. Knowledge of the mechanisms employed when chicks uptake oocysts and become immune is important for optimizing delivery of live vaccines. The current study tests the hypothesis that chicks not initially immunized may ingest oocysts by contact with litter containing oocysts shed by immunized cohorts. In Experiment 1, day-old broiler chicks were housed in pens containing clean litter. In Trial 1, 100% of chicks in some pens were immunized with 2.5 X 10(3) Eimeria acervulina oocysts while in other pens only 75% of chicks were immunized and remaining cohorts within the pens were not immunized. Other pens contained chicks that served as nonimmunized nonchallenged controls or nonimmunized challenged controls (NIC). On day 21, birds were given a homologous challenge of 6 X 10(5) oocysts. A second identical trial was conducted, except birds were immunized with 500 Eimeria maxima oocysts and were challenged with 3 X 10(3) E. maxima oocysts. In Experiment 2, 100% of chicks in some pens were immunized with 500 E. acervulina oocysts while in other pens either 75% or 50% of the birds were immunized. On day 14, birds were challenged with 1 X 10(6) oocysts. Trial 2 was identical to Trial 1 except that birds were immunized with 100 E. maxima oocysts and challenged with 1 X 10(6) oocysts. For all experiments weight gain, feed conversion ratio (FCR), plasma carotenoids, and litter oocyst counts were measured. In Experiment 1, the level of protection in groups containing 25% nonimmunized cohorts, as measured by weight gain, carotenoid level, FCR, and oocyst litter counts, was identical to groups containing 100% immunized chicks. In Experiment 2, pens where 50% or 75% of birds were immunized with either E. maxima or E. acervulina were not well protected from decreases in weight gain and plasma carotenoids nor from increases in litter oocyst counts following a challenge

  15. Protective immune responses during prepatency in goat kids experimentally infected with Eimeria ninakohlyakimovae.

    Science.gov (United States)

    Matos, L; Muñoz, M C; Molina, J M; Rodríguez, F; Perez, D; Lopez, A; Ferrer, O; Hermosilla, C; Taubert, A; Ruiz, A

    2017-08-15

    During the first schizogony, the goat coccidia Eimeria ninakohlyakimovae develops macroschizonts in lacteal duct endothelial cells, whose rupture leads to severe ileal damage and clinical signs during the prepatent period. The immune response elicited against early stages of the parasite development still requires to be investigated. In the present study we have evaluated immune reactions in goat kids primary- and challenged-infected with Eimeria ninakohlyakimovae, and sacrificed during prepatency (7days after challenge). The oocyst output during the primary infection, body weight and clinical condition of all the animals were examined and, at the end of the experiment, all the goat kids were euthanized and subjected to necropsy. Samples were taken from different sections of the ileum, colon and mesenteric lymph nodes (MLN) of primary- and challenged E. ninakohlyakimovae-infected animals. Intestinal leukocyte subpopulations were characterized in E. ninakohlyakimovae-infected mucosa and counts of lymphocytes, eosinophils, polymorphonuclear neutrophils (PMN), globular leukocytes and mast cells were recorded. Additionally, gene expression of caprine IL-2, IL-4, IL-10 and INFγ of ileal, colonic and MLN tissues were performed, as well as the immunohistochemical characterization of immune cells. The E. ninakohlyakimovae primary infection resulted in moderate to severe enteritis with different degrees of diarrhoea and was accompanied by high OPG counts and an increase of most immune cells analyzed when compared to uninfected control animals. Furthermore, eosinophil-, lymphocyte-, globular leukocyte- and mast cell-counts were significantly higher in the challenge group compared to the primary infected animals, whilst the opposite was true for PMN counts. The challenge infection was also associated with moderate increased levels of local mucosal IgA. Interestingly, the number of immature schizonts found at the ileal mucosa was statistically higher in the challenge infected

  16. Mosquito abundance, bed net coverage and other factors associated with variations in sporozoite infectivity rates in four villages of rural Tanzania

    DEFF Research Database (Denmark)

    Kweka, Eliningaya J; Nkya, Watoky M M; Mahande, Aneth M

    2008-01-01

    . Sporozoite infectivity rates, mosquito host blood meal source, bed net coverage and mosquito abundance were assessed in this study. METHODOLOGY: A longitudinal survey was conducted in four villages in two regions of Tanzania. Malaria vectors were sampled using the CDC light trap and pyrethrum spray catch...... methods. In each village, ten paired houses were selected for mosquitoes sampling. Sampling was done in fortnight case and study was undertaken for six months in both Kilimanjaro (Northern Tanzania) and Dodoma (Central Tanzania) regions. RESULTS: A total of 6,883 mosquitoes were collected including: 5......,628 (81.8%) Anopheles arabiensis, 1,100 (15.9%) Culex quinquefasciatus, 89 (1.4%) Anopheles funestus, and 66 (0.9%) Anopheles gambiae s.s. Of the total mosquitoes collected 3,861 were captured by CDC light trap and 3,022 by the pyrethrum spray catch method. The overall light trap: spray catch ratio was 1...

  17. Coccidiosis due to various species of Eimeria in the stunted and diarrheic native turkey poults: Pathology and morphological characterization of oocysts

    Directory of Open Access Journals (Sweden)

    Dezfoulian, O.

    2010-01-01

    Full Text Available Fecal samples of 60 turkey poults that showed chronic progressive symptoms like unthriftiness, loss ofweight, diarrhea were collected from the most rural areas with high rate of turkey population in north andwest part of country for intestinal protozoan parasites. According to the morphological characteristics, likeshape, presence or absence of micropyle, and/or polar granule, the 5 different types of eimerian oocycts were diagnosed in the stool of infected birds, including E. adenoids, E. meleagridis, E. dispersa, Eimeria spp (E. innocua or E. subrotunda and E. meleagrimitis. Various life- cycle stages of Eimeria were identified in the epithelial lining of inflamed intestine of the affected turkey poults.

  18. Expression of nutrient transporters in duodenum, jejunum, and ileum of Eimeria maxima-infected broiler chickens.

    Science.gov (United States)

    Fetterer, Raymond H; Miska, Katarzyna B; Jenkins, Mark C; Wong, Eric A

    2014-10-01

    The uptake of amino acids is mediated by active transporters located on the basolateral and brush border membranes of intestinal epithelial cells. The current study investigated the expression of amino acid transporters (AAT) and other genes in the intestine of chicks infected with Eimeria maxima. At 7-day postinfection (PI), tissue from each intestinal segment (duodenum, jejunum, and ileum) was taken from birds inoculated with 3 × 10(3) oocysts/bird and processed to recover RNA. Analysis of gene expression was performed using real-time reverse transcription polymerase chain reaction (qRT-PCR). Results were given as relative expression using β₂-microglobulin as an endogenous control. All the genes studied were expressed in three segments of the intestines, and expression of the genes was altered by infection with E. maxima. Even though the jejunum is considered the parasite's primary predilection site, there was no segment-related difference in expression of most of the genes studied. The antimicrobial peptide (LEAP2) was downregulated in all three segments of the intestine. The results also demonstrate that transporters associated with brush border membranes were downregulated while transporters associated with the basolateral membranes were upregulated and that E. maxima alters the expression of AAT and LEAP2 throughout the small intestine.

  19. Immune protection of microneme 7 (EmMIC7) against Eimeria maxima challenge in chickens.

    Science.gov (United States)

    Huang, Jingwei; Zhang, Zhenchao; Li, Menghui; Song, Xiaokai; Yan, Ruofeng; Xu, Lixin; Li, Xiangrui

    2015-10-01

    In the present study, the immune protective effects of recombinant microneme protein 7 of Eimeria maxima (rEmMIC7) and a DNA vaccine encoding this antigen (pVAX1-EmMIC7) on experimental challenge were evaluated. Two-week-old chickens were randomly divided into five groups. Experimental groups of chickens were immunized with 100 μg DNA vaccine pVAX1-MIC7 or 200 μg rEmMIC7, while control groups of chickens were injected with pVAX1 plasmid or sterile phosphate buffered saline (PBS). The results showed that the anti-EmMIC7 antibody titres in chickens of both rEmMIC7 and pVAX1-MIC7 groups were significantly higher as compared to PBS and pVAX1 control (P maxima challenge in chickens and it could be an effective antigen candidate for the development of new vaccines against E. maxima.

  20. Course of induced infection by Eimeria krijgsmannni in immunocompetent and immunodeficient mice.

    Science.gov (United States)

    Ono, Yuina; Matsubayashi, Makoto; Kawaguchi, Hiroaki; Tsujio, Masashi; Mizuno, Masanobu; Tanaka, Tetsuya; Masatani, Tatsunori; Matsui, Toshihiro; Matsuo, Tomohide

    2016-01-01

    Recently, we have demonstrated the utility of Eimeria krijgsmanni as a novel mouse eimerian parasite for elucidating the biological diversity. The parasite showed notable infectivity to mice with various levels of immune status and susceptibility to antimicrobial agents including coccidiostat. However, the detailed lifecycle of E. krijgsmanni had not yet been determined and this information was lacking in discussion of previous findings. In the present study, we clarified the morphological characteristics of E. krijgsmanni and its lifecycle in normal mice, and examined the effects in immunodeficient mice and lifecycle stage for challenge infections after the primary inoculation. In immunocompetent mice, the lifecycle consisted of four asexual stages and the sexual sages followed by formation of oocysts during the prepatent periods. Interestingly, the second-generation meronts were detected in all observation periods after the disappearance of the other stages. For the challenge infection of immunodeficient mice, all developmental stages except for the second generation meronts were temporarily vanished. This finding suggests a "rest" or marked delay in development and a "restart" of the promotion toward the next generations. The second generation meronts may play an important role in the lifecycle of E. krijgsmanni.

  1. Epidemiology of Eimeria infections in sheep raised extensively in a semiarid region of Brazil

    Directory of Open Access Journals (Sweden)

    Luiz Eduardo Barreto de Souza

    Full Text Available Abstract The aim of this study was to identify and determine the prevalence of Eimeria species affecting sheep raised extensively in a semiarid region of Brazil. Fecal samples of native sheep were collected during the rainy and dry seasons. The degree of infection was determined by counting oocysts per gram (OPG of feces, and the morphometric method was used for species identification. Oocysts were found in all the properties assessed, in which 68.3% of the animals were infected. The prevalence of oocysts was influenced by the season and animal category (P<0.05. It was higher during the rainy season than the dry season (80.2% vs. 55.8% and highest in young animals than the adults animals (68.2% vs. 39.6%. The OPG was lower during the dry season (1,269 ± 312 vs. 4,400 ± 1,122. Ten species were found; of these, E. ovinoidalis, E. granulosa, E. faurei, and E. crandallis were the most frequent. E. ovinoidalis and E. crandallis were found in all properties, with their prevalences being 19.4% and 13.6% respectively. The high prevalence of pathogenic species shows that eimeriosis is a risk for animals raised extensively in the semiarid region.

  2. The endogenous development and pathogenicity of Eimeria anseris (Kotlan, 1932) in domestic goslings.

    Science.gov (United States)

    Song, Hongqin; Liu, Dandan; Xu, Jinjun; Wu, Lili; Dai, Yabin; Liu, Mei; Tao, Jianping

    2017-01-01

    Twenty-one, 25-day-old, artificially reared, coccidia-free goslings (Anser cygnoides var. domestica) were inoculated orally with 0.5 × 10 4 , 1 × 10 4 , or 100 × 10 4 sporulated oocysts of Eimeria anseris and sacrificed at intervals from 24 to 216 h post-inoculation (HPI). Nine uninfected goslings served as negative controls. Parts of the visceral organs from goslings, including the intestines, kidneys, and liver, were fixed, sectioned, and observed microscopically. The results revealed that two generations of meronts occurred in the life cycle of E. anseris. The first generation of meronts developed at 24-96 HPI and the second generation at 90-128 HPI. Each meront contained 4-10 merozoites. Development of gamonts began at 128 HPI and mature oocysts appeared at 168 HPI. Developmental stages presented mainly in the epithelial cells of crypts and lamina propria in the posterior parts of the jejunum and ileum. Parasites localized mostly in the cytoplasm and occasionally in the nuclei of host cells. Histological lesions were pronounced in the jejunum and ileum. Desquamation and necrosis of the epithelium of intestine and crypts, infiltration of inflammatory cells, and hemorrhage and mucosal edema were associated with aggregates of endogenous stages. The infected goslings mainly showed severe diarrhea, depression, anorexia, and emaciation, suggesting that E. anseris is highly pathogenic in goslings.

  3. [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.

  4. Regeneration of the intestinal mucosa in Eimeria and E. Coli challenged broilers supplemented with amino acids.

    Science.gov (United States)

    Gottardo, E T; Prokoski, K; Horn, D; Viott, A D; Santos, T C; Fernandes, J I M

    2016-05-01

    The aim of this study was to evaluate the regeneration of the intestinal mucosa in Eimeria and E. coli challenged broilers supplemented with glutamine, arginine, and threonine. Six hundred male broilers at one d of age from the Cobb strain were utilized. The design was completely randomized using a 2×3 factorial design (unchallenged and challenged and 3 diets). A commercial diet was used as a control and 2 other diets were formulated with glutamine (1.5 and 3% Aminogut®), arginine (1 and 2% L-Arginine), and threonine (1 and 2% L-threonine). The animals that consumed diets supplemented with amino acids presented better (Pbroilers that received diets supplemented with amino acids. High levels of amino acids in the experimental feeds reflected in greater protein levels in poultry house litter, and they did not interfere with ammonia production. The supplementation of diets with trophic amino acids can positively contribute to the regeneration and proliferation of the intestinal mucosa in broilers and to the maintenance of zootechnical performance when submitted to enteric challenges. © 2016 Poultry Science Association Inc.

  5. Protective effects of Aloe vera-based diets in Eimeria maxima-infected broiler chickens.

    Science.gov (United States)

    Yim, Dongjean; Kang, Sang S; Kim, Dong W; Kim, Sang H; Lillehoj, Hyun S; Min, Wongi

    2011-01-01

    Aloes have been widely used for a broad range of pharmacological activities, including parasitic problems. Avian coccidiosis is the most costly and wide-spread parasitic disease in the poultry industry, and has been mainly controlled by the use of chemotherapeutic agents. Due to the emergence of drug-resistant strains, alternative control strategies are needed. In this study, the protective effects of Aloe vera-based diets were assessed in broiler chickens following oral infection with Eimeria maxima. Chickens were fed a regular diet supplemented with ground Aloe vera throughout the duration of the experiment beginning 2 days prior to infection with 1 × 10(4) sporulated oocysts of E. maxima. No significant differences were found in body weight gain or loss between the Aloe vera-supplemented and unsupplemented groups with or without E. maxima infections. Fecal oocyst shedding decreased significantly (p vera as compared to the unsupplemented group. Furthermore, the Aloe vera-supplemented group showed significantly fewer intestinal lesions (p vera could be used an alternative treatment for controlling avian coccidiosis. Copyright © 2010 Elsevier Inc. All rights reserved.

  6. Evaluation of pomegranate (Punica granatum pericarp aqueous extract on Eimeria spp. from Japanese quails (Coturnix japonica

    Directory of Open Access Journals (Sweden)

    Bruno P. Berto

    2014-06-01

    Full Text Available Antibacterial, anti-inflammatory and antiparasitic properties have been associated with the extract of pomegranate (Punica granatum in several animals and conditions. 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 study investigated the effects of the pomegranate pericarp aqueous extract on the shedding, viability and morphometry of three Eimeria spp. from Japanese quails, besides the weight gain and genotoxic activity. Although the pomegranate is recognized by multiple properties, including anti-coccidial, in the current study the results are contrary. The treated group shed greater amount of oocysts; the sporulation times and viability were similar in both groups; despite some morphometric differences, these were not expressive; weight gains were similar; and the pomegranate had insignificant effect genotoxic. Finally, these results suggest that the pomegranate pericarp extract did not influence on Eimeira spp. from Japanese quails; therefore, the pomegranate pericarp extract is not suggested in the prevention/treatment of coccidiosis in Japanese quails, or at least not using methods of preparation and administration applied in this study.

  7. Efficacy and economic analysis of two treatment regimens using toltrazuril in lambs naturally infected with Eimeria spp. on pasture.

    Science.gov (United States)

    de Souza Rodrigues, Fernando; Cezar, Alfredo Skrebsky; de Menezes, Fernanda Rezer; Sangioni, Luis Antônio; Vogel, Fernanda Silveira Flores; de Avila Botton, Sônia

    2017-11-01

    This study evaluated the efficacy and the economic viability of two anticoccidial treatment regimens tested in lambs naturally exposed to Eimeria spp. re-infections in a grazing system during a 140-day period. Twenty-four suckling lambs were distributed into three groups based on the individual count of oocysts per gram of feces (OPG) and body weight. Animals were treated with toltrazuril 5% (20 mg/kg) at 14- (GI) or 21-day (GII) intervals, and GIII was kept as untreated control. A cost-benefit analysis of each treatment regimen was calculated. Additionally, economic analysis was performed on four hypothetical scenarios, in which lambs could be having 10, 25, 50, or 85% decrease in their expected body weight gain due to clinical. Efficacy of toltrazuril against Eimeria spp. was 96.9-99.9% (GI) and 74.2-99.9% (GII). E. ovinoidalis was most frequently identified, but no clinical signs of coccidiosis were observed in lambs. There were no differences in weight gain among the groups. The cost of treatment per lamb was $13.09 (GI) and $7.83 (GII). The estimation model showed that the cost-benefit ratio favored treatment with toltrazuril when lambs fail to gain weight. In the studied flock, the break-even point for toltrazuril administered at 14-day intervals was reached with 85% decrease in mean weight gain. In conclusion, toltrazuril can be used at 14-day intervals to control Eimeria spp. (re)-infection in lambs raised on pasture. This treatment regimen was not economically feasible for subclinical coccidiosis; however, it may be feasible when used to prevent weight loss caused by clinical coccidiosis.

  8. Temporal pattern changes in duodenal protein tyrosine nitration events in response to Eimeria acervulina infection in chickens.

    Science.gov (United States)

    Elsasser, Ted H; Miska, Kate; Kahl, Stanislaw; Fetterer, Raymond H; Martínez Ramirez, Alfredo

    2018-06-04

    Intracellular generation of nitric oxide (NO) and superoxide anion (SOA) can result in the formation of 3'-nitrotyrosine proteins (NTp). Nitrated proteins usually are associated with significant perturbation in protein function, apoptosis, autophagy, and cell death. We undertook the present study to establish the temporal dynamics of NTp generation in cytokeratin-18-positive epithelial cells (ETCs) of broiler chickens in response to infection with Eimeria acervulina. Duodenal tissue was harvested from noninfected (NOI) and infected (INF) broilers on days (d) 1, 3, 6, 7, and 10 postinfection (PI) and fixed, embedded, and sectioned for quantitative image analysis, immunohistochemistry with antibodies specific to NTp and the SOA-generating enzyme xanthine oxidase (XO). The pixel density characteristics for NTp and XO representative of ETCs demonstrated that NTp and XO increased in intestinal villi as early as d1 PI (P ETCs through d6 PI. For XO, increases in cell content increased only through d3. On d6 and d7 PI, high levels of NTp were present in immune infiltrating cells (IIC) where no XO was detected. The increases in ETC NTp occurred in a defined pattern, significant by villus-to-crypt location for day of infection, initiating in the distal villus and progressing down into the crypts. Two NTp patterns were observed for ETCs: a high level associated with ETCs harboring parasites and a low-level increase in ETCs not containing Eimeria but in proximity to such. The data suggest that NTp and XO responses may mediate some of the processes through which ETCs respond to Eimeria to limit the extent of infection by this pathogen.

  9. Effects of methylglyoxal bis(guanylhydrazone) and two phenylated analogues on S-adenosylmethionine decarboxylase activity from Eimeria stiedai (Apicomplexa).

    Science.gov (United States)

    San-Martín Núñez, B; Alunda, J M; Balaña-Fouce, R; Ordóñez Escudero, D

    1987-01-01

    1. Activity of S-adenosylmethionine decarboxylase, one of the rate-limiting enzymes of polyamine biosynthesis, was determined in oocysts of Eimeria stiedai, a coccidian parasite of the rabbit. 2. Several properties of the enzyme were compared to the mammalian enzyme. It showed considerably less substrate affinity than the analog enzyme from the rabbit. 3. The E. stiedai enzyme showed a low sensitivity to methylglyoxal bis(guanylhydrazone), a frequently used inhibitor of the enzyme in mammals, and two phenylated derivatives. 4. Results with the inhibitors are discussed in view of their potential use in chemotherapy.

