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Sample records for eimeria maxima recombinant

  1. Eimeria maxima microneme protein 2 delivered as DNA vaccine and recombinant protein induces immunity against experimental homogenous challenge.

    PubMed

    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 (P<0.05). The splenocytes from both vaccinated groups of chickens displayed significantly greater proliferation compared with the controls (P<0.05). Serum from chickens immunized with pVAX1-MIC2 and rEmMIC2 protein displayed significantly high levels of IL-2, IFN-γ, IL-10, IL-17, TGF-β and IL-4 (P<0.05) compared to those of negative controls. The challenge experiment results showed that both the recombinant protein and the DNA vaccine could obviously alleviate jejunum lesions, body weight loss, increase oocyst, decrease ratio and provide ACIs of more than 165. All the above results suggested that immunization with EmMIC2 was effective in imparting partial protection against E. maxima challenge and it could be an effective antigen candidate for the development of new vaccines against E. maxima.

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

    PubMed

    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.

  3. Conservation of proteins involved in oocyst wall formation in Eimeria maxima, Eimeria tenella and Eimeria acervulina.

    PubMed

    Belli, Sabina I; Ferguson, David J P; Katrib, Marilyn; Slapetova, Iveta; Mai, Kelly; Slapeta, Jan; Flowers, Sarah A; Miska, Kate B; Tomley, Fiona M; Shirley, Martin W; Wallach, Michael G; Smith, Nicholas C

    2009-08-01

    Vaccination with proteins from gametocytes of Eimeria maxima protects chickens, via transfer of maternal antibodies, against infection with several species of Eimeria. Antibodies to E. maxima gametocyte proteins recognise proteins in the wall forming bodies of macrogametocytes and oocyst walls of E. maxima, Eimeria tenella and Eimeria acervulina. Homologous genes for two major gametocyte proteins - GAM56 and GAM82 - were found in E. maxima, E. tenella and E. acervulina. Alignment of the predicted protein sequences of these genes reveals that, as well as sharing regions of tyrosine richness, strong homology exists in their amino-terminal regions, where protective antibodies bind. This study confirms the conservation of the roles of GAM56 and GAM82 in oocyst wall formation and shows that antibodies to gametocyte antigens of E. maxima cross-react with homologous proteins in other species, helping to explain cross-species maternal immunity.

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

    PubMed

    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.

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

    PubMed Central

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

    2017-01-01

    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. PMID:28432276

  6. Coccidian Merozoite Transcriptome Analysis From Eimeria Maxima In Comparison To Eimeria Tenella And Eimeria Acervulina

    USDA-ARS?s Scientific Manuscript database

    Using the Eimeria spp. population that infect chickens as a model for coccidian biology, we aimed to survey the transcriptome of E. maxima and contrast it to the two other Eimeria spp. for which transcriptome data are available, E. tenella and E. acervulina. Examining specifically the asexual intra...

  7. Eimeria maxima recombinant Gam82 gametocyte antigen vaccine protects against coccidiosis and augments humoral and cell-mediated immunity

    USDA-ARS?s Scientific Manuscript database

    Intestinal infection with Eimeria, the etiologic agent of avian coccidiosis, stimulates protective immunity to subsequent colonization by the homologous parasite, whilst cross-protection against heterologous species is poor. As a first step toward the development of a broad specificity Eimeria vacci...

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

    PubMed

    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.

  9. Coccidian merozoite transcriptome analysis from Eimeria maxima in comparison to Eimeria tenella and Eimeria acervulina.

    PubMed

    Schwarz, Ryan S; Fetterer, Raymond H; Rosenberg, George H; Miska, Katarzyna B

    2010-02-01

    With the Eimeria spp. populations that infect chickens used as a model for coccidian biology, we aimed to survey the transcriptome of Eimeria maxima and contrast it to the 2 other Eimeria spp. for which transcriptome data are available, i.e., Eimeria tenella and Eimeria acervulina . The asexual intracellular development stage, the merozoite, was specifically examined, and we used expressed sequence tag (EST) analysis to provide experimental evidence of transcription and a framework for understanding the merozoite stage of E. maxima . Of 2,680 individual ESTs obtained, 48.2% shared most significant (E < 10(-5)) homology to sequences from other apicomplexan species, primarily other Eimeria spp. and Toxoplasma gondii , and 47.5% were unique. Annotation of these ESTs enabled categorization to putative biological function and revealed an emphasis on translation, cytoskeleton, metabolism, signaling, transport, and protein folding, as well as the apicomplexan specific surface antigens and micronemes. Comparative analysis of abundantly expressed transcripts from merozoites of the 3 Eimeria spp. revealed a novel transcript common to all 3. Sharing no significant homology to any other sequence in public databases, this transcript was predicted to encode an Eimeria -specific protein (ESP) with 166-178 amino acids and 58.9-65.1% interspecific identity. A predicted signal peptide was identified, consistent with the assumption that ESP is a secreted protein. These annotated ESTs from E. maxima merozoites provide a resource for intra- and interspecific comparative analyses that will be useful in distinguishing the unique biology of coccidian parasites in relation to the diverse phylum of Apicomplexa.

  10. Different susceptibilities of Eimeria acervulina, Eimeria maxima, and Eimeria tenella oocysts to dessication

    USDA-ARS?s Scientific Manuscript database

    Outbreaks of avian coccidiosis may occur when susceptible chickens are raised on litter containing high concentrations of viable Eimeria oocysts. The purpose of this study was to compare the relative sensitivities of E. acervulina, E. maxima, and E. tenella oocysts to dessication. Sporulated E. ac...

  11. Characterization of the Eimeria maxima sporozoite surface protein IMP1

    USDA-ARS?s Scientific Manuscript database

    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...

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

    PubMed

    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.

  13. Characterization and localization of an Eimeria-specific protein in Eimeria maxima.

    PubMed

    Fetterer, Raymond H; Schwarz, Ryan S; Miska, Katarzyna B; Jenkins, Mark C; Barfield, Ruth C; Murphy, Charles

    2013-10-01

    A recently completed analysis of Eimeria maxima transcriptome identified a gene with homology to sequences expressed by E. tenella and E. acervulina but lacking homology with other organisms including other apicomplexans. This gene, designated Eimeria-specific protein (ESP), codes for a protein with a predicted molecular weight of 19 kDa. The ESP gene was cloned and the recombinant protein expressed in bacteria and purified for preparation of specific antisera. Quantitative RT-PCR showed transcription of ESP was low in unsporulated oocysts and after 24 h of sporulation. However, transcription nearly doubled after 48 h of sporulation and reached its highest levels in sporozoites (SZ) and merozoites (MZ). The protein was detectable by Western blot in both sporulated oocysts and in SZ and MZ. Immuno-localization by light microscopy identified ESP in paired structures in the anterior of SZ and MZ. Immuno-localization by electron microscopy identified ESP in MZ rhoptries but no specific staining of any SZ structures was detected. In addition, localization studies on intestinal sections recovered from birds 120-h post-infection indicates that oocysts do not stain with anti-ESP but staining of microgametocytes and developing oocysts was observed. The results indicate that ESP is associated with the rhoptry of E. maxima and that the protein may have functions in other developmental stages.

  14. Characterization and localization of an Eimeria-specific protein in Eimeria maxima

    USDA-ARS?s Scientific Manuscript database

    A recently completed analysis of Eimeria maxima transcriptome indentified a gene with homology to sequences expressed by E. tenella and E. acervulina but lacking homology with other organisms including other apicomplexans. This gene designated ESP codes for a protein with predicted molecular weight...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    PubMed

    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.

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

    PubMed Central

    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

  19. Induction of protective immunity against Eimeria tenella, Eimeria maxima, and Eimeria acervulina infections using dendritic cell-derived exosomes.

    PubMed

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

    2012-05-01

    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 T(h)1 cytokines, T(h)2 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 T(h)1 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 T(h)2 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.

  20. ANTIBODIES TO EIMERIA MAXIMA GAMETOCYTE ANTIGENS CROSS-REACT WITH EIMERIA TENELLA AND E. ACERVULINA: IMPLICATIONS FOR VACCINE DEVELOPMENT

    USDA-ARS?s Scientific Manuscript database

    It has been shown previously that vaccination with gamete specific molecules in Eimeria maxima offers protection via transfer of maternal antibodies (anti-EmAPGA), not just against infection with E. maxima, but also against E. tenella and E. acervulina. Antibodies to the gamete proteins recognise t...

  1. Differences in fecundity of Eimeria maxima strains exhibiting different levels of pathogenicity in its avian host

    USDA-ARS?s Scientific Manuscript database

    The purpose of this study was to determine the underlying cause of differences in pathogenicity of two Eimeria maxima strains (APU1 and APU2) observed during coccidiosis infection. At identical challenge doses, E. maxima APU1 always produces greater intestinal lesions and lower weight gain compared...

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

    PubMed

    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.

  3. Protective effect of hyperimmune egg yolk IgY antibodies against Eimeria tenella and Eimeria maxima infections.

    PubMed

    Lee, S H; Lillehoj, H S; Park, D W; Jang, S I; Morales, A; García, D; Lucio, E; Larios, R; Victoria, G; Marrufo, D; Lillehoj, Erik P

    2009-07-07

    Avian coccidiosis is caused by several distinct species of Eimeria protozoa and is the major parasitic disease of poultry of economic importance. As an alternative strategy to control avian coccidiosis without using prophylactic medications, we have investigated the efficacy of inducing passive immunity against coccidiosis by orally feeding hyperimmune IgY antibodies. In this study, a commercially available egg yolk powder, Supracox (SC), a purified IgY fraction of egg yolk prepared from hens hyperimmunized with three major species of Eimeria oocysts, were continuously fed to young chicks from hatch. Upon orally infecting these broiler chicks with Eimeria tenella and Eimeria maxima oocysts at 1 week of age, they showed significantly higher body weight gains (P<0.05) compared to the untreated controls. Furthermore, SC-fed chicks showed significantly less intestinal lesions and reduced fecal oocyst output compared to the untreated controls following oral infections with E. tenella and E. maxima. These results provide clear evidence that passive immunization of chicks with hyperimmune egg yolk IgY antibodies provide significant protection against E. tenella or E. maxima infections.

  4. Embryo vaccination of chickens using a novel adjuvant formulation stimulates protective immunity against Eimeria maxima infection.

    PubMed

    Lee, Sung-Hyen; Lillehoj, Hyun S; Jang, Seung I; Hong, Yeong-Ho; Min, Wongi; Lillehoj, Erik P; Yancey, Robert J; Dominowski, Paul

    2010-11-16

    Our previous study demonstrated that chickens immunized subcutaneously with an Eimeria recombinant profilin protein vaccine emulsified in a Quil A/cholesterol/DDA/Carbopol (QCDC) adjuvant developed partial protection against experimental avian coccidiosis compared with animals immunized with profilin alone. Because in ovo vaccination is presently used in commercial applications worldwide throughout the poultry industry, the current study was undertaken to investigate chicken embryo vaccination with profilin plus QCDC adjuvant. Eighteen day-old embryos were immunized with isotonic saline (control), profilin alone, QCDC alone, or profilin plus QCDC, and orally challenged with live Eimeria maxima at 7 days post-hatch. Body weight gain, fecal oocyst output, and intestinal cytokine transcript levels were assessed as measures of protective immunity. While immunization with profilin alone or QCDC alone did not alter body weight gain of infected chickens compared with the saline control group, vaccination with profilin plus QCDC increased body weight gain such that it was equal to the uninfected controls. Immunization with profilin plus QCDC also reduced fecal oocyst shedding compared with unimmunized controls, although in this case QCDC failed to provide an adjuvant effect since no difference was observed between the profilin-only and profilin/QCDC groups. Finally, increased levels of transcripts encoding IL-1β, IL-15, and IFN-γ were seen in the intestinal tissues of animals given profilin plus QCDC compared with the profilin-only or QCDC-only groups. In summary, this study demonstrates an adjuvant effect of QCDC on body weight gain and intestinal cytokine responses following in ovo vaccination of chickens with an Eimeria profilin vaccine. Published by Elsevier Ltd.

  5. Immune protection of microneme 7 (EmMIC7) against Eimeria maxima challenge in chickens.

    PubMed

    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 < .05). The splenocytes from both vaccinated groups of chickens displayed significantly greater proliferation response compared with the controls (P < .05). Serum from chickens immunized with pVAX1-MIC7 and rEmMIC7 displayed significantly high levels of interleukin-2, interferon-γ, IL-10, IL-17, tumour growth factor-β and IL-4 (P < .05) compared to those of negative controls. The challenge experiment results showed that both the recombinant antigen and the DNA vaccine could obviously alleviate jejunum lesions, body weight loss and enhance oocyst decrease ratio. The anti-coccidial index (ACI) of the pVAX1-MIC7 group was 167.84, higher than that of the recombinant MIC7 protein group, 167.10. Our data suggested that immunization with EmMIC7 was effective in imparting partial protection against E. maxima challenge in chickens and it could be an effective antigen candidate for the development of new vaccines against E. maxima.

  6. Embryo vaccination of chickens using a novel adjuvant formulation stimulates protective immunity against Eimeria maxima infection

    USDA-ARS?s Scientific Manuscript database

    Our previous study demonstrated that chickens immunized subcutaneously with an Eimeria recombinant profilin protein vaccine emulsified in a Quil A/cholesterol/DDA/Carbopol (QCDC) adjuvant developed partial protection against experimental avian coccidiosis compared with animals immunized with profili...

  7. Expression of nutrient transporters in duodenum, jejunum and ileum of Eimeria maxima- infected broiler chickens

    USDA-ARS?s Scientific Manuscript database

    The intracellular parasite, Eimeria maxima, invades epithelial cells of the intestine causing malabsorption, diarrhea, and decreased feed conversion resulting in significant economic losses to poultry producers. The uptake of amino acids is mediated by active transporters located on the basal and br...

  8. Nitrocellulose immunoblotting for identification and molecular gene cloning of Eimeria maxima antigens that stimulate lymphocyte proliferation.

    PubMed Central

    Bumstead, J M; Dunn, P P; Tomley, F M

    1995-01-01

    An immunoblotting technique was used to identify lymphostimulatory antigens within sized polypeptide fractions of Eimeria maxima sporozoites. Six fractions contained polypeptides that specifically stimulated the proliferation of immune lymphocytes in an in vitro assay, and polyclonal antisera were made in rabbits against these fractions. cDNA clones, isolated with antisera against a lymphostimulatory fraction of around 70 kDa, were found to encode four different antigens including a classical hsp70, a molecule homologous to an endoplasmic reticulum chaperonin (BiP/GRP), and a calcium-dependent serine/threonine protein kinase that appears homologous to a recently described molecule from Plasmodium falciparum. The protein kinase cDNA clone was overexpressed in Escherichia coli, and the recombinant antigen was found to induce both antibody and lymphoproliferative responses in chickens when administered subcutaneously. Thus, immunoblotting, in combination with in vitro lymphoproliferation assays, can be used as an initial screen for the identification of lymphostimulatory antigens from a complex pool of polypeptides, and a combination of cDNA cloning, expression, and immunization allows assessment of the lymphostimulatory activity of individual polypeptides. These studies should facilitate further evaluation of antigens that are potential candidates for inclusion in a recombinant vaccine against poultry coccidiosis. PMID:8548529

  9. Protective effect of hyperimmune egg yolk Ig Y antibodies against Eimeria tenella and Eimeria maxima infections

    USDA-ARS?s Scientific Manuscript database

    Avian coccidiosis is economically the most important parasitic disease of the poultry industry and is caused by intestinal infection with Eimeria spp. An alternative control strategy potentially applicable to intestinal diseases such as avian coccidiosis involves passive immunization using hyperimm...

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

    USDA-ARS?s Scientific Manuscript database

    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....

  11. Efficacy of a DNA Vaccine Carrying Eimeria maxima Gam56 Antigen Gene against Coccidiosis in Chickens

    PubMed Central

    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

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

    PubMed

    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 <0.1 distributed across five chromosomes. The highly significant SNPs were associated with body weight gain (three SNPs on GGA5, one SNP on GGA1 and one SNP on GGA3), plasma coloration measured as optical density at wavelengths in the range 465-510 nm (10 SNPs and all on GGA10) and the percentage of β2-globulin in blood plasma (15 SNPs on GGA1 and one SNP on GGA2). Biological pathways related to metabolic processes, cell proliferation, and primary innate immune processes were among the most frequent significantly enriched biological pathways. Furthermore, the network-based analysis produced two networks of high confidence, with one centered on large tumor suppressor kinase 1 (LATS1) and 2 (LATS2) and the second involving the myosin heavy chain 6 (MYH6). We identified several strong candidate genes and genomic regions associated with traits measured in

  13. Differences in fecundity of Eimeria maxima strains exhibiting different levels of pathogenicity in its avian host.

    PubMed

    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

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

    PubMed Central

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

    2011-01-01

    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. PMID:21998616

  15. Induction of protective immunity against Eimeria tenella, Eimeria maxima, and Eimeria acervulina infections using DC-derived exosomes

    USDA-ARS?s Scientific Manuscript database

    This study describes a novel immunization strategy against avian coccidiosis using exosomes derived from Eimeria parasite antigen (Ag)-loaded dendritic cells (DCs) in the absence of soluble Ag. Chicken intestinal DCs were isolated and pulsed in vitro with a mixture of sporozoite-extracted Ags from E...

  16. Large-scale investigation of the parameters in response to Eimeria maxima challenge in broilers.

    PubMed

    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 < 0.001), oocyst count (P < 0.05), plasma coloration for the optical density values between 450 and 490 nm (P < 0.001), albumin (P < 0.001), α1-globulin (P < 0.01), α2-globulin (P < 0.001), α3-globulin (P < 0.01), and β2-globulin (P < 0.001) were the most strongly affected parameters and expressed the greatest levels of variation. Plasma protein profiles proved to be a new, reliable parameter for measuring response to Eimeria maxima. Prediction of intestinal lesion score and fecal oocyst

  17. In vivo localization of antibodies raised against Eimeria maxima wall forming bodies during sexual intracellular development.

    PubMed

    Frölich, Sonja; Shahparee, Annisha; Wasinger, Valerie C; Wallach, Michael

    2014-11-01

    SUMMARY Apicomplexan parasites cause devastating diseases in humans and livestock. Previously we demonstrated that antibodies targeting transmissible forms of the apicomplexan parasite, Eimeria, are effective at reducing parasite shedding thus preventing the transmission of the disease. However, the mechanisms responsible have not been fully defined. Moreover, there is no direct evidence that the parasite-specific IgG antibodies can reach the parasite developing in the enterocytes of the infected chicken host. This study summarizes our efforts using host immunity, parasite proteomics and 3D microscopy to provide a step forward in our understanding of how this immune response works. Eimeria maxima is an important pathogen of poultry and used as a surrogate for a number of human pathogens including Toxoplasma and Plasmodium. Our studies demonstrate that immunization with the purified wall forming bodies (WFBs) results in a production of parasite-specific IgG antibodies, which have the ability to reach in situ gametocytes in the intestinal lumen and permeate the enterocyte/parasite membranes in order to bind to the cytoplasmic Type 1 and Type 2 WFBs. This raises the intriguing possibility that via this process antibodies block the development of Eimeria maxima in vivo.

  18. Cross immunity of DNA vaccine pVAX1-cSZ2-IL-2 to Eimeria tenella, E. necatrix and E. maxima.

    PubMed

    Shah, Mohammed Ali A; Xu, Lixin; Yan, Ruofeng; Song, Xiaokai; Li, Xiangrui

    2010-03-01

    The study describes cross protection experiments with chimeric DNA vaccine pVAX1-cSZ2-IL-2 to determine its efficacy against four important Eimeria species. Seven-day-old chickens were randomly divided into nine groups; group 1 negative control, groups 2, 3, 4, 5 positive controls; and groups 6, 7, 8 and 9 experimental groups. On days 7 and 14, groups 1-5 were injected with TE buffer, and groups 6-9 with the vaccine. At 21 days of age, all chickens were inoculated with 5 x 10(4) sporulated oocysts except for the negative control. Groups 2 and 6 were inoculated with Eimeria tenella, groups 3 and 7 with Eimerianecatrix, groups 4 and 8 with Eimeria acervulina and groups 5 and 9 with Eimeria maxima. Seven days later, all chickens were weighed and slaughtered to obtain intestinal samples. Efficacy of immunization was evaluated on the basis of oocyst decrease ratio, lesion score, body-weight gain and anti-coccidial index. The results indicated that the recombinant plasmid can induce host immune responses by alleviating intestinal lesions, body weight loss and oocyst ratio and imparting good protection against E. tenella and E.acervulina, medium protection against E. necatrix but little effect against E. maxima. It is concluded that the conserved antigen can provide cross protection and should be explored further.

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

    PubMed Central

    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

  20. Protection against coccidiosis in outbred chickens elicited by gamma-irradiated Eimeria maxima.

    PubMed

    Jenkins, M C; Chute, M B; Danforth, H D

    1997-01-01

    In an effort to develop an attenuated coccidiosis vaccine against coccidiosis, we exposed Eimeria maxima oocysts to an optimum dose of gamma irradiation (17 kRad) that does not affect sporozoite invasion of the intestinal mucosa but does prevent asexual parasite development. Irradiated E. maxima oocysts were suspended in gelatin slabs and placed in battery cages for ingestion by 1-day-old chickens. Separate groups of chickens were given gelatin slabs containing nonirradiated E. maxima oocysts or were inoculated per os with either irradiated or nonirradiated E. maxima oocysts. Chickens infected with irradiated or nonirradiated oocysts by either oral inoculation or gel delivery showed a dose-dependent protection against weight loss associated with E. maxima challenge compared with unimmunized controls. In general, nonirradiated oocysts elicited protective immunity at lower immunization doses compared with irradiated oocysts. These experiments were extended to a floor pen study wherein 1-day-old male and female broiler chickens were given irradiated or nonirradiated E. maxima oocysts in gelatin slabs in hatching boxes and challenged at 4 wk of age. A significant reduction (P < 0.05) in lesion scores was observed for chickens immunized with either irradiated or nonirradiated oocysts compared with unimmunized controls. Although no significant difference (P > 0.05) was observed in weight gain between these groups, both male and female chickens inoculated with irradiated E. maxima oocysts showed about a 10% greater weight gain than unimmunized controls. For both male and female chickens, average weights at challenge were greater in groups that were immunized with 17-kRad-irradiated E. maxima oocysts compared with those animals immunized with nonirradiated oocysts.

  1. Comparative microarray analysis of intestinal lymphocytes following Eimeria acervulina, E. maxima, or E. tenella infection in the chicken

    USDA-ARS?s Scientific Manuscript database

    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 ...

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

    PubMed Central

    Frölich, Sonja; Wallach, Michael

    2016-01-01

    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. PMID:27352801

  3. Protection of Broiler Chicks Housed with Immunized Cohorts Against Infection with Eimeria maxima and E. acervulina.

    PubMed

    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

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

    PubMed

    Frölich, Sonja; Wallach, Michael

    2015-06-01

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

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

    PubMed

    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.

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

    PubMed

    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 < 0.0001). Lastly, the interaction between isolate and gross lesion scores (P

  7. Effects of Natustat supplementation on performance, feed efficiency and intestinal lesion scores in broiler chickens challenged with Eimeria acervulina, Eimeria maxima and Eimeria tenella.

    PubMed

    Duffy, C F; Mathis, G F; Power, R F

    2005-06-30

    The effects of dietary supplementation of Natustat, a proprietary plant derived product (Alltech Inc., KY, USA) and Salinomycin, on performance, feed efficiency and intestinal lesion scores were observed during two Eimeria challenge trials in broiler chickens. In the first trial chickens were challenged with Eimeria sp. via infecting the litter with a known amount of Eimeria oocysts. In the second trial the source of the Eimeria challenge was the litter from the first trial and the same treatment groups were assigned to the same pens as in the initial trial. Birds were placed 55 per pen with seven pens per treatment. Performance parameters were recorded on days 21 and 42 during both trials. Intestinal lesion scores were assessed on days 14 and 21 during Trial 1 and on day 21 during Trial 2. Average weight gain and feed conversion ratios were significantly improved in the Natustat and Salinomycin treatment groups when compared to the non-supplemented infected group. Furthermore, lesion scores were lower on all sampling days in the Natustat and Salinomycin groups when compared to the non-supplemented group. However, only lesions associated with Eimeria tenella were significantly lowered by Natustat and Salinomycin supplementation. Natustat and Salinomycin were equivalent in alleviating the negative performance effects associated with coccidiosis challenge. In summary, Natustat has the potential to be used as a natural alternative to chemotherapeutic drugs for Eimeria control.

  8. Comparative microarray analysis of intestinal lymphocytes following Eimeria acervulina, E. maxima, or E. tenella infection in the chicken.

    PubMed

    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.

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

    PubMed Central

    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

  10. 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.

    PubMed

    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

  11. Development by genetic recombination of a line of Eimeria tenella resistant to robenidine, decoquinate and amprolium.

    PubMed

    Chapman, H D

    1984-01-01

    A line of Eimeria tenella (H) resistant to robenidine, decoquinate and amprolium has been produced by genetic recombination. It was not possible to obtain a cross between this line and lines resistant to clopidol or arprinocid and halofuginone. Recombination between lines resistant to arprinocid and halofuginone resulted in a loss of pathogenicity.

  12. Analysis of differentially expressed genes in the precocious line of Eimeria maxima and its parent strain using suppression subtractive hybridization and cDNA microarrays.

    PubMed

    Dong, Hui; Lin, Jiaojiao; Han, Hongyu; Jiang, Lianlian; Zhao, Qiping; Zhu, Shunhai; Huang, Bing

    2011-04-01

    The precocious line of Eimeria spp., obtained by repeated passages of oocysts initially collected from feces of previously infected chickens, has unique phenotypes and plays an important role in immunizing chickens against coccidiosis. However, the genetic basis of precocious phenotype in Eimeria is still poorly understood. To investigate gene expression changes in sporulated oocysts between the precocious line of E. maxima and its parent strain, subtractive cDNA libraries were constructed by suppression subtractive hybridization (SSH). A total of 3,164 cDNA fragments were selected from the SSH cDNA libraries to fabricate cDNA microarrays and further identify the differentially expressed genes. The credibility of the microarray data was verified by real-time PCR. A total of 360 valid expressed sequence tags (ESTs) were obtained, which represented 32 unique sequences. Twenty-one genes were validated as downregulated and 11 genes as upregulated in the precocious line. Homology searching of the public sequence database showed that six genes encoded proteins homologous with previously reported proteins, including rhomboid-like protein and transhydrogenase of E. tenella, serpin, and cation-transporting ATPase of E. acervulina, a heat-shock protein of E. maxima, and a conserved hypothetical protein of Toxoplasma gondii. Thus, the remaining 26 ESTs have not been previously reported. Further characterization of these differentially expressed genes will be useful in understanding the genetic basis for the precocious phenotype in Eimeria spp.

  13. Ultrastructural evaluation of the effects of diclazuril on the endogenous stages of Eimeria maxima and E. brunetti in experimentally inoculated chickens.

    PubMed

    Verheyen, A; Maes, L; Coussement, W; Vanparijs, O; Lauwers, F; Vlaminckx, E; Marsboom, R

    1989-01-01

    The ultrastructural morphology of the different endogenous stages of Eimeria maxima and E. brunetti was evaluated after oral treatment of inoculated chickens with a single dose of 5 mg/kg diclazuril. The drug induced no ultrastructural change in the growth or differentiation of the various schizont stages of both Eimeria spp. In E. maxima, the micromorphological appearance of micro- and macrogamonts developing from the blast from to maturation also remained unaffected by drug treatment. However, in all fertilized macrogamonts the normal pattern of oocyst wall establishment was completely disturbed, resulting in the formation of an abnormally thickened, incomplete oocyst wall and the necrosis of the zygote. In E. brunetti, the growth and nuclear division during microgametogenesis were not affected but differentiation was clearly abnormal. In comparison with the controls, this abnormal differentiation was characterized by a less extensive enlargement of the parasite surface area, aberrant morphological configurations of condensed heterochromatin, intracytoplasmic flagella formation, and glycogen accumulation. Finally, the complete degeneration of all microgamonts ensued. The growth and differentiation leading to mature macrogamonts was not disturbed; however, subsequent oocyst wall formation was largely precluded and the macrogamonts proceeded to degenerate completely. We conclude that diclazuril treatment primarily affected particular stages in the sexual development of both Eimeria spp., resulting in the complete eradication of these coccidian species.

  14. A recombinant Eimeria protein inducing interferon-gamma production: comparison of different gene expression systems and immunization strategies for vaccination against coccidiosis.

    PubMed

    Lillehoj, H S; Choi, K D; Jenkins, M C; Vakharia, V N; Song, K D; Han, J Y; Lillehoj, E P

    2000-01-01

    A rabbit antiserum against an 18- to 27-kD native protein fraction (F3) from Eimeria acervulina merozoites identified a cDNA (3-1E) containing a 1086-base pair insertion with an open reading frame of 170 amino acids (predicted molecular weight, 18,523). The recombinant 3-1E cDNA expressed in Escherichia coli produced a 60-kD fusion protein and a 23-kD protein after factor Xa treatment of the fusion protein. Both proteins were reactive with the F3 antiserum by western blot analysis. A rabbit antiserum against a synthetic peptide deduced from the amino acid sequence of the 3-1E cDNA reacted with a 27-kD recombinant 3-1E protein expressed in Sf9 insect cells and a 20-kD native protein expressed by E. acervulina sporozoites and Eimeria tenella sporozoites and merozoites. By immunofluorescence staining, a monoclonal antibody produced against the recombinant 3-1E protein reacted with sporozoites and merozoites of E. acervulina, E. tenella, and Eimeria maxima. Spleen lymphocytes from E. acervulina-immune chickens showed antigen-specific proliferation and interferon (IFN)-gamma production upon stimulation with the recombinant 3-1E protein, indicating that the protein activates cell-mediated immunity during coccidiosis. Immunization of chickens with either the E. coli- or Sf9-expressed recombinant 3-1E protein with adjuvant, or direct injection of the 3-1E cDNA, induced protective immunity against live E. acervulina. Simultaneous injection of the recombinant 3-1E protein, or the 3-1E cDNA, with cDNAs encoding chicken IFN-gamma or interleukin (IL)-2/15 further enhanced protective immunity. These results indicate that the recombinant E. acervulina 3-1E cDNA or its polypeptide product may prove useful as vaccines against avian coccidiosis.

  15. Immunoenhancing effects of MontanideTM ISA oil-based adjuvants on recombinant coccidia antigen vaccination against Eimeria acervulina infection

    USDA-ARS?s Scientific Manuscript database

    The current study was conducted to investigate the immunoenhancing effects of Montanide' adjuvants on protein subunit vaccination against avian coccidiosis. Broiler chickens were immunized subcutaneously with a purified Eimeria acervulina recombinant profilin protein, either alone or mixed with one ...

  16. The anticoccidial effects of amprolium, dinitolmide and monensin against Eimeria maxima, E, brunetti and E. acervulina with particular reference to oocyst sporulation.

    PubMed

    Joyner, L P; Norton, C C

    1977-10-01

    Chicks infected with the Weybridge strains of Eimeria maxima and E. acervulina were not protected by the normal levels of amprolium, but the sporulation of the oocysts was inhibited. With higher drug concentrations, fewer oocysts were produced. E brunetti was not so markedly affected, although oocyst sporulation was reduced by the higher dosage levels. The effects were not enhanced by the inclusion of ethopabate. With dinitolmide the phenomenon was not so marked, although oocysts of E. maxima and E. acervulina were reduced in numbers by the normal drug concentration and sporulation was reduced when this level was increased. Much higher drug levels were required to obtain these effects with E. brunetti. Monensin at 120 ppm affected neither oocyst numbers nor their sporulation in any of the species tested. The significance of the effects of anticoccidial drugs on gametogony and sporogony is discussed.

  17. Effects of feed-borne Fusarium mycotoxins and an organic mycotoxin adsorbent on immune cell dynamics in the jejunum of chickens infected with Eimeria maxima.

    PubMed

    Girgis, George N; Barta, John R; Girish, Channarayapatna K; Karrow, Niel A; Boermans, Herman J; Smith, Trevor K

    2010-12-01

    An experiment was conducted to explore the effects of Fusarium mycotoxins, common animal feed contaminants, on intestinal immune responses to coccidia (Eimeria) in chickens. Effects of feed-borne Fusarium mycotoxins and a polymeric glucomannan mycotoxin adsorbent (GMA) on immune cell populations were studied in the jejunum of broiler breeder pullets using an Eimeria maxima infection model. Birds were fed a control diet, a diet naturally contaminated with Fusarium mycotoxins, contaminated diet plus 0.2% GMA, or control diet plus 0.2% GMA. Contaminated diets contained up to 6.5μg/g deoxynivalenol (DON), 0.47μg/g 15-acetyl-DON and 0.73μg/g zearalenone. Birds received a primary oral inoculation (1000 oocysts/bird) with E. maxima USDA strain 68 at 2 weeks of age and a secondary oral inoculation (30,000 oocysts/bird) with the same strain at 4 weeks of age. Diet-related differences in CD4(+) cell, CD8(+) cell and macrophage recruitment pattern into the jejunum were observed following both the primary and secondary infections. It was concluded that feed-borne Fusarium mycotoxins and GMA have the potential to modulate immune response to coccidial infections.

  18. Recombinant chicken interferon-gamma-mediated inhibition of Eimeria tenella development in vitro and reduction of oocyst production and body weight loss following Eimeria acervulina challenge infection.

    PubMed

    Lillehoj, H S; Choi, K D

    1998-01-01

    Recombinant chicken interferon-gamma (chIFN-gamma) was produced in CHO-K1 or Spodoptera frugiperda (SF9) insect cells by transfection with a pcDNA vector or recombinant baculovirus (SF9-interferon-gamma [IFN-gamma] carrying the chIFN-gamma gene. A rabbit antibody against a synthetic peptide corresponding to an immunogenic portion of chIFN-gamma recognized a 22-23-kDa band in SF9-IFN-gamma cell extracts by western blot analysis. Biological activity of recombinant chIFN-gamma was shown by its inhibition of vesicular stomatitis virus-induced cytotoxicity of chicken embryonic fibroblast cells in vitro. To investigate the role of chIFN-gamma during Eimeria infection, CHCC-OU2 chicken cells either pretreated with chIFN-gamma or stably transfected with the chIFN-gamma gene were infected with Eimeria tenella sporozoites. IFN-gamma demonstrated significant reductions in intracellular sporozoite development without affecting sporozoite invasion of host cells. Furthermore, chickens treated with recombinant chIFN-gamma showed decreased oocyst production and significant improvement in body weight gain following Eimeria acervulina challenge infection. These results provide the first direct evidence that chIFN-gamma exerts an inhibitory effect against Eimeria and provides a rational basis for use of this cytokine as a vaccine adjuvant against coccidiosis.

  19. Analysis of differentially expressed genes in two immunologically distinct strains of Eimeria maxima using suppression subtractive hybridization and dot-blot hybridization

    PubMed Central

    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

  20. Embryo vaccination against Eimeria tenella and E. acervulina infections using recombinant proteins and cytokine adjuvants.

    PubMed

    Lillehoj, Hyun S; Ding, Xicheng; Dalloul, Rami A; Sato, Takanori; Yasuda, Atsushi; Lillehoj, Erik P

    2005-06-01

    Avian coccidiosis is an intestinal disease caused by protozoa of the genus Eimeria. To investigate the potential of recombinant protein vaccines to control coccidiosis, we cloned 2 Eimeria sp. genes (EtMIC2 and 3-1E), expressed and purified their encoded proteins, and determined the efficacy of in ovo immunization to protect against Eimeria infections. Immunogen-specific serum antibody titers, parasite fecal shedding, and body weight gains were measured as parameters of disease. When administered alone, the recombinant EtMIC2 gene product induced significantly higher antibody responses, lower oocyst fecal shedding, and increased weight gains compared with nonvaccinated controls following infection with E. tenella. Combined embryo immunization with the EtMIC2 protein plus chicken cytokine or chemokine genes demonstrated that all 3 parameters of vaccination were improved compared with those of EtMIC2 alone. In particular, covaccination with EtMIC2 plus interleukin (IL)-8, IL-16, transforming growth factor-beta4, or lymphotactin significantly decreased oocyst shedding and improved weight gains beyond those achieved by EtMIC2 alone. Finally, individual vaccination with either EtMIC2 or 3-1E stimulated protection against infection by the heterologous parasite E. acervulina. Taken together, these results indicate that in ovo vaccination with the EtMIC2 protein plus cytokine/chemokine genes may be an effective method to control coccidiosis.

  1. Eimeria tenella, E. acervulina and E. maxima: studies on the development of resistance to diclazuril and other anticoccidial drugs in the chicken.

    PubMed

    Chapman, H D

    1989-10-01

    Resistance to diclazuril was induced by 10 passages of the Houghton strains of Eimeria acervulina and E. tenella in chickens given progressively greater concentrations of the drug. This resistance was, however, not complete since the drug retained some efficacy against the drug-passaged lines. Attempts to passage the Houghton strain of E. maxima in birds medicated with concentrations of diclazuril greater than 0.016 parts per million (ppm) were unsuccessful and after 10 passages at this concentration resistance had not developed. Resistance to methyl benzoquate developed after 6 passages of E. tenella in medicated chickens but resistance to amprolium was only partial after 10 passages. The pathogenicity of diclazuril- and amprolium-resistant lines of E. tenella was lower than that of the parental line. Diclazuril was effective against lines of E. tenella resistant to amprolium, arprinocid, clopidol, dinitolmide, halofuginone, methyl benzoquate, monensin and robenidine.

  2. In ovo administration of CpG oligodeoxynucleotides and the recombinant microneme protein MIC2 protects against Eimeria infections.

    PubMed

    Dalloul, Rami A; Lillehoj, Hyun S; Klinman, Dennis M; Ding, Xicheng; Min, Wongi; Heckert, Robert A; Lillehoj, Erik P

    2005-05-02

    We have previously demonstrated that short oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODNs) exert a positive effect on weight loss and oocyst shedding associated with Eimeria infection when injected in vivo. The present work investigated the effects of in ovo vaccination with CpG ODNs and an Eimeria recombinant microneme protein (MIC2), alone or in combination, on susceptibility to coccidiosis. In ovo injection of CpG ODNs alone enhanced resistance to experimental Eimeria acervulina infection as best exemplified by reduced oocyst shedding. Two CpG ODNs reduced the oocyst load, but did not affect weight gain. When co-administered with the recombinant microneme protein, both ODNs reduced oocyst shedding; however, only ODN D19 plus MIC2 consistently improved weight gain. Vaccinating with ODN 2006 or MIC2 protein curtailed oocyst shedding but did not enhance weight gain in Eimeria tenella-infected birds. Co-administration of CpG ODN and MIC2 did not have an additive effect in reducing the oocyst output; however, it resulted in the highest and lowest Ab response before and after Eimeria tenella infection, respectively. Collectively, CpG ODNs administered in ovo demonstrated immunoenhancing and adjuvant effects following Eimeria infections.

  3. Genetic recombination of precociousness and anticoccidial drug resistance in Eimeria tenella.

    PubMed

    Jeffers, T K

    1976-10-12

    Two strains of Eimeria tenella differing in decoquinate-resistance and developmental rate were crossed. Strain Wis-F was decoquinate-sensitive (DS) and precocious (P+), while strain 368 was decoquinate-resistant (DR) and had a normal developmental rate (P-). Cultures of the parent strains and a culture derived from a mixture of parent strain oocysts were propagated through drug and developmental barriers to select parasites with the respective parental phenotypes (DS/P + and DR/P-) and the recombinant phenotype (DR/P+). The ability of a portion of the population in the strain-cross culture to reproduce in the presence of simultaneously imposed drug and developmental barrier showed that they were the recombinant phenotype (DR/P+), which had been produced through fertilization by gametes of opposite parent strains of E. tenella.

  4. Immunogenicity of recombinant attenuated Salmonella enterica serovar Typhimurium vaccine strains carrying a gene that encodes Eimeria tenella antigen S07

    USDA-ARS?s Scientific Manuscript database

    In an attempt to develop an efficacious vaccine against avian coccidiosis, research was conducted using Type III Secretion System (TTSS) of Salmonella to deliver Eimeria antigens into the cytoplasm of host cells. Once delivered, recombinant protein may enter the MHC I antigen processing pathway for...

  5. Relative disease susceptibility and clostridial toxin antibody responses in three commercial broiler lines coinfected with Clostridium perfringens and Eimeria maxima using an experimental model of necrotic enteritis.

    PubMed

    Jang, Seung I; Lillehoj, Hyun S; Lee, Sung-Hyen; Lee, Kyung Woo; Lillehoj, Erik P; Hong, Yeong Ho; An, Dong-Jun; Jeoung, D Hye-Young; Chun, Ji-Eun

    2013-09-01

    Necrotic enteritis is an enteric disease of poultry resulting from infection by Clostridium perfringens with coinfection 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 enteritis. Day-old male Cobb, Ross, and Hubbard broilers were orally infected with viable C. perfringens and E. maxima and fed a high-protein diet to promote the development of experimental disease. Body weight loss, intestinal lesions, and serum antibody levels against alpha-toxin and necrotic enteritis B-like (NetB) toxin were measured as parameters of disease susceptibility and host immune response. Cobb chickens exhibited increased body weight loss compared with Ross and Hubbard breeds and greater gut lesion severity compared with Ross chickens. NetB antibody levels were greater in Cobb chickens compared with the Ross or Hubbard groups. These results suggest that Cobb chickens may be more susceptible to necrotic enteritis in the field compared with the Ross and Hubbard lines.

  6. A comparison of sporozoite transport after homologous and heterologous challenge in chickens immunized with the Guelph strain or the Florida strain of Eimeria maxima.

    PubMed

    Beattie, S E; Fernando, M A; Barta, J R

    2001-02-01

    The two strains of Eimeria maxima, Guelph and Florida, used in this study were previously shown to only partially cross-protect immunologically with respect to lesion scores, weight gains and feed conversions after heterologous challenge. In this paper, we provide evidence that this partial lack of cross-protection is manifested at the level of sporozoite transport. In birds immunized and challenged with the homologous strain, sporozoites accumulated in the lamina propria and were blocked from further movement into the crypts by 72 h post-challenge, unlike the situation observed in naive birds. Fewer than 5% of sporozoites were found in the crypts by 72 h post-challenge. In immunized birds challenged with the heterologous strain, fewer sporozoites reached the crypts than in naive birds but at least four times as many sporozoites successfully migrated to the crypts, when compared with birds challenged with the homologous strain. The degree of cross-protection afforded by the heterologous strain as measured by sporozoite transport success was not equally reciprocal.

  7. Recombinant Eimeria protozoan protein elicits resistance to acute phlebovirus infection in mice but not hamsters.

    PubMed

    Gowen, Brian B; Smee, Donald F; Wong, Min-Hui; Judge, John W; Jung, Kie-Hoon; Bailey, Kevin W; Pace, Anne M; Rosenberg, Barnett; Sidwell, Robert W

    2006-06-01

    A protein antigen from an Eimeria protozoan has recently been reported to induce antitumor activity in mice. This activity most likely results from the strong induction of interkeukin-12 (IL-12) and gamma interferon (IFN-gamma), which are also essential factors in the establishment of protective immunity against viral infection. We evaluated recombinant Eimeria antigen (rEA) as a potential immunotherapeutic agent in mouse and hamster models of acute phleboviral disease. Punta Toro virus (PTV) was highly sensitive to a single dose of nanogram quantities of rEA in the mouse infection model. Intraperitoneal treatment with rEA also reduced virus load and liver damage associated with PTV infection. IL-12 was elicited following exposure of uninfected mice to quantities of rEA of 10 ng or greater, and the levels peaked at between 3 and 8 h postexposure. IFN-gamma release was induced more slowly and required less rEA (1 ng) to produce a significant rise in systemic levels. The induction of IL-12 and IFN-gamma involved in the coordination of innate and adaptive immune responses to microbial pathogens required myeloid differentiation factor 88, a signaling adaptor shared by most members of the Toll-like receptor (TLR) family. Despite encouraging results in the murine system, rEA failed to protect hamsters challenged with PTV. Our findings suggest that hamsters may lack functional TLR11, which has recently been shown to recognize a profilin-like protein homologous to rEA from the protozoan Toxoplasma gondii. Further investigation into the immunostimulatory capacity of rEA in other mammalian systems is necessary.

  8. Partial protective of chickens against Eimeria tenella challenge with recombinant EtMIC-1 antigen.

    PubMed

    Qi, N S; Wang, Y Y; Liao, S Q; Wu, C Y; Lv, M N; Li, J; Tong, Z X; Sun, M F

    2013-06-01

    Eimeria tenella microneme protein 1 (EtMIC-1) is highly conserved with TgMIC-2, which is involved in parasite binding specifically to host cells. Little is known about the immune responses and protective efficacy against E. tenella infection with EtMIC-1 antigen. In the present study, the recombinant proteins of E. tenella mature MIC-1 and adhesive domain (von Willebrand factor type A domain, EtMIC-1-VD) were obtained, protective efficacy against E.tenella infection and the mucosal immune response, which is induced in broilers was evaluated. The antibody levels and the transcription profiles of cytokine of chickens, such as interleukin-12 (IL-12) and interferon-γ (IFN-γ), were detected after being immunized three times with the recombinant EtMIC-1 and EtMIC-1-VD by ELISA assay and quantitative real-time PCR, respectively. The results showed that both groups of chickens, after being immunized with 100 μg EtMIC-1 or EtMIC-1-VD antigen, induced about tenfold higher IgG levels compared to the nonimmune groups. The transcription profiles of IL-12 and IFN-γ of the immunized groups were significantly higher than the control groups as well. The anticoccidial index of the group immunized with 100 μg EtMIC-1 and the group immunized with 100 μg EtMIC-1-VD were 167.2 and 165.5, respectively, which are significantly higher than low-dose immunized groups and challenged control groups. Our data suggests that VD domain is the key functional structure of EtMIC-1 that could trigger a significant humoral and cellular response against E. tenella infection, and EtMIC-1 had the potential in imparting partial protection in chickens against homologous challenge.

  9. Strengthened tumor antigen immune recognition by inclusion of a recombinant Eimeria antigen in therapeutic cancer vaccination.

    PubMed

    Quiroga, Dionisia; Aldhamen, Yasser A; Appledorn, Daniel M; Godbehere, Sarah; Amalfitano, Andrea

    2015-04-01

    The need for novel, effective adjuvants that are capable of eliciting stronger cellular and humoral adaptive immune responses to antigenic targets is well understood in the vaccine development field. Unfortunately, many adjuvants investigated thus far are either too toxic for human application or too weak to induce a substantial response against difficult antigens, such as tumor-associated antigens (TAAs). In spite of this trend, clinical investigations of recombinant Eimeria antigen (rEA) have revealed this protein to be a non-toxic immunogenic agent with the ability to trigger a Th1-predominant response in both murine and human subjects. Our past studies have shown that the injection of a rEA-encoding adenovirus (rAd5-rEA) alongside an HIV antigen-encoding adenovirus greatly improves the adaptive immune response against this pathogen-derived transgene. In this report, we investigated whether rAd5-rEA could promote and/or alter cytotoxic memory responses toward carcinoembryonic antigen (CEA), a colorectal cancer-related TAA. We found that the addition of rAd5-rEA to an Ad-based CEA vaccine induced a dose-dependent increase in several anti-CEA T and B cell responses. Moreover, inclusion of rAd5-rEA increased the number of CEA-derived antigenic epitopes that elicited significant cell-mediated and IgG-mediated recognition. These enhanced anti-CEA immune responses also translated into superior CEA-targeted cell killing, as evaluated by an in vivo cytotoxic T lymphocyte assay. Overall, these results suggest that co-administration of rAd5-rEA with a tumor antigen vaccine can substantially boost and broaden the TAA-specific adaptive memory response, thereby validating the potential of rAd5-rEA to be a beneficial adjuvant during therapeutic cancer vaccination.

  10. Construction and Application of an Avian Intestinal Intraepithelial Lymphocyte cDNA Microarray for Gene Expression Profiling during Eimeria maxima Infection

    USDA-ARS?s Scientific Manuscript database

    Intestinal intraepithelial lymphocytes (IELs) are the primary immune effector cells in the gut and play a critical role in eliciting protective immunity to enteric pathogens such as Eimeria, the etiologic agent of avian coccidiosis. In this study, a library of genes expressed by intestinal IELs of ...

  11. Functional characterization of tumor necrosis factor superfamily 15 (TNFSF15) induced by lipopolysaccharides and Eimeria infection.

    PubMed

    Park, Soon S; Lillehoj, Hyun S; Hong, Yeong Ho; Lee, Sung Hyen

    2007-01-01

    A full-length cDNA encoding chicken tumor necrosis factor superfamily 15 (TNFSF15) was isolated and its functional role was investigated. TNFSF15 transcripts were primarily expressed in spleen, liver, intestinal intraepithelial lymphocytes (IEL), peripheral blood lymphocytes and bursa. In vitro infection of HTC macrophages with three species of Eimeria sporozoites induced TNFSF15 gene expression. In vivo experiments revealed that TNFSF15 gene was highly increased following primary infections with Eimeria acervulina or Eimeria maxima. In contrast, no consistent changes in transcript levels were seen following primary infection with Eimeria tenella, or following secondary infection with any of the three Eimeria species. Following infection with E. acervulina and E. maxima, TNFSF15 transcripts were primarily expressed in intestinal CD4(+) and TCR2(+) IEL, respectively. A dose-dependent cytotoxic effect of recombinant TNFSF15 protein was observed on HTC and LSCC-RP9 tumor cells. These results indicate that TNFSF15 plays an important role in local inflammatory response to Eimeria.

  12. Construction and immunological evaluation of recombinant Lactobacillus plantarum expressing SO7 of Eimeria tenella fusion DC-targeting peptide.

    PubMed

    Yang, Guilian; Yao, Jiayun; Yang, Wentao; Jiang, Yanlong; Du, Jinfen; Huang, Haibin; Gu, Wei; Hu, Jingtao; Ye, Liping; Shi, Chunwei; Shan, Baolong; Wang, Chunfeng

    2017-03-15

    The coccidiosis caused by Eimeria tenella (coccidian) and other species is a serious parasitic disease that affects the global poultry breeding industry. Lactobacillus strains exhibit a number of properties that make them attractive candidates as delivery vehicles for presentation to the mucosa of compounds with pharmaceutical interest, particularly vaccines. Here, the recombinant Lactobacillus plantarum (co-expressing SO7 and DCpep gene) was constructed, and its efficacy against E. tenella challenge was evaluated in this study. Broiler chickens were orally immunized with live recombinant L. plantarum NC8-pSIP409-SO7-DCpep for two weeks and were then challenged with 5×10(4)E.tenella sporulated oocysts per chicken. During the experiment, body weight gains, cecum lesion scores, fecal oocyst shedding and antibody responses in serum and intestinal washes were assessed as measures of protective immunity. The results indicated that chickens immunized with live recombinant L. plantarum can increase body weight gains and serum antibody responses compared to the control groups. Meanwhile, fecal oocyst shedding in the immunized group was significantly reduced (p<0.01). Moreover, recombinant L. plantarum can significantly relieve pathological damage in cecum, according to lesion scores and histopathologic cecum sections (p<0.01). Therefore, these results indicate that recombinant L. plantarum NC8-pSIP409-SO7-DCpep could become a promising oral vaccine candidate against E. tenella infection.

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

    PubMed Central

    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. PMID:26641262

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

    PubMed

    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.

  15. Immunogenicity of recombinant attenuated Salmonella enterica serovar Typhimurium vaccine strains carrying a gene that encodes Eimeria tenella antigen SO7.

    PubMed

    Konjufca, Vjollca; Jenkins, Mark; Wang, Shifeng; Juarez-Rodriguez, Maria Dolores; Curtiss, Roy

    2008-12-01

    Recombinant attenuated Salmonella vaccines against avian coccidiosis were developed to deliver Eimeria species antigens to the lymphoid tissues of chickens via the type 3 secretion system (T3SS) and the type 2 secretion system (T2SS) of Salmonella. For antigen delivery via the T3SS, the Eimeria tenella gene encoding sporozoite antigen SO7 was cloned downstream of the translocation domain of the Salmonella enterica serovar Typhimurium sopE gene in the parental pYA3868 and pYA3870 vectors to generate pYA4156 and pYA4157. Newly constructed T3SS vectors were introduced into host strain chi8879 (Delta phoP233 Delta sptP1033::xylE Delta asdA16), an attenuated derivative of the highly virulent UK-1 strain. The SopE-SO7 fusion protein was secreted by the T3SS of Salmonella. The vector pYA4184 was constructed for delivery of the SO7 antigen via the T2SS. The SO7 protein was toxic to Salmonella when larger amounts were synthesized; thus, the synthesis of this protein was placed under the control of the lacI repressor gene, whose expression in turn was dependent on the amount of available arabinose in the medium. The pYA4184 vector was introduced into host strain chi9242 (Delta phoP233 Delta asdA16 Delta araBAD23 Delta relA198::araC P(BAD) lacI TT [TT is the T4ipIII transcription terminator]). In addition to SO7, for immunization and challenge studies we used the EAMZ250 antigen of Eimeria acervulina, which was previously shown to confer partial protection against E. acervulina challenge when it was delivered via the T3SS. Immunization of chickens with a combination of the SO7 and EAMZ250 antigens delivered via the T3SS induced superior protection against challenge by E. acervulina. In contrast, chickens immunized with SO7 that was delivered via the T2SS of Salmonella were better protected from challenge by E. tenella.

  16. A novel recombinant BCG vaccine encoding eimeria tenella rhomboid and chicken IL-2 induces protective immunity against coccidiosis.

    PubMed

    Wang, Qiuyue; Chen, Lifeng; Li, Jianhua; Zheng, Jun; Cai, Ning; Gong, Pengtao; Li, Shuhong; Li, He; Zhang, Xichen

    2014-06-01

    A novel recombinant Bacille Calmette-Guerin (rBCG) vaccine co-expressed Eimeria tenella rhomboid and cytokine chicken IL-2 (chIL-2) was constructed, and its efficacy against E. tenella challenge was observed. The rhomboid gene of E. tenella and chIL-2 gene were subcloned into integrative expression vector pMV361, producing vaccines rBCG pMV361-rho and pMV361-rho-IL2. Animal experiment via intranasal and subcutaneous route in chickens was carried out to evaluate the immune efficacy of the vaccines. The results indicated that these rBCG vaccines could obviously alleviate cacal lesions and oocyst output. Intranasal immunization with pMV361-rho and pMV361-rho-IL2 elicited better protective immunity against E. tenella than subcutaneous immunization. Splenocytes from chickens immunized with either rBCG pMV361-rho and pMV361-rho-IL2 had increased CD4(+) and CD8(+) cell production. Our data indicate recombinant BCG is able to impart partial protection against E. tenella challenge and co-expression of cytokine with antigen was an effective strategy to improve vaccine immunity.

