PDF(Pubmed)
Abstract：
BACKGROUND: Wild deer populations utilizing livestock grazing areas risk cross-species transmission of gastrointestinal nematode parasites (GINs), including GINs with anthelmintic resistance (AR) traits. Wild deer have been shown to carry problematic GIN species such as Haemonchus contortus and Trichostrongylus species in the UK, but the presence of livestock GINs in Northern Ireland deer populations is unknown. Also, is it not known whether AR traits exist among GINs of deer such as Ostertagia leptospicularis and Spiculopteragia asymmetrica in pastureland where anthelmintics are heavily used.
METHODS: Adult-stage GIN samples were retrieved from Northern Irish wild fallow deer abomasa. Individual specimens were subject to a species-specific PCR analysis for common sheep and cattle GIN species with ITS-2 sequence analysis to validate species identities. In addition, the beta-tubulin gene was subject to sequencing to identify benzimidazole (BZ) resistance markers.
RESULTS: ITS-2 sequencing revealed O. leptospicularis and S. asymmetrica, but species-specific PCR yielded false-positive hits for H. contortus, Teladorsagia circimcincta, Trichostrongylus axei, T. colubriformis, T. vitrinus and Ostertagia ostertagi. For beta-tubulin, O. leptospicularis and S. asymmetrica yielded species-specific sequences at the E198 codon, but no resistance markers were identified in either species at positions 167, 198 or 200 of the coding region.
CONCLUSIONS: From this report, no GIN species of significance in livestock were identified among Northern Ireland fallow deer. However, false-positive PCR hits for sheep and cattle-associated GINs is concerning as the presence of deer species in livestock areas could impact both deer and livestock diagnostics and lead to overestimation of both GIN burden in deer and the role as of deer as drivers of these pathogens. ITS-2 sequences from both O. leptospicularis and S. asymmetrica show minor sequence variations to geographically distinct isolates. AR has been noted among GINs of deer but molecular analyses are lacking for GINs of wildlife. In producing the first beta-tubulin sequences for both O. leptospicularis and S. asymmetrica, we report no BZ resistance in this cohort.
CONCLUSIONS: This work contributes to genetic resources for wildlife species and considers the implications of such species when performing livestock GIN diagnostics.
METHODS: Adult-stage GIN samples were retrieved from Northern Irish wild fallow deer abomasa. Individual specimens were subject to a species-specific PCR analysis for common sheep and cattle GIN species with ITS-2 sequence analysis to validate species identities. In addition, the beta-tubulin gene was subject to sequencing to identify benzimidazole (BZ) resistance markers.
RESULTS: ITS-2 sequencing revealed O. leptospicularis and S. asymmetrica, but species-specific PCR yielded false-positive hits for H. contortus, Teladorsagia circimcincta, Trichostrongylus axei, T. colubriformis, T. vitrinus and Ostertagia ostertagi. For beta-tubulin, O. leptospicularis and S. asymmetrica yielded species-specific sequences at the E198 codon, but no resistance markers were identified in either species at positions 167, 198 or 200 of the coding region.
CONCLUSIONS: From this report, no GIN species of significance in livestock were identified among Northern Ireland fallow deer. However, false-positive PCR hits for sheep and cattle-associated GINs is concerning as the presence of deer species in livestock areas could impact both deer and livestock diagnostics and lead to overestimation of both GIN burden in deer and the role as of deer as drivers of these pathogens. ITS-2 sequences from both O. leptospicularis and S. asymmetrica show minor sequence variations to geographically distinct isolates. AR has been noted among GINs of deer but molecular analyses are lacking for GINs of wildlife. In producing the first beta-tubulin sequences for both O. leptospicularis and S. asymmetrica, we report no BZ resistance in this cohort.
CONCLUSIONS: This work contributes to genetic resources for wildlife species and considers the implications of such species when performing livestock GIN diagnostics.
摘要：
背景：利用牲畜放牧区的野鹿种群有可能跨物种传播胃肠道线虫寄生虫（GINs），包括具有驱虫抗性（AR）特征的GINs。在英国，野鹿已被证明携带有问题的GIN物种，例如扭曲的Haemonchus和Trichostrongylus物种，但是北爱尔兰鹿群中是否存在牲畜gins是未知的。此外，尚不清楚在大量使用驱虫药的牧场中，鹿的母鹿中是否存在AR特征，例如Ostertagialeptopspicularis和Spiculopterifrica不对称。
方法：成人阶段的GIN样本是从北爱尔兰野生休耕鹿寄养的。对单个标本进行物种特异性PCR分析，对常见的绵羊和牛GIN物种进行ITS-2序列分析，以验证物种身份。此外,对β-微管蛋白基因进行测序以鉴定苯并咪唑(BZ)抗性标记。
结果：ITS-2测序显示O.钩皮球和S.不对称，但是物种特异性PCR对H.contortus产生了假阳性，Teladorsagiacirimcincta，阿西毛线菌,T.Colubriformis,T.玻璃化和Ostertagiaostertagi。对于β-微管蛋白，O.leptospularis和S.asymmetrica在E198密码子处产生了物种特异性序列，但是在编码区的167、198或200位的任何一个物种中均未发现抗性标记。
结论：从本报告中，在北爱尔兰的小鹿中，没有发现在牲畜中具有重要意义的GIN物种。然而,与绵羊和牛相关的GINs的假阳性PCR命中令人担忧，因为牲畜区域中鹿物种的存在可能会影响鹿和牲畜的诊断，并导致对鹿的GIN负担以及鹿作为这些病原体的驱动因素的作用的高估。来自O.钩皮囊和S.不对称的ITS-2序列显示出与地理上不同的分离株的微小序列变异。在鹿的GIN中已经注意到AR，但缺乏对野生动物GIN的分子分析。在产生O.leptospicularis和S.asymmetrica的第一个β-微管蛋白序列时，我们报告该队列中没有BZ耐药.
结论：这项工作有助于野生动物物种的遗传资源，并考虑了这些物种在进行家畜GIN诊断时的影响。
方法：成人阶段的GIN样本是从北爱尔兰野生休耕鹿寄养的。对单个标本进行物种特异性PCR分析，对常见的绵羊和牛GIN物种进行ITS-2序列分析，以验证物种身份。此外,对β-微管蛋白基因进行测序以鉴定苯并咪唑(BZ)抗性标记。
结果：ITS-2测序显示O.钩皮球和S.不对称，但是物种特异性PCR对H.contortus产生了假阳性，Teladorsagiacirimcincta，阿西毛线菌,T.Colubriformis,T.玻璃化和Ostertagiaostertagi。对于β-微管蛋白，O.leptospularis和S.asymmetrica在E198密码子处产生了物种特异性序列，但是在编码区的167、198或200位的任何一个物种中均未发现抗性标记。
结论：从本报告中，在北爱尔兰的小鹿中，没有发现在牲畜中具有重要意义的GIN物种。然而,与绵羊和牛相关的GINs的假阳性PCR命中令人担忧，因为牲畜区域中鹿物种的存在可能会影响鹿和牲畜的诊断，并导致对鹿的GIN负担以及鹿作为这些病原体的驱动因素的作用的高估。来自O.钩皮囊和S.不对称的ITS-2序列显示出与地理上不同的分离株的微小序列变异。在鹿的GIN中已经注意到AR，但缺乏对野生动物GIN的分子分析。在产生O.leptospicularis和S.asymmetrica的第一个β-微管蛋白序列时，我们报告该队列中没有BZ耐药.
结论：这项工作有助于野生动物物种的遗传资源，并考虑了这些物种在进行家畜GIN诊断时的影响。