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

    International Nuclear Information System (INIS)

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

    1977-01-01

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

  11. Resistance to infection with Eimeria vermiformis in mouse radiation chimeras is determined by donor bone-marrow cells

    International Nuclear Information System (INIS)

    Joysey, H.S.; Wakelin, D.; Rose, M.E.

    1988-01-01

    The course of infection with Eimeria vermiformis was determined in BALB/b, BALB/c, and C57BL/10ScSn (B10) mice and in radiation chimeras prepared from the H-2-compatible BALB/b and B10 mice. The BALB strains, irrespective of H-2 haplotype, were resistant, the B10 mice were susceptible, and in the chimeras infection was characterized by the genotype of the donated bone-marrow cells and not by the phenotype of the recipient. Thus, the genetic control of relative resistance or susceptibility to infection with this parasite is expressed through bone-marrow-derived cells

  12. Description of Eimeria arabukosokokensis sp. n. (Apicomplexa: Eimeriidae) from Telescopus semiannulatus (Serpentes: Colubridae) with notes on eimerian coccidia from snakes of Eastern Kenya

    Czech Academy of Sciences Publication Activity Database

    Šlapeta, Jan Roger; Modrý, David; Ashe, J.; Koudela, Břetislav

    2003-01-01

    Roč. 50, č. 1 (2003), s. 23-30 ISSN 0015-5683 R&D Projects: GA ČR GA524/00/P015 Institutional research plan: CEZ:AV0Z6022909 Keywords : Coccidia * Caryospora * Eimeria Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 0.469, year: 2003

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

    DEFF Research Database (Denmark)

    Almeida, Gustavo Fonseca; Thamsborg, Stig Milan; Madeira, Alda M.B.N

    2014-01-01

    and oocyst excretion were investigated. Broilers given chemical coccidiostats performed better than all other groups. Broilers given the two highest dosages of the herbal mixture had intermediate lesion scores caused by Eimeria acervulina, which was higher than in broilers given coccidiostats, but less than...

  14. New data on Eimeria dicentrarchi (Apicomplexa: Eimeriidae), a common parasite of farmed European sea bass (Dicentrarchus labrax) from the mid-eastern Adriatic

    Science.gov (United States)

    This study extends the original description of Eimeria dicentrarchi Daoudi and Marquès, 1987, a common coccidian parasite of European sea bass from the mid-eastern Adriatic, by providing insights into the parasite’s site of infection, development and pathogenicity. E. dicentrarchi was found in vario...

  15. Eimeria-induced chicken cNK-2 is an anti-infective host defense peptide and an immunomodulator of host innate immunity

    Science.gov (United States)

    Avian coccidiosis is one of the most widespread infectious diseases of chickens. The etiologic agent of avian coccidiosis is Eimeria, a genus of eukaryotic obligate intracellular parasites belonging to the phylum Apicomplexa. Clinical manifestations of infection include damage to the intestinal epit...

  16. Synergistic effect of embryo vaccination with Eimeria profilin and Clostridium perfringens NetB proteins on inducing protective immunity against necrotic enteritis in broiler chickens

    Science.gov (United States)

    The effects of embryo vaccination with Eimeria profilin plus Clostridium perfringens NetB toxin proteins in combination with the Montanide IMS-OVO adjuvant on the chicken immune response to necrotic enteritis were investigated using an E. maxima/C. perfringens co-infection model. Eighteen-day-old br...

  17. Two Eimerian Coccidia (Apicomplexa : Eimeriidae) from the critically endangered Arakan forest turtle Heosemys depressa (Testudines: Geoemydidae), with description of Eimeria arakanensis n. sp.

    Czech Academy of Sciences Publication Activity Database

    Široký, P.; Modrý, David

    2006-01-01

    Roč. 45, č. 2 (2006), s. 183-189 ISSN 0065-1583 R&D Projects: GA ČR GP524/03/D104 Institutional research plan: CEZ:AV0Z60220518 Keywords : Eimeria * Apicomplexa * Heosemys Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 1.162, year: 2006

  18. Eimeria atheridis n. sp. (Apicomplexa: Eimeriidae), a new coccidium from the western bush viper Atheris chlorechis (Pel, 1851) from tropical Africa

    Czech Academy of Sciences Publication Activity Database

    Modrý, David; Nečas, T.; Mazuch, T.; Kamler, M.

    2004-01-01

    Roč. 59, č. 1 (2004), s. 71-74 ISSN 0165-5752 R&D Projects: GA ČR GP524/03/D104; GA ČR GD524/03/H133 Institutional research plan: CEZ:AV0Z6022909 Keywords : Apicomplexa * Eimeriidae * Eimeria Subject RIV: EG - Zoology Impact factor: 0.669, year: 2004

  19. Efficacy of a DNA Vaccine Carrying Eimeria maxima Gam56 Antigen Gene against Coccidiosis in Chickens

    Science.gov (United States)

    Xu, Jinjun; Zhang, Yan

    2013-01-01

    To control coccidiosis without using prophylactic medications, a DNA vaccine targeting the gametophyte antigen Gam56 from Eimeria maxima in chickens was constructed, and the immunogenicity and protective effects were evaluated. The ORF of Gam56 gene was cloned into an eukaryotic expression vector pcDNA3.1(zeo)+. Expression of Gam56 protein in COS-7 cells transfected with recombinant plasmid pcDNA-Gam56 was confirmed by indirect immunofluorescence assay. The DNA vaccine was injected intramuscularly to yellow feathered broilers of 1-week old at 3 dosages (25, 50, and 100 µg/chick). Injection was repeated once 1 week later. One week after the second injection, birds were challenged orally with 5×104 sporulated oocysts of E. maxima, then weighed and killed at day 8 post challenge. Blood samples were collected and examined for specific peripheral blood lymphocyte proliferation activity and serum antibody levels. Compared with control groups, the administration of pcDNA-Gam56 vaccine markedly increased the lymphocyte proliferation activity (P<0.05) at day 7 and 14 after the first immunization. The level of lymphocyte proliferation started to decrease on day 21 after the first immunization. A similar trend was seen in specific antibody levels. Among the 3 pcDNA-Gam56 immunized groups, the median dosage group displayed the highest lymphocyte proliferation and antibody levels (P<0.05). The median dosage group had the greatest relative body weight gain (89.7%), and the greatest oocyst shedding reduction (53.7%). These results indicate that median dosage of DNA vaccine had good immunogenicity and immune protection effects, and may be used in field applications for coccidiosis control. PMID:23710081

  20. Molecular diagnosis of Eimeria stiedae in hepatic tissue of experimentally infected rabbits.

    Science.gov (United States)

    Hassan, Khaled M; Arafa, Waleed M; Mousa, Waheed M; Shokier, Khaled A M; Shany, Salama A; Aboelhadid, Shawky M

    2016-10-01

    The early detection of Eimeria stiedae in the hepatic tissue of experimentally infected rabbits was investigated using molecular assay. Forty 6-week-old male New Zealand rabbits were divided into two groups. Group A (30 animals) was infected with 2.5 × 10(4) sporulated oocysts of E. stiedae per animal on Day 0 and Group B (10 animals) was used as the uninfected controls. Three animals from Group A and one from Group B were sacrificed at 0, 3, 6, 9, 12, 15, 18, 21, 24 and 27 days post infection (PI). Gross and microscopic post-mortem findings were recorded. Polymerase chain reaction (PCR) of the E. stiedae internal transcribed spacer 1 genomic region was conducted on blood, liver tissue, and feces from the Group A experimentally infected animals. Macroscopically, the liver showed irregular yellowish white nodules pathognomonic to E. stiedae infection beginning on Day 15 PI. Hepatomegaly and ascites were obvious from Day 21-24 PI. The presence of different E. stiedae schizonts and gametocytes in the histopathological sections of the biliary epithelium were evident on Day 15 PI. The E. stiedae PCR was first positive in liver tissues on Day 12 and in fecal samples on Day 18 PI, but the blood samples were negative. In conclusion, the PCR can be used for early diagnosis and control of E. stiedae schizonts before shedding of the oocysts in feces. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Histomorphometry and macroscopic intestinal lesions in broilers infected with Eimeria acervulina.

    Science.gov (United States)

    Assis, R C L; Luns, F D; Beletti, M E; Assis, R L; Nasser, N M; Faria, E S M; Cury, M C

    2010-03-25

    This study aimed at measuring intestinal villi and assessing the intestinal absorptive area in broilers infected with Eimeria acervulina under different treatments to control coccidiosis. The experiment was divided into two stages, carried out in successive housings, raised in the same environment (or aviary). In the first stage, on 25 May 2008, fifty 12-day-old birds were orally inoculated with 3 x 10(3) oocysts of E. acervulina. In the second stage, on July 2008, other 50 birds were allocated on litter contaminated by the feces of birds on the first housing (natural infection by oocysts present in the reused litter). The experiment was arranged in a complete randomized design with five treatments and three replicates of 10 chicks per treatment. Broiler chicks were housed at 1 day of age and autopsies were performed at 21 days of age. Three 2-cm-long segments of the duodenum were excised from each bird and fixed in 10% buffered formalin. A total of 30 slides were prepared for each treatment, totaling 150 evaluated histological sections using H&E staining. Villus morphology was carried out by the HL Image 97 software. The intestinal absorptive area was calculated and macroscopic lesions were classified according to standard lesion scores. Results showed that intestinal villus measurements and absorptive area are directly affected by E. acervulina and that there is direct and positive correlation between the macro and microscopic findings observed in intestinal coccidiosis. E. acervulina causes shortening of villi and reduction in the intestinal absorptive area, affecting broiler growth. The prevention method of litter fermentation during the interval between housings and oral administration of Diclazuril can reduce the severity of intestinal lesions by E. acervulina in broilers impairing oocyst virulence or viability.

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

    Science.gov (United States)

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

    2014-01-01

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

  3. A new species of Eimeria Schneider, 1875 (Apicomplexa: Eimeriidae) from Alectoris barbara (Aves: Phasianidae) from the Canary Islands (Spain).

    Science.gov (United States)

    Fernández-Alvarez, A; Modry, D; Foronda, P

    2016-05-01

    The present study was conducted with the objective of identifying the species of Eimeria present in a cynegetic farm. A new coccidian (Apicomplexa: Eimeriidae) species is described from Barbary partridge, Alectoris barbara, from the Canary Islands. Experimental infections were carried out in order to determine the prepatent period, sporulation time, site of infection, and morphology of endogenous stages. One species is described as new. Eimeria barbarae n. sp. has ellipsoidal oocysts, 20.0 × 14.4 (16-23 × 13-16) μm, with a shape-index (SI) of 1.39. Sporocysts are almond-shaped, 9.0 × 5.4 (6.5-11 × 4.5-6) μm, SI = 1.56. The endogenous development takes place along the intestine. The present study showed that E. barbarae causes severe pathologies in A. barbara chickens, with impact on their health condition. Control strategies needs to be implemented to reduce the loss due to coccidiosis at studied farm.

  4. Co-localization of a CD1d-binding glycolipid with a radiation-attenuated sporozoite vaccine in LN-resident DCs for a robust adjuvant effect

    Science.gov (United States)

    Li, Xiangming; Kawamura, Akira; Andrews, Chasity D.; Miller, Jessica L.; Wu, Douglass; Tsao, Tiffany; Zhang, Min; Oren, Deena; Padte, Neal N.; Porcelli, Steven A.; Wong, Chi-Huey; Kappe, Stefan H. I.; Ho, David D.; Tsuji, Moriya

    2015-01-01

    A CD1d-binding glycolipid, α-Galactosylceramide (αGalCer), activates invariant natural killer T (iNKT) cells and acts as an adjuvant. We previously identified a fluorinated phenyl ring-modified αGalCer analog, 7DW8-5, displaying nearly 100-fold stronger CD1d binding affinity. In the present study, 7DW8-5 was found to exert a more potent adjuvant effect than αGalCer for a vaccine based on radiation-attenuated sporozoites (RAS) of a rodent malaria parasite, Plasmodium yoelii, also referred to as irradiated P. yoelii sporozoites (IrPySpz). 7DW8-5 had a superb adjuvant effect only when the glycolipid and IrPySpz were conjointly administered intramuscularly (i.m.). Therefore, we evaluated the impact of distinctly different biodistribution patterns of αGalCer and 7DW8-5 on their respective adjuvant activities. While both glycolipids induce a similar cytokine response in sera of mice injected intravenously, after i.m. injection, αGalCer induces a systemic cytokine response, whereas 7DW8-5 is locally trapped by CD1d expressed by dendritic cells (DCs) in draining lymph nodes (dLNs). Moreover, the i.m. co-administration of 7DW8-5 with IrPySpz results in the recruitment of DCs to dLNs and the activation and maturation of DCs. These events cause the potent adjuvant effect of 7DW8-5, resulting in the enhancement of the CD8+ T-cell response induced by IrPySpz, and, ultimately, improved protection against malaria. Our study is the first to show that the co-localization of a CD1d-binding iNKT-cell stimulatory glycolipid and a vaccine, like RAS, in dLN-resident DCs upon i.m. conjoin administration governs the potency of the adjuvant effect of the glycolipid. PMID:26254338

  5. Differences in fecundity of Eimeria maxima strains exhibiting different levels of pathogenicity in its avian host.

    Science.gov (United States)

    Jenkins, Mark C; Dubey, J P; Miska, Katarzyna; Fetterer, Raymond

    2017-03-15

    Eimeria maxima is one of the most pathogenic species of avian coccidia, yet it is unknown why different E. maxima strains differ in the pathogenic effects they cause in chickens. The purpose of this study was to determine if a more pathogenic E. maxima strain (APU1) was also more fecund than a less pathogenic E. maxima strain (APU2). At identical doses, E. maxima APU1 always produces greater intestinal lesions and lower weight gain compared to E. maxima APU2. Using a dose response study, median and mean intestinal lesion scores in E. maxima APU1-infected chickens were greater by a score of 1-1.5 compared to chickens infected with E. maxima APU2. Likewise, weight gain depression in E. maxima APU1-infected chickens was 20-25% greater (equivalent to 110-130g body weight) than in E. maxima APU2-infected chickens. In order to understand the underlying cause of these observed clinical effects, 120 broiler chicks (5 oocyst levels, 6 replicates/level) were inoculated with various doses of E. maxima APU1 or APU2 oocysts. The dynamics of oocyst shedding was investigated by collecting fecal material every 12h from 114 to 210h post-inoculation (p.i.) and every 24h thereafter from 210 to 306h, and then processed for measuring E. maxima oocyst output. Oocysts were first observed at 138h p.i., and time of peak oocyst production was nearly identical for both E. maxima APU1 and APU2 around 150-162h. Total oocyst production was 1.1-2.6 fold higher at all dose levels for E. maxima APU1 compared to E. maxima APU2, being significantly higher (P<0.05) at the log 1.5 dose level. Other groups of chickens were infected with higher doses of E. maxima APU1 or APU2 oocysts, and intestinal lesions were assessed by histology at 72, 96, 120, and 144h p.i. Although schizonts, gamonts, and oocysts were observed at expected time-points, no obvious differences were noted in lesions induced by the two E. maxima strains. This study showed that the greater fecundity of E. maxima APU1 compared to E

  6. Anticoccidial efficacy of naringenin and a grapefruit peel extract in growing lambs naturally-infected with Eimeria spp.

    Science.gov (United States)

    Pérez-Fonseca, Agustín; Alcala-Canto, Yazmin; Salem, Abdelfattah Z M; Alberti-Navarro, Aldo B

    2016-12-15

    The current study aimed to determine the anti-Eimeria efficacy of an extract of grapefruit peels (GF) and commercial naringenin (NAR) in naturally-infected lambs, as well as the influence of these flavonoids on the oxidative status during ovine coccidiosis. Pharmacokinetic profiles were also determined. Extracts were administered per os to Eimeria naturally infected growing lambs during 90 consecutive days. The commercial anticoccidial drug toltrazuril (TTZ) was included in this trial as a standard. Twenty-four lambs were divided into four groups: NAR, lambs given a daily dose of 5mg of a commercial naringenin extract of 98% higher purity per kg body weight; GF, lambs that recived a daily dose of 5mg of ethanolic extract of grapefruit peels per kg body weight; TTZ, lambs treated with 20mg of toltrazuril/kg body weight on days 0 and 15 of the experiment; and CTRL, untreated lambs that received daily dose of 30ml of water. Daily doses of GF and NAR were dissolved in 30ml of water and orally given to animals; whereas toltrazuril was administered as a single dose of an undiluted suspension to lambs of the TTZ group. The CTRL group received 30ml of water; as well as the TTZ group for the period after the single dose administration. Fecal and serum samples were collected from all lambs. Anticoccidial efficacy was estimated by coprological techniques. Generation of nitric oxide levels and the antioxidant capacity of the experimental compounds were determined by the Griess and ABTS assays, respectively. The pharmacokinetic parameters of NAR and the GF extract were obtained. On day 30 post-ingestion, anticoccidial efficacy was 91.76% (NAR) and 89.65% (GF); whereas 99.63% of efficacy was achieved with TTZ 15days after treatment. NAR, GF and TTZ significantly reduced oxidative stress in infected animals. The mean daily weight gain for each group was 122g (NAR), 122g (GF), 143g (TTZ) and 98g (CTRL). Following the oral administration of NAR and GF, values in plasma approached

  7. Lipid and glucose metabolism of broilers (Gallus gallus domesticus experimentally infected with Eimeria acervulina Tyzzer, 1929 oocysts

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    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. Electron microscopic study on macrogametogenesis of Eimeria labbeana infecting the Egyptian wild doves and host-parasite relationship.

    Science.gov (United States)

    Bashtar, A R; Abdel-Ghaffar, F A; Ahmed, A K

    1991-04-01

    The development of macrogametes of Eimeria labbeana was studied by electron microscopy in the epithelial cells of the villi at 96 hrs. post-infection. Appearance of young macrogamonts was characterized by the loss of the architecture of the apicomplexa (polar ring, rhoptries, micronemes, conoid, subpellicular microtubules), while the pellicle became only one unit membrane. This was associated by the formation of wall forming bodies II then I. Moreover, the mitochondria, endoplasmic reticulum and Golgi were increased in the cytoplasm. Amylopectin granules as well as lipid globules were greatly increased in mature macrogametes. Host cell reaction due to infection included enlargement and deformation of infected cells, hypertrophy of their nuclei, swollen and degeneration of mitochondria, endoplasmic reticulum and vacuolation of ground cytoplasm. These changes occur in both cells with and without parasite.

  9. Eimeria pileata n. sp. (Apicomplexa: Eimeriidae) from the rufous-capped brush finch Atlapetes pileatus Wagler (Passeriformes: Emberizidae) in Mexico.

    Science.gov (United States)

    Soriano-Vargas, Edgardo; Medina, Juan Pablo; Salgado-Miranda, Celene; García-Conejo, Michele; Galindo-Sánchez, Karla Patrícia; Janczur, Mariusz Krzysztof; Berto, Bruno Pereira; Lopes, Carlos Wilson Gomes

    2015-11-01

    A new coccidian species (Protista: Apicomplexa: Eimeriidae) collected from the rufous-capped brush finch Atlapetes pileatus Wagler in the Nevado de Toluca Natural Protected Area, Mexico. Oöcysts of Eimeria pileata n. sp. are ellipsoidal, measuring on average 16.5 × 14.1 μm, with a smooth, bi-layered wall. Micropyle and oöcyst residuum are absent, but a polar granule is present. Sporocysts are ellipsoidal, measuring on average 9.0 × 5.4 μm. Stieda and sub-Stieda bodies are both present. A sporocyst residuum is present as a compact mass of granules. This is the third description of an eimeriid coccidian infecting passerines.

  10. Endogenous Life Cycle of Eimeria melanomytis (Apicomplexa: Eimeriidae) from the Dusky Rice Rat, Melanomys caliginosus (Rodentia: Cricetidae: Sigmodontinae) in Costa Rica.