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

    PubMed

    Almeida, Gustavo F D; Thamsborg, Stig M; Madeira, Alda M B N; Ferreira, Jorge F S; Magalhães, Pedro M; Demattê Filho, Luiz C; Horsted, Klaus; Hermansen, John E

    2014-03-01

    Due to an increasing demand for natural products to control coccidiosis in broilers, we investigated the effects of supplementing a combination of ethanolic extracts of Artemisia annua and Curcuma longa in drinking water. Three different dosages of this herbal mixture were compared with a negative control (uninfected), a positive control (infected and untreated), chemical coccidiostats (nicarbazin+narazin and, later, salinomycin), vaccination, and a product based on oregano. Differences in performance (weight gain, feed intake, and feed conversion rate), mortality, gross intestinal lesions 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 in broilers given vaccination, oregano and in negative controls. There was a trend for lower mortality (P = 0·08) in the later stage of the growing period (23-43 days) in broilers given the highest dosage of herbal mixture compared with broilers given chemical coccidiostats. In conclusion, the delivery strategy of the herbal extracts is easy to implement at farm level, but further studies on dose levels and modes of action are needed.

  18. The immunodominant Eimeria acervulina sporozoite antigen previously described as p160/p240 is a 19-kilodalton antigen present in several Eimeria species.

    PubMed

    Laurent, F; Bourdieu, C; Kazanji, M; Yvoré, P; Péry, P

    1994-01-01

    A lambda Zap II cDNA expression library, constructed from Eimeria acervulina (PAPa46 strain) sporulated oocyst stage, was screened with sera raised to E. acervulina or Eimeria tenella oocysts in order to isolate clones coding for antigens common to the two species. Most of the clones isolated were derived from the same gene. Antisera raised to a recombinant glutathione-S-transferase fusion protein 1P reacted with an antigen of 19 kDa in immunoblot of E. acervulina sporulated and unsporulated oocysts. Immunofluorescence of E. acervulina sporozoites indicated that the antigen is located in the cytoplasm. The anti-1P antisera reacted on immunoblots of E. tenella with a 19-kDa antigen and by immunofluorescence on E. tenella, Eimeria maxima and Eimeria falciformis sporozoites, indicating that the antigen is conserved in Eimeria species. DNA sequencing indicated that the sequence was almost identical to that of clone cSZ1 previously described by Jenkins et al. using E. acervulina strain #12. The 1P insert hybridized to a 1150-nt mRNA from E. acervulina PAPa46 strain and strain #12, a size consistent with the observed molecular weight of the protein.

  19. In ovo vaccination with the Eimeria tenella EtMIC2 gene induces protective immunity against coccidiosis.

    PubMed

    Ding, Xicheng; Lillehoj, Hyun S; Dalloul, Rami A; Min, Wongi; Sato, Takanori; Yasuda, Atsushi; Lillehoj, Erik P

    2005-05-25

    An Eimeria tenella microneme recombinant gene (EtMIC2) and encoded protein were evaluated as potential vaccines against avian coccidiosis. In ovo inoculation with the EtMIC2 gene increased anti-EtMIC2 antibody titers at days 10 and 17 following E. tenella infection. In addition, vaccinated birds developed protective immunity against infection by E. tenella as assessed by significantly increased body weight gain and decreased fecal oocyst shedding compared with non-vaccinated controls. Vaccination with the EtMIC2 gene also led to protective immunity against infection by E. acervulina, but not E. maxima. Combined in ovo DNA vaccination plus post-hatch boosting with EtMIC2 DNA or protein did not improve antibody titers or protective immunity beyond that achieved with in ovo vaccination alone. These results provide evidence that in ovo immunization with a recombinant Eimeria microneme gene stimulates protective intestinal immunity against coccidiosis.

  20. Molecular characterization and phylogenetic analysis of Eimeria from turkeys and gamebirds: implications for evolutionary relationships in Galliform birds.

    PubMed

    Miska, K B; Schwarz, R S; Jenkins, M C; Rathinam, T; Chapman, H D

    2010-10-01

    In order to determine the evolutionary relationships among Eimeria species that parasitize birds of the Galliformes, the 18s rDNA gene and a portion of the cytochrome oxidase subunit 1 (cox-1) were amplified from Eimeria species isolated from turkeys, chukars, and pheasants. The phylogenetic analysis of these sequences suggests that species infecting chickens are polyphyletic and, therefore, do not all share a direct common ancestor. Both the 18s rDNA and the cox-1 sequences indicate that Eimeria tenella and Eimeria necatrix are more closely related to Eimeria of turkeys and pheasants than to other species that infect the chicken. It is, therefore, likely that the chicken Eimeria spp. represent 2 separate ancestral colonizations of the gut, one of which comprises E. tenella and E. necatrix that infect the ceca, while the other includes Eimeria acervulina, Eimeria brunetti, Eimeria maxima, and Eimeria mitis, which infect the upper regions of the intestine.

  1. 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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    PubMed Central

    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

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

    PubMed

    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/

  4. Evaluation of immune protective efficacies of Eimeria tenella EtMic1 polypeptides with different domain recombination displayed on yeast surface.

    PubMed

    Chen, Peipei; Lv, Junfeng; Zhang, Jie; Sun, Hui; Chen, Zhengtao; Li, Hongmei; Wang, Fangkun; Zhao, Xiaomin

    2015-08-01

    In the present study, three different live oral vaccines using the EBY100/pCTCON-2 yeast surface display system with different Eimeria tenella microneme-1 (EtMic1) protein domain recombination were constructed and their protective efficacies against homologous challenge were compared by evaluating the body weight gains, relative growth rate, cecal lesion scores, oocyst output, oocyst decrease ratio, anti-coccidial index, the serum antibody levels and the proliferation ability of blood lymphocytes. The results indicated that all the three constructed live oral vaccines expressing different EtMic1 polypeptides provided excellent protection against homologous challenge by significantly increasing weight gains, reducing cecal lesions, achieving a high ACI, elevating specific antibody response and splenocyte proliferation ability compared with controls. The yeasts displaying the EtMic1 polypeptide-III (expressed TSP-2, TSP-3 and TSP-4 domains) provided better protection against challenge than the yeasts displaying either the EtMic1 polypeptide-I (expressed I-domain, TSP-1 and TSP-2) or polypeptide-II (expressed I-domain and all the five TSP domains) did. Considering the exclusion of antibiotic resistant gene in the system, the strain EBY100 of Saccharomyces cerevisiae may be a better choice for coccidian antigen delivery.

  5. Eimeria in Fowls

    PubMed Central

    Rose, M. Elaine; Long, P. L.

    1962-01-01

    Two or three graded infections with oocysts of Eimeria acervulina, E. tenella, E. necatrix and E. maxima produced a resistance to further infection with the immunizing species. The oocyst output after the second infection, in each case, was lower than that after the initial dose indicating the substantial immunizing effect of the initial infection. The species could be placed in a descending order of immunizing activity as follows: E. maxima, E. acervulina, E. tenella and E. necatrix. A solid immunity to the immunizing species in no way prevented the development of an additional infection, here referred to as `cross-infection', with any of the species studied. Serum precipitins were produced in infections with all four species, the response to infection with E. necatrix being less marked than to the other species. A first challenge of immune fowls with the immunizing species produced some increase in precipitation in agar whereas a second challenge had no such effect; the significance of this lack of response is discussed. Usually, fowls immunized against one species and then infected with an additional one, produced serum precipitins which reacted only with the antigen of the additional species. But E. tenella immunized fowls, when given an additional infection with E. necatrix, produced precipitins that reacted with antigens of both species. The same was also true when E. necatrix immunized fowls were infected with E. tenella. ImagesFIG. 9 PMID:14493839

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

    PubMed Central

    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

  7. Application of polymerase chain reaction based on ITS1 rDNA to speciate Eimeria.

    PubMed

    Jenkins, M C; Miska, K; Klopp, S

    2006-03-01

    A method was developed to recover Eimeria spp. oocysts directly from poultry litter and determine which species of Eimeria were present using polymerase chain reaction (PCR) based on the ITS1 rDNA sequence. The species composition of Eimeria oocysts was also compared before and after propagation in susceptible chickens to determine if the relative proportion of each species changed after expansion. In samples from two broiler operations, ITS1-PCR was able to detect Eimeria spp. oocysts recovered from litter, with Eimeria acervulina, Eimeria maxima, and Eimeria praecox being the predominant species present therein. Although Eimeria tenella was found in one sample, the other species--Eimeria brunetti, Eimeria necatrix, and Eimeria mitis-were not detected. The species composition as determined by ITS1-PCR did not appear to appreciably alter after expansion in susceptible chickens. The described method represents a rapid means for determining the major Eimeria species in a poultry operation and may be helpful in choosing a particular live oocyst vaccine formulation to protect chickens against coccidiosis.

  8. Characterization of monoclonal antibodies that recognize the Eimeria tenella microneme protein MIC2.

    PubMed

    Sasai, Kazumi; Fetterer, Raymond H; Lillehoj, Hyun; Matusra, Satomi; Constantinoiu, Constantin C; Matsubayashi, Makoto; Tani, Hiroyuki; Baba, Eiichiroh

    2008-12-01

    The apicomplexan pathogens of Eimeria cause coccidiosis, an intestinal disease of chickens, which has a major economic impact on the poultry industry. Members of the Apicomplexa share an assortment of unique secretory organelles (rhoptries, micronemes and dense granules) that mediate invasion of host cells and formation and modification of the parasitophorous vacuole. Among these, microneme protein 2 from Eimeria tenella(EtMIC2) has a putative function in parasite adhesion to the host cell to initiate the invasion process. To investigate the role of EtMIC2 in host parasite interactions, the production and characterization of 12 monoclonal antibodies (mabs) produced against recombinant EtMIC2 proteins is described. All mabs reacted with molecules belonging to the apical complex of sporozoites and merozoites of E. tenella, E. acervulina and E. maxima in an immunofluorescence assay. By Western blot analysis, the mabs identified a developmentally regulated protein of 42 kDa corresponding to EtMIC 2 and cross-reacted with proteins in developmental stages of E. acervulina. Collectively, these mabs are useful tools for the detailed investigation of the characterization of EtMIC2 related proteins in Eimeria species.

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

    PubMed

    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.

  10. Improved polymerase chain reaction technique for determining the species composition of Eimeria in poultry litter.

    PubMed

    Jenkins, M C; Miska, K; Klopp, S

    2006-12-01

    An improved polymerase chain reaction (PCR)-based method for determining the species composition of Eimeria in poultry litter was developed by incorporating species-specific internal standards in the assay. Internal standard molecules were prepared by fusing seven different Eimeria species-specific intervening transcribed sequence 1 (ITS1) rDNA primer pairs to a non-Eimeria DNA molecule and by cloning the hybrid DNA molecules into a plasmid. The internal DNA standards were then used in Eimeria-specific ITS 1 PCR, and they were found to be capable of detecting E. acervulina, E. maxima, E. praecox, and E. tenella oocysts isolated directly from poultry litter.

  11. Eimeria stiedae

    PubMed Central

    Rose, M. Elaine

    1959-01-01

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

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

    PubMed

    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.

  13. Identification and characterization of an Eimeria-conserved protein in Eimeria tenella.

    PubMed

    Dong, Hui; Wang, Yange; Han, Hongyu; Li, Ting; Zhao, Qiping; Zhu, Shunhai; Li, Liujia; Wu, Youling; Huang, Bing

    2014-02-01

    The precocious lines of Eimeria spp. have unique phenotypes. However, the genetic basis of the precocious phenotype is still poorly understood. The identification of Eimeria genes controlling the precocious phenotype is of immense importance in the fight against coccidiosis. In the present study, a novel gene of Eimeria maxima was cloned using rapid amplification of cDNA ends (RACE) based on the expressed sequence tag (EST). Homologous genes were also found in Eimeria tenella and Eimeria acervulina. Alignment of the amino acid sequences from E. tenella, E. maxima, and E. acervulina showed 80-86 % identity, demonstrating a conserved protein in different Eimeria spp. This gene, designated Eimeria-conserved protein (ECP), contained 235 amino acids with a predicted molecular mass of 25.4 kDa and had 100 % identity with one annotated protein from E. maxima (Emax_0517). Real-time PCR and Western blot analysis revealed that the expression of ECP at mRNA and protein level in E. tenella is developmentally regulated. Messenger RNA levels from the ECP gene were higher in sporozoites than in other developmental stages (unsporulated oocysts, sporulated oocysts, and second-generation merozoites). Expression of ECP protein was detected in unsporulated oocysts, increased in abundance in sporulated oocysts, and was most prominent in sporozoites. Thereafter, the level of the ECP protein decreased, and no ECP-specific protein was detected in second-generation merozoites. Immunostaining with anti-rECP indicated that ECP is highly concentrated in both refractile bodies (RB) of free sporozoites, but is located at the apical end of the sporozoites after invasion of DF-1 cells. The specific staining of the ECP protein becomes more intense in trophozoites and immature first-generation schizonts, but decreases in mature first-generation schizonts. Inhibition of the function of ECP using specific antibodies reduced the ability of E. tenella sporozoites to invade host cells. Compared with the

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

    PubMed

    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

  15. Protective immunity against Eimeria tenella infection in chickens induced by immunization with a recombinant C-terminal derivative of EtIMP1.

    PubMed

    Yin, Guangwen; Lin, Qian; Wei, Wenjun; Qin, Mei; Liu, Xianyong; Suo, Xun; Huang, Zhijian

    2014-12-15

    Immune mapped protein-1 (IMP1) is a new protective protein in apicomplexan parasites, and exits in Eimeria tenella. Cloning and sequence analysis has predicted the antigen to be a novel membrane protein of apicomplexan parasites. In order to assess the immunogenicity of EtIMP1, a C-terminal derivative of EtIMP1 was expressed in a bacterial host system and was used to immunize chickens. The protective efficacy against a homologous challenge was evaluated by body weight gains, lesion scores and fecal oocyst shedding. The results showed that the subunit vaccine can improve weight gains, reduced cecal pathology and lower oocyst fecal shedding compared with non immunized controls. The results suggested that the C-terminal derivative of EtIMP1 might be considered as a candidate in the development of subunit vaccines against Eimeria infection.

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

    PubMed

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

    2012-06-01

    Vaccines composed of either virulent or attenuated Eimeria spp. oocysts have been developed as an alternative to medication of feed with ionophore drugs or synthetic chemicals. The purpose of this study was to evaluate the use of gel-beads containing a mixture of Eimeria acervulina, Eimeria maxima, and Eimeria tenella oocysts as a vaccine against coccidiosis. Newly hatched chicks (Gallus gallus domesticus) were either sprayed with an aqueous suspension of Eimeria oocysts or were allowed to ingest feed containing Eimeria oocysts-incorporated gel-beads. Control day-old chicks were given an equivalent number of Eimeria oocysts (10(4) total) by oral gavage. After 3 days, chicks were randomly assigned to individual cages, and feces were collected between days 5 and 8 postinfection. All samples were processed for total Eimeria oocysts. At 4 wk of age, all chickens and a control nonimmunized group received a high-dose E acervulina, E maxima, and E. tenella challenge infection. Oocyst excretion by chicks fed gel-beads or inoculated by oral gavage was 10- to 100-fold greater than that of chicks spray-vaccinated with the Eimeria oocysts mixture (log 6.3-6.6 vs. log 4.8). Subsequent protection against challenge as measured by weight gain and feed conversion efficiency was significantly greater (P < 0.05) in gel-bead and oral gavage groups compared with spray-vaccinated or nonimmunized groups. Also, gel-bead and oral gavage groups showed no significant difference (P > 0.05) in weight gain and feed conversion efficiency compared with nonchallenged controls. These findings indicate that incorporation of Eimeria spp. oocysts in gel-beads may represent an effective way to deliver live oocyst vaccines to day-old chicks for preventing subsequent outbreaks of coccidiosis in the field.

  17. The molecular characterization and immune protection of microneme 2 of Eimeria acervulina.

    PubMed

    Zhang, ZhenChao; Liu, LianRui; Huang, JingWei; Wang, Shuai; Lu, MingMin; Song, XiaoKai; Xu, LiXin; Yan, RuoFeng; Li, XiangRui

    2016-01-15

    Eimeria acervulina (E. acervulina) is an intestinal protozoon and few of its antigen genes were researched. In this study, the gene of E. acervulina microneme protein 2 (EaMIC2) was cloned and characterized. The degenerate primers for the conserved sequence of microneme protein 2 were designed based on Eimeria tenella microneme 2 (EtMIC2) (AF111839.1), Eimeria maxima microneme 2 (EmMIC2) (FR718971.1) and E. brunetti microneme 2 (EbMIC2) (AB723700.1) to amplify the conserved sequence of microneme protein 2. According to the conserved sequence, specific primers for the rapid amplification of cDNA ends (RACE) were designed to amplify the 3'- and 5'-ends of EaMIC2. The full length cDNA of this gene was obtained by overlapping the sequences of 3'- and 5'-extremities and amplification by reverse transcription PCR. The sequence analysis revealed that the opening reading frame (ORF) of EaMIC2 was 882bp and encoded a protein of 293 amino acids with 29.81kDa. The most part of the mature protein (base: 82-882bp, amino acid: 28-293aa) of EaMIC2 (mpmEaMIC2) was inserted into pCold TF to produce recombinant mpmEaMIC2. Western blotting assay was used to analysis the immunogenicity of mpmEaMIC2 and the result showed that the recombinant protein was successfully recognized by the sera of chickens experimentally infected with E. acervulina, while the native protein in the somatic extract of sporozoites was as well detected by sera from rats immunized with the recombinant protein of mpmEaMIC2. Immunofluorescence analysis using antibody against recombinant protein mpmEaMIC2 indicated that this protein was expressed in the sporozoites and merozoites stages of E. acervulina. Animal challenge experiments demonstrated that the recombinant protein of mpmEaMIC2 could significantly increase the average body weight gains, decrease the mean lesion scores and the oocyst outputs of the immunized chickens. All the above results suggested that the EaMIC2 was a novel E. acervulina antigen and could

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

    PubMed Central

    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

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

    PubMed

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

    PubMed

    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.

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

    USDA-ARS?s Scientific Manuscript database

    Two phytonutrient mixtures, VAC (carvacrol, cinnamaldehyde, and Capsicum oleoresin), and MC (Capsicum oleoresin and turmeric oleoresin), were evaluated for their effects on local and systemic immune responses following immunization of chickens with an Eimeria recombinant protein. Chickens were fed ...

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

    PubMed Central

    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

  3. Cloning and characterization of an Eimeria acervulina sporozoite gene homologous to aspartyl proteinases.

    PubMed

    Laurent, F; Bourdieu, C; Kaga, M; Chilmonczyk, S; Zgrzebski, G; Yvoré, P; Péry, P

    1993-12-01

    A lambda ZapII cDNA library was constructed using mRNA from Eimeria acervulina sporulated oocysts and screened with monoclonal antibodies raised against Eimeria tenella sporulated oocytes. Monoclonal antibody N3C8B12 identified a clone (6S2) potentially encoding an aspartyl proteinase since significant homology with cathepsin D, pepsin and renin proteinases was revealed by sequence comparisons. The 1500-bp cDNA fragment containing the coccidial gene was subcloned into pGEX-FA expression vector, leading to the production of an 80-kDa fusion protein (FA6S2) which was used to immunize rabbits. The anti-FA6S2 rabbit sera revealed a single 43-kDa protein present in Eimeria acervulina, Eimeria tenella, Eimeria maxima and Eimeria falciformis sporulated oocyst antigens. Indirect immunofluorescence and electron microscopy with mAb N3C8B12 localized the putative aspartyl proteinase in the refractile bodies of Eimeria tenella sporozoites.

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  6. Gel-Bead Delivery of Eimeria Oocysts Protects Chickens Against Coccidiosis

    USDA-ARS?s Scientific Manuscript database

    Vaccines composed of either virulent or attenuated Eimeria spp. oocysts have been developed as an alternative to medication of feed with ionophore drugs or synthetic chemicals. The purpose of this study was to evaluate the use of gel beads containing a mixture of E. acervulina, E. maxima, and E. te...

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

    USDA-ARS?s Scientific Manuscript database

    Control of avian coccidiosis is increasingly being achieved by the administration of low doses of Eimeria oocysts to newly hatched chicks. The purpose of this study was to test the efficacy of gel-beads containing a mixture of E. acervulina, E. maxima, and E. tenella oocysts as a vaccine to protect ...

  8. Eimeria tenella heat shock protein 70 enhances protection of recombinant microneme protein MIC2 subunit antigen vaccination against E. tenella challenge.

    PubMed

    Zhang, Lei; Ma, Liping; Liu, Renqiang; Zhang, Yunfei; Zhang, Shouping; Hu, Chunmei; Song, Meng; Cai, Jianping; Wang, Ming

    2012-09-10

    Heat shock proteins have been reported to stimulate the immune system via innate receptors. Our study found that the novel immunopotentiator, Eimeria tenella (E. tenella) heat shock protein 70 (HSP70), enhanced protective immunity elicited by E. tenella antigen microneme protein 2 (EtMIC2) against avian coccidiosis. It demonstrated that the expression of TLR2 and TLR4 were strongly upregulated in EtHSP70 and EtMIC2 plus EtHSP70 stimulated chicken embryo fibroblasts (CEF) compared with untreated controls and EtMIC2 alone. In addition, the same treatment induced high levels of interleukin (IL)-12 and interferon (IFN)-γ that are critical cytokines of innate immunity. In vivo experiments involved using broiler chickens subcutaneously immunized with EtMIC2 alone or EtMIC2 plus EtHSP70 at 7 and 14 days post-hatch, which were then orally challenged with live E. tenella at 7 days following secondary immunization. Body weight gains, cecal lesion scores, fecal oocyst shedding, serum antibody responses against MIC2, and intestinal cytokine transcript levels were assessed as measures of protective immunity. Chickens immunized with EtMIC2 plus EtHSP70 showed increased body weight gains, decreased oocyst shedding, increased serum antibody responses, and high levels of IL-12, IFN-γ, and IL-17 compared with the EtMIC2 only or control groups. Moreover, chickens immunized with EtHSP70 alone showed significantly protective effect against E. tenella infection. In summary, this study provides the first evidence of the immunoenhancing activities of EtHSP70 in poultry.

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

    PubMed

    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) greater weight gain (WG) compared to nonimmunized controls. Feed conversion ratio (FCR) also showed a significant (P < 0.05) improvement in both groups relative to nonimmunized controls. Moreover, WG and FCR in both groups was not significantly different (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.

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

    PubMed

    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% (P<0.05) in relation to the control group. Challenged groups presented a decrease (P<0.05) in weight gain compared with control groups. All challenged groups showed a reduction in weight gain, even when there was no reduction (P<0.05) in feed intake (adjustment through variance-covariance analysis). The feed intake variation in broilers infected with E. acervulina, E. maxima, E. tenella, or Pool showed a quadratic (P<0.05) influence over the variation in weight gain. In relation to the isolated effects, the challenges have an impact of less than 1% over the variance in feed intake and weight gain. However, the magnitude of the effects varied with Eimeria 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.

  11. Effects of novel adjuvant complex/Eimeria profilin vaccine on intestinal host immune responses against live E. acervulina challenge infection

    USDA-ARS?s Scientific Manuscript database

    The effects of a novel adjuvant; composed of Quil A, cholesterol, dimethyl dioctadecyl ammonium bromide, and Carbopol (QCDC), on protective immunity against avian coccidiosis following immunization with an Eimeria recombinant protein were determined. Broiler chickens were subcutaneously immunized w...

  12. A simplified protocol for molecular identification of Eimeria species in field samples.

    PubMed

    Haug, Anita; Thebo, Per; Mattsson, Jens G

    2007-05-15

    This study aimed to find a fast, sensitive and efficient protocol for molecular identification of chicken Eimeria spp. in field samples. Various methods for each of the three steps of the protocol were evaluated: oocyst wall rupturing methods, DNA extraction methods, and identification of species-specific DNA sequences by PCR. We then compared and evaluated five complete protocols. Three series of oocyst suspensions of known number of oocysts from Eimeria mitis, Eimeria praecox, Eimeria maxima and Eimeria tenella were prepared and ground using glass beads or mini-pestle. DNA was extracted from ruptured oocysts using commercial systems (GeneReleaser, Qiagen Stoolkit and Prepman) or phenol-chloroform DNA extraction, followed by identification of species-specific ITS-1 sequences by optimised single species PCR assays. The Stoolkit and Prepman protocols showed insufficient repeatability, and the former was also expensive and relatively time-consuming. In contrast, both the GeneReleaser protocol and phenol-chloroform protocols were robust and sensitive, detecting less than 0.4 oocysts of each species per PCR. Finally, we evaluated our new protocol on 68 coccidia positive field samples. Our data suggests that rupturing the oocysts by mini-pestle grinding, preparing the DNA with GeneReleaser, followed by optimised single species PCR assays, makes a robust and sensitive procedure for identifying chicken Eimeria species in field samples. Importantly, it also provides minimal hands-on-time in the pre-PCR process, lower contamination risk and no handling of toxic chemicals.

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

    PubMed

    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.

  14. Effects of simple and disposable chicken cages for experimental Eimeria infections.

    PubMed

    Yoo, Jeongmi; Kang, Sung H; Jeong, Jipseol; Kim, Woo H; Kim, Suk; Lillehoj, Hyun S; Min, Wongi

    2011-09-01

    During experimental Eimeria infections in chickens, facilities are often contaminated by fecal oocysts known to be highly resistant to both chemical and enzymatic treatments. Thus, studies using experimental Eimeria infections have been limited due to the difficulty of complete elimination of residual oocysts from both cages and facilities. To overcome this limitation, simple, inexpensive, and disposable cages were constructed from cardboard boxes and tested during experimental Eimeria maxima infections. The cages were used in animal rooms with only a 1.7% evidence of coccidia contamination between adjacent cages. No significant differences in fecal oocyst output and body weight gain were noted between animals housed in disposable cages and animals housed in wire control cages. This cage design is a useful means for preventing oocyst contamination during experimental conditions, suggesting that this disposable cage design could be used for other avian infectious disease studies.

  15. Observations on the gross pathology of Eimeria praecox infections in chickens.

    PubMed

    Allen, P C; Jenkins, M C

    2010-06-01

    Challenge infections with 10(3), 5 x 10(4), 10(5), or 5 x 10(5) sporulated Eimeria praecox oocysts caused moderate but significant weight gain reduction at all infective doses. Substantial reduction in plasma carotenoids and moderate but significant increases in plasma NO2(-) + NO3- were observed only at the two higher doses when measured at day 6 postchallenge (PC). Daily monitoring of chickens after challenge with 5 x 10(4) oocysts revealed an inflammatory response in the duodenum and jejunum beginning at day 1 PC that was associated with a significant increase in levels of plasma NO2(-) + NO3-, which peaked at day 4 PC. A moderate, uniform hyperplasia of the small intestine and significant depression of plasma carotenoids were observed on days 4-6 PC. Plasma NO2(-) + NO3- decreased to control levels by day 6 PC. All infections were accompanied by production of a mucoid exudate in the duodenum and jejunum, which became thick and opaque by 4 days PC and tended to obscure mildly inflamed areas. These observations indicate that the acute host response to primary infection with E. praecox is both different from and occurs earlier than the response to experimental infections with other Eimeria spp., such as Eimeria acervulina, Eimeria maxima, or Eimeria tenella. These factors need to be considered in observations of pathology arising from co-infections of E. praecox with other Eimeria species, especially in drug sensitivity testing of Eimeria oocysts recovered from litter and in the evaluation of live oocyst vaccines.

  16. Metam sodium reduces viability and infectivity of Eimeria oocysts.

    PubMed

    Fetterer, R H; Jenkins, M C; Miska, K B; Cain, G D

    2010-06-01

    Metam sodium (MS, sodium N-methyldithiocarbamate) is a widely used soil pesticide. Fumigation or chemical sterilization of poultry litter containing infectious oocysts could be an effective strategy to block the transmission of avian coccidia. In the current study, the effect of MS on the viability and infectivity of ocysts was investigated. The development of isolated, unsporulated oocysts of both Eimeria tenella and Eimeria maxima was inhibited, in a dose-related manner (IC(50) 8 to 14 microg/ml), by exposure to aqueous MS. Most treated oocysts failed to develop beyond early stages of sporulation. To determine the effect of MS on infectivity, isolated oocysts of E. tenella , Eimeria acervulina , and E. maxima were exposed for 24 hr to aqueous concentrations of MS ranging from 0 to 1,000 microg/ml. Treated oocysts were inoculated into chickens, and parameters of coccidiosis infection were compared to chickens inoculated with equal numbers of untreated oocysts. In a dose-related manner, MS significantly reduced the infectivity of oocysts with maximum effect observed at a dose of 300 microg/ml. When a mixture of oocysts containing 3 coccidian species was exposed to 300 microg/ml MS, from 0 to 24 hr, infectivity of oocysts was significantly reduced after a minimum of 12 hr of exposure. Treatment of aqueous slurries of litter samples obtained from commercial poultry houses, with 300 microg/ml MS for 24 hr, prevented the sporulation of eimerian oocysts in the litter samples relative to untreated control samples. The results indicate that MS could be used to reduce coccidial contamination of poultry litter.

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

    PubMed

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

    2014-05-28

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

  18. Montanide ISA 71 VG adjuvant enhances antibody and cell-ediated immune responses to profilin subunit antigen vaccination and promotes protection against Eimeria acervulina and Eimeria tenella

    USDA-ARS?s Scientific Manuscript database

    The present study was conducted to investigate the immunoenhancing effects of ISA 71 VG adjuvant on profilin subunit antigen vaccination. Broiler chickens were immunized subcutaneously with a purified Eimeria acervulina recombinant profilin protein, either alone or mixed with ISA 71 VG, and host imm...

  19. Genetic diversity within ITS-1 region of Eimeria species infecting chickens of north India.

    PubMed

    Kumar, Saroj; Garg, Rajat; Banerjee, P S; Ram, Hira; Kundu, K; Kumar, Sunil; Mandal, M

    2015-12-01

    Coccidiosis, caused by apicomplexan parasites of the genus Eimeria, inflicts severe economic losses to the poultry industry around the globe. In the present study, ITS-1 based species specific nested PCR revealed prevalence of E. acervulina, E. brunetti, E. maxima, E. mitis, E. praecox, E. necatrix and E. tenella in 79.2%, 12.5%, 64.6%, 89.6%, 60.4%, 64.6% and 97.9% poultry farms of north India, respectively. The ITS-1 sequences of different Eimeria spp. from north India were generated and analyzed to establish their phylogenetic relationship. The sequence identity with available sequences ranged from 80 to 100% in E. tenella, 95 to 100% in E. acervulina, 64 to 97% in E. necatrix, 96 to 99% in E. brunetti and 97 to 98% in E. mitis. Only long ITS-1 sequences of E. maxima could be generated in the present study and it had 80-100% identity with published sequences. Two out of the four ITS-1 sequences of E. maxima had mismatches in the published nested primer sequences from Australia, while one sequence of E. necatrix had a mismatch near 3' end of both forward and reverse published nested primer sequences, warranting for the need of designing new set of degenerate primers for these two species of Eimeria. In the phylogenetic tree, all isolates of E. acervulina, E. brunetti, E. mitis, E. tenella and E. necatrix clustered in separate clades with high bootstrap value. E. maxima sequences of north Indian isolates grouped in a long form of E. maxima clade. Complete ITS-1 sequences of E. necatrix and E. mitis are reported for the first time from India. Further studies are required with more number of isolates to verify whether these differences are unique to geographical locations.

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

    PubMed

    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

  1. In ovo vaccination using Eimeria profilin and Clostridium perfringens NetB proteins in Montanide IMS adjuvant increases protective immunity against experimentally-induced necrotic enteritis.

    PubMed

    Lillehoj, Hyun Soon; Jang, Seung Ik; Panebra, Alfredo; Lillehoj, Erik Peter; Dupuis, Laurent; Ben Arous, Juliette; Lee, Seung Kyoo; Oh, Sung Taek

    2017-10-01

    The effects of vaccinating 18-day-old chicken embryos with the combination of recombinant Eimeria profilin plus Clostridium perfringens (C. perfringens) NetB proteins mixed in the Montanide IMS adjuvant on the chicken immune response to necrotic enteritis (NE) were investigated using an Eimeria maxima (E. maxima)/C. perfringens co-infection NE disease model that we previously developed. Eighteen-day-old broiler embryos were injected with 100 μL of phosphate-buffered saline, profilin, profilin plus necrotic enteritis B-like (NetB), profilin plus NetB/Montanide adjuvant (IMS 106), and profilin plus Net-B/Montanide adjuvant (IMS 101). After post-hatch birds were challenged with our NE experimental disease model, body weights, intestinal lesions, serum antibody levels to NetB, and proinflammatory cytokine and chemokine mRNA levels in intestinal intraepithelial lymphocytes were measured. Chickens in ovo vaccinated with recombinant profilin plus NetB proteins/IMS106 and recombinant profilin plus NetB proteins/IMS101 showed significantly increased body weight gains and reduced gut damages compared with the profilin-only group, respectively. Greater antibody response to NetB toxin were observed in the profilin plus NetB/IMS 106, and profilin plus NetB/IMS 101 groups compared with the other three vaccine/adjuvant groups. Finally, diminished levels of transcripts encoding for proinflammatory cytokines such as lipopolysaccharide-induced tumor necrosis factor-α factor, tumor necrosis factor superfamily 15, and interleukin-8 were observed in the intestinal lymphocytes of chickens in ovo injected with profilin plus NetB toxin in combination with IMS 106, and profilin plus NetB toxin in combination with IMS 101 compared with profilin protein alone bird. These results suggest that the Montanide IMS adjuvants potentiate host immunity to experimentally-induced avian NE when administered in ovo in conjunction with the profilin and NetB proteins, and may reduce disease pathology by

  2. Eimeria tenella (Gallus domesticus)

    PubMed Central

    Pierce, A. E.

    1962-01-01

    Complete immunity to a challenge dose of 100,000 sporulated oocysts of Eimeria tenella was developed in fowls 14 days after they had received the last of three graded doses of oocysts of this species, whereas uninfected fowls of comparable age were fully susceptible. In fowls similarly immunized, no detectable first-generation schizogony developed from challenge doses of 10 million oocysts administered to each fowl 21 days after the last of the three graded doses had been administered. Precipitating antibodies were demonstrated in some but not all of these immune fowls by the agar-gel diffusion technique. Precipitin bands, developed as the result of infection, showed a reaction of identity with those induced by parenteral injection of schizont antigen and most of the bands appeared to be directed against protein antigens. Cross reactions were observed between E. tenella antiserum and antigens prepared from the species of coccidia, E. tenella from the fowl and the species E. stiedae from the rabbit. Electrophoretic analysis of serum from immune birds showed an albumin component and four globulin fractions (I-IV); antibody activity was confined to the fraction with the slowest mobility (IV). No significant differences were shown between the electrophoretic analyses at comparable ages of serum from the infected and control groups of fowls between 7 and 63 days of age. The components in both groups altered significantly with time, showing a general rise in protein concentrations. Infected fowls repeatedly showed numerous pyroninophilic cells in the gut mucosa and cells closely resembling globular leucocytes in the deep glands of the caeca. ImagesFIG. 1FIG. 2FIG. 3FIG. 4FIG. 5FIG. 6FIG. 7FIG. 8FIG. 9FIG. 10FIG. 11FIG. 12 PMID:14486445

  3. A comparative transcriptome analysis reveals expression profiles conserved across three Eimeria spp. of domestic fowl and associated with multiple developmental stages.

    PubMed

    Novaes, Jeniffer; Rangel, Luiz Thibério L D; Ferro, Milene; Abe, Ricardo Y; Manha, Alessandra P S; de Mello, Joana C M; Varuzza, Leonardo; Durham, Alan M; Madeira, Alda Maria B N; Gruber, Arthur

    2012-01-01

    Coccidiosis of the domestic fowl is a worldwide disease caused by seven species of protozoan parasites of the genus Eimeria. The genome of the model species, Eimeria tenella, presents a complexity of 55-60MB distributed in 14 chromosomes. Relatively few studies have been undertaken to unravel the complexity of the transcriptome of Eimeria parasites. We report here the generation of more than 45,000 open reading frame expressed sequence tag (ORESTES) cDNA reads of E. tenella, Eimeria maxima and Eimeria acervulina, covering several developmental stages: unsporulated oocysts, sporoblastic oocysts, sporulated oocysts, sporozoites and second generation merozoites. All reads were assembled to constitute gene indices and submitted to a comprehensive functional annotation pipeline. In the case of E. tenella, we also incorporated publicly available ESTs to generate an integrated body of information. Orthology analyses have identified genes conserved across different apicomplexan parasites, as well as genes restricted to the genus Eimeria. Digital expression profiles obtained from ORESTES/EST countings, submitted to clustering analyses, revealed a high conservation pattern across the three Eimeria spp. Distance trees showed that unsporulated and sporoblastic oocysts constitute a distinct clade in all species, with sporulated oocysts forming a more external branch. This latter stage also shows a close relationship with sporozoites, whereas first and second generation merozoites are more closely related to each other than to sporozoites. The profiles were unambiguously associated with the distinct developmental stages and strongly correlated with the order of the stages in the parasite life cycle. Finally, we present The Eimeria Transcript Database (http://www.coccidia.icb.usp.br/eimeriatdb), a website that provides open access to all sequencing data, annotation and comparative analysis. We expect this repository to represent a useful resource to the Eimeria scientific

  4. Identification of Lead Compounds Targeting the Cathepsin B-Like Enzyme of Eimeria tenella

    PubMed Central

    Schaeffer, Marie; Schroeder, Joerg; Heckeroth, Anja R.; Noack, Sandra; Gassel, Michael; Mottram, Jeremy C.

    2012-01-01

    Cysteine peptidases have been implicated in the development and pathogenesis of Eimeria. We have identified a single-copy cathepsin B-like cysteine peptidase gene in the genome database of Eimeria tenella (EtCatB). Molecular modeling of the predicted protein suggested that it differs significantly from host enzymes and could be a good drug target. EtCatB was expressed and secreted as a soluble, active, glycosylated mature enzyme from Pichia pastoris. Biochemical characterization of the recombinant enzyme confirmed that it is cathepsin B-like. Screening of a focused library against the enzyme identified three inhibitors (a nitrile, a thiosemicarbazone, and an oxazolone) that can be used as leads for novel drug discovery against Eimeria. The oxazolone scaffold is a novel cysteine peptidase inhibitor; it may thus find widespread use. PMID:22143531

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

    PubMed

    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.

  6. Higher incidence of Eimeria spp. field isolates sensitive for diclazuril and monensin associated with the use of live coccidiosis vaccination with paracox-5 in broiler farms.

    PubMed

    Peek, H W; Landman, W J M

    2006-09-01

    Twenty European Eimeria spp. field isolates were subjected to an anticoccidial sensitivity test (AST). The anticoccidial drugs tested were diclazuril (Clinacox) and monensin (Elancoban). The assay was performed in a battery cage trial. Infected medicated birds were compared with an unmedicated control group. Coccidial lesion scores and oocyst shedding were used as parameters. The results of the AST show that resistance is common amongst coccidiosis field isolates, especially Eimeria acervulina (68% and 53% resistance for diclazuril and monensin, respectively). Resistance is less frequent amongst Eimeria maxima (38% and 50% resistance for diclazuril and monensin, respectively) and Eimeria tenella isolates (23% and 38% resistance for diclazuril and monensin, respectively). A highly significant influence of the coccidiosis prevention program (live coccidiosis vaccination with Paracox-5 vs. anticoccidial drugs in feed) on the sensitivity patterns of Eimeria spp. field isolates for both diclazuril (P= 0.000) and monensin (P= 0.001) was found. Further, when looking at the single species and each anticoccidial drug level, significantly more sensitivity of E. acervulina for monensin (P= 0.018), E. maxima for diclazuril (P = 0.009), and E. tenella for diclazuril (P = 0.007) was found in isolates originating from vaccinated flocks. Moreover, for E. acervulina and diclazuril, E. maxima and monensin, and E. tenella and monensin a trend toward higher sensitivity of isolates for these products was found when live coccidiosis vaccination was applied. The present study shows that sensitivity for the anticoccidial drugs diclazuril and monensin is more frequent in Eimeria spp. field isolates originating from broiler farms where a coccidiosis vaccination policy is followed.

  7. Downregulation of Chicken Interleukin-17 Receptor A during Eimeria Infection

    PubMed Central

    Kim, Woo H.; Jeong, Jipseol; Park, Ae R.; Yim, Dongjean; Kim, Suk; Chang, Hong H.; Yang, Seung-Hak; Kim, Dong-Hee; Lillehoj, Hyun S.

    2014-01-01

    Both interleukin-17A (IL-17A) and IL-17F are proinflammatory cytokines that have an important role in intestinal homeostasis via receptor signaling. These cytokines have been characterized in chickens, but very little is known about their receptors and their functional activity. We provide here the first description of the sequence analysis, bioactivity, and comparative expression analysis of chicken IL-17RA (chIL-17RA) in chickens infected with Salmonella and Eimeria, two major infectious agents of gastrointestinal diseases of poultry of economic importance. A full-length chIL-17RA cDNA with a 2,568-bp coding region was identified from chicken thymus cDNA. chIL-17RA shares ca. 46% identity with mammalian homologues and 29.2 to 31.5% identity with its piscine counterparts. chIL-17RA transcript expression was relatively high in the thymus and in the chicken macrophage cell line HD11. The chIL-17RA-specific small interfering RNA inhibits interleukin-6 (IL-6), IL-8, and IL-1β mRNA expression in chicken embryo fibroblast cells (but not in DF-1 cells) stimulated with chIL-17A or chIL-17F. Interaction between chIL-17RA and chIL-17A was confirmed by coimmunoprecipitation. Downregulation of chIL-17RA occurred in concanavalin A- or lipopolysaccharide-activated splenic lymphocytes but not in poly(I·C)-activated splenic lymphocytes. In Salmonella- and Eimeria-infected chickens, the expression levels of the chIL-17RA transcript were downregulated in intestinal tissues from chickens infected with two Eimeria species, E. tenella or E. maxima, that preferentially infect the cecum and jejunum, respectively. However, chIL-17RA expression was generally unchanged in Salmonella infection. These results suggest that chIL-17RA has an important role in mucosal immunity to intestinal intracellular parasite infections such as Eimeria infection. PMID:24980970

  8. Downregulation of chicken interleukin-17 receptor A during Eimeria infection.

    PubMed

    Kim, Woo H; Jeong, Jipseol; Park, Ae R; Yim, Dongjean; Kim, Suk; Chang, Hong H; Yang, Seung-Hak; Kim, Dong-Hee; Lillehoj, Hyun S; Min, Wongi

    2014-09-01

    Both interleukin-17A (IL-17A) and IL-17F are proinflammatory cytokines that have an important role in intestinal homeostasis via receptor signaling. These cytokines have been characterized in chickens, but very little is known about their receptors and their functional activity. We provide here the first description of the sequence analysis, bioactivity, and comparative expression analysis of chicken IL-17RA (chIL-17RA) in chickens infected with Salmonella and Eimeria, two major infectious agents of gastrointestinal diseases of poultry of economic importance. A full-length chIL-17RA cDNA with a 2,568-bp coding region was identified from chicken thymus cDNA. chIL-17RA shares ca. 46% identity with mammalian homologues and 29.2 to 31.5% identity with its piscine counterparts. chIL-17RA transcript expression was relatively high in the thymus and in the chicken macrophage cell line HD11. The chIL-17RA-specific small interfering RNA inhibits interleukin-6 (IL-6), IL-8, and IL-1β mRNA expression in chicken embryo fibroblast cells (but not in DF-1 cells) stimulated with chIL-17A or chIL-17F. Interaction between chIL-17RA and chIL-17A was confirmed by coimmunoprecipitation. Downregulation of chIL-17RA occurred in concanavalin A- or lipopolysaccharide-activated splenic lymphocytes but not in poly(I·C)-activated splenic lymphocytes. In Salmonella- and Eimeria-infected chickens, the expression levels of the chIL-17RA transcript were downregulated in intestinal tissues from chickens infected with two Eimeria species, E. tenella or E. maxima, that preferentially infect the cecum and jejunum, respectively. However, chIL-17RA expression was generally unchanged in Salmonella infection. These results suggest that chIL-17RA has an important role in mucosal immunity to intestinal intracellular parasite infections such as Eimeria infection.

  9. Montanide™ ISA 71 VG adjuvant enhances antibody and cell-mediated immune responses to profilin subunit antigen vaccination and promotes protection against Eimeria acervulina and Eimeria tenella.

    PubMed

    Jang, Seung I; Lillehoj, Hyun S; Lee, Sung Hyen; Lee, Kyung Woo; Lillehoj, Erik P; Bertrand, François; Dupuis, Laurent; Deville, Sébastien

    2011-01-01

    The present study was conducted to investigate the immunoenhancing effects of Montanide™ ISA 71 VG adjuvant on profilin subunit antigen vaccination. Broiler chickens were immunized subcutaneously with a purified Eimeria acervulina recombinant profilin protein, either alone or mixed with ISA 71 VG, and host immune responses were evaluated. After secondary immunization, antigen-specific antibody and T-cell responses were higher in the group which received profilin plus ISA 71 VG compared with the other groups. Furthermore, body weight gains and fecal oocyst shedding were evaluated following oral challenge infection with live E. acervulina or Eimeria tenella oocysts. Vaccination with profilin plus ISA 71 VG reduced oocyst shedding compared with animals immunized with profilin alone. These results demonstrate that the recombinant profilin subunit vaccine, when given in combination with Montanide™ ISA 71 VG, augments protective immunity against E. acervulina and E. tenella. Published by Elsevier Inc.

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

    SciTech Connect

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

    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.

  11. Immunization of broiler chicks by in ovo injection of infective stages of Eimeria.

    PubMed

    Weber, F H; Genteman, K C; LeMay, M A; Lewis, D O; Evans, N A

    2004-03-01

    Immunization of chickens by in ovo injection of infective stages of 5 species of Eimeria was investigated. Fertile Hubbard x Petersen broiler chicken eggs were injected through the air cell on d 18 of incubation with oocysts of E. acervulina, E. maxima, E. mitis, E. praecox, or E. brunetti. Injected doses of all species ranged from 1 x 10(2) to 1 x 10(6) sporulated oocysts per egg. Chicks receiving oocysts in ovo shed oocysts posthatch. After 2 wk in wire-floored cages, birds were given a challenge infection with the homologous Eimeria species. Chicks immunized by in ovo injection of oocysts had significantly reduced lesion scores, improved weight gain, or reduced oocyst output compared with their nonimmunized counterparts. In additional studies, eggs were injected with 1 x 10(5) sporozoites of E. tenella, E. maxima, or E. acervulina per egg. Sporozoites of E. acervulina were not infective for chick embryos when administered in phosphate-buffered saline, but if sporozoites were suspended in tissue culture medium when injected in ovo, hatched chicks shed oocysts with peak output occurring 3 to 4 d posthatch. Sporozoites of E. maxima and E. tenella were infective for 18-d-old embryos regardless of the vehicle. The results demonstrate that immunization of broiler chickens against several species of coccidia by in ovo injection of oocysts is feasible. The infectivity of sporozoites for 18-d-old chick embryos varied depending on the species of Eimeria and the vehicle in which the sporozoites were suspended prior to injection.

  12. MAXIMA: Observations of CMB anisotropy

    NASA Astrophysics Data System (ADS)

    Rabii, Bahman

    This document describes the Millimeter Anisotropy eXperiment IMaging Array (MAXIMA), a balloon-borne experiment measuring the temperature anisotropy of the Cosmic Microwave Background (CMB) on angular scales of 10 ' to 5°. MAXIMA data are used to discriminate between cosmological models and to determine cosmological parameters. MAXIMA maps the CMB using 16 bolometric detectors observing in spectral bands centered at 150 GHz, 230 GHz, and 410 GHz, with 10 ' resolution at all frequencies. The combined receiver sensitivity to CMB anisotropy is ˜40 μK sec , the best reported by any CMB experiment. Systematic errors are rejected by using four uncorrelated spatial modulations, multiple independent CMB observations, heavily baffled optics, and strong spectral discrimination. Observation patterns are well cross-linked and optimized for the extraction of cosmological information. Pointing is reconstructed to an accuracy of 1'. Absolute calibration uncertainty of 3 4% is the best achieved by any sub-orbital CMB experiment. Two MAXIMA flights were launched from the National Scientific Balloon Facility in Palestine Texas in 1998 and 1999. During a total of 8.5 hours of CMB observations, 300 deg2 of the sky were mapped, with ˜50 deg2 overlap between the two flights. The observed region was selected for low foreground emission and post-flight data analysis confirms that foreground contamination is negligible. Cosmological results are presented from the 1998 flight, MAXIMA-I , in which 122 deg2 of sky were mapped over 3 hours. A maximum likelihood map with 3' pixelization is obtained from the three most sensitive and best tested detectors. The angular power spectrum derived from this map shows a narrow peak near ℓ = 200, and is consistent with inflationary Big Bang models. Within these models, cosmological parameters are estimated, including total density Ωtot = 0.9+0.18-0.16 , baryon density Ωbh2 = 0.033 ± 0.013, and power spectrum normalization C

  13. The use of dual-energy X-ray absorptiometry to assess the impact of Eimeria infections in broiler chicks.

    PubMed

    Fetterer, Raymond H; Miska, Katarzyna B; Mitchell, Alva D; Jenkins, Mark C

    2013-06-01

    A number of parameters have been used to assess the impact ofcoccidiosis on chickens in clinical settings as well as in experimental studies. However, a rapid way to determine body composition would be useful to evaluate or compare responses to coccidia and could give further insight into the metabolic impact of infection. The current study evaluates the use of dual X-ray absorptiometry (DEXA) to determine the impact of coccidiosis on body composition in chicks receiving inoculations with single or mixed species of Eimeria. Chicks infected with Eimeria maxima, Eimeria acervulina, or Eimeria tenella had altered parameters of body composition as measured by DEXA at 6 days postinfection (PI). The greatest effects were noted in birds infected with E. acervulina or E. maxima, where lean mass and fat were reduced from control values about 75% and 85%, respectively. In chicks infected with E. tenella, tissue and fat were reduced about 10%. Bone mineral content (BMC) was about 75% of control values in birds infected with E. acervulina or E. maxima, but only E. acervulina altered bone mineral density (BMD). The decreases in BMC and BMD are likely due to malabsorption. In chicks receiving a mixed coccidian infection, all DEXA parameters were significantly decreased at 8 days PI compared with age-matched controls. As with single infections, BMD and BMC were significantly depressed (P < 0.05). Values of all DEXA parameters were near 92% of control values by day 16 PI. Analysis of all birds in the current study indicates DEXA tissue weight slightly underestimated the gravimetrically measured weight by about 3%. The current results demonstrate that DEXA is a potentially important tool for the rapid evaluation of the effect of coccidiosis on broiler chicks and suggest it can be useful for evaluation of vaccines and other disease controls.

  14. Montanide™ ISA 71 VG adjuvant enhances antibody and cell-mediated immune responses to profilin subunit antigen vaccination and promotes protection against Eimeria acervulina and Eimeria tenella. Experimental Parasitology

    USDA-ARS?s Scientific Manuscript database

    The present study was conducted to investigate the immunoenhancing effects of MontanideTM ISA 71 VG adjuvant on profilin subunit antigen vaccination. Broiler chickens were immunized subcutaneously with a purified Eimeria acervulina recombinant profilin protein, either alone or mixed with ISA 71 VG, ...

  15. Evaluation of novel adjuvant Eimeria profilin complex on intestinal host immune responses against live E. acervulina challenge infection

    USDA-ARS?s Scientific Manuscript database

    The effects of two novel adjuvants, QCDC (Quil A/cholesterol/DDA/Carbopol) and QCDCRT (QCDC/Bay R1005/cytosine-phosphate-guanosine oligodeoxynucleotides, CpG) emulsified with profilin, a conserved Eimeria recombinant protein, against avian coccidiosis were determined in broiler chickens. Chickens we...

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

    NASA Astrophysics Data System (ADS)

    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.

  17. Efficacy of diclazuril in comparison with chemical and ionophorous anticoccidials against Eimeria spp. in broiler chickens in floor pens.

    PubMed

    Conway, D P; Mathis, G F; Johnson, J; Schwartz, M; Baldwin, C

    2001-04-01

    Two 42-d floor pen studies were conducted with commercial broiler chickens to measure the efficacy of 1 ppm diclazuril in the starter or grower diet in shuttle programs with 66 ppm salinomycin. Study 1 compared a salinomycin to diclazuril (starter to grower diet) shuttle treatment with salinomycin to salinomycin, salinomycin to 100 ppm monensin, salinomycin to 99.8 ppm lasalocid, and unmedicated treatments. Study 2 compared a diclazuril to salinomycin (starter to grower) shuttle treatment with 125 ppm nicarbazin to salinomycin, 79.2 ppm narasin + nicarbazin to salinomycin, 125 ppm zoalene to salinomycin, and unmedicated treatments. Fifty 1-d-old chicks were randomly allotted to each of 10 pens per treatment in each study using a randomized complete block design. Starter (Days 0 to 21) and grower (Days 22 to 37) diets in each study contained 55 ppm bacitracin methylene disalicylate. The finisher diet (Days 38 to 42) in each study was unmedicated. Birds were inoculated via their feed on Day 22 (Study 1) or Day 15 (Study 2) with a mixed inoculum of Eimeria acervulina, Eimeria maxima, and Eimeria tenella. Four birds per pen (two male and two female) were randomly selected in each study for coccidial lesion scores on Day 6 postinoculation. These studies demonstrated that the use of 1 ppm diclazuril in shuttle programs was highly efficacious against a mixed inoculum of Eimeria spp. in comparison with nicarbazin, narasin + nicarbazin, and zoalene in starter diets and salinomycin, monensin, and lasalocid in grower diets.

