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Morphological and molecular characterization of three Eimeria species from captured rangeland goats in Western Australia

机译：西澳大利亚州捕获的牧场山羊中三种艾美球虫物种的形态和分子特征

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Faecal shedding of Eimeria by captured rangeland goats (Capra hircus) was investigated using a longitudinal observational study. Faecal samples were collected from 125 male goats on four occasions. The first sampling occurred following capture and transport, immediately after arrival at a commercial goat depot (feedlot) in Western Australia, with subsequent 3 sample collections occurring at one month intervals thereafter. Goats were composite breed and aged approximately 9–12 months on arrival at the feedlot. Prevalence and shedding intensity (faecal oocyst concentration) for Eimeria were determined using qPCR. Species were identified from individual oocysts (isolated using micromanipulation) using molecular analysis at two loci, specifically 18S rRNA and mitochondrial cytochrome oxidase gene (COI), and confirmed by microscopy. Longitudinal prevalence (animals positive at least once) for Eimeria spp. by qPCR was 90.4%, with 60% goats shedding Eimeria spp. on more than one occasion. Point prevalence (prevalence at a single sampling occasion) ranged from 2.4% (fourth sampling) to 70.4% (second sampling). Three species were identified at the 18S rRNA locus and confirmed by microscopy: E. christenseni (longitudinal prevalence for single infection 34.4%), E. hirci (17.6%) and E. arloingi (8.8%) over the four sample collections. Mixed infections were identified in 56.8% goats (longitudinal prevalence). 18S rRNA sequences from E. christenseni and E. hirci were 100% homologous with ovine E. ahsata and E. crandallis respectively, and E. arloingi was 100% similar to caprine E. arloingi. At the COI locus, E. christenseni, E. hirci and E. arloingi grouped separately, and were closely related to ovine E. ahsata, with genetic similarities of 96.5%, 92.6% and 91.4% respectively. This is the first report for molecular characteristics of caprine-derived Eimeria spp. using a combination of 18S rRNA and COI. Molecular techniques can be used to identify Eimeria spp. in goat faecal samples, specifically through characterization at 18S locus and other gene loci when used in parallel. Molecular techniques offer some advantages over microscopy for identification of Eimeria species, particularly with respect to precision.
机译：使用纵向观察研究调查了捕获的牧场山羊（Capra hircus）对艾美球虫的粪便脱落。四次从125只公山羊中收集粪便样品。第一次采样是在捕获和运输之后进行的，紧接着到达澳大利亚西部的一个商业山羊仓库（饲养场）后，随后以一个月为间隔进行了3次采样。山羊是复合品种，到达饲养场时大约9-12个月大。使用qPCR确定艾美球虫的发生率和脱落强度（粪便卵囊浓度）。使用两个位点，特别是18S rRNA和线粒体细胞色素氧化酶基因（COI）的分子分析，从单个卵囊（通过显微操作分离）中鉴定出物种，并通过显微镜确认。 Eimeria spp的纵向流行率（动物至少一次阳性）。通过qPCR的结果是90.4％，其中60％的山羊脱落了艾美球虫属。不止一次。点患病率（单次采样发生率）范围从2.4％（第四次采样）到70.4％（第二次采样）。在18S rRNA基因座上鉴定出3个物种，并通过显微镜确认：克里斯滕森大肠杆菌（单次感染的纵向患病率为34.4％），埃希氏大肠杆菌（17.6％）和埃罗希氏大肠杆菌（8.8％）在四个样本集合中。在56.8％的山羊中发现混合感染（纵向流行）。来自christenseni和E. hirci的18S rRNA序列分别与羊的E. ahsata和E. crandallis同源性为100％，而E. arloingi与Caprine E. arloingi相似性为100％。在COI位点，E。christenseni，E。hirci和E. arloingi分别分组，并且与绵羊的ahsata紧密相关，遗传相似性分别为96.5％，92.6％和91.4％。这是关于山羊衍生的艾美球虫属的分子特性的首次报道。使用18S rRNA和COI的组合。分子技术可用于鉴定艾美球虫。特别是通过在18S基因座和其他基因座上同时鉴定时，对山羊粪便样品进行鉴定。分子技术相对于显微镜在鉴定艾美尔球虫物种方面具有一些优势，特别是在精度方面。

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