    Science.gov (United States)

    Chinchilla, Misael; Valerio, Idalia; Sánchez, Ronald; Duszynski, Donald W

    2017-02-01

    Endogenous stages of the life cycle of Eimeria melanomytis, infecting the peripheral epithelial cells of villi of the small intestine of experimentally infected young dusky rice rats, Melanomys caliginosus , were studied. Giemsa-stained mucosal scrapings and histological sections were examined for all the stages. Eimeria melanomytis has 3 generations of meronts (M), different in size, shape, and number of merozoites (m); and in size, shape, and location of the nuclei within the cytoplasm of the meronts. The 3 meront types, M 1 -M 3 , respectively, had 11-14 (m 1 ), 7-10 (m 2 ), and 20-30 (m 3 ) merozoites. Macrogametocytes and microgametocytes, as well as macrogametes and microgametes, complete the sexual cycle forming the unsporulated oocysts. This parasite's endogenous development produced severe intestinal lesions in experimentally infected dusky rice rats.

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

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    Daszak P

    1998-01-01

    Full Text Available Three new species of Eimeria are described from iguanid lizards of Central and South America. The oocysts of each species have no micropyles or residua and the sporocysts lack Stieda bodies, but all have a sporocyst residuum. Eimeria sanctaluciae n.sp. was found in the St. Lucia tree lizard, Anolis luciae, collected from the Maria Islands, Lesser Antilles. The oocysts are spherical to subspherical, averaging 17.3 x 16.5 µm, with a single layered colourless wall; about 60% contain polar granules. The sporocysts are ellipsoidal and average 7.7 x 5.5 µm. Eimeria liolaemi n.sp. was recovered from the blue-gold swift, Liolaemus taenius, from Chile. The oocysts are spherical to subspherical, measuring 21 x 20.1 µm with a single-layered colourless wall. The sporocysts are subspherical and average 7.4 x 6.8 µm. Eimeria caesicia n.sp. is described from the Brazilian collared iguanid, Tropidurus torquatus. The oocysts measure 27.4 x 23.7 µm, are spherical to subspherical, with a bilayered wall, the outer surface of which appears pale blue in colour, the thin, inner wall appearing brown, when viewed by direct light under the optical microscope. The sporocysts are subspherical and average 9.4 x 7.2 µm. Unnamed polysporocystid oocysts with dizoic sporocysts are reported from the faeces of the lesser St. Vincent tree lizard, Anolis trinitatis and the possibility of spurious parasitism briefly discussed. In addition, oocysts of an unnamed Isospora sp. with a smooth oocyst wall which closely resembles I. reui were recovered from A. trinitatis.

  12. Eimeria species in young and adult sheep raised under intensive and / or semi-intensive systems of a herd from Umuarama city, Parana State, Brazil

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    Welber Daniel Zanetti Lopes

    2013-11-01

    Full Text Available The present study aimed to identify Eimeria species in young and adult sheep raised under intensive and / or semi-intensive systems of a herd from Umuarama city, Parana State, Brazil using the traditional diagnostic methods and to correlate the infection level/types of infection in the different age/system in this herd. Fecal samples were collected from the rectum of 210 sheep and were subjected to laboratory analysis to differentiate the species. Furthermore, animals were observed to determine the occurrences of the clinical or subclinical forms of eimeriosis. Out of the 210 collected fecal samples, 147 (70% were positive for Eimeria oocysts, and 101 (47.86% belonged to young animals that were raised under intensive and / or semi-intensive farming systems. Oocysts from 9 species of Eimeria parasites were identified in the sheep at the following prevalence rates: E. crandallis, 50.0%; E. parva, 21.6%; E. faurei, 8.1%; E. ahsata, 8.1%; E. intricata, 5.4%; E. granulosa , 2.7%; E. ovinoidalis , 2.0%; E. ovina , 1.3%; and E. bakuensis , 0.6%. There were no differences regarding the more frequent Eimeria species among the different ages of animals or between the different farming management systems. Based on these data, E. crandallis was the most prevalent, followed by E. parva and E. faurei species, regardless of the age. Higher parasitism was diagnosed in the young animals that were raised in a confinement regime, and the disease found in the herd was classified as subclinical. Further studies should be conducted in this herd, to verify if the eimeriosis subclinical can cause damage especially in young animals with a high level of infection.

  13. Evaluating Controlled Human Malaria Infection in Kenyan Adults with Varying Degrees of Prior Exposure to Plasmodium falciparum using sporozoites administered by intramuscular injection

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    Susanne Helena Hodgson

    2014-12-01

    Full Text Available Background: Controlled human malaria infection (CHMI studies are a vital tool to accelerate vaccine and drug development. As CHMI trials are performed in a controlled environment, they allow unprecedented, detailed evaluation of parasite growth dynamics (PGD and immunological responses. However, CHMI studies have not been routinely performed in malaria-endemic countries or used to investigate mechanisms of naturally-acquired immunity (NAI to Plasmodium falciparum. Methods: We conducted an open-label, randomized CHMI pilot-study using aseptic, cryopreserved P. falciparum sporozoites (PfSPZ Challenge to evaluate safety, infectivity and PGD in Kenyan adults with low to moderate prior exposure to P. falciparum (Pan African Clinical Trial Registry: PACTR20121100033272. Results: All participants developed blood-stage infection confirmed by qPCR. However one volunteer (110 remained asymptomatic and blood-film negative until day 21 post-injection of PfSPZ Challenge. This volunteer had a reduced parasite multiplication rate (PMR (1.3 in comparison to the other 27 volunteers (median 11.1. A significant correlation was seen between PMR and screening anti-schizont ELISA OD (p=0.044, R=-0.384 but not when volunteer 110 was excluded from the analysis (p=0.112, R=-0.313. Conclusions: PfSPZ Challenge is safe and infectious in malaria-endemic populations and could be used to assess the efficacy of malaria vaccines and drugs in African populations. Whilst our findings are limited by sample size, our pilot study has demonstrated for the first time that NAI may impact on PMR post-CHMI in a detectable fashion, an important finding that should be evaluated in further CHMI studies.

  14. Circumsporozoite Protein-Specific Kd-Restricted CD8+ T Cells Mediate Protective Antimalaria Immunity in Sporozoite-Immunized MHC-I-Kd Transgenic Mice

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    Jing Huang

    2014-01-01

    Full Text Available Although the roles of CD8+ T cells and a major preerythrocytic antigen, the circumsporozoite (CS protein, in contributing protective antimalaria immunity induced by radiation-attenuated sporozoites, have been shown by a number of studies, the extent to which these players contribute to antimalaria immunity is still unknown. To address this question, we have generated C57BL/6 (B6 transgenic (Tg mice, expressing Kd molecules under the MHC-I promoter, called MHC-I-Kd-Tg mice. In this study, we first determined that a single immunizing dose of IrPySpz induced a significant level of antimalaria protective immunity in MHC-I-Kd-Tg mice but not in B6 mice. Then, by depleting various T-cell subsets in vivo, we determined that CD8+ T cells are the main mediator of the protective immunity induced by IrPySpz. Furthermore, when we immunized (MHC-I-Kd-Tg × CS-Tg F1 mice with IrPySpz after crossing MHC-I-Kd-Tg mice with PyCS-transgenic mice (CS-Tg, which are unable to mount PyCS-specific immunity, we found that IrPySpz immunization failed to induce protective antimalaria immunity in (MHC-I-Kd-Tg × CS-Tg F1 mice, thus indicating the absence of PyCS antigen-dependent immunity in these mice. These results indicate that protective antimalaria immunity induced by IrPySpz in MHC-I-Kd-Tg mice is mediated by CS protein-specific, Kd-restricted CD8+ T cells.

  15. Coccidiosis: recent advancements in the immunobiology of Eimeria species, preventive measures, and the importance of vaccination as a control tool against these Apicomplexan parasites

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    Shivaramaiah C

    2014-04-01

    Full Text Available Chaitanya Shivaramaiah,1 John R Barta,2 Xochitl Hernandez-Velasco,3 Guillermo Téllez,1 Billy M Hargis11Department of Poultry Science, University of Arkansas, Fayetteville, AR, USA; 2Department of Pathobiology, University of Guelph, ON, Canada; 3Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autonoma de Mexico, MexicoAbstract: Coccidiosis, caused by parasites of the genus Eimeria, is probably the most expensive parasitic disease of poultry. Species of Eimeria are ubiquitous where poultry are raised and are known to cause drastic reductions in performance and induce mortality, thereby affecting the overall health status of poultry. Chemotherapy has been the predominant form of disease control for many years, even though vaccination is steadily gaining importance as a feasible control method. The objective of this review is to highlight recent advancements in understanding the role of host immunity against coccidiosis. In addition, pros and cons associated with chemotherapy and the role of vaccination as an increasingly popular disease control method are discussed. Finally, the role played by recombinant vaccines as a potential vaccination tool is highlighted. With interest growing rapidly in understanding host–parasite biology, recent developments in designing recombinant vaccines and potential epitopes that have shown promise are mentioned.Keywords: Eimeria, coccidiosis, chemotherapy, recombinant vaccines, immunity

  16. 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

  17. Risk factors associated with Cryptosporidia, Eimeria, and diarrhea in smallholder dairy farms in Mukurwe-ini Sub-County, Nyeri County, Kenya

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    S. G. Peter

    2016-08-01

    Full Text Available Aim: This study was undertaken to determine the household, calf management, and calf factors associated with the occurrence of Eimeria, Cryptosporidia, and diarrhea in pre-weaned calves reared in smallholder dairy farms in Mukurweini Sub-County of Nyeri County, Kenya. In addition, the study also evaluated factors associated with average daily weight gain in the same pre-weaned calves. Materials and Methods: A total of 112 newborn calves (63 males and 49 females on 111 farms (1 set of twins were followed for 2 months between June 2013 and August 2013. Two calves were lost to follow-up. A pre-tested questionnaire was used to collect data on household characteristics and calf management practices in the 111 selected farms. On the first visit to the farm (within 7 days of the birth of the calf, blood samples were collected from the jugular vein to assess the level of maternal immunity acquired by the calf, by determining the serum total protein and selenium concentration. At 4 and 6 weeks of age, fecal samples from the calves were collected to assess the presence of Cryptosporidia and Eimeria oocysts. Every 2 weeks for 2 months, the calves and their environments were examined, their 2-week consumption and health history were recorded, and weights were estimated with a weight tape. Each of the factors was evaluated in a univariable regression model and only those found to be significant (p≤0.20 were included in a multivariable model. Elimination of non-significant factors was done in the multivariable model through a backward elimination procedure so that only those variables which were confounders, and/or significant at (p≤0.05 remained in the final model. Results: About 37% (41/110 of the calves experienced diarrhea at least once during the 2-month study period. The overall period prevalence of Eimeria and Cryptosporidia was 42.7% (47/110 and 13.6% (15/110, respectively. Low serum protein was associated with 1.8 and 2.4 times the odds of

  18. 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

    infection by several organisms of medical and veterinary importance such as Eimeria, Plasmodium, Toxoplasma, Cyclospora, and Neospora could be developed. Here, we show that two tyrosine-rich precursor glycoproteins, gam56 and gam82, found in specialized organelles (wall-forming bodies) in the sexual stage...... (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......-mediated cross-linking might be an enzyme-catalyzed event. As such, the mechanism of oocyst wall formation in Eimeria, is analogous to the underlying mechanisms involved in the stabilization of extracellular matrices in a number of organisms, widely distributed in nature, including insect resilin, nematode...

  19. Avaliação da eficácia da lasalocida e de alguns fatores epidemiológicos de Eimeria spp. parasitando bezerros Nelore mantidos em regime de pastejo

    OpenAIRE

    Cruvinel, Leonardo Bueno; Borges, Dyego Gonçalves Lino; Nicaretta, João Eduardo; Bastos, Thiago Souza Azeredo; Moro, Elio; Gama, Rogério Dantas; Borges, Fernando de Almeida; Lopes, Welber Daniel Zanetti

    2017-01-01

    RESUMO: A principal importância da eimeriose em bovinos, se deve ao baixo desempenho produtivo que os animais demonstram quando esta enfermidade apresenta-se sob a forma sub-clínica. Como objetivos, o presente trabalho avaliou a eficácia do uso da lasalocida sódica contra espécies de Eimeria spp. parasitando bezerros; avaliou também o desempenho ponderal dos animais submetidos aos diferentes tratamentos e analisou alguns fatores epidemiológicos que possam interferir na infecção por Eimeria no...

  20. Antioxidant and Anti-Inflammatory Activities of Pomegranate (Punica granatum on Eimeria papillata-Induced Infection in Mice

    Directory of Open Access Journals (Sweden)

    Omar S. O. Amer

    2015-01-01

    Full Text Available Coccidiosis is the most prevalent disease causing widespread economic loss, especially in poultry farms. Here, we investigated the effects of pomegranate peel extract (PPE on the outcome of coccidiosis caused by Eimeria papillata in mice. The data showed that mice infected with E. papillata and treated with PPE revealed a significant decrease in the output of oocysts in their faeces by day 5 p.i. Infection also induced inflammation and injury of the jejunum. This was evidenced (i as increases in reactive oxygen species, (ii, as increased neutrophils and decreased lymphocytes in blood (ii as increased mRNA levels of inducible nitric oxide synthase (iNOS, Bcl-2 gene, and of the cytokines interferon gamma (IFN-γ, tumour necrosis factor-α (TNF-α, and interleukin-1β (IL-1β, and (iv as downregulation of mucin gene MUC2 mRNA. All these infection-induced parameters were significantly altered during PPE treatment. In particular, PPE counteracted the E. papillata-induced loss of the total antioxidant capacity. Our data indicated that PPE treatment significantly attenuated inflammation and injury of the jejunum induced by E. papillata infections.

  1. Ultrastructure of Endogenous Stages of Eimeria ninakohlyakimovae Yakimoff & Rastegaieff, 1930 Emend. Levine, 1961 in Experimentally Infected Goat

    Directory of Open Access Journals (Sweden)

    Vieira Luiz S

    1997-01-01

    Full Text Available The ultrastructure of endogenous stages of Eimeria ninakohlyakimovae was observed in epithelial cells of cecum and colon crypts from a goat experimentally infected with 2.0 x 105 oocysts/kg. The secondary meronts developed above the nucleus of the host cell. The nucleus first divides and merozoites then form on the surface of multinucleated meronts. Free merozoites in the parasitophorous vacuole present a conoid, double membrane, one pair of rhoptries, micronemes, micropore, anterior and posterior polar ring, a nucleus with a nucleolus and peripheral chromatin. The microgamonts are located below the nucleus of the host cell and contain several nuclei at the periphery of the parasite. The microgametes consist of a body, a nucleus, three flagella and mitochondria. The macrogamonts develop below the nucleus of the host cell and have a large nucleus with a prominent nucleolus. The macrogametes contain a nucleus, wall-forming bodies of type I and type II. The young oocysts present a wall containing two layers and a sporont

  2. Eimeria maxima microneme protein 2 delivered as DNA vaccine and recombinant protein induces immunity against experimental homogenous challenge.

    Science.gov (United States)

    Huang, Jingwei; Zhang, Zhenchao; Li, Menghui; Song, Xiaokai; Yan, Ruofeng; Xu, Lixin; Li, Xiangrui

    2015-10-01

    E. maxima is one of the seven species of Eimeria that infects chicken. Until now, only a few antigenic genes of E. maxima have been reported. In the present study, the immune protective effects against E. maxima challenge of recombinant protein and DNA vaccine encoding EmMIC2 were evaluated. Two-week-old chickens were randomly divided into five groups. The experimental group of chickens was immunized with 100 μg DNA vaccine pVAX1-MIC2 or 200 μg rEmMIC2 protein while the control group of chickens was injected with pVAX1 plasmid or sterile PBS. The results showed that the anti-EmMIC2 antibody titers of both rEmMIC2 protein and pVAX1-MIC2 groups were significantly higher as compared to PBS and pVAX1 control (Pmaxima challenge and it could be an effective antigen candidate for the development of new vaccines against E. maxima. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  3. 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. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Dietary supplementation of mannan-oligosaccharide enhances neonatal immune responses in chickens during natural exposure to Eimeria spp

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    Nava Gerardo M

    2009-03-01

    Full Text Available Abstract Background Control and eradication of intestinal infections caused by protozoa are important biomedical challenges worldwide. Prophylactic control of coccidiosis has been achieved with the use of anticoccidial drugs; however, the increase in anticoccidial resistance has raised concerns about the need for new alternatives for the control of coccidial infections. In fact, new strategies are needed to induce potent protective immune responses in neonatal individuals. Methods The effects of a dietary supplementation of mannan-oligosaccharide (yeast cell wall; YCW on the local, humoral and cell-mediated immune responses, and intestinal replication of coccidia were evaluated in a neonatal animal model during natural exposure to Eimeria spp. A total of 840 one-day-old chicks were distributed among four dietary regimens: A Control diet (no YCW plus anticoccidial vaccine; B Control diet plus coccidiostat; C YCW diet plus anticoccidial vaccination; and D YCW diet plus coccidiostat. Weight gain, feed consumption and immunological parameters were examined within the first seven weeks of life. Results Dietary supplementation of 0.05% of YCW increased local mucosal IgA secretions, humoral and cell-mediated immune responses, and reduced parasite excretion in feces. Conclusion Dietary supplementation of yeast cell wall in neonatal animals can enhance the immune response against coccidial infections. The present study reveals the potential of YCW as adjuvant for modulating mucosal immune responses.

  5. Morphological and molecular characterization of Eimeria haematodi, coccidian parasite (Apicomplexa: Eimeriidae) in a rainbow lorikeet (Trichoglossus haematodus).

    Science.gov (United States)

    Yang, Rongchang; Brice, Belinda; Ryan, Una

    2015-06-01

    Eimeria haematodi was first described in 1977 from the rainbow lorikeet (Trichoglossus haematodus) in Papua New Guinea. In the present study, we re-describe this coccidian species morphologically and molecularly from a rainbow lorikeet bird in Western Australia (WA). The oocysts were ovoid to slightly piriform and measured 28.5-37.8 by 25.8-33.0 µm (33.3 by 28.1 µm). Oocyst wall was approximately 1.5 µm thick and bilayered. Micropyle (5-7 µm) and oocyst residuum (8.0-10.0 µm) present; polar granule was absent. Sporocysts ellipsoidal, 11.8-13.6 by 8.0-9.6 µm (12.2 by 8.3 µm), with thin convex Stieda body and granular sporocyst residuum (4.0-5.0 µm). Molecular characterization of E. haematodi was conducted at 18S ribosomal RNA and the mitochondrial cytochrome oxidase gene (COI) loci. At the 18S ribosomal RNA locus, E. haematodi shared 98.1% genetic similarity to E. alabamensis from cattle in New South Wales, Australia. At COI locus, E. haematodi was closest (92.3% similarity) to E. praecox from domestic chickens (Gallus gallus domesticus) from Canada and China. Crown Copyright © 2015. Published by Elsevier Inc. All rights reserved.

  6. The life-cycle of Eimeria cernae Levine and Ivens, 1965 in the bank vole, Clethrionomys glareolus.

    Science.gov (United States)

    Lewis, D C; Ball, S J

    1982-12-01

    Eimeria cernae is recorded for the first time in England and the life-cycle is described in experimentally infected bank-voles (Clethrionomys glareolus). The pre-patent period was 6 days and the patent period was 4-6 days. Oocysts were ellipsoidal in shape and measured 20.2 x 15.9 micrometers. Sporocysts, measuring 11.5 micrometers long and 6.8 micrometers wide, possessed a small stieda body and contained a mass of granular sporocyst residuum. The endogenous stages developed in the epithelial cells of the colon and rectum. Three generations of schizonts were found. The 1st-generation schizont seen at 48 h post-infection (p.i.) contained up to 8 merozoites, the 2nd-generation schizont seen at 72 hr p.i. had a mean number of 16 (12-20) merozoites and the 3rd-generation schizont at 96 h p.i. had a mean of 18 (14-21) merozoites. Gamogonic stages were present from 96 to 120 h p.i. in the rectum only.

  7. Two COWP-like cysteine rich proteins from Eimeria nieschulzi (coccidia, apicomplexa) are expressed during sporulation and involved in the sporocyst wall formation.