  18. Bovine Eimeria species in Austria.

    PubMed

    Koutny, H; Joachim, A; Tichy, A; Baumgartner, W

    2012-05-01

    Bovine eimeriosis is considered to be of considerable importance for the productivity and health of cattle worldwide. Despite the importance of cattle farming in Austria, little is known in this country about the abundance and distribution of bovine Eimeria spp. The objective of this study was to obtain detailed information about the occurrence of different Eimeria spp. on Austrian dairy farms. Fecal samples from individual calves (n = 868) from 296 farms all over Austria (82 districts) were collected. Additionally, each farmer was questioned about the occurrence of calf diarrhea, and about the knowledge on coccidiosis and possible control measures. On 97.97% of the investigated farms, calves excreted Eimeria oocysts, and 83.67% of the individual samples were positive. After sporulation of positive samples pooled from each farm, 11 Eimeria species were found, with E. bovis (in 65.54% of the samples and 27.74% of the farms), E.zuernii (63.85%/13.86%), E. auburnensis (56.76%/13.41%) and E. ellipsoidalis (54.05%/14.38%) being the most prevalent, followed by E. alabamensis (45.61%/11.56%), E. subspherica (35.14%/5.5.05%), E. cylindrica (33.11%/7.00%), and E. canadensis (31.08%/7.74%). E. wyomingensis, E. pellita and E. bukidnonensis were only found sporadically (3.04-4.73% of the samples and 0.16-0.59% of the farms). Mixed infections were present on all farms (2-9 Eimeria species/farm). Prevalences by state provinces were high throughout with 77.1-87.9% of the samples and 93.8-100% of the farms. Lower Austria had the highest percentage of positive farms, and Vorarlberg the lowest. Individual OPG (oocysts per gram of feces) values were generally low; 75% of the samples had an OPG of 1,000 or less. The highest detected OPG was 72,400. The mean OPG was 2,525 with above average numbers in Tirol, Carinthia, and Lower Austria. The mean OPG values were significantly positively correlated with the cattle density in the different districts. The majority of the samples were from

  19. Molecular characterization of Eimeria species in macropods.

    PubMed

    Yang, Rongchang; Fenwick, Stan; Potter, Abbey; Elliot, Aileen; Power, Michelle; Beveridge, Ian; Ryan, Una

    2012-10-01

    A total of 597 faecal samples were collected from western grey kangaroos (Macropus fuliginosus), Euros (M. robustus), red kangaroos (M. rufus) in Western Australia and Eastern Grey Kangaroos (M. giganteus) from Victoria and screened for the presence of Eimeria by PCR at the 18S ribosomal RNA (rRNA) locus. The overall prevalence was 24.3% (145/597). At the 18S rRNA locus, sequences were obtained for 25 of the 145 positives. Phylogenetic analysis indicated that all the macropod-derived Eimeria species grouped in a separate marsupial clade that included Eimeria trichosuri from brushtail possums. At least 6 different clades were identified within the marsupial isolates and many of the genotypes identified are likely to be valid species, however morphological and biological data need to be collected to match sequences to previously characterized Eimeria species or identify if they are new species.

  20. Protective effects of Aloe vera-based diets in Eimeria maxima-infected broiler chickens

    USDA-ARS?s Scientific Manuscript database

    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...

  1. Scaling exponents for ordered maxima

    SciTech Connect

    Ben-Naim, E.; Krapivsky, P. L.; Lemons, N. W.

    2015-12-22

    We study extreme value statistics of multiple sequences of random variables. For each sequence with N variables, independently drawn from the same distribution, the running maximum is defined as the largest variable to date. We compare the running maxima of m independent sequences and investigate the probability SN that the maxima are perfectly ordered, that is, the running maximum of the first sequence is always larger than that of the second sequence, which is always larger than the running maximum of the third sequence, and so on. The probability SN is universal: it does not depend on the distribution from which the random variables are drawn. For two sequences, SN~N–1/2, and in general, the decay is algebraic, SN~N–σm, for large N. We analytically obtain the exponent σ3≅1.302931 as root of a transcendental equation. Moreover, the exponents σm grow with m, and we show that σm~m for large m.

  2. Scaling exponents for ordered maxima

    DOE PAGES

    Ben-Naim, E.; Krapivsky, P. L.; Lemons, N. W.

    2015-12-22

    We study extreme value statistics of multiple sequences of random variables. For each sequence with N variables, independently drawn from the same distribution, the running maximum is defined as the largest variable to date. We compare the running maxima of m independent sequences and investigate the probability SN that the maxima are perfectly ordered, that is, the running maximum of the first sequence is always larger than that of the second sequence, which is always larger than the running maximum of the third sequence, and so on. The probability SN is universal: it does not depend on the distribution frommore » which the random variables are drawn. For two sequences, SN~N–1/2, and in general, the decay is algebraic, SN~N–σm, for large N. We analytically obtain the exponent σ3≅1.302931 as root of a transcendental equation. Moreover, the exponents σm grow with m, and we show that σm~m for large m.« less

  3. A comparison of Eimeria oocysts isolated from litter and fecal samples from broiler houses at two farms with different management schemes during one growout.

    PubMed

    Stayer, P A; Pote, L M; Keirs, R W

    1995-01-01

    Eimeria oocyst populations were monitored in broiler houses on two farms during a growout and prior to the placement of new chicks on the same litter. Numbers of oocysts in litter were enumerated at Weeks 0, 5, 6, 7, and 9 and in feces at Weeks 5,6, and 7 in broiler houses with different management practices (both farms had identical coccidiostat programs but Farm A had a history of poor performance whereas Farm B had excellent performance records). On Farm A the number of oocysts in litter increased (P < .05) at the time of slaughter (Week 7) and placement of a subsequent flock (Week 9) when compared with Day 0. The number of oocysts in feces on Farm A also increased (P < .05) from Week 5 to 7. On Farm B there were no differences in numbers of oocysts in litter or feces over time. Farm B had (P < .05) lower numbers of oocysts in litter than Farm A at Week 7. The number of oocysts in feces from Farm B were consistently lower (P < .05) than Farm A at Weeks 5, 6, and 7. There were no differences in the species of Eimeria in litter between farms; however, there were differences in the species composition over time. Small (Eimeria mitis) and medium (primarily Eimeria acervulina) oocysts increased significantly over time, whereas the large oocysts (Eimeria tenella and Eimeria brunetti) and Eimeria maxima did not change over time. An increase (P < .05) in large oocysts was observed in fecal samples from Farm A over time.

  4. Effects of in ovo vaccination and anticoccidials on the distribution of Eimeria spp. in poultry litter and serum antibody titers against coccidia in broiler chickens raised on the used litters.

    PubMed

    Lee, Kyung Woo; Lillehoj, Hyun S; Jang, Seung I; Pagès, Marc; Bautista, Daniel A; Pope, Conrad R; Ritter, G Donald; Lillehoj, Erik P; Neumann, Anthony P; Siragusa, Gregory R

    2012-08-01

    The present study reports the effects of various field anticoccidial programs on the distribution of Eimeria spp. in poultry litter and serum antibody titers against coccidia in broiler chickens raised on the used litters. The programs included in ovo vaccination and various medications with either chemicals, ionophores, or both. In general, serum samples from these chickens showed anticoccidial antibody titers when tested at days 7 and 14 post hatch with the peak response at day 43. Serum anticoccidial titers were highest in birds fed a non-medicated diet compared with those vaccinated or fed medicated diets. Total number of Eimeria oocysts and the composition of Eimeria spp. present in the litter samples from different treatment groups varied depending on the type of anticoccidial program. Oocyst counts in general ranged from 3.7×10(3) to 7.0×10(4) per g of litter. Importantly, both morphological and molecular typing studies revealed four major predominant Eimeria spp., E. acervulina, E. maxima, E. praecox, and E. tenella in the litter samples. Collectively, these results indicate that the field anticoccidial programs influenced the type and abundance of Eimeria spp. present in the litter samples and also modulated host immune response to Eimeria.

  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.

    PubMed

    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. A conserved 19-kDa Eimeria tenella antigen is a profilin-like protein.

    PubMed

    Fetterer, R H; Miska, K B; Jenkins, M C; Barfield, R C

    2004-12-01

    A wide range of recombinant proteins from Eimeria species have been reported to offer some degree of protection against infection and disease, but the specific biological function of these proteins is largely unknown. Previous studies have demonstrated a 19-kDa protein of unknown function designated SZ-1 in sporozoites and merozoites of Eimeria acervulina that can be used to confer partial protection against coccidiosis. Reverse transcriptase-polymerase chain reaction indicated that the gene for SZ-1 is expressed by all the asexual stages of Eimeria tenella. Rabbit antisera to recombinant SZ-1 recognized an approximately 19-kDa protein from extracts of E. tenella sporozoites, merozoites, sporulated oocysts, and oocysts in various stages of sporulation. Immunofluorescence antibody staining indicated specific staining of E. tenella sporozoites and merozoites. Staining was most intense in the cytoplasm of the posterior end of the parasite. The primary amino acid sequence of the gene for E. tenella SZ-1 deduced from the E. tenella genome indicated a conserved domain for the actin-regulatory protein profilin. A conserved binding site for poly-L-proline (PLP), characteristic of profilin was also observed. SZ-1 was separated from soluble extract of E. tenella proteins by affinity chromatography using a PLP ligand, confirming the ability of SZ-1 to bind PLP. SZ-1 also partially inhibited the polymerization of actin. The current results are consistent with the classification of SZ-1 as a profilin-related protein.

  7. Eimeria infections in goats in Southern Portugal.

    PubMed

    Silva, Liliana Machado Ribeiro da; Vila-Viçosa, Maria João Martins; Nunes, Telmo; Taubert, Anja; Hermosilla, Carlos; Cortes, Helder Carola Espiguinha

    2014-01-01

    Coccidiosis caused by Eimeria species is a major form of intestinal infection affecting intensively and semi-intensively reared goats. The province of Alentejo is the main goat-producing area in Portugal. Therefore, all 15 Serpentina goat farms in Alentejo were analyzed regarding the occurrence and diversity of Eimeria species. Fecal samples obtained from 144 animals (52.1% dairy goats, 47.9% pre-pubertal goats) were examined using the modified McMaster technique to determine the number of oocysts per gram of feces. Eimeria spp. oocysts were present in 98.61% of the fecal samples and, overall, nine different Eimeria species were identified. The most prevalent species were E. ninakohlyakimovae (88%) and E. arloingi (85%), followed by E. alijevi (63%) and E. caprovina (63%). The average number of oocysts shed was significantly lower in dairy goats than in pre-adult animals. Astonishingly, no clinical signs of coccidiosis were observed in any of the animals examined, even though they were shedding high numbers of oocysts and were infected with highly pathogenic species. Thus, implementation of routine diagnostic investigation of the occurrence and diversity of caprine Eimeria species may be a useful tool for determination and better understanding of their potential economic impact on goat herds in southern Portugal.

  8. Using Maxima in the Mathematics Classroom

    ERIC Educational Resources Information Center

    Fedriani, Eugenio M.; Moyano, Rafael

    2011-01-01

    Coming from the Macsyma system and adapted to the Common Lisp standard, Maxima can be regarded as a tool for a frequent use in the mathematics classroom. The main aim of this work is to show some possibilities of Maxima and its graphical interface through our experience as Mathematics teachers in Business degrees, although it can be easily spread…

  9. T lymphocyte roles during Eimeria acervulina and Eimeria tenella infections.

    PubMed

    Trout, J M; Lillehoj, H S

    1996-09-01

    This study evaluated the effects of selective depletion of T lymphocytes on Eimeria infections in chickens. Cell depletions were initiated in day- or week-old Hyline SC strain chickens using intra-peritoneal injections of monoclonal antibodies to CD4, CD8, or T cell receptor (TCR) alpha/beta. Control chickens received injections of irrelevant monoclonal antibody or phosphate buffered saline (PBS). Following the establishment of cell depletion, chickens were infected orally with E. acervulina or E. tenella, 1 x 10(4) oocysts for primary infections and 2 x 10(5) oocysts for secondary infections. Chickens treated with anti CD4 monoclonal antibody produced significantly more oocysts than controls following primary E. tenella but not E. acervulina infections. Development of resistance to challenge infection was unaffected. These results suggest that CD4+ lymphocytes are important in controlling primary infection with E. tenella. Chickens treated with anti-CD8 or anti-TCR alpha/beta monoclonal antibodies produced significantly fewer oocysts than controls following primary infection but significantly more oocysts than controls following secondary infection with both E. tenella and E. acervulina. Additionally, anti-CD8 treatment abrogated resistance to challenge infection. CD8-depleted chickens may exhibit decreased oocyst production following primary infection due to a lack of CD8+ lymphocytes to serve as transporting cells for sporozoites. The abrogation of resistance to secondary infection in CD8- and TCR alpha/beta-depleted chickens suggests that these cells are necessary for the development of protective immunity to coccidia.

  10. The role of sialyl glycan recognition in host tissue tropism of the avian parasite Eimeria tenella.

    PubMed

    Lai, Livia; Bumstead, Janene; Liu, Yan; Garnett, James; Campanero-Rhodes, Maria A; Blake, Damer P; Palma, Angelina S; Chai, Wengang; Ferguson, David J P; Simpson, Peter; Feizi, Ten; Tomley, Fiona M; Matthews, Stephen

    2011-10-01

    Eimeria spp. are a highly successful group of intracellular protozoan parasites that develop within intestinal epithelial cells of poultry, causing coccidiosis. As a result of resistance against anticoccidial drugs and the expense of manufacturing live vaccines, it is necessary to understand the relationship between Eimeria and its host more deeply, with a view to developing recombinant vaccines. Eimeria possesses a family of microneme lectins (MICs) that contain microneme adhesive repeat regions (MARR). We show that the major MARR protein from Eimeria tenella, EtMIC3, is deployed at the parasite-host interface during the early stages of invasion. EtMIC3 consists of seven tandem MAR1-type domains, which possess a high specificity for sialylated glycans as shown by cell-based assays and carbohydrate microarray analyses. The restricted tissue staining pattern observed for EtMIC3 in the chicken caecal epithelium indicates that EtMIC3 contributes to guiding the parasite to the site of invasion in the chicken gut. The microarray analyses also reveal a lack of recognition of glycan sequences terminating in the N-glycolyl form of sialic acid by EtMIC3. Thus the parasite is well adapted to the avian host which lacks N-glycolyl neuraminic acid. We provide new structural insight into the MAR1 family of domains and reveal the atomic resolution basis for the sialic acid-based carbohydrate recognition. Finally, a preliminary chicken immunization trial provides evidence that recombinant EtMIC3 protein and EtMIC3 DNA are effective vaccine candidates.

  11. Identification and molecular characterization of microneme 5 of Eimeria acervulina.

    PubMed

    Zhang, ZhenChao; Huang, JingWei; Li, MengHui; Sui, YuXia; Wang, Shuai; Liu, LianRui; Xu, LiXin; Yan, RuoFeng; Song, XiaoKai; Li, XiangRui

    2014-01-01

    In the present study, the microneme 5 gene of Eimeria acervulina (E. acervulina) (EaMIC5) was cloned and characterized. Specific primers for the rapid amplification of cDNA ends (RACE) were designed based on the expressed sequence tag (EST, GenBank Accession No. EH386430.1) to amplify the 3'- and 5'-ends of EaMIC5. The full length cDNA of this gene was obtained by overlapping the sequences of 3'- and 5'-extremities and amplification by reverse transcription PCR. Sequence analysis revealed that the open reading frame (ORF) of EaMIC5 was 336 bp and encoded a protein of 111 amino acids with 12.18 kDa. The ORF was inserted into pET-32a (+) to produce recombinant EaMIC5. Using western blotting assay, the recombinant protein was successfully recognized by the sera of chicks experimentally infected with E. acervulina, while the native protein in the somatic extract of sporozoites was as well detected by sera from rats immunized with the recombinant protein of EaMIC5. Immunofluorescence analysis using antibody against recombinant protein EaMIC5 indicated that this protein was expressed in the sporozoites and merozoites stages of E. acervulina. Animal challenge experiments demonstrated that the recombinant protein of EaMIC5 could significantly increase the average body weight gains, decrease the mean lesion scores and the oocyst outputs of the immunized chickens, and presented anti-coccidial index (ACI) more than 160. All the above results suggested that the EaMIC5 was a novel E. acervulina antigen and could be an effective candidate for the development of a new vaccine against this parasite.

  12. Eimeria species in dairy goats in Brazil.

    PubMed

    Cavalcante, Antônio César Rocha; Teixeira, Marcel; Monteiro, Jomar Patrício; Lopes, Carlos Wilson Gomes

    2012-02-10

    The focus of this work is to determine the distribution and identify species of Eimeria parasites of dairy goats in the livestock of the National Goat and Sheep Research Center in Sobral, State of Ceará, Northeast Brazil. Results showed the presence of multiple species in 196 of 215 analyzed samples (91.2%). Fifty five out of these were from kids (28%) and 141 from adult goats (72%). Eight different Eimeria species were identified and their prevalence in the herd was: Eimeria alijevi Musaev, 1970 (26.7%), E. arloingi (Marotel, 1905) Martin, 1909 (20.6%), E. hirci Chevalier, 1966 (18%), E. ninakohlyakimovae Yakimoff & Rastegaieff, 1930 (16.2%), E. jolchijevi Musaev, 1970 (8.7%), E. christenseni Levine, Ivens & Fritz, 1962 (6%), E. caprovina Lima, 1980 (2.8%) and E. caprina Lima, 1979 (1%). Moreover, E. ninakohlyakimovae showed higher prevalence in kids (97%), followed by E. arloingi and E. alijevi (88%). On the other hand, E. alijevi (77%) was more common in adult goats followed by E. hirci (74%) and E. ninakohlyakimovae (70%). The species E. caprina had low frequency in both kids (27%) and adult goats (13%). Data indicated that infection was relatively common among kids and adult goats. The implementation of a routine diagnostic strategy can be useful in maintaining Eimeria populations under monitoring and will enable the determination of its potential impact on dairy goat herds in Northeast Brazil.

  13. STUDIES ON THE COCCIDIUM, EIMERIA TENELLA,

    DTIC Science & Technology

    Essentially pure sporulated Eimeria tenella oocyst suspensions, suitable for experimental purposes, were obtained by shaking at 30 C homogenized...Ammonium hydroxide, aqueous iodine, and ethylene oxide were the only disinfectants found effective against E. tenella oocysts. Cross-infection studies with other animal species were uniformly unsuccessful. (Author)

  14. Sex and Eimeria: a molecular perspective.

    PubMed

    Walker, Robert A; Ferguson, David J P; Miller, Catherine M D; Smith, Nicholas C

    2013-12-01

    Eimeria is a common genus of apicomplexan parasites that infect diverse vertebrates, most notably poultry, causing serious disease and economic loss. Like all apicomplexans, eimerians have a complex life cycle characterized by asexual divisions that amplify the parasite population in preparation for sexual reproduction. This can be divided into three events: gametocytogenesis, producing gametocytes from merozoites; gametogenesis, producing microgametes and macrogametes from gametocytes; and fertilization of macrogametes by microgametes, producing diploid zygotes with ensuing meiosis completing the sexual phase. Sexual development in Eimeria depends on the differential expression of stage-specific genes, rather than presence or absence of sex chromosomes. Thus, it involves the generation of specific structures and, implicitly, storage of proteins and regulation of protein expression in macrogametes, in preparation for fertilization. In Eimeria, the formation of a unique, resilient structure, the oocyst wall, is essential for completion of the sexual phase and parasite transmission. In this review, we piece together the molecular events that underpin sexual reproduction in Eimeria and use additional details from analogous events in Plasmodium to fill current knowledge gaps. The mechanisms governing sexual stage formation and subsequent fertilization may represent targets for counteracting parasite transmission.

  15. The use of diclazuril in extended withdrawal anticoccidial programs: 1. Efficacy against Eimeria spp. in broiler chickens in floor pens.

    PubMed

    Conway, D P; Mathis, G F; Lang, M

    2002-03-01

    A 49-d floor pen study was conducted with broiler chickens to compare the effects of different anticoccidial withdrawal times on the efficacy of 1 ppm diclazuril. The starter diet in three treatments contained 66 ppm salinomycin + 50 ppm roxarsone (3-nitro-4-hydroxyphenylarsonic acid), followed by 1 ppm diclazuril in the grower diet commencing on Day 17. Diclazuril was withdrawn from these treatments on Day 28, 35, or 42 (finisher diet), respectively. Two other treatments in the study were given 66 ppm salinomycin + 50 ppm roxarsone in the starter and grower diets to Day 28 or no anticoccidial (unmedicated). The starter (Days 0 to 16), grower (Days 17 to 35), and finisher (Days 36 to 49) diets in each treatment included 55 ppm bacitracin methylene disalicylate for growth promotion. Fifty 1-d-old chicks were randomly allotted to each of 10 pens per treatment using a randomized complete block design. Birds in each pen were raised on litter naturally contaminated with a mixture of Eimeria acervulina, Eimeria maxima, and Eimeria tenella. The results demonstrated that some performance loss occurred in the salinomycin Day 28 treatment. Means for weight gain and feed conversion on Day 49 were improved (P < 0.05) in each diclazuril treatment in comparison with the salinomycin and unmedicated treatments. Feed conversion means in the diclazuril Day 35 and Day 42 treatments were improved (P < 0.05) in comparison with the diclazuril Day 28 treatment, indicating that shorter withdrawal times provided further benefit.

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

    PubMed

    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. Eimeria tenella protein trafficking: differential regulation of secretion versus surface tethering during the life cycle.

    PubMed

    Marugan-Hernandez, V; Long, E; Blake, D; Crouch, C; Tomley, F

    2017-07-04

    Eimeria spp. are intracellular parasites that have a major impact on poultry. Effective live vaccines are available and the development of reverse genetic technologies has raised the prospect of using Eimeria spp. as recombinant vectors to express additional immunoprotective antigens. To study the ability of Eimeria to secrete foreign antigens or display them on the surface of the sporozoite, transiently transfected populations of E. tenella expressing the fluorescent protein mCherry, linked to endogenous signal peptide (SP) and glycophosphatidylinositol-anchor (GPI) sequences, were examined. The SP from microneme protein EtMIC2 (SP2) allowed efficient trafficking of mCherry to cytoplasmic vesicles and following the C-terminal addition of a GPI-anchor (from surface antigen EtSAG1) mCherry was expressed on the sporozoite surface. In stable transgenic populations, mCherry fused to SP2 was secreted into the sporocyst cavity of the oocysts and after excystation, secretion was detected in culture supernatants but not into the parasitophorous vacuole after invasion. When the GPI was incorporated, mCherry was observed on the sporozites surface and in the supernatant of invading sporozoites. The proven secretion and surface exposure of mCherry suggests that antigen fusions with SP2 and GPI of EtSAG1 may be promising candidates to examine induction of protective immunity against heterologous pathogens.

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

    PubMed

    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.

  19. Prevalence and drug resistance of avian Eimeria species in broiler chicken farms of Zhejiang province, China.

    PubMed

    Lan, L-H; Sun, B-B; Zuo, B-X-Z; Chen, X-Q; Du, A-F

    2017-07-01

    In this study, coccidia were isolated and identified from 5 main poultry farms located in Zhejiang province, eastern China. The overall prevalence of Eimeria spp. was 30.7% (95 of 310). Five common species were observed: E. tenella, E. acervulina, E. maxima, E. necatrix, and E. mitis. Two isolates (HZ and QZ) were tested for sensitivity to 8 anticoccidial drugs using 4 indexes including anticoccidial index (ACI), percent of optimum anticoccidial activity (POAA), reduction of lesion scores (RLS), and relative oocyst production (ROP): sulfachloropyrazine, toltrazuril, diclazuril, sulfamonomethoxine/trimethoprim, and amprolium; sulfaquinoxaline/sulfadimethoxine, nicarbazin, and halofuginone. The results showed that the 2 isolates have developed various degrees of resistance to most of the tested drugs. The multi-resistance coccidia are a potential threat to local poultry farming. Rotation of anticoccidial drugs and shuttle programs are recommended to prevent further economic losses caused by coccidiosis. © 2017 Poultry Science Association Inc.

  20. Immunogenicity and protective efficacy of an Eimeria vaccine candidate based on Eimeria tenella immune mapped protein 1 and chicken CD40 ligand.

    PubMed

    Yin, Guangwen; Lin, Qian; Qiu, Jianhan; Qin, Mei; Tang, Xinming; Suo, Xun; Huang, Zhijian; Liu, Xianyong

    2015-05-30

    The CD40 ligand (CD40L) has shown potential as a powerful immunological adjuvant in various studies. Here, the efficacy of a chimeric subunit vaccine, consisting of Eimeria tenella immune mapped protein 1 (EtIMP1) and chicken CD40L, was evaluated against E. tenella infection. The recombinant EtIMP1-CD40L was purified from E. coli over-expressing this protein. Chickens were vaccinated with EtIMP1-CD40L without adjuvant or EtIMP1 with Freund's adjuvant. Immunization of chickens with EtIMP1-CD40L fusion protein resulted in stronger IFN-γ secretion and IgA response than that with only recombinant EtIMP1 with Freund's adjuvant. The clinical effect (cecal lesions, body weights gain, and oocysts shedding) of the EtIMP1-CD40L without adjuvant was also better than that of the EtIMP1 with adjuvant, as evidenced by the difference between the two groups in the oocyst output of E. tenella-challenged chickens. The results suggest that the EtIMP1-CD40L fusion protein can be used as an effective immunogen in the development of subunit vaccines against Eimeria infection.

  1. Characterization of a developmentally regulated oocyst protein from Eimeria tenella.

    PubMed

    Fetterer, R H; Barfield, R C

    2003-06-01

    Changes in proteins during sporulation of Eimeria tenella oocysts were investigated. Unsporulated E. tenella oocysts collected from cecal tissue at 7 days postinoculation were sporulated in aerated media at 28 C for 0-48 hr. Gel analysis of soluble protein extracts prepared from oocysts from their respective time points indicated the presence of 2 prominent bands with relative molecular weight (Mr) in the range of 30 kDa and making up 20% of the total protein. These 2 bands, designated as major oocyst proteins (MOPs), were absent or barely detectable by 21 hr of sporulation. MOP bands were weakly reactive with glycoprotein stain but showed no mobility shift on deglycosylation. By gel analysis it was shown that the purified MOPs consisted of 2 bands of Mr 28.7 and 30.1 kDa. However, by matrix-assisted laser deabsorption-time of flight analysis it was shown that masses were about 17% lower. Internal sequence analysis of the 28.7-kDa protein generated 2 peptides of 17 and 14 amino acids in length, consistent with a recently described protein coded by the gam56 gene and expressed in E. maxima gametocytes. Rabbit antibodies made against MOPs were localized to outer portions of sporocysts before excystment and to the apical end of in vitro-derived sporozoites. These same antibodies were found to react with bands of Mr 101 and 65 kDa by Western blot but did not recognize MOPs in soluble or insoluble sporozoite extracts. The data suggest that the MOPs are derived from part of a gametocyte protein similar to that coded by gam56 and are processed during sporulation into sporocyst and sporozoite proteins. Alternatively, the binding of anti-MOP to 101- and 65-kDa proteins may result from alternatively spliced genes as the development of parasite proceeds.

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

    PubMed

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

    2013-10-25

    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.

  3. Immune responses to dietary beta-glucan in broiler chicks during an Eimeria challenge.

    PubMed

    Cox, C M; Sumners, L H; Kim, S; McElroy, A P; Bedford, M R; Dalloul, R A

    2010-12-01

    Escalating consumer concerns regarding pathogen resistance have placed the poultry industry under mounting pressure to eliminate the use of chemotherapeutic agents as feed additives. One possible alternative receiving increased attention is the use of immunomodulators such as β-glucan. A study was conducted to investigate the effects of a yeast-derived β-glucan (Auxoferm YGT) on broiler chick performance, lesion scores, and immune-related gene expression during a mixed Eimeria infection. Day-old chicks were fed diets containing 0, 0.02, or 0.1% YGT. On d 8 posthatch, one-half of the replicate pens were challenged with a mixed inoculum of Eimeria acervulina, Eimeria maxima, and Eimeria tenella. Measurements were taken and samples collected on d 4, 10, 14, and 21 posthatch. Dietary supplementation had no effect on performance or mortality. On d 14, 3 birds per pen (n = 24/treatment) were scored for intestinal coccidia lesions. Gross lesion severity was significantly reduced in birds supplemented with 0.1% YGT. On d 10, inducible nitric oxide synthase (iNOS) expression was downregulated in the jejunum of challenged birds fed 0.1% YGT. Expression of iNOS in the ileum was downregulated in the nonchallenged birds, but upregulated in the challenged birds fed 0.1% YGT on d 14. Interleukin (IL)-18 was upregulated in the jejunum of 0.1% YGT-treated birds. Interferon (IFN)-γ expression was decreased in challenged and nonchallenged birds fed 0.1% YGT. The IL-4 expression was downregulated in the nonchallenged birds with 0.1% YGT diet supplementation. The IL-13 and mucin-1 levels were also reduced due to β-glucan supplementation. Mucin-2 expression was increased in the nonchallenged birds, but decreased in the infected birds fed 0.1% YGT. These results suggest that although Auxoferm YGT at doses of 0.02 and 0.1% does not influence performance, it significantly reduces lesion severity and is capable of altering immune-related gene expression profiles, favoring an enhanced T

  4. Persistence of Eimeria bovis in soil.

    PubMed

    Lassen, Brian; Lepik, Triin; Bangoura, Berit

    2013-07-01

    Eimeriosis is a disease that occurs globally and often affects cattle grazing on pastures contaminated with oocysts of the pathogenic species Eimeria bovis, Eimeria zuernii or Eimeria alabamensis, respectively. Nonetheless, little is understood regarding oocyst persistence on the pasture. The study was performed in the temperate climate zone. Soil samples were spiked with 100,000 E. bovis oocysts in July 2010 or with 50,000 oocysts in October 2010, respectively, both either with our without addition of cattle faeces. The soil samples were exposed to natural environmental conditions until April 2011. A subset of the samples was analysed immediately after spiking as positive control. The oocysts were recovered by a flotation method and counted in a reading chamber. On average, 23 % of the oocysts could be recovered from the positive control. The recovery of oocysts dropped to 0.30 % of the original level in the samples prepared in July independent of the addition of faeces, whereas the oocyst count was higher in the samples prepared in October, both without (2.05 %) and with (2.64 %) faecal material. No differences were observed between presence of oocysts or oocyst counts recovered in the presence or absence of faeces. Presence of faeces had a positive influence on oocyst integrity. During the winter season, the number of oocysts in the soil was lowered under the detection limit in most samples. On the other hand, the comparatively short 3-month summer period had a significant influence on the number of persisting oocysts too.

  5. [Eimeria of cattle in the Republic of Dagestan].

    PubMed

    Usarova, E I

    2008-01-01

    Data on Eimeria (Sporozoa: Coccidia) parasitizing cattle in different regions of the Republic of Dagestan are given. Eight species of the genus were found. Eimeria zuernii, E. subspherica, and E. bovis appeared to be the dominate species. Total infestation rate in the republic is 67.7%, while in mountain regions of Dagestan in is 83%.

  6. In vitro destruction of Eimeria oocysts by essential oils.

    PubMed

    Remmal, Adnane; Achahbar, Sanaa; Bouddine, Latifa; Chami, Najat; Chami, Fouzia

    2011-12-15

    This study aims to assess the ability of essential oils (EOs) to destroy Eimeria oocyst in vitro using microscopic counting and 273 nm absorbing material release. A screening for the ability of ten EOs to destroy Eimeria oocyst was carried out in liquid medium. Out of these ten, artemisia, tea tree, thyme and clove EOs were identified as being the most effective. The treatment of Eimeria oocyst with these EOs leads to their lysis as shown by the release of substances absorbing at 273 nm. These results were obtained after approximately three hours contact. Four EOs were proven to destroy Eimeria oocysts in a few hours at low concentration. This destructive effect is a consequence of their lysis. This work is a preliminary contribution aiming to develop a new generation of natural efficient agents for destroying Eimeria oocyst to fight coccidiosis in broiler chicken. Copyright © 2011 Elsevier B.V. All rights reserved.

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

    PubMed

    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.

  8. Growth performance and gastrointestinal responses of broiler chickens fed corn-soybean meal diet without or with exogenous epidermal growth factor upon challenge with Eimeria.

    PubMed

    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 < 0.05) increased BWG before challenge. There was no EGF and Eimeria interaction (P > 0.05) on growth performance, AR of GE, and intestinal histomorphology; the main effects were such that Eimeria depressed (P < 0.01) BWG, FCR, AR of DM, crude fat, and GE, and villi height to crypt depth ratio. An interaction between EGF and Eimeria (P < 0.05) on indices of gut function was such that EGF improved expression of genes for nutrient transporters and tight junction proteins in 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

  9. Molecular cloning and functional expression of mannitol-1-phosphatase from the apicomplexan parasite Eimeria tenella.

    PubMed

    Liberator, P; Anderson, J; Feiglin, M; Sardana, M; Griffin, P; Schmatz, D; Myers, R W

    1998-02-13

    A metabolic pathway responsible for the biosynthesis and utilization of mannitol is present in the seven species of Eimeria that infect chickens, but is not in the avian host. Mannitol-1-phosphatase (M1Pase), a key enzyme for mannitol biosynthesis, is a highly substrate-specific phosphatase and, accordingly, represents an attractive chemotherapeutic target. Amino acid sequence of tryptic peptides obtained from biochemically purified Eimeria tenella M1Pase was used to synthesize degenerate oligonucleotide hybridization probes. Using these reagents, a partial genomic clone and full-length cDNA clones have been isolated and characterized. The deduced amino acid sequence of E. tenella M1Pase shows limited overall homology to members of the phosphohistidine family of phosphatases. This limited homology to other histidine phosphatases does, however, include several conserved residues that have been shown to be essential for their catalytic activity. Kinetic parameters of recombinant M1Pase expressed in bacteria are essentially identical to those of the biochemically purified preparation from E. tenella. Moreover, recombinant M1Pase is subject to active site-directed, hydroxylamine-reversible inhibition by the histidine-selective acylating reagent diethyl pyrocarbonate. These results indicate the presence of an essential histidine residue(s) at the M1Pase active site, as predicted for a histidine phosphatase.

  10. Effect of amprolium on production, sporulation, and infectivity of Eimeria oocysts.

    PubMed

    Ruff, M D; Garcia, R; Chute, M B; Tamas, T

    1993-01-01

    Amprolium reduced the number of oocysts shed by Eimeria acervulina, E. maxima, E. necatrix, and a mixture of susceptible strains of E. tenella. Sporulation of oocysts from mediated chickens was reduced compared with that of oocysts from unmedicated chickens. Sporulation was reduced by levels of 0.0250% amprolium for E. acervulina and by levels of 0.0060% for E. maxima and the susceptible E. tenella. Not enough oocysts were recovered to measure sporulation of E. necatrix. Sporulation reduction was not affected by the method of administration of amprolium (feed or water), except with E. acervulina, for which fewer oocysts sporulated when 0.0120% amprolium was added in the drinking water than when 0.0125% amprolium was added to the feed. Conversely, amprolium medication had no effect on the sporulation of an amprolium-resistant E. tenella. When fed to unmedicated chickens, those oocysts from amprolium-medicated chickens that did sporulate were as infective as oocysts recovered from unmedicated chickens.

  11. Anticoccidial efficacy of diclazuril against recent field isolates of Eimeria from commercial poultry farms.

    PubMed

    McDougald, L R; Mathis, G F; Seibert, B P

    1990-01-01

    The anticoccidial efficacy of diclazuril, a novel anticoccidial agent, was titrated in laboratory experiments using recent field isolates of Eimeria. Fifty tests were conducted with six individual species isolates, and seven tests were done with a mixture of the six species. Results were based on intestinal lesion scores at necropsy, droppings scores, and weight gain. Diclazuril at 0.5 ppm was almost completely effective against E. tenella, E. acervulina, and E. mitis. Prevention of E. brunetti was better at 1.0 ppm than at 0.5 ppm. In birds infected with E. mitis. Prevention of E. brunetti was better at 1.0 ppm than at 0.5 ppm. In birds infected with E. maxima, diclazuril at 0.5-1.5 ppm significantly reduced lesion scores and droppings scores and improved weight gain, although lesions were higher than with other species. Oocyst shedding by E. maxima was almost completely prevented by 0.5-1.5 ppm. Lesion scores and droppings scores caused by E. necatrix or mixed infections were greatly reduced by 0.5 ppm of diclazuril, but 1.0 ppm was necessary to obtain full protection of weight gain. Results suggest that 1.0 ppm of diclazuril best prevents coccidiosis caused by six species of coccidia in chickens.

  12. Immune response of broiler chickens fed different levels of arginine and vitamin E to a coccidiosis vaccine and Eimeria challenge.

    PubMed

    Perez-Carbajal, C; Caldwell, D; Farnell, M; Stringfellow, K; Pohl, S; Casco, G; Pro-Martinez, A; Ruiz-Feria, C A

    2010-09-01

    One-day-old broiler chicks (n = 300) were orally vaccinated (Coccivac-B) and divided into 6 groups to evaluate Arg at 3 levels of supplementation, 0, 0.3, or 0.6% [normal level (NARG), medium level (MARG), or high level (HARG), respectively], and 2 levels of vitamin E (VE), 40 or 80 IU/kg of feed (VE40 or VE80, respectively), in a factorial experiment. Birds were reared in floor pens with fresh pine shavings and provided a corn-soybean-based diet and water ad libitum. At d 14, all chickens were orally challenged with a mixture of Eimeria field isolates (Eimeria acervulina, Eimeria maxima, and Eimeria tenella). In vitro heterophil and monocyte oxidative burst (HOB and MOB, respectively) was measured at d 21 from cells isolated from peripheral blood. Antibody levels (IgG, IgM, and IgA isotypes, ELISA) and NO were measured at d 14 and 28. The HOB was lower in birds fed the VE40 diets but was increased with the MARG and HARG treatments, whereas birds fed the VE80 diet had a higher HOB irrespective of Arg level. Birds fed the VE80 diet had high levels of MOB, which was not further improved by Arg, whereas birds fed the VE40-MARG diet had the highest MOB response. Plasma NO was not affected by diet at d 14, but at d 28, plasma NO was higher in birds fed the VE80-MARG or the VE40-NARG diet and lower in birds fed the VE80-NARG or the VE40-MARG diet. Birds fed the VE40-HARG or VE80-MARG diet had the highest IgG levels at d 14, but at d 28, birds fed the VE80-MARG diet had the highest IgG levels. The IgM concentration was lower in birds fed NARG levels irrespective of VE levels at d 14, but at d 28, IgM levels were higher in birds fed the VE40-HARG or the VE80-MARG feed. The IgA concentration was not consistently affected at d 14 or 28. These results suggest that Arg and VE fed at levels higher than those recommended by the NRC may play complementary roles on the innate and humoral immune response against an Eimeria challenge, potentially improving vaccine efficacy and

  13. Description of Eimeria pavonina (coccidia) of peafowl in Germany.

    PubMed

    Hauck, Rüdiger; Hafez, Hafez M

    2012-03-01

    There are only a few reports about the occurrence of coccidia in peafowl and no reports about the occurrence of Eimeria spp. in peafowl kept in Europe. Here, we describe the occurrence of Eimeria pavonina in diseased peafowl from Germany. In January 2011, one young peacock kept in an aviary showed a marked depression. No parasites were detected in samples from the diseased bird, but in samples of birds from the same and other aviaries, coccidian counts were between 400/g and 66,000/g. All peacocks were treated with toltrazuril. After treatment, the clinical condition of the diseased bird improved but, two weeks afterwards, other birds in the aviary were still shedding coccidia in their feces. Based on morphology, the coccidia were identified as E. pavonina. Parts of the 18s rRNA gene and the cytochrome oxidase subunit 1 (cox-1) gene were sequenced. A phylogenetic tree based on the 18s rRNA sequence placed the Eimeria sp. from peafowl closest to Eimeria spp. found in pheasants and partridges as well as to Eimeria meleagrimitis. A phylogenetic tree based on the sequence of cox-1 in contrast suggested a closer relationship to Eimeria necatrix and Eimeria tenella.

  14. Epidemiology of Eimeria and associated risk factors in cattle of district Toba Tek Singh, Pakistan.

    PubMed

    Rehman, Tauseef Ur; Khan, Muhammad Nisar; Sajid, Muhammad Sohail; Abbas, Rao Zahid; Arshad, Muhammad; Iqbal, Zafar; Iqbal, Asif

    2011-05-01

    A cross-sectional study was carried out to determine the prevalence, species characterization, and associated risk factors with Eimeria of cattle of district Toba Tek Singh from April, 2009 to March, 2010. Of the total 584 fecal samples examined for Eimeria, 275 (47.09%) were found infected with six species of Eimeria. Among the identified species of Eimeria, Eimeria bovis was found to be the highest prevalent species (52.36%), followed in order by Eimeria zuernii, Eimeria canadensis, Eimeria ellipsoidalis, Eimeria alabamensis, and Eimeria cylindrica with prevalence of 48.27%, 34.83%, 29.31%, 24.14%, and 8.62% respectively. Peak prevalence was observed in August. Cattle were infected more frequently during rainy (60.32%) and post-rainy seasons (59.25%). Calves had significantly higher prevalence (P<0.05) of Eimeria than adults while higher prevalence of Eimeria was observed in female cattle. Among management and husbandry practices, feeding system, watering system, housing system, floor type, and herd size strongly influenced the prevalence of Eimeria in cattle. Coccidiosis was more prevalent in ground feeding system, pond-watered animals, closed housing system, and non-cemented floor type (P<0.05) as compared to trough feeding system, tap watered animals, open housing system, and partially cemented floor types, respectively. Breed and body condition of animals were not found risk factors (P>0.05) influencing prevalence of Eimeria.

  15. Eimeria oocyst concentrations and species composition in litter from commercial broiler farms during anticoccidial drug or live Eimeria oocyst vaccine control programs

    USDA-ARS?s Scientific Manuscript database

    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 ooc...

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

    PubMed

    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.

  17. Multiple thermal maxima during the Holocene

    SciTech Connect

    Davis, O.K.

    1984-08-10

    The astronomical theory of climatic change provides an alternative to the traditional chronology for Holocene climatic change, which calls for one thermal maximum about 6000 years ago. The theory predicts a series of maxima during the Holocene, one for each season. Because the relation of the perihelion to the spring equinox changes with a 22,000-year period, late summer insolation would have been greatest 5000 years ago, whereas early summer insolation would have been greatest 13,000 years ago. Climatic reconstructions based on the response of ecosystems to late summer climate indicate a later Holocene thermal maximum than paleoclimatic data sensitive to early summer climate. In southern Idaho, three different vegetation types indicate thermal maxima at different times during the Holocene, depending on the climatic variable controlling each type. 28 references, 2 figures.

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

    PubMed

    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.

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

    PubMed Central

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

    2015-01-01

    Aim: To determine the prevalence and diversity of Eimeria spp. in dairy cattle present in and around Guwahati, Kamrup district, Assam, India. Materials and Methods: A total of 2339 fecal samples of calves (535), heifer (641) and adult (1163) cattle were screened for 1 year present in and around Guwahati, Assam for detection of Eimeria oocysts by flotation techniques. Sporulation of the oocyst was done in 2.5% potassium dichromate solution for identification of the Eimeria species. Results: 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 (<1 year), heifer (1-3 years) and adult (>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. PMID:27047181

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

    PubMed

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

    2014-07-17

    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 mitotically replicating mitochondrial genome may be more suited for species level molecular work; however, only limited sequence data are available for Eimeria spp. infecting turkeys. The purpose of this study was to sequence and annotate the complete mitochondrial genomes from 5 Eimeria species that commonly infect the domestic turkey (Meleagris gallopavo). Six single-oocyst derived cultures of five Eimeria species infecting turkeys were PCR-amplified and sequenced completely prior to detailed annotation. Resulting sequences were aligned and used in phylogenetic analyses (BI, ML, and MP) that included complete mitochondrial genomes from 16 Eimeria species or concatenated CDS sequences from each genome. Complete mitochondrial genome sequences were obtained for Eimeria adenoeides Guelph, 6211 bp; Eimeria dispersa Briston, 6238 bp; Eimeria meleagridis USAR97-01, 6212 bp; Eimeria meleagrimitis USMN08-01, 6165 bp; Eimeria gallopavonis Weybridge, 6215 bp; and Eimeria gallopavonis USKS06-01, 6215 bp). The order, orientation and CDS lengths of the three protein coding genes (COI, COIII and CytB) as well as rDNA fragments encoding ribosomal large and small subunit rRNA were conserved among all sequences. Pairwise sequence identities between species ranged from 88.1% to 98.2%; sequence variability was concentrated within CDS or between rDNA fragments (where indels were common). No phylogenetic reconstruction supported monophyly of Eimeria species infecting turkeys; Eimeria dispersa may have arisen

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

    PubMed Central

    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 mitotically replicating mitochondrial genome may be more suited for species level molecular work; however, only limited sequence data are available for Eimeria spp. infecting turkeys. The purpose of this study was to sequence and annotate the complete mitochondrial genomes from 5 Eimeria species that commonly infect the domestic turkey (Meleagris gallopavo). Methods Six single-oocyst derived cultures of five Eimeria species infecting turkeys were PCR-amplified and sequenced completely prior to detailed annotation. Resulting sequences were aligned and used in phylogenetic analyses (BI, ML, and MP) that included complete mitochondrial genomes from 16 Eimeria species or concatenated CDS sequences from each genome. Results Complete mitochondrial genome sequences were obtained for Eimeria adenoeides Guelph, 6211 bp; Eimeria dispersa Briston, 6238 bp; Eimeria meleagridis USAR97-01, 6212 bp; Eimeria meleagrimitis USMN08-01, 6165 bp; Eimeria gallopavonis Weybridge, 6215 bp; and Eimeria gallopavonis USKS06-01, 6215 bp). The order, orientation and CDS lengths of the three protein coding genes (COI, COIII and CytB) as well as rDNA fragments encoding ribosomal large and small subunit rRNA were conserved among all sequences. Pairwise sequence identities between species ranged from 88.1% to 98.2%; sequence variability was concentrated within CDS or between rDNA fragments (where indels were common). No phylogenetic reconstruction supported monophyly of Eimeria species infecting turkeys

  2. Identification and immunogenicity of microneme protein 2 (EbMIC2) of Eimeria brunetti.

    PubMed

    Hoan, Tran Duc; Zhang, Zhenchao; Huang, Jingwei; Yan, Ruofeng; Song, Xiaokai; Xu, Lixin; Li, Xiangrui

    2016-03-01

    There have been only a few antigen genes of Eimeria brunetti reported up to now. In this study, the gene encoding the microneme protein 2 (EbMIC2) was isolated from oocysts of E. brunetti by RT-PCR and the immunogenicity of recombinant EbMIC2 was observed. The EbMIC2 was cloned into vector pMD19-T for sequencing. The sequence was compared with the published EbMIC2 gene from GenBank revealed homology of the nucleotide sequence and amino acids sequence were 99.43 and 98.63%, respectively. The correct recombinant pMD-EbMIC2 plasmid was inserted into the pET-28a (+) expressing vector and transformed into competent Escherichia coli BL21 cells for expression. The expressed product was analyzed using SDS-PAGE and Western-blot. The results indicated that the recombinant EbMIC2 protein was recognized strongly by serum from naturally infected chicken with E. brunetti. Rat rcEbMIC2 antisera bound to bands of about 36 kDa in the somatic extract of E. brunetti sporozoites. The recombinant plasmid pVAX1-EbMIC2 was constructed and then the efficacies of recombinant plasmid and recombinant protein were evaluated. The results of IgG antibody level and cytokines concentration suggested that recombinant EbMIC2 could increase the IgG antibody level and induce the expressions of cytokines. Animal challenge experiments demonstrated that the recombinant EbMIC2 protein and recombinant plasmid pVAX1-EbMIC2 could significantly increase the average body weight gains, decrease the mean lesion scores and the oocyst outputs of the immunized chickens and presented high anti-coccidial index. All results suggested that EbMIC2 could become an effective candidate for the development of new vaccine against E. brunetti infection.

  3. A simple method for the purification of Eimeria tenella sporozoites.

    PubMed

    Mattig, F R; Drössigk, U; Entzeroth, R

    1993-05-01

    A rapid, simple and cheap method for the purification of Eimeria tenella sporozoites has been developed using commercially available filter paper (595 filter paper circles, Schleicher & Schuell, 3354 Dassel, Germany, order 311610). Yield and purity are of the same value as by purification with a leukopac column (Bontemps & Yvore 1974). The described method can be used for the purification of Eimeria tenella sporozoites under sterile conditions for subsequent in vitro cultivation.

  4. Prevalence of Eimeria bovis and Eimeria zuernii in German cattle herds and factors influencing oocyst excretion.

    PubMed

    Bangoura, Berit; Mundt, Hans-Christian; Schmäschke, Ronald; Westphal, Bernhard; Daugschies, Arwid

    2012-02-01

    The present study was designed to investigate the prevalence of the pathogenic coccidia species Eimeria bovis and Eimeria zuernii in shed-reared animals in German dairy and fattening facilities. Samples were obtained from 65 cattle farms distributed randomly across all the regions of Germany regardless of the occurrence of clinical problems. The samples were obtained rectally. Faecal consistency and the total number of oocysts per gram of faeces (OPG) were determined, along with the OPG values for E. bovis and E. zuernii. A questionnaire was completed for each farm to record information about herd size and management, along with individual animal data. Eimeria oocysts were detected in 62 of these farms, which give a prevalence of 95.4%. The farm prevalence of the pathogenic species was 76.9% for E. bovis and 83.1% for E. zuernii. The number of oocysts excreted could not be correlated significantly with farm type or farm management but depended on the floor type, the age of the calves and the time after rehousing. Furthermore, there was a positive correlation between OPG and the observation of diarrhoea. E. zuernii had a greater influence on the occurrence of diarrhoea than E. bovis. This study confirms that herd management frequently does not meet the requirements of effective coccidia control despite the fact that the pathogenic coccidia E. bovis and E. zuernii are ubiquitous in German cattle populations.

  5. Effects of feed additives and mixed eimeria species infection on intestinal microbial ecology of broilers.

    PubMed

    Hume, M E; Clemente-Hernández, S; Oviedo-Rondón, E O

    2006-12-01

    Evaluation of digestive microbial ecology is necessary to understand effects of growth-promoting feed. In the current study, the dynamics of intestinal microbial communities (MC) were examined in broilers fed diets supplemented with a combination of antibiotic (bacitracin methylene disalicylate) and ionophore (Coban 60), and diets containing 1 of 2 essential oil (EO) blends, Crina Poultry (CP) and Crina Alternate (CA). Five treatments were analyzed: 1) unmedicated uninfected control; 2) unmedicated infected control; 3) feed additives monensin (bacitracin methylene disalicylate) + monensin (Coban 60; AI); 4) EO blend CP; and 5) EO blend CA. Additives were mixed into a basal feed mixture, and EO were adjusted to 100 ppm. Chicks were infected by oral gavage at 19 d of age with Eimeria acervulina, Eimeria maxima, and Eimeria tenella. Duodenal, ileal, and cecal samples were taken from 12 birds per treatment just before and 7 d after challenge; 2 samples each were pooled to give a final number of 6 samples total; and all pooled samples were frozen until used for DNA extraction. Denaturing gradient gel electrophoresis was used to examine PCR-amplified fragments of the bacterial 16S ribosomal DNA variable region. Results are presented as percentages of similarity coefficients (SC). Dendrograms of PCR amplicon or band patterns indicated MC differences due to intestinal location, feed additives, and cocci challenge. Essential oil blends CP and CA affected MC in all gut sections. Each EO had different effects over MC, and they differed in most instances from the AI group. The cocci challenge caused drastic MC population shifts in duodenal, ileal, and cecal sections (36.7, 55.4, and 36.2% SC, respectively). Diets supplemented with CP supported higher SC between pre- and postchallenge MC (89.9, 83.3, and 76.4%) than AI (81.8., 57.4, and 60.0%). We concluded that mixed coccidia challenge caused drastic shifts in MC. These EO blends modulated MC better than AI, avoiding drastic

  6. Eimeria infections in litter-based, high stocking density systems for loose-housed laying hens in Sweden.

    PubMed

    Lundén, A; Thebo, P; Gunnarsson, S; Hooshmian-Rad, P; Tauson, R; Uggla, A

    2000-09-01

    1. Coccidiosis, caused by different Eimeria species, is believed to be a more prominent problem in loose-housed layers kept on litter than in battery cages. In this study, the impact and development of Eimeria infections were investigated in layers kept in litter-based, high stocking density systems for loose-housed hens. 2. Layers from 57 flocks on 26 farms were followed by necropsy of a representative sample of birds that died or had to be culled. Coccidiosis was diagnosed in 11 flocks (19.3%) from 9 (31%) of the farms. The outbreaks occurred when the birds were 19 to 32 weeks old. E. maxima was identified in 6 and E. tenella in 3 of the outbreaks. 3. Sixteen of the flocks were also monitored with faecal and litter samples collected at regular intervals. Oocysts were detected in samples from all these flocks. The pattern of oocyst excretion was similar in most of the flocks, with maximum counts at 4 to 8 weeks after introduction to the laying house. There was no significant correlation between the levels of oocysts in faeces and clinical coccidiosis. 4. Raising pullets without any coccidiostat, to increase their chance to develop immunity against coccidia, was not found to decrease the risk of coccidiosis during the production period when compared to the practice of giving amprolium and ethopabate during the rearing period.

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

    USDA-ARS?s Scientific Manuscript database

    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...

  8. Type I and type II fatty acid biosynthesis in Eimeria tenella: enoyl reductase activity and structure.

    PubMed

    Lu, J Z; Muench, S P; Allary, M; Campbell, S; Roberts, C W; Mui, E; McLeod, R L; Rice, D W; Prigge, S T

    2007-12-01

    Apicomplexan parasites of the genus Eimeria are the major causative agent of avian coccidiosis, leading to high economic losses in the poultry industry. Recent results show that Eimeria tenella harbours an apicoplast organelle, and that a key biosynthetic enzyme, enoyl reductase, is located in this organelle. In related parasites, enoyl reductase is one component of a type II fatty acid synthase (FAS) and has proven to be an attractive target for antimicrobial compounds. We cloned and expressed the mature form of E. tenella enoyl reductase (EtENR) for biochemical and structural studies. Recombinant EtENR exhibits NADH-dependent enoyl reductase activity and is inhibited by triclosan with an IC50 value of 60 nm. The crystal structure of EtENR reveals overall similarity with other ENR enzymes; however, the active site of EtENR is unoccupied, a state rarely observed in other ENR structures. Furthermore, the position of the central beta-sheet appears to block NADH binding and would require significant movement to allow NADH binding, a feature not previously seen in the ENR family. We analysed the E. tenella genomic database for orthologues of well-characterized bacterial and apicomplexan FAS enzymes and identified 6 additional genes, suggesting that E. tenella contains a type II FAS capable of synthesizing saturated, but not unsaturated, fatty acids. Interestingly, we also identified sequences that appear to encode multifunctional type I FAS enzymes, a feature also observed in Toxoplasma gondii, highlighting the similarity between these apicomplexan parasites.

