In Australia, Cooperia spp. are often overshadowed by parasites believed to be more pathogenic production-limiting nematodes. A rise in anthelmintic resistance and reports of reduced growth rates attributed to infection with Cooperia spp. in Europe increases the need to be able to monitor the presence of C. pectinata, C. punctata and C. oncophora in Australian cattle. Here, we present the first molecular confirmation of C. pectinata and C. punctata in Australian cattle using ITS2 rDNA and COXII mtDNA. Cultured larvae were morphologically differentiated to the genus level with the aid of iodine solution and their DNA was screened using a cattle nematode MT-PCR panel. By isolating individual iodine stained and morphologically identified nematode larvae, we demonstrated the presence of C. pectinata and C. punctata using a generic ITS2 rDNA qPCR assay following DNA amplicon sequencing. A novel suite of COXII mtDNA species/genus-specific PCR assays for Cooperia speciation from complex nematode samples enabled us to detect all three species (C. oncophora, C. pectinata, C. punctata) in Australia cattle samples. Our approach, utilising traditional techniques coupled with the manipulation of individual nematode larvae, provides a foundation for the inclusion of Cooperia spp. into existing high throughput molecular diagnostic panels for cattle nematode surveillance.

中文翻译：

---

他们是哪个幼虫？使用单个幼虫对澳大利亚牛的Cooperia pectinata和Cooperia punctata进行分子鉴定。

在澳大利亚，Cooperia spp。通常被认为是更多致病性生产限制线虫的寄生虫所掩盖。驱虫抗药性增加，以及因库珀菌属感染引起的增长率降低的报道。在欧洲，人们越来越需要能够监测澳大利亚牛中的果胶衣梭菌，点状梭菌和oncophora。在这里，我们介绍了使用ITS2 rDNA和COXII mtDNA在澳大利亚牛中的C. pectinata和C. punctata的首次分子确证。借助碘溶液将培养的幼虫在形态上分化到属水平，并使用牛线虫MT-PCR面板筛选其DNA。通过分离单个碘染色并在形态上鉴定的线虫幼虫，我们证明了果胶梭菌和果蝇的存在。DNA扩增子测序后，使用通用的ITS2 rDNA qPCR测定法检测点子。针对复杂线虫样品中库珀物种形成的一套新颖的COXII mtDNA物种/属特异性PCR分析试剂盒，使我们能够检测澳大利亚牛样品中的所有三种物种（C. oncophora，C。pectinata，C。punctata）。我们的方法，利用传统技术结合对单个线虫幼虫的操纵，为库珀菌种的加入奠定了基础。进入现有的用于牛线虫监视的高通量分子诊断面板。利用传统技术结合对单个线虫幼虫的操纵，为库珀木瓜的混入奠定了基础。进入现有的用于牛线虫监视的高通量分子诊断面板。利用传统技术结合对单个线虫幼虫的操纵，为库珀木瓜的混入奠定了基础。进入现有的用于牛线虫监视的高通量分子诊断面板。