    Science.gov (United States)

    Jonscher, Ernst; Erdbeer, Alexander; Günther, Marie; Kurth, Michael

    2015-07-25

    The family of cysteine rich proteins of the oocyst wall (COWPs) originally described in Cryptosporidium can also be found in Toxoplasma gondii (TgOWPs) localised to the oocyst wall as well. Genome sequence analysis of Eimeria suggests that these proteins may also exist in this genus and led us to the assumption that these proteins may also play a role in oocyst wall formation. In this study, COWP-like encoding sequences had been identified in Eimeria nieschulzi. The predicted gene sequences were subsequently utilized in reporter gene assays to observe time of expression and localisation of the reporter protein in vivo. Both investigated proteins, EnOWP2 and EnOWP6, were expressed during sporulation. The EnOWP2-promoter driven mCherry was found in the cytoplasm and the EnOWP2, respectively EnOWP6, fused to mCherry was initially observed in the extracytoplasmatic space between sporoblast and oocyst wall. This, so far unnamed compartment was designated as circumplasm. Later, the mCherry reporter co-localised with the sporocyst wall of the sporulated oocysts. This observation had been confirmed by confocal microscopy, excystation experiments and IFA. Transcript analysis revealed the intron-exon structure of these genes and confirmed the expression of EnOWP2 and EnOWP6 during sporogony. Our results allow us to assume a role, of both investigated EnOWP proteins, in the sporocyst wall formation of E. nieschulzi. Data mining and sequence comparisons to T. gondii and other Eimeria species allow us to hypothesise a conserved process within the coccidia. A role in oocyst wall formation had not been observed in E. nieschulzi.

  8. Use of pyrosequencing and denaturing gradient gel electrophoresis to examine the effects of probiotics and essential oil blends on digestive microflora in broilers under mixed Eimeria infection.

    Science.gov (United States)

    Hume, Michael E; Barbosa, Nei A; Dowd, Scot E; Sakomura, Nilva K; Nalian, Armen G; Martynova-Van Kley, Alexandra; Oviedo-Rondón, Edgar O

    2011-11-01

    A protective digestive microflora helps prevent and reduce broiler infection and colonization by enteropathogens. In the current experiment, broilers fed diets supplemented with probiotics and essential oil (EO) blends were infected with a standard mixed Eimeria spp. to determine effects of performance enhancers on ileal and cecal microbial communities (MCs). Eight treatment groups included four controls (uninfected-unmedicated [UU], unmedicated-infected, the antibiotic BMD plus the ionophore Coban as positive control, and the ionophore as negative control), and four treatments (probiotics BC-30 and Calsporin; and EO, Crina Poultry Plus, and Crina PoultryAF). Day-old broilers were raised to 14 days in floor pens on used litter and then were moved to Petersime batteries and inoculated at 15 days with mixed Eimeria spp. Ileal and cecal samples were collected at 14 days and 7 days postinfection. Digesta DNA was subjected to pyrosequencing for sequencing of individual cecal bacteria and denaturing gradient gel electrophoresis (DGGE) for determination of changes in ileal and cecal MC according to percentage similarity coefficient (%SC). Pyrosequencing is very sensitive detecting shifts in individual bacterial sequences, whereas DGGE is able to detect gross shifts in entire MC. These combined techniques offer versatility toward identifying feed additive and mild Eimeria infection modulation of broiler MC. Pyrosequencing detected 147 bacterial species sequences. Additionally, pyrosequencing revealed the presence of relatively low levels of the potential human enteropathogens Campylobacter sp. and four Shigella spp. as well as the potential poultry pathogen Clostridiun perfringens. Pre- and postinfection changes in ileal (56%SC) and cecal (78.5%SC) DGGE profiles resulted from the coccidia infection and with increased broiler age. Probiotics and EO changed MC from those seen in UU ilea and ceca. Results potentially reflect the performance enhancement above expectations in

  9. Frequency of species of the Genus Eimeria in naturally infected cattle in Southern Bahia, Northeast Brazil Frequência de espécies do gênero Eimeria em bovinos naturalmente infectados no Sudeste da Bahia, Nordeste do Brasil

    Directory of Open Access Journals (Sweden)

    Valter dos Anjos Almeida

    2011-03-01

    Full Text Available The aim of this study was to determine the presence of species of the genus Eimeria species in naturally infected bovines in Southern Bahia, Northeast Brazil. The study population comprised 117 Zebu crossbred cattle that belonged to 10 dairy herds with extensive or semi-extensive production systems. The modified Gordon and Whitlock technique was used to determine positive samples and number of oocysts per gram of feces. Statistical analyses were performed using the chi-square test with Yates correction and a 95% confidence interval. Thirty-nine cattle (33.33% were positive, and ten different species were identified in infected animals: E. bovis (24.79%; E. canadensis (8.55%; E. zuernii (6.83%; E. ellipsoidalis (5.99%; E. cylindrica (3.42%; E. auburnensis (3.42%; E. brasiliensis (2.56%; E. bukidnonensis (1.71%; E. alabamensis (0.85%, and E. subspherica (0.85%. Higher parasitism was observed in animals up to one year of age (p = 0.005, but no animal presented clinical signs of the disease. As the presence of clinical eimeriosis was not evidenced and all animals were Zebu crossbred cattle from extensive or semi-extensive production systems, further studies should be conducted to investigate the effects of these factors on disease development.O objetivo deste estudo foi determinar a presença de espécies do gênero Eimeria em bovinos naturalmente infectados, na região Sudeste da Bahia, Nordeste do Brasil. A população do estudo incluiu 117 bovinos mestiços de raças Zebuínas que pertenciam a 10 fazendas leiteiras com sistemas de produção extensivo ou semiextensivo. A técnica de Gordon e Whitlock modificada foi utilizada para determinar as amostras positivas e o número de oocistos por grama de fezes. A análise estatística foi realizada utilizando o teste do qui-quadrado com correção de Yates e intervalo de confiança de 95%. Trinta e nove animais (33,33% foram positivos, e dez diferentes espécies foram identificadas nos animais

  10. Relative disease susceptibility and clostridial toxin antibody responses in three commercial broiler lines co-infected with Clostridium perfringens and Eimeria maxima using an experimental model of necrotic enteritis

    Science.gov (United States)

    Necrotic enteritis is an enteric disease of poultry resulting from infection by Clostridium perfringens with co-infection by Eimeria spp. constituting a major risk factor for disease pathogenesis. This study compared three commercial broiler chicken lines using an experimental model of necrotic ente...

  11. 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

  12. Comparison between McMaster and Mini-FLOTAC methods for the enumeration of Eimeria maxima oocysts in poultry excreta.

    Science.gov (United States)

    Bortoluzzi, C; Paras, K L; Applegate, T J; Verocai, G G

    2018-04-30

    Monitoring Eimeria shedding has become more important due to the recent restrictions to the use of antibiotics within the poultry industry. Therefore, there is a need for the implementation of more precise and accurate quantitative diagnostic techniques. The objective of this study was to compare the precision and accuracy between the Mini-FLOTAC and the McMaster techniques for quantitative diagnosis of Eimeria maxima oocyst in poultry. Twelve pools of excreta samples of broiler chickens experimentally infected with E. maxima were analyzed for the comparison between Mini-FLOTAC and McMaster technique using, the detection limits (dl) of 23 and 25, respectively. Additionally, six excreta samples were used to compare the precision of different dl (5, 10, 23, and 46) using the Mini-FLOTAC technique. For precision comparisons, five technical replicates of each sample (five replicate slides on one excreta slurry) were read for calculating the mean oocyst per gram of excreta (OPG) count, standard deviation (SD), coefficient of variation (CV), and precision of both aforementioned comparisons. To compare accuracy between the methods (McMaster, and Mini-FLOTAC dl 5 and 23), excreta from uninfected chickens was spiked with 100, 500, 1,000, 5,000, or 10,000 OPG; additional samples remained unspiked (negative control). For each spiking level, three samples were read in triplicate, totaling nine reads per spiking level per technique. Data were transformed using log10 to obtain normality and homogeneity of variances. A significant correlation (R = 0.74; p = 0.006) was observed between the mean OPG of the McMaster dl 25 and the Mini-FLOTAC dl 23. Mean OPG, CV, SD, and precision were not statistically different between the McMaster dl 25 and Mini-FLOTAC dl 23. Despite the absence of statistical difference (p > 0.05), Mini-FLOTAC dl 5 showed a numerically lower SD and CV than Mini-FLOTAC dl 23. The Pearson correlation coefficient revealed significant and positive

  13. Re-description of a genetically typed, single oocyst line of the turkey coccidium, Eimeria dispersa Tyzzer, 1929.

    Science.gov (United States)

    El-Sherry, S; Ogedengbe, M E; Hafeez, M A; Sayf-Al-Din, M; Gad, N; Barta, J R

    2017-10-01

    The Briston strain of Eimeria dispersa Tyzzer, 1929 was isolated originally from a commercial turkey flock from Briston, Norfolk, UK. A single oocyst-derived line of E. dispersa was propagated and used to re-describe biological and morphological features of E. dispersa in the turkey. Oocysts of the Briston strain measured 26 ± 1.1 μm (24-28) by 21 ± 1 μm (19-23); these were larger than oocysts described originally by Tyzzer in 1929 (22.75 by 18.84 μm) but within dimensions (26.07 by 21.04 μm) reported by Hawkins (1952) in his description of E. dispersa isolated from turkeys. In the present study, endogenous development started mainly in duodenum and upper jejunum and then spread down toward the lower jejunum. A few parasites were detected in the ileum beginning 96 h post-infection; only few gamonts were observed in the cecal neck area at 120 h, and no parasites were detected in cecal pouches or rectum. Four asexual generations were observed before the start of gametogony, and only one large type of first generation meront was detected in duodenum and upper jejunum at 32 h. This strain has a prepatent period of 120 h. The Briston strain of E. dispersa is a mildly pathogenic coccidium. Duodenum and jejunum of infected birds were slightly dilated and paler in color than of uninfected controls. There was whitish green mucoid material in the lumen of the duodenum and jejunum. The mucosa looked slightly congested and edematous with a few scattered petechial hemorrhages.

  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

    Directory of Open Access Journals (Sweden)

    Ralph Lainson

    2002-03-01

    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.

  15. β-1,3-Glucan, Which Can Be Targeted by Drugs, Forms a Trabecular Scaffold in the Oocyst Walls of Toxoplasma and Eimeria

    Science.gov (United States)

    Bushkin, G. Guy; Motari, Edwin; Magnelli, Paula; Gubbels, Marc-Jan; Dubey, Jitender P.; Miska, Katarzyna B.; Bullitt, Esther; Costello, Catherine E.; Robbins, Phillips W.; Samuelson, John

    2012-01-01

    ABSTRACT The walls of infectious pathogens, which are essential for transmission, pathogenesis, and diagnosis, contain sugar polymers that are defining structural features, e.g., β-1,3-glucan and chitin in fungi, chitin in Entamoeba cysts, β-1,3-GalNAc in Giardia cysts, and peptidoglycans in bacteria. The goal here was to determine in which of three walled forms of Toxoplasma gondii (oocyst, sporocyst, or tissue cyst) is β-1,3-glucan, the product of glucan synthases and glucan hydrolases predicted by whole-genome sequences of the parasite. The three most important discoveries were as follows. (i) β-1,3-glucan is present in oocyst walls of Toxoplasma and Eimeria (a chicken parasite that is a model for intestinal stages of Toxoplasma) but is absent from sporocyst and tissue cyst walls. (ii) Fibrils of β-1,3-glucan are part of a trabecular scaffold in the inner layer of the oocyst wall, which also includes a glucan hydrolase that has a novel glucan-binding domain. (iii) Echinocandins, which target the glucan synthase and kill fungi, arrest development of the Eimeria oocyst wall and prevent release of the parasites into the intestinal lumen. In summary, β-1,3-glucan, which can be targeted by drugs, is an important component of oocyst walls of Toxoplasma but is not a component of sporocyst and tissue cyst walls. PMID:23015739

  16. Coccidiosis in the Chukar Partridge ( Alectoris chukar ): A Survey of Coccidiosis Outbreaks and a Test of Anticoccidial Drugs Against Eimeria kofoidi.

    Science.gov (United States)

    Gerhold, R W; Fuller, A L; McDougald, L R

    2016-12-01

    Field isolates of coccidia from 20 natural outbreaks in the chukar partridge ( Alectoris chukar ) were received from gamebird farms in 10 U.S. states. These were propagated in the laboratory and identified by microscopy and PCR. Of 20 samples, 18 were Eimeria kofoidi, two were Eimeria legionensis only, and one was a mixture of the two species. One isolate of E. kofoidi also contained an unidentified species detected only by PCR, nucleotide sequencing, and phylogenetic analysis. The efficacy of anticoccidial drugs against chukar coccidia was tested with experimental infections in battery cages. Isolates of E. kofoidi were used to infect 2-wk-old chukars. Anticoccidial products were given in the feed at levels approved for other poultry or for chukars. Tests were terminated at 6 days postinoculation with measurement of weight gains, fecal diarrhea scores, and necropsy to observe for lesion severity. Lasalocid (120 ppm) was moderately effective in one test. When tested against four field isolates, other ionophores (monensin, salinomycin, semduramicin) showed moderate effectiveness in reducing lesions and improving weight gains. Rofenaid (a potentiated sulfa mixture), robenidine (30 ppm), diclazuril (2 ppm), and decoquinate (80 ppm) were highly effective. In a test of nine products against a highly virulent field isolate, only diclazuril (2 ppm) and clopidol (125 ppm) reduced the severity of lesions and improved weight gain relative to infected controls, suggesting the extent to which previous drug usage had selected for drug resistance.

  17. Incorporation of a recombinant Eimeria maxima IMP1 antigen into nanoparticles confers protective immunity against E. Maxima challenge infection.

    Science.gov (United States)

    Jenkins, Mark C; Stevens, Laura; O'Brien, Celia; Parker, Carolyn; Miska, Katrzyna; Konjufca, Vjollca

    2018-02-14

    The purpose of this study was to determine if conjugating a recombinant Eimeria maxima protein, namely EmaxIMP1, into 20 nm polystyrene nanoparticles (NP) could improve the level of protective immunity against E. maxima challenge infection. Recombinant EmaxIMP1 was expressed in Escherichia coli as a poly-His fusion protein, purified by NiNTA chromatography, and conjugated to 20 nm polystyrene NP (NP-EmaxIMP1). NP-EMaxIMP1 or control non-recombinant (NP-NR) protein were delivered per os to newly-hatched broiler chicks with subsequent booster immunizations at 3 and 21 days of age. In battery cage studies (n = 4), chickens immunized with NP-EMaxIMP1 displayed complete protection as measured by weight gain (WG) against E. maxima challenge compared to chickens immunized with NP-NR. WG in the NP-EMaxIMP1-immunized groups was identical to WG in chickens that were not infected with E. maxima infected chickens. In floor pen studies (n = 2), chickens immunized with NP-EMaxIMP1 displayed partial protection as measured by WG against E. maxima challenge compared to chickens immunized with NP-NR. In order to understand the basis for immune stimulation, newly-hatched chicks were inoculated per os with NP-EMaxIMP1 or NP-NR protein, and the small intestine, bursa, and spleen, were examined for NP localization at 1 h and 6 h post-inoculation. Within 1 h, both NP-EMaxIMP1 and NP-NR were observed in all 3 tissues. An increase was observed in the level of NP-EmaxIMP1 and NP-NR in all tissues at 6 h post-inoculation. These data indicate that 20 nm NP-EmaxIMP1 or NP-NR reached deeper tissues within hours of oral inoculation and elicited complete to partial immunity against E. maxima challenge infection. Published by Elsevier Ltd.

  18. Three new species of coccidia (Apicomplexa: Eimeriorina) from the Marble-throated skink, Marmorosphax tricolor Bavay, 1869 (Reptilia: Scincidae), endemic to New Caledonia with a taxonomic revision of Eimeria spp. from scincid hosts

    Czech Academy of Sciences Publication Activity Database

    Modrý, David; Jirků, M.

    2006-01-01

    Roč. 99, č. 4 (2006), s. 419-428 ISSN 0932-0113 R&D Projects: GA ČR GP524/03/D104; GA ČR GA524/00/P015; GA ČR GD524/03/H133 Institutional research plan: CEZ:AV0Z60220518 Keywords : Coccdia * Reptilia * Eimeria Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 1.140, year: 2006

  19. 1480-IJBCS-Article-Ouattara Karamoko+

    African Journals Online (AJOL)

    hp

    nous avons étudié les effets de THOS sur l'indice de consommation (IC), le poids moyen des animaux, le taux ... médicaments disponibles constitue un frein ..... Kouakou SK, Touré A, Ouattara K, Djaman. AJ and N'guessan JD. 2012. In vitro anticoccidial activity of Thonningia sanguinea extract on Eimeria tenella and.

  20. Engineering of photosynthetic mannitol biosynthesis from CO2 in a cyanobacterium

    DEFF Research Database (Denmark)

    Jacobsen, Jacob Hedemand; Frigaard, Niels-Ulrik

    2014-01-01

    produce mannitol from CO2 as the sole carbon source. Two codon-optimized genes, mannitol-1-phosphate dehydrogenase (mtlD) from Escherichia coli and mannitol-1-phosphatase (mlp) from the protozoan chicken parasite Eimeria tenella, in combination encoding a biosynthetic pathway from fructose-6-phosphate...

  1. African Journal of Biotechnology - Vol 9, No 52 (2010)

    African Journals Online (AJOL)

    Oral treatment of Eimeria tenella-infected broilers using aqueous extract of wild mushroom (Ganoderma sp): Effect on haematological parameters and histopathology lesions · EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. AO Ogbe, SE Atawodi, PA Abdu, BO ...

  2. Two new species of coccidia (Apicomplexa: Eimeriidae) from leaf-tailed geckos, Uroplatus spp. (Sauria: Gekkonidae) from Madagascar, including a new host of Eimeria brygooi Upton & Barnard, 1987.

    Science.gov (United States)

    McAllister, Chris T; Scott Seville, R; Hartdegen, Ruston

    2016-10-01

    During May and June 2015, four common leaf-tailed geckos, Uroplatus fimbriatus (Schneider), five satanic leaf-tailed geckos, Uroplatus phantasticus (Boulenger), and four mossy leaf-tailed geckos, Uroplatus sikorae Boettger originally collected from Madagascar and housed at the Dallas Zoo, USA, had their faeces examined for coccidian parasites. Eight (62%) geckos were found to be passing oöcysts, including a new eimerian, a new isosporan and a previously described eimerian. Three of four (75%) U. fimbratus (type-host) and one of five (20%) U. phantasticus were infected with Eimeria schneideri n. sp.; oöcysts were subspheroidal to ellipsoidal with a bi-layered wall and measured (mean length × width, L × W) 15.1 × 13.5 µm, with a length/width (L/W) ratio of 1.1. A micropyle and oöcyst residuum were absent but one to many polar granules were present. Sporocysts were ovoidal, 6.9 × 5.3 µm, L/W = 1.3. Stieda, sub-Stieda and para-Stieda bodies were absent. A globular sporocyst residuum was present as dispersed granules. Four of five (80%) U. phantasticus harboured Isospora boulengeri n. sp.; oöcysts were subpheroidal to ellipsoidal with a bi-layered wall and measured 17.3 × 16.0 µm, L/W = 1.1. A micropyle and oöcyst residuum were absent but a polar granule was present. Sporocysts were ellipsoidal, 9.5 × 6.9 µm, L/W = 1.4. Stieda and sub-Stieda bodies were present but a para-Stieda body was absent. A globular sporocyst residuum was present with dispersed granules. In addition, one of four (25%) U. sikorae was infected with an eimerian indistinguishable from Eimeria brygooi Upton & Barnard, 1987, previously reported from Madagascar day gecko, Phelsuma grandis Gray and golddust day gecko, Phelsuma laticauda (Boettger) from Madagascar. These are the first coccidians described from Uroplatus spp.

  3. Protective efficacy of a live attenuated anti-coccidial vaccine administered to 1-day-old chickens.

    Science.gov (United States)

    Crouch, C F; Andrews, S J; Ward, R G; Francis, M J

    2003-06-01

    The efficacy of a live attenuated anti-coccidial vaccine, Paracox-5, administered to 1-day-old chicks was investigated by assessing protection against changes in weight gain following virulent challenge. Vaccinated birds were challenged independently 28 days later with each of the component species (Eimeria acervulina, Eimeria maxima, Eimeria mitis or Eimeria tenella), and protection was demonstrated against associated reduction in weight gain and lesion formation. In addition, an improvement in bird performance, in terms of feed conversion ratio, was also observed following vaccination. Furthermore, under conditions designed to more closely mimic those in the field and using hatchery spray administration, protection against a mixed virulent challenge introduced by 'seeder birds' was demonstrated evenly across a flock of broiler birds within 21 days after vaccination. These data demonstrate that Paracox-5 vaccine will protect broiler chickens against the adverse effects on performance induced by Eimeria spp.