  9. Prevalence of Eimeria bovis and Eimeria zuernii in German cattle herds and factors influencing oocyst excretion.

    PubMed

    Bangoura, Berit; Mundt, Hans-Christian; Schmäschke, Ronald; Westphal, Bernhard; Daugschies, Arwid

    2011-08-01

    The present study was designed to investigate the prevalence of the pathogenic coccidia species E. bovis and E. zuernii in shed-reared animals in German dairy and fattening facilities.Samples were obtained from 65 cattle farms distributed randomly across all the regions of Germany, regardless of the occurrence of clinical problems. The samples were obtained rectally. Faecal consistency and the total number of oocysts per gram of faeces (OPG) were determined for Eimeria spp., along with the separate OPG values for Eimeria (E.) bovis and E. zuernii. A questionnaire was completed for each farm to record information about herd size and management together with individual animal data. Eimeria oocysts, regardless of the kind of Eimeria spp., were detected in 62 of these farms, which gives a prevalence of 95.4 %. The farm prevalence of the pathogenic species was 76.9 % for E. bovis and 83.1 % for E. zuernii. The average oocyst excretion level was 2,950 OPG in terms of total Eimeria spp. oocyst excretion, 700 OPG for E. bovis and 1,500 OPG for E. zuernii.The number of oocysts excreted could not be correlated significantly with farm type or farm management but depended on the floor type which influences the infection pressure, on the age of the calves and the time after rehousing. In general, higher oocyst excretion rates were found in calves kept on litter compared to rearing on slatted floor. Younger calves and calves sampled early after housing shed higher amounts of oocysts than older calves and calves stabled a longer period before sampling, respectively. Furthermore, there was a positive correlation between OPG and the observation of diarrhoea, defined as observation of a loose to liquid faecal consistency. Excretion of E. zuernii oocysts was more closely linked to the occurrence of diarrhoea than E. bovis oocyst excretion. This study confirms that the pathogenic coccidia E. bovis and E. zuernii are ubiquitous in German cattle populations and a significant cause of

  10. Suppression of Eimeria tenella sporulation by disinfectants.

    PubMed

    You, Myung-Jo

    2014-08-01

    The disinfectant effects (DEs) of 10 types of chemicals, defined by their ability to destroy or inhibit oocysts and consequently prevent sporulation of Eimeria tenella field isolate, were evaluated in vitro. Correct species assignments and sample purities were confirmed by the singular internal transcribed spacer (ITS)-PCR analysis. A total of 18 treatments were performed, and the disinfection suppression levels were 75.9% for 39% benzene + 22% xylene (1:10 dilution), 85.5% for 30% cresol soup (1:1 dilution), and 91.7% for 99.9% acetic acid (1:2 dilution) group. The results indicate that acetic acid, cresol soup, and benzene+xylene are good candidates for suppression of E. tenella oocyst sporulation.

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

    PubMed

    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. Vaccination with Clostridium perfringens recombinant proteins in combination with Montanide™ ISA 71 VG adjuvant increases protection against experimental necrotic enteritis in commercial broiler chickens.

    PubMed

    Jang, Seung I; Lillehoj, Hyun S; Lee, Sung-Hyen; Lee, Kyung Woo; Lillehoj, Erik P; Hong, Yeong Ho; An, Dong-Jun; Jeong, Wooseog; Chun, Ji-Eun; Bertrand, François; Dupuis, Laurent; Deville, Sébastien; Arous, Juliette Ben

    2012-08-03

    This study was performed to compare four Clostridium perfringens recombinant proteins as vaccine candidates using the Montanide™ ISA 71 VG adjuvant in an experimental model of necrotic enteritis. Broiler chickens were immunized subcutaneously with purified clostridial recombinant NetB toxin, pyruvate: ferredoxin oxidoreductase (PFO), α-toxin, or elongation factor-Tu (EF-Tu), or with vehicle control, in conjunction with ISA 71 VG, and intestinal lesion scores, body weight gains, NetB toxin and PFO antibody levels, and proinflammatory cytokine and chemokine levels were measured as outcomes of protection following oral co-infection with C. perfringens and Eimeria maxima. Birds immunized with all recombinant proteins plus ISA 71 VG showed significantly reduced gut lesions compared with the ISA 71 VG-only group. Birds immunized with NetB toxin or PFO plus ISA 71 VG exhibited significantly increased body weight gains compared with the ISA 71 VG alone group. Greater NetB toxin antibody titers were observed in the NetB/ISA 71 VG group, and greater PFO antibody titers were evident in the PFO/ISA 71 VG group, each compared with the other three vaccine/adjuvant groups. Finally, decreased levels of gene transcripts encoding interleukin-8, tumor necrosis factor superfamily 15, and LPS-induced TNF-α factor were observed in the intestinal lymphocytes of chickens immunized with NetB toxin, PFO, α-toxin, and/or EF-Tu in the presence of ISA 71 VG compared with ISA 71 VG alone. All parameters evaluated were equal in co-infected chickens given ISA 71 VG alone compared with infected/adjuvant-free birds, indicating that the adjuvant itself did not have a disease protective effect. These results suggest that vaccination with clostridial recombinant proteins, particularly NetB toxin or PFO, in combination with ISA 71 VG enhances protective immunity against experimental necrotic enteritis in broiler chickens. Published by Elsevier Ltd.

  13. Chimeric DNA vaccines encoding Eimeria acervulina macrophage migration inhibitory factor (E.MIF) induce partial protection against experimental Eimeria infection.

    PubMed

    Song, Xiaokai; Zhang, Ruirui; Xu, Lixin; Yan, Ruofeng; Li, Xiangrui

    2015-09-01

    Chimeric DNA vaccines co-expressing Eimeria acervulina macrophage migration inhibitory factor (E.MIF) and chicken IL-2 (IL-2) or interferon-γ (IFN-γ) were constructed and their efficacies against E. acervulina were evaluated. The open reading frame (ORF) of E.MIF was cloned from E. acervulina merozoites and subcloned into the eukaryotic expression vector pVAX1 with chicken cytokine gene IFN-γ or IL-2 to construct the DNA vaccines pVAX-E.MIF-IFN-γ, pVAX-E.MIF-IL-2 and pVAX-E.MIF. The in vivo transfection of the target genes was detected by use of reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Immunizations were carried out by vaccinating chickens twice with a dose rate of 100 μg intramuscularly. Seven days post second immunization, all chickens except the unchallenged control group were challenged orally with 1 × 105 sporulated oocysts of E. acervulina. Seven days later, the duodenum was collected. The results showed that the target genes were expressed effectively in vivo. DNA vaccines and the recombinant E.MIF protein could alleviate body weight loss and duodenal lesions significantly compared to the control groups. Furthermore, pVAX-E.MIF-IL-2 and pVAX-E.MIF-IFN-γ induced anticoccidial indexs (ACIs) of 179.12 and 170, respectively, which were significantly higher than that of pVAX-E.MIF (ACI = 162.31). Our results demonstrated that E.MIF is a potential vaccine candidate against E. acervulina and chicken IFN-γ or IL- 2 may be used as genetic adjuvants to improve the efficacies of DNA vaccines against avian coccidiosis.

  14. Intestinal microbial ecology of broilers vaccinated and challenged with mixed Eimeria species, and supplemented with essential oil blends.

    PubMed

    Oviedo-Rondón, E O; Hume, M E; Hernández, C; Clemente-Hernández, S

    2006-05-01

    Intestinal microbiota is an important component in the development of defense mechanisms in the gut mucosa. This project determined the dynamics of intestinal microbial communities (MC) of broilers vaccinated at first day of age with live oocysts of Eimeria species and fed diets supplemented with 2 specific essential oil (EO) blends, Crina Poultry (CP) and Crina Alternate (CA). Five treatments were analyzed: 1) unmedicated-uninfected (UU) control; 2) unmedicated-infected (UI) control; 3) vaccinated with Advent cocci-vaccine and without feed additive (COV) supplements; 4) vaccinated with Advent and supplemented with CP; and 5) vaccinated with Advent and supplemented with CA. The EO blends were added at 100 ppm to the same basal diets. Chicks were gavage-infected at 19 d of age with Eimeria acervulina, Eimeria maxima, and Eimeria tenella. Duodenal, ileal, and cecal samples were taken from 12 birds per treatment just before the infection and 7 d after the challenge, pooled in 6 samples, and frozen. Denaturing gradient gel electrophoresis was used to examine PCR-amplified fragments of the bacterial 16S ribosomal DNA variable region. Results are presented as percentages of similarity coefficients (SC). Dendrograms of amplicon patterns indicated MC differences due to intestinal location, feed additives, and cocci infection. The EO blends CP and CA did affect MC in all gut sections. The cocci-infection caused drastic MC population shifts in duodenal, ileal, and cecal sections (36.7, 55.4, and 36.2% SC, respectively). The CP-supplemented birds had higher SC between pre- and postchallenge MC in duodenal and ileal (73.3, 81.8%) than COV (66.4, 66.5%). However, COV broilers had the smallest changes in cecal MC after infection (79.5% SC). We concluded that cocci-vaccination causes small changes in intestinal MC, but challenge causes drastic shifts. The EO blend supplementation modulates MC in cocci-vaccinated broilers, avoiding drastic shifts after a mixed coccidia infection

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

    PubMed

    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.

  16. Identification and characterization of a cathepsin-L-like peptidase in Eimeria tenella.

    PubMed

    Liu, Renqiang; Ma, Xueting; Liu, Aijun; Zhang, Lei; Cai, Jianping; Wang, Ming

    2014-12-01

    Avian coccidiosis, caused by Eimeria spp., is one of the major parasitic diseases in birds. Cysteine protease is a major virulence factor in parasitic protozoa, and it may be a suitable chemotherapeutic target and vaccine candidate molecule. A 100 amino acid (aa.) partial sequence of cathepsin L, which is a cysteine protease, was reported by Katrib et al. (Ac. No. CDJ41293) (2012). A 219 aa. sequence was reported by Reid et al. (Ac. No. AFV92863) (2013). However, the open reading frame (ORF) was not reported. In this study, a full sequence of a cathepsin-L-like peptidase in Eimeria tenella (EtcatL) was obtained and its biochemical characterizations and expression profiles were analyzed across different stages of the parasite's life cycle. Results showed that the EtcatL gene encodes a protein 470 aa. in length, with 47 and 49% identity to Toxoplasma gondii and Eimeria acervulina. Considering the close phylogenetic relationship, TgcatL (PDB. ID 3F75) was selected for use as a template for homology modeling with quality factors of 90.9. Gelatin SDS-PAGE showed it to exert protease activity at ≈38 and ≈26 kDa. Further analysis showed the kinetic parameters of the recombinant peptidase to be K m  = 8.9 μM and V max = 5.7 RFU/s μM at pH 5.5 containing 10 mM dithiothreitol (DTT) in the reaction matrix, and the IC50 value of E64 was 65.32 ± 3.02 nM. The recombinant protein was active from 25 to 50 °C, with optimal activity at 42 °C. The RT-PCR and Western blot results showed it to be expressed mainly at the endogenous stages and the initial phase of the sporulation. The protective experiment showed that chickens immunized with 100 and 200 μg rEtcatL had reduction of weight loss values 48.7 and 57.9% those of infected controls, respectively. Their reduction of lesion scores (RLS) were 25.0 and 47.2% that of control chickens, and relative oocyst production (ROP) was 39.6 and 15.5% that of control chickens. These results indicate that the EtcatL can be

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

    PubMed

    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.

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

    PubMed

    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.

  19. Prevalence of Eimeria macusaniensis and Eimeria ivitaensis in South American camelids of Northwest Argentina.

    PubMed

    Cafrune, M M; Marín, R E; Rigalt, F A; Romero, S R; Aguirre, D H

    2009-06-10

    Faecal samples from mostly adult llamas (n=626), vicuñas (n=161) and guanacos (n=4) were obtained between December 2004 and July 2008 in three Provinces of Northwest Argentina in order to study the prevalence of Eimeria macusaniensis and Eimeria ivitaensis. Faeces were examined by a flotation technique using a Cl(2)Zn+ClNa solution (specific gravity=1.59). Oocysts of E. macusaniensis occurred in 88.3% of 77 llama herds and in 50.3% of 626 llamas sampled whereas oocysts of E. ivitaensis occurred in only four llamas (herd and llama prevalence of 5.2% and 0.6%, respectively). The individual prevalence of E. macusaniensis in vicuñas and guanacos were of 14.3% and 25.0%, respectively. E. ivitaensis was not detected in these latter species. The results showed a prevalence of E. macusaniensis higher than previously reported in adult domestic camelids (llamas and alpacas). In contrast, the very low prevalence of E. ivitaensis in llamas and its absence in wild camelids (vicuñas and guanacos) was remarkable. Differences between prevalence of both coccidian species are discussed.

  20. Maxima and O-C Diagrams for 489 Mira Stars

    NASA Astrophysics Data System (ADS)

    Karlsson, T.

    2013-11-01

    Maxima for 489 Mira stars have been compiled. They were computed with data from AAVSO, AFOEV, VSOLJ, and BAA-VSS and collected from published maxima. The result is presented in a mysql database and on web pages with O-C diagrams, periods and some statistical information for each star.

  1. Biochemical characterization of enoyl reductase involved in Type II fatty acid synthesis in the intestinal coccidium Eimeria tenella (Phylum Apicomplexa).

    PubMed

    Cai, Xiaomin; Lorraine Fuller, A; McDougald, Larry R; Tan, Xiangshi; Cai, Jianping; Wang, Feng; Sacchettini, James C; Zhu, Guan

    2007-07-01

    An enoyl reductase (EtENR) closely related to those of green algae and involved in Type II fatty acid synthesis was characterized and localized to the apicoplast in the coccidium Eimeria tenella. Biochemical analysis using native EtENR protein extracted from parasites confirmed its function as an enoyl reductase using NADH as a cofactor. However, the recombinant form (rEtENR) expressed in bacteria was only able to oxidize NADH, but unable to transfer the electron to enoyl-CoA, possibly due to the inappropriate folding of rEtENR expressed in bacteria. The functions of both native and recombinant EtENR could be inhibited by triclosan (IC(50)=1.45 microM), suggesting that this enzyme may be explored as a drug target against coccidiosis.

  2. High prevalence of Eimeria infection in dairy goats in Shaanxi province, northwestern China.

    PubMed

    Zhao, Guang Hui; Lei, Li-Hui; Shang, Chuan-Chuan; Gao, Man; Zhao, Yan Qing; Chen, Chao-Xi; Chen, De-Kun

    2012-06-01

    A survey of dairy goats for infection with Eimeria species of coccidia was conducted in the Shaanxi province, northwestern China between December and November 2010, including Saanen and Guanzhong breeds. A total of 584 fecal samples (250 and 334 from Saanen and Guanzhong dairy goats, respectively) in six farms were collected. Eimeria oocysts were seen in 568 (97.3%) fecal samples, with six species, namely Eimeria jolchijevi, Eimeria arloingi, Eimeria alijevi, Eimeria caprina, Eimeria hirci, and Eimeria christenseni. The most prevalent were E. arloingi in Saanen and Guanzhong dairy goats, with an overall prevalence of 83.3% and 84.4%, and the lowest prevalence were E. christenseni (26.9%) and E. hirci (20.7%) for Saanen and Guanzhong Dairy goats, respectively. Two or more Eimeria species were commonly presented in all the age groups; 80.0% and 81.4% of positive Saanen and Guanzhong dairy goats carried more than two species, and 1.6% and 6.5% of two breeds had six species. The results of the present survey suggested that Eimeria infection is wide and severe in the Saanen and Guanzhong dairy goats, which suggested that integrated strategies should be implemented to prevent and control coccidial infection in dairy goats in this province.

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

    PubMed

    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.

  4. Identification of virus-like particles in Eimeria stiedae.

    PubMed

    Revets, H; Dekegel, D; Deleersnijder, W; De Jonckheere, J; Peeters, J; Leysen, E; Hamers, R

    1989-10-01

    When nucleic acid samples purified from sporozoites of Eimeria stiedae were analyzed by agarose gel electrophoresis, an ethidium-stainable band with an apparent electrophoretic mobility of 6.5 kb was consistently observed. The band was readily degradable upon RNAse treatment, and its susceptibility towards ribonuclease A on a decreasing ionic strength was suggestive of double-stranded RNA (dsRNA). Electron microscopy revealed spherical, probably icosahedral, virus-like particles (VLP) with a diameter of 35 nm in sporozoite lysates. The VLP were purified by CsCl buoyant density gradient centrifugation. Upon extraction, these particles yielded dsRNA molecules of a uniform length of 1.63 microns. The presence of the VLP was investigated in different Eimeria strains. All E. stiedae isolates contained the RNA virus, whereas the Eimeria intestinalis and Eimeria magna isolates tested did not. RNA/RNA hybridization experiments where the E. stiedae VLP dsRNA was probed to the genomes of the dsRNA viruses of Trichomonas vaginalis and Giardia intestinalis revealed a strong relatedness of the E. stiedae virus to the G. intestinalis virus, in contrast with the T. vaginalis virus, where no homology could be detected.

  5. Molecular and morphological characterization of Eimeria in game birds.

    USDA-ARS?s Scientific Manuscript database

    Coccidiosis is generally known as a disease of economic importance in chickens. However, Eimeria species also negatively impacts the game bird industry. Here we present data generated from litter samples collected from two farms experiencing coccidiosis in chukars and pheasants. From morphological...

  6. Efficacy of diclazuril against Eimeria dispersa in turkeys.

    PubMed

    Vanparijs, O; Hermans, L; Marsboom, R

    1991-01-01

    Diclazuril, a new anticoccidial drug, was tested for its efficacy against Eimeria dispersa in turkeys. A dose-titration study indicated that diclazuril at dosages of 0.5, 1, and 2 ppm in the feed was highly effective in terms of weight gain and suppression of lesions, abnormal droppings, and oocyst shedding.

  7. Molecular characterization and phylogenetic analysis of Eimeria from turkeys and gamebirds, and the implication on the evolutionary relationships OF Eimeria FROM galliforme birds

    USDA-ARS?s Scientific Manuscript database

    In order to evaluate the evolutionary relationships of Eimeria that infect galliforme birds, the 18s rDNA gene, and a portion of the cytochrome oxidase subunit 1 (cox-1) were amplified from Eimeria that infect turkeys, chukars, and pheasants. The phylogenetic analysis of these sequences suggests th...

  8. The rhoptry proteome of Eimeria tenella sporozoites.

    PubMed

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

    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.

  9. Apicoplast genome of the coccidian Eimeria tenella.

    PubMed

    Cai, Xiaomin; Fuller, A Lorraine; McDougald, Larry R; Zhu, Guan

    2003-12-04

    Unicellular apicomplexans possess an algal-originated plastid referred to as an apicoplast. Although apicomplexan parasites are comprised of highly diverse protists, the complete apicoplast genome sequences have only been determined from the hematozoan Plasmodium falciparum and cyst-forming coccidian Toxoplasma gondii. Here, we report the third complete sequence of apicoplast genome from the intestinal coccidian Eimeria tenella that may serve as a new drug target against coccidiosis in the livestock. The AT-rich E. tenella plastid genome is a 35-kb circular element. Its gene organization resembles more closely that of T. gondii than P. falciparum. Although the E. tenella plastid genome contains an almost identical set of genes to that found in P. falciparum and T. gondii, its encoded genes share low or moderate homologies with their counterparts in the other two apicomplexans. With the addition of this coccidian plastid genome sequence, we attempted to reexamine the apicoplast genome evolution and performed phylogenetic reconstructions using maximum likelihood and Bayesian inference (BI) methods based on a concatenated dataset of plastid-encoded rpoB, rpoC1 and rpoC2 proteins. All resulting rpo protein trees placed apicoplast as a sister to Euglena within the green lineage. On the other hand, many recent studies based on the organization of plastid genes and some nuclear-encoded plastid proteins have supported a common red algal ancestry of apicomplexan and dinoflagellate plastids. If the apicoplast indeed originated from a red ancestor, the green relationship of apicomplexan genes would probably imply that the ancestral host that gave rise to the (red) apicoplast might have already contained some primary green plastid genes.

  10. Molecular characterization and analysis of a novel calcium-dependent protein kinase from Eimeria tenella.

    PubMed

    Han, H Y; Zhu, S H; Jiang, L L; Li, Y; Dong, H; Zhao, Q P; Kong, C L; Huang, B

    2013-05-01

    The calcium-dependent protein kinases (CDPKs) are unique enzymes found only in plants, green algae, ciliates and apicomplexan parasites. In this study, a novel CDPK gene of Eimeria tenella, designed EtCDPK3, was cloned using rapid amplification of cDNA ends (RACE) based on the expressed sequence tag (EST). The entire cDNA of EtCDPK3 contained 1637 nucleotides encoding 433 amino acids and the deduced EtCDPK3 protein had canonical characteristic domains identified in other CDPKs, including a well-conserved amino-terminal kinase domain and a carboxy-terminal calmodulin-like structure with 4 EF-hand motifs for calcium binding. The expression profiles of the EtCDPK3 gene in different development stages were investigated by real-time quantitative PCR. Messenger RNA levels from the EtCDPK3 gene were higher in sporozoites than in other stages (unsporulated oocysts, sporulated oocysts and merozoites). Western blot analysis showed that rabbit antiserum against recombinant EtCDPK3 could recognize a native 49 kDa protein band of parasite. Indirect immunofluorescent antibody labelling revealed dispersed localization of EtCDPK3 during the first schizogony and intense specific staining. EtCDPK3 was located at the apical end of the sporozoites after early infection of DF-1 cells and the protein was highly expressed. Inhibition of EtCDPK3 function using specific antibodies reduced the ability of E. tenella to invade host cells. These results suggested that EtCDPK3 may be involved in invasion and survival of the parasite intracellular stages of E. tenella. Because this kinase family is absent from hosts, it represents a valid target that could be exploited for chemotherapy against Eimeria spp.

  11. Protective immunity against Eimeria tenella infection in chickens following oral immunization with Bacillus subtilis expressing Eimeria tenella 3-1E protein.

    PubMed

    Lin, Zhiwei; Shi, Yanyun; Deng, Bin; Mao, Xiangfei; Yu, Dongyou; Li, Weifen

    2015-09-01

    The current experiment was conducted to construct recombinant Bacillus subtilis WB600 expressing Eimeria tenella 3-1E protein to investigate the oral immunization protective effects against E. tenella. The merozoite surface antigen 3-1E gene of E. tenella was introduced into the pBS-H1 expression vector with a novel signal peptide sequence. After the electro-transformation, the expression of objective protein in B. subtilis WB600 was detected by Western blot. The results showed that the recombinant B. subtilis strain with the ability of high-level secretion of 3-1E was constructed successfully. Seven-day-old broiler chickens were orally vaccinated with B. subtilis WB600 harboring 3-1E (B.S-pBS-H1-3-1E) or B. subtilis WB600 with empty plasmid (B.S-pBS-H1) 10 days prior to challenge with sporulated E. tenella oocysts. The results showed the recombinant B. subtilis strain with the ability of high-level secretion of 3-1E was constructed successfully. Vaccination with B.S-pBS-H1-3-1E strain significantly increased the anti-coccidial index and reduced cecal lesion scores compared with the positive control group (chickens were challenged with sporulated E. tenella oocysts without oral administration of B.S-pBS-H1-3-1E strain) and B.S-pBS-H1 group. Ceca mucosal sIgA, secretion, and IL-2, IL-12, IFN-γ, and IL-10 level after challenge were greater in the B.S-pBS-H1-3-1E group than in the positive control group. Taken together, these results indicated that B. subtilis WB600 harboring 3-1E protein induces protective immunity against E. tenella.

  12. Heat shock protein 90 genes of two species of poultry Eimeria: expression and evolutionary analysis.

    PubMed

    Miska, K B; Fetterer, R H; Min, W; Lillehoj, H S

    2005-04-01

    Heat shock protein 90 (Hsp90) is 1 of the most abundant and evolutionarily conserved proteins. In most species, Hsp90 is essential for proper cell function. In this study, we present the molecular analysis of Hsp90 from Eimeria species, the causative agents of avian coccidiosis. The full-length Eimeria acervulina Hsp90 complementary DNA was isolated from intestinal intraepithelial lymphocytes of Eimeria-infected chickens. From evolutionary analysis and sequence identity, it is likely that Eimeria Hsp90 sequences described thus far encode the cytosolic versions of the protein. Although at the nucleotide and amino acid levels Eimeria tenella and E. acervulina Hsp90 are highly similar, their expression profiles differ considerably. Although E. tenella transcripts were detected in all developmental stages tested, E. acervulina transcripts were not found in oocysts undergoing sporulation or in fully sporulated oocysts, suggesting that messenger RNA expression may be regulated quite differently between Eimeria species.

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

    PubMed

    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 < 0.05) and comparable to that of the lasalocid-treated birds in all 3 trials. Mean BW of chicks in all treatment groups decreased uniformly as the litter aged and moisture content increased. The mean intestinal coliform population from YCR-treated chicks was lower (P < 0.05) than those of the control and lasalocid populations. The coliform count was consistently lower than that in chicks on a bacitracin-supplemented diet. Coliform counts from the control and lasalocid-treated birds did not differ. The 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.

  14. The effect of infection with mixed Eimeria species on hematology and immune responses following Newcastle disease and infectious bursal disease booster vaccination in broilers.

    PubMed

    Akhtar, Masood; Awais, Mian Muhammad; Anwar, Muhammad Irfan; Ehtisham-ul-Haque, Syed; Nasir, Amar; Saleemi, Muhammad Kashif; Ashraf, Kamran

    2015-03-01

    Coccidiosis is an important parasitic disease of chickens, causing high mortality and morbidity. This morbidity is believed to be correlated with altered population dynamics of blood cells and immunocompromisation. This study investigated the effects of mixed Eimeria species (viz., tenella, maxima, acervulina and necatrix) infection on hematology and immune responses following Newcastle disease (ND) and infectious bursal disease (IBD) booster vaccination in broilers. One-day-old broiler chicks (Hubbard; n = 200) were divided into two equal groups A and B. On day 16, group A was infected orally with Eimeria species (7 × 10(4) sporulated oocysts), whereas group B served as control. Both groups were analyzed for hematological parameters on post-infection days 6-8. Sera from both groups were analyzed for antibody titers against ND and IBD vaccines. On day 8 post-infection, lymphoid organs were also examined. Significantly lower (P < 0.05) levels of plasma proteins, globular volume, hemoglobin concentration, packed cell volume, total erythrocytes, mean corpuscular volume and mean corpuscular hemoglobin concentration were found in infected chickens compared with non-infected control chickens. In addition, the infected group exhibited significantly increased (P < 0.05) numbers of different leukocytes. Infected chickens also showed significantly lower antibody titers against ND and IBD with decreased relative organ weights of all lymphoid organs except spleen. Mixed species of Eimeria adversely affected the hematology and immune efficiency of broilers. Thus, inexpensive immune potentiators and hemotonics along with appropriate anti-coccidial medications are suggested to avoid the complications and subsequent economic losses.

  15. Subchronic toxicity study in mice fed Spirulina maxima.

    PubMed

    Salazar, M; Martínez, E; Madrigal, E; Ruiz, L E; Chamorro, G A

    1998-10-01

    The purpose of this study was to evaluate the toxicity of Spirulina maxima, a blue-green alga used as food supplement and food coloring, after 13 weeks of treatment. Groups of ten mice of each sex were given S. maxima in the diet at concentrations of 0 (control), 10, 20 or 30% (w/w) for 13 weeks. The alga ingestion had no effect on behavior, food and water intake, growth or survival. Terminal values in hematology and clinical chemistry did not reveal differences between treated and control groups. However, male and female mice showed significant changes in serum cholesterol levels at 20 and 30% algal concentrations, but a toxic effect of S. maxima was excluded. Post-mortem examination revealed no differences in gross or microscopic findings. Our results show that S. maxima up to high feeding levels did not produce adverse effects in mice after subchronic treatment.

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

    PubMed

    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.

  17. Molecular detection of field isolates of Turkey Eimeria by polymerase chain reaction amplification of the cytochrome c oxidase I gene.

    PubMed

    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.

  18. Analysis of chicken cytokine and chemokine gene expression following Eimeria acervulina and Eimeria tenella infections.

    PubMed

    Hong, Yeong Ho; Lillehoj, Hyun S; Lee, Sung Hyen; Dalloul, Rami A; Lillehoj, Erik P

    2006-12-15

    The expression levels of mRNA encoding a panel of 28 chicken cytokines and chemokines were quantified in intestinal lymphocytes following Eimeria acervulina and Eimeria tenella primary and secondary infections. Compared with uninfected controls, transcripts of the pro-inflammatory cytokines IFN-alpha, IL-1beta, IL-6, and IL-17 were increased up to 2020-fold following primary infection. By contrast, following secondary infection by either microorganism, pro-inflammatory mRNAs levels were relatively unchanged (< or = 20-fold). Transcripts encoding the Th1 and Th1 regulatory cytokines IFN-gamma, IL-2, IL-10, IL-12, IL-15, IL-16, and IL-18 were uniformly increased 14-2471-fold after E. acervulina primary infection, but either unchanged (IL-15, IL-16, IL-18), increased (IFN-gamma, IL-10, IL-12), or decreased (IL-2) following E. tenella primary infection. Following secondary infections, Th1 cytokine mRNA levels were relatively unchanged, with the exception of IL-12 which was increased 1.5 x 10(5)-fold after E. acervulina and decreased 5.1 x 10(4)-fold after E. tenella infection. Transcripts for the Th2 or Th2 regulatory cytokines IL-3 and GM-CSF were increased up to 327-fold following primary or secondary infection with both parasites, while IL-4 and IL-13 mRNAs were decreased 25- to 2 x 10(5)-fold after primary or secondary infection. The dynamics of chicken chemokine expression revealed modest changes (<100-fold) following primary or secondary infection except for lymphotactin. When lymphocyte subpopulations were similarly analyzed, IFN-gamma, IL-2, IL-3, IL-15, and MIF were most highly increased in TCR2(+) cells following E. acervulina infection, while TCR1(+) cells only expressed high levels of IL-16 following E. tenella infection. In contrast, CD4(+) cells only expressed highest levels of IL-10 after E. acervulina infection, whereas these cells produced abundant transcripts for IFN-gamma, IL-3, IL-15, and MIF after E. tenella infection. We conclude that coccidiosis

  19. Securing poultry production from the ever-present Eimeria challenge.

    PubMed

    Blake, Damer P; Tomley, Fiona M

    2014-01-01

    The intestinal disease coccidiosis, caused by protozoan parasites of the genus Eimeria, is one of the most important livestock diseases in the world. It has a high impact in the poultry industry where parasite transmission is favoured by high-density housing of large numbers of susceptible birds. Coccidiosis control in poultry is achieved by careful husbandry combined with in-feed anticoccidial drugs or vaccination with live parasites. However, outbreaks of coccidiosis still occur and subclinical infections, which significantly impact on productivity and food security, are common due to widespread drug resistance, high parasite prevalence, and environmental persistence. Herein, we review some recent approaches for the production of cheaper third generation vaccines, based on robust methods for identification of immunoprotective antigens and the use of transgenic Eimeria. Copyright © 2013 Elsevier Ltd. All rights reserved.

  20. Enzymes as feed additive to aid in responses against Eimeria species in coccidia-vaccinated broilers fed corn-soybean meal diets with different protein levels.

    PubMed

    Parker, J; Oviedo-Rondón, E O; Clack, B A; Clemente-Hernández, S; Osborne, J; Remus, J C; Kettunen, H; Mäkivuokko, H; Pierson, E M

    2007-04-01

    This research aimed to evaluate the effects of adding a combination of exogenous enzymes to starter diets varying in protein content and fed to broilers vaccinated at day of hatch with live oocysts and then challenged with mixed Eimeria spp. Five hundred four 1-d-old male Cobb-500 chickens were distributed in 72 cages. The design consisted of 12 treatments. Three anticoccidial control programs [ionophore (IO), coccidian vaccine (COV), and coccidia-vaccine + enzymes (COV + EC)] were evaluated under 3 CP levels (19, 21, and 23%), and 3 unmedicated-uninfected (UU) negative controls were included for each one of the protein levels. All chickens except those in unmedicated-uninfected negative controls were infected at 17 d of age with a mixed oral inoculum of Eimeria acervulina, Eimeria maxima, and Eimeria tenella. Live performance, lesion scores, oocyst counts, and samples for gut microflora profiles were evaluated 7 d postinfection. Ileal digestibility of amino acids (IDAA) was determined 8 d postinfection. Microbial communities (MC) were analyzed by G + C%, microbial numbers were counted by flow cytometry, and IgA concentrations were measured by ELISA. The lowest CP diets had poorer (P < or = 0.001) BW gain and feed conversion ratio in the preinfection period. Coccidia-vaccinated broilers had lower performance than the ones fed ionophore diets during pre- and postchallenge periods. Intestinal lesion scores were affected (P < or = 0.05) by anticoccidial control programs, but responses changed according to gut section. Feed additives or vaccination had no effect (P > or = 0.05) on IDAA, and diets with 23% CP had the lowest (P < or = 0.001) IDAA. Coccidial infection had no effect on MC numbers in the ileum but reduced MC numbers in ceca and suppressed ileal IgA production. The COV + EC treatment modulated MC during mixed coccidiosis infection but did not significantly improve chicken performance. Results indicated that feed enzymes may be used to modulate the gut

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

    PubMed

    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.

  2. A toolbox facilitating stable transfection of Eimeria species.

    PubMed

    Clark, Julie D; Billington, Karen; Bumstead, Janene M; Oakes, Richard D; Soon, Puay Eng; Sopp, Paul; Tomley, Fiona M; Blake, Damer P

    2008-11-01

    Stable transfection of Eimeria species has been difficult to achieve because of the obligate requirement for in vivo amplification and selection of the parasites. Strategies to generate and stabilise populations of transfected Eimeria tenella are described here, together with the identification of optimal parameters for the transfection process. A series of plasmids expressing selectable markers, including a panel of fluorescent reporter genes and a mutant Toxoplasma gondii dihydrofolate reductase-thymidylate synthase (DHFR-TSm2m3) gene that confers resistance to pyrimethamine, were electroporated into sporozoites of the E. tenella Wisconsin strain and stabilised by selective passage through chickens. Very high transfection efficiencies of up to 25% sporozoites in transient transfection and up to 9% oocysts following a single round of in vivo selection were achieved. Crucial factors include the use of very freshly harvested parasites with the AMAXA nucleofection system (program U33 in a cytomix-buffered reaction) and linearised plasmid DNA. The use of a restriction enzyme mediated integration (REMI) protocol boosted overall efficiency and elevated insertion rate per genome. Successful development of methods to generate and isolate stable populations of transfected Eimeria parasites will now stimulate rapid expansion of reverse genetic studies in this important coccidian.

  3. Prevalence and cross-immunity of Eimeria species on Korean chicken farms

    USDA-ARS?s Scientific Manuscript database

    The epidemiology of Eimeria species in poultry flocks is important to increase the effectiveness of vaccinations and prophylactic strategies on chicken farms. In this study, fecal samples from 356 chicken farms were collected randomly and examined for the prevalence of Eimeria species. Through micro...

  4. Eimeria species in wild rabbits (Oryctolagus cuniculus) in Fars province, Iran.

    PubMed

    Razavi, S M; Oryan, A; Rakhshandehroo, E; Moshiri, A; Mootabi Alavi, A

    2010-12-01

    This investigation was accomplished during February to November 2008. A total of 71 wild rabbits (Oryctolagus cuniculus) of about 5 to 12 months age were collected alive from different parts of Fars province, south of Iran. Faecal sampling was carried out directly from recti and the oocysts were isolated using sedimentation and floatation techniques and the sporulated oocyst were identified based on morphological and biological characteristics. All the rabbits were apparently healthy and showed no clinical symptoms. Twenty two rabbits (31.0%) were positive for infection with Eimeria and six species including Eimeria perforans (18.3%), Eimeria magna (16.9%), Eimeria media (14.1), Eimeria irresidua (11.2%), Eimeria flavescens (4.2%), and Eimeria coecicola (2.8%) were identified. Eighty six percent of the infected rabbits showed mixed infections with two or three Eimeria species. Lack of clinical signs could be due to the agro ecological and environmental conditions of rabbit habitats specifically dry and hot climatic features in recent years. In addition, immunity induced by long term exposure to low doses of oocysts shedded by the carrier animals probably have pivotal role in impairing parasitic developmental cycles and preventing acute coccidiosis.

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

    PubMed

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

    2008-02-02

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

  6. A rapid method for determining salinomycin and monensin sensitivity in Eimeria tenella

    USDA-ARS?s Scientific Manuscript database

    Standard methods of determining the ionophore sensitivity of Eimeria rely on infecting chickens with an isolate or a mixture of Eimeria spp. oocysts in the presence of different anti-coccidial drugs. The purpose of this study was to develop a rapid in vitro method for assessing salinomycin and mone...

  7. Pyrosequencing analysis of Salinomycin and Eimeria spp. challenge-induced changes in broiler cecal microbial communities

    USDA-ARS?s Scientific Manuscript database

    Eimeria spp. invade and damage the intestinal cell lining of broilers, resulting in cell necrosis and secondary bacterial infections. The current work investigates the effect of anticoccidial agents, salinomycin in combination with roxarsone, and Eimeria challenge on the composition of broiler ceca...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    PubMed

    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.

  10. Control of clinical coccidiosis of calves due to Eimeria bovis and Eimeria zuernii with toltrazuril under field conditions.

    PubMed

    Mundt, H-C; Bangoura, B; Mengel, H; Keidel, J; Daugschies, A

    2005-10-01

    The efficacy of metaphylactic treatment with toltrazuril (Baycox 5% suspension) against natural infections with Eimeria bovis and/or Eimeria zuernii in calves was investigated. The study was conducted with 208 calves on five calf-rearing farms in Germany and the Czech Republic. All participating farms had a notable incidence of coccidiosis. Animals were treated 14 days after stabling in the respective facility. One group was treated with 15 mg toltrazuril/kg body weight, and a second group served as the sham-treated control. Assessment of efficacy was based on faecal consistency and oocyst excretion of E. bovis and E. zuernii, both investigated throughout the study. Duration and rate of oocyst excretion as well as number of scour days with E. bovis or E. zuernii oocyst shedding and the severity of diarrhoea were significantly lower in the toltrazuril-treated groups. It is concluded that a single metaphylactic treatment with toltrazuril controls coccidiosis of housed calves under various field conditions.

  11. The Molecular Characterization and Immunity Identification of Microneme 3 of Eimeria acervulina.

    PubMed

    Zhang, ZhenChao; Liu, XinChao; Yang, XinChao; Liu, LianRui; Wang, Shuai; Lu, MingMin; Ehsan, Mohammad; Gadahi, Javaid A; Song, XiaoKai; Xu, LiXin; Yan, RuoFeng; Li, XiangRui

    2016-11-01

    The gene of Eimeria acervulina microneme protein 3 (EaMIC3) was cloned and characterized. According to the conserved sequence, the 3'- and 5'-ends of EaMIC3 were amplified by the rapid amplification of cDNA ends (RACE). The full length cDNA of this gene was obtained by overlapping the sequences of 3'- and 5'-extremities and amplification by reverse transcription PCR. The sequence analysis revealed that the opening reading frame (ORF) of EaMIC3 was 2,607 bp and encoded a protein of 868 amino acids with 93.04 kDa. Western blotting assay showed that the recombinant protein was successfully recognized by the sera of chickens experimentally infected with E. acervulina, whereas the native protein in the somatic extract of sporozoites was as well detected by sera from rats immunized with the recombinant protein of EaMIC3. Immunofluorescence analysis indicated that EaMIC3 was expressed in the sporozoites and merozoites stages of E. acervulina. Animal challenge experiments demonstrated that the recombinant protein of EaMIC3 could significantly increase the average body weight gains, decrease the mean lesion scores and the oocyst outputs of the immunized chickens and presented anticoccidial index (ACI) more than 165. Moreover, EaMIC3 protein produced significantly high level of IgG antibody, IFN-γ, IL-10, IL-17 TGF-β, CD4(+) , and CD8(+) . © 2016 The Author(s) Journal of Eukaryotic Microbiology © 2016 International Society of Protistologists.

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

    PubMed Central

    Zhai, Qi; Huang, Bing; Dong, Hui; Zhao, Qiping; Zhu, Shunhai; Liang, Siting; Li, Sha; Yang, Sihan; Han, Hongyu

    2016-01-01

    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. PMID:27309852

  13. Giant planet formation at the pressure maxima of protoplanetary disks

    NASA Astrophysics Data System (ADS)

    Guilera, O. M.; Sándor, Zs.

    2017-07-01

    Context. In the classical core-accretion planet-formation scenario, rapid inward migration and accretion timescales of kilometer size planetesimals may not favor the formation of massive cores of giant planets before the dissipation of protoplanetary disks. On the other hand, the existence of pressure maxima in the disk could act as migration traps and locations for solid material accumulation, favoring the formation of massive cores. Aims: We aim to study the radial drift of pebbles and planetesimals and planet migration at pressure maxima in a protoplanetary disk and their implications for the formation of massive cores as triggering a gaseous runaway accretion phase. Methods: The time evolution of a viscosity driven accretion disk is solved numerically introducing a a dead zone as a low-viscosity region in the protoplanetary disk. A population of pebbles and planetesimals evolving by radial drift and accretion by the planets is also considered. Finally, the embryos embedded in the disk grow by the simultaneous accretion of pebbles, planetesimals, and the surrounding gas. Results: Our simulations show that the pressure maxima generated at the edges of the low-viscosity region of the disk act as planet migration traps, and that the pebble and planetesimal surface densities are significantly increased due to the radial drift towards pressure maxima locations. However, our simulations also show that migration-trap locations and solid-material-accumulation locations are not exactly at the same positions. Thus, a planet's semi-major axis oscillations around zero torque locations predicted by MHD and HD simulations are needed for the planet to accrete all the available material accumulated at the pressure maxima. Conclusions: Pressure maxima generated at the edges of a low-viscosity region of a protoplanetary disk seem to be preferential locations for the formation and trap of massive cores.

  14. Novel Eimeria sp. isolated from a King's skink (Egernia kingii) in Western Australia.

    PubMed

    Yang, Rongchang; Brice, Belinda; Bennett, Mark D; Eliott, Aileen; Ryan, Una

    2013-02-01

    A novel Eimeria sp. was identified in faeces collected from a King's skink (Egernia kingii) housed at the Kanyana Wildlife Rehabilitation Centre in Western Australia. Oocysts measure 17.0×15.0 μm with a length/width ratio (L/W) of 1.13. Phylogenetic analysis of 18S rRNA sequences indicated that the novel Eimeria sp. shared the highest genetic similarity to Eimeria antrozoi and Eimeria rioarribaensis from vespertilionid bats from North America (≥98.9%). At the COI locus, bat-derived sequences were not available and phylogenetic analysis placed the novel Eimeria sp. in a clade by itself and shared 98.8% similarity with the rodent-derived species E. falciformis and E. vermiformis. This suggests that the isolate from the King's skink's faeces was probably derived from a mammal, possibly a rodent or a bat.

  15. The cosmic microwave background after MAXIMA and BOOMERANG

    SciTech Connect

    Richards, Paul L.

    2000-12-01

    The first release of data from the MAXIMA and BOOMERANG experiments has introduced a new era of precision cosmology. The two data sets are essentially independent, consistent and complementary. In a joint effort by the two teams, the two data sets were combined and then used to test cosmological models and determine values of cosmological constants. These results are available because of the success of bolometric detection techniques. The experimental approach is described with references to the MAXIMA-1 experiment. Important new cosmological experiments at far infrared and millimeter wavelengths require major improvements in bolometric techniques. A new technology, the voltage-biased superconducting bolometer, promises to provide the required experimental power.

  16. Eimeria that infect fish are diverse and are related to, but distinct from, those that infect terrestrial vertebrates.

    PubMed

    Molnár, Kálmán; Ostoros, Györgyi; Dunams-Morel, Detiger; Rosenthal, Benjamin M

    2012-12-01

    The Eimeria are ubiquitous parasites (Phylum: Apicomplexa; family: Coccidia) of the gut epithelium of vertebrates which complete their development in a single host species and whose sporocysts may be recognized by the presence of a Stieda body through which their sporozoites excyst. Their diversity and relationship to other kinds of coccidia have been successfully explored by molecular systematic studies based on the sequencing the 18S ribosomal DNA. To date, most attention has been paid to the diversity and evolutionary relationships of Eimeria spp. parasitizing terrestrial vertebrates, most especially those species infecting domesticated birds and mammals. Regrettably, no Eimeria have yet been considered from the Earth's first vertebrates: the fish. If Eimeria first evolved in fish, then extant piscine parasites should comprise a deeply branching assemblage at the base of well-constructed phylogenetic trees. Here, we sequenced portions of ribosomal DNA from several such isolates (from Eimeria anguillae, Eimeria daviesae, Eimeria percae, Eimeria variabilis, Eimeria rutili and Eimeria nemethi) and compared them to one another as well as to other available sequences from the parasites of fish and terrestrial vertebrates, in order to better understand their diversity and origins. By establishing that such piscine parasites comprise a deeply branching clade at the base of the Eimeriidae, these data substantiate the hypothesis that Eimeria may have originated in fish. Plainly, a great deal of coccidian diversity awaits future discovery and description.

  17. 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.

    PubMed

    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.

  18. Transgenic Eimeria tenella as a vaccine vehicle: expressing TgSAG1 elicits protective immunity against Toxoplasma gondii infections in chickens and mice

    PubMed Central

    Tang, Xinming; Yin, Guangwen; Qin, Mei; Tao, Geru; Suo, Jingxia; Liu, Xianyong; Suo, Xun

    2016-01-01

    The surface antigen 1 of Toxoplasma gondii (TgSAG1) is a major immunodominant antigen and is widely considered an ideal candidate for the development of an effective recombinant vaccine against toxoplasmosis. Eimeria tenella, an affinis apicomplexan parasite with T. gondii, is a potential vaccine vector carrying exogenous antigens that stimulates specific immune responses. Here, we engineered TgSAG1 into E. tenella and obtained a stably transfected E. tenella line (Et-TgSAG1). We found TgSAG1 localized on the cell surface of Et-TgSAG1, which is similar to its native distribution in T. gondii tachyzoites. We immunized the chickens with Et-TgSAG1 orally and detected TgSAG1-specific immune responses, which partly reduced T. gondii infection. In the mouse model, we immunized the mice with Et-TgSAG1 sporozoites intraperitoneally and challenged them with T. gondii tachyzoites RH strain. We found that the mice immunized with Et-TgSAG1 showed a TgSAG1 specific Th 1-dominant immune response and a prolonged survival time compared with wild-type E. tenella and non-immunized mice. Collectively, our results demonstrated that Et-TgSAG1, utilized as a recombinant vaccine against toxoplasmosis, could be applied in both chickens and mice. Our findings also provide a promising persuasion for the development of transgenic Eimeria as vaccine vectors for use in birds and mammals. PMID:27387302

  19. Temperature maxima in stable two-dimensional shock waves

    NASA Astrophysics Data System (ADS)

    Kum, Oyeon; Hoover, Wm. G.; Hoover, C. G.

    1997-07-01

    We use molecular dynamics to study the structure of moderately strong shock waves in dense two-dimensional fluids, using Lucy's pair potential. The stationary profiles show relatively broad temperature maxima, for both the longitudinal and the average kinetic temperatures, just as does Mott-Smith's model for strong shock waves in dilute three-dimensional gases.

  20. Scilab and Maxima Environment: Towards Free Software in Numerical Analysis

    ERIC Educational Resources Information Center

    Mora, Angel; Galan, Jose Luis; Aguilera, Gabriel; Fernandez, Alvaro; Merida, Enrique; Rodriguez, Pedro

    2010-01-01

    In this work we will present the ScilabUMA environment we have developed as an alternative to Matlab. This environment connects Scilab (for numerical analysis) and Maxima (for symbolic computations). Furthermore, the developed interface is, in our opinion at least, as powerful as the interface of Matlab. (Contains 3 figures.)

  1. Scilab and Maxima Environment: Towards Free Software in Numerical Analysis

    ERIC Educational Resources Information Center

    Mora, Angel; Galan, Jose Luis; Aguilera, Gabriel; Fernandez, Alvaro; Merida, Enrique; Rodriguez, Pedro

    2010-01-01

    In this work we will present the ScilabUMA environment we have developed as an alternative to Matlab. This environment connects Scilab (for numerical analysis) and Maxima (for symbolic computations). Furthermore, the developed interface is, in our opinion at least, as powerful as the interface of Matlab. (Contains 3 figures.)

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

    PubMed

    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 < 0.001). 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

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

    PubMed Central

    2013-01-01

    Background 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. Results 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 < 0.001). 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

  4. Molecular cloning and characterization of lactate dehydrogenase gene from Eimeria tenella.

    PubMed

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

    2014-08-01

    Lactate dehydrogenase (LDH) is a key enzyme in the glycolytic pathway and is crucial for parasite survival. In this study, we cloned and expressed the LDH of Eimeria tenella (EtLDH). Real-time polymerase chain reaction and Western blot analysis revealed that the expression of EtLDH was developmentally regulated at the messenger RNA (mRNA) and protein levels. EtLDH mRNA levels were higher in second-generation merozoites than in other developmental stages (unsporulated oocysts, sporulated oocysts, and sporozoites). EtLDH protein expression levels were most prominent in second-generation merozoites, moderately expressed in unsporulated oocysts and sporulated oocysts, and weakly detected in sporozoites. Immunostaining with anti-recombinant EtLDH (rEtLDH) antibody indicated that EtLDH was mainly located in the anterior region in free sporozoites and became concentrated in the anterior region of intracellular sporozoites except for the apex after invasion into DF-1 cells. Specific staining of EtLDH protein was more intense in trophozoites and immature first-generation schizonts, but decreased in mature first-generation schizonts. Inhibition of EtLDH function using specific antibodies cannot efficiently reduce the ability of E. tenella sporozoites to invade host cells. These results suggest that EtLDH may be involved in glycolysis during the first-generation merogony stage in E. tenella and has little role in host invasion.

  5. Functional characterizations of malonyl-CoA:acyl carrier protein transacylase (MCAT) in Eimeria tenella.

    PubMed

    Sun, Mingfei; Zhu, Guan; Qin, Zonghua; Wu, Caiyan; Lv, Minna; Liao, Shenquan; Qi, Nanshan; Xie, Mingquan; Cai, Jianping

    2012-07-01

    Eimeria tenella, an apicomplexan parasite in chickens, possesses an apicoplast and its associated metabolic pathways including the Type II fatty acid synthesis (FAS II). Malonyl-CoA:acyl-carry protein transacylase (MCAT) encoded by the fabD gene is one of the essential enzymes in the FAS II system. In the present study, the entire E. tenella MCAT gene (EtfabD) was cloned and sequenced. Immunolabeling located this protein in the apicoplast organelle in coccidial sporozoites. Functional replacement of the fabD gene with amber mutation of E. coli temperature-sensitive LA2-89 strain by E. tenella EtMCAT demonstrated that EcFabD and EtMCAT perform the same biochemical function. The recombinant EtMCAT protein was expressed and its general biochemical features were also determined. An alkaloid natural product corytuberine (CAS: 517-56-6) could specifically inhibit the EtMCAT activity (IC(50)=16.47μM), but the inhibition of parasite growth in vitro by corytuberine was very weak (the predicted MIC(50)=0.65mM).

  6. Identification of a telomeric DNA-binding protein in Eimeria tenella.

    PubMed

    Zhao, Na; Gong, Pengtao; Li, Zhipeng; Cheng, Baiqi; Li, Jianhua; Yang, Zhengtao; Li, He; Yang, Ju; Zhang, Guocai; Zhang, Xichen

    2014-09-05

    Coccidiosis is considered to be a major problem for the poultry industry, and coccidiosis control is yet urgent. Due to the roles in telomere length regulation and end protection, telomere-binding proteins have been considered as a good target for drug design. In this work, a putative Gbp1p that is similar to telomeric DNA-binding protein Gbp (G-strand binding protein) of Cryptosporidium parvum, was searched in the database of Eimeria tenella. Sequence analysis indicated E.tenella Gbp1p (EtGbp1p) has significant sequence similarity to other eukaryotic Gbps in their RNA recognition motif (RRM) domains. Electrophoretic mobility shift assays (EMSAs) demonstrated recombinant EtGbp1p bound G-rich telomeric DNA, but not C-rich or double-stranded telomeric DNA sequences. Competition and antibody supershift assays confirmed the interaction of DNA-protein complex. Chromatin immunoprecipitation assays confirmed that EtGbp1p interacted with telomeric DNA in vivo. Collectively, these evidences suggest that EtGbp1p represents a G-rich single-stranded telomeric DNA-binding protein in E.tenella.

  7. Protective immunity induced by a DNA vaccine encoding Eimeria tenella rhomboid against homologous challenge.

    PubMed

    Liu, Yingli; Zheng, Jun; Li, Jianhua; Gong, Pengtao; Zhang, Xichen

    2013-01-01

    Rhomboid protein in Apicomplexa was associated with the process of host cell invasion. To evaluate the potential of the protein in eliciting protective immunity against challenge, a DNA vaccine pVAX1-Rho encoding Eimeria tenella rhomboid was constructed. Recombinant protein was expressed in Hela cells and verified by indirect immunofluorescence and western blotting analysis. In vivo experiments, 1-week-old chickens were randomly divided into three groups. Experimental group of chickens were immunized with DNA vaccines while control group of chickens were injected with pVAX1 plasmid alone or sterile water. Two weeks following the booster dose, all chickens were inoculated orally with 5 × 10(4) sporulated oocysts of E. tenella. The host immunity and protective efficacy of this vaccine against E. tenella challenge in broilers were evaluated. Results showed that specific antibody, the levels of interleukin-2 (IL-2), interferon-γ (IFN-γ), and the percentages of CD4(+) and CD8(+) T lymphocyte cells were significantly increased in the pVAX1-Rho group. Challenge experiments demonstrated that pVAX1-Rho vaccination could reduce oocyst excretion, decrease cecal lesion, increase bodyweight gains and provide chickens with oocysts decrease ratio around 75.8 %. These results suggest that the pVAX1-Rho was able to induce humoral and cellular responses and generate protective immunity against E. tenella infection.

  8. In vivo role of tumor necrosis-like factor in Eimeria tenella infection.

    PubMed

    Zhang, S; Lillehoj, H S; Ruff, M D

    1995-01-01

    The effect of tumor necrosis-like factor (TNLF) on the pathogenesis of coccidiosis was investigated. Injection of crude chicken TNLF enhanced the weight loss caused by Eimeria tenella infection. Rabbit polyclonal antibody against recombinant human tumor necrosis factor-alpha (rhTNF) partially restored E. tenella-induced weight loss in SC chickens, but not in TK chickens. However, injection of chickens with chicken TNLF, rhTNF, and rabbit serum against rhTNF had no significant effect on cecal lesions. Both SC and TK chickens produced circulating TNLF following primary, but not secondary infection, and SC chickens showed higher level of TNLF production than TK chickens. Peripheral blood leukocyte-derived macrophages from SC and TK chickens produced a significant amount of TNLF compared to the preinfection condition when cocultured with sporozoites. In general, macrophages from SC chickens produced higher levels of TNLF than those from TK chickens. No significant difference was observed between primary and secondary infection. These results suggest that the excessive TNF production may be involved in weight loss caused by E. tenella infection in SC chickens.

  9. An optimised protocol for molecular identification of Eimeria from chickens.

    PubMed

    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

    2014-01-17

    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.

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

    PubMed Central

    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

  11. Eimeria macusaniensis associated lesions in neonate alpacas dying from enterotoxemia.

    PubMed

    Rosadio, R; Londoñe, P; Pérez, D; Castillo, H; Véliz, A; Llanco, L; Yaya, K; Maturrano, L

    2010-02-26

    Histopathological analysis of 108 intestine samples (103 grossly affected ileum and 5 jejunum) taken from Clostridium-induced neonatal alpaca (Vicugna pacos) enterotoxemia mortalities collected in the Departments of Arequipa, Puno and Cusco of southern Peru during the 2005-2008 birth seasons (January-March), revealed the presence of large numbers of both asexual and sexual stages of Eimeria macusaniensis in 33/108 (30.55%) of the samples with moderate to severe necrotized and/or hemorrhagic enteritis. It is proposed that damage to the mucosa produced by coccidial infections may facilitate overgrowth of Clostridium perfringens with toxin production leading to fatal enterotoxemia.

