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ABSTRACT: Abstract : The prevalence of coccidial infections in dairy cattle was examined in Shanghai from November 2010 to March 2011. In total, 626 fecal samples from 24 dairy farms were examined; oocysts were identified to the species level based on morphological features. All herds were infected with Eimeria species. The overall prevalence of coccidia was 47.1%, with the highest prevalence in <4-mo-old calves (51.8%) and the lowest in >12-mo-old cattle (27.0%). The number of oocysts per gram of feces was significantly higher in young calves than in weaners and adults. Ten species of Eimeria were identified, among which Eimeria ellipsoidalis , Eimeria bovis , Eimeria zuernii , and Eimeria alabamensis were the predominant species. Concurrent infection with 2-8 species was common.
Journal of Parasitology 05/2012; 98(5):963-6. · 1.40 Impact Factor
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Hui Dong,
Chunhua Li, Qiping Zhao,
Jing Li,
Hongyu Han,
Lianlian Jiang,
Shunhai Zhu,
Ting Li,
Chunlin Kong,
Bing Huang,
Jinzhong Cai
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ABSTRACT: Abstract : 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.
Journal of Parasitology 04/2012; 98(5):958-62. · 1.40 Impact Factor
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ABSTRACT: Apical membrane antigen-1 (AMA1) is a micronemal protein of apicomplexan parasites that appears to be essential during the invasion of host cells. In this study, a full-length cDNA of AMA1 was identified from Eimeria tenella (Et) using expressed sequence tag and the rapid amplification of cDNA ends technique. EtAMA1 had an open reading frame of 1608 bp encoding a protein of 535 amino acids. Quantitative real-time PCR analysis revealed that EtAMA1 was expressed at higher levels in sporozoites than in the other developmental stages (unsporulated oocysts, sporulated oocysts and second-generation merozoites). The ectodomain sequence was expressed as recombinant EtAMA1 (rEtAMA1) and rabbit polyclonal antibodies raised against the rEtAMA1 recognized a 58-kDa native parasite protein by Western Blotting and had a potent inhibitory effect on parasite invasion, decreasing it by approximately 70%. Immunofluorescence analysis and immunohistochemistry analysis showed EtAMA1 might play an important role in sporozoite invasion and development.
PLoS ONE 01/2012; 7(7):e41115. · 4.09 Impact Factor
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ABSTRACT: Serine protease inhibitors (serpins) mediate many biological processes, including immune responses to pathogenic infection. In this study, a member of the serpin superfamily was identified from the common poultry parasite Eimeria tenella by expressed sequence tag analysis and the rapid amplification of cDNA ends technique. The full-length cDNA was 1,918 bp and had an open reading frame of 1,248 bp encoding a polypeptide of 415 amino acids with the theoretical isoelectric point of 5.26 and predicted molecular weight of 45.5 kDa. Real-time quantitative PCR analysis revealed that the serpin gene was expressed at higher levels in sporozoites than in the other developmental stages (unsporulated oocysts, sporulated oocysts, and second-generation merozoites). The sequence encoding the mature protein was amplified by PCR, cloned into the pET28(a) vector, and expressed in Escherichia coli. Specific antiserum generated against the recombinant protein was prepared and used to determine invasion inhibition capacity and localization; the results suggested that the serpin may play an important role in invasion and survival of the sporoziotes in the host.
Parasitology Research 08/2011; 110(2):865-74. · 2.15 Impact Factor
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ABSTRACT: 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.
Parasitology Research 11/2010; 108(4):1033-40. · 2.15 Impact Factor
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ABSTRACT: In order to study the functions of the HSPs (Heat shock proteins) of Eimeria tenella, we cloned a novel gene (which designated EtHSP) coding HSP of Eimeria tenella by RT-PCR and RACE (Rapid-amplification of cDNA ends). The full-length cDNA sequence of EtHSP was 1802 bp, containing a 1455 bp ORF (Open reading frame) (GenBank Accession No. FJ911605) encoding a deduced protein of 484 amino acids. Real-time PCR revealed that the mRNA level of EtHSP was much higher in sporozoites of E. tenella than other developmental stages (unsporulated oocysts, sporulated oocysts and merozoites). We constructed the recombinant plasmids pET28a(+)-EtHSP, then transformed it into E. coli BL21(DE3) for expression. SDS-PAGE indicated that the fusion protein was expressed in included bodies, with peak expression 6 h after induction by IPTG Western blotting revealed that the protein was specifically recognized by polyclonal antibodies against E. tenella, showing that the fusion protein was native antigen.
Sheng wu gong cheng xue bao = Chinese journal of biotechnology 08/2009; 25(8):1121-9.