  4. Analysis of differentially expressed genes in two immunologically distinct strains of Eimeria maxima using suppression subtractive hybridization and dot-blot hybridization

    Science.gov (United States)

    2014-01-01

    Background It is well known that different Eimeria maxima strains exhibit significant antigenic variation. However, the genetic basis of these phenotypes remains unclear. Methods Total RNA and mRNA were isolated from unsporulated oocysts of E. maxima strains SH and NT, which were found to have significant differences in immunogenicity in our previous research. Two subtractive cDNA libraries were constructed using suppression subtractive hybridization (SSH) and specific genes were further analyzed by dot-blot hybridization and qRT-PCR analysis. Results A total of 561 clones were selected from both cDNA libraries and the length of the inserted fragments was 0.25–1.0 kb. Dot-blot hybridization revealed a total of 86 differentially expressed clones (63 from strain SH and 23 from strain NT). Nucleotide sequencing analysis of these clones revealed ten specific contigs (six from strain SH and four from strain NT). Further analysis found that six contigs from strain SH and three from strain NT shared significant identities with previously reported proteins, and one contig was presumed to be novel. The specific differentially expressed genes were finally verified by RT-PCR and qRT-PCR analyses. Conclusions The data presented here suggest that specific genes identified between the two strains may be important molecules in the immunogenicity of E. maxima that may present potential new drug targets or vaccine candidates for coccidiosis. PMID:24894832

  5. CDR3 analysis of TCR Vβ repertoire of CD8⁺ T cells from chickens infected with Eimeria maxima.

    Science.gov (United States)

    Ren, Chao; Yin, Guangwen; Qin, Mei; Suo, Jingxia; Lv, Qiyao; Xie, Li; Wang, Yunzhou; Huang, Xiaoxi; Chen, Yuchen; Liu, Xianyong; Suo, Xun

    2014-08-01

    CD8(+) T cells play a major role in the immune protection of host against the reinfection of Eimeria maxima, the most immunogenic species of eimerian parasites in chickens. To explore the dominant complementarity-determining regions 3 (CDR3) of CD8(+) T cell populations induced by the infection of this parasite, sequence analysis was performed in this study for CDR3 of CD8(+) T cells from E. maxima infected chickens. After 5 days post the third or forth infection, intraepithelial lymphocytes were isolated from the jejunum of bird. CD3(+)CD8(+) T cells were sorted and subjected to total RNA isolation and cDNA preparation. PCR amplification and cloning of the loci between Vβ1 and Cβ was conducted for the subsequent sequencing of CDR3 of T cell receptor (TCR). After the forth infection, 2 birds exhibited two same frequent TCR CDR3 sequences, i.e., AKQDWGTGGYSNMI and AGRVLNIQY; while the third bird showed two different frequent TCR CDR3 sequences, AKQGARGHTPLN and AKQDIEVRGPNTPLN. No frequent CDR3 sequence was detected from uninfected birds, though AGRVLNIQY was also found in two uninfected birds. Our result preliminarily demonstrates that frequent CDR3 sequences may exist in E. maxima immunized chickens, encouraging the mining of the immunodominant CD8(+) T cells against E. maxima infection. Copyright © 2014 Elsevier Inc. All rights reserved.

  6. A new species of Eimeria Schneider, 1875 from the Serra dos Órgãos National Park, Rio de Janeiro, Brazil, with notes on its endogenous development in the montane grass mouse, Akodon montensis Thomas, 1913 (Rodentia: Sigmodontinae).

    Science.gov (United States)

    de Santana Miglionico, Marcos Tobias; Viana, Lúcio André; Barbosa, Helene Santos; Mota, Ester Maria; da Costa Neto, Sócrates Fraga; Frazão-Teixeira, Edwards; D'Andrea, Paulo Sergio

    2018-02-01

    A total of 53 specimens of the montane grass mouse, Akodon montensis Thomas, 1913 were collected in the Serra dos Órgãos National Park (SONP) in November 2014 and July 2015. The fecal material was analyzed, and a prevalence of 7.5% was recorded for a new coccidian species of the genus Eimeria Schneider, 1875, with part of its endogenous development recorded in the small intestine. The oocysts of a new coccidian species of genus Eimeria are ellipsoidal to subspherical. The wall is bi-layered, c. 1.5 μm (1.3-1.6 μm) thick, outer layer rough. Oocyst (n = 126) mean length is 25.3 μm (21.0-28.0 μm), with a width of 20.2 μm (17.0-22.0 μm) and mean length/width (L:W) ratio of 1.3 (1.2-1.4). Polar granule is present, with the oocyst residuum as a large spherical to subspherical globule. Sporocyst shape (n = 126) is ellipsoidal, with a mean length of 11.8 μm (9.3-14.4 μm), width of 7.9 μm (6.7-9.3 μm), and mean L:W ratio of 1.5 (1.4-1.7). Sporocysts with nipple-like Stieda body and sub-Stieda body are absent. A sporocyst residuum formed by several globules, usually along the sporocyst wall. This is the first record of Eimeria in the montane grass mouse from Brazil.

  7. Avaliação da eficácia da lasalocida e de alguns fatores epidemiológicos de Eimeria spp. parasitando bezerros Nelore mantidos em regime de pastejo

    Directory of Open Access Journals (Sweden)

    Leonardo Bueno Cruvinel

    Full Text Available RESUMO: A principal importância da eimeriose em bovinos, se deve ao baixo desempenho produtivo que os animais demonstram quando esta enfermidade apresenta-se sob a forma sub-clínica. Como objetivos, o presente trabalho avaliou a eficácia do uso da lasalocida sódica contra espécies de Eimeria spp. parasitando bezerros; avaliou também o desempenho ponderal dos animais submetidos aos diferentes tratamentos e analisou alguns fatores epidemiológicos que possam interferir na infecção por Eimeria nos bezerros. Foram utilizados 288 bezerros no dia 0 do estudo. Os animais pertencentes ao tratamento 01 receberam sal mineral proteinado de baixo consumo sem adição de lasalocida, enquanto que os bezerros do Tratamento 02 sal mineral proteinado de baixo consumo, com adição de lasalocida sódica, administrado via oral para bezerros dos quatro/cinco/seis meses até dez meses de idade. Colheita de fezes e pesagem dos animais foram realizadas nos dias 0 (antes do início do experimento, na desmama, 30 e 60 dias após desmama (DPD. A avaliação de alguns fatores epidemiológicos que pudessem ser relacionados com a infecção por Eimeria spp nos bezerros, como o desmame, sexo e época do ano, foram analisados neste estudo, levando-se em consideração os resultados encontrados durante todo estudo, para os 144 animais pertencentes ao grupo controle. Foram identificadas nove espécies de Eimeria nos bezerros em ordem decrescente: E. brasiliensis, E. wyomingensis, E. bovis, E. canadenses, E. zuernii, E. auburnensis, E. ellipsoidalis, E. pellita e E. cylindrica. Inesperadamente, diminuição na carga parasitária dos animais pode ser observada após o desmame. Mesmo a fazenda não adotando medidas de manejo que visam maior produtividade como a Inseminação Artificial em Tempo Fixo, que por sua vez acaba aumentando o número de nascimentos e unidade animal/hectare em uma determinada época do ano, elevado parasitismo pelo coccídio em questão foi

  8. The effect of Eimeria maxima infection on the expression of amino acid and sugar transporters aminopeptidase, as well as the di- and tri-peptide transporter PepT1, is not solely due to decreased feed intake.

    Science.gov (United States)

    Miska, Katarzyna B; Fetterer, Raymond H

    2018-05-01

    Coccidiosis caused by Eimeria in poultry is endemic to poultry operations and results in decreased feed intake, diarrhea, and decreased weight gain. The goal was to determine the effect of Eimeria maxima infection on the expression of genes that encode peptide and amino acid transporters (AATs), and also to determine whether decreased feed intake contributes to the change in gene expression by including a pair-fed group of broilers. Three groups of male Ross broilers: 1) not infected, 2) infected, and 3) not infected pair-fed groups were used. Chicks were infected with 1,000 oocysts of E. maxima at 21 d of age. Feed consumption was obtained daily, and at d 0, 3, 5, 7, 10, and 14 post-infection (PI), 6 birds were euthanized, and a portion of the jejunum was removed for qRT-PCR. Infected birds had significantly decreased feed consumption between d 6 to 9 PI. At d 7 PI infected birds had a 45% reduction in weight gain, and pair-fed birds had a 32% reduction in weight gain. The feed conversion ratio at d 7 PI of infected birds was 2.2 while that of pair-fed birds was 1.7, compared to 1.5 in uninfected birds. Growth parameters were more affected in infected birds than in pair-fed birds. By measuring expression levels of nutrient uptake and processing genes via qRT-PCR, it was determined that genes encoding proteins located at the brush border of the gut epithelium were affected by infection as well as change in feed intake. The expression of AATs B°AT, b°,+AT, EAAT3, and PepT1 in infected birds decreased sharply at the height of infection; however, in birds that were pair fed, an increase in expression of b°,+AT, and PepT1 was observed, and little change was seen in expression of B°AT and EAAT3. In summary, the changes in expression of digestive enzymes and nutrient transporters are distinct between coccidia-infected birds compared to healthy pair-fed birds.

  9. Two new species of coccidia (Apicomplexa: Eimeriidae) from leaf-tailed geckos, Uroplatus spp. (Sauria: Gekkonidae) from Madagascar, including a new host of Eimeria brygooi Upton and Barnard, 1987

    Science.gov (United States)

    McAllister, Chris T.; Seville, R. Scott; Hartdegen, Ruston

    2016-01-01

    During May and June 2015, four common leaf-tailed geckos, Uroplatus fimbriatus (Schneider), five satanic leaf-tailed geckos, Uroplatus phantasticus (Boulenger), and four mossy leaf-tailed geckos, Uroplatus sikorae Boettger originally collected from Madagascar and housed at the Dallas Zoo, USA, had their faeces examined for coccidian parasites. Eight (62%) geckos were found to be passing oöcysts, including a new eimerian, a new isosporan and a previously described eimerian. Three of four (75%) U. fimbratus (type host) and one of five (20%) U. phantasticus were infected with Eimeria schneideri n. sp.; oöcysts were subspheroidal to ellipsoidal with a bi-layered wall and measured (length × width, L × W) 15.1 × 13.5 µm, with a length/width (L/W) ratio of 1.1. A micropyle and oöcyst residuum were absent but 1–many polar granules were present. Sporocysts were ovoidal, 6.9 × 5.3 µm, L/W 1.3. Stieda, sub-Stieda and para-Stieda bodies were absent. A globular sporocyst residuum was present as dispersed granules. Four of five (80%) U. phantasticus harbored Isospora boulengeri n. sp.; oöcysts were subpheroidal to ellipsoidal with a bi-layered wall and measured 17.3 × 16.0 µm, L/W 1.1. A micropyle and oöcyst residuum were absent but a polar granule was present. Sporocysts were ellipsoidal, 9.5 × 6.9 µm, L/W 1.4. Stieda and sub-Stieda bodies were present but a para-Stieda body was absent. A globular sporocyst residuum was present with dispersed granules. In addition, one of four (25%) U. sikorae was infected with an eimerian indistinguishable from Eimeria brygooi Upton and Barnard, 1987, previously reported from Madagascar day gecko, Phelsuma grandis Gray and golddust day gecko, Phelsuma laticauda (Boettger) from Madagascar. These are the first coccidians described from Uroplatus spp. PMID:27638735

  10. Quantifying risk factors of coccidiosis in broilers using on-farm data based on a veterinary practice.

    Science.gov (United States)

    Graat, E A; van der Kooij, E; Frankena, K; Henken, A M; Smeets, J F; Hekerman, M T

    1998-01-01

    A study was done to find and quantify risk factors for coccidiosis. The study population consisted of 4774 broiler flocks kept on 177 farms. Flocks were considered a case when at least one bird in the flock showed microscopic presence of oocysts in intestinal scrapings in a grow-out cycle. Other flocks were defined as controls. This was done for three types of Eimeria: Eimeria acervulina, Eimeria tenella and Eimeria maxima. Logistic regression was used to assess variables that influence the occurrence of Eimeria species. There were 49 variables, based on animal, flock or farm level. There was an enhanced risk of coccidiosis due to environmental and management factors that increase the risk of introducing contamination or that are related to hygienic measures. These include lack of use of overalls by visitors, a farmyard which is difficult to clean, bad hygienic status, personnel who might also be working on other farms, presence of other animals on the farm, and feeding and drinking systems which are more difficult to clean. Also, the presence of other diseases on the farm and Eimeria species found in the previous flock increased the risk of coccidiosis.

  11. EIMERIOSIS IN POULTRY OF RAWALPINDI/ISLAMABAD AREA

    Directory of Open Access Journals (Sweden)

    M. Q. KHAN, H. IRSHAD, R. ANJUM, M. JAHANGIR AND U. NASIR

    2006-04-01

    Full Text Available This study was designed to determine the prevalence of eimeriosis in poultry and identify potential risk factors for its spread in Rawalpindi/Islamabad area of Pakistan. Of 359 gut samples (suspected for harbouring eimeriosis examined, 258 (71.86% were found infected. Four species of Eimeria (E. maxima, 34.10%, E. tenella, 30.62%, E. mitis, 13.95% and E. necatrix, 7.75% were recorded. The prevalence of eimeriosis was highest in the month of September (89.74%, while lowest during June (28.57%. The disease was more common at the farms where the litter was wet and not managed properly.

  12. PREVALENCE OF COCCIDIOSIS IN BROILER CHICKENS IN FAISALABAD, PAKISTAN

    Directory of Open Access Journals (Sweden)

    M. M. Ayaz, M. Akhtar, C.S. Hayat, M.A. Hafeez and A. Haq1

    2003-01-01

    Full Text Available A study was conducted during the period from October, 2000 to June, 2001 to record the prevalence of coccidiosis in broiler chickens in Faisalabad district. A total of 930 guts were collected and processed at Immunoparasitology Laboratory, Department of Veterinary Parasitology, University of Agriculture, Faisalabad. Seven species of Eimeria (E viz. E. tenella (50%, E. maxima (40%, E. mitis (2%, E. praecox (0.8%, E. acervulina (4%; E. necatrix (2% and E. brunetti (1.2% were recorded. The overall prevalence of coccidiosis was found to be 37.95%.

  13. Synthesis and SAR studies of very potent imidazopyridine antiprotozoal agents.

    Science.gov (United States)

    Biftu, Tesfaye; Feng, Dennis; Fisher, Michael; Liang, Gui-Bai; Qian, Xiaoxia; Scribner, Andrew; Dennis, Richard; Lee, Shuliang; Liberator, Paul A; Brown, Chris; Gurnett, Anne; Leavitt, Penny S; Thompson, Donald; Mathew, John; Misura, Andrew; Samaras, Samantha; Tamas, Tamas; Sina, Joseph F; McNulty, Kathleen A; McKnight, Crystal G; Schmatz, Dennis M; Wyvratt, Matthew

    2006-05-01

    Compounds 10a (IC50 110 pM) and 21 (IC50 40 pM) are the most potent inhibitors of Eimeria tenella cGMP-dependent protein kinase activity reported to date and are efficacious in the in vivo antiparasitic assay when administered to chickens at 12.5 and 6.25 ppm levels in the feed. However, both compounds are positive in the Ames microbial mutagenesis assay which precludes them from further development as antiprotozoal agents in the absence of negative lifetime rodent carcinogenicity studies.

  14. Effect of Artemisia absinthium essential oil on antioxidative systems of broiler's liver

    Directory of Open Access Journals (Sweden)

    Kostadinović Ljiljana M.

    2014-01-01

    Full Text Available The effect of Artemisia absinthium essential oil (AAEO on enzymatic activity of super-oxide-dismutase (SOD, glutathione-peroxidase (GSHPx, glutathione-reductase (GR, peroxidase (POD, xantine-oxidase (XOD and non-enzymatic (content of lipid peroxides (LPx and gluthathione (GSH antioxidative status of broilers infected with mixture of oocysts of Eimeria tenella, Eimeria mitis and Eimeria necatrix in comparison to coccidiostat salinomycin was investigated. The in vivo investigation were carried out on 120 Arbor acres broilers of both sexes. Broilers were randomly distributed into four groups. Group A was uninfected and untreated; group B was infected and was kept untreated; group C preventively received coccidiostatic salinomycin in quantity of 60 mg/kg of feed and was inoculated with coccidia species at 21st day-of-age and group D received in feed AAEO in quantity of 3 g/kg and was infected with Eimeria oocysts at 21st day-of-age. Livers were collected for the subsequent evaluation of antioxidative status. It was concluded that AAEO added in feed for broilers prevented the development of coccidia oocysts and therefore it can be used as prophylactic feed additive.

  15. Gene Discovery in the Apicomplexa as Revealed by EST Sequencing and Assembly of a Comparative Gene Database

    Science.gov (United States)

    Li, Li; Brunk, Brian P.; Kissinger, Jessica C.; Pape, Deana; Tang, Keliang; Cole, Robert H.; Martin, John; Wylie, Todd; Dante, Mike; Fogarty, Steven J.; Howe, Daniel K.; Liberator, Paul; Diaz, Carmen; Anderson, Jennifer; White, Michael; Jerome, Maria E.; Johnson, Emily A.; Radke, Jay A.; Stoeckert, Christian J.; Waterston, Robert H.; Clifton, Sandra W.; Roos, David S.; Sibley, L. David

    2003-01-01

    Large-scale EST sequencing projects for several important parasites within the phylum Apicomplexa were undertaken for the purpose of gene discovery. Included were several parasites of medical importance (Plasmodium falciparum, Toxoplasma gondii) and others of veterinary importance (Eimeria tenella, Sarcocystis neurona, and Neospora caninum). A total of 55,192 ESTs, deposited into dbEST/GenBank, were included in the analyses. The resulting sequences have been clustered into nonredundant gene assemblies and deposited into a relational database that supports a variety of sequence and text searches. This database has been used to compare the gene assemblies using BLAST similarity comparisons to the public protein databases to identify putative genes. Of these new entries, ∼15%–20% represent putative homologs with a conservative cutoff of p neurona: , , , , , , , , , , , , , –, –, –, –, –. Eimeria tenella: –, –, –, –, –, –, –, –, – , –, –, –, –, –, –, –, –, –, –, –. Neospora caninum: –, –, , – , –, –.] PMID:12618375

  16. [Epizootiological and parasitological status of the flocks at a poultry combine].

    Science.gov (United States)

    Kolev, V; Markarian, M; Genchev, G; Donev, A; Tsvetkov, S

    1976-01-01

    A comprehensive epizootiologic study has been carried out at one of the large poultry dressing combines in this country. Results have shown that the part played by infections as against the remaining diseases established in the various technologic categories is as follows: 9 to 23.8 per cent in growing parents; 16.0 to 24.6 per cent in adult parents; and 4.8 to 13.4 per cent in broilers. So far as parasitic diseases are concerned the respective figures are 3.2 to 9.9 per cent, 3.0 to 3.3. per cent, and 2.9 to 3.0 to per cent. The import of birds from France and England intended for satisfying the needs of the poultry dressing combine in Roussé has contributed to the introduction of infections which have later on assumed an epizootic course, such as infections encephalomyelitis, infectious bronchitis, and Marek' disease. The epizootic status has been aggravated with the development of a number of diseases already existing in this country--fowl pox, lymphoid leukosis, chronic respiratory disease, salmonellosis, and above all infections caused by the so-called occasionally pathogenic agents--coli organisms, staphylococci, and Pseudomonas. Of the parasitic diseases only coccidiosis has developed, being caused by a variety of species: Eimeria acervulina, Eimeria necatrix, Eimeria maxima, Eimeria muvati and Eimeria tenella. The absence of helminth infections is due to the routinely adopted technological practice of raising birds in confinement and the strict veterinary control and guard. On the basis of the results obtained and the conclusions drawn some more effective prophylactic measures are suggested correcting and supplementing the existing prophylactic programme.