  12. Sequence-based genotyping clarifies conflicting historical morphometric and biological data for 5 Eimeria species infecting turkeys.

    PubMed

    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

  13. Critical heat flux maxima during boiling crisis on textured surfaces

    NASA Astrophysics Data System (ADS)

    Dhillon, Navdeep Singh; Buongiorno, Jacopo; Varanasi, Kripa K.

    2015-09-01

    Enhancing the critical heat flux (CHF) of industrial boilers by surface texturing can lead to substantial energy savings and global reduction in greenhouse gas emissions, but fundamentally this phenomenon is not well understood. Prior studies on boiling crisis indicate that CHF monotonically increases with increasing texture density. Here we report on the existence of maxima in CHF enhancement at intermediate texture density using measurements on parametrically designed plain and nano-textured micropillar surfaces. Using high-speed optical and infrared imaging, we study the dynamics of dry spot heating and rewetting phenomena and reveal that the dry spot heating timescale is of the same order as that of the gravity and liquid imbibition-induced dry spot rewetting timescale. Based on these insights, we develop a coupled thermal-hydraulic model that relates CHF enhancement to rewetting of a hot dry spot on the boiling surface, thereby revealing the mechanism governing the hitherto unknown CHF enhancement maxima.

  14. Biogas production from anaerobic digestion of Spirulina maxima algal biomass

    SciTech Connect

    Samson, R.; LeDuy, A.

    1982-08-01

    The semimicroscopic blue-green alga Spirulina maxima makes an ideal substrate for anaerobic digestion because it is easy to harvest, it can use carbon dioxide from the atmosphere as its carbon source, and its fermentability is higher than that of other small algae. Digestion experiments demonstrated that S. maxima can serve as the sole nutrient for biogas production and that municipal sewage sludge, when adapted to this new substrate, is very stable. During semicontinuous daily-fed trials under non-optimal conditions at an 0.06 lb volatile solids (VS)/ft/sup 3/ (0.97 kg VS/m/sup 3/) loading rate, 33-day retention time, and 86/sup 0/F (30/sup 0/C) digestion temperature, the daily methane yield was 4.2 CF/lb (0.26 m/sup 3//kg) VS added, which represents 47% of the maximum theoretical yield. Studies on optimizing the process are underway.

  15. Critical heat flux maxima during boiling crisis on textured surfaces.

    PubMed

    Dhillon, Navdeep Singh; Buongiorno, Jacopo; Varanasi, Kripa K

    2015-09-08

    Enhancing the critical heat flux (CHF) of industrial boilers by surface texturing can lead to substantial energy savings and global reduction in greenhouse gas emissions, but fundamentally this phenomenon is not well understood. Prior studies on boiling crisis indicate that CHF monotonically increases with increasing texture density. Here we report on the existence of maxima in CHF enhancement at intermediate texture density using measurements on parametrically designed plain and nano-textured micropillar surfaces. Using high-speed optical and infrared imaging, we study the dynamics of dry spot heating and rewetting phenomena and reveal that the dry spot heating timescale is of the same order as that of the gravity and liquid imbibition-induced dry spot rewetting timescale. Based on these insights, we develop a coupled thermal-hydraulic model that relates CHF enhancement to rewetting of a hot dry spot on the boiling surface, thereby revealing the mechanism governing the hitherto unknown CHF enhancement maxima.

  16. Critical heat flux maxima during boiling crisis on textured surfaces

    PubMed Central

    Dhillon, Navdeep Singh; Buongiorno, Jacopo; Varanasi, Kripa K.

    2015-01-01

    Enhancing the critical heat flux (CHF) of industrial boilers by surface texturing can lead to substantial energy savings and global reduction in greenhouse gas emissions, but fundamentally this phenomenon is not well understood. Prior studies on boiling crisis indicate that CHF monotonically increases with increasing texture density. Here we report on the existence of maxima in CHF enhancement at intermediate texture density using measurements on parametrically designed plain and nano-textured micropillar surfaces. Using high-speed optical and infrared imaging, we study the dynamics of dry spot heating and rewetting phenomena and reveal that the dry spot heating timescale is of the same order as that of the gravity and liquid imbibition-induced dry spot rewetting timescale. Based on these insights, we develop a coupled thermal-hydraulic model that relates CHF enhancement to rewetting of a hot dry spot on the boiling surface, thereby revealing the mechanism governing the hitherto unknown CHF enhancement maxima. PMID:26346098

  17. Histopathology of oedema in pearl oysters Pinctada maxima.

    PubMed

    Jones, J B; Crockford, M; Creeper, J; Stephens, F

    2010-07-26

    In October 2006, severe mortalities (80 to 100%) were reported in pearl oyster Pinctada maxima production farms from Exmouth Gulf, Western Australia. Only P. maxima were affected; other bivalves including black pearl oysters P. margaratifera remained healthy. Initial investigations indicated that the mortality was due to an infectious process, although no disease agent has yet been identified. Gross appearance of affected oysters showed mild oedema, retraction of the mantle, weakness and death. Histology revealed no inflammatory response, but we did observe a subtle lesion involving tissue oedema and oedematous separation of epithelial tissues from underlying stroma. Oedema or a watery appearance is commonly reported in published descriptions of diseased molluscs, yet in many cases the terminology has been poorly characterised. The potential causes of oedema are reviewed; however, the question remains as to what might be the cause of oedema in molluscs that are normally iso-osmotic with seawater and have no power of anisosmotic extracellular osmotic regulation.

  18. Biogas production from anaerobic digestion of Spirulina maxima algal biomass

    SciTech Connect

    Samson, R.; LeDuy, A.

    1982-08-01

    The photosynthetic spectrum of solar energy could be exploited for the production of chemical energy of methane through the combined algal-bacterial process. In this process, the algae are mass produced from light and from carbon in the first step. The algal biomass is then used as a nutrient for feeding the anaerobic digester, in the second step, for the production of methane by anaerobic bacteria. The carbon source for the production of algal biomass could be either organic carbon from wastewaters (for eucaryotic algae), or carbon dioxide from the atmosphere or from the combustion exhaust gases (for both prokaryotic and eukaryotic algae). The technical feasibility data on the anaerobic digestion of algal biomass have been reported for many species of algae including macroscopic algae and microscopic algae. Research being conducted in the authors' laboratory consists of using the semimicroscopic blue-green alga Spirulina maxima as the sole substrate for this combined algal-bacterial process. This species of alga is very attractive for the process because of its capability of using the atmospheric carbon dioxide as carbon source and its simple harvesting methods. Furthermore, it appeared that the fermentability of S. maxima is significantly higher than other microscopic algae. This communication presents the results on the anaerobic inoculum development by the adaptation technique. This inoculum was then used for the semicontinuous anaerobic digestion of S. maxima algal biomass. The evolutions of biogas production and composition, biogas yield, total volatile fatty acids, alkalinity, ammonia nitrogen, pH, and electrode potential were followed.

  19. Interaction of Eimeria tenella with intestinal mucin in vitro.

    PubMed

    Tierney, J B; Matthews, E; Carrington, S D; Mulcahy, G

    2007-06-01

    The mucus gel layer overlying the gastrointestinal epithelium plays an important role in host-pathogen interactions. The initial interaction between the coccidian parasite Eimeria tenella and host cells of the intestinal epithelium must occur across this mucus interface. In this study, we examined the relationship between E. tenella and avian mucin, in particular the effect of purified intestinal regional mucin on parasite adherence and invasion in vitro. Secreted mucin from the chicken duodenum and cecum was purified by density gradient centrifugation and gel chromatography. Parasite invasion studies were performed in the Madin-Darby bovine kidney cell model. Eimeria tenella adherence to chicken duodenal mucin was detected, whereas adherence to cecal or bovine mucin was not shown. Parasite invasion into epithelial cells was not influenced by bovine mucin, whereas chicken mucin purified from the duodenum and cecum significantly inhibited invasion. Inhibition of E. tenella invasion into cells by mucin from the duodenum was marginally greater than that of the cecum, but this was not significant. This study demonstrated E. tenella interaction with native chicken intestinal mucin, which in turn inhibited parasite invasion into epithelial cells in vitro.

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

    PubMed

    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.

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    PubMed

    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.

  3. Increase of Clostridium perfringens in association with Eimeria in haemorrhagic enteritis in Japanese beef cattle.

    PubMed

    Kirino, Y; Tanida, M; Hasunuma, H; Kato, T; Irie, T; Horii, Y; Nonaka, N

    2015-08-18

    A coprological survey with detailed clinical observation of naturally occurring haemorrhagic enteritis (HE) cases was conducted to understand the pathophysiology of HE by clarifying the infection status of Eimeria and enteropathogenic bacteria in cattle. Faecal samples from 55 cases of HE and 26 clinically normal animals were collected, and a quantitative examination of Eimeria and potential enteropathogenic bacteria was performed. The number of Eimeria species oocysts per gram of faeces (OPG) exceeded 10,000 in 69.1 per cent of HE cases with a maximum of 1,452,500 OPG and Eimeria zuernii was found to be overwhelmingly dominant. A significant increase in faecal coliform count was observed in HE cases compared with clinically normal animals. Among the animals shedding >10,000 OPG, 42.9 per cent showed a remarkable increase in Clostridium perfringens abundance (>10(4) CFU/g) in the faeces. In the cases with C. perfringens detected, its abundance was positively correlated with Eimeria OPG and high C. perfringens abundance was always accompanied by high Eimeria OPG. E. zuernii is likely to play a crucial role in massive multiplication of C. perfringens in HE in cattle.

  4. Epidemiology of Eimeria infections in an Austrian milking sheep flock and control with diclazuril.

    PubMed

    Platzer, B; Prosl, H; Cieslicki, M; Joachim, A

    2005-04-20

    In a flock of milking sheep from Styria (Austria), the dynamics of Eimeria oocyst excretion was monitored in the lambing season 2003-2004 and a treatment trial with 1 mg/kg diclazuril (single dose on day 0 of study) was conducted in two groups of early weaned lambs. Adult animals (n = 30 ewes, 30 yearlings) excreted oocysts of different species (weekly prevalences from week -7 ante partum to week 5 post-partum: ewes 20-60%, yearlings 38-73%) in low intensities (< or = 6000 oocyst per gram of faeces, highest values in weeks -1 and -4) without clinical signs. Ewes excreted significantly fewer oocysts than yearlings. Lambs in the first group (n = 32 treated animals, 32 controls) excreted oocysts from the seventh day of sampling (average age on day 0: 28 days), those in the consecutive group (n = 32 treated animals, 30 controls) already excreted oocysts on day 0 (average age: 34 days). Treatment resulted in significant reduction of the overall excretion rates on days 7-21 of study compared with untreated controls. Similarly, the excretion intensities were significantly reduced in the treated groups. Overall excretion rates of the pathogenic species (Eimeria ovinoidalis, Eimeria bakuensis, Eimeria weybridgensis/Eimeria crandallis, Eimeria ahsata) were significantly reduced after treatment. Soft faeces and anal soiling in the lambs were significantly reduced and weight gain was increased. A single treatment of lambs with diclazuril before or shortly after the onset of oocyst shedding was sufficient to control oocyst excretion and improve animal health.

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

    PubMed

    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.

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

    PubMed

    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.

  7. Genotyping by sequencing reveals the interspecific C. maxima / C. reticulata admixture along the genomes of modern citrus varieties of mandarins, tangors, tangelos, orangelos and grapefruits.

    PubMed

    Oueslati, Amel; Salhi-Hannachi, Amel; Luro, François; Vignes, Hélène; Mournet, Pierre; Ollitrault, Patrick

    2017-01-01

    The mandarin horticultural group is an important component of world citrus production for the fresh fruit market. This group formerly classified as C. reticulata is highly polymorphic and recent molecular studies have suggested that numerous cultivated mandarins were introgressed by C. maxima (the pummelos). C. maxima and C. reticulata are also the ancestors of sweet and sour oranges, grapefruit, and therefore of all the "small citrus" modern varieties (mandarins, tangors, tangelos) derived from sexual hybridization between these horticultural groups. Recently, NGS technologies have greatly modified how plant evolution and genomic structure are analyzed, moving from phylogenetics to phylogenomics. The objective of this work was to develop a workflow for phylogenomic inference from Genotyping By Sequencing (GBS) data and to analyze the interspecific admixture along the nine citrus chromosomes for horticultural groups and recent varieties resulting from the combination of the C. reticulata and C. maxima gene pools. A GBS library was established from 55 citrus varieties, using the ApekI restriction enzyme and selective PCR to improve the read depth. Diagnostic polymorphisms (DPs) of C. reticulata/C. maxima differentiation were identified and used to decipher the phylogenomic structure of the 55 varieties. The GBS approach was powerful and revealed 30,289 SNPs and 8,794 Indels with 12.6% of missing data. 11,133 DPs were selected covering the nine chromosomes with a higher density in genic regions. GBS combined with the detection of DPs was powerful for deciphering the "phylogenomic karyotypes" of cultivars derived from admixture of the two ancestral species after a limited number of interspecific recombinations. All the mandarins, mandarin hybrids, tangelos and tangors analyzed displayed introgression of C. maxima in different parts of the genome. C. reticulata/C. maxima admixture should be a major component of the high phenotypic variability of this germplasm opening

  8. Computational prediction of absorbance maxima for a structurally diverse series of engineered green fluorescent protein chromophores.

    PubMed

    Timerghazin, Qadir K; Carlson, Haley J; Liang, Chen; Campbell, Robert E; Brown, Alex

    2008-02-28

    By virtue of its self-sufficiency to form a visible wavelength chromophore within the confines of its tertiary structure, the Aequorea victoria green fluorescent protein (GFP) is single-handedly responsible for the ever-growing popularity of fluorescence imaging of recombinant fusion proteins in biological research. Engineered variants of GFP with altered excitation or emission wavelength maxima have helped to expand the range of applications of GFP. The engineering of the GFP variants is usually done empirically by genetic modifications of the chromophore structure and/or its environment in order to find variants with new photophysical properties. The process of identifying improved variants could be greatly facilitated if augmented or guided by computational studies of the chromophore ground and excited-state properties and dynamics. In pursuit of this goal, we now report a thorough investigation of computational methods for prediction of the absorbance maxima for an experimentally validated series of engineered GFP chromophore analogues. The experimental dataset is composed of absorption maxima for 10 chemically distinct GFP chromophore analogues, including a previously unreported Y66D variant, measured under identical denaturing conditions. For each chromophore analogue, excitation energies and oscillator strengths were calculated using configuration interaction with single excitations (CIS), CIS with perturbative correction for double substitutions [CIS(D)], and time-dependent density functional theory (TD DFT) using several density functionals with solvent effects included using a polarizable continuum model. Comparison of the experimental and computational results show generally poor quantitative agreement with all methods attempted. However, good linear correlations between the calculated and experimental excitation energies (R2>0.9) could be obtained. Oscillator strengths obtained with TD DFT using pure density functionals also correlate well with the

  9. Norovirus recombination.

    PubMed

    Bull, Rowena A; Tanaka, Mark M; White, Peter A

    2007-12-01

    RNA recombination is a significant driving force in viral evolution. Increased awareness of recombination within the genus Norovirus of the family Calicivirus has led to a rise in the identification of norovirus (NoV) recombinants and they are now reported at high frequency. Currently, there is no classification system for recombinant NoVs and a widely accepted recombinant genotyping system is still needed. Consequently, there is duplication in reporting of novel recombinants. This has led to difficulties in defining the number and types of recombinants in circulation. In this study, 120 NoV nucleotide sequences were compiled from the current GenBank database and published literature. NoV recombinants and their recombination breakpoints were identified using three methods: phylogenetic analysis, SimPlot analysis and the maximum chi2 method. A total of 20 NoV recombinant types were identified in circulation worldwide. The recombination point is the ORF1/2 overlap in all isolates except one, which demonstrated a double recombination event within the polymerase region.

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

    USDA-ARS?s Scientific Manuscript database

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

  11. COMPARISON OF TRANSCRIPTIONAL RESONSES FROM AVIAN GUT TISSUES AFTER EIMERIA ACERVULINA AND E. MAXIMA INFECTIONS USING cDNA MICROARRAY TECHNOLOGY

    USDA-ARS?s Scientific Manuscript database

    Understanding the host response during pathogen infection will extend our knowledge of pathogenesis and enhance the development of novel preventive methodologies against important infectious diseases. Microarray technology is a powerful tool to analyze host transcriptional responses. Coccidiosis re...

  12. Iron ions increase the thermostability of phycocyanin of Spirulina maxima

    NASA Astrophysics Data System (ADS)

    Li, Jian-Hong; Tai, Zi-Hou; Tseng, Chao-Tsi

    1998-03-01

    A spectral method to investigate the effect of Fe3+, Fe2+ on the thermostability of phycocyanin (PC) of Spirulina maxima showed that iron ions provent decrease of visible light absorbance and fluorescence intensity of PC. Increase in denaturation temperature caused by Fe3+ was observed by the micro-differential scanning calorimetric method. All results showed iron ions maintain the aggregation stability of the PC. The absorption spectrum of phycocyanobilin (PCB, a prosthetic group of PC) with Fe3+ in chloroform was quite different from that of free PCB.

  13. Modeling weekly maxima PM10 concentration in Malaysia

    NASA Astrophysics Data System (ADS)

    Hasan, Husna; Mansor, Nadiah

    2014-09-01

    Weekly maxima of daily maximum PM10 concentration measures from the Air Pollutant Index (API) for thirteen monitoring stations in Malaysia are modeled using the Generalized Extreme Value (GEV) distribution. The L-moments method was used to estimate the parameter and the type of GEV distribution was determined from the 95% confidence intervals of shape parameter, ξ. The result from the Root Mean Square Error (RMSE) and the Mean Absolute Error (MAE) show that Alor Setar and Kemaman were the best stations that converged to the distribution. The return levels of PM10 concentration that is expected to exceed the maximum once within a selected period are obtained for all stations.

  14. Biogas production from anaerobic digestion of Spirulina maxima algal biomass

    SciTech Connect

    Rejean Samson; Anh LeDuy

    1982-08-01

    Spirulina maxima algal biomass could be used as the sole nutrient for the production of biogas by anaerobic digestion process. It is relatively simple to adapt the municipal sewage sludge to this new substrate. The adapted sludge is very stable. Under nonoptimal conditions, the methane yield and productivity obtained were 0.26 m/sup 3//(kg VS added day) and 0.26 m/sup 3//(kg VS added day), respectively, with the semicontinuous, daily fed, anaerobic digestion having loading rate of 0.97 kg VS/(m/sup 3/ day), retention time of 33 days and temperature of 30/sup 0/C.

  15. Two daily smoke maxima in eighteenth century London air

    NASA Astrophysics Data System (ADS)

    Harrison, R. Giles

    Varied electrostatics experiments followed Benjamin Franklin's pioneering atmospheric investigations. In Knightsbridge, Central London, John Read (1726-1814) installed a sensing rod in the upper part of his house and, using a pith ball electrometer and Franklin chimes, monitored atmospheric electricity from 1789 to 1791. Atmospheric electricity is sensitive to weather and smoke pollution. In calm weather conditions, Read observed two daily electrification maxima in moderate weather, around 9 am and 7 pm. This is likely to represent a double diurnal cycle in urban smoke. Before the motor car and steam railways, one source of the double maximum smoke pattern was the daily routine of fire lighting for domestic heating.

  16. Characterization of the antigen SO7 during development of Eimeria tenella.

    PubMed

    Fetterer, R H; Jenkins, M C; Miska, K B; Barfield, R C

    2007-10-01

    The developmental expression of the antigen SO7, which has been previously shown to protect chickens against infection by several Eimeria species, was investigated. Using RT-PCR, mRNA for SO7 was found to be restricted primarily to unsporulated oocysts (0 hr). Western blot (WB) analysis with an antibody to recombinant SO7 (rbSO7) revealed expression of the protein from 6 to 72 hr (fully sporulated) of sporulation and in sporozoites (SZ). SO7 was absent in host-derived second-stage merozoites (MZ) and was present in culture-derived first-stage MZ but at a level of only 25% of that exhibited by SZ. During invasion of Madin-Darby bovine kidney (MDBK) cells by SZ in vitro, the level of SO7 within cells, as determined by WB analysis, remained relatively constant until 48 hr of development and then decreased by about 40% at the next time point (72 hr). The SO7 secreted into the culture media during in vitro development increased to a relative maximum at 48 hr and then decreased to about 20% of maximum at 72 hr. Immunostaining with anti-rbSO7 indicates that SO7 is highly concentrated in both refractile bodies (RB) of SZ, with some limited distribution in the apical complex. Anti-rbSO7 intensively stained the intracellular parasites and the first-stage schizonts during in vitro development of E. tenella in MDBK cells. Upon release from the schizonts, the first-stage merozoites stained with 1 or 2 bright spots typically at each end. The results suggest that SO7 is closely associated with the SZ RB and is developmentally regulated but may not play a direct role in cellular invasion.

  17. Thermal profile of rabbits infected with Eimeria intestinalis.

    PubMed

    Vadlejch, Jaroslav; Knízková, Ivana; Makovcová, Katerina; Kunc, Petr; Jankovská, Ivana; Janda, Karel; Borkovcová, Marie; Langrová, Iva

    2010-08-04

    In this study, infrared thermography (IRT) was assessed as a means of detecting the changes in body temperature in rabbit coccidiosis, as well as determining IRT used in imaging and measuring the regional changes in skin temperature that occur in the rabbits during this infection. Rabbits were inoculated with 5 x 10(4) or 1 x 10(5) oocysts of Eimeria intestinalis, respectively. The group of control animals remained non-inoculated throughout the study. All measured temperatures (rectal, ocular surface and auricle pavilion temperatures) dropped significantly (P<0.05) in rabbits infected with 1 x 10(5) oocysts. This group of rabbits also showed a medium negative correlation in rectal temperature (r=-0.49), ocular surface temperature (r=-0.49) and auricle pavilion temperature (r=-0.49) and a small negative correlation in live body weight gain (r=-0.22) depending on the number of days elapsed from the time of infection exposure.

  18. In vitro efficacy of allicin on chicken Eimeria tenella sporozoites.

    PubMed

    Alnassan, Alaa Aldin; Thabet, Ahmed; Daugschies, Arwid; Bangoura, Berit

    2015-10-01

    Chicken coccidiosis is a major parasitic disease caused by Eimeria spp. It is controlled and treated using chemical anticoccidial agents. Development of partial or complete resistance toward these anticoccidials is considered a major problem in poultry industry. Allicin is an organosulfur compound produced as a result of the reaction between alliin and alliinase after hacking of garlic. In this study, tenfold dilution from 180 mg/ml to 1.8 ng/ml of allicin in distilled water was tested against E. tenella in vitro. The percent of inhibition in allicin was from 99.9 to 71.53% using 180 mg/ml and 180 ng/ml, respectively. The percent of inhibition was 56.24% using 1.8 ng/ml. We used allicin as a treatment from plants against chicken coccidiosis; however, in vivo study should be performed to confirm these results.

  19. Identification and characterization of secreted proteins in Eimeria tenella

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed

    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

  1. Monocyte- and macrophage-mediated immune reactions against Eimeria bovis.

    PubMed

    Taubert, Anja; Behrendt, Jan Hillern; Sühwold, Anke; Zahner, Horst; Hermosilla, Carlos

    2009-10-14

    Innate immune reactions conducted by macrophages may affect the outcome of primary infections and are crucial for the transition to adaptive immune responses. In bovine coccidiosis little is known on early monocyte/macrophage-mediated responses. We therefore investigated in vivo, in vitro and ex vivo reactions of monocytes and macrophages against Eimeria bovis, one of the most pathogenic Eimeria species in cattle. Macrophages significantly infiltrate the gut mucosa of E. bovis-infected calves, particularly after challenge infection. Furthermore, peripheral monocytes of infected animals, as precursor cells of macrophages, exhibited enhanced ex vivo phagocytic and oxidative burst activities. Enhanced levels of both activities were found early after infection and towards the end of first merogony. In vitro exposure of macrophages to sporozoites led to phagocytosis of the pathogen, whilst monocytes failed to do so. Phagocytosis occurred independently of the viability of the sporozoites, indicating that active invasion by the parasites was negligible. Phagocytosis occurred in the absence of immune serum, but could clearly be enhanced by addition of immune serum, suggesting macrophage-derived antibody-dependent cytotoxicity. Furthermore, co-culture of macrophages with sporozoites and stimulation with merozoite I antigen induced distinct levels of cytokine and chemokine gene transcription. Thus, the transcription of genes encoding for IFN-gamma, IL-12, TNF-alpha, IL-6, CXCL1, CXCL8, CXCL10 and COX-2 was upregulated after sporozoite encounter. In contrast, soluble merozoite I antigen only induced the gene transcription of IL-6 and IL-12 and failed to upregulate IFN-gamma and TNF-alpha gene transcripts. In monocytes, IFN-gamma and CXCL10 were found upregulated, all other immunoregulatory molecules tested were not affected. In summary, our results strongly suggest that macrophage-mediated, innate immune reactions play an important role in the early immune response to E

  2. Serine protease activity in developmental stages of Eimeria tenella.

    PubMed

    Fetterer, R H; Miska, K B; Lillehoj, H; Barfield, R C

    2007-04-01

    A number of complex processes are involved in Eimeria spp. survival, including control of sporulation, intracellular invasion, evasion of host immune responses, successful reproduction, and nutrition. Proteases have been implicated in many of these processes, but the occurrence and functions of serine proteases have not been characterized. Bioinformatic analysis suggests that the Eimeria tenella genome contains several serine proteases that lack homology to trypsin. Using RT-PCR, a gene encoding a subtilisin-like and a rhomboid protease-like serine protease was shown to be developmentally regulated, both being poorly expressed in sporozoites (SZ) and merozoites (MZ). Casein substrate gel electrophoresis of oocyst extracts during sporulation demonstrated bands of proteolytic activity with relative molecular weights (Mr) of 18, 25, and 45 kDa that were eliminated by coincubation with serine protease inhibitors. A protease with Mr of 25 kDa was purified from extracts of unsporulated oocysts by a combination of affinity and anion exchange chromatography. Extracts of SZ contained only a single band of inhibitor-sensitive proteolytic activity at 25 kDa, while the pattern of proteases from extracts of MZ was similar to that of oocysts except for the occurrence of a 90 kDa protease, resistant to protease inhibitors. Excretory-secretory products (ESP) from MZ contained AEBSF (4-[2-Aminoethyl] benzenesulphonyl fluoride)-sensitive protease activity with a specific activity about 10 times greater than that observed in MZ extracts. No protease activity was observed in the ESP from SZ. Pretreatment of SZ with AEBSF significantly reduced SZ invasion and the release of the microneme protein, MIC2. The current results suggest that serine proteases are present in all the developmental stages examined.

  3. Short Exon Detection via Wavelet Transform Modulus Maxima

    PubMed Central

    Zhang, Xiaolei; Shen, Zhiwei; Zhang, Guishan; Shen, Yuanyu; Chen, Miaomiao; Zhao, Jiaxiang; Wu, Renhua

    2016-01-01

    The detection of short exons is a challenging open problem in the field of bioinformatics. Due to the fact that the weakness of existing model-independent methods lies in their inability to reliably detect small exons, a model-independent method based on the singularity detection with wavelet transform modulus maxima has been developed for detecting short coding sequences (exons) in eukaryotic DNA sequences. In the analysis of our method, the local maxima can capture and characterize singularities of short exons, which helps to yield significant patterns that are rarely observed with the traditional methods. In order to get some information about singularities on the differences between the exon signal and the background noise, the noise level is estimated by filtering the genomic sequence through a notch filter. Meanwhile, a fast method based on a piecewise cubic Hermite interpolating polynomial is applied to reconstruct the wavelet coefficients for improving the computational efficiency. In addition, the output measure of a paired-numerical representation calculated in both forward and reverse directions is used to incorporate a useful DNA structural property. The performances of our approach and other techniques are evaluated on two benchmark data sets. Experimental results demonstrate that the proposed method outperforms all assessed model-independent methods for detecting short exons in terms of evaluation metrics. PMID:27635656

  4. Insight into the Salivary Transcriptome and Proteome of Dipetalogaster maxima

    PubMed Central

    Assumpção, Teresa C. F.; Charneau, Sébastien; Santiago, Paula B. M.; Francischetti, Ivo M. B.; Meng, Zhaojing; Araújo, Carla N.; Pham, Van M.; Queiroz, Rayner M. L.; de Castro, Cleudson Nery; Ricart, Carlos André; Santana, Jaime M.; Ribeiro, José M. C.

    2011-01-01

    Dipetalogaster maximais a blood-sucking Hemiptera that inhabits sylvatic areas in Mexico. It usually takes its blood meal from lizards, but following human population growth, it invaded suburban areas, feeding also on humans and domestic animals. Hematophagous insect salivary glands produce potent pharmacologic compounds that counteract host hemostasis, including anticlotting, antiplatelet, and vasodilatory molecules. To obtain further insight into the salivary biochemical and pharmacologic complexity of this insect, a cDNA library from its salivary glands was randomly sequenced. Salivary proteins were also submitted to one- and two-dimensional gel electrophoresis (1DE and 2DE) followed by mass spectrometry analysis. We present the analysis of a set of 2728 cDNA sequences, 1375 of which coded for proteins of a putative secretory nature. The saliva 2DE proteome displayed approximately 150 spots. The mass spectrometry analysis revealed mainly lipocalins, pallidipins, antigen 5-like proteins, and apyrases. The redundancy of sequence identification of saliva-secreted proteins suggests that proteins are present in multiple isoforms or derive from gene duplications. Supplemental files can be downloaded from http://exon.niaid.nih.gov/transcriptome/D_maxima/Dm-S1-web.xls and http://exon.niaid.nih.gov/transcriptome/D_maxima/Dm-S2-web.xls. PMID:21058630

  5. Can Spirulina maxima reduce the mutagenic potential of sibutramine?

    PubMed

    Araldi, R P; Santos, N P; Mendes, T B; Carvalho, L B; Ito, E T; de-Sá-Júnior, P L; Souza, E B

    2015-12-28

    The worldwide obesity pandemic requires the use of anti-obesity drugs. Sibutramine is an anti-obesity drug that has been used worldwide but is indiscriminately consumed in Brazil. Several studies have demonstrated that sibutramine promotes weight loss and weight maintenance, but several side effects have been associated with its systematic consumption. For this reason, sibutramine was withdrawn from the European and American markets, but still remains legal for use in Brazil. Studies have shown that a 5-10% reduction in body weight results in outstanding health benefits for obese patients. However, in order to promote significant weight loss, it is necessary to use sibutramine for at least 2 years. This long-term exposure has carcinogenic potential, as sibutramine causes DNA damage. Thus, this study evaluated the in vivo mutagenic potential of sibutramine alone (5, 7, 10, 15, and 20 mg/kg) and in association with Spirulina maxima (150 and 300 mg/kg), a cyanobacterium with antioxidant potential, using the polychromatic erythrocyte micronucleus test. Our results reinforced the mutagenic potential of sibutramine alone, which showed a time-dependent action. Combinatory treatments with S. maxima were not able to reduce the genotoxicity of sibutramine. These results were confirmed in vitro with the cytokinesis-blocked micronucleus test. In conclusion, our data showed that new alternative anti-obesity treatments are needed since the consumption of sibutramine can increase the risk of cancer in overweight patients.

  6. Expression of digestive enzymes and nutrient transporters in the small intestine of Eimeria acervulina-infected chickens

    USDA-ARS?s Scientific Manuscript database

    Coccidiosis is a major disease of poultry caused by the intestinal protozoa Eimeria. Eimeria acervulina mainly infects the duodenum, causing lesions in epithelial tissue. The objective of this study was to investigate the effect of E. acervulina infection on the expression of 18 nutrient transport...

  7. Cytokine gene expression and NF-kappaB activation following infection of intestinal epithelial cells with Eimeria bovis or Eimeria alabamensis in vitro.

    PubMed

    Alcala-Canto, Y; Ibarra-Velarde, F

    2008-03-01

    In cattle, Eimeria produces clinical disease with different degrees of severity, depending on the dominant species. Eimeria bovis triggers severe intestinal damage, while E. alabamensis causes minimal damage. Cytokines and other factors derived from epithelial cells play important roles in inflammatory and immune responses in intestinal tissue. This study aimed to obtain a detailed view of IFN-gamma and IL-4 mRNA expression as well as of activation of NF-kappaB p50 and p65 subunits induced by E. bovis or E. alabamensis in intestinal epithelial cells by means of a RT-PCR assay and a NF-kappaB p50/p65 ELISA-based kit, respectively. Our results demonstrate that infection by both Eimeria species enhances IL-4 mRNA expression in intestinal cells. However, IL-4 was expressed more intensely in cells incubated with E. bovis whereas IFN-gamma levels were higher and detected at an earlier time in cells infected with E. alabamensis. NF-kappaB was activated in infected cells irrespective of species, yet the activity of the p50 subunit was significantly higher in cells incubated with E. bovis. Our results suggest that the intensity of host-cell responses triggered by these two Eimeria species could be considered as potential determinants of pathogenicity.

  8. Cross-transmission studies with Eimeria arizonensis, E. arizonensis-like oocysts and Eimeria langebarteli: host specificity at the genus and species level within the Muridae.

    PubMed

    Hnida, J A; Duszynski, D W

    1999-10-01

    Cross-transmission experiments were done using sporulated oocysts of Eimeria arizonensis from Peromyscus truei and Peromyscus maniculatus, and oocysts of 2 putative species that resemble E. arizonensis, i.e., Eimeria albigulae from Neotoma albigula, and Eimeria onychomysis from Onychomys leucogaster. Oocysts of each species were inoculated into representatives of P. maniculatus and the latter 2 rodent species. Other experiments were conducted wherein oocysts of Eimeria langebarteli from Peromyscus leucopus were given to P. truei and P. maniculatus. Oocysts of E. arizonensis from P. truei and P. maniculatus could be transmitted only to P. maniculatus; likewise, oocysts of E. albigulae and E. onychomysis produced patent infections only in N. albigula and O. leucogaster, respectively. Oocysts of E. langebarteli from P. leucopus could be transmitted to P. truei, but not P. maniculatus. These results indicate that E. arizonensis, and the morphologically similar E. albigulae and E. onychomysis, are distinct species that are not transmissible between the genera of their respective hosts (Peromyscus, Neotoma, Onychomys), and that some isolates of E. langebarteli, reported from 6 species of Peromyscus and Reithrodontomys megalotis, may not always be infective to P. maniculatus.

  9. WITHDRAWN: Inhibition by toltrazuril of the recombinant cyclophilin ETCYP21 of Eimeria tenella.

    PubMed

    Bierbaum, Stefan; Mackiewicz, Monika; Wunderlich, Frank

    2010-10-12

    This article has been withdrawn at the request of the author(s) and/or editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy. Copyright © 2010. Published by Elsevier B.V. All rights reserved.

  10. Eimeria infection in camels (Camelus dromedarius) in Yazd province, central Iran.

    PubMed

    Sazmand, A; Hamidinejat, H; Hekmatimoghaddam, S; Asadollahi, Z; Mirabdollahi, S

    2012-03-01

    This study was carried out during the winter of 2008 to the summer 2010 to determine the rate of infection of Eimeria spp. in dromedary camels in Yazd province, Iran at that period. A total of 305 faecal samples were taken per recta from live and slaughtered, apparently healthy camels 9 months to 23 years old. Using saturated zinc sulfate solution floatation technique, samples were prepared and investigated microscopically to detect Eimeria spp. oocysts. Results revealed that the overall frequency of infection in samples was 9.51%. Identified species were Eimeria cameli (47.5%), Eimeria dromedarii (42.5%) and Eimeria bactriani (10%). The rate of infection was higher in the winter season, and in camels aged 5 to 10 years old. Statistical analysis showed that there is a significant difference between infection rate and season, but no effect by age or sex on eimeriosis was found. Since most of the positive cases in our study were adult, our findings suggest that older camels may play an important role in spreading infection as asymptomatic oocyst shedders.

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

    PubMed

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

    2016-05-01

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

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

    PubMed

    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.

  13. Montanide IMS 1313 N VG PR nanoparticle adjuvant enhances antigen-specific immune responses to profilin following mucosal vaccination against Eimeria acervulina.

    PubMed

    Jang, Seung I; Lillehoj, Hyun S; Lee, Sung Hyen; Lee, Kyung Woo; Lillehoj, Erik P; Bertrand, François; Dupuis, Laurent; Deville, Sébastien

    2011-12-15

    This study investigated protection against Eimeria acervulina (E. acervulina) following vaccination of chickens with an Eimeria recombinant profilin in conjunction with different adjuvants, or by changing the route of administration of the adjuvants. Day-old broilers were immunized twice with profilin emulsified in Montanide IMS 1313 N VG PR adjuvant (oral, nasal, or ocular routes), Montanide ISA 71 VG adjuvant (subcutaneous route), or Freund's adjuvant (subcutaneous route) and orally challenged with virulent E. acervulina parasites. Birds orally immunized with profilin plus IMS 1313 N VG PR, or subcutaneously immunized with profilin plus ISA 71 VG, had increased body weight gains compared with animals nasally or ocularly immunized with profilin plus IMS 1313 N VG PR, or subcutaneously immunized with profilin plus Freund's adjuvant. All adjuvant formulations, except for IMS 1313 N VG PR given by the nasal or ocular routes, decreased fecal parasite excretion and/or reduced intestinal lesions, compared with non-vaccinated and infected controls. Compared with animals vaccinated with profilin plus Freund's adjuvant, chickens immunized with profilin plus IMS 1313 N VG PR or ISA 71 VG showed higher post-infection intestinal levels of profilin-reactive IgY and secretary IgA antibodies. Finally, immunization with profilin in combination with ISA 71 VG was consistently better than profilin plus IMS 1313 N VG PR or Freund's adjuvant for increasing the percentages of CD4(+), CD8(+), BU1(+), TCR1(+), and TCR2(+) intestinal lymphocytes. These results indicate that experimental immunization of chickens with the recombinant profilin subunit vaccine in conjunction with IMS 1313 or ISA 71 VG adjuvants increases protective mucosal immunity against E. acervulina infection. Published by Elsevier B.V.

  14. Three new triterpene esters from pumpkin (Cucurbita maxima) seeds.

    PubMed

    Kikuchi, Takashi; Ueda, Shinsuke; Kanazawa, Jokaku; Naoe, Hiroki; Yamada, Takeshi; Tanaka, Reiko

    2014-04-16

    Three new multiflorane-type triterpene esters, i.e. 7α-hydroxymultiflor-8-ene-3α,29-diol 3-acetate-29-benzoate (1), 7α-methoxymultiflor-8-ene-3α,29-diol 3,29-dibenzoate (2), and 7β-methoxymultiflor-8-ene-3α,29-diol 3,29-dibenzoate (3), were isolated from seeds of Cucurbita maxima, along with the known compound, multiflora-7,9(11)-diene-3α,29-diol 3,29-dibenzoate (4). Compound 1 exhibited melanogenesis inhibitory activities comparable with those of arbutin. In cytotoxicity assays, compounds 1 and 3 exhibited weak cytotoxicity, with IC50 values of 34.5-93.7 μM against HL-60 and P388 cells.

  15. A microstructural study of individual nacre tablet of Pinctada maxima.

    PubMed

    Wang, Sheng-Nan; Yan, Xiao-Hui; Wang, Rizhi; Yu, Da-Hui; Wang, Xiao-Xiang

    2013-09-01

    Nacre tablets from the shell of Pinctada maxima were studied with SEM, TEM and STEM. The systematic nanolath morphology on the (001) surface of nacre tablets was observed after acidic etching and mechanical polishing. The nanolaths were along the [100] crystallographic orientation of aragonite crystal. The (010) and (100) cross section surfaces of the nacre tablets showed nanolath and nanograin morphologies, respectively, which was consistent with [100] crystallographic orientation of nanolath on the (001) surface. Sheet-like defects with low mass density were observed on the (001) plane inside nacre tablets and were considered to be the cause of nanolath morphology revealed on the surfaces by acidic etching and mechanical polishing. On the other hand, large block [110] twins that divide the nacre tablets into two sectors were identified. The implication of these twins on the understanding to the crystallization mechanism of nacre tablets was discussed.

  16. Maxima of two random walks: Universal statistics of lead changes

    SciTech Connect

    Ben-Naim, E.; Krapivsky, P. L.; Randon-Furling, J.

    2016-04-18

    In this study, we investigate statistics of lead changes of the maxima of two discrete-time random walks in one dimension. We show that the average number of lead changes grows as ${\\pi }^{-1}\\mathrm{ln}t$ in the long-time limit. We present theoretical and numerical evidence that this asymptotic behavior is universal. Specifically, this behavior is independent of the jump distribution: the same asymptotic underlies standard Brownian motion and symmetric Lévy flights. We also show that the probability to have at most n lead changes behaves as ${t}^{-1/4}{(\\mathrm{ln}t)}^{n}$ for Brownian motion and as ${t}^{-\\beta (\\mu )}{(\\mathrm{ln}t)}^{n}$ for symmetric Lévy flights with index μ. The decay exponent $\\beta \\equiv \\beta (\\mu )$ varies continuously with the Lévy index when $0\\lt \\mu \\lt 2$, and remains constant $\\beta =1/4$ for $\\mu \\gt 2$.

  17. Specific lignin accumulation in granulated juice sacs of Citrus maxima.

    PubMed

    Wu, Jia-Ling; Pan, Teng-Fei; Guo, Zhi-Xiong; Pan, Dong-Ming

    2014-12-17

    Juice sac granulation occurring in pummelo fruits [Citrus maxima (Burm.) Merr.] is an undesirable trait, and the underlying mechanism remains unresolved. Previous studies have shown that lignin metabolism is closely associated with the process of juice sac granulation. Here, a method suitable for lignin isolation from pummelo tissues is established. Acetylated lignins from different pummelo tissues and cultivars were analyzed by HSQC NMR. The results showed that lignins in granulated juice sacs were characterized by an extremely high abundance of guaiacyl units (91.13-96.82%), in contrast to lignins from other tissues, including leaves, stems, and segment membranes. The abnormally accumulated lignins in granulated juice sacs were specific and mainly polymerized from coniferyl alcohol. No significant difference was found in lignin types among various cultivars. These findings indicated that the mechanism of juice sac granulation might be similar among various cultivars, although very different degrees of juice sac granulation can be observed.

  18. Parameter Space of the Columbia River Estuarine Turbidity Maxima

    NASA Astrophysics Data System (ADS)

    McNeil, C. L.; Shcherbina, A.; Lopez, J.; Karna, T.; Baptista, A. M.; Crump, B. C.; Sanford, T. B.

    2016-12-01

    We present observations of estuarine turbidity maxima (ETM) in the North Channel of the Columbia River estuary (OR and WA, USA) covering different river discharge and flood tide conditions. Measurements were made using optical backscattering sensors on two REMUS-100 autonomous underwater vehicles (AUVs) during spring 2012, summer 2013, and fall 2012. Although significant short term variability in AUV measured optical backscatter was observed, some clustering of the data occurs around the estuarine regimes defined by a mixing parameter and a freshwater Froude number (Geyer & MacCready [2014]). Similar clustering is observed in long term time series of turbidity from the SATURN observatory. We will use available measurements and numerical model simulations of suspended sediment to further explore the variability of suspended sediment dynamics within a frame work of estuarine parameter space.

  19. Maxima of two random walks: Universal statistics of lead changes

    SciTech Connect

    Ben-Naim, E.; Krapivsky, P. L.; Randon-Furling, J.

    2016-04-18

    In this study, we investigate statistics of lead changes of the maxima of two discrete-time random walks in one dimension. We show that the average number of lead changes grows as ${\\pi }^{-1}\\mathrm{ln}t$ in the long-time limit. We present theoretical and numerical evidence that this asymptotic behavior is universal. Specifically, this behavior is independent of the jump distribution: the same asymptotic underlies standard Brownian motion and symmetric Lévy flights. We also show that the probability to have at most n lead changes behaves as ${t}^{-1/4}{(\\mathrm{ln}t)}^{n}$ for Brownian motion and as ${t}^{-\\beta (\\mu )}{(\\mathrm{ln}t)}^{n}$ for symmetric Lévy flights with index μ. The decay exponent $\\beta \\equiv \\beta (\\mu )$ varies continuously with the Lévy index when $0\\lt \\mu \\lt 2$, and remains constant $\\beta =1/4$ for $\\mu \\gt 2$.

  20. Acylated quercetagetin glycosides with antioxidant activity from Tagetes maxima.

    PubMed

    Parejo, Irene; Bastida, Jaume; Viladomat, Francesc; Codina, Carles

    2005-10-01

    The fractionation of a methanolic extract of Tagetes maxima guided for antioxidant activity resulted in the isolation of three acylated quercetagetin glycosides, quercetagetin-7-O-(6-O-caffeoyl-beta-D-glucopyranoside), quercetagetin-7-O-(6-O-p-coumaroyl-beta-D-glucopyranoside) and quercetagetin-7-O-(6-O-galloyl-beta-D-glucopyranoside), as well as four known flavonoid glycosides. The structural elucidation was accomplished by spectroscopic methods (ESI-MS/MS and NMR). The antioxidant activity of fractions and isolated compounds was determined by checking the scavenging activity against three different radicals: 2,2-diphenyl-1-picrylhydrazyl free radical (DPPH*), hydroxyl (*OH), and superoxide (O2*-). The three isolated compounds exhibited a high radical scavenging activity in comparison with reference compounds.

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

    PubMed

    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.

  2. Genetic Recombination

    ERIC Educational Resources Information Center

    Whitehouse, H. L. K.

    1973-01-01

    Discusses the mechanisms of genetic recombination with particular emphasis on the study of the fungus Sordaria brevicollis. The study of recombination is facilitated by the use of mutants of this fungus in which the color of the ascospores is affected. (JR)

  3. Genetic Recombination

    ERIC Educational Resources Information Center

    Whitehouse, H. L. K.

    1973-01-01

    Discusses the mechanisms of genetic recombination with particular emphasis on the study of the fungus Sordaria brevicollis. The study of recombination is facilitated by the use of mutants of this fungus in which the color of the ascospores is affected. (JR)

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

    PubMed

    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.

  5. Prevalence of Eimeria spp., Cryptosporidium spp. and Giardia spp. in calves in the Van province.

    PubMed

    Gül, Abdurrahman; Ciçek, Mutalip; Kilinç, Ozlem

    2008-01-01

    This research was carried out in order to determine the prevalence of Eimeria spp. Cryptosporidium spp. oocysts and Giardia cysts in calves less than 6 months of age in Van province. For this purpose, fecal samples were obtained from the rectum of 182 calves. Fecal samples (n: 182) were examined with the modified acid-fast technique for Cryptosporidium spp. oocysts. The same samples were examined by zinc sulphate flotation technique for Eimeria oocysts and Giardia cysts. During the laboratory examination of fecal samples, Eimeria spp. oocysts were identified in 22.53% (41/182), Cryptosporidium oocysts in 13.19% (24/182) and Giardia cysts in 9.34 % (17/182) of the dairy calves examined. The rate of infection was 69.78% (127/182). Single infections (45.05%) and mixed infections (24.73%) were identified.

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

    PubMed Central

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

    PubMed

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

    2009-12-23

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

  8. Prevalence of Eimeria infection in yaks on the Qinghai-Tibet Plateau of China.

    PubMed

    Dong, Hui; Li, Chunhua; Zhao, Qiping; Li, Jing; Han, Hongyu; Jiang, Lianlian; Zhu, Shunhai; Li, Ting; Kong, Chunlin; Huang, Bing; Cai, Jinzhong

    2012-10-01

    Few data are available on the prevalence of Eimeria spp. in yaks. An observational study was conducted to determine the prevalence of coccidial infection in yaks on the Qinghai-Tibet Plateau of China. A total of 324 fecal samples from 4 counties was examined, and oocysts were identified to the species level on the basis of morphological features. Eimeria oocysts were found in 113 (34.9%) samples. The species detected and their prevalence values included the following: Eimeria zuernii (54.9%), E. pellita (35.4%), E. canadensis (33.6%), E. bovis (23.0%), E. cylindrica (16.8%), E. subspherica (16.8%), E. ellipsoidalis (14.1%), E. brasiliensis (13.3%), E. wyomingensis (8.0%), E. alabamensis (7.1%), E. illinoisensis (5.3%), E. auburnensis (4.4%), E. bombayansis (3.5%), and E. bukidnonensis (2.7%). Mixed infections of 2 to 7 species were found in 66.4% of the animals. There was an age-related difference in the prevalence of infection. The highest prevalence (53.3%) was observed in calves, an intermediate prevalence in yearlings (36.1%), and the lowest was in adults (15.6%). The number of oocysts per g of feces was significantly higher in calves than in adults. More Eimeria species were indentified in calves. Eimeria zuernii was the most prevalent species in calves and adults, whereas in yearling yaks E. pellita was most common. The majority of calves and yearlings showed mixed infection, but adults tended to be infected with 1 species. The prevalence and intensity of Eimeria species were found to show statistically significant differences among different regions in Qinghai Province.

  9. Cell: sporozoite interactions and invasion by apicomplexan parasites of the genus Eimeria.

    PubMed

    Augustine, P C

    2001-01-01

    The site specificity that avian Eimeria sporozoites and, to a more limited degree, other apicomplexan parasites exhibit for invasion in vivo suggests that specific interactions between the sporozoites and the target host cells may mediate the invasion process. Although sporozoite motility and structural and secreted antigens appear to provide the mechanisms for propelling the sporozoite into the host cell,there is a growing body of evidence that the host cell provides characteristics by which the sporozoites recognise and interact with the host cell as a prelude to invasion. Molecules on the surface of cells in the intestinal epithelium, that act as receptor or recognition sites for sporozoite invasion, may be included among these characteristics. The existence of receptor molecules for invasion by apicomplexan parasites was suggested by in vitro studies in which parasite invasion was inhibited in cultured cells that were treated with a variety of substances designed to selectively alter the host cell membrane. These substance included cationic compounds or molecules, enzymes that cleave specific linkages, protease inhibitors, monoclonal antibodies, etc. More specific evidence for the presence of receptors was provided by the binding of parasite antigens to specific host cell surface molecules. Analyses of host cells have implicated 22, 31, and 37 kDa antigens, surface membrane glycoconjugates,conserved epitopes of host cells and sporozoites, etc., but no treatment that perturbs these putative receptors has completely inhibited invasion of the cells by parasites. Regardless of the mechanism,sporozoites of the avian Eimeria also invade the same specific sites in foreign host birds that they invade in the natural host. Thus, site specificity for invasion may be a response to characteristics of the intestine that are shared by a number of hosts rather than to a unique trait of the natural host. Protective immunity elicited against avian Eimeria species is not

  10. Coccidia of wombats: correction of host-parasite relationships. Eimeria wombati (Gilruth and Bull, 1912) Comb. Nov. and Eimeria ursini Supperer, 1957 from the hairy-nosed wombat and Eimeria arundeli sp. n. from the common wombat.

    PubMed

    Barker, I K; Munday, B L; Presidente, P J

    1979-06-01

    Coccidial oocysts morphologically consistent with Eimeria ursini Supperer 1957, and E. tasmaniae Supperer 1957 were recovered from the feces of wild and captive hairy-nosed wombats (Lasiorhinus latifrons) in Australia. Eimeria arundeli so. n. was recovered from the feces of wild and captive common wombats (Vombatus ursinus). Eimeria arundeli oocysts are ellipsoidal to slightly ovoid 60.2--67.2 (63.7) X 40.6--47.6 (43.4); micropyle 3 in diameter usually visible; with oocyst wall granular, dark brown and occasionally opaque, 4--7 thick; inner oocyst wall clear, about 1.5 thick; small oocyst residuum present, four sporocysts ovoid 22.4--29.4 (25.8) X 12.6--15.4 (14.1) with protuberant Stieda body; opposite end of sporocyst also often slighly pointed; large granular sporocyst residuum obscuring sporozoites. Gametocytes of E. arundeli sp. n. and of an organism which is consistent with E. tasmaniae, are described developing in the lamina propria of villi in the small intestine. The stages in the hairy-nosed wombat are those described as Ileocystis wombati Gilruth and Bull 1912. It is suggested that the identification of the host of Supperer's E. ursini and E. tasmaniae as V. ursinus was in error and that the allopatric L. latifrons is the natural host. Eimeria tasmaniae Supperer 1957 is suppressed and E. wombati (Gilruth and Bull, 1912) comb. nov. is proposed and redescribed. No schizonts were identified among the endogenous stages, consistent with observations in the literature on other coccidia with similar gametocyte and oocyst structure.

  11. Eimeria bovis: an update on parasite-host cell interactions.

    PubMed

    Hermosilla, Carlos; Ruiz, Antonio; Taubert, Anja

    2012-10-01

    Apicomplexan parasites are obligate intracellular protozoans and are well recognized modulators of the host cell machinery on varying levels such as host cell metabolism, MHC expression, cell cycle, or apoptosis in order to guarantee their intracellular development and survival. One of the most thoroughly examined apicomplexan pathogens demonstrating a potent manipulative capacity with respect to various host cell functions is Toxoplasma gondii, a protozoon exhibiting rapid intracellular development with small meronts in any nucleated cell, almost irrespective of the cell type or host origin. In contrast, Eimeria bovis merogony I is host- and cell type-restricted and occurs exclusively in bovine endothelial host cells. Furthermore, as a peculiarity, intracellular E. bovis meront I development is a long-lasting process (up to 3 weeks), leading to the formation of huge macromeronts of up to 300 μm in size, containing up to 120,000 merozoites I as offspring. In consequence, the necessity for intense host cell modulation to support this particular development appears even more pressing than in other apicomplexan parasite cases. Here we review the data currently available on E. bovis-host cell interactions, indicating the intriguing capacity of this protozoan to exploit and utilize its host cell for its own benefit.

  12. PMN-mediated immune reactions against Eimeria bovis.

    PubMed

    Behrendt, Jan Hillern; Hermosilla, Carlos; Hardt, Martin; Failing, Klaus; Zahner, Horst; Taubert, Anja

    2008-02-14

    For successful in vivo infection, Eimeria bovis sporozoites have to traverse the mucosal layer of the ileum to infect lymphatic endothelial cells and may, thereby, be exposed to the interstitial fluid and to the lymph representing potential targets for leukocytes. To mimic this situation in vitro, we exposed E. bovis sporozoites to bovine PMN and found enhanced elimination of the parasites. Addition of immune serum clearly increased these reactions, whereas neonatal calf serum had no effect, thus proposing a PMN-derived antibody-dependent cytotoxicity. Scanning and transmission electron microscopy showed PMN engulfing sporozoites or extending filopodia towards them and occasionally incorporating the parasites. PMN reacted with enhanced transcription of IL-6, MCP-1, GROalpha, TNF-alpha, and iNOS genes after exposure to sporozoites while stimulation with merozoite-antigen, in addition, upregulated IL-8, IP-10 and IL-12 gene transcription. Furthermore, enhanced in vitro oxidative burst and phagocytic activities were observed after contact of PMN with viable sporozoites. To verify the potential role of PMN in the in vivo situation, we analysed the general phagocytic and oxidative burst activities of PMN obtained ex vivo from E. bovis experimentally infected calves. Enhanced levels of both activities were found early p.i. (1-5 days) and towards the end of the first schizogony (days 13-22 p.i.) underlining the in vitro data. Our results suggest that PMN-mediated, innate immune reactions play an important role in the early immune response to E. bovis infections in calves.

