Department of Veterinary Biosciences, Melbourne Veterinary School, Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Werribee, Victoria, Australia.
Parasit Vectors. 2018 Jun 28;11(1):370. doi: 10.1186/s13071-018-2963-9.
Gastrointestinal nematodes (GINs) frequently infect South American camelids (alpacas and llamas) and cause economic losses due to reduced production of fiber, meat and/or leather. Our knowledge about the epidemiology and diagnosis of GINs in llamas and alpacas is limited, and reliable keys for the identification of the third-stage larvae (L3s) of some common nematodes (such as Camelostrogylus mentulatus) that infect alpacas and llamas remain undescribed. In this study, we modified two existing semi-quantitative multiplexed-tandem (MT)-PCR assays, originally developed for the GINs of sheep and cattle, to reliably detect and differentiate the common genera/species of GINs in the faeces of alpacas.
Following the establishment of the MT-PCR assay using positive and negative control samples, alpaca faecal samples were tested to validate the assay to detect and differentiate nematode genera/species, including C. mentulatus, Cooperia spp., Haemonchus spp., Oesophagostomum spp., Ostertagia ostertagi, Teladorsagia circumcincta and Trichostrongylus spp. Sequencing of the MT-PCR products demonstrated specific (100%) amplification of the target nematode genera/species. Additionally, a comparison of results of the MT-PCR assay and the morphological identification of adult worms collected from the same 35 alpacas revealed that there was a good agreement (37-94%) between the two methods. However, some discrepancies were observed between the results of the MT-PCR assay and the morphological identification of adult worms.
The MT-PCR platform is an accurate, sensitive and rapid method for the diagnosis of GINs in alpacas, and it can be used as a substitute to larval culture to identify common nematodes in the faeces of alpacas and llamas.
胃肠道线虫（GINs）经常感染南美骆驼（羊驼和骆马），并因纤维、肉和/或皮革产量减少而造成经济损失。我们对骆马和羊驼 GINs 的流行病学和诊断知之甚少，并且对于鉴定感染羊驼和骆马的一些常见线虫（如 Camelostrogylus mentulatus）的第三期幼虫（L3）的可靠钥匙仍然未被描述。在这项研究中，我们修改了两个现有的半定量多重串联（MT）-PCR 检测方法，最初是为绵羊和牛的 GINs 开发的，以可靠地检测和区分羊驼粪便中的常见 GINs 属/种。
在使用阳性和阴性对照样品建立 MT-PCR 检测方法后，测试了羊驼粪便样品以验证该检测方法可检测和区分线虫属/种，包括 C. mentulatus、Cooperia spp.、Haemonchus spp.、Oesophagostomum spp.、Ostertagia ostertagi、Teladorsagia circumcincta 和 Trichostrongylus spp.。MT-PCR 产物的测序显示了目标线虫属/种的特异性（100%）扩增。此外，将 MT-PCR 检测方法的结果与从同一 35 只羊驼收集的成虫形态鉴定结果进行比较表明，两种方法之间存在良好的一致性（37-94%）。然而，在 MT-PCR 检测方法的结果与成虫形态鉴定之间观察到一些差异。
MT-PCR 平台是一种准确、敏感和快速的方法，可用于诊断羊驼的 GINs，它可以替代幼虫培养，用于鉴定羊驼和骆马粪便中的常见线虫。