  17. Chicken Coccidiosis in Central Java, Indonesia: A Recent Update.

    Science.gov (United States)

    Hamid, Penny Humaidah; Kristianingrum, Yuli Purwandari; Wardhana, April Hari; Prastowo, Sigit; da Silva, Liliana Machado Ribeiro

    2018-01-01

    Avian coccidiosis is a huge problem worldwide. Heavily infected animals that show severe clinical signs and coccidiostat resistance are causing important economic losses. The present study aimed to update the recent cases of coccidiosis in Central Java, Indonesia, and to show the importance of the disease in the region. A total of 699 samples were obtained from different chicken breed. Different Eimeria species were detected in 175 individuals (25.04%). Three different groups of chicken breed were considered: local chicken (autochthonous chickens of Sentul and Jawa), commercial broiler, and layer. Broiler chickens showed the highest prevalence of infection (34%), followed by layer (26.26%) and local chickens (10.45%). Mild to severe clinical signs of avian coccidiosis were observed in 42% of the infected animals, while 58% of the infected animals showed no clinical signs other than low feed conversion rates. Seven different Eimeria species were identified: E. tenella was the most prevalent (43.3%), followed by E. maxima (26.3%), E. necatrix (15.7%), E. acervulina (8%), E. praecox (3.1%), E. mitis (2.2%), and E. brunetti (1.3%). Coinfections with several Eimeria species were diagnosed. With this study we found massive usage of coccidiostat in the region even though its usage cannot guarantee coccidiosis-free chicken production.

  18. Effect of micelle interface on the binding of anticoccidial PW2 peptide

    International Nuclear Information System (INIS)

    Tinoco, Luzineide W.; Gomes-Neto, Francisco; Valente, Ana Paula; Almeida, Fabio C. L.

    2007-01-01

    PW2 is an anticoccidial peptide active against Eimeria acervulina and Eimeria tenella. We determined the structure of PW2 in dodecylphosphocholine micelles. The structure showed two distinct regions: an amphipathic N-terminal 3 10 helix and an aromatic region containing WWR interface-binding motif. The aromatic region acted as a scaffold of the protein in the interface and shared the same structure in both DPC and SDS micelles. N-terminal helix interacted with DPC but not with SDS interface. Chemical shift change was slow when SDS was added to PW2 in DPC and fast when DPC was added to PW2 in SDS, indicating that interaction with DPC micelles was kinetically more stable than with SDS micelles. Also, DPC interface was able to accommodate PW2, but it maintained the conformational arrangement in the aromatic region observed for SDS micelles. This behavior, which is different from that observed for other antimicrobial peptides with WWR motif, may be associated with the absence of PW2 antibacterial activity and its selectivity for Eimeria parasites

  19. Effect of micelle interface on the binding of anticoccidial PW2 peptide

    Energy Technology Data Exchange (ETDEWEB)

    Tinoco, Luzineide W. [Universidade Federal do Rio de Janeiro, Nucleo de Pesquisas de Produtos Naturais (Brazil); Gomes-Neto, Francisco; Valente, Ana Paula; Almeida, Fabio C. L. [Universidade Federal do Rio de Janeiro, Centro Nacional de Ressonancia Magnetica Nuclear Jiri Jonas, Instituto de Bioquimica Medica, Programa de Biologia Estrutural (Brazil)], E-mail: falmeida@cnrmn.bioqmed.ufrj.br

    2007-12-15

    PW2 is an anticoccidial peptide active against Eimeria acervulina and Eimeria tenella. We determined the structure of PW2 in dodecylphosphocholine micelles. The structure showed two distinct regions: an amphipathic N-terminal 3{sub 10} helix and an aromatic region containing WWR interface-binding motif. The aromatic region acted as a scaffold of the protein in the interface and shared the same structure in both DPC and SDS micelles. N-terminal helix interacted with DPC but not with SDS interface. Chemical shift change was slow when SDS was added to PW2 in DPC and fast when DPC was added to PW2 in SDS, indicating that interaction with DPC micelles was kinetically more stable than with SDS micelles. Also, DPC interface was able to accommodate PW2, but it maintained the conformational arrangement in the aromatic region observed for SDS micelles. This behavior, which is different from that observed for other antimicrobial peptides with WWR motif, may be associated with the absence of PW2 antibacterial activity and its selectivity for Eimeria parasites.

  20. Genetic engineering of cyanobacteria

    DEFF Research Database (Denmark)

    Jacobsen, Jacob Hedemand

    , was assembled in our novel USER-cloning based expression vector. The construct was inserted in the cyanobacterial chromosome and expressed resulting in accumulation of mannitol in the cells and in the culture medium. Insertion of the biosynthetic pathway in the glycogen synthase double mutant increased....... For that purpose we identified an irreversible two-step biosynthetic pathway from fructose-6-phosphate to mannitol. The pathway, comprised by mannitol-1-phosphate dehydrogenase (mtlD) from Escherichia coli and codon-optimized mannitol-1-phosphatase (mlp) from the protozoan chicken parasite Eimeria tenella...... into the cyanobacterial chromosome as well as the genotype of the parental strain. This was exemplified by the fact that production of mannitol from the parental strain, deficient in glycogen biosynthesis, was 10 times higher than from wild type parental background. The combined intra and extracellular mannitol yield...

  1. Fudecalone, a new anticoccidial agent produced by Penicillium sp. FO-2030.

    Science.gov (United States)

    Tabata, N; Tomoda, H; Masuma, R; Iwai, Y; Omura, S

    1995-01-01

    Penicillium sp. FO-2030, a soil isolate, was found to produce a new anticoccidial compound. The active compound, designated fudecalone, was isolated from the fermentation broth of the producing strain by solvent extraction, silica gel column chromatography and preparative HPLC. The structure of fudecalone was elucidated to be 3,3a,6,6a,7,8,9,10-octahydro-1-hydroxy-4,7,7-trimethyl-1H-naphtho[1,8a- c]furan-6-one mainly by spectroscopic studies including various NMR measurements. The anticoccidial activity using cell systems indicated that schizont formation of monensin-resistant Eimeria tenella was completely inhibited by fudecalone at concentrations more than 16 microM.

  2. Arohynapenes A and B, new anticoccidial agents produced by Penicillium sp. Taxonomy, fermentation, and structure elucidation.

    Science.gov (United States)

    Masuma, R; Tabata, N; Tomoda, H; Haneda, K; Iwai, Y; Omura, S

    1994-01-01

    Penicillium sp. FO-2295, a water isolate, was found to produce a series of new anticoccidial compounds. Two active compounds, designated arohynapenes A and B, were isolated from the fermentation broth of the producing strain by solvent extration and preparative HPLC. Arohynapene A was deduced to be (2E,4E)-5-(5-hydroxy-2,6,8-trimethyl-5,6,7,8-tetrahydronaphtale ne)-2,4- pentadienoic acid, and arohynapene B was (2E,4E)-5-(2-hydroxymethyl-6,8-dimethyl-5,6,7,8-tetrahydronapht alene)-2,4- pentadienoic acid. Arohynapenes inhibited the growth of Eimeria tenella in an in vitro assay using BHK-21 cells as a host. No schizont in the cells was observed at concentrations ranging above 35.0 microM and 7.0 microM for arohynapenes A and B, respectively.

  3. Carotenoid-enriched transgenic corn delivers bioavailable carotenoids to poultry and protects them against coccidiosis.

    Science.gov (United States)

    Nogareda, Carmina; Moreno, Jose A; Angulo, Eduardo; Sandmann, Gerhard; Portero, Manuel; Capell, Teresa; Zhu, Changfu; Christou, Paul

    2016-01-01

    Carotenoids are health-promoting organic molecules that act as antioxidants and essential nutrients. We show that chickens raised on a diet enriched with an engineered corn variety containing very high levels of four key carotenoids (β-carotene, lycopene, zeaxanthin and lutein) are healthy and accumulate more bioavailable carotenoids in peripheral tissues, muscle, skin and fat, and more retinol in the liver, than birds fed on standard corn diets (including commercial corn supplemented with colour additives). Birds were challenged with the protozoan parasite Eimeria tenella and those on the high-carotenoid diet grew normally, suffered only mild disease symptoms (diarrhoea, footpad dermatitis and digital ulcers) and had lower faecal oocyst counts than birds on the control diet. Our results demonstrate that carotenoid-rich corn maintains poultry health and increases the nutritional value of poultry products without the use of feed additives. © 2015 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

  4. Evaluation of Montanide™ ISA 71 VG adjuvant during profilin vaccination against experimental coccidiosis.

    Directory of Open Access Journals (Sweden)

    Seung I Jang

    Full Text Available Chickens were immunized subcutaneously with an Eimeria recombinant profilin protein plus Montanide™ ISA 70 VG (ISA 70 or Montanide™ ISA 71 VG (ISA 71 water-in-oil adjuvants, or with profilin alone, and comparative RNA microarray hybridizations were performed to ascertain global transcriptome changes induced by profilin/ISA 70 vs. profilin alone and by profilin/ISA 71 vs. profilin alone. While immunization with profilin/ISA 70 vs. profilin alone altered the levels of more total transcripts compared with profilin/ISA 71 vs. profilin alone (509 vs. 296, the latter was associated with a greater number of unique biological functions, and a larger number of genes within these functions, compared with the former. Further, canonical pathway analysis identified 10 pathways that were associated with genes encoding the altered transcripts in animals immunized with profilin/ISA 71 vs. profilin alone, compared with only 2 pathways in profilin/ISA 70 vs. profilin alone. Therefore, ISA 71 was selected as a candidate adjuvant in conjunction with profilin vaccination for in vivo disease protection studies. Vaccination with profilin/ISA 71 was associated with greater body weight gain following E. acervulina infection, and decreased parasite fecal shedding after E. maxima infection, compared with profilin alone. Anti-profilin antibody levels were higher in sera of E. maxima- and E. tenella-infected chickens vaccinated with profilin/ISA 71 compared with profilin alone. Finally, the levels of transcripts encoding interferon-γ, interleukin (IL-2, IL-10, and IL-17A were increased in intestinal lymphocytes from E. acervulina-, E. maxima-, and/or E. tenella-infected chickens vaccinated with profilin/ISA 71 compared with profilin alone. None of these effects were seen in chickens injected with ISA 71 alone indicating that the adjuvant was not conferring non-specific immune stimulation. These results suggest that profilin plus ISA 71 augments protective immunity

  5. Evaluation of Montanide™ ISA 71 VG Adjuvant during Profilin Vaccination against Experimental Coccidiosis

    Science.gov (United States)

    Lillehoj, Hyun S.; Lee, Sung Hyen; Lee, Kyung Woo; Bertrand, François; Dupuis, Laurent; Deville, Sébastien; Ben Arous, Juliette; Lillehoj, Erik P.

    2013-01-01

    Chickens were immunized subcutaneously with an Eimeria recombinant profilin protein plus Montanide™ ISA 70 VG (ISA 70) or Montanide™ ISA 71 VG (ISA 71) water-in-oil adjuvants, or with profilin alone, and comparative RNA microarray hybridizations were performed to ascertain global transcriptome changes induced by profilin/ISA 70 vs. profilin alone and by profilin/ISA 71 vs. profilin alone. While immunization with profilin/ISA 70 vs. profilin alone altered the levels of more total transcripts compared with profilin/ISA 71 vs. profilin alone (509 vs. 296), the latter was associated with a greater number of unique biological functions, and a larger number of genes within these functions, compared with the former. Further, canonical pathway analysis identified 10 pathways that were associated with genes encoding the altered transcripts in animals immunized with profilin/ISA 71 vs. profilin alone, compared with only 2 pathways in profilin/ISA 70 vs. profilin alone. Therefore, ISA 71 was selected as a candidate adjuvant in conjunction with profilin vaccination for in vivo disease protection studies. Vaccination with profilin/ISA 71 was associated with greater body weight gain following E. acervulina infection, and decreased parasite fecal shedding after E. maxima infection, compared with profilin alone. Anti-profilin antibody levels were higher in sera of E. maxima- and E. tenella-infected chickens vaccinated with profilin/ISA 71 compared with profilin alone. Finally, the levels of transcripts encoding interferon-γ, interleukin (IL)-2, IL-10, and IL-17A were increased in intestinal lymphocytes from E. acervulina-, E. maxima-, and/or E. tenella-infected chickens vaccinated with profilin/ISA 71 compared with profilin alone. None of these effects were seen in chickens injected with ISA 71 alone indicating that the adjuvant was not conferring non-specific immune stimulation. These results suggest that profilin plus ISA 71 augments protective immunity against selective

  6. Multidrug ATP-binding cassette transporters are essential for hepatic development of Plasmodium sporozoites

    NARCIS (Netherlands)

    Rijpma, S.R.; Velden, M. van der; Gonzalez-Pons, M.; Annoura, T.; Schaijk, B.C.L. van; Gemert, G.J.A. van; Heuvel, J.M.W. van den; Ramesar, J.; Chevalley-Maurel, S.; Ploemen, I.H.; Khan, S.M.; Franetich, J.F.; Mazier, D.; Wilt, J.H.W. de; Serrano, A.E.; Russel, F.G.; Janse, C.J.; Sauerwein, R.W.; Koenderink, J.B.; Franke-Fayard, B.M.

    2016-01-01

    Multidrug resistance-associated proteins (MRPs) belong to the C-family of ATP-binding cassette (ABC) transport proteins and are known to transport a variety of physiologically important compounds and to be involved in the extrusion of pharmaceuticals. Rodent malaria parasites encode a single ABC

  7. Multidrug ATP-binding cassette transporters are essential for hepatic development of Plasmodium sporozoites.

    Science.gov (United States)

    Rijpma, Sanna R; van der Velden, Maarten; González-Pons, Maria; Annoura, Takeshi; van Schaijk, Ben C L; van Gemert, Geert-Jan; van den Heuvel, Jeroen J M W; Ramesar, Jai; Chevalley-Maurel, Severine; Ploemen, Ivo H; Khan, Shahid M; Franetich, Jean-Francois; Mazier, Dominique; de Wilt, Johannes H W; Serrano, Adelfa E; Russel, Frans G M; Janse, Chris J; Sauerwein, Robert W; Koenderink, Jan B; Franke-Fayard, Blandine M

    2016-03-01

    Multidrug resistance-associated proteins (MRPs) belong to the C-family of ATP-binding cassette (ABC) transport proteins and are known to transport a variety of physiologically important compounds and to be involved in the extrusion of pharmaceuticals. Rodent malaria parasites encode a single ABC transporter subfamily C protein, whereas human parasites encode two: MRP1 and MRP2. Although associated with drug resistance, their biological function and substrates remain unknown. To elucidate the role of MRP throughout the parasite life cycle, Plasmodium berghei and Plasmodium falciparum mutants lacking MRP expression were generated. P. berghei mutants lacking expression of the single MRP as well as P. falciparum mutants lacking MRP1, MRP2 or both proteins have similar blood stage growth kinetics and drug-sensitivity profiles as wild type parasites. We show that MRP1-deficient parasites readily invade primary human hepatocytes and develop into mature liver stages. In contrast, both P. falciparum MRP2-deficient parasites and P. berghei mutants lacking MRP protein expression abort in mid to late liver stage development, failing to produce mature liver stages. The combined P. berghei and P. falciparum data are the first demonstration of a critical role of an ABC transporter during Plasmodium liver stage development. © 2015 John Wiley & Sons Ltd.

  8. Controlled Human Malaria Infection of Tanzanians by Intradermal Injection of Aseptic, Purified, Cryopreserved Plasmodium falciparum Sporozoites

    NARCIS (Netherlands)

    Shekalaghe, S.; Rutaihwa, M.; Billingsley, P.F.; Chemba, M.; Daubenberger, C.A.; James, E.R.; Mpina, M.; Juma, O. Ali; Schindler, T.; Huber, E.; Gunasekera, A.; Manoj, A.; Simon, B.; Saverino, E.; Church, L.W.; Hermsen, C.C.; Sauerwein, R.W.; Plowe, C.; Venkatesan, M.; Sasi, P.; Lweno, O.; Mutani, P.; Hamad, A.; Mohammed, A.; Urassa, A.; Mzee, T.; Padilla, D.; Ruben, A.; Sim, B.K.; Tanner, M.; Abdulla, S.; Hoffman, S.L.

    2014-01-01

    Controlled human malaria infection (CHMI) by mosquito bite has been used to assess anti-malaria interventions in > 1,500 volunteers since development of methods for infecting mosquitoes by feeding on Plasmodium falciparum (Pf) gametocyte cultures. Such CHMIs have never been used in Africa. Aseptic,

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

    Science.gov (United States)

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

    1999-04-01

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

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

    OpenAIRE

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

    1999-01-01

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

  11. Immune Responses and Protection of Aotus Monkeys Immunized with Irradiated Plasmodium vivax Sporozoites

    Science.gov (United States)

    2011-01-01

    The experimental protocol was approved by the Animal Ethical Committee of the Universidad del Valle (Cali). Parasite and irradiation. Plasmodium...three repeats ofpll (GDRADGPA) and (ANGAGNQPG) sequences, derived from VK210 and VK247 CS variants, respectively. A non-malaria related peptide Ptt30...Universidad del Valle and Centro Internacional de Vacunas, Cali, Colombia, E-mails: alejovi@hotmail.com, anbonelo@yahoo.com, alejcaste@yahoo.com

  12. Fine structure of endogenous stages of Eimeria turcicus developing ...

    African Journals Online (AJOL)

    1988-05-06

    . G: Golgi. GC: Granular cisterna. g: Microgamete. gb: Gall bladder epithelium. H: Host cell. hn: Host nucleus. L: Lipid vacuoles. MI-5: Pellicular envelopes of the oocyst. MA: Macrogamont. MI: Microgamont. Mr: Meront. Mz:.

  13. Prevalence of Eimeria species in local breed chickens in Gombe ...

    African Journals Online (AJOL)

    2Department of Biological Sciences, Gombe State University, Gombe, PMB 127, ... young birds 34 (56.7%) and birds from the free range management system 44 ..... study as her undergraduate student project, .... Animale, Puteaux France, 12.

  14. 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.

  15. Prevalence of Coccidiosis in Free-Range Chicken in Sidi Thabet, Tunisia

    Directory of Open Access Journals (Sweden)

    Khaled Kaboudi

    2016-01-01

    Full Text Available Background. Enteric diseases are an important concern to the poultry industry and coccidiosis is imposing a significant economic burden worldwide. Objectives. The main goal of the present study was to investigate the prevalence of coccidiosis in free-range chicken in Sidi Thabet, northeast Tunisia. Methods. Six hundred and thirty free-range chickens along with fecal samples were collected from 15 flocks in this region and two hundred chickens were found positive for oocysts of Eimeria spp. Intestines were dissected and examined for macroscopic lesions. The mucosa of small intestine and the caeca were examined for the presence and identification of parasitic forms using parasitology methods. The mean lesion scores were usually low (2+ were attributed mainly to the caeca. Results. The overall rate of coccidiosis was 31.8%: E. tenella (61.5%, E. maxima (12%, and E. acervulina (1.5%. Mixed Eimeria species infection was observed with overall prevalence 26.5%. There was a statistically significant difference (P<0.05 among infection rates, age groups, season, diarrhea, and type of chicken. Conclusion. This is the first report of coccidiosis rate in free-range chicken in this region. Further additional studies are needed to develop better preventive measures against coccidiosis in the country.

  16. Prevalence of Coccidiosis in Free-Range Chicken in Sidi Thabet, Tunisia.

    Science.gov (United States)

    Kaboudi, Khaled; Umar, Sajid; Munir, Muhammad Tanveer

    2016-01-01

    Background. Enteric diseases are an important concern to the poultry industry and coccidiosis is imposing a significant economic burden worldwide. Objectives. The main goal of the present study was to investigate the prevalence of coccidiosis in free-range chicken in Sidi Thabet, northeast Tunisia. Methods. Six hundred and thirty free-range chickens along with fecal samples were collected from 15 flocks in this region and two hundred chickens were found positive for oocysts of Eimeria spp. Intestines were dissected and examined for macroscopic lesions. The mucosa of small intestine and the caeca were examined for the presence and identification of parasitic forms using parasitology methods. The mean lesion scores were usually low (2+) were attributed mainly to the caeca. Results. The overall rate of coccidiosis was 31.8%: E. tenella (61.5%), E. maxima (12%), and E. acervulina (1.5%). Mixed Eimeria species infection was observed with overall prevalence 26.5%. There was a statistically significant difference (P free-range chicken in this region. Further additional studies are needed to develop better preventive measures against coccidiosis in the country.