  13. Ultrastructural effects of acetamizuril on endogenous phases of Eimeria tenella.

    PubMed

    Liu, Lili; Chen, Huiya; Fei, Chenzhong; Wang, Xiaoyang; Zheng, Wenli; Wang, Mi; Zhang, Keyu; Zhang, Lifang; Li, Tao; Xue, Feiqun

    2016-03-01

    To explore the primary stage or site of action of acetamizuril (AZL), a novel triazine anticoccidial compound, the ultrastructural development of Eimeria tenella at different endogenous stages was studied in experimentally infected chickens treated with a single oral dose of 15 mg/kg AZL. As a result of drug action, the differentiations of second-generation schizonts and microgamonts were largely inhibited and merozoites became irregular in shape. Meanwhile, the outer membrane blistering and perinuclear space enlargement were obvious in the second-generation schizonts and microgamonts, which were never observed in the classic triazine anticoccidiosis drug diclazuril-treated E. tenella. The chromatin aggregation, anachromasis, and marginalization were visible in merozoites and microgamonts. Furthermore, the abnormal evagination of microgametes finally resulted in the degeneration of microgamonts and the failure of subsequent fertilization. The most marked micromorphological alteration occurring in the macrogamonts was the WFB2. Even if the fertilization occurred, the formation of oocyst wall became malformed and the zygote proceeded to the obvious degeneration. In addition, mitochondria swelling and cytoplasm vacuolization were discerned in respective intracellular stages, while endoplasmic reticulum and Golgi body swelling was less seen. These alterations may be the causes leading to the final death of E. tenella and also provide some information for further exploring the mechanism of action of AZL at the molecular level.

  14. Eimeria tenella: 14-3-3 protein interacts with telomerase.

    PubMed

    Zhao, Na; Gong, Pengtao; Cheng, Baiqi; Li, Jianhua; Yang, Zhengtao; Li, He; Yang, Ju; Zhang, Guocai; Zhang, Xichen

    2014-10-01

    Telomerase, consisting of telomerase RNA and telomerase reverse transcriptase (TERT), is responsible for the maintenance of the end of linear chromosomes. TERT, as the catalytic subunit of telomerase, plays a critical role in telomerase activity. Researches indicate TERT-associated proteins participate in the regulation of telomerase assembly, posttranslational modification, localization, and enzymatic function. Here, the telomerase RNA-binding domain of Eimeria tenella TERT (EtTRBD) was cloned into pGBKT7 and performed as the bait. α-Galactosidase assay showed that the bait plasmid did not activate Gal4 reporter gene. Further, we isolated an EtTRBD-associated protein, 14-3-3, by yeast two-hybrid screening using the constructed bait plasmid. To confirm the interaction, EtTRBD and 14-3-3 were expressed by prokaryotic and eukaryotic expression systems. Pull-down assays by purified proteins demonstrated a direct bind between EtTRBD and 14-3-3. Co-immunoprecipitation techniques successfully validated that 14-3-3 interacted with EtTRBD in 293T cells. The protein-protein interaction provides a starting point for more in-depth studies on telomerase and telomere regulation in E. tenella.

  15. Effects of curcumin (diferuloylmethane) on Eimeria tenella sporozoites in vitro.

    PubMed

    Khalafalla, Reda E; Müller, Uwe; Shahiduzzaman, Md; Dyachenko, Viktor; Desouky, Abdelrazik Y; Alber, Gottfried; Daugschies, Arwid

    2011-04-01

    The negative effects of coccidiosis on poultry health and productivity and increasing problems related to drug resistance have stimulated the search for novel and alternative methods of control. The present study evaluates the anticoccidial activity of curcumin (diferuloylmethane), a natural polyphenolic compound abundant in the rhizome of the perennial herb turmeric (Curcuma longa) which is a spice and food colorant commonly used in curries and also used as medicinal herb. Its effects were evaluated on Eimeria tenella sporozoites, including morphological alterations, sporozoite viability and infectivity to Madin-Darby bovine kidney (MDBK) cells. Morphological alterations of the sporozoites were recorded as deformation due to swelling and cell membrane corrugations. Curcumin at concentrations of 25, 50, 100, 200 and 400 μM showed considerable effects on sporozoite morphology and viability in a dose-dependent manner after incubation over 3, 6, 18 and 24 h while lower curcumin concentrations (6.25 and 12.5 μM) were not effective. In comparison to the untreated control, sporozoite infectivity was reduced at curcumin concentrations of 100 and 200 μM by 41.6% and 72.8%, respectively. Negative effects of curcumin on MDBK cells were not seen at these concentrations; however, curcumin at concentrations of 1,800, 600 and 400 μM was toxic to MDBK cells and affected cell proliferation. In conclusion, curcumin exhibited a marked inhibitory effect in vitro on E. tenella sporozoites inducing morphological changes and reducing sporozoite viability and infectivity.

  16. Partial purification and characterization of an aminopeptidase from Eimeria tenella.

    PubMed

    Fetterer, R H; Miska, K B; Barfield, R C

    2005-12-01

    Our previous investigation demonstrated the expression in Eimeria tenella sporulated oocysts of an aminopeptidase (AP) with strong homology to AP N. To further understand the role of proteases during development, we investigated the molecular and biochemical properties of E. tenella AP. Greater than 95% AP activity was present in a soluble extract during sporulation of oocysts with highest activity in fully sporulated oocysts. The AP activity was inhibited by the AP inhibitors bestatin and 1,6-phenanthroline, but not by serine protease inhibitors. The AP had specificity for synthetic endopeptidase substrates that contain arginine, alanine, or glycine at the N terminus. Partial purification of the enzyme yielded a major protein band with an Mr of about 106 kDa and an isoelectric point (Ip) of 5.1. Reverse transcription-polymerase chain reaction indicated that the gene for AP is expressed during sporulation, but expression is absent or greatly reduced in the sporozoites and merozoites. On the basis of the deduced gene structure, the predicted Mr is 110 kDa with a pI of 5.59. Database search indicates that the E. tenella AP shares significant homology with the AP from Apicomplexan taxa: Toxoplasma gondii, Cryptosporidium parvum, and Cryptosporidium hominis. Together, these results confirm the presence of a cytosolic AP related to AP N, which is expressed and active during sporulation of E. tenella oocysts.

  17. Molecular and biochemical characterization of Eimeria tenella hexokinase.

    PubMed

    Sun, Mingfei; Liao, Shenquan; Zhang, Longxian; Wu, Caiyan; Qi, Nanshan; Lv, Minna; Li, Juan; Lin, Xuhui; Zhang, Jianfei; Xie, Mingquan; Zhu, Guan; Cai, Jianping

    2016-09-01

    Hexokinase (HK) is one of the key enzymes in the glycolytic pathway that catalyzes the phosphorylation of glucose. In the present study, we cloned the HK gene from the coccidian Eimeria tenella (EtHK), expressed EtHK as a His-tagged fusion protein, and characterized its primary biochemical features. Mutagenesis confirmed that residues S159, N216, and D217 are essential or important to the EtHK catalytic activity. EtHK exhibited high affinity for D-glucose (Km = 0.67 to 0.79 mM), but was also able to utilize 2-deoxy-D-glucose (Km = 5.66 mM), D-fructose (Km = 13.76 mM), and D-mannose (Km = 25.41 mM). We also observed that quercetin and mangiferin could inhibit the EtHK enzyme activity (IC50 values = 6.52 and 85.82 μM, respectively). Among the two inhibitors, mangiferin also inhibited the growth of E. tenella in vitro (MIC50 = 0.12 μM). These observations suggest that EtHK may be explored as potential drug target, and mangiferin and its analogs may be explored for developing anti-coccidial therapeutics.

  18. Effect of betaine on the growth performance of chicks inoculated with mixed cultures of avian Eimeria species and on invasion and development of Eimeria tenella and Eimeria acervulina in vitro and in vivo.

    PubMed

    Augustine, P C; McNaughton, J L; Virtanen, E; Rosi, L

    1997-06-01

    At 7 d postinoculation (DPI) with a mixed culture of avian Eimeria species, 21-d-old chicks maintained in batteries and floor pens on a diet containing 0.15% (3 lb/ton) betaine plus 66 ppm (60 g/ton) salinomycin were significantly heavier and had significantly lower feed conversion ratios and mortality than chicks fed diets containing 0.15% betaine or 66 ppm salinomycin alone, or the control diet. At 31 DPI, when the chicks were 45 d old, the differences between the diet groups were not as great as at 7 DPI. In vitro, except at high concentrations, betaine was nontoxic to sporozoites of Eimeria tenella or Eimeria acervulina and had little effect on their invasion and development in cultured cells. In vivo, invasion by E. tenella and E. acervulina sporozoites was significantly reduced in all chicks fed diets containing betaine or salinomycin compared with that in control chicks. There was a significant interaction between betaine and salinomycin that impacted on invasion by both species. Overall development of E. tenella did not appear to be adversely affected by addition of betaine to diets containing salinomycin. Conversely, development of E. acervulina was reduced in chicks fed diets containing 0.075% (1.5 lb/ton) betaine plus 66 ppm salinomycin as compared with that in chicks fed salinomycin alone.

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

    PubMed

    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.

  20. VizieR Online Data Catalog: Minima and maxima of 299 variables (Hubscher+, 2009)

    NASA Astrophysics Data System (ADS)

    Hubscher, J.; Steinbach, H.-M.; Walter, F.

    2009-03-01

    In this 61th compilation of BAV results, photoelectric observations obtained in the year 2008 are presented on 299 variable stars giving 655 minima on eclipsing binaries and maxima on pulsating stars. All moments of minima and maxima are heliocentric. (3 data files).

  1. Critical thermal maxima of two geographic strains of channel and hybrid catfish

    USDA-ARS?s Scientific Manuscript database

    Critical thermal maxima have been used extensively to provide physiologically and ecologically valuable reference points that identify early signs of thermal stress. In catfish pond culture, daily temperature maxima up to 36'C and daily fluctuations of as much as 6'Care observed. These extreme condi...

  2. Cultivation of Spirulina maxima in medium supplemented with sugarcane vinasse.

    PubMed

    dos Santos, Raquel Rezende; Araújo, Ofélia de Queiroz Fernandes; de Medeiros, José Luiz; Chaloub, Ricardo Moreira

    2016-03-01

    The feasibility of sugarcane vinasse as supplement in growth medium of Spirulina maxima was investigated. The cell was cultivated under autotrophic (no vinasse, 70 μmol photons m(-2) s(-1)), heterotrophic (no light, culture medium supplemented with vinasse at 0.1% v/v and 1.0% v/v) and mixotrophic conditions (70 μmol photons m(-2) s(-1), vinasse at 0.1% v/v and 1.0% v/v). These preliminary results suggested a cyclic two-stage cultivation - CTSC, with autotrophic condition during light phase of the photoperiod (12 h, 70-200 μmol photons m(-2) s(-1)) and heterotrophic condition during dark phase (12h, 3.0% v/v vinasse). The adopted CTSC strategy consisted in three cycles with 75% withdrawal of suspension and reposition of medium containing 3.0% v/v vinasse, separated by autotrophic rest periods of few days between cycles. Results show an increase of biomass concentration between 0.495 g L(-1) and 0.609 g L(-1) at the 7th day of each cycle and high protein content (between 74.3% and 77.3% w/w).

  3. Extremal Regions Detection Guided by Maxima of Gradient Magnitude.

    PubMed

    Faraji, Mehdi; Shanbehzadeh, Jamshid; Nasrollahi, Kamal; Moeslund, Thomas Baltzer

    2015-12-01

    A problem of computer vision applications is to detect regions of interest under different imaging conditions. The state-of-the-art maximally stable extremal regions (MSERs) detects affine covariant regions by applying all possible thresholds on the input image, and through three main steps including: (1) making a component tree of extremal regions' evolution; (2) obtaining region stability criterion; and (3) cleaning up. The MSER performs very well, but, it does not consider any information about the boundaries of the regions, which are important for detecting repeatable extremal regions. We have shown in this paper that employing prior information about boundaries of regions results in a novel region detector algorithm that not only outperforms MSER, but avoids the MSER's rather complicated steps of enumeration and the cleaning up. To employ the information about the region boundaries, we introduce maxima of gradient magnitudes (MGMs) which are shown to be points that are mostly around the boundaries of the regions. Having found the MGMs, the method obtains a global criterion for each level of the input image which is used to find extremum levels (ELs). The found ELs are then used to detect extremal regions. The proposed algorithm which is called extremal regions of extremum levels (EREL) has been tested on the public benchmark data set of Mikolajczyk. The obtained experimental results show that the inclusion of region boundaries through MGMs, results in a detector that detects regions with high repeatability scores and is more robust against noise compared with MSER.

  4. Maxima of two random walks: Universal statistics of lead changes

    DOE PAGES

    Ben-Naim, E.; Krapivsky, P. L.; Randon-Furling, J.

    2016-04-18

    In this study, we investigate statistics of lead changes of the maxima of two discrete-time random walks in one dimension. We show that the average number of lead changes grows asmore » $${\\pi }^{-1}\\mathrm{ln}t$$ in the long-time limit. We present theoretical and numerical evidence that this asymptotic behavior is universal. Specifically, this behavior is independent of the jump distribution: the same asymptotic underlies standard Brownian motion and symmetric Lévy flights. We also show that the probability to have at most n lead changes behaves as $${t}^{-1/4}{(\\mathrm{ln}t)}^{n}$$ for Brownian motion and as $${t}^{-\\beta (\\mu )}{(\\mathrm{ln}t)}^{n}$$ for symmetric Lévy flights with index μ. The decay exponent $$\\beta \\equiv \\beta (\\mu )$$ varies continuously with the Lévy index when $$0\\lt \\mu \\lt 2$$, and remains constant $$\\beta =1/4$$ for $$\\mu \\gt 2$$.« less

  5. The complete mitochondrial genome of Acanthastrea maxima (Cnidaria, Scleractinia, Lobophylliidae).

    PubMed

    Arrigoni, Roberto; Vacherie, Benoît; Benzoni, Francesca; Barbe, Valérie

    2016-01-01

    The complete nucleotide sequence of the mitochondrial genome of the scleractinian coral Acanthastrea maxima has been obtained, representing the first sequenced mitogenome of a member of the Lobophylliidae. The mitochondrial genome is 18,278  bp in length, the longest sequence among the robust corals sequenced mitogenome to date. The overall GC composition (33.7%) and the gene arrangement are similar to those of the other scleractinian corals, including 13 protein-coding genes, 2 rRNA genes (rnl and rns) and 2 tRNA genes (tRNA-Met and tRNA-Trp). All genes except tRNA-Trp, atp8, cox1, tRNA-Met and rnl are engulfed by a large group I intron in the nad5 gene. A second group I intron of 1077 bp in length is inserted in the cox1 gene and it encodes a putative homing endonuclease. There are four regions of gene overlaps totalling 22 bp and nine intergenic spacer regions for a total of 2220 bp, of which the cox3-cox2 region may correspond to the putative control region.

  6. Phenoloxidase activity of Helix aspersa maxima (garden snail, gastropod) hemocyanin.

    PubMed

    Raynova, Yuliana; Doumanova, Lyuba; Idakieva, Krassimira Nikolova

    2013-12-01

    The oxygen-transporting protein, hemocyanin (Hc), of the garden snail Helix aspersa maxima (HaH) was isolated and kinetically characterized. Kinetic parameters of the reaction of catalytic oxidation of catechol to quinone, catalyzed by native HaH were determined: the V max value amounted to 22 nmol min(-1) mg(-1), k cat to 1.1 min(-1). Data were compared to those reported for other molluscan Hcs and phenoloxidases (POs). The o-diphenoloxidase activity of the native HaH is about five times higher than the activity determined for the Hcs of the terrestrial snail Helix pomatia and of the marine snail Rapana thomasiana (k cat values of 0.22 and 0.25 min(-1), respectively). The K m values obtained for molluscan Hcs from different species are comparable to those for true POs, but the low catalytic efficiency of Hcs is probably related to inaccessibility of the active sites to potential substrates. Upon treatment of HaH with subtilisin DY, the enzyme activity against substrate catechol was considerably increased. The relatively high proteolytically induced o-diPO activity of HaH allowed using it for preparation of a biosensor for detection of catechol.

  7. Year-long presence of Eimeria echidnae and absence of Eimeria tachyglossi in captive short-beaked echidnas ( Tachyglossus aculeatus ).

    PubMed

    Debenham, John J; Johnson, Robert; Vogelnest, Larry; Phalen, David N; Whittington, Richard; Slapeta, Jan

    2012-06-01

    The short-beaked echidna ( Tachyglossus aculeatus ) is 1 of 5 extant species of monotreme, found only in Australia and Papua New Guinea. The aim of this study was to identify the species of coccidia present and establish a range of subclinical Eimeria spp. (Coccidia: Apicomplexa) oocyst shedding in echidnas from eastern Australia over 18 mo. The coccidia were detected in 89% (49/55) of fecal samples from 12 long-term monitored and healthy captive echidnas, 75% (3/4) of 4 healthy long-term captive echidnas, 83% (5/6) of 6 short-term captive echidnas, and 60% (6/10) of 10 wild echidnas. Echidnas captive for 4 to 23 yr shed 100-46,000 oocysts g(-1) of E. echidnae and remained clinically healthy during this study. Sub-adult and adult wild, and short-term captive, echidnas shed oocysts of both E. echidnae and E. tachyglossi . The lack of coccidia in juvenile short-beaked echidnas suggests these animals are probably non-immune and should not be placed in environments heavily contaminated with oocysts. In addition, no oocysts were found in captive long-beaked echidnas ( Zaglossus bartoni bartoni , n  =  2) housed at Taronga Zoo. This study represents an important step in understanding the host-parasite interaction between coccidia and short-beaked echidnas.

  8. Midlatitude land surface temperature impacts the timing and structure of glacial maxima

    NASA Astrophysics Data System (ADS)

    Thomas, Elizabeth K.; Clemens, Steven C.; Sun, Youbin; Huang, Yongsong; Prell, Warren; Chen, Guangshan; Liu, Zhengyu; Loomis, Shannon

    2017-01-01

    Orbitally modulated insolation is thought to be among the key mechanisms driving ice sheet change. Yet, late Pleistocene ice sheets attained maximum marginal extents prior to summer insolation minima, suggesting that additional mechanisms influenced ice growth and decay. Continuous orbital-scale terrestrial temperature records can help define the role that temperature near ice sheet margins plays on the timing and structure of glacial maxima. We hypothesize, based on a 360 kyr long air temperature record from the Chinese Loess Plateau and shorter records from North America and Europe, that midlatitude terrestrial temperature influenced the rate of ice sheet growth prior to and during glacial maxima; cold conditions prior to glacial maxima enhanced ice sheet growth, while warming during glacial maxima inhibited further growth, despite low summer insolation. Thus, the midlatitude surface energy budget may be an important component of understanding and modeling ice volume, particularly for intervals prior to and during glacial maxima, when ice sheet margins reached midlatitudes.

  9. Studies on coccidia species of genus Eimeria from domestic rabbit (Oryctolagus cuniculus domesticus L.) in Jeddah, Saudi Arabia.

    PubMed

    Toula, F H; Ramadan, H H

    1998-12-01

    Five Eimeria species were reported from domestic rabbits (Oryctolagus cuniculus domesticus L.) caught from Jeddah, Saudi Arabia. According to their prevalences, they were: Eimeria perfarans (65%), E. magna (45%), E. stiedae (25%), E. exigua (20%) and E. piriformis (10%). 90% of the examined rabbits were positive and mixed infection with two or three Eimeria species was most frequent. E. stiedae, E. piriformis and E. exigua were identified and recorded for the first time from rabbits in Saudi Arabia and are considered as new locality or geographical distribution.

  10. Long-term effect of toltrazuril on growth performances of dairy heifers and beef calves exposed to natural Eimeria zuernii and Eimeria bovis infections.

    PubMed

    Veronesi, Fabrizia; Diaferia, Manuela; Viola, Orfeo; Fioretti, Daniela Piergili

    2011-11-01

    The long-term effects of a toltrazuril treatment against natural Eimeria bovis and/or Eimeria zuernii infections were investigated in comparison with diclazuril and untreated controls on two dairy (Italian Friesian breed) and two beef (Chianina breed) farms. At each trial site, 30 calves were allocated into three groups of 10 calves each: T (treated with toltrazuril), D (treated with diclazuril) and C (left untreated). For 40 weeks post-treatment, the calves were weighed and examined clinically and parasitologically. The oocyst counts as well as the number of scour days were significantly lower in the T group than in the D and C groups. Final bodyweights and body condition scores of the T group exceeded those of groups C and D. The results confirmed that toltrazuril was highly efficacious, safe and provided productive benefits in dairy and beef calves.

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

    PubMed

    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

  12. Characterisation of full-length cDNA sequences provides insights into the Eimeria tenellatranscriptome

    PubMed Central

    2012-01-01

    Background Eimeria tenella is an apicomplexan parasite that causes coccidiosis in the domestic fowl. Infection with this parasite is diagnosed frequently in intensively reared poultry and its control is usually accorded a high priority, especially in chickens raised for meat. Prophylactic chemotherapy has been the primary method used for the control of coccidiosis. However, drug efficacy can be compromised by drug-resistant parasites and the lack of new drugs highlights demands for alternative control strategies including vaccination. In the long term, sustainable control of coccidiosis will most likely be achieved through integrated drug and vaccination programmes. Characterisation of the E. tenella transcriptome may provide a better understanding of the biology of the parasite and aid in the development of a more effective control for coccidiosis. Results More than 15,000 partial sequences were generated from the 5' and 3' ends of clones randomly selected from an E. tenella second generation merozoite full-length cDNA library. Clustering of these sequences produced 1,529 unique transcripts (UTs). Based on the transcript assembly and subsequently primer walking, 433 full-length cDNA sequences were successfully generated. These sequences varied in length, ranging from 441 bp to 3,083 bp, with an average size of 1,647 bp. Simple sequence repeat (SSR) analysis identified CAG as the most abundant trinucleotide motif, while codon usage analysis revealed that the ten most infrequently used codons in E. tenella are UAU, UGU, GUA, CAU, AUA, CGA, UUA, CUA, CGU and AGU. Subsequent analysis of the E. tenella complete coding sequences identified 25 putative secretory and 60 putative surface proteins, all of which are now rational candidates for development as recombinant vaccines or drug targets in the effort to control avian coccidiosis. Conclusions This paper describes the generation and characterisation of full-length cDNA sequences from E. tenella second generation

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

    PubMed

    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.

  14. Use of monoclonal antibodies developed against chicken coccidia (Eimeria) to study invasion and development of Eimeria reichenowi in Florida sandhill cranes (Grus canadensis)

    USGS Publications Warehouse

    Augustine, P.C.; Olsen, G.H.; Danforth, H.D.; Gee, G.F.; Novilla, M.

    2001-01-01

    Eimeria gruis and Eimeria reichenowi are common coccidial parasites of a number of species of cranes. Until recently, little was known about either the site for invasion or the dynamics of early development of the crane coccidia because of the difficulty of identifying sporozoites and early developmental stages of these parasites by conventional staining methods. In the present study, monoclonal antibodies (MAbs) elicited against Eimeria spp. of chickens and turkeys were found to cross-react with sporozoites and developmental stages of E. reichenowi in the tissues of Florida sandhill cranes (Grus canadensis). With these Mabs, E. reichenowi sporozoites were found in specimens taken at 6 hr postinoculation (PI) from just proximal to Meckel's diverticulum in the jejunum to the ileocecal juncture. Fewer were found in the ceca and rectum and none in the duodenal loop. At 24 hr PI, there were markedly fewer sporozoites and their location had shifted to the duodenum. No stages were seen in intestinal cells at 5 days PI (DPI), but trophozoites had developed in the liver and spleen. At 10 DPI, sexual stages were detected in the intestine from the duodenal loop through Meckel's diverticulum but not in other organs. By 14 DPI, numerous developmental stages were detected in the intestine (ceca and jejunum), liver, and lungs but not in the heart, kidney, or brain. The number, location, and maturity of the stages in the ceca differed markedly from those in the jejunum.

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

    PubMed

    Varghese, T

    1977-05-01

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

  16. Eimeria species parasites as novel vaccine delivery vectors: anti-Campylobacter jejuni protective immunity induced by Eimeria tenella-delivered CjaA.

    PubMed

    Clark, Julie D; Oakes, Richard D; Redhead, Keith; Crouch, Colin F; Francis, Michael J; Tomley, Fiona M; Blake, Damer P

    2012-03-30

    Vaccination of poultry against coccidiosis caused by the Eimeria species is almost entirely based upon varied formulations of live parasites. The recent development of a series of protocols that support genetic complementation by transfection in Eimeria now provides an opportunity to utilise live anticoccidial vaccines to deliver additional vaccinal antigens. The capacity of Eimeria tenella to express an exogenous antigen and induce an immune response during in vivo infection which is protective against subsequent bacterial challenge has been tested here using the anti-Campylobacter jejuni vaccine candidate CjaA. Using restriction enzyme mediated integration (REMI) a transgenic E. tenella population expressing CjaA and the fluorescent reporter mCitrine has been developed. Vaccination of specific pathogen free chickens by single or multiple oral inoculation of E. tenella-CjaA oocysts induced 91% and 86% immune protection against C. jejuni challenge compared with unvaccinated and wild-type E. tenella vaccinated controls (p<0.001). Increasing vaccination number had no significant influence on the magnitude of protection. These results support the hypothesis that eimerian parasites can be developed as multivalent vaccine vectors and encourage the extension of these studies.

  17. Comparison of Global Transcriptional Responses of Chicken Following Primary and Secondary Eimeria acervulina Infections

    USDA-ARS?s Scientific Manuscript database

    In the current study, we compared chicken gene transcriptional profiles following primary and secondary infections with Eimeria acervulina using a 9.6K avian intestinal intraepithelial lymphocyte cDNA microarray (AVIELA). Gene Ontology analysis showed that primary infection significantly modulated ...

  18. Prevalence of Eimeria (Apicomplexa: Eimeriidae) in reintroduced Gunnison's prairie dogs (Cynomys gunnisoni)

    Treesearch

    M. M. Ryan; K. H. Decker; D. W. Duszynski

    2001-01-01

    Fecal samples from 54 Gunnison's prairie dogs (Cynomys gunnisoni) from Albuquerque, NM were analyzed for the presence of coccidia and all were positive. They were then relocated to an abandoned prairiedog town on the Sevilleta Long Term Ecological Research (LTER) site. Six Eimeria species, E. callospermophili, E. cynomysis, E. pseudospermophili (new host record),...

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

    USDA-ARS?s Scientific Manuscript database

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

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

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

  2. Development of an in vitro assay for monensin and salinomycin sensitivity in Eimeria tenella

    USDA-ARS?s Scientific Manuscript database

    Applying ionophore drugs and synthetic chemicals to poultry feed has been used for over 50 years to control avian coccidiosis. While this strategy has been and continues to be effective, drug resistance has prompted alternatives such as drug rotation or vaccination with live Eimeria oocysts. Often...

  3. Seasonal recovery of Eimeria oocysts from soil on naturally contaminated pastures.

    PubMed

    Lassen, Brian; Lepik, Triin; Järvis, Toivo

    2014-03-01

    Though Eimeria is an important parasite of cattle, research is lacking on how the parasite persist in the pasture soils. In this study, feces samples were collected from three pastures in June and October 2010 and soil samples in April 2011. Coordinates of sampling locations were recorded with Global Positioning System together with information about grass cover, shade, and elevation. All soil samples were collected from the same locations as the fecal samples and used in model evaluating the possible factors influencing the concentration of oocysts in the soil. Feces and soil samples were investigated using a quantitative flotation technique. Eimeria oocysts were found in 95.6% of fecal samples collected in summer and 84.5% of samples in fall. In contrast, the same locations soil samples were positive for Eimeria oocysts in 37.3% (summer) and 44.3% (fall). Despite larger numbers of oocysts in fecal samples shed during summer compared to fall, there was no difference in the concentration of oocysts in soil samples the following spring. The odds of higher numbers of oocysts in soil samples in spring were higher if fecal samples collected in summer were in shade or if containing Eimeria alabamensis during the fall. Factors other than the concentrations of oocysts shed in feces appear to affect whether oocysts persist between grazing seasons.

  4. Efficacy of amprolium for the treatment of pathogenic Eimeria species in Boer goat kids.

    PubMed

    Young, Gabrielle; Alley, Mark L; Foster, Derek M; Smith, Geof W

    2011-06-10

    This study evaluated the efficacy of two different doses of amprolium in goats heavily infected with pathogenic Eimeria species. Forty Boer goat kids ranging from 3 to 5 months of age with naturally occurring coccidiosis were randomly divided into 2 groups and treated orally with amprolium at doses of 10mg/kg daily for 5 days (n=20) or 50mg/kg daily for 5 days (n=20). The Eimeria oocyst per gram concentrations were significantly reduced on day 7 in the kids that received amprolium at 50mg/kg, however oocyst concentrations were not significantly reduced in goats that received the 10mg/kg dose. Out of 100 Eimeria oocysts identified from a pooled fecal sample, E. christenseni was the most frequently identified (52%) coccidial species present. The results of this trial indicate that amprolium can be an effective treatment for pathogenic Eimeria species in goat kids, however higher and extralabel doses (50mg/kg) should be used. Copyright © 2011 Elsevier B.V. All rights reserved.

  5. Expression of innate immunity genes during Eimeria praecox infection in chickens

    USDA-ARS?s Scientific Manuscript database

    Intestinal colonization of avian species by Eimeria parasites results in the enteric disease, coccidiosis. A study was carried out to assess the immunological effects of E. praecox infection on the gut of infected chickens. In experiment 1, birds were orally gavaged with 50,000 E. praecox oocysts;...

  6. [Coccidiosis causes diarrhea in calves in the pasture. Pasture coccidiosis caused by Eimeria alabamensis].

    PubMed

    Snoep, J J; Potters, J B

    2004-03-01

    A severe outbreak of diarrhoea in young (age 18-20 month) cattle 14 days after their first turnout is described. In the manure large numbers Eimeria alabamensis oocysts were found. This infection is considered as a case of coccidial diarrhoea in (older) calves at pasture.

  7. Variation in Eimeria oocyst count and species composition in weanling beef heifers.

    PubMed

    Lucas, A S; Swecker, W S; Scaglia, G; Lindsay, D S; Zajac, A M

    2006-10-01

    Rectal fecal samples were collected daily on 10 consecutive days in November 2004 from 11 weaned beef heifers to assess daily variation in fecal oocyst count and species composition. Subsequent samples were collected from the same animals on 15 April 2005 and 9 June 2005. Oocyst numbers were determined by the modified McMaster's test, and species were identified by examination of oocysts recovered with the Wisconsin sugar flotation technique. Soil samples were collected from the heifer pasture on 8 June 2005, and oocysts were quantified and identified to species. Mean fecal oocyst counts varied little at all sampling dates ranging from 134-377 oocysts/g. Ten Eimeria spp. were identified in fecal samples collected in November and April and 11 in June. Eimeria bovis was the most common species identified at all samplings. Mean species composition showed little variation during the 10-day sampling period in November, remained similar in April, and varied slightly in June. Twelve Eimeria spp. were identified in soil samples in proportions similar to those seen in fecal samples. The results indicate that clinically normal weanling beef heifers are likely to be infected with a diverse, but relatively stable, community of Eimeria spp.

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

    PubMed

    Lassen, Brian; Viltrop, Arvo; Järvis, Toivo

    2009-10-01

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

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

    PubMed

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

    2011-10-01

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

  10. Enhanced egress of intracellular Eimeria tenella sporozoites by splenic lymphocytes from coccidia-infected chickens

    USDA-ARS?s Scientific Manuscript database

    Egress, which describes the mechanism that some intracellular parasites use to exit from parasitophorous vacuoles and host cells, plays a very important role in the parasite life cycle and is central to Eimeria propagation and pathogenesis. Despite the importance of egress in the intracellular paras...

  11. Characterization of monoclonal antibodies that recognize the Eimeria tenella microneme protein MIC2

    USDA-ARS?s Scientific Manuscript database

    The Apicomplexan pathogens of the species Eimeria cause coccidiosis, an intestinal disease of chickens, which has a major economic impact on the poultry industry. Members of the phylum Apicomplexa share an assortment of unique secretory organelles (rhoptries, micronemes and dense granules) that me...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    PubMed

    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.

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

    PubMed

    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

  15. Transgenic Eimeria magna Pérard, 1925 Displays Similar Parasitological Properties to the Wild-type Strain and Induces an Exogenous Protein-Specific Immune Response in Rabbits (Oryctolagus cuniculus L.).

    PubMed

    Tao, Geru; Shi, Tuanyuan; Tang, Xinming; Duszynski, Donald W; Wang, Yunzhou; Li, Chao; Suo, Jingxia; Tian, Xiuling; Liu, Xianyong; Suo, Xun

    2017-01-01

    Rabbit coccidiosis causes great economic losses to world rabbitries. Little work has been done considering genetic manipulation on the etiological agents, rabbit Eimeria spp. In this study, we constructed a transgenic line of Eimeria magna (EmagER) expressing enhanced yellow fluorescent protein (EYFP) and red fluorescent protein (RFP) using regulatory sequences of Eimeria tenella and Toxoplasma gondii. We observed the life cycle of EmagER and confirmed that the transgenic parasites express exogenous proteins targeted to different cellular compartments throughout the entire life cycle. EYFP was expressed mainly in the nucleus and RFP both in the nucleus and cytoplasm. Then, coccidia-free, laboratory-reared 40-day-old rabbits were primarily infected with either EmagER or wild-type strain oocysts and challenged with the wild-type strain. EmagER showed similar reproductivity and immunogenicity to the wild-type strain. Finally, we examined the foreign protein-specific immune response elicited by EmagER. Rabbits were immunized with either transgenic or wild-type oocysts. Immune response against parasite-soluble antigen, EYFP and RFP in spleen, and mesenteric lymph nodes were detected by quantitative real-time PCR. The relative expression level of IFN-γ, IL-2, and TNF-α were higher in EmagER-immunized rabbits than wild-type parasites-immunized rabbits after stimulation with EYFP and RFP. Our study confirmed that a specific immune response was induced by the exogenous protein expressed by EmagER and favored future studies on application of transgenic rabbit coccidia as recombinant vaccine vectors.

  16. Transgenic Eimeria magna Pérard, 1925 Displays Similar Parasitological Properties to the Wild-type Strain and Induces an Exogenous Protein-Specific Immune Response in Rabbits (Oryctolagus cuniculus L.)

    PubMed Central

    Tao, Geru; Shi, Tuanyuan; Tang, Xinming; Duszynski, Donald W.; Wang, Yunzhou; Li, Chao; Suo, Jingxia; Tian, Xiuling; Liu, Xianyong; Suo, Xun

    2017-01-01

    Rabbit coccidiosis causes great economic losses to world rabbitries. Little work has been done considering genetic manipulation on the etiological agents, rabbit Eimeria spp. In this study, we constructed a transgenic line of Eimeria magna (EmagER) expressing enhanced yellow fluorescent protein (EYFP) and red fluorescent protein (RFP) using regulatory sequences of Eimeria tenella and Toxoplasma gondii. We observed the life cycle of EmagER and confirmed that the transgenic parasites express exogenous proteins targeted to different cellular compartments throughout the entire life cycle. EYFP was expressed mainly in the nucleus and RFP both in the nucleus and cytoplasm. Then, coccidia-free, laboratory-reared 40-day-old rabbits were primarily infected with either EmagER or wild-type strain oocysts and challenged with the wild-type strain. EmagER showed similar reproductivity and immunogenicity to the wild-type strain. Finally, we examined the foreign protein-specific immune response elicited by EmagER. Rabbits were immunized with either transgenic or wild-type oocysts. Immune response against parasite-soluble antigen, EYFP and RFP in spleen, and mesenteric lymph nodes were detected by quantitative real-time PCR. The relative expression level of IFN-γ, IL-2, and TNF-α were higher in EmagER-immunized rabbits than wild-type parasites-immunized rabbits after stimulation with EYFP and RFP. Our study confirmed that a specific immune response was induced by the exogenous protein expressed by EmagER and favored future studies on application of transgenic rabbit coccidia as recombinant vaccine vectors. PMID:28167939

  17. Detailed study of anaerobic digestion of Spirulina maxima algae biomass

    SciTech Connect

    Samson, R.; LeDuy, A.

    1986-07-01

    Biomass of the blue-green alga Spirulina maxima was converted to methane using continuous stirred tank digesters with an energy conversion efficiency of 59%. Digesters were operated using once-a-day feeding with a retention time (theta) between 5 and 40 days, volatile solid concentrations (Sto) between 20 and 100 kg VS/cubic m, and temperatures between 15 and 52/sup 0/C. The results indicated a maximum methane yield of 0.35 cubic m (STP)/kg VS added at theta = 30 days and Sto = 20 kg VS/cubic m. Under such conditions, the energy conversion of the algal biomass to methane was 59%. The maximum methane production rate of 0.80 cubic m (STP)/cubic m day was obtained with theta = 20 days and Sto = 100 kg VS/cubic m. The mesophilic condition at 35/sup 0/C produced the maximum methane yield and production rate. The process was stable and characterized by a high production of volatile acids (up to 23,200 mg/l), alkalinity (up to 20,000 mg/l), and ammonia (up to 7000 mg/l), and the high protein content of the biomass produced a well-buffered environment which reduced inhibitory effects. At higher loading rates, the inhibition of methanogenic bacteria was observed, but there was no clear-cut evidence that such a phenomenon was due to nonionized volatile acids or gaseous ammonia. The kinetic analysis using the model proposed by Chen and Hashimoto indicated that the minimum retention time was seven days. The optimum retention time increased gradually from 11 to 16 days with an increase in the initial volatile solid concentration. The kinetic constant K decreased with the improvement in the digester performance and increased in parallel with the ammonia concentration in the culture media. 32 references.

  18. Inhibition of host cell apoptosis by Eimeria bovis sporozoites.

    PubMed

    Lang, Mirjam; Kann, Michael; Zahner, Horst; Taubert, Anja; Hermosilla, Carlos

    2009-03-09

    Sophisticated evasion strategies of obligate intracellular parasites, in particular prevention of host cell apoptosis, are necessary to ensure successful replication. To study the ability of Eimeria bovis in this regard, in vitro experiments were performed applying bovine foetal gastrointestinal cells (BFGC), bovine umbilical vein endothelial cells (BUVEC) and African green monkey kidney cells (VERO) as host cells. BUVEC and BFGC allow maturation of sporozoites to macromeronts, in VERO cells sporozoites survive for weeks without showing further development. In highly infected BUVEC monolayers, infected cells survived until merozoite release whereas uninfected cells underwent apoptosis. Light microscopy and TUNEL assays performed 3-10 days p.i. showed that, within infected BFGC and VERO cell monolayers, uninfected cells underwent programmed cell death after application of various inducers of apoptosis, whereas infected cells survived. Incidentally, the anti-apoptotic efficacies in infected cells were independent of the drugs and the host cell type. We could not demonstrate significant differences between infected and uninfected cells after colchicin treatment in terms of translation of phosphatidylserines to the host cell surface, caspase 3 activity and cytochrome c release, probably since obtainable infection rates were too low. However, we could show by laser scanning confocal microscopy on single cell levels that the expression of the anti-apoptotic factors cellular Flice inhibitory protein (c-FLIP) and cellular inhibition of apoptosis protein 1 (c-IAP1) were enhanced in E. bovis infected cells after application of colchicin, in the latter case also in non-infected cells directly neighbouring infected ones. Our data show that E. bovis protects its host cell from apoptosis by increasing expression of c-IAP1 and c-FLIP.

  19. Nitric oxide production during Eimeria tenella infections in chickens.

    PubMed

    Allen, P C

    1997-06-01

    The objective of this study was to gather evidence for production of nitric oxide (NO) during a primary infection with the protozoan parasite Eimeria tenella, which carries out its life cycle in the ceca of chickens. Relationships of plasma levels of NO2(-)+NO3-, stable metabolites of NO, with parasite dose and with time after infection were examined, as well as effects of administration of aminoguanidine, an inhibitor of induced nitric oxide synthase (iNOS). Inoculation with 5 x 10(4) and 1 x 10(6) but not 1 x 10(3) oocysts per chick caused significant (P < or = 0.05) increases in micromolar concentrations of plasma NO3(-)+NO3- when measured at 7 d postinoculation (PI). In chickens inoculated with 5 x 10(4) oocysts, significant (P < or = 0.05) increases in plasma NO2(-)+NO3- were seen at 5 and 7 but not 3 d PI. Daily intraperitoneal administration of 1.25 mg per chick aminoguanidine during the period of infection did not lower the increases in plasma NO2(-)+NO3- seen at 5 and 7 d PI, and did not affect the degree of colonization of the cecal tissue by the parasite. However, administration of aminoguanidine did alter the gross appearance of the ceca, which were less swollen and filled with blood at 5 and 7 d PI as compared with ceca from untreated chickens. Hemorrhage is a major pathological manifestation of E. tenella infections, associated with the disruption of the cecal mucosa by the developing parasite. The results of this experiment are consistent with the hypothesis that an aminoguanidine-inhibitable NO synthase, perhaps in the vascular endothelium of the cecal blood vessels, may contribute to hemorrhage by causing vasodilation.

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

    PubMed

    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.

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

    PubMed

    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.

  2. Virus-like particles in Eimeria nieschulzi are associated with multiple RNA segments.

    PubMed

    Roditi, I; Wyler, T; Smith, N; Braun, R

    1994-02-01

    RNA preparations from sporulated oocysts of Eimeria nieschulzi were found to contain 2 double-stranded RNA segments of 5.0 kb and 5.7 kb that were not present in other species of Eimeria. Treatment of crude lysates with RNase A revealed that in addition to these two segments, 3 other segments of 0.57 kb, 0.72 kb and 11.5 kb were protected from digestion, suggesting that they were enclosed within particles. Virus-like particles with a diameter of approximately 39 nm were purified by caesium chloride buoyant density centrifugation. Four of the five RNA segments copurified with these particles. In keeping with the nomenclature generally adopted for protozoan viruses, we have named this new isolate ENV 1. The largest RNA segment does not cosediment with ENV 1 particles and may be derived from another RNA-protein complex that is unstable under the conditions used. The particle size and genome structure of ENV 1 both differ from that of the Eimeria stiedae virus (ESV), which is the only other virus to have been isolated from Eimeria to date. Short cDNA clones derived from ENV 1 show significant homology to a region of the Leishmania virus (LRV 1) genome that encodes an RNA-dependent RNA polymerase. The polymerase sequences from ENV 1 and LRV 1 are more closely related to each other than to any other protein sequences in the GenEMBL Database. This raises intriguing questions about the origins of the two viruses, since Eimeria and Leishmania normally infect different hosts and also show different cell tropisms within these hosts.

  3. Enhanced Egress of Intracellular Eimeria tenella Sporozoites by Splenic Lymphocytes from Coccidian-Infected Chickens▿

    PubMed Central

    Dong, Xiaojuan; Abdelnabi, Ghada H.; Lee, Sung H.; Li, Guangxing; Jin, Hong; Lillehoj, Hyun S.; Suo, Xun

    2011-01-01

    Egress, which describes the mechanism that some intracellular parasites use to exit from parasitophorous vacuoles and host cells, plays a very important role in the parasite life cycle and is central to Eimeria propagation and pathogenesis. Despite the importance of egress in the intracellular parasite's life cycle, very little information is known on this process compared to other steps, e.g., invasion. The present study was conducted to investigate the interplay between the host adaptive immune system and Eimeria egression. Splenic lymphocytes or soluble immune factors were incubated with parasite-infected host cells for 3 or 5 h, and the percentage of egress was calculated according to an established formula. Viability of egressed parasites and host cells was tested using trypan blue exclusion and annexin V and propidium iodide staining, respectively. We found that premature egression of sporozoites from Eimeria tenella-infected primary chicken kidney cells or from chicken peripheral blood mononuclear cells occurred when the cells were cocultured in vitro with spleen lymphocytes from E. tenella-infected chickens but not when they were cocultured with splenocytes from uninfected chickens. Eimeria-specific antibodies and cytokines (gamma interferon [IFN-γ], interleukin-2 [IL-2], and IL-15), derived from E. tenella-primed B and T lymphocytes, respectively, were capable of promoting premature egress of sporozoites from infected host cells. Both egressed parasites and host cells were viable, although the latter showed reduced reinvasion ability. These results suggest a novel, immune-mediated mechanism that the host exploits to interrupt the normal Eimeria life cycle in vivo and thereby block the release of mature parasites into the environment. PMID:21628515

  4. The effect of the mitochondrial permeability transition pore on apoptosis in Eimeria tenella host cells.

    PubMed

    Xu, Zhi-Yong; Zheng, Ming-Xue; Zhang, Yan; Cui, Xiao-Zhen; Yang, Sha-Sha; Liu, Rui-Li; Li, Shan; Lv, Qiang-Hua; Zhao, Wen-Long; Bai, Rui

    2016-10-01

    Although the mitochondrial permeability transition pore (MPTP) is associated with cellular apoptosis and necrosis, its effect in host response to Eimeria infections is not well understood. In an effort to better understand the effect of MPTP on apoptosis in Eimeria tenella host cells, an MPTP inhibitor (cyclosporin A) was used to inhibit MPTP opening in vitro. Cecal epithelial cells from chick embryos, which were either treated or non-treated with cyclosporin A, were used as Eimeria tenella host cells. In addition, primary chick embryo cecum epithelial cell culture techniques and flow cytometry were used to detect the dynamic changes in MPTP opening, mitochondrial transmembrane potential, and cell apoptosis rate of Eimeria tenella host cells. Compared with the control group, cytometric techniques showed that untreated host cells exhibited a significantly higher (P < 0.01) degree of MPTP opening but lower (P < 0.01 or P < 0.05) mitochondrial transmembrane potential. Moreover, untreated group cells had less apoptosis (P < 0.01) at 4 h and more apoptosis (P < 0.05 or P < 0.01) at 24 to 120 h as compared with control group cells. After the application of cyclosporin A, the degree of MPTP opening in the treated group was significantly lower (P < 0.01) at 4 to 120 h compared to the untreated group, whereas the treated group had higher (P < 0.05 or P < 0.01) mitochondrial transmembrane potentials at 24 to 120 h. Flow cytometry assays also showed that there was less (P < 0.05 or P < 0.01) apoptosis after 24 h in the treated group than in the untreated group. Taken together, these observations indicate that MPTP is a key node that plays a predominant role in the mitochondrial apoptosis pathway in the host cell induced by Eimeria tenella.

  5. Enhanced egress of intracellular Eimeria tenella sporozoites by splenic lymphocytes from coccidian-infected chickens.

    PubMed

    Dong, Xiaojuan; Abdelnabi, Ghada H; Lee, Sung H; Li, Guangxing; Jin, Hong; Lillehoj, Hyun S; Suo, Xun

    2011-08-01

    Egress, which describes the mechanism that some intracellular parasites use to exit from parasitophorous vacuoles and host cells, plays a very important role in the parasite life cycle and is central to Eimeria propagation and pathogenesis. Despite the importance of egress in the intracellular parasite's life cycle, very little information is known on this process compared to other steps, e.g., invasion. The present study was conducted to investigate the interplay between the host adaptive immune system and Eimeria egression. Splenic lymphocytes or soluble immune factors were incubated with parasite-infected host cells for 3 or 5 h, and the percentage of egress was calculated according to an established formula. Viability of egressed parasites and host cells was tested using trypan blue exclusion and annexin V and propidium iodide staining, respectively. We found that premature egression of sporozoites from Eimeria tenella-infected primary chicken kidney cells or from chicken peripheral blood mononuclear cells occurred when the cells were cocultured in vitro with spleen lymphocytes from E. tenella-infected chickens but not when they were cocultured with splenocytes from uninfected chickens. Eimeria-specific antibodies and cytokines (gamma interferon [IFN-γ], interleukin-2 [IL-2], and IL-15), derived from E. tenella-primed B and T lymphocytes, respectively, were capable of promoting premature egress of sporozoites from infected host cells. Both egressed parasites and host cells were viable, although the latter showed reduced reinvasion ability. These results suggest a novel, immune-mediated mechanism that the host exploits to interrupt the normal Eimeria life cycle in vivo and thereby block the release of mature parasites into the environment.

  6. Recombinant allergens

    PubMed Central

    Jutel, Marek; Solarewicz-Madejek, Katarzyna; Smolinska, Sylwia

    2012-01-01

    Allergen specific immunotherapy (SIT) is the only known causative treatment of allergic diseases. Recombinant allergen-based vaccination strategies arose from a strong need to both to improve safety and enhance efficacy of SIT. In addition, new vaccines can be effective in allergies including food allergy or atopic dermatitis, which poorly respond to the current treatment with allergen extracts. A number of successful clinical studies with both wild-type and hypoallergenic derivatives of recombinant allergens vaccines have been reported for the last decade. They showed high efficacy and safety profile as well as very strong modulation of T and B cell responses to specific allergens. PMID:23095874

  7. Alleviation of metabolic abnormalities induced by excessive fructose administration in Wistar rats by Spirulina maxima.

    PubMed

    Jarouliya, Urmila; Zacharia, J Anish; Kumar, Pravin; Bisen, P S; Prasad, G B K S

    2012-03-01

    Diabetes mellitus is a metabolic disorder characterized by hyperglycaemia. Several natural products have been isolated and identified to restore the complications of diabetes. Spirulina maxima is naturally occurring fresh water cyanobacterium, enriched with proteins and essential nutrients. The aim of the study was to determine whether S. maxima could serve as a therapeutic agent to correct metabolic abnormalities induced by excessive fructose administration in Wistar rats. Oral administration of 10 per cent fructose solution to Wistar rats (n = 5 in each group) for 30 days resulted in hyperglycaemia and hyperlipidaemia. Aqueous suspension of S. maxima (5 or 10%) was also administered orally once daily for 30 days. The therapeutic potential of the preparation with reference to metformin (500 mg/kg) was assessed by monitoring various biochemical parameters at 10 day intervals during the course of therapy and at the end of 30 days S. maxima administration. Significant (P<0.001) reductions in blood glucose, lipid profile (triglycerides, cholesterol and LDL, VLDL) and liver function markers (SGPT and SGOT) were recorded along with elevated level of HDL-C at the end of 30 days therapy of 5 or 10 per cent S. maxima aquous extract. Co-administration of S. maxima extract (5 or 10% aqueous) with 10 per cent fructose solution offered a significant protection against fructose induced metabolic abnormalities in Wistar rats. The present findings showed that S. maxima exhibited anti-hyperglycaemic, anti-hyperlipidaemic and hepatoprotective activity in rats fed with fructose. Further studies are needed to understand the mechanisms.

  8. [Comparison of the proteome of the sporutated oocysts of Eimeria tenella diclazuril sensitive strain with diclazuril resistant strain].

    PubMed

    Jiang, Lian-Lian; Huang, Bing; Han, Hong-Yu; Zhao, Qi-Ping; Dong, Hui; Chen, Zhao-Guo

    2005-05-01

    Two-dimensional electrophoresis (2-DE) was employed to compare the proteome of Diclazuril-resistance Eimeria tenella with that of sensitive strains for identifying unique proteins of these stains. 5 protein spots were found to express differentially. Four spots which remarkably were measured by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The data obtained from peptide mass fingerprinting were used in NCBInr database search, two protein spots in gel were identified as Eimeria tenella sporulated oocyst TA4 antigen protein, Heat shock 70kD protein, two protein spots were functional proteins of Eukaryote. These proteins are potentially basic work for finding molecular mechanism about drug-resistance of Eimeria tenella and new marker in the detection of resistance of Eimeria tenella.

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

    USDA-ARS?s Scientific Manuscript database

    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...

  10. Efficacy of toltrazuril 5 % suspension against Eimeria bovis and Eimeria zuernii in calves and observations on the associated immunopathology.

    PubMed

    Jonsson, Nicholas N; Piper, Emily K; Gray, Christian P; Deniz, Abdulkerim; Constantinoiu, Constantin C

    2011-08-01

    16 Calves were each infected with suspensions containing a mixture of approximately 230,000 Eimeria bovis and 70,000 E. zuernii oocysts, which resulted in detection of oocysts in faeces of 12 of 16 calves by day +14 after infection. On day +14 after infection calves were either treated (n = 8) with toltrazuril at 15 mg/kg body weight or with a placebo. Observations were made on the clinical condition, faecal score and liveweight of calves daily from one day post infection (pi) until 24 days pi when all calves were euthanised and examined post mortem. Samples were collected from ileum and colon for histological, immunohistochemical and gene expression studies. The study demonstrated an efficacy of toltrazuril for the treatment of E. bovis and E. zuernii infections in calves reaching 99 % (based on arithmetic mean oocyst counts in faeces) within three days of treatment and remaining at or above this level for six days. Toltrazuril did not have a significant effect on the pattern and extent of immune cellular infiltration in the mucosa of ileum and colon, but the expression of the genes coding IL-2, IL-10 and TNF-α in the ileum and TNF-α in the colon were elevated in calves treated with toltrazuril. Higher levels of oocyst shedding were significantly associated with lower expression of genes coding for IL-2, IL-10 and higher IP-10. It is concluded that toltrazuril is effective for the treatment of coccidiosis due to E. bovis and E. zuernii in calves and enables the development of a normal or enhanced immune response to infection.

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

    PubMed

    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.

  12. Spectrum Recombination.

    ERIC Educational Resources Information Center

    Greenslade, Thomas B., Jr.

    1984-01-01

    Describes several methods of executing lecture demonstrations involving the recombination of the spectrum. Groups the techniques into two general classes: bringing selected portions of the spectrum together using lenses or mirrors and blurring the colors by rapid movement or foreshortening. (JM)

  13. Recombinant gonadotropins.

    PubMed

    Lathi, R B; Milki, A A

    2001-10-01

    Recombinant DNA technology makes it possible to produce large amounts of human gene products for pharmacologic applications, supplanting the need for human tissues. The genes for the alpha and beta subunits of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and human chorionic gonadotropin (hCG) have been characterized and cloned. Recombinant FSH (rFSH) has been shown to be safe and effective in the treatment of fertility disorders. In comparison with the urinary gonadotropin products, human menopausal gonadotropins (HMG), and urinary follitropins (uFSH), rFSH is more potent and better tolerated by patients. Recombinant HCG appears to be as efficacious as urinary HCG with the benefit of improved local tolerance. Recombinant LH (rLH) is likely to be recommended as a supplement to rFSH for ovulation induction in hypogonadotropic women. It may also benefit in vitro fertilization patients undergoing controlled ovarian hyperstimulation with rFSH combined with pituitary suppression, with a gonadotropin-releasing hormone agonist or antagonist.

