Use of a reverse line blot assay to survey small strongyle (Strongylida: Cyathostominae) populations in horses before and after treatment with ivermectin
Use of a reverse line blot assay to survey small strongyle (Strongylida: Cyathostominae) populations in horses before and after treatment with ivermectin
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DOI：
10.1016/j.vetpar.2009.11.021
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发表时间：
2010-03-25
影响因子：
2.6
通讯作者：
Yeargan, Michelle
中科院分区：
文献类型：
--
作者：
Ionita, Mariana;Howe, Daniel K.;Yeargan, Michelle
采用灵敏、特异的聚合酶链式反应杂交技术，对驱虫药伊维菌素治疗前后的马群中的小口圆线虫种群进行了种属特异性监测。在治疗前和治疗后每周从8匹马(4匹小马驹和4岁的幼马)身上收集6周的粪便样本，以确定每克粪便中的斯特拉伊尔卵数(EPGs)。此外，一只小马驹和一岁的小马驹是未经处理的对照组。此外，一匹来自另一个已知感染了Strongylus spp.的马群的马是这些寄生虫的阳性对照。基因组DNA从大约6000-7000个鸡蛋组中获得，除了两个含有低EPG的样品，其中使用450个鸡蛋。通过扩增马的核糖体DNA(RDNA)基因间隔区(IGSs)，然后进行反向线条杂交(RIB)分析，鉴定了12个最常见的马属小种，并将它们与Strongylus spp.进行了区分。总体而言，在前处理样品中鉴定出11个小型斯特拉尔物种。在伊维菌素治疗后4周采集的样本中，共鉴定出8种小型强虫，其中4种为优势种(长囊环虫、长囊环虫、杯状环虫和微小环刺虫)。在治疗后5周和6周，RLB分析显示，小壮族人群中的成分与治疗前几乎相同。Strongylus spp.仅在从阳性对照马采集的样本中鉴定出这些寄生虫。这些数据证实了聚合酶链式反应-限制性片段长度多态性技术能够同时对马斯特里尔虫卵进行物种特异性分化，为进一步研究抗药性提供了一条有价值的途径。(C)2009年爱思唯尔BM。版权所有。
A sensitive and specific PCR hybridization assay was applied for species-specific monitoring of the small strongyle (Strongylida: Cyathostominae) populations in horses in a herd before and after treatment with the anthelmintic drug ivermectin. Fecal samples were collected pre- and post-treatment weekly from eight individual horses (four foals and four yearlings) for 6 weeks to determine counts of strongyle eggs per gram of feces (EPGs). Additionally, one foal and one yearling were nontreated controls. Also, one horse, from another herd known to be infected with Strongylus spp., was a positive control for these parasites. Genomic DNA was obtained from eggs in groups of approximately 6000-7000 eggs except for two samples containing low EPGs in which 450 eggs were used. Amplification of the intergenic spacers (IGSs) of ribosomal DNA (rDNA) of small and large strongyles followed by reverse line blot (RIB) assay were performed to identify the presence of the 12 most common equine small strongyle species and to discriminate them from Strongylus spp. Overall, 11 small strongyle species were identified in pretreatment samples. In the samples collected at 4 weeks after ivermectin treatment, eight small strongyle species were identified and four of them were predominant (Cylicocyclus nassatus, Cylicostephanus longibursatus, Cylicostephanus calicatus and Cylicostephanus minutus). At 5 and 6 weeks post-treatment, the RLB assay analysis showed almost the same composition in the small strongyle population as before treatment. Strongylus spp. were identified only in samples collected from the positive control horse for these parasites. These data confirm the ability of the PCR-RLB technique for simultaneous species-specific differentiation of equine strongyle eggs, indicating a valuable way of furthering drug-resistance studies. (C) 2009 Elsevier BM. All rights reserved.