  17. Antibody expressing pea seeds as fodder for prevention of gastrointestinal parasitic infections in chickens

    Directory of Open Access Journals (Sweden)

    Macek Jeanette

    2009-09-01

    Full Text Available Abstract Background Coccidiosis caused by protozoans of genus Eimeria is a chicken parasitic disease of great economical importance. Conventional disease control strategies depend on vaccination and prophylactic use of anticoccidial drugs. Alternative solution to prevent and treat coccidiosis could be provided by passive immunization using orally delivered neutralizing antibodies. We investigated the possibility to mitigate the parasitic infection by feeding poultry with antibody expressing transgenic crop seeds. Results Using the phage display antibody library, we generated a panel of anti-Eimeria scFv antibody fragments with high sporozoite-neutralizing activity. These antibodies were expressed either transiently in agrobacteria-infiltrated tobacco leaves or stably in seeds of transgenic pea plants. Comparison of the scFv antibodies purified either from tobacco leaves or from the pea seeds demonstrated no difference in their antigen-binding activity and molecular form compositions. Force-feeding experiments demonstrated that oral delivery of flour prepared from the transgenic pea seeds had higher parasite neutralizing activity in vivo than the purified antibody fragments isolated from tobacco. The pea seed content was found to protect antibodies against degradation by gastrointestinal proteases (>100-fold gain in stability. Ad libitum feeding of chickens demonstrated that the transgenic seeds were well consumed and not shunned. Furthermore, feeding poultry with shred prepared from the antibody expressing pea seeds led to significant mitigation of infection caused both by high and low challenge doses of Eimeria oocysts. Conclusion The results suggest that our strategy offers a general approach to control parasitic infections in production animals using cost-effective antibody expression in crop seeds affordable for the animal health market.

  18. Development of a novel subunit vaccine against East Coast fever based on the Theileria parva sporozoite surface protein p67

    NARCIS (Netherlands)

    Kaba, S.A.

    2003-01-01

    Theileriaparva is an intracellular protozoan parasite and the causative agent of a lethal cattle disease, called East Coast fever (ECF). This disease poses a major constraint on improvement of cattle production in Eastern, Central

  19. Adenovirus Particles that Display the Plasmodium falciparum Circumsporozoite Protein NANP Repeat Induce Sporozoite-Neutralizing Antibodies in Mice

    OpenAIRE

    Palma, Christopher; Overstreet, Michael G.; Guedon, Jean-Marc; Hoiczyk, Egbert; Ward, Cameron; Karen, Kasey A.; Zavala, Fidel; Ketner, Gary

    2011-01-01

    Adenovirus particles can be engineered to display exogenous peptides on their surfaces by modification of viral capsid proteins, and particles that display pathogen-derived peptides can induce protective immunity. We constructed viable recombinant adenoviruses that display B-cell epitopes from the Plasmodium falciparum circumsporozoite protein (PfCSP) in the major adenovirus capsid protein, hexon. Recombinants induced high-titer antibodies against CSP when injected intraperitoneally into mice...

  20. Structure-activity-based design of a synthetic malaria peptide eliciting sporozoite inhibitory antibodies in a virosomal formulation.

    NARCIS (Netherlands)

    Okitsu, S.L.; Kienzl, U.; Moehle, K.; Silvie, O.; Peduzzi, E.; Mueller, M.S.; Sauerwein, R.W.; Matile, H.; Zurbriggen, R.; Mazier, D.; Robinson, J.A.; Pluschke, G.

    2007-01-01

    The circumsporozoite protein (CSP) of Plasmodium falciparum is a leading candidate antigen for inclusion in a malaria subunit vaccine. We describe here the design of a conformationally constrained synthetic peptide, designated UK-39, which has structural and antigenic similarity to the NPNA-repeat

  1. Early Transcriptional Responses of HepG2-A 16 Liver Cells to Infection by Plasmodium falciparum Sporozoites

    Science.gov (United States)

    2011-07-29

    related protein complex 3, delta l subunit Solute carrier family 25, member 36 RAP! interacting factor homolog ( yeast ) Topoisomerase (DNA) I...virus (HCV) showed strong staining for glypican-3 (51), and yeast two-hybrid assays revealed that glypican-3 interacts with CD8l, a member of the...jenwithisuk, R., Coleman, R. E., Udomsangpetch, R., Cui, L., and Brewer , T. G. (2006) Am. f. Trap. Med. Hyg. 74, 708-715 18. Albuquerque, S. S., Carrel, C

  2. Widespread occurrence of organelle genome-encoded 5S rRNAs including permuted molecules.

    Science.gov (United States)

    Valach, Matus; Burger, Gertraud; Gray, Michael W; Lang, B Franz

    2014-12-16

    5S Ribosomal RNA (5S rRNA) is a universal component of ribosomes, and the corresponding gene is easily identified in archaeal, bacterial and nuclear genome sequences. However, organelle gene homologs (rrn5) appear to be absent from most mitochondrial and several chloroplast genomes. Here, we re-examine the distribution of organelle rrn5 by building mitochondrion- and plastid-specific covariance models (CMs) with which we screened organelle genome sequences. We not only recover all organelle rrn5 genes annotated in GenBank records, but also identify more than 50 previously unrecognized homologs in mitochondrial genomes of various stramenopiles, red algae, cryptomonads, malawimonads and apusozoans, and surprisingly, in the apicoplast (highly derived plastid) genomes of the coccidian pathogens Toxoplasma gondii and Eimeria tenella. Comparative modeling of RNA secondary structure reveals that mitochondrial 5S rRNAs from brown algae adopt a permuted triskelion shape that has not been seen elsewhere. Expression of the newly predicted rrn5 genes is confirmed experimentally in 10 instances, based on our own and published RNA-Seq data. This study establishes that particularly mitochondrial 5S rRNA has a much broader taxonomic distribution and a much larger structural variability than previously thought. The newly developed CMs will be made available via the Rfam database and the MFannot organelle genome annotator. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  3. The destruction of parasitic resistant stages in sewage sludge by irradiation with low accelerating voltage electrons

    International Nuclear Information System (INIS)

    Enigk, K.; Holl, P.; Dey-Hazra, A.; Polymer-Physik G.m.b.H. und Co. K.G., Tuebingen

    1975-01-01

    The destroying effect of ionizing radiation on parasitic resistant stages in sludge has been tested. Suitable for that process is an electron beam accelerator which will be provided with energy from the electric power supply network which can be switched on and off according to the requirements. Such modern utilities have an enormous beam capacity and a high operating safety. The process is working according to the continuous flow principle and at room temperature. In a series of 13 experiments the effect of different doses has been tested. A dose of 480 kRad (accelerating voltage: 400 kV, beam current: 10 mA , irradiation time: 24 sec.) can easily obtained in practical work and is economically acceptable. By these means approximately 97% of the following parasitic stages have been destroyed: undeveloped eggs of Ascaris suum, Trichuris suis, Fasciola hepatica and gastrointestinal strongylids of pigs, embryonated eggs of Capillaria obsignata and probably of Taenia spec. A few third-stage larvae of Oesophagostomum (Strongylidae) of pigs survived even 108 sec of irradiation; however, they did not develop to maturity in the definitive host. Approximately 25% of the sporulated oocysts of Eimeria tenella were still infective after 108 sec of irradiation. (orig.) [de

  4. 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.

  5. Enhancement of the anticoccidial activity of polyether antibiotics in chickens by tiamulin.

    Science.gov (United States)

    Meingassner, J G; Schmook, F P; Czok, R; Mieth, H

    1979-03-01

    The anticoccidial activities of monensin and lasalocid have been studied separately and in combination with tiamulin, a new pleuromutilin derivative. Combinations of constant tiamulin concentration (.0125%) in drinking water with various levels of polyether anticoccidials (6.3 to 125 ppm) in feed and conversely of constant levels of anticoccidials with various concentrations of tiamulin were used. The prophylactic efficacy of these combined treatments in battery raised broiler chickens infected with Eimeria tenella was evaluated. Assessment of the parameters mortality, weight gain, dropping scores, lesion scores, and oocyst output showed that simultaneous application of tiamulin significantly improved the anticoccidial activity of the polyethers. As tiamulin alone is without anticoccidial activity, this phenomenon was considered to result from an interaction between tiamulin and the polyethers leading to a slower metabolic degradation of the latter. Thus tissue levels adequate for maximum anticoccidial activity would be attained with lower polyether dose levels. Experiments using isolated perfused rat liver showed that elimination of monensin was reduced by 60% in the presence of tiamulin.

  6. Analysis of the function of IL-10 in chickens using specific neutralising antibodies and a sensitive capture ELISA.

    Science.gov (United States)

    Wu, Zhiguang; Hu, Tuanjun; Rothwell, Lisa; Vervelde, Lonneke; Kaiser, Pete; Boulton, Kay; Nolan, Matthew J; Tomley, Fiona M; Blake, Damer P; Hume, David A

    2016-10-01

    In mammals, the inducible cytokine interleukin 10 is a feedback negative regulator of inflammation. To determine the extent to which this function is conserved in birds, recombinant chicken IL-10 was expressed as a secreted human Ig Fc fusion protein (chIL-10-Fc) and used to immunise mice. Five monoclonal antibodies (mAb) which specifically recognise chicken IL-10 were generated and characterised. Two capture ELISA assays were developed which detected native chIL-10 secreted from chicken bone marrow-derived macrophages (chBMMs) stimulated with lipopolysaccharide (LPS). Three of the mAbs detected intracellular IL-10. This was detected in only a subset of the same LPS-stimulated chBMMs. The ELISA assay also detected massive increases in circulating IL-10 in chickens challenged with the coccidial parasite, Eimeria tenella. The same mAbs neutralised the bioactivity of recombinant chIL-10. The role of IL-10 in feedback control was tested in vitro. The neutralising antibodies prevented IL-10-induced inhibition of IFN-γ synthesis by mitogen-activated lymphocytes and increased nitric oxide production in LPS-stimulated chBMMs. The results confirm that IL-10 is an inducible feedback regulator of immune response in chickens, and could be the target for improved vaccine efficacy or breeding strategies. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  7. Coccidiosis and performance in broilers with anticoccidial medicated feed starting at different ages

    Directory of Open Access Journals (Sweden)

    Costa C.A.F.

    2000-01-01

    Full Text Available The effects of age, litter reutilization, and anticoccidial programs were studied in broilers. The oocyst counts increased with litter reutilization up to third use, and T1 (no anticoccidial drug over the entire 42-day-period and T4 (anticoccidial medication starting from the first day showed the highest counts. The scores for Eimeria acervulina increased up to third use of litter and declined by the fourth use. The scores for E. maxima increased from first to second litter use, and from this to third and fourth use. The scores for E. tenella increased from first and second litter use to third litter use, and declined on fourth use. With high coccidial contamination, as in third use litter, the anticoccidial medication starting from the 1st, 7th (T3 and 14th (T2 day of age presented similar results of lesion scores and average weight gain. When coccidial contamination was low or moderate, as in first, second and fourth use litter, the anticoccidial medication starting at 14 days of age tended to result in a better broiler performance. These data suggest that, with low coccidial challenge, as in broilers raised in first use litter and in farms with single bird age, the delayed anticoccidial medication may provide advantages over the full medication starting from the first day of age.

  8. The establishment of Saccharomyces boulardii surface display system using a single expression vector.

    Science.gov (United States)

    Wang, Tiantian; Sun, Hui; Zhang, Jie; Liu, Qing; Wang, Longjiang; Chen, Peipei; Wang, Fangkun; Li, Hongmei; Xiao, Yihong; Zhao, Xiaomin

    2014-03-01

    In the present study, an a-agglutinin-based Saccharomyces boulardii surface display system was successfully established using a single expression vector. Based on the two protein co-expression vector pSP-G1 built by Partow et al., a S. boulardii surface display vector-pSDSb containing all the display elements was constructed. The display results of heterologous proteins were confirmed by successfully displaying enhanced green fluorescent protein (EGFP) and chicken Eimeria tenella Microneme-2 proteins (EtMic2) on the S. boulardii cell surface. The DNA sequence of AGA1 gene from S. boulardii (SbAGA1) was determined and used as the cell wall anchor partner. This is the first time heterologous proteins have been displayed on the cell surface of S. boulardii. Because S. boulardii is probiotic and eukaryotic, its surface display system would be very valuable, particularly in the development of a live vaccine against various pathogenic organisms especially eukaryotic pathogens such as protistan parasites. Copyright © 2013 Elsevier Inc. All rights reserved.

  9. Antibacterial, antiviral, and antifungal properties of wines and winery byproducts in relation to their flavonoid content.

    Science.gov (United States)

    Friedman, Mendel

    2014-07-02

    Grapes produce organic compounds that may be involved in the defense of the plants against invading phytopathogens. These metabolites include numerous phenolic compounds that are also active against human pathogens. Grapes are used to produce a variety of wines, grape juices, and raisins. Grape pomace, seeds, and skins, the remains of the grapes that are a byproduct of winemaking, also contain numerous bioactive compounds that differ from those found in grapes and wines. This overview surveys and interprets our present knowledge of the activities of wines and winery byproducts and some of their bioactive components against foodborne (Bacillus cereus, Campylobacter jejuni, Escherichia coli, Listeria monocytogenes, Salmonella enterica, Staphylococcus aureus, Yersinia enterocolitica, Vibrio cholerae, Vibrio vulnificus), medical (Helicobacter pylori, Klebsiella pneumoniae), and oral pathogenic bacteria, viruses (adeno, cytomegalo, hepatitis, noro, rota), fungi (Candida albicans, Botrytis cinerea), parasites (Eimeria tenella, Trichomonas vaginalis), and microbial toxins (ochratoxin A, Shiga toxin) in culture, in vivo, and in/on food (beef, chicken, frankfurters, hot dogs, lettuce, oysters, peppers, pork, sausages, soup, spinach) in relation to composition and sensory properties. Also covered are antimicrobial wine marinades, antioxidative and immunostimulating aspects, and adverse effects associated with wine consumption. The collated information and suggested research needs might facilitate and guide further studies needed to optimize the use of wines and byproducts to help improve microbial food safety and prevent or treat animal and human infections.

  10. Effects of glutamine on performance and intestinal mucosa morphometry of broiler chickens vaccinated against coccidiosis

    Directory of Open Access Journals (Sweden)

    Brenda Carla Luquetti

    2016-08-01

    Full Text Available ABSTRACT This study aimed to assess the effects of glutamine as feed additive on performance and intestinal mucosa morphometry of broiler chickens vaccinated against coccidiosis. A total of 400 day-old male chicks were randomly assigned to four treatments (NVNG – no vaccination, no glutamine supplementation; NVG – no vaccination, glutamine supplementation (10 g kg−1; VNG – vaccination, no glutamine supplementation; VG – vaccination, glutamine supplementation replicated four times with 25 birds per replicate. A commercial sprayed-on vaccine against coccidiosis containing Eimeria acervulina, E. maxima, E. mivati, and E. tenella was administered at the hatchery. Broiler performance was evaluated from 1-28 days, and morphometric parameters were analyzed at 14, 21, and 28 days of age. Body weight gain and feed intake were negatively affected by vaccination, but not by glutamine. Vaccination increased crypt depth in the duodenum and jejunum at 21 and 28 days. In conclusion, this study showed that glutamine was not able to increase weight gain of broiler chickens, irrespective of whether the animals were vaccinated or not against coccidiosis. Glutamine supplementation was able to improve feed conversion in vaccinated birds suggesting trophic effect on intestinal epithelium improving.

  11. Enhancement of resistance to coccidiosis and necrotic enteritis in broiler chickens by dietary muscadine pomace.

    Science.gov (United States)

    McDougald, L R; Hofacre, C; Mathis, G; Fuller, L; Hargrove, J L; Greenspan, P; Hartle, D K

    2008-12-01

    Muscadine pomace (MP), a by-product of the production of wine and juice from Vitis rotundifolia, was dried and tested in chickens for effects on primary resistance to coccidiosis, development of protective immunity after vaccination with live coccidia, and resistance to necrotic enteritis (NE) caused by the joint action of Clostridium perfringens and coccidia. To test primary resistance to coccidiosis, 2-wk-old chicks were given 2% or 5% MP in the diet and inoculated with Eimeria acervulina and E. maxima. Birds given MP at either level had significantly (P chickens were given 2% or 5% MP and grown to 42 days to test the palatability of MP. Birds given 2% MP in feed grew similarly to untreated controls, but birds given 5% had poorer average live weight. This suggested a negative effect on feed intake at the higher level. The effects of dietary 0.5% or 2.0% MP on immune protection were tested after live coccidiosis vaccination in the hatchery. Chicks were removed from each pen at 21 days of age and challenged with E acervulina, E. maxima, and E. tenella. Resistance to infection was improved by MP as suggested by significantly (P chickens. Chicks were inoculated with live coccidia at 14 days of age and dosed orally with live cultures of C perfringens on day 19, day 20, and day 21. Enteritis caused 48% mortality in the first study and 67% mortality in the second study. Dietary MP at 0.5-2.0% significantly (P chickens.

  12. Coccidia from bats (Chiroptera) of the world: a new Eimeria species in Pipistrellus javanicus from Japan.

    Science.gov (United States)

    Duszynski, D W

    1997-04-01

    Fecal samples from 56 Japanese bats representing 6 species in 2 families were examined for coccidian oocysts. Two of the 56 (Rhinolophidae), but only 2 sporulated oocysts were seen, which is not enough to describe a new species.

  13. Synergistic Effect of Dietary Curcuma, Capsicum, and Lentinus on enhancing local immunity against Eimeria acervulina infection

    Science.gov (United States)

    Traditionally, the application of prophylactic antibiotics has been successful in reducing infection-related morbidity and mortality in animal production. However, with increasing concerns over the widespread use of feed-added chemicals in animal production and the emergence of antibiotic resistant ...

  14. Down-regulation of chicken interleukin-17 receptor A in Eimeria infection

    Science.gov (United States)

    Both IL-17A and IL-17F are proinflammatory cytokines, which play an important role in intestinal homeostasis through their receptor signaling. In chickens, these two cytokines have been recently characterized, but to date, very little is known about their receptors and their functional activity. Th...

  15. Mitochondrial correlates of signaling processes involved with the cellular response to eimeria infection in broiler chickens

    Science.gov (United States)

    Host cellular responses to coccidiosis infection are consistent with elements of apoptosis, autophagy, and necrosis. These processes are enhanced in the cell through cell-directed signaling or repressed through parasite-derived inhibitors of these processes favoring the survival of the parasite. Acr...

  16. Protective effects of Aloe vera-based diets in Eimeria maxima-infected broiler chickens

    Science.gov (United States)

    Aloes have been widely used for a broad range of pharmacological activities, including parasitic problems. Avian coccidiosis is the most costly and wide-spread parasitic disease in the poultry industry, and has been mainly controlled by the use of chemotherapeutic agents. Due to the emergence of dru...

  17. Enterocyte protein tyrosine nitration in response to Eimeria infection in broilers

    Science.gov (United States)

    Activation of pathogen-sensing mechanisms in intestinal cells initiate the generation of pathway effectors that perturb normal nutritional enterocyte (ETC) functions. Among the conserved pathway mediator molecules generated are nitric oxide (NO) and superoxide anion (SOA) which are known to interac...

  18. Rodent malaria: BCG-induced protection and immunosuppression

    International Nuclear Information System (INIS)

    Smrkovski, L.L.; Strickland, G.T.

    1978-01-01

    One dose of 10 7 viable units of Mycobacterium bovis, strain BCG, protected a significant number of Swiss mice from a primary challenge with 10 4 thoracic sporozoites of Plasmodium berghei. Immunization with irradiated sporozoites induced greater protection than that observed in BCG-treated animals. Mice treated with BCG and surviving a primary sporozoite challenge were not protected from rechallenge, whereas mice immunized with irradiated sporozoites and surviving initial challenge of sporozoites were solidly immune to further challenge. Immunizing mice with BCG and irradiated sporozoites simulataneously resulted in a synergistic effect of increased protection against a primary challenge of sporozoites only if the two immunogens were administered on the same day and if the mice were challenged 1 to 3 days later. Mice given BCG and irradiated sporozoites and surviving a primary challenge of sporozoites were unable to survive rechallenge. BCG given to mice previously immunized with irradiated sporozoites suppressed their protective immunity against sporozoite challenge

  19. 21 CFR 520.2260a - Sulfamethazine oblet, tablet, and bolus.

    Science.gov (United States)

    2010-04-01

    ... and nonlactating dairy cattle. Treatment of bacterial pneumonia and bovine respiratory disease complex... mastitis (Streptococcus spp.), acute metritis (Streptococcus spp.), coccidiosis (Eimeria bovis and Eimeria...

  20. IgG2 antibodies against a clinical grade Plasmodium falciparum CSP vaccine antigen associate with protection against transgenic sporozoite challenge in mice.