  14. Spectrum Recombination.

    ERIC Educational Resources Information Center

    Greenslade, Thomas B., Jr.

    1984-01-01

    Describes several methods of executing lecture demonstrations involving the recombination of the spectrum. Groups the techniques into two general classes: bringing selected portions of the spectrum together using lenses or mirrors and blurring the colors by rapid movement or foreshortening. (JM)

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

    PubMed

    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.

  16. Pumpkin Seed Oil Extracted From Cucurbita maxima Improves Urinary Disorder in Human Overactive Bladder

    PubMed Central

    Nishimura, Mie; Ohkawara, Tatsuya; Sato, Hiroji; Takeda, Hiroshi; Nishihira, Jun

    2014-01-01

    The pumpkin seed oil obtained from Cucurbita pepo has been shown to be useful for the treatment of nocturia in patients with urinal disorders in several western countries. In this study, we evaluated the effect of the pumpkin seed oil from Cucurbita maxima on urinary dysfunction in human overactive bladder (OAB). Forty-five subjects were enrolled in this study. An extract of pumpkin seed oil from C. maxima (10 g of oil/day) was orally administrated for 12 weeks. After 6 and 12 weeks, urinary function was evaluated using Overactive Bladder Symptom Score (OABSS). Pumpkin seed oil from C. maxima significantly reduced the degree of OABSS in the subjects. The results from our study suggest that pumpkin seed oil extracts from C. maxima as well as from C. pepo are effective for urinary disorders such as OAB in humans. PMID:24872936

  17. Pumpkin Seed Oil Extracted From Cucurbita maxima Improves Urinary Disorder in Human Overactive Bladder.

    PubMed

    Nishimura, Mie; Ohkawara, Tatsuya; Sato, Hiroji; Takeda, Hiroshi; Nishihira, Jun

    2014-01-01

    The pumpkin seed oil obtained from Cucurbita pepo has been shown to be useful for the treatment of nocturia in patients with urinal disorders in several western countries. In this study, we evaluated the effect of the pumpkin seed oil from Cucurbita maxima on urinary dysfunction in human overactive bladder (OAB). Forty-five subjects were enrolled in this study. An extract of pumpkin seed oil from C. maxima (10 g of oil/day) was orally administrated for 12 weeks. After 6 and 12 weeks, urinary function was evaluated using Overactive Bladder Symptom Score (OABSS). Pumpkin seed oil from C. maxima significantly reduced the degree of OABSS in the subjects. The results from our study suggest that pumpkin seed oil extracts from C. maxima as well as from C. pepo are effective for urinary disorders such as OAB in humans.

  18. Cloning and characterization of three Eimeria tenella lipid phosphate phosphatases.

    PubMed

    Guo, Aijiang; Cai, Jianping; Luo, Xuenong; Zhang, Shaohua; Hou, Junling; Li, Hui; Cai, Xuepeng

    2015-01-01

    Although lipid phosphate phosphatases (LPPs) play an important role in cellular signaling in addition to lipid biosynthesis, little is thus far known about parasite LPPs. In this study, we characterized three Eimeria tenella cDNA clones encoding LPP named EtLPP1, EtLPP2 and EtLPP3. Key structural features previously described in LPPs, including the three conserved domains proposed as catalytic sites, a single conserved N-glycosylation site, and putative transmembrane domains were discovered in the three resulting EtLPP amino acid sequences. Expression of His6-tagged EtLPP1, -2, and -3 in HEK293 cells produced immunoreactive proteins with variable molecular sizes, suggesting the presence of multiple forms of each of the three EtLPPs. The two faster-migrating protein bands below each of the three EtLPP proteins were found to be very similar to the porcine 35-kDa LPP enzyme in their molecular size and the extent of their N-glycosylation, suggesting that the three EtLPPs are partially N-glycosylated. Kinetic analyses of the activity of the three enzymes against PA, LPA, C1P and S1P showed that Km values for each of the substrates were (in μM) 284, 46, 28, and 22 for EtLPP1; 369, 179, 237, and 52 for EtLPP2; and 355, 83, and 260 for EtLPP3. However, EtLPP3 showed negligible activity on S1P. These results confirmed that the three EtLPPs have broad substrate specificity. The results also indicated that despite structural similarities, the three EtLPPs may play distinct functions through their different models of substrate preference. Furthermore, particularly high expression levels of the three EtLPP genes were detected in the sporozoite stage of the E. tenella life cycle (p<0.001), suggesting that their encoded proteins might play an important biological function in the sporozoite stage.

  19. Cloning and sequencing of the allophycocyanin genes from Spirulina maxima (Cyanophyta)

    NASA Astrophysics Data System (ADS)

    Qin, Song; Hiroyuki, Kojima; Yoshikazu, Kawata; Shin-Ichi, Yano; Zeng, Cheng-Kui

    1998-03-01

    The genes coding for the α-and β-subunit of allophycocyanin ( apcA and apcB) from the cyanophyte Spirulina maxima were cloned and sequenced. The results revealed 44.4% of nucleotide sequence similarity and 30.4% of similarity of deduced amino acid sequence between them. The amino acid sequence identities between S. maxima and S. platensis are 99.4% for α subunit and 100% for β subunit.

  20. Implementation of Spectral Maxima Sound Processing for Cochlear Implants by Using Bark Scale Frequency Band Partition

    DTIC Science & Technology

    2007-11-02

    1 Implementation of Spectral Maxima Sound processing for cochlear implants by using Bark scale Frequency band partition Han xianhua1 Nie...new method on the basis of Bark scale frequency-band partition was presented to improve the recognition performance of cochlear implants . In the...nature of physics, it consists with human’s cochlea filter properties. Also the mechanism of a cochlear implant and its spectral maxima sound

  1. Construction of Eimeria tenella multi-epitope DNA vaccines and their protective efficacies against experimental infection.

    PubMed

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

    2015-08-15

    The search for effective vaccines against chicken coccidiosis remains a challenge because of the complex organisms with multiple life cycle stages of Eimeria. Combination of T-cell epitopes from different stages of Eimeria life cycle could be an optimal strategy to overcome the antigen complexity of the parasite. In this study, 4 fragments with concentrated T-cell epitopes from the sporozoite antigen SO7 and the merozoite antigen MZ5-7 of Eimeria tenella were cloned into eukaryotic expression vector pVAX1 in different forms, with or without chicken cytokines IL-2 or IFN-γ genes as genetic adjuvants, to construct multistage, multi-epitope DNA vaccines against Eimeria tenella. Transcription and expression of the multi-epitope DNA vaccines in vivo were detected by reverse transcription-PCR (RT-PCR) and Western blot. On the basis of survival rate, lesion score, body weight gain, oocyst decrease ratio and the anti-coccidial index (ACI), Animal experiments were carried out to evaluate the protective efficacy against Eimeria tenella. Results showed the constructed DNA vaccines were transcribed and translated successfully in vivo. Animal experiment showed that the multi-epitopes DNA vaccines were more effective to stimulate immune response than single fragment. Compared with the DNA vaccines composed with less T-cell epitopes, DNA vaccine pVAX1-m1-m2-s1-s2 containing 4 fragments with concentrated T-epitopes provided the highest ACI of 180.39. DNA vaccines composed of antigens from two developmental stages were more effective than the single-stage ones. Especially DNA vaccine pVAX1-m1-m2-s1-s2 provided the most effective protection with the ACI of 180.39. Furthermore, cytokines IL-2 or IFN-γ could improve the efficacy of the multi-epitope DNA vaccines significantly. Overall, pVAX1-m1-m2-s1-s2-IFN-γ provided the most effective protection with the ACI of 189.92. The multi-epitope DNA vaccines revealed in this study provide new candidates for Eimeria vaccine development.

  2. [Incorporation of a spiruline (Spirulina maxima) in swine food].

    PubMed

    Fevrier, C; Seve, B

    1975-01-01

    Two experiments on early weaned piglets and one experiment on sows during the first two reproductive cycles were made to determine the nutritive value of Spirulina maxima algae in the pig, taken as a "nutritional model" for man. The algae were introduced into the diets of the piglets, between 12 and 42 days of age, in a proportion representing 12 p. 100 of the total proteins. This level corresponded to 12 p. 100 algae in the first age feed (up to 21 days) in replacement of skim milk, and 8 p. 100 of the second age feed in replacement of soyabean meal. For the overall experimental period, from 12 to 42 days, the apparent digestibilities of the diets, measured by the chronium oxide marking, decreased by 6.9 and 6.1 points, respectively, for energy and total crude proteins as compared to the control diet. Replacement of the same raw materials by a soluble fish protein concentrate only led to a reduction of 2.9 and 3.5 points for energy and proteins. However, during the first week following weaning, the fish concentrate and spirulina gave identical results which were lower than those of the control (fig. 1). In spite of this lowering in the apparent digestibility the growths of the piglets were satisfactory and equivalent for all groups. Thus, the mĕtabolic utilisation of the fraction of feed absorbed was better for the spirulina group than for the control group, notably during the period between 12 to 21 days, although the supply of lysine in the spirulina group was 12 p. 100 lower (tab. VII). Consequently, this partially compensates for the low apparent digestibility of the algae in the young animal. For the female, the spirulina algaes were continously administered in a proportion of 5 p. 100 of the diet from a live weight of 75 kg. This did not change either the age or the weight at onset of puberty (tab IX). During the first reproductive cycle, the net weight gain of the sows receiving spirulina was lower than that of the control sows, but the number of piglets

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

    PubMed

    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.

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

    PubMed

    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

  5. Occurrences of Eimeria spp. and gastrointestinal nematodes in dairy calves in southern Minas Gerais, Brazil.

    PubMed

    Bruhn, Fábio Raphael Pascoti; Silva Júnior, Fidelis Antônio; Carvalho, André Henrique de Oliveira; Orlando, Débora Ribeiro; Rocha, Christiane Maria Barcellos Magalhães da; Guimarães, Antônio Marcos

    2012-01-01

    The aim of this cross-sectional observational study was to determine the frequency and factors associated with infection by Eimeria spp. and gastrointestinal nematodes in 356 calves on 20 dairy farms located in southern Minas Gerais, Brazil. Ten species of Eimeria spp. were identified, of which E. bovis (37.6%) and E. zuernii (17.9%) were the most frequent. From fecal cultures, four genera of gastrointestinal nematodes were recovered, of which Cooperia spp. (74.6%) and Haemonchus (19.4%) were the most frequent. Variables relating to higher levels of technology used on dairy farms showed a significant association (p < 0.05) with higher OPG and EPG counts, and are discussed in this study.

  6. In vitro inhibition of Eimeria tenella invasion by indigenous chicken Lactobacillus species.

    PubMed

    Tierney, J; Gowing, H; Van Sinderen, D; Flynn, S; Stanley, L; McHardy, N; Hallahan, S; Mulcahy, G

    2004-07-14

    The aim of this study was to determine the effects of indigenous chicken Lactobacillus species isolates from different parts of the gastrointestinal tract on Eimeria tenella invasion in vitro and to characterise the nature of inhibition, if any. The effects of competitive exclusion, steric interference and bacterial extracellular factors on E. tenella invasion were examined in an MDBK cell model. Several Lactobacillus species were initially isolated from chickens and identified by biochemical characteristics and 16S-rRNA. All Lactobacillus species isolates tested, significantly inhibited E. tenella invasion. Steric interference did not affect parasite invasion. Extracellular metabolic factors secreted by Lactobacillus species isolates into the surrounding media were shown to inhibit parasite invasion and these factors appeared to be heat stable. These results show that the natural microflora of poultry can provide a source of E. tenella-inhibiting Lactobacillus species in vitro, and thus may contribute to the control of Eimeria infection.

  7. Alternative mechanism of Eimeria bovis sporozoites to invade cells in vitro by breaching the plasma membrane.

    PubMed

    Behrendt, J H; Clauss, W; Zahner, H; Hermosilla, C

    2004-10-01

    In vitro Eimeria bovis sporozoites invade a wide range of cell types, and in the case of bovine cells, they may develop to first-generation schizonts. Often, however, they subsequently leave their host cell to invade a new one, which seems contrary to the classical way of infecting a cell by forming a parasitophorous vacuole. Using a standard, "cell wound assay," we show that E. bovis can invade bovine endothelial cells by breaching the plasma membrane and may again leave the surviving cell. Eimeria bovis sporozoites also infected VERO and HT29 cells but obviously without damaging the plasma membrane. The same held true when bovine endothelial cells were exposed to tachyzoites of Toxoplasma gondii and Neospora caninum. According to a literature report dealing with Plasmodium yoelii sporozoites, breaching the membrane of certain host cells may be a common phenomenon for coccidian sporozoites but may not be for merozoites.

  8. Calcium-dependent microneme protein discharge and in vitro egress of Eimeria tenella sporozoites.

    PubMed

    Yan, Xinlei; Tao, Geru; Liu, Xianyong; Ji, Yongsheng; Suo, Xun

    2016-11-01

    Egress is a vital step in the endogenous development of apicomplexan parasites, as it assures the parasites exit from consumed host cells and entry into fresh ones. However, little information has previously been reported on this step of Eimeria spp. In this study, we investigated in vitro egress of Eimeria tenella sporozoites triggered by acetaldehyde. We found that addition of exogenous acetaldehyde induces egress of sporozoites from primary chicken kidney cells (PCKs) and stimulate secretion of E. tenella microneme 2 protein (EtMic 2). Moreover, by using cellular calcium inhibitors, we further proved that these processes were dependent on the intracellular calcium of the parasites. Our findings provide clues to the study of interaction between eimerian parasites and their hosts.

  9. Kinetics of interleukin-2 production in chickens infected with Eimeria tenella.

    PubMed

    Miyamoto, Tadashi; Min, Wongi; Lillehoj, Hyun S

    2002-05-01

    Interleukin (IL)-2 is a major cytokine of cell-mediated immunity (CMI). Because chickens infected with Eimeria, the causative agent of coccidiosis, develop a robust cell-mediated response against the parasite, we measured IL-2 concentrations in vivo and in vitro during the course of primary and secondary experimental Eimeria tenella infections. IL-2 levels in serum and culture supernatants of spleen lymphocytes stimulated with mitogen or E. tenella sporozoites were significantly increased on day 7 post-primary infection compared with control group. This peak in IL-2 coincided with the time of maximum intestinal lesions as measured by cecum lesion scores. By contrast, during secondary infection highest IL-2 concentrations preceded intestinal lesions by 5 days (day 2 versus day 7, respectively). These results confirmed that IL-2 production is augmented during experimental coccidiosis and suggested that cellular immunity elicited during an anamnestic response to parasite reinfection is mediated, at least in part, by IL-2.

  10. Interleukin-2 production in SC and TK chickens infected with Eimeria tenella.

    PubMed

    Li, Guangxing; Lillehoj, Erik P; Lillehoj, Hyun S

    2002-01-01

    SC and TK inbred chicken strains display differential protective immunity to coccidiosis, SC being more resistant and TK susceptible to disease. In this study, the association between interleukin (IL)-2 and disease phenotype was assessed by cytokine quantification in serum, duodenum, cecum, and spleen cell cultures of SC and TK chickens experimentally infected with Eimeria tenella. In general, after primary infection, SC and TK strains produced equivalent amounts of IL-2 in all sources examined. However, after secondary infection, SC animals displayed significantly greater IL-2 levels in serum and the duodenum compared with strain TK. IL-2 production after reinfection with Eimeria may be an important factor contributing to the genetic differences in coccidiosis between SC and TK chickens and provides a rational foundation for cytokine-based immunotherapeutic approaches to disease control strategies.

  11. Ultrastructural studies of the effects of amprolium and dinitolmide on Eimeria acervulina macrogametes.

    PubMed

    Ball, S J; Pittilo, R M; Norton, C C; Joyner, L P

    1987-01-01

    Medication of chicks with 125 ppm amprolium or dinitolmide adversely affected oocyst sporulation of Eimeria acervulina (Weybridge strain). Dinitolmide delayed oocyst production and no oocyst wall formation was seen up to 168 h post infection. Both drugs caused large numbers of abnormally small wall-forming bodies to be produced in the macrogametes. In amprolium-fed chicks, abnormal oocyst wall formation was seen. It was concluded that the main drug action was against wall forming bodies of type 2.

  12. Estimation of the economical effects of Eimeria infections in Estonian dairy herds using a stochastic model.

    PubMed

    Lassen, Brian; Ostergaard, Søren

    2012-10-01

    In this study, a stochastic predictive model stimulating a constant infection pressure of Eimeria was used to estimate production outcome, economic, and effects of treatment decisions in a dairy herd of 100 cows. The intestinal parasite cause problems mainly in calves, and is known to have long term effects on the growth rate, and in severe cases can result in mortalities. Due to the inconspicuous nature of the parasite, the clinical signs and sub-clinical manifestations it may produce can be overlooked. Acquired data from literature and Estonian dairy farms were implemented in the SimHerd IV model to simulate three scenarios of symptomatic treatment: no calves treated (NT), default estimate of the current treatment strategy (DT), and all calves treated (AT). Effects of metaphylactic treatment were studied as a lowering of the infection pressure. Delay in the age for beginning of insemination of heifers was the effect with the largest economic impact on the gross margin, followed by calf mortality and reduction in growth rate. Large expenses were associated with the introduction of replacement heifers and feeding of heifers as a result of the delay in reaching a specific body weight at calving. Compared to the control scenarios, with no effects and treatments of Eimeria, dairy farmers were estimated to incur annual losses ranging 8-9% in the balanced income. Providing metaphylactic drugs resulted in an increased gross margin of 6-7%. Purchase of new heifers compensated for some production losses that would otherwise have enhanced expenses related to Eimeria. The simulation illustrates how effects of Eimeria infections can have long lasting impact on interacting management factors. It was concluded that all three simulated symptomatic treatment regimes provided only small economic benefits if they were applied alone and not in combination with lowering of infection pressure.

  13. Large-scale survey of the prevalence of Eimeria infections in domestic rabbits in China.

    PubMed

    Jing, Fa; Yin, Guangwen; Liu, Xianyong; Suo, Xun; Qin, Yinghe

    2012-04-01

    The current study examined the prevalence of Eimeria infections in domestic rabbits in China. A total of 480 faecal samples were collected from 48 farms in 14 provinces of China. Each faecal sample was subjected to oocyst counting and oocyst isolation. The Eimeria species from samples containing isolated and sporulated oocysts were morphologically identified under microscope. The overall prevalence of infections was 41.9% (201/480). Northwest China had the highest prevalence (70%), followed closely by Northeast China (65%) and Southwest China (62.5%). The prevalences in North China (34%) and South China (25.8%) were significantly lower. The large and medium farms had lower prevalences (34.2% and 37.2%, respectively) than the small farms (61.4%). Coccidian oocysts were found in 42.2% (76/180) of faecal samples from meat rabbits, 40% (28/70) from angora rabbits and 44.7% (85/190) from Rex rabbit. In total, ten species of Eimeria were identified from oocyst-positive samples. Concurrent infection with two to eight Eimeria species was found. E. perforans was the most prevalent species (35.2%), followed in order by E. media, E. magna, E. irresidua and E. intestinalis with prevalences of 31.3%, 28.8%, 19.4%, and 14.8%, respectively. Taken together, These results reveal the characteristics of the prevelance of rabbit coccidia infection in China, including the distribution, the scale of farming and the species, which are indispensable to the control of rabbits coccidiosis in China.

  14. Alleviation of metabolic abnormalities induced by excessive fructose administration in Wistar rats by Spirulina maxima

    PubMed Central

    Jarouliya, Urmila; Anish, Zacharia J.; Kumar, Pravin; Bisen, P.S.; Prasad, G.B.K.S.

    2012-01-01

    Background & objectives: Diabetes mellitus is a metabolic disorder characterized by hyperglycaemia. Several natural products have been isolated and identified to restore the complications of diabetes. Spirulina maxima is naturally occurring fresh water cyanobacterium, enriched with proteins and essential nutrients. The aim of the study was to determine whether S. maxima could serve as a therapeutic agent to correct metabolic abnormalities induced by excessive fructose administration in Wistar rats. Methods: Oral administration of 10 per cent fructose solution to Wistar rats (n=5 in each group) for 30 days resulted in hyperglycaemia and hyperlipidaemia. Aqueous suspension of S. maxima (5 or 10%) was also administered orally once daily for 30 days. The therapeutic potential of the preparation with reference to metformin (500 mg/kg) was assessed by monitoring various biochemical parameters at 10 day intervals during the course of therapy and at the end of 30 days S. maxima administration. Results: Significant (P<0.001) reductions in blood glucose, lipid profile (triglycerides, cholesterol and LDL, VLDL) and liver function markers (SGPT and SGOT) were recorded along with elevated level of HDL-C at the end of 30 days therapy of 5 or 10 per cent S. maxima aquous extract. Co-administration of S. maxima extract (5 or 10% aqueous) with 10 per cent fructose solution offered a significant protection against fructose induced metabolic abnormalities in Wistar rats. Interpretation & Conclusions: The present findings showed that S. maxima exhibited anti-hyperglycaemic, anti-hyperlipidaemic and hepatoprotective activity in rats fed with fructose. Further studies are needed to understand the mechanisms. PMID:22561632

  15. Accumulation of alkaline earth metals by the green macroalga Bryopsis maxima.

    PubMed

    Takahashi, Shigekazu; Aizawa, Kyoko; Nakamura, Saki; Nakayama, Katsumi; Fujisaki, Shingo; Watanabe, Soichiro; Satoh, Hiroyuki

    2015-04-01

    Twenty-five days after the disaster at the Fukushima Daiichi nuclear power plant in 2011, we collected samples of the green macroalga Bryopsis maxima from the Pacific coast of Japan. Bryopsis maxima is a unicellular, multinuclear, siphonous green macroalga. Radiation analysis revealed that B. maxima emitted remarkably high gamma radiation of (131)I, (134)Cs, (137)Cs, and (140)Ba as fission products of (235)U. Interestingly, B. maxima contained naturally occurring radionuclides derived from (226)Ra and (228)Ra. Analysis of element content revealed that B. maxima accumulates many ocean elements, especially high quantities of the alkaline earth metals Sr (15.9 g per dry-kg) and Ba (3.79 g per dry-kg), whereas Ca content (12.5 g per dry-kg) was lower than that of Sr and only 61 % of the mean content of 70 Japanese seaweed species. Time-course analysis determined the rate of radioactive (85)Sr incorporation into thalli to be approximately 0.13 g Sr per dry-kg of thallus per day. Subcellular fractionation of B. maxima cells showed that most of the (85)Sr was localized in the soluble fraction, predominantly in the vacuole or cytosol. Given that (85)Sr radioactivity was permeable through a dialysis membrane, the (85)Sr was considered to be a form of inorganic ion and/or bound with a small molecule. Precipitation analysis with sodium sulfate showed that more than 70% of the Sr did not precipitate as SrSO4, indicating that a proportion of the Sr may bind with small molecules in B. maxima.

  16. [Occurrence of coccidia from genus Eimeria in European bison in Białowieza Forest, Poland].

    PubMed

    Demiaszkiewicz, Aleksander W; Pyziel, Anna M

    2009-01-01

    Protozoa from the genus Eimeria belong to the least recognized parasites of the European bison. There are only a few data concerning the infection of the bison by these protozoa in Białowieza Forest. In the winter of 2007/2008 weighting 3 g fecal samples from the rectum of 16 shot bison were examined by flotation method. In 13 of them were found coccidian oocysts from genus Eimeria, which were determined to the species by morphometric study under the microscope. In all 13 positive samples, it is 81.2%, oocysts of Eimeria bovis were found. Intensiveness of this infection oscillated from 1 to 181 and the mean one was 38 oocysts. E. zuernii was found in 9 samples in the mean intensity 19 (1-154) oocysts. E. ellipsoidalis occurred in 3 bison with a very low intensity 1 to 3 oocysts. The most rarely occurred E. canadiensis, only in 2 examined bison in the number of 2 and 3 oocysts. All found coccidian species are the typical parasites of cattle. Białowieza Forest is a new place of occurrence of these parasites. European bison is a new host for E. zuernii.

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

    PubMed

    Sommer, C

    1998-01-01

    This study reports on morphological variability of Eimeria species, which may be given either by drawings or as quantitative data. The drawings may be used to facilitate identification by eye of 'unknown' Eimeria specimens, whereas quantitative data may serve as a reference set for identification by multivariate statistical techniques. The morphology of 810 Eimeria specimens was defined in binary (b/w) digital images by pixels of their oocyst outline. A Fourier transform of pixel positions yielded size and shape features. To classify coccidia, the quantitative data were employed in an agglomerative clustering by average linkage algorithm with equal weight assigned to size and shape. An inverse Fourier transform served to reconstruct oocyst outlines, i.e. outlines of average shape and size, from mean values of features in resulting clusters. Clusters were subsequently identified based on their average morphology by comparison with drawings of species in an earlier taxonomical work. Five hundred oocyst outlines were simulated for each cluster representing a species, and shape/size variability was presented in contour diagrams. Differences in species shapes, and correspondence in length and width, were seen after reconstruction by inverse Fourier transform and comparison with earlier studies.

  18. Characterisation of macrophage migration inhibitory factor from Eimeria species infectious to chickens.

    PubMed

    Miska, Katarzyna B; Fetterer, Raymond H; Lillehoj, Hyun S; Jenkins, Mark C; Allen, Patricia C; Harper, Susan B

    2007-02-01

    Macrophage migration inhibitory factor (MIF) was the first cytokine to be identified almost 40 years ago. Homologues of MIF have been isolated recently from invertebrates, making it an interesting molecule from an evolutionary as well as functional perspective. The present study represents the first report of MIF homologues in apicomplexan parasites, belonging to the genus Eimeria. A single full-length clone was isolated from Eimeria acervulina that shared between 35 and 38% amino acid identity with MIFs of vertebrates. A MIF cDNA from Eimeria tenella shared 64% amino acid identity with E. acervulina MIF. The mRNA expression was highest in merozoites, whereas developing oocysts and sporozoites expressed low to undetectable levels. Protein expression patterns were nearly identical to that observed by reverse transcriptase polymerase chain reaction (RT-PCR), suggesting strong developmental regulation. Immunofluorescence staining and co-localisation studies of E. acervulina merozoites indicated that MIF is distributed throughout the cytosol, and appears to be concentrated in the apical end of the parasite. The presence of MIF was detected in excretory/secretory (ES) products collected from E. acervulina merozoites, and isoelectric focusing indicated that three MIF isoforms are present in this stage. Phylogenetic analysis revealed that apicomplexan MIF sequences form a sister relationship to MIF-like molecules from Arabidopsis thaliana.

  19. Analysis of transcripts from intracellular stages of Eimeria acervulina using expressed sequence tags.

    PubMed

    Miska, K B; Fetterer, R H; Rosenberg, G H

    2008-04-01

    Coccidiosis in chickens is caused by 7 species of Eimeria. Even though coccidiosis is a complex disease that can be caused by any combination of these species, most of the molecular research concerning chicken coccidiosis has been limited to Eimeria tenella. The present study describes the first large-scale analysis of expressed sequence tags (ESTs) generated primarily from second-stage merozoites (and schizonts) of E. acervulina. In total, 1,847 ESTs were sequenced; these represent 1,026 unique sequences. Approximately half of the ESTs encode proteins of unknown function, or hypothetical proteins. Twenty-nine percent of the E. acervulina ESTs share significant sequence identity with sequences in the E. tenella genome. Additionally, EST hits seem to be much different compared with those of E. tenella. One of the differences is the very low number of ESTs that encode putative microneme proteins. This study underlines the potential differences in the molecular aspects of 2 Eimeria species that in the past were thought to be highly similar in nature.

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

    PubMed

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

    2014-03-01

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

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

    PubMed

    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

    2017-09-05

    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.

  2. Loop-mediated Isothermal Amplification (LAMP) Assays for the Species-specific Detection of Eimeria that Infect Chickens

    PubMed Central

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

    2015-01-01

    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. PMID:25741643

  3. Morphometric Identification, Gross and Histopathological Lesions of Eimeria Species in Japanese Quails (Coturnix coturnix japonica) in Zaria, Nigeria.

    PubMed

    Umar, H A; Lawal, I A; Okubanjo, O O; Wakawa, A M

    2014-01-01

    The objective of the study was to identify the species, gross and histopathological lesions of Eimeria in Japanese quails in Zaria. A total of 400 fresh faecal samples were collected and 10 quail birds were purchased from a quail farm. The faecal samples were processed using simple floatation technique. Oocysts shape indices of sporulated oocysts were determined. The intestines were observed for gross lesions and segments were analyzed using Giemsa stain and Haematoxylin and Eosin stain and then observed microscopically for the developmental stages of the parasite. Four species of Eimeria were identified in the study. Eimeria bateri of shape index of 1.36 conformed to the guidelines used while the other three Eimeria species with shape indices of 1.48, 1.03, and 1.40 were not confirmed. The main gross lesion seen was nonhaemorrhagic ballooning of the caeca. Intestinal scrapping smear revealed a developmental stage of the parasite (merozoites) in the jejunum. Histopathology also revealed a developmental stage (schizont) of the parasite in the caecum and desquamation of the epithelial lining with areas of necrosis. Further study is required using molecular techniques to properly identify the unknown species of Eimeria that were detected in the study.

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

    PubMed

    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.

  5. Characterization of the antibody response in birds following infection with wild-type and attenuated strains of Eimeria tenella and Eimeria necatrix.

    PubMed

    Constantinoiu, C C; Molloy, J B; Jorgensen, W K; Coleman, G T

    2011-01-10

    Live vaccines containing attenuated parasite strains are increasingly used to control chicken coccidiosis. In this paper antibody responses elicited by infections with wild-type and attenuated strains of Eimeria tenella and Eimeria necatrix were characterized by immunoblotting and ELISA with homologous and heterologous antisera. Few differences between antisera from birds infected with wild and attenuated strains of E. tenella were evident in immunoblots conducted with merozoite antigen preparations from both E. tenella strains, however the reactivity of sera raised in birds infected with the wild-type strain was noticeably more intense. In ELISAs conducted with merozoite antigen preparations, antisera from birds infected with the wild-type strains of E. tenella and E. necatrix consistently produced a significantly higher (P<0.05) antibody response than antisera from birds infected with the attenuated strains. Likewise, avidity ELISAs conducted with the E. tenella strains demonstrated that antibodies in birds infected with the wild-type strain were of significantly higher avidity (P<0.05) than antibodies in birds infected with the attenuated strain. The differences in the antibody responses are probably due to changes in the attenuated strain as a result of selection for precocious development and the less severe tissue damage and inflammation of the intestine resulting from infection with the attenuated strain.

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

    PubMed Central

    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 P<0.05−0.01 (250 spots), P<0.01−0.001 (248 spots), and P<0.001 (314 spots) were excised and analyzed by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry. Proteins were identified in 172 spots. A total of 46 different proteins were identified. Of the spots with a corresponding protein identification, 57 showed a main effect of coccidia infection and/or 2-way interaction of coccidia infection×broiler genetic line at P<0.001. Conclusions/Significance Several of the metabolic enzymes identified in this study are potential candidates for early diagnostic markers of E. acervulina infection including malate dehydrogenase 2, NADH dehydrogenase 1 alpha subcomplex 9, and an ATP synthase. These proteins were detected only in Line A birds that were inoculated with E. acervulina. Results from this study provide a basic framework for future research aimed at uncovering the complex

  7. Eimeria species and genetic background influence the serum protein profile of broilers with coccidiosis.

    PubMed

    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

    2011-01-31

    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. 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 P<0.05-0.01 (250 spots), P<0.01-0.001 (248 spots), and P<0.001 (314 spots) were excised and analyzed by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry. Proteins were identified in 172 spots. A total of 46 different proteins were identified. Of the spots with a corresponding protein identification, 57 showed a main effect of coccidia infection and/or 2-way interaction of coccidia infection×broiler genetic line at P<0.001. Several of the metabolic enzymes identified in this study are potential candidates for early diagnostic markers of E. acervulina infection including malate dehydrogenase 2, NADH dehydrogenase 1 alpha subcomplex 9, and an ATP synthase. These proteins were detected only in Line A birds that were inoculated with E. acervulina. Results from this study provide a basic framework for future research aimed at uncovering the complex biochemical mechanisms involved in host response to Eimeria infection

  8. Mutational analysis of the pumpkin (Cucurbita maxima) phloem exudate lectin, PP2 reveals Ser-104 is crucial for carbohydrate binding.

    PubMed

    Bobbili, Kishore Babu; Bandari, Shyam; Grobe, Kay; Swamy, Musti J

    2014-07-18

    The pumpkin phloem lectin (PP2) is an RNA-binding, defense-related, chitooligosaccharide-specific, homodimeric lectin of Mr 48 kDa expressed at high concentrations in the sieve elements and companion cells of pumpkin (Cucurbita maxima). In the present study, PP2 was expressed in the methylotrophic yeast Pichia pastoris with the Saccharomyces α-factor sequence to direct the recombinant protein into the secretory pathway as a prerequisite for unimpaired folding and posttranslational glycosylation of recombinant PP2. Previous computational modeling and ligand docking studies predicted a putative chitooligosaccharide-binding site on the PP2 surface, which was divided into three subsites, with two amino acid residues in each subsite identified as possible candidates for interaction with chitooligosaccharides (CHOs). In this work, mutational analysis and hemagglutination assays were employed to verify the role of the predicted residues in the carbohydrate binding activity of the protein. The results obtained revealed that mutation of Ser-104 to Ala (S104A) at subsite-2 resulted in about 90% loss of agglutination activity of the protein, indicating that Ser-104 is crucial for the binding of CHOs to PP2. Also, L100A (at subsite-1) and K200A (at subsite-3) independently decreased the lectin activity by about 40%, indicating that these two residues also contribute significantly to sugar binding by PP2. Together, these findings confirm that all the three subsites contribute to varying degrees toward PP2-carbohydrate interaction, and confirm the validity of the computational model, as proposed earlier. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. Extreme value theory (EVT) application on estimating the distribution of maxima

    NASA Astrophysics Data System (ADS)

    Ramadhani, F. A.; Nurrohmah, S.; Novita, M.

    2017-07-01

    Extreme Value Theory (EVT) has emerged as one of the most important statistical theories for the applied sciences. EVT provides a firm theoretical foundation for building a statistical model describing extreme events. The feature that distinguish extreme value analysis than other statistical analysis is the ability to quantify the behavior of unusually large values even when those values are scarce. One of the key results from EVT is the ability to estimate the distribution of maximum value, that usually called as maxima, using the asymptotic argument. In order to build such models, the Fisher-Tippett theorem which specifies the form of the limit distribution for transformed maxima will be greatly used. Furthermore, it can be shown that there are only three families of possible limit laws for distribution of maxima, which are the Gumbel, Frechet, and Weibull distributions. These three distributions can be expressed in a single distribution function called the generalized extreme value (GEV) distribution.

  10. Use of extracts from oyster shell and soil for cultivation of Spirulina maxima.

    PubMed

    Jung, Joo-Young; Kim, Sunmin; Lee, Hansol; Kim, Kyochan; Kim, Woong; Park, Min S; Kwon, Jong-Hee; Yang, Ji-Won

    2014-12-01

    Calcium ion and trace metals play important roles in various metabolisms of photosynthetic organisms. In this study, simple methods were developed to extract calcium ion and micronutrients from oyster shell and common soil, and the prepared extracts were tested as a replacement of the corresponding chemicals that are essential for growth of microalgae. The oyster shell and soil were treated with 0.1 M sodium hydroxide or with 10 % hydrogen peroxide, respectively. The potential application of these natural sources to cultivation was investigated with Spirulina maxima. When compared to standard Zarrouk medium, the Spirulina maxima cultivated in a modified Zarrouk media with elements from oyster shell and soil extract exhibited increases in biomass, chlorophyll, and phycocyanin by 17, 16, and 64 %, respectively. These results indicate that the extracts of oyster shell and soil provide sufficient amounts of calcium and trace metals for successful cultivation of Spirulina maxima.

  11. Time spans between price maxima and price minima in stock markets

    NASA Astrophysics Data System (ADS)

    Zou, Yongjie; Li, Honggang

    2014-02-01

    We empirically investigate the distribution of time spans between price maxima and price minima in international stock markets, where a time span is defined as the time interval between a local price minimum and a local price maximum, and local price extrema are identified by a method introduced by Preis and Stanley (Preis et al. (2011), Preis (2011), Preis and Stanley (2011, 2010), Preis (2010), Preis and Stanley (2010), Stanley et al. (2010), Preis and Stanley (2009)). The empirical results show that both the tail distributions of time spans from local price maxima to local price minima and the tail distributions of time spans from local price minima to local price maxima yield an exponential distribution. In addition, price rise/fall asymmetry is observed by comparing the values of the exponents of the distribution curves. These results are robust across eight representative stock markets.

  12. Evaluation of Antidepressant-like Effect of Citrus Maxima Leaves in Animal Models of Depression

    PubMed Central

    Potdar, Vikram H; Kibile, Swati J

    2011-01-01

    Objective(s) This study planned to assess antidepressant like activity of aqueous extract from leaves of Citrus maxima Merr. (Rutaceae). Materials and Methods Boiling was used for aqueous extraction. Acute toxicity study was performed in mice. Antidepressant activity was studied using locomotor activity test, modified forced swimming test (FST) and tail suspension test (TST). Three doses 100, 200 and 300 mg/kg of aqueous extract of leaves were selected for testing. Fluoxetine (20 mg/kg, i.p.) and imipramine (30 mg/kg, i.p.) were used as the standard drugs. Results Aqueous extract of Citrus maxima leaves significantly reduced immobility time in both TST and FST. In locomotor activity testing it showed psychostimulant effect. Extract increased the climbing behavior in FST, which is similar to effect observed with imipramine. Conclusion The results of this study suggest that antidepressant like effect of Citrus maxima seems to be mediated by an increase in norepinephrine level in synapses. PMID:23492865

  13. Maxima of |Ψ|2: a connection between quantum mechanics and Lewis structures.

    PubMed

    Lüchow, Arne

    2014-04-30

    The maxima of squared electronic wave functions |Ψ|2 are analyzed for a number of small molecules. They are in principle observables and show considerable chemical insight from first principles. The maxima contain substantial information about the relative electron positions in a molecule, such as the pairing of opposite spin electrons and the Pauli repulsion which are lost in the electron density. Single bond and double bond as well as polar bond pairs and lone pairs are obtained from the maximum analysis. In many cases, we find a correspondence to the electron arrangements in molecules as assumed by Lewis in 1916.

  14. VizieR Online Data Catalog: Minima and maxima of 389 variables (Hubscher+, 2007)

    NASA Astrophysics Data System (ADS)

    Hubscher, J.; Walter, F.

    2008-11-01

    In this 57th compilation of BAV results, photoelectric observations obtained in the year 2006 are presented on 389 variable stars giving 611 minima on eclipsing binaries and maxima on pulsating stars. All moments of minima and maxima are heliocentric. The errors are tabulated in column '+/-'. The values in column 'O-C' are determined without incorporation of nonlinear terms. The references are given in the section 'Remarks'. All information about photometers and filters are specified in the column 'Rem'. The observations were made at private observatories. The photoelectric measurements and all the light-curves with evaluations can be obtained from the office of the BAV for inspection. (3 data files).

  15. VizieR Online Data Catalog: Minima and maxima of 380 variables (Hubscher+, 2009)

    NASA Astrophysics Data System (ADS)

    Hubscher, J.; Steinbach, H.-M.; Walter, F.

    2011-03-01

    In this 63rd compilation of BAV results, photoelectric observations obtained in the year 2008 are presented on 380 variable stars giving 591 minima on eclipsing binaries and maxima on pulsating stars. All moments of minima and maxima are heliocentric. The errors are tabulated in column '+/-'. The values in column 'O-C' are determined without incorporation of nonlinear terms. The references are given in the section 'Remarks'. All information about photometers and filters are specified in the column 'Rem'. The observations were made at private observatories. The photoelectric measurements and all the light curves with evaluations can be obtained from the office of the BAV for inspection. (3 data files).

  16. VizieR Online Data Catalog: Minima and maxima of 436 variables (Huebscher+, 2011)

    NASA Astrophysics Data System (ADS)

    Huebscher, J.; Monninger, G.

    2011-08-01

    In this 68th compilation of BAV results, photoelectric observations obtained in the year 2010 are presented on 436 variable stars giving 784 minima on eclipsing binaries and maxima on pulsating stars. All moments of minima and maxima are heliocentric. The errors are tabulated in column '+/-'. The values in column 'O-C' are determined without incorporation of nonlinear terms.The references are given in the section 'Remarks'. All information about photometers and filters are specified in the column 'Rem'. The observations were made at private observatories. The photoelectric measurements and all the lightcurves with evaluations can be obtained from the office of the BAV for inspection. (3 data files).

  17. VizieR Online Data Catalog: Minima and maxima of 292 variables (Hubscher+, 2008)

    NASA Astrophysics Data System (ADS)

    Hubscher, J.; Steinbach, H.-M.; Walter, F.

    2008-11-01

    In this 60th compilation of BAV results, photoelectric observations obtained in the years 2007 are presented on 292 variable stars giving 399 minima on eclipsing binaries and maxima on pulsating stars. All moments of minima and maxima are heliocentric. The errors are tabulated in column '+/-'. The values in column 'O-C' are determined without incorporation of nonlinear terms. The references are given in the section 'Remarks'. All information about photometers and filters are specified in the column 'Rem'. The observations were made at private observatories. The photoelectric measurements and all the lightcurves with evaluations can be obtained from the office of the BAV for inspection. (3 data files).

  18. VizieR Online Data Catalog: Minima and maxima of 452 variables (Hubscher+, 2010)

    NASA Astrophysics Data System (ADS)

    Hubscher, J.; Lehmann, P. B.; Monninger, G.; Steinbach, H.-M.; Walter, F.

    2011-03-01

    In this 65th compilation of BAV results, photoelectric observations obtained in the year 2009 are presented on 521 variable stars giving 871 minima on eclipsing binaries and maxima on pulsating stars. All moments of minima and maxima are heliocentric. The errors are tabulated in column '+/-'. The values in column 'O-C' are determined without incorporation of nonlinear terms. The references are given in the section 'Remarks'. All information about photometers and filters are specified in the column 'Rem'. The observations were made at private observatories. The photoelectric measurements and all the light curves with evaluations can be obtained from the office of the BAV for inspection. (3 data files).

  19. VizieR Online Data Catalog: Minima and maxima of 503 variables (Hubscher, 2011)

    NASA Astrophysics Data System (ADS)

    Hubscher, J.

    2011-09-01

    In this 69th compilation of BAV results, photoelectric observations obtained in the year 2010 and 2011 are presented on 503 variable stars giving 767 minima on eclipsing binaries and maxima on pulsating stars. All moments of minima and maxima are heliocentric. The errors are tabulated in column '+/-'. The values in column 'O-C' are determined without incorporation of nonlinear terms. The references are given in the section 'Remarks'. All information about photometers and filters are specified in the column 'Rem'. The observations were made at private observatories. The photoelectric measurements and all the light curves with evaluations can be obtained from the office of the BAV for inspection. (3 data files).

  20. VizieR Online Data Catalog: Minima and maxima of 735 variables (Hubscher+, 2007)

    NASA Astrophysics Data System (ADS)

    Hubscher, J.

    2008-11-01

    In this 58th compilation of BAV results, photoelectric observations obtained in the years 2006 and 2007 are presented on 473 variable stars giving 735 minima and maxima on eclipsing binaries and pulsating stars. All moments of minima and maxima are heliocentric. The errors are tabulated in column '+/-'. The values in column 'O-C' are determined without incorporation of nonlinear terms. The references are given in the section 'Remarks'. All information about photometers and filters are specified in the column 'Rem'. The observations were made at private observatories. The photoelectric measurements and all the light curves with evaluations can be obtained from the office of the BAV for inspection. (3 data files).

  1. Sub-daily precipitation disaggregation for a simulation of annual runoff maxima

    NASA Astrophysics Data System (ADS)

    Sikorska, Anna; Seibert, Jan

    2017-04-01

    Measurement of precipitation has been often restricted to daily totals only. This applies especially to observations from the previous century which constitutes a valuable source of long-term precipitation records needed for statistical analysis such as for annual runoff maxima. In this respect, the knowledge about the exact temporal distribution of these precipitation totals at sub-daily time steps can play an important role on the magnitude of simulated runoff maxima using these data. But it is difficult to estimate the sub-daily temporal distribution of precipitation if it was not measured. Instead, the effect of the precipitation in the form of runoff measured at the catchment outlet, which is usually available at sub-daily time steps for longer periods, provides an alternative to evaluate the potential effect of the need to disaggregate precipitation data. In this study we assess how the choice of the temporal distribution of daily precipitation totals might affect the simulation of the catchment runoff annual maxima when using a hydrological model. To examine this issue, we tested six different settings of precipitation total distribution within the day. These are: 1) uniform distribution (daily totals uniquely divided over 24 hours); all daily totals fall within a respective time window, i.e., 2) one hour, 3) two successive hours, 4) three successive hours, 5) six successive hours, and 6) twelve successive hours, each time randomly selected for each observation day independently. To assess the effect on simulated runoff maxima, such generated hourly precipitation datasets were next used as input into a pre-calibrated HBV model. As a reference, we used model simulations with observed hourly precipitation data. This study was conducted using thirty years of precipitation and runoff observations in three Swiss catchments. Our results showed that the annual maxima were best simulated when distributing daily totals over twelve successive hours randomly selected

  2. Joint distributions of partial and global maxima of a Brownian bridge

    NASA Astrophysics Data System (ADS)

    Bénichou, Olivier; Krapivsky, P. L.; Mejía-Monasterio, Carlos; Oshanin, Gleb

    2016-08-01

    We analyze the joint distributions and temporal correlations between the partial maximum m and the global maximum M achieved by a Brownian bridge on the subinterval [0,{t}1] and on the entire interval [0,t], respectively. We determine three probability distribution functions: the joint distribution P(m,M) of both maxima; the distribution P(m) of the partial maximum; and the distribution {{\\Pi }}(G) of the gap between the maxima, G=M-m. We present exact results for the moments of these distributions and quantify the temporal correlations between m and M by calculating the Pearson correlation coefficient.

  3. High-throughput capillary electrophoresis for the identification and differentiation of seven species of Eimeria from chickens.

    PubMed

    Gasser, Robin B; Skinner, Rob; Fadavi, Reza; Richards, Grant; Morris, Genevieve

    2005-09-01

    A capillary electrophoretic approach has been evaluated for the identification of seven currently recognised species of Eimeria infecting chickens. The second internal transcribed spacer of ribosomal DNA is PCR-amplified from any of the seven species using a single set of oligonucleotide primers (one of which is fluorescently labelled). The amplicons are heat-denatured and subjected to capillary electrophoresis in a MegaBACE 1000 (Amersham). The chromatograms captured are stored electronically and then analysed using MegaBACE Fragment Profiler software. Using control DNA samples representing monospecific lines of Eimeria, specific peaks in the chromatograms were defined for the unequivocal identification of each of the seven species and their differentiation. Electrophoretic reading and analysis are carried out automatically, thus making it a time- and cost-effective method. This procedure should find applicability as a tool for the quality control of Eimeria vaccines, the monitoring of coccidiosis outbreaks and the high-throughput analysis of oocyst samples for epidemiological surveys.

  4. Control of coccidiosis in turkeys with diclazuril and monensin: effects upon performance and development of immunity to Eimeria species.

    PubMed

    Chapman, H D; Matsler, P L; Chapman, M E

    2004-09-01

    The effects of diclazuril and monensin, when included in the feed of turkeys from 0 to 10 wk, upon performance and development of immunity to Eimeria species was investigated. Birds were initially inoculated with a low dose of oocysts of three species of Eimeria at 3, 5, 7, and 9 days of age in order to simulate a natural infection. Weight gain and feed intake from 0 to 6 wk of age was significantly greater in medicated birds compared with those that received no anticoccidial medication. Weight gain and feed intake from 6 to 10 wk was greater in birds that received diclazuril than in unmedicated birds. No differences in performance were evident after drug withdrawal from 10 to 16 wk. Immunity to Eimeria species developed by 10 wk in birds that received no anticoccidial medication but did not develop in those given diclazuril or monensin.

  5. The effect of mitochondrial ATP-sensitive potassium channels on apoptosis of chick embryo cecal cells by Eimeria tenella.

    PubMed

    Yang, Sha-sha; Zheng, Ming-xue; Xu, Huan-cheng; Cui, Xiao-zhen; Zhang, Yan; Zhao, Wen-long; Bai, Rui

    2015-04-01

    The objective of this study was to investigate the effect of mitochondrial ATP-sensitive potassium (mitoKATP) channels on apoptosis induced by Eimeria tenella. At 24, 48, 72, 96 and 120 h after Eimeria tenella infection, TUNEL assays and translation of phosphatidyl serines to the host cell plasma membrane surface showed that diazoxide-treated chick embryo cecal cells underwent less apoptosis (P <0.05), while light microscopy showed that infection rates of treated cells were higher (P <0.01) than untreated cells. Caspase 9 and caspase 3 of infected cells were activated less (P <0.01) in diazoxide-treated cells than untreated cells. These results indicate that opening mitoKATP channels can protect chick embryo cecal cells from mitochondria-dependent apoptosis induced by Eimeria tenella by inhibiting activations of caspase 9 and caspase 3.

  6. Evidence for a Structural Role for Acid-Fast Lipids in Oocyst Walls of Cryptosporidium, Toxoplasma, and Eimeria

    PubMed Central

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

    2013-01-01

    ABSTRACT 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). PMID:24003177

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

    PubMed

    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.

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

    PubMed

    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.

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

    PubMed

    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.

  10. Antioxidant Effect of Spirulina (Arthrospira) maxima on Chronic Inflammation Induced by Freund's Complete Adjuvant in Rats.

    PubMed

    Gutiérrez-Rebolledo, Gabriel Alfonso; Galar-Martínez, Marcela; García-Rodríguez, Rosa Virginia; Chamorro-Cevallos, Germán A; Hernández-Reyes, Ana Gabriela; Martínez-Galero, Elizdath

    2015-08-01

    One of the major mechanisms in the pathogenesis of chronic inflammation is the excessive production of reactive oxygen and reactive nitrogen species, and therefore, oxidative stress. Spirulina (Arthrospira) maxima has marked antioxidant activity in vivo and in vitro, as well as anti-inflammatory activity in certain experimental models, the latter activity being mediated probably by the antioxidant activity of this cyanobacterium. In the present study, chronic inflammation was induced through injection of Freund's complete adjuvant (CFA) in rats treated daily with Spirulina (Arthrospira) maxima for 2 weeks beginning on day 14. Joint diameter, body temperature, and motor capacity were assessed each week. On days 0 and 28, total and differential leukocyte counts and serum oxidative damage were determined, the latter by assessing lipid peroxidation and protein carbonyl content. At the end of the study, oxidative damage to joints was likewise evaluated. Results show that S. maxima favors increased mobility, as well as body temperature regulation, and a number of circulating leukocytes, lymphocytes, and monocytes in specimens with CFA-induced chronic inflammation and also protects against oxidative damage in joint tissue as well as serum. In conclusion, the protection afforded by S. maxima against development of chronic inflammation is due to its antioxidant activity.

  11. Antioxidant Effect of Spirulina (Arthrospira) maxima on Chronic Inflammation Induced by Freund's Complete Adjuvant in Rats

    PubMed Central

    Gutiérrez-Rebolledo, Gabriel Alfonso; Galar-Martínez, Marcela; García-Rodríguez, Rosa Virginia; Chamorro-Cevallos, Germán A.; Hernández-Reyes, Ana Gabriela

    2015-01-01

    Abstract One of the major mechanisms in the pathogenesis of chronic inflammation is the excessive production of reactive oxygen and reactive nitrogen species, and therefore, oxidative stress. Spirulina (Arthrospira) maxima has marked antioxidant activity in vivo and in vitro, as well as anti-inflammatory activity in certain experimental models, the latter activity being mediated probably by the antioxidant activity of this cyanobacterium. In the present study, chronic inflammation was induced through injection of Freund's complete adjuvant (CFA) in rats treated daily with Spirulina (Arthrospira) maxima for 2 weeks beginning on day 14. Joint diameter, body temperature, and motor capacity were assessed each week. On days 0 and 28, total and differential leukocyte counts and serum oxidative damage were determined, the latter by assessing lipid peroxidation and protein carbonyl content. At the end of the study, oxidative damage to joints was likewise evaluated. Results show that S. maxima favors increased mobility, as well as body temperature regulation, and a number of circulating leukocytes, lymphocytes, and monocytes in specimens with CFA-induced chronic inflammation and also protects against oxidative damage in joint tissue as well as serum. In conclusion, the protection afforded by S. maxima against development of chronic inflammation is due to its antioxidant activity. PMID:25599112

  12. Removal of local and biased global maxima in intensity-based registration.

    PubMed

    Salvado, Olivier; Wilson, David L

    2007-04-01

    Intensity based registration (e.g., mutual information) suffers from a scalloping artifact giving rise to local maxima and sometimes a biased global maximum in a similarity objective function. Here, we demonstrate that scalloping is principally due to the noise reduction filtering that occurs when image samples are interpolated. Typically at a much smaller scale (100 times less in our test cases), there are also fluctuations in the similarity objective function due to interpolation of the signal and to sampling of a continuous, band-limited image signal. Focusing on the larger problem from noise, we show that this phenomenon can even bias global maxima, giving inaccurate registrations. This phenomenon is readily seen when one registers an image onto itself with different noise realizations but is absent when the same noise realization is present in both images. For linear interpolation, local maxima and global bias are removed if one filters the interpolated image using a new constant variance filter for linear interpolation (cv-lin filter), which equalizes the variance across the interpolated image. We use 2D synthetic and MR images and characterize the effect of cv-lin on similarity objective functions. With a reduction of local and biased maxima, image registration becomes more robust and accurate. An efficient implementation adds insignificant computation time per iteration, and because optimization proceeds more smoothly, sometimes fewer iterations are needed.

  13. Potential antiradical and alpha-glucosidase inhibitors from Ecklonia maxima (Osbeck) Papenfuss.

    PubMed

    Rengasamy, Kannan R R; Aderogba, Mutalib A; Amoo, Stephen O; Stirk, Wendy A; Van Staden, Johannes

    2013-11-15

    Alpha-glucosidase inhibitors play a potential role in the treatment of type 2 diabetes by delaying glucose absorption in the small intestine. Ecklonia maxima, a brown alga which grows abundantly on the west coast of South Africa, is used to produce alginate, animal feed, nutritional supplements and fertilizer. The crude aqueous methanol extract, four solvent fractions and three phlorotannins: 1,3,5-trihydroxybenezene (phloroglucinol) (1), dibenzo [1,4] dioxine-2,4,7,9-tetraol (2) and hexahydroxyphenoxydibenzo [1,4] dioxine (eckol) (3) isolated from E. maxima were evaluated for antiradical and alpha-glucosidase inhibitory activities. All the phlorotannins tested had strong antioxidant activities on DPPH free radicals with EC50 values ranging from 0.008 to 0.128μM. Compounds 2 and 3 demonstrated stronger antioxidant activity and an alpha-glucosidase inhibitory property than positive controls. These results suggest that E. maxima could be a natural source of potent antioxidants and alpha-glucosidase inhibitors. This study could facilitate effective utilization of E. maxima as an oral antidiabetic drug or functional food ingredient with a promising role in the formulation of medicines and nutrition supplements.

  14. Antioxidative activity of water soluble polysaccharide in pumpkin fruits (Cucurbita maxima Duchesne).

    PubMed

    Nara, Kazuhiro; Yamaguchi, Akira; Maeda, Naomi; Koga, Hidenori

    2009-06-01

    We evaluated the antioxidative activity of a water soluble polysaccharide fraction (WSP) from pumpkin fruits (Cucurbita maxima Duchesne). In the WSP, DPPH radical scavenging and superoxide dismutase-like activity increased depending on the total sugar content. Furthermore, the WSP can serve as an inhibitor of ascorbic acid oxidation. The efficacy was also affected by the total sugar content.

  15. An Example of Competence-Based Learning: Use of Maxima in Linear Algebra for Engineers

    ERIC Educational Resources Information Center

    Diaz, Ana; Garcia, Alfonsa; de la Villa, Agustin

    2011-01-01

    This paper analyses the role of Computer Algebra Systems (CAS) in a model of learning based on competences. The proposal is an e-learning model Linear Algebra course for Engineering, which includes the use of a CAS (Maxima) and focuses on problem solving. A reference model has been taken from the Spanish Open University. The proper use of CAS is…

  16. An Example of Competence-Based Learning: Use of Maxima in Linear Algebra for Engineers

    ERIC Educational Resources Information Center

    Diaz, Ana; Garcia, Alfonsa; de la Villa, Agustin

    2011-01-01

    This paper analyses the role of Computer Algebra Systems (CAS) in a model of learning based on competences. The proposal is an e-learning model Linear Algebra course for Engineering, which includes the use of a CAS (Maxima) and focuses on problem solving. A reference model has been taken from the Spanish Open University. The proper use of CAS is…

  17. Beyond Polymaxenolide: Cembrane-Africanane Terpenoids from the Hybrid Soft Coral Sinularia maxima x S. polydactyla

    USDA-ARS?s Scientific Manuscript database

    The effects of natural hybridization on secondary metabolite production and diversification have only recently been studied in plants and have essentially been overlooked in marine organisms. Chemical investigation of the hybrid soft coral Sinularia maxima × S. polydactyla resulted in the isolation ...

  18. Phloem-associated auxin response maxima determine radial positioning of lateral roots in maize

    PubMed Central

    Jansen, Leentje; Roberts, Ianto; De Rycke, Riet; Beeckman, Tom

    2012-01-01

    In Arabidopsis thaliana, lateral-root-forming competence of pericycle cells is associated with their position at the xylem poles and depends on the establishment of protoxylem-localized auxin response maxima. In maize, our histological analyses revealed an interruption of the pericycle at the xylem poles, and confirmed the earlier reported proto-phloem-specific lateral root initiation. Phloem-pole pericycle cells were larger and had thinner cell walls compared with the other pericycle cells, highlighting the heterogeneous character of the maize root pericycle. A maize DR5::RFP marker line demonstrated the presence of auxin response maxima in differentiating xylem cells at the root tip and in cells surrounding the proto-phloem vessels. Chemical inhibition of auxin transport indicated that the establishment of the phloem-localized auxin response maxima is crucial for lateral root formation in maize, because in their absence, random divisions of pericycle and endodermis cells occurred, not resulting in organogenesis. These data hint at an evolutionarily conserved mechanism, in which the establishment of vascular auxin response maxima is required to trigger cells in the flanking outer tissue layer for lateral root initiation. It further indicates that lateral root initiation is not dependent on cellular specification or differentiation of the type of vascular tissue. PMID:22527395

  19. Phloem-associated auxin response maxima determine radial positioning of lateral roots in maize.

    PubMed

    Jansen, Leentje; Roberts, Ianto; De Rycke, Riet; Beeckman, Tom

    2012-06-05

    In Arabidopsis thaliana, lateral-root-forming competence of pericycle cells is associated with their position at the xylem poles and depends on the establishment of protoxylem-localized auxin response maxima. In maize, our histological analyses revealed an interruption of the pericycle at the xylem poles, and confirmed the earlier reported proto-phloem-specific lateral root initiation. Phloem-pole pericycle cells were larger and had thinner cell walls compared with the other pericycle cells, highlighting the heterogeneous character of the maize root pericycle. A maize DR5::RFP marker line demonstrated the presence of auxin response maxima in differentiating xylem cells at the root tip and in cells surrounding the proto-phloem vessels. Chemical inhibition of auxin transport indicated that the establishment of the phloem-localized auxin response maxima is crucial for lateral root formation in maize, because in their absence, random divisions of pericycle and endodermis cells occurred, not resulting in organogenesis. These data hint at an evolutionarily conserved mechanism, in which the establishment of vascular auxin response maxima is required to trigger cells in the flanking outer tissue layer for lateral root initiation. It further indicates that lateral root initiation is not dependent on cellular specification or differentiation of the type of vascular tissue.

  20. The role of wind in generating turbidity maxima in the Tay Estuary

    NASA Astrophysics Data System (ADS)

    Weir, D. J.; McManus, J.

    1987-11-01

    Variations of wind direction and strength in the Tay Estuary control wave generation and the resultant patterns of suspended sediment concentration in the waters over the extensive tidal flats. Bodies of water with high water concentration peaks advect to occupy sites at which turbidity maxima are normally present at low water. Other low water peaks are generated by ebb tidal current resuspension.

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

    PubMed

    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.

  2. Protection Efficacy of Multivalent Egg Yolk Immunoglobulin against Eimeria tenella Infection in Chickens

    PubMed Central

    Xu, JJ; Ren, CZ; Wang, SS; Liu, DD; Cao, LQ; Tao, JP

    2013-01-01

    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 (P<0.05). No significant difference was observed at dosage ≥ 0.5% and 1.0% when BWG and caecal lesion were compared with the sodium salinomycin control group, respectively (P>0.05). Conclusion Supplementing newly hatched chicks with Eimeria-specific IgY represents a promising strategy to prevent avian coccidiosis. PMID:24454440

  3. Eimeria bovis-induced modulation of the host cell proteome at the meront I stage.

    PubMed

    Lutz, Kathleen; Schmitt, Sigrid; Linder, Monica; Hermosilla, Carlos; Zahner, Horst; Taubert, Anja

    2011-01-01

    The proteome of Eimeria bovis meront I-carrying host cells was analyzed by two-dimensional gel electrophoresis (2DE) at 14 days p.i. and compared to non-infected control cells. A total of 221 protein spots were modulated in their abundance in E. bovis-infected host cells and were subsequently analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectometry (MALDI-TOF-MS). These analyses identified 104 proteins in total with 25 host cell proteins being up-regulated and 79 proteins being down-regulated in E. bovis-infected host cells. Moreover, 20 newly expressed proteins were identified exclusively in E. bovis-infected host cells and were most likely of parasite origin. Parasite-induced differences in protein abundance concerned distinct functional categories, with most proteins being involved in host cell metabolism, cell structure, protein fate and gene transcription. Some of the modulated molecules also indicated regulatory processes on the level of host cell stress response (HSP70, HSP90), host cell apoptosis (caspase 8) and actin elongation/depolymerization (α-actinin-1, gelsonin, tropomodulin-3, transgelin). Since merozoites I were already released shortly after cell sampling, the current data reflect the situation at the end of first merogony. This is the first proteomic approach on E. bovis-infected host cells that was undertaken to gain a rather broad insight into Eimeria-induced host cell modulation. The data processed in this investigation should provide a useful basis for more detailed analyses concerning Eimeria-host cell interactions.

  4. Clinical and epidemiological characteristics of Eimeria infections in first-year grazing cattle.

    PubMed

    von Samson-Himmelstjerna, G; Epe, C; Wirtherle, N; von der Heyden, V; Welz, C; Radeloff, I; Beening, J; Carr, D; Hellmann, K; Schnieder, T; Krieger, K

    2006-03-31

    Infections with Eimeria parasites can lead to severe diarrhoea with considerable clinical and economic consequences in first-year grazing stock. To identify and characterise the cause of diarrhoea observed during previous years, 164 animals on 14 dairy farms in northwestern Germany were included in this study. The calves were physically and parasitologically examined prior to turnout and until 21 days post turnout (d.p.t.). Mean animal weights decreased from 194.9 kg at the start to 189.3 kg bodyweight at the end of the study. In all herds, oocyst counts were very low prior to turnout and increased after the calves had been kept on pasture for at least 7 days. On Day 9 post turnout, 90% and at the end of the study (21 d.p.t.) 70% of all animals showed Eimeria-positive faecal samples. During the course of the study, 79 (48.2%) animals passed faecal samples with more than 100,000 oocysts per gram. The predominant species identified was Eimeria alabamensis, which accounted for more than 83% of the oocysts counted. These parasitological findings matched the clinical observations. Diarrhoea was found in 130 (79.3%) of the study animals. At 5d.p.t. and thus prior to the rise of faecal oocyst counts, a significant increase in diarrhoea was recorded. Calves showing diarrhoea excreted statistically significantly more often over 100,000 E. alabamensis oocysts per gram faeces (0.28; p = 0.0002) than calves without diarrhoea. Diarrhoea was also found during significantly more study days in animals with high oocyst counts (0.39; p = 0.0001). These data indicate that in endemic areas first-year grazing calves must be considered at risk to develop clinical coccidiosis due to E. alabamensis infection during the first 2-3 weeks post turnout.

  5. Efficacy of toltrazuril (Baycox 5% suspension) in natural infections with pathogenic Eimeria spp. in housed lambs.

    PubMed

    Le Sueur, C; Mage, C; Mundt, H-C

    2009-04-01

    A field study was conducted to evaluate the effect of a single oral treatment with 20 mg/kg body weight (BW) of toltrazuril (Baycox 5% suspension)--TOL--in comparison to a single oral treatment with 1 mg/kg BW of diclazuril (Vecoxan suspension orale, 2.5 mg/ml)--DIC--and an untreated control group (CTRL) on naturally acquired Eimeria infections in lambs. On a French sheep farm with a known history of coccidiosis, 75 housed lambs aged 10-14 days were randomised and allocated to one of three groups. During an observation period of 60 days after treatment, clinical (faecal consistency, BW) and parasitological parameters (oocyst excretion) were evaluated. Excretion in the negative control group started 3 days after treatment and peaked on the 31st day with a prevalence of 80%. Animals were predominantly infected with Eimeria ovinoidalis. Treatment with toltrazuril, but not with diclazuril, resulted in significantly reduced numbers of excreting animals. The number of excretion days and the average oocyst excretion decreased significantly in both the TOL and the DIC groups compared to the CTRL, with the TOL group showing significantly fewer excretion days and excretion intensities than the DIC group. Changes in the faecal consistency were moderate throughout the study and not significantly different between the groups. Daily weight gains were higher in the TOL group compared to the DIC and CTRL groups which did not differ. This study demonstrates the good efficacy of toltrazuril administered orally to lambs in the prepatent period in subclinical natural Eimeria infections in housed lambs.

  6. Eimeria tenella oocyst shedding and output in cecal or fecal contents following experimental challenge in broilers.

    PubMed

    Jordan, A; Caldwell, D J; Klein, J; Coppedge, J; Pohl, S; Fitz-Coy, S; Lee, J T

    2011-05-01

    Two experiments were conducted to determine whether Eimeria tenella oocyst output in cecal and fecal contents, lesion development, and performance characteristics were affected by ad libitum versus restricted feeding and challenge level. In experiment 1, 144 Cobb 500 males were placed in battery cages with 6 chicks/pen. On d 20, half of the battery pens were placed on feed restriction and all broilers were orally challenged with Eimeria tenella oocysts at one of 3 challenge levels (0, 5,000, or 20,000 sporulated oocysts). Cecal and fecal material were collected separately from d 4 postchallenge through d 10 postchallenge for oocysts output (oocysts shed/g) determination. Six days postchallenge, 3 broilers from each pen were removed and subjected to necropsy for lesion assessment. In experiment 2, 96 Cobb 500 males were placed in identical battery pens with 8 chicks/pen. On d 14, restricted feeding was initiated and broilers were challenged with Eimeria tenella oocysts at one of 3 challenge levels (1,000, 5,000, or 20,000 oocysts). Twenty-four hour collections of cecal and fecal material were obtained separately from d 4 postchallenge through d 10 postchallenge for oocysts per gram and total output determination. Six days postchallenge, 4 broilers from each pen were removed and subjected to necropsy for lesion assessment. In both experiments, BW gain was not affected by challenge dose in either the ad libitum-fed or restrict-fed broilers. Increased lesion development was observed with increasing challenge levels, and oocyst shedding peaked between d 7 and 9 postchallenge in both experiments. Oocyst concentration was higher in cecal droppings compared with fecal material throughout peak shedding; however, total oocyst output for the challenge period was similar between fecal material and cecal droppings.

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

    PubMed

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

    2014-05-01

    Avian coccidiosis is a disease caused by intestinal protozoa in the genus Eimeria. Clinical signs of coccidiosis include intestinal lesions and reduced feed efficiency and BW gain. This 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 examine the differential expression of digestive enzymes, transporters of amino acids, peptides, sugars, and minerals, and an antimicrobial peptide in the small intestine of Eimeria acervulina-infected broilers and layers. Uninfected broilers and layers, in general, expressed these genes at comparable levels. Some differences included 3-fold and 2-fold greater expression of the peptide transporter PepT1 and the antimicrobial peptide LEAP2 (liver expressed antimicrobial peptide 2), respectively, in the jejunum of layers compared with broilers and 17-fold greater expression of LEAP2 in the duodenum of broilers compared with layers. In the duodenum of Eimeria-infected broilers and layers, there was downregulation of aminopeptidase N; sucrase-isomaltase; the neutral, cationic, and anionic amino acid transporters b(o,+)AT/rBAT, B(o)AT, CAT2, and EAAT3; the sugar transporter GLUT2; the zinc transporter ZnT1; and LEAP2. In the jejunum of infected layers there was downregulation of many of the same genes as in the duodenum plus downregulation of PepT1, b(o,+)AT/rBAT, and the y(+) L system amino acid transporters y(+) LAT1 and y(+) LAT2. In the ileum of infected layers there was downregulation of CAT2, y(+)LAT1, the L type amino acid transporter LAT1, and the sugar transporter GLUT1, and upregulation of APN, PepT1, the sodium glucose transporter SGLT4, and LEAP2. In E. acervulina-infected broilers, there were no gene expression changes in the jejunum and ileum. These changes in intestinal digestive enzyme and nutrient transporter gene expression may result in a decrease in the efficiency of protein digestion, uptake of important amino acids

  8. Chemotherapeutic efficacy of sulphadimidine, amprolium, halofuginone and chloroquine phosphate in experimental Eimeria bareillyi coccidiosis of buffaloes.

    PubMed

    Sanyal, P K; Ruprah, N S; Chhabra, M B

    1985-01-01

    Sulphadimidine, amprolium, halofuginone and chloroquine phosphate were administered to buffalo calves 10 days after experimental infection with Eimeria bareillyi. Animals given sulphadimidine or amprolium remained clinically normal and shed only a few oocysts in their faeces. Halofuginone was found partially effective and chloroquine phosphate completely ineffective in preventing faecal oocyst discharge and intestinal lesions. Sulphadimidine and amprolium treated calves gained weight, but chloroquine treated calves suffered progressive weight loss similar to that of infected untreated controls. No significant alterations of haematological values were observed either in the treated calves or in the untreated controls.

  9. Genetic diversity and drug sensitivity studies on Eimeria tenella field isolates from Hubei Province of China.

    PubMed

    Tan, Li; Li, Yalin; Yang, Xin; Ke, Qiyun; Lei, Weiqiang; Mughal, Mudassar Niaz; Fang, Rui; Zhou, Yanqin; Shen, Bang; Zhao, Junlong

    2017-03-09

    Avian coccidiosis is an intracellular intestinal parasitic disease, caused by intracellular intestinal parasites from the genus Eimeria, among which Eimeria tenella is one of the most pathogenic species and causes great economic losses. Frequent applications of anticoccidial drugs have resulted in the development of drug-resistance in E. tenella. In the present study, we sought to determine the genetic diversity of E. tenella isolates prevalent in chicken farms in Hubei Province of China and examine their sensitivity to three anticoccidial drugs. The results provide useful information for the prevention and control of coccidiosis in this region. Eimeria tenella oocysts were isolated from faecal samples collected from different commercial broiler production farms in Hubei Province, China. After oocyst sporulation and animal inoculation for expansion of the field isolates, DNA and RNA were extracted from excysted sporozoites for molecular characterization. Species identification of field isolates were performed by polymerase chain reaction (PCR) amplification of the internal transcribed spacer 1 (ITS1) region of ribosomal DNA. Random amplified polymorphic DNA (RAPD) was used for population genetic analysis. Subsequently, sequences of the major sporozoite surface antigen (SAG), micronemal protein 2 (MIC-2) and cytochrome b (cytb) genes from genomic DNA, and the Eimeria tenella cation-transport ATPase (EtCat ATPase) gene from cDNA were obtained for genotyping using multi-sequence alignments. Finally, sensitivity of the field isolates to three commonly used anticoccidial drugs (diclazuril, decoquinate and maduramycin) were tested to assess the prevalence of drug resistance in E. tenella in Hubei Province of China. Analysis of the ITS1 sequences indicated that all the isolates were E. tenella. RAPD analysis and multi-sequence alignments of the SAG, MIC-2, EtCat ATPase and cytb showed genetic diversity among these isolates. Finally, drug sensitivity tests demonstrated

  10. Resistance to diclazuril in field isolates of Eimeria species obtained from commercial broiler flocks in Brazil.

    PubMed

    Kawazoe, U; Fabio, J D

    1994-06-01

    The efficacy of the anticoccidial drug diclazuril against field isolates of Eimeria spp. collected from broiler farms in four different sites of the South and Southeastern regions of Brazil were investigated. The effect of the drug was measured by an assessment of weight gain, lesion score and oocyst production. Two reference laboratory strains not previously exposed to diclazuril were sensitive to the drug. Two field isolates were judged to be sensitive to diclazuril, four were partly resistant and six were resistant to the drug. The response of the isolates to diclazuril varied depending upon the extent of exposure and type of drug programme in use at the farms.

  11. The effect of high n-3 fatty acids diets on the ultrastructural development of Eimeria tenella.

    PubMed

    Danforth, H D; Allen, P C; Levander, O A

    1997-01-01

    A study of development of Eimeria tenella in chickens fed high n-3 fatty acids (n-3FA) diets showed ultrastructural degeneration of both asexual and sexual parasite stages. Abnormal shedding of asexual and sexual parasite developmental stages into the cecal lumen was also observed. Ultrastructural degeneration was characterized by cytoplasmic vacuolization, chromatin condensation within the nucleus, a lack of parasitophorous vacuole delineation, and, in some cases, a complete loss of parasite ultrastructural organization. The results of this study indicate that diets high in n-3FA may be useful in the control of avian coccidia.

  12. Screening and characterization of apical membrane antigen 1 interacting proteins in Eimeria tenella.

    PubMed

    Han, Hongyu; Xue, Pu; Dong, Hui; Zhu, Shunhai; Zhao, Qiping; Huang, Bing

    2016-11-01

    Avian coccidiosis is a widespread and economically significant disease of poultry. It is an enteric disease caused by several protozoan Eimeria species. Eimeria belongs to the phylum Apicomplexa, which exhibits an unusual mechanism of host cell invasion. During invasion of host cells, the protein apical membrane antigen 1 (AMA1) is essential for invasion of Toxoplasma gondii and Plasmodium. Contrary to the roles of AMA1 during host cell invasion in T. gondii and Plasmodium, the precise functions of Eimeria AMA1 (EtAMA1) are unclear. In order to study the functions of EtAMA1, a yeast two-hybrid cDNA library was constructed from E. tenella sporozoites. The EtAMA1 ectodomain was cloned into the pGBKT7 vector to construct the bait plasmid pGBKT7- EtAMA1. Autoactivation and toxicity of the bait protein in yeast cells were tested by comparison with the pGBKT7 empty vector. Expression of the bait protein was detected by western blots. The bait plasmid pGBKT7-EtAMA1 was used to screen yeast two-hybrid cDNA library from E. tenella sporozoites. After multiple screenings with high-screening-rate medium and exclusion of false-positive plasmids, positive preys were sequenced and analyzed using BLAST. We obtained 14 putative EtAMA1-interacting proteins including E. tenella acidic microneme protein2 (EtMIC2), E. tenella putative cystathionine beta-synthase, E. tenella Eimeria-specific protein, four E. tenella conserved hypothetical proteins (one in the serine/threonine protein kinase family) and seven unknown proteins. Gene Ontology analysis indicated that two known proteins were associated with metabolic process, pyridoxal phosphate binding and protein phosphorylation. Functional analysis indicated EtMIC2 was implicated in parasite motility, migration, recognition and invasion of host cells. The data suggested that EtAMA1 may be important during host cell invasion, but also involved in other biological processes.

  13. [Species from genus Eimeria observed in domestic rabbit (Oryctolagus cuniculus) feces raised at the Municipality of Campos dos Goytacazes in the State of Rio de Janeiro, Brazil].

    PubMed

    de Almeida, Adriana J; Mayen, Friederike L; de Oliveira, Francisco C R

    2006-01-01

    This study was designed to identify species of the genus Eimeria from eleven rabbit meat breeders from Campos dos Goytacazes. Fecal samples were collected from rabbits with different health conditions and consintence of feces. The following species of Eimeria were identified: E. perforans; E. magna; E. coecicola; E. irresidua; E. media; E. flavescens; E. nagpurensis; E. intestinalis, E. exigua and E. stiedae. Parasites from this genus were detected in 81.82% (9) of the rabbit meat production sites, regardless of the management and hygiene conditions. In all cases the infection was always caused from more than one species of Eimeria.

  14. Molecular Characterization and Functional Analysis of a Novel Calcium-Dependent Protein Kinase 4 from Eimeria tenella

    PubMed Central

    Dong, Hui; Zhao, Qiping; Zhu, Shunhai; Xia, Weili; Xu, Shuaibin; Xie, Yuxiang; Cui, Xiaoxia; Tang, Min; Men, Qifei; Yang, Zhiyuang; Li, Cong; Zhu, Xuelong; Han, Hongyu

    2016-01-01

    Eimeria tenella is an obligate intracellular parasite that actively invades cecal epithelial cells of chickens. The basis of cell invasion is not completely understood, but some key molecules of host cell invasion have been discovered. This paper investigated the characteristics of calcium-dependent protein kinase 4 (EtCDPK4), a critical molecule in E. tenella invasion of host cells. A full-length EtCDPK4 cDNA was identified from E. tenella using rapid amplification of cDNA ends. EtCDPK4 had an open reading frame of 1803 bp encoding a protein of 600 amino acids. Quantitative real-time PCR and western blotting were used to explore differences in EtCDPK4 transcription and translation in four developmental stages of E. tenella. EtCDPK4 was expressed at higher levels in sporozoites, but translation was higher in second-generation merozoites. In vitro invasion inhibition assays explored whether EtCDPK4 was involved in invasion of DF-1 cells by E. tenella sporozoites. Polyclonal antibodies against recombinant EtCDPK4 (rEtCDPK4) inhibited parasite invasion, decreasing it by approximately 52%. Indirect immunofluorescence assays explored EtCDPK4 distribution during parasite development after E. tenella sporozoite invasion of DF-1 cells in vitro. The results showed that EtCDPK4 might be important in sporozoite invasion and development. To analyze EtCDPK4 functional domains according to the structural characteristics of EtCDPK4 and study the kinase activity of rEtCDPK4, an in vitro phosphorylation system was established. We verified that rEtCDPK4 was a protein kinase that was completely dependent on Ca2+ for enzyme activity. Specific inhibitors of rEtCDPK4 activity were screened by kinase activity in vitro. Some specific inhibitors were applied to assays of DF-1 cell invasion by E. tenella sporozoites to confirm that the inhibitors functioned in vitro. W-7, H-7, H-89, and myristoylated peptide inhibited DF-1 invasion by E. tenella sporozoites. The experimental results showed

  15. Molecular Characterization and Functional Analysis of a Novel Calcium-Dependent Protein Kinase 4 from Eimeria tenella.

    PubMed

    Wang, Ziwen; Huang, Bing; Dong, Hui; Zhao, Qiping; Zhu, Shunhai; Xia, Weili; Xu, Shuaibin; Xie, Yuxiang; Cui, Xiaoxia; Tang, Min; Men, Qifei; Yang, Zhiyuang; Li, Cong; Zhu, Xuelong; Han, Hongyu

    2016-01-01

    Eimeria tenella is an obligate intracellular parasite that actively invades cecal epithelial cells of chickens. The basis of cell invasion is not completely understood, but some key molecules of host cell invasion have been discovered. This paper investigated the characteristics of calcium-dependent protein kinase 4 (EtCDPK4), a critical molecule in E. tenella invasion of host cells. A full-length EtCDPK4 cDNA was identified from E. tenella using rapid amplification of cDNA ends. EtCDPK4 had an open reading frame of 1803 bp encoding a protein of 600 amino acids. Quantitative real-time PCR and western blotting were used to explore differences in EtCDPK4 transcription and translation in four developmental stages of E. tenella. EtCDPK4 was expressed at higher levels in sporozoites, but translation was higher in second-generation merozoites. In vitro invasion inhibition assays explored whether EtCDPK4 was involved in invasion of DF-1 cells by E. tenella sporozoites. Polyclonal antibodies against recombinant EtCDPK4 (rEtCDPK4) inhibited parasite invasion, decreasing it by approximately 52%. Indirect immunofluorescence assays explored EtCDPK4 distribution during parasite development after E. tenella sporozoite invasion of DF-1 cells in vitro. The results showed that EtCDPK4 might be important in sporozoite invasion and development. To analyze EtCDPK4 functional domains according to the structural characteristics of EtCDPK4 and study the kinase activity of rEtCDPK4, an in vitro phosphorylation system was established. We verified that rEtCDPK4 was a protein kinase that was completely dependent on Ca2+ for enzyme activity. Specific inhibitors of rEtCDPK4 activity were screened by kinase activity in vitro. Some specific inhibitors were applied to assays of DF-1 cell invasion by E. tenella sporozoites to confirm that the inhibitors functioned in vitro. W-7, H-7, H-89, and myristoylated peptide inhibited DF-1 invasion by E. tenella sporozoites. The experimental results showed

  16. Timing relationships among maxima of punch and die-wall stress and punch displacement during compaction of viscoelastic solids.

    PubMed

    Morehead, W T; Rippie, E G

    1990-11-01

    The times of occurrence of maxima in punch stress, die-wall stress, and punch displacement in instrumented rotary tablet machines are shown to be noncoincidental for compacts exhibiting viscoelastic behavior. Equations are presented which characterize materials behaving as Kelvin solids in either or both distortion and dilation. These equations explicitly relate the timing of stress maxima to punch strain and strain rate. Typical effects are illustrated by data obtained for Avicel PH 101, Klucel, and mannitol. Punch stress maxima are shown to significantly precede the time of maximum punch insertion into the die for viscoelastic materials compacted at rates typical of production. Die-wall stress maxima occur after punch stress maxima due to internal rate-limited processes.

  17. Induction of Protective Immunity against Eimeria tenella Infection using Antigen-loaded Dendritic Cells (DC) and DC-derived Exosomes

    USDA-ARS?s Scientific Manuscript database

    Eimeria is a protozoan parasite which causes coccidiosis, a major parasitic disease of poultry that is indigenous to most regions of the world and causes intestinal damage with reduced body weight gain. Conventional disease control methods have relied on prophylactic chemotherapy, but alternative m...

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

    PubMed Central

    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

  19. Eimeria that infect fish are diverse and are related to, but distinct from, those that infect terrestrial vertebrates

    USDA-ARS?s Scientific Manuscript database

    The Eimeria are ubiquitous Apicoplexan parasites (family: coccidia) of the gut epithelium of vertebrates which complete their development in a single host species and whose sporocysts may be recognized by the presence of a Stieda body through which their sporozoites excyst. Their diversity and rel...

  20. Host-Parasite Incongruences in Rodent Eimeria Suggest Significant Role of Adaptation Rather than Cophylogeny in Maintenance of Host Specificity

    PubMed Central

    Kvičerová, Jana; Hypša, Václav

    2013-01-01

    The degree of host specificity, its phylogenetic conservativeness and origin are virtually unknown in Eimeria. This situation is largely due to the inadequate sample of eimerian molecular data available for reliable phylogenetic analyses. In this study, we extend the data set by adding 71 new sequences of coccidia infecting 16 small-mammal genera, mostly rodents. According to the respective feasibility of PCR gene amplification, the new samples are represented by one or more of the following genes: nuclear 18S rRNA, plastid ORF 470, and mitochondrial COI. Phylogenetic analyses of these sequences confirm the previous hypothesis that Eimeria, in its current morphology-based delimitation, is not a monophyletic group. Several samples of coccidia corresponding morphologically to other genera are scattered among the Eimeria lineages. More importantly, the distribution of eimerians from different hosts indicates that the clustering of eimerian species is influenced by their host specificity, but does not arise from a cophylogenetic/cospeciation process; while several clusters are specific to a particular host group, inner topologies within these clusters do not reflect host phylogeny. This observation suggests that the host specificity of Eimeria is caused by adaptive rather than cophylogenetic processes. PMID:23861732

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

    PubMed Central

    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

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

    USDA-ARS?s Scientific Manuscript database

    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...

  3. Comparison of live Eimeria vaccination with in-feed Salinomycin on growth and immune status in broiler chickens

    USDA-ARS?s Scientific Manuscript database

    Coccidiosis vaccines and coccidiostat drugs are commonly used to control Eimeria infection during commercial poultry production. The present study was conducted to compare the relative effectiveness of these two disease control strategies in broiler chickens in an experimental research facility. B...

  4. Induction of Passive Immunity in Broiler Chickens Against Eimeria acervulina by Hyperimmune Egg Yolk IgY

    USDA-ARS?s Scientific Manuscript database

    The protective effect of hyperimmune IgY fraction of egg yolk (SC) prepared from hens hyperimmunized with multiple species of Eimeria oocysts, on experimental coccidiosis was evaluated in young broilers. Chickens were continuously fed from hatch with a standard diet containing SC or a non-suppleme...

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

    PubMed

    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.

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

    PubMed

    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.

  7. Live Eimeria vaccination success in the face of artificial non-uniform vaccine administration in conventionally reared pullets.

    PubMed

    Price, Kayla R; Hafeez, Mian A; Bulfon, Julia; Barta, John R

    2016-01-01

    Live Eimeria vaccines against coccidiosis in poultry initiate immunity using a vaccine dose containing few oocysts; protection is enhanced through subsequent faecal-oral transmission ("cycling") of parasites in the poultry house. Spray-administered Eimeria vaccines can permit wide variations in doses ingested by individual chicks; some chicks may receive no primary vaccination at all. Consequently, protective immunity for the entire flock depends on successful environmental cycling of vaccine progeny. Pullets missing primary vaccination at day of age can become protected from coccidial challenge through cycling of vaccine progeny oocysts from vaccinated (V) cage mates. This study tested whether 40% cage floor coverage (CFC) with a durable material could improve protection against challenge in these "contact-vaccinated" (CV) or successfully V pullets. The six treatment groups tested were CV, V or sham-vaccinated pullets cage-reared on either 0% or 40% CFC. Oocyst output was measured separately for each group for 30 days following vaccine administration. Lesion scores, body weights and total oocyst outputs were measured to quantify protection at 30 days of age against single or mixed Eimeria species challenge infections. Use of 40% CFC to promote low-level oocyst cycling impacted the flock in two ways: (1) more uniform flock immunity was achieved in the 40% CFC (CV similar to V pullets) compared with 0% CFC and (2) protection was enhanced in the 40% CFC compared with the 0% CFC. The use of CFC is an easily adopted means of improving live Eimeria vaccination of caged pullets.

  8. First description of an in vitro culture system for Eimeria ovinoidalis macromeront formation in primary host endothelial cells.

    PubMed

    Carrau, Tessa; Silva, Liliana Machado Ribeiro; Pérez, David; Ruiz de Ybáñez, Rocio; Taubert, Anja; Hermosilla, Carlos

    2016-10-01

    The apicomplexan parasite Eimeria ovinoidalis is distributed worldwide and causes clinical ovine coccidiosis. As one of the most pathogenic species in sheep, the principal clinical sign is profuse diarrhoea in young animals, which leads to important economic losses in the ovine industry. We here aimed to establish an in vitro culture system for the development of E. ovinoidalis macromeronts, as no suitable systems are currently available for any ovine Eimeria species. Faecal samples containing more than 90% of E. ovinoidalis oocysts were collected from naturally infected lambs and ewes in Murcia Region (Spain). E. ovinoidalis oocysts were collected, left to sporulate in potassium dichromate and stored at 4°C until further studies were conducted. Moreover, a suitable excystation protocol was effectively established, resulting in the release of viable sporozoites, which were allowed to infect primary bovine umbilical vein endothelial cells (BUVEC) and permanent bovine colonic epithelial cells (BCEC). In vitro first merogony was successfully accomplished exclusively in BUVEC leading to macromeront formation (up to 100μm) and the release of fully developed and viable merozoites I stages. Given that we were able to establish a suitable in vitro system for the first merogony of such pathogenic Eimeria species in sheep, advances might be further made not only on studies regarding the control of ovine coccidiosis, such as drug screenings, but also on the better understanding of molecular parasite-host cell interactions as already demonstrated for other ruminant Eimeria species.

  9. Pinning down a polymorphic parasite: new genetic and morphological descriptions of Eimeria macropodis from the Tammar wallaby (Macropus eugenii).

    PubMed

    Hill, Nichola J; Richter, Carolin; Power, Michelle L

    2012-09-01

    Identification of the protozoan parasite, Eimeria has traditionally relied on oocyst morphology, host range and life-cycle attributes. However, it is increasingly recognized that Eimeria species can vary in size and shape across their host range, an attribute known as 'polymorphism' that presents a unique challenge for identification. Advances in molecular tools hold promise for characterising Eimeria that may otherwise be misclassified based on morphology. Our study used morphologic and molecular traits of the oocyst life stage to identify a polymorphic parasite, Eimeria macropodis in a captive Tammar wallaby (Macropus eugenii) population in Australia. Molecular characterization highlighted the need to use multiple genetic markers (18S SSU and cytochrome c oxidase subunit I) to accurately identify E. macropodis owing to heterozygous alleles at the 18S SSU locus. This study provided an opportunity to assess the utility and shortcomings of morphologic and molecular techniques for 'pinning down' a polymorphic species. Moreover, our study was able to place E. macropodis in an evolutionary context and enhance resolution of the under-studied marsupial clade.

  10. Is sporozoite refractile body protein expression different in Eimeria acervulina sporozoites isolated from non-immune versus immune chickens?

    USDA-ARS?s Scientific Manuscript database

    A hallmark of Eimeria infection in avians is the establishment of immunity against clinical signs of coccidiosis. Resistant birds experience improved weight gain and feed conversion efficiency and lack intestinal lesions. Oocysts excretion is reduced, but not eliminated, in such immune chickens. ...

  11. Proteomic analysis of Eimeria acervulina sporozoite proteins interaction with duodenal epithelial cells by shotgun LC-MS/MS.

    PubMed

    Zhang, ZhenChao; Wang, Shuai; Huang, JingWei; Liu, LianRui; Lu, MingMin; Li, MengHui; Sui, YuXia; Xu, LiXin; Yan, RuoFeng; Song, XiaoKai; Li, XiangRui

    2015-08-01

    Although it has been known for many years that Eimeria acervulina (E. acervulina) initiates infection by invading the duodenal epithelial cells of chicken, the key protein molecules and the mechanisms of the parasite in invading are unknow. In this study, we found that 85 proteins of E. acervulina could bind with the chicken duodenal epithelial cells from Eimeria protein database. Among them, sixteen were identified only in Eimeria spp. correlation with invasion and evasion and 69 proteins were found in Eimeria spp. with more than 2 unique pep count. Nine out of the 16 proteins and 41 out of the 69 proteins were annotated according to Gene Ontology Annotation in terms of molecular function, biological process, and cellular localization. Most of the 9 annotated proteins occurred in binding, catalytic activity and cellular process whereas, 29 (70.73%) out of the 41 proteins had binding activity and 20 proteins (48.78%) had catalytic activity. The findings provided an insight into the interactive relationship between E. acervulina and host cells and will shed new lights on the understanding of molecular mechanisms of E. acervulina invasion and pathogenesis.

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

    PubMed

    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.

  13. Effects of Artemisia annua and Foeniculum vulgare on on chickens highly infected with Eimeria tenella (Phylum Apicomplexa)

    USDA-ARS?s Scientific Manuscript database

    Background: Intensive poultry production systems depend on chemoprophylaxis with anticoccidial drugs to combat infection. A floor-pen study was conducted to evaluate the anticoccidial effect of Artemisia annua and Foeniculum vulgare on Eimeria tenella infection. Five experimental groups were establi...

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

    PubMed

    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

  15. Physical studies of chromatin. The recombination of histones with DNA.

    PubMed

    Boseley, P G; Bradbury, E M; Butler-Browne, G S; Carpenter, B G; Stephens, R M

    1976-02-02

    Experiments have been carried out to define clearly which histone combinations can induce a higher order structure when combined with DNA. The criterion for a higher order structure being the series of low-angle X-ray diffraction maxima nominally at 5.5 nm, 3.7 nm, 2.7 nm and 2.2 nm. Such a pattern, with resolution similar to that of H1-depleted chromatin, is readily attainable by recombining histones H2A + H2B + H3 + H4 with DNA using a salt-gradient dialysis method. However, the use of urea in the recombination procedure is shown to be detrimental to the production of a higher order structure. Low-angle ring patterns are not obtained by recomgining DNA with single pure histones or any combination of histone pairs exept H3 + H4. The diffraction maxima from the latter are, however, weaker than those from chromatin and there are pronounced semi-equatorial arcs. The presence of a third histone, either H2A or H2B in the H3 + H4 recombination mixture tends to distort the recognised low-angle pattern. It is concluded that the histone pair H3 + H4 is essential for the formation of a regular higher order structure in chromatin, although for a complete structural development the presence of H2A + H2B is also required.

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

    PubMed

    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.

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

    PubMed

    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.

  18. Description of the two strains of turkey coccidia Eimeria adenoeides with remarkable morphological variability.

    PubMed

    Poplstein, Martin; Vrba, Vladimir

    2011-09-01

    Although oocyst morphology was always considered as a reliable parameter for coccidian species discrimination we describe strain variation of turkey coccidia, Eimeria adenoeides, which remarkably exceeds the variation observed in any other Eimeria species. Two strains have been isolated - the first strain maintains the typical oocyst morphology attributed to this species - large and ellipsoidal - while the second strain has small and ovoid oocysts, never described before for this species. Other biological parameters including pathogenicity were found to be similar. Cross-protection between these 2 strains in 2 immunization and challenge experiments was confirmed. Sequencing and analysis of 18S and ITS1 ribosomal DNA revealed a close relationship according to 18S and a relatively distant relationship according to ITS1. Analysis of 18S and ITS1 sequences from commercial turkey coccidiosis vaccines Immucox®-T and Coccivac®-T revealed that each vaccine contains a different strain of E. adenoeides and that these strains have 18S and ITS1 sequences homologous to the sequences of the strains we have isolated and described. These findings show that diagnostics of turkey coccidia according to oocyst morphology have to be carried out with caution or abolished entirely. Novel PCR-based molecular tools will be necessary for fast and reliable species discrimination.

  19. Molecular survey of Apicomplexa in Podarcis wall lizards detects Hepatozoon, Sarcocystis, and Eimeria species.

    PubMed

    Harris, D James; Maia, João P M C; Perera, Ana

    2012-06-01

    The occurrence of apicomplexan parasites in Podarcis sp. wall lizards from the Iberian Peninsula and Balearic islands was studied by amplification and sequencing of the 18S rRNA gene. Species from 3 genera, Hepatozoon , Sarcocystis , and Eimeria , were found. The phylogenetic analysis of the 18S rRNA gene provides unexpected insights into the evolutionary history of these parasites. All Hepatozoon spp. specimens were recovered as part of a clade already identified in lizards from North Africa. The Sarcocystis species, detected in Podarcis lilfordi from Cabrera Island in the Balearic Islands, appears related to Sarcocystis gallotiae , known only from endemic Gallotia sp. lizards from the Canary Islands. Based on the lack of snake predators on this island, this parasite presumably presents an atypical transmission cycle that uses the same host species as both intermediate and final host through cannibalism, like S. gallotiae . Eimeria sp. is reported for the first time from Podarcis spp. lizards. This study shows the power of detecting multiple different apicomplexan parasites through screening of tail tissue samples and blood drops that are often collected in reptiles for other purposes.

  20. Neutrophil extracellular trap formation as innate immune reactions against the apicomplexan parasite Eimeria bovis.

    PubMed

    Behrendt, Jan Hillern; Ruiz, Antonio; Zahner, Horst; Taubert, Anja; Hermosilla, Carlos

    2010-01-15

    Eimeria bovis infections are under immunological control and recent studies have emphasized the role of early PMN-mediated innate immune responses in infected calves. Neutrophil extracellular traps (NETs) have recently been demonstrated to act as a killing mechanism of PMN against several pathogens. In the present study, the interactions of bovine PMN with sporozoites of E. bovis were investigated in this respect in vitro. For demonstration and quantification of NET formation, extracellular DNA was stained by Sytox Orange. Fluorescence images after Sytox Orange staining as well as scanning electron microscopy (SEM) showed NET formation to occur upon contact with E. bovis sporozoites. Exposure of PMN to viable sporozoites induced stronger NET formation than to dead or homogenized parasites. NET formation was abolished by treatment with DNase and could be reduced by diphenylene iodonium, which is described as a potent inhibitor of NADPH oxidase. After sporozoite and PMN co-culture, extracellular fibres were found attached to sporozoites and seemed to trap them, strongly suggesting that NETs immobilize E. bovis sporozoites and thereby prevent them from infecting host cells. Thus, transfer of sporozoites, previously being confronted with PMN, to adequate host cells resulted in clearly reduced infection rates when compared to PMN-free controls. NET formation by PMN may therefore represent an effector mechanism in early innate immune reactions against E. bovis. This is the first report indicating Eimeria-induced NET formation.

  1. Increased level of Eimeria sensitivity to diclazuril after using a live coccidial vaccine.

    PubMed

    Mathis, G F; Broussard, C

    2006-09-01

    Anticoccidial vaccine and an anticoccidial drug rotation program were compared to determine which program was more effective in producing coccidia populations sensitive of 1 ppm diclazuril. The study used an anticoccidial drug-sensitivity battery test (AST) to determine the baseline level of diclazuril sensitivity to field isolates of Eimeria spp. from seven broiler complexes that had used diclazuril. Based on percentage reduction in weight gain and lesion scores, 25% or fewer of the isolates were effectively controlled by diclazuril. Following the baseline sampling, four of the complexes switched to a nondiclazuril in-feed anticoccidial drug program and three of the complexes switched to a vaccination program for two broiler grow-out cycles as the sole coccidiosis-control program. This study demonstrated that the vaccine used (Coccivac-B) contained anticoccidial drug-sensitive strains. Eimeria isolates were subsequently collected from the identical houses and diclazuril AST results were compared with the baseline AST results. Following the two grow-out cycles, sensitivity of the isolates to diclazuril from the four complexes that continued to use in-feed anticoccidial drugs remained essentially unchanged. The isolates from the three complexes that switched to the vaccination program demonstrated a marked increase in diclazuril sensitivity, with 60%-100% of the isolates from each complex effectively controlled by diclazuril. Vaccination with the anticoccidial drug-sensitive strains produced a measurable increase in the level of sensitivity to diclazuril.

  2. Evaluating the Resistance of Eimeria Spp. Field Isolates to Anticoccidial Drugs Using Three Different Indices

    PubMed Central

    Arabkhazaeli, F; Modrisanei, M; Nabian, S; Mansoori, B; Madani, A

    2013-01-01

    Background In this study, the presence of resistance to diclazuril, amprolium+ethopabate and salinomycin, representing some of the commonest anticoccidials in Iran's poultry industry, against three mixed Eimeria field isolates were investigated. Methods Three Eimeria field isolates, collected from typical broiler farms in Iran, were propagated once, inoculated to 480 broilers, comprising 30 chicks in each treatment. The non-medicated or medicated diets containing one of the above mentioned anticoccidials were provided ad-lib. Drug efficacy was determined using the Global index (GI), Anticoccidial Sensitivity Test (AST) and Optimum Anticoccidial Activity (OAA). Results None of the field isolates were fully sensitive to the selected anticoccidials. All isolates showed reduced sensitivity/partial resistance to salinomycin. Resistance to amprolium+ethopabate was evident and partial to complete resistance was recorded for diclazuril. Conclusion Limited efficacy of the selected anticoccidials is obvious. Considering the cost of continuous use of anticoccidials in the field, altering the prevention strategy and rotation of the anticoccidials with better efficacy, would prevent further economic losses induced by coccidiosis. PMID:23914236

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

    PubMed

    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.

  4. Ethanol and isopropanol trigger rapid egress of intracellular Eimeria tenella sporozoites.

    PubMed

    Yan, Xinlei; Liu, Xianyong; Ji, Yongsheng; Tao, Geru; Suo, Xun

    2015-02-01

    Egress from host cells is a vital step of the intracellular life cycle of apicomplexan parasites such as Toxoplasma gondii. This phenomenon has attracted attentions from many research groups. Previous studies have shown that ethanol could stimulate the release of microneme proteins by elevating intracellular Ca(2+) concentration of T. gondii, resulting in the parasite egress from host cells. However, little information about egress is known on Eimeria species, the causative agent of coccidiosis in poultry and livestock. In this report, we studied the effect of ethanol and isopropanol on the egress of eimerian parasites. Eimeria tenella sporozoites cultured in primary chicken kidney cells were treated with ethanol and isopropanol, then the egressed parasites were analyzed. Ethanol and isopropanol could induce the rapid egress of E. tenella sporozoites from host cells. No substantial damage was found in parasite-egressed host cells. Compared to the freshly isolated sporozoites, the re-invading ability and reproductivity of the egressed parasites significantly decreased by 43.4 and 44.1 % individually. We also found that fewer sporozoites egressed from host cells when the parasites developed for a longer time before the alcohol treatment. These results demonstrate an in vitro egress mode different from that of T. gondii, facilitating the deciphering of the mechanisms of egress of eimerian parasites.

  5. Expression of Toxoplasma gondii dense granule protein7 (GRA7) in Eimeria tenella.

    PubMed

    Yin, Guangwen; Qin, Mei; Liu, Xianyong; Suo, Jingxia; Suo, Xun

    2013-05-01

    Dense granules are specialized secretory organelles of Apicomplexa parasites; the dense granule (GRA) proteins are believed to play a role in intracellular survival and the nutrient/waste exchange mechanism with the host cell. Until now, limited information is available concerning the characterization of GRA proteins in Eimeria. Eimeria tenella and Toxoplasma gondii are apicomplexan protozoa and share many similarities in biology and genomics. We hypothesized that GRA proteins from T. gondii could be expressed and have a similar function in E. tenella. To confirm the expression and localization of the GRA protein in T. gondii and E. tenella, a transient transfection strategy was used to express T. gondii GRA7 tagged with yellow fluorescent protein (YFP) (GRA7-YFP); T. gondii tachyzoites were transfected with the plasmid pTgtubGRA7-YFP/sagCAT, and E. tenella sporozoites were transfected with the pEtmic1GRA7-YFP/act construct. The results show that fluorescence can be expressed mainly into the parasitophorous vacuoles (PVs) of the T. gondii. GRA7 of T. gondii can also be expressed in E. tenella and can lead the fluorescence protein into the PVs of the parasites and the cavity of the sporocysts. As for the extracellular stage, YFP gathered to form small particles in the released merozoites and sporozoites, suggesting a localization of the secretory organelles of E. tenella. These results suggest that GRA proteins have a conserved function across species of Apicomplexa in targeting proteins to the PVs.

  6. Anticoccidial effects of the root bark of Dictamnus dasycarpus Turcz extract on experimental Eimeria tenella infection.

    PubMed

    Hong, Sunhwa; Lee, Hyun-A; Kim, Dong-Woo; Oh, Gi-Wook; Kim, Okjin

    2014-12-01

    Anticoccidial effects of the root bark of Dictamnus dasycarpus Turcz (Rutaceae) extract (DDE) were evaluated in chickens following oral infection with Eimeria (E.) tenella. Three-day-old chickens (n=30) were assigned to three groups (control, untreated, and DDE 0.1% treated). Chickens were fed a standard diet supplemented with or without DDE for 1 week prior to infection with E. tenella (10,000 sporulated oocysts per chicken). The effects of DDE on E. tenella infection were assessed by two parameters; fecal oocysts shedding and body weights gain. The DDE-fed chickens produced significantly reduced fecal oocysts (P<0.05) when compared to the E. tenella-infected group fed standard diet. Also, DDE-based diet, improved body weight loss caused by E. tenella infection. Our data demonstrated that DDE had remarkable anticoccidial activities against E. tenella. This finding might have implications for the development of anticoccidial drug. This study is the first to demonstrate anticoccidial effect of DDE on Eimeria parasites.

  7. Anticoccidial effects of the Plantago asiatica extract on experimental Eimeria tenella infection.

    PubMed

    Hong, Sunhwa; Oh, Gi-Wook; Kang, Won-Guk; Kim, Okjin

    2016-03-01

    Anticoccidial effects of the Plantago asiatica extract (PAE) were evaluated in chickens following oral infection with Eimeria (E.) tenella. This study was conducted on the 3-day-old chickens (n=30). Those animals were divided with 3 groups; PAE 0.1% treated/infected (n=10), PAE untreated/infected (n=10) and non-infected control (n=10). Chickens were fed a standard diet supplemented with or without PAE for 1 week prior to infection with E. tenella (10,000 sporulated oocysts per chicken). The effects of PAE on E. tenella infection were assessed by two parameters; fecal oocysts shedding and body weights gain. The PAE-fed chickens produced significantly reduced fecal oocysts (P<0.05) when compared to the E. tenella-infected group fed standard diet. Also, PAE-based diet, improved body weight loss caused by E. tenella infection. Our data demonstrated that PAE had remarkable anticoccidial activities against E. tenella. This finding might have implications for the development of anticoccidial drug. This study is the first to demonstrate anticoccidial effect of PAE on Eimeria parasites.

  8. Stable Transfection of Eimeria intestinalis and Investigation of Its Life Cycle, Reproduction and Immunogenicity

    PubMed Central

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

    2016-01-01

    Rabbit coccidiosis, caused by infection of Eimeria spp. is one of the most severe parasitic diseases in rabbits. Eimeria 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 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. PMID:27303389

  9. Anticoccidial effects of the Plantago asiatica extract on experimental Eimeria tenella infection

    PubMed Central

    Hong, Sunhwa; Oh, Gi-Wook; Kang, Won-Guk

    2016-01-01

    Anticoccidial effects of the Plantago asiatica extract (PAE) were evaluated in chickens following oral infection with Eimeria (E.) tenella. This study was conducted on the 3-day-old chickens (n=30). Those animals were divided with 3 groups; PAE 0.1% treated/infected (n=10), PAE untreated/infected (n=10) and non-infected control (n=10). Chickens were fed a standard diet supplemented with or without PAE for 1 week prior to infection with E. tenella (10,000 sporulated oocysts per chicken). The effects of PAE on E. tenella infection were assessed by two parameters; fecal oocysts shedding and body weights gain. The PAE-fed chickens produced significantly reduced fecal oocysts (P<0.05) when compared to the E. tenella-infected group fed standard diet. Also, PAE-based diet, improved body weight loss caused by E. tenella infection. Our data demonstrated that PAE had remarkable anticoccidial activities against E. tenella. This finding might have implications for the development of anticoccidial drug. This study is the first to demonstrate anticoccidial effect of PAE on Eimeria parasites. PMID:27051444

  10. Biosynthesis and catabolism of mannitol is developmentally regulated in the protozoan parasite Eimeria tenella.

    PubMed

    Allocco, J J; Profous-Juchelka, H; Myers, R W; Nare, B; Schmatz, D M

    1999-04-01

    The mannitol cycle is a metabolic branch of the glycolytic pathway found in Eimeria tenella. In this paper, we describe the biosynthesis and consumption of mannitol during parasite development. Low micromolar levels of mannitol were detected in all of the asexual stages and mannitol production increased sharply during the sexual phase of the life cycle. Unsporulated oocysts had high mannitol content (300 mM or 25% of the oocyst mass). Mannitol-1-phosphate dehydrogenase (M1PDH), the first committed step of the mannitol cycle, was also elevated in sexual stages and this coincides with mannitol levels. Approximately 90% of the mannitol present in unsporulated oocysts was consumed in the first 15 hr of sporulation, and levels continued to drop until the sporulation process was complete at approximately 35 hr. Thus, mannitol appears to be the "fuel" for sporulation during the vegetative stage of the parasite life cycle. Evaluation of oocyst extracts from 6 additional Eimeria species for mannitol content and the presence of M1PDH indicated that the mannitol cycle was broadly present in this genus. This finding combined with the lack of mannitol metabolism in higher eukaryotes makes this pathway an attractive chemotherapeutic target.

  11. Immunomodulatory and antiparasitic effects of garlic extract on Eimeria vermiformis-infected mice.

    PubMed

    Khalil, Atef Mohammed; Yasuda, Masahiro; Farid, Ayman Samir; Desouky, Mohamed Ibrahim; Mohi-Eldin, Mouchira Mohammed; Haridy, Mohie; Horii, Yoichiro

    2015-07-01

    We investigated the immunomodulatory and parasiticidal effects of garlic extract on coccidiosis caused by Eimeria vermiformis infection in male ICR mice. One group received garlic extract daily until the end of the experiment by the oral route from 10 days prior to oral infection with 300 sporulated E. vermiformis oocysts (infected-garlic(+)). The other group served as a control positive with E. vermiformis infection alone (infected-garlic(-)). In the infected-garlic(+) group, garlic extract treatment induced a significant reduction in fecal oocyst output when compared with the infected-garlic(-) group. Histopathological, immunohistochemical, and gene expression analysis for inflammatory cytokines in ileal tissues showed that the garlic extract treatment impaired intracellular development of E. vermiformis during the early stages by increasing the number of intraepithelial CD8(+) T cells and decreasing IL-10 expression. This induced cell cytotoxicity which was reflected by a decrease in oocyst numbers in the intestinal villi and the feces, indicating anticoccidial effects of the garlic extract. However, further studies to explore the precise mechanism of the observed effects of garlic treatment during Eimeria infection are needed to verify our results.

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

    PubMed

    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) ooc