    Directory of Open Access Journals (Sweden)

    Robert Schwenk

    Full Text Available The availability of a highly purified and well characterized circumsporozoite protein (CSP is essential to improve upon the partial success of recombinant CSP-based malaria vaccine candidates. Soluble, near full-length, Plasmodium falciparum CSP vaccine antigen (CS/D was produced in E. coli under bio-production conditions that comply with current Good Manufacturing Practices (cGMP. A mouse immunogenicity study was conducted using a stable oil-in-water emulsion (SE of CS/D in combination with the Toll-Like Receptor 4 (TLR4 agonist Glucopyranosyl Lipid A (GLA/SE, or one of two TLR7/8 agonists: R848 (un-conjugated or 3M-051 (covalently conjugated. Compared to Alum and SE, GLA/SE induced higher CS/D specific antibody response in Balb/c mice. Subclass analysis showed higher IgG2:IgG1 ratio of GLA/SE induced antibodies as compared to Alum and SE. TLR synergy was not observed when soluble R848 was mixed with GLA/SE. Antibody response of 3M051 formulations in Balb/c was similar to GLA/SE, except for the higher IgG2:IgG1 ratio and a trend towards higher T cell responses in 3M051 containing groups. However, no synergistic enhancement of antibody and T cell response was evident when 3M051 conjugate was mixed with GLA/SE. In C57Bl/6 mice, CS/D adjuvanted with 3M051/SE or GLA/SE induced higher CSP repeat specific titers compared to SE. While, 3M051 induced antibodies had high IgG2c:IgG1 ratio, GLA/SE promoted high levels of both IgG1 and IgG2c. GLA/SE also induced more potent T-cell responses compared to SE in two independent C57/BL6 vaccination studies, suggesting a balanced and productive T(H1/T(H2 response. GLA and 3M-051 similarly enhanced the protective efficacy of CS/D against challenge with a transgenic P. berghei parasite and most importantly, high levels of cytophilic IgG2 antibodies were associated with protection in this model. Our data indicated that the cGMP-grade, soluble CS/D antigen combined with the TLR4-containing adjuvant GLA/SE warrants further evaluation for protective responses in humans.

  1. Structural and evolutionary adaptation of rhoptry kinases and pseudokinases, a family of coccidian virulence factors

    Science.gov (United States)

    2013-01-01

    Background The widespread protozoan parasite Toxoplasma gondii interferes with host cell functions by exporting the contents of a unique apical organelle, the rhoptry. Among the mix of secreted proteins are an expanded, lineage-specific family of protein kinases termed rhoptry kinases (ROPKs), several of which have been shown to be key virulence factors, including the pseudokinase ROP5. The extent and details of the diversification of this protein family are poorly understood. Results In this study, we comprehensively catalogued the ROPK family in the genomes of Toxoplasma gondii, Neospora caninum and Eimeria tenella, as well as portions of the unfinished genome of Sarcocystis neurona, and classified the identified genes into 42 distinct subfamilies. We systematically compared the rhoptry kinase protein sequences and structures to each other and to the broader superfamily of eukaryotic protein kinases to study the patterns of diversification and neofunctionalization in the ROPK family and its subfamilies. We identified three ROPK sub-clades of particular interest: those bearing a structurally conserved N-terminal extension to the kinase domain (NTE), an E. tenella-specific expansion, and a basal cluster including ROP35 and BPK1 that we term ROPKL. Structural analysis in light of the solved structures ROP2, ROP5, ROP8 and in comparison to typical eukaryotic protein kinases revealed ROPK-specific conservation patterns in two key regions of the kinase domain, surrounding a ROPK-conserved insert in the kinase hinge region and a disulfide bridge in the kinase substrate-binding lobe. We also examined conservation patterns specific to the NTE-bearing clade. We discuss the possible functional consequences of each. Conclusions Our work sheds light on several important but previously unrecognized features shared among rhoptry kinases, as well as the essential differences between active and degenerate protein kinases. We identify the most distinctive ROPK-specific features

  2. Structural and evolutionary adaptation of rhoptry kinases and pseudokinases, a family of coccidian virulence factors.

    Science.gov (United States)

    Talevich, Eric; Kannan, Natarajan

    2013-06-06

    The widespread protozoan parasite Toxoplasma gondii interferes with host cell functions by exporting the contents of a unique apical organelle, the rhoptry. Among the mix of secreted proteins are an expanded, lineage-specific family of protein kinases termed rhoptry kinases (ROPKs), several of which have been shown to be key virulence factors, including the pseudokinase ROP5. The extent and details of the diversification of this protein family are poorly understood. In this study, we comprehensively catalogued the ROPK family in the genomes of Toxoplasma gondii, Neospora caninum and Eimeria tenella, as well as portions of the unfinished genome of Sarcocystis neurona, and classified the identified genes into 42 distinct subfamilies. We systematically compared the rhoptry kinase protein sequences and structures to each other and to the broader superfamily of eukaryotic protein kinases to study the patterns of diversification and neofunctionalization in the ROPK family and its subfamilies. We identified three ROPK sub-clades of particular interest: those bearing a structurally conserved N-terminal extension to the kinase domain (NTE), an E. tenella-specific expansion, and a basal cluster including ROP35 and BPK1 that we term ROPKL. Structural analysis in light of the solved structures ROP2, ROP5, ROP8 and in comparison to typical eukaryotic protein kinases revealed ROPK-specific conservation patterns in two key regions of the kinase domain, surrounding a ROPK-conserved insert in the kinase hinge region and a disulfide bridge in the kinase substrate-binding lobe. We also examined conservation patterns specific to the NTE-bearing clade. We discuss the possible functional consequences of each. Our work sheds light on several important but previously unrecognized features shared among rhoptry kinases, as well as the essential differences between active and degenerate protein kinases. We identify the most distinctive ROPK-specific features conserved across both active

  3. A new eimerian species (Apicomplexa: Eimeriidae) from the blue-fronted Amazon parrot Amazona aestiva L. (Aves: Psittacidae) in Brazil.

    Science.gov (United States)

    Hofstatter, P G; Guaraldo, A M A

    2011-12-01

    The Neotropical psittacine species Amazona aestiva, commonly known as the blue-fronted Amazon, is one of the most common and best-known psittacine birds kept as a pet worldwide. However, very little is known about the diseases or parasites of these birds. In this study, we describe a new species, Eimeria aestivae, associated with these parrots. The new species is characterized by: ovoid smooth oocysts (n  =  60), 36.8 (33.2-41.5) × 23.7 (21.7-25.7) µm, length/width ratio  =  1.55; polar granule present; ellipsoidal sporocysts (n  =  25), 19.8 (17.5-21.6) × 9.3 (8.3-9.9) µm; Stieda, sub-Stieda body, and sporocyst residuum present. Sporozoites (n  =  20), 2 per sporocyst, elongate and curved, 17.6 (15.8-19.2) × 3.8 (3.2-4.8) µm; each with 2 refractile bodies. The oocysts of the other 2 eimerian species described for Amazona are larger than those of the presented species, but they all seem to be closely related because of some similarities among them.

  4. Expanding the Therapeutic Spectrum of Artemisinin: Activity Against Infectious Diseases Beyond Malaria and Novel Pharmaceutical Developments

    Directory of Open Access Journals (Sweden)

    Thomas Efferth

    2016-08-01

    Full Text Available The interest of Western medicine in Traditional Chinese Medicine (TCM as a source of drug leads/new drugs to treat diseases without available efficient therapies has been dramatically augmented in the last decades by the extensive work and the outstanding findings achieved within this kind of medicine. The practice of TCM over thousands of years has equipped scientists with substantial experience with hundreds of plants that led to the discovery of artemisinin (qinghaosu, which is extracted from the medicinal plant Artemisia annua L. (qinghao. The unexpected success of artemisinin in combating malaria has drawn strong attention from the scientific community towards TCM. Artemisinin was discovered by Youyou Tu in 1972. Since then, several novel pharmacological activities based on the well-known properties of the sesquiterpene lactone structure with the oxepane ring and an endoperoxide bridge have been unravelled. Beyond malaria, artemisinin and its derivatives (artemisinins exert profound activities towards other protozoans (Leishmania, Trypanosoma, amoebas, Neospora caninum, and Eimeria tenella, trematodes (Schistosoma, liver flukes, and viruses (human cytomegalovirus, hepatitis B and C viruses. Less clear is the effect against bacteria and fungi. Based on the promising results of artemisinin and the first generation derivatives (artesunate, artemether, arteether, novel drug development strategies have been pursued. These included the synthesis of acetal- and non-acetal-type artemisinin dimeric molecules as well as developing nanotechnological approaches, e.g. artemisinin-based liposomes, niosomes, micelles, solid lipid nanocarriers, nanostructured lipid carriers, nanoparticles, fullerenes and nanotubes. The current review presents an overview on different aspects of artemisinins, including sources, chemistry, biological/pharmacological properties, types of infectious pathogens that are susceptible to artemisinins in vitro and in vivo, in

  5. Anticoccidial activity of the methanolic extract of Musa paradisiaca root in chickens.

    Science.gov (United States)

    Anosa, George Nnamdi; Okoro, O Josephine

    2011-01-01

    The study was designed to evaluate the anticoccidial activity of the methanolic extract of Musa paradisiaca root in chickens. The chickens were divided into six groups of 12 chickens each. Each chicken in five groups was infected with 8,000 infective coccidia (Eimeria tenella) oocysts at day 28 of age while one group served as uninfected control. At day 7 post-infection, two chickens remaining in each group were sacrificed for postmortem examination to confirm coccidiosis. Also at day 7 post-infection, each chicken in four infected groups was given graded doses (250, 500 and 1,000 mg/kg b.w.) of the extract or amprolium (conventional drug). Two groups (an infected and uninfected group) did not receive treatment. Parameters used to assess progress of infection and response to treatment included clinical signs typical of coccidiosis, oocyst count per gramme of faeces (OPG) and packed cell volume (PCV). Treatment of previously infected chickens with M. paradisiaca root extract resulted in a progressive decrease in severity of observed clinical signs, marked reductions in OPG and a gradual increase in PCV. In each case, the changes were dose dependent. There was no significant difference in mean OPG and mean PCV of the extract (at 1,000 mg/kg b.w.) and amprolium-treated groups at termination of the study (at day 50 of age). In the acute toxicity study, the extract was found to be non-toxic to the chickens even at the highest dose of 4,000 mg/kg b.w. The results of this study demonstrated that the extract has anticoccidial activity in a dose-dependent manner and at a dosage of 1,000 mg/kg b.w. had similar efficacy with amprolium in the treatment of chicken coccidiosis.

  6. Anticoccidial activity of Curcuma longa L. in broilers

    Directory of Open Access Journals (Sweden)

    Rao Zahid Abbas

    2010-02-01

    Full Text Available Comparative efficacy of turmeric (Curcuma longa L. crude powder and salinomycin sodium on the occurrence of coccidiosis and growth performance of broiler was evaluated. A total of 90, day-old chicks were randomly divided into six groups. From first day onward, ration was supplemented with 1, 2 and 3 % turmeric powder in groups 2, 3 and 4, respectively, group 1 received salinomycin sodium @ 12 g 50 kg-1 feed while groups 5 and 6 were kept as infected un-medicated and uninfected un-medicated controls. First five groups were infected with Eimeria tenella sporulated oocysts @ 1,00,000/chick at the age of 20 days. Body weight gain, feed consumption, feed conversion ratio were investigated throughout the experimental period, and bloody diarrhea and oocysts excretions were investigated at the first and the second week after infection. Maximum coccidiostatic effect was observed with turmeric (3% showing mild bloody diarrhea as compared to other infected groups receiving turmeric containing rations. This effect was comparable with a standard coccidiostat i.e., salinomycin sodium. Similarly, the weight gain in the groups treated with salinomycin sodium (2280g and 3% turmeric (2293g were also significantly higher (p < 0.05 than that of infected control group (1955g. In the groups treated with ration supplemented with 3% turmeric powder and salinomycin sodium, the peak excretion of oocysts was delayed about 1 or 2 days relative to the control infected group. Concentration-dependent coccidiostatic effect of turmeric suggested that further studies should be carried out to determine the possible maximum safe levels of turmeric with least toxic effects to be used as coccidiostat.

  7. Anticoccidial effect of Piper sarmentosum extracts in experimental coccidiosis in broiler chickens.

    Science.gov (United States)

    Wang, Dingfa; Zhou, Luli; Li, Wei; Zhou, Hanlin; Hou, Guanyu

    2016-06-01

    To study the anticoccidial effect of Piper sarmentosum extracts (PSE) in experimental broiler coccidiosis, 270 one-day-old Wenchang broiler chickens were randomly assigned to six groups, each with three replicates (n = 15). The six groups were blank control group (BC), negative control group (NC), positive control group (PC), and another three PSE addition groups. Chickens in three control groups were fed a basal diet without PSE supplementation. Chickens in the three PSE addition groups were fed a basal diet supplemented with PSE at 100 (T100), 200 (T200), and 300 (T300) mg/kg of feed, respectively. At 15 days of age, chickens in group NC, PC, and three PSE addition groups were challenged with an oral dose of 1 × 10(5) Eimeria tenella oocysts each chick. Chickens in group PC were fed with diclazuril solution in water for 5 days after 48 h with oocysts inoculation. The results showed that PSE and diclazuril improved growth performance and significantly (P < 0.05) decreased oocysts per gram in inoculated broiler chickens. PSE and diclazuril significantly (P < 0.05) decreased nitric oxide at 6 and 9 days post-inoculation relative to the NC group, respectively. At 6 and 9 days post-inoculation, PSE supplementation at 200 mg/kg in the diet increased concentration of interleukin 2 (IL-2) and interferon gamma (IFN-γ) (P < 0.05). PSE supplementation at 200 mg/kg in the diet significantly (P < 0.05) increased mRNA expressions of IFN-γ and IL-2 in the cecum of chickens at 9 days post-inoculation relative to the BC and NC group. The current results showed the anticoccidial properties, and beneficial effect on intestinal mucosa damage of PSE in broiler chickens that had been challenged by coccidiosis.

  8. A Model for Taxonomic Work on Homoxenous Coccidia: Redescription, Host Specificity, and Molecular Phylogeny of Eimeria ranae Dobell, 1909, with a Review of Anuran-Host Eimeria (Apicomplexa: Eimeriorina)

    Czech Academy of Sciences Publication Activity Database

    Jirků, Miloslav; Jirků, Milan; Oborník, Miroslav; Lukeš, Julius; Modrý, David

    2009-01-01

    Roč. 56, č. 1 (2009), s. 39-51 ISSN 1066-5234 R&D Projects: GA ČR GA206/03/1544 Institutional research plan: CEZ:AV0Z60220518 Keywords : Amphibia * Anura * experimental infections * metamorphosis * morphotypes * SSU rDNA sequence * tadpoles * ultrastructure Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.355, year: 2009

  9. Direct-fed microbials in broiler chickens challenged with Eimeria maxima or raised on Clostridium spp.-contaminated used litter

    Science.gov (United States)

    Dietary direct-fed microbials (DFMs) influence the composition of gut microbiota and enhance gut health in broiler chickens. Increasing scientific data have been gathered to show that gut microbiota plays an important role in the development of the immune system and the maintenance of homeostasis wi...

  10. 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...

  11. Temporal pattern changes in duodenal protein tyrosine nitration events in response to Eimeria acervulina infection in chickens

    Science.gov (United States)

    Intracellular generation of nitric oxide (NO) and superoxide anion (SOA) can result in the formation of 3'-nitrotyrosine proteins (NTp). Nitrated proteins usually are associated with significant perturbation in protein function, apoptosis, and cell death. We undertook the present study to establis...

  12. Estudo químico de cianobactérias marinhas e do cultivo misto entre a linhagem Geitlerinema sp CENA556 e o fungo Trichoderma atroviride, endófito da alga marinha Bostrychia tenella

    OpenAIRE

    Ezequiane Machado da Silva

    2016-01-01

    Organismos marinhos são reconhecidos como um rico reservatório de produtos naturais com estruturas moleculares excepcionais. Neste contexto, cianobactérias e fungos endofíticos emergiram como uma fonte promissora de substâncias bioativas. O objetivo deste trabalho foi buscar por substâncias biologicamente ativas a partir de linhagem de cianobactérias marinhas coletadas em Ubatuba, litoral do estado de São Paulo (Brasil) e explorar o potencial biossintético do fungo Trichoderma atroviride (end...

  13. Dietary protease can alleviate negative effects of a coccidiosis infection on production performance in broiler chickens

    NARCIS (Netherlands)

    Peek, H.W.; Klis, van der J.D.; Vermeulen, B.; Landman, W.J.M.

    2009-01-01

    Two experiments were conducted to determine the effect of dietary protease on coccidiosis infection, production performance, the intestinal mucus layer thickness, and brush border enzyme activity using broilers challenged with Eimeria spp. laboratory isolates (Eimeria acervulina, E. maxima and E.

  14. On the identity of the type species of the genus Telema (Araneae, Telemidae

    Directory of Open Access Journals (Sweden)

    Chunxia Wang

    2012-12-01

    Full Text Available Telema tenella Simon, 1882, the type species of genus Telema, is the only species of family Telemidae reported from Europe and all other 39 congeners occur far from it. However, it has never been properly described. In this paper T. tenella is redescribed and illustrated.

  15. Controlled human malaria infection by intramuscular and direct venous inoculation of cryopreserved Plasmodium falciparum sporozoites in malaria-naïve volunteers: effect of injection volume and dose on infectivity rates

    NARCIS (Netherlands)

    Gómez-Pérez, Gloria P.; Legarda, Almudena; Muñoz, Jose; Sim, B. Kim Lee; Ballester, María Rosa; Dobaño, Carlota; Moncunill, Gemma; Campo, Joseph J.; Cisteró, Pau; Jimenez, Alfons; Barrios, Diana; Mordmüller, Benjamin; Pardos, Josefina; Navarro, Mireia; Zita, Cecilia Justino; Nhamuave, Carlos Arlindo; García-Basteiro, Alberto L.; Sanz, Ariadna; Aldea, Marta; Manoj, Anita; Gunasekera, Anusha; Billingsley, Peter F.; Aponte, John J.; James, Eric R.; Guinovart, Caterina; Antonijoan, Rosa M.; Kremsner, Peter G.; Hoffman, Stephen L.; Alonso, Pedro L.

    2015-01-01

    Controlled human malaria infection (CHMI) by mosquito bite is a powerful tool for evaluation of vaccines and drugs against Plasmodium falciparum malaria. However, only a small number of research centres have the facilities required to perform such studies. CHMI by needle and syringe could help to

  16. Aspectos ultraestruturais do processo de divisão do Toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    Wanderley de Souza

    1974-02-01

    Full Text Available Neste trabalho é feita uma revisão sobre alguns aspectos biológicos do Toxoplasma gondii, principalmente sobre a ultraestrutura da forma interfásica e as modificações ultraestruturais que ocorrem no parasito durante o seu processo de divisão. Considera-se inicialmente o processo de divisão binária admitindo-se, porém, a possibilidade de que as imagens interpretadas como senão de divisão binária representem estágios da divisão por endodiogenia. Quanto à endodiogenia descrevem-se as alterações que ocorrem na "parasito mãe" durante o processo de formação dos dois "parasitos filhos". Este processo é semelhante no Toxoplasma gondii, Besnoitia jellisoni, Sarcocystis tenella e Frenkelia. Discute-se a possibilidade da formação de mais de dois "parasitos filhos" por um processo de endopoligenia, bem como o processo de esquizogonia. Os resultados mais recentes mostram que não existe esquizogonia nas formas vsgetativas do Toxoplasma gondii, senão que as imagens interpretadas como tal, ao microscópio ótico, são o resultado de endodiogenias sucessivas em que os endozoitas formados permanecem ligados entre si pela região posterior. A esquizogonia é, no entanto, encontrada nas formas que se desenvolvem no interior de células epiteliais do intestino do gato, que é o hospedeiro definitivo do Toxoplasma gondii. Discute-se o conceito de esquizogonia, comparando-o em três protozoários: Eimeria bovis, E. callospermophili e Plasmodium juxtanucleare, que apresentam diferenças entre si quanto ao processo de iniciação da individualização dos "parasitos filhos". Refere-se à recente hipótese que considera a endodiogenia como o processo fundamental de divisão dos esporozoárlos, ocorrendo na fase final da esquizogonia. Finalmente é acentuado o papel que a microscopia eletrônica aliada às modernas técnicas de citoquímica e imunocitoquimica poderá desempenhar no sentido de um melhor conhecimento da biologia do Toxoplasma