Phenotypic and genotypic analysis of benzimidazole resistance in reciprocal genetic crosses of Haemonchus contortus.
Phenotypic and genotypic analysis of benzimidazole resistance in reciprocal genetic crosses of Haemonchus contortus.
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DOI：
10.1016/j.ijpddr.2021.11.001
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发表时间：
2022-04
期刊：
影响因子：
--
通讯作者：
Bartley DJ
中科院分区：
文献类型：
--
作者：
Morrison AA;Chaudhry U;Andrews L;Melville L;Doyle SR;Sargison ND;Bartley DJ
捻转血矛线虫可以说是全球范围内经济上最重要和普遍存在的家畜寄生虫之一，并且通常涉及驱虫剂抗性的情况。在这里，我们使用敏感（MHco 3（ISE））和多重驱虫药抗性（MHco 18（UGA 2004））H进行了相互遗传杂交。扭体线虫分离株所得混合种群被命名为MHco 3/18或MHco 18/3，其中先导分离株反映了雌性的来源。每个杂交组合产生3个独立的子代，通过生物测定、分子生物学方法和群体遗传学分析，研究了各时期苯并咪唑（BZ）抗性的表型和基因型遗传。一组微卫星标记证实了遗传杂交的成功，因为在F1杂交中看到了来自双亲的标记。卵孵化试验表明，两个F1杂交种之间存在明显差异，MHco18/3的ED 50估计值是MHco3/18的9倍。基于ED 50估计的抗性因子在子代中为MHco3（ISE）亲本的6至57倍。利用焦磷酸测序和MiSeq深度扩增子测序对与BZ抗性相关的F167 Y和F200 Y SNP标记进行分子分析，结果显示MHco3/18.F1和MHco18/3.F1具有相似的F200 Y抗性等位基因频率（分别为45.3%和44.3%），而对于F167 Y，MHco18/3.F1的抗性等位基因频率是MHco3/18.F1的两倍（分别为18.2%和8.8%）。焦磷酸测序和MiSeq扩增子测序之间的比较显示，来自两种方法的等位基因频率在密码子200处是一致的（rc = 0.97），但是对于密码子167不太可比（rc = 0.55）。利用受控的遗传互作揭示了BZ抗性表型的潜在差异取决于抗性等位基因是父系遗传还是母系遗传。这些发现提供了新的见解，并促进进一步调查的遗传BZ抗性在H。扭曲用于研究苯并咪唑（BZ）耐药性的交叉试验。结合表型和基因型工具进行分析。BZ抗性遗传受母体和/或细胞质机制影响。在β-微管蛋白基因上检测到F167 Y和F200 Y的双纯合抗性基因型。
Haemonchus contortus is arguably one of the most economically important and ubiquitous parasites of livestock globally and commonly involved in cases of anthelmintic resistance. Here, we performed reciprocal genetic crosses using susceptible (MHco3(ISE)) and multiple anthelmintic resistant (MHco18(UGA2004)) H. contortus isolates. Resultant admixed populations were designated MHco3/18 or MHco18/3, where the lead isolate reflects the origin of the females. Three independent filial generations were generated for each cross, which were subjected to bioassays, molecular approaches and population genetic analyses to investigate the phenotypic and genotypic inheritance of benzimidazole (BZ) resistance at each stage. A panel of microsatellite markers confirmed the success of the genetic cross as markers from both parents were seen in the F1 crosses. Egg hatch tests revealed a stark difference between the two F1 crosses with ED50 estimates for MHco18/3 being 9 times greater than those for MHco3/18. Resistance factors based on ED50 estimates ranged from 6 to 57 fold in the filial progeny compared to MHco3(ISE) parents. Molecular analysis of the F167Y and F200Y SNP markers associated with BZ resistance were analysed by pyrosequencing and MiSeq deep amplicon sequencing, which showed that MHco3/18.F1 and MHco18/3.F1 both had similar frequencies of the F200Y resistant allele (45.3% and 44.3%, respectively), whereas for F167Y, MHco18/3.F1 had a two-fold greater frequency of the resistant-allele compared to MHco3/18.F1 (18.2% and 8.8%, respectively). Comparison between pyrosequencing and MiSeq amplicon sequencing revealed that the allele frequencies derived from both methods were concordant at codon 200 (rc = 0.97), but were less comparable for codon 167 (rc = 0.55). The use of controlled reciprocal genetic crosses have revealed a potential difference in BZ resistance phenotype dependent on whether the resistant allele is paternally or maternally inherited. These findings provide new insight and prompt further investigation into the inheritance of BZ resistance in H. contortus. Reciprocal cross used to investigate benzimidazole (BZ) resistance. Phenotypic and genotypic tools combined for analysis. Inheritance of BZ resistance influenced by maternal &/or cytoplasmic mechanisms. Double homozygous resistant genotypes at F167Y and F200Y detected on β−tubulin gene.
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DOI：
10.1016/j.ijpddr.2014.07.007
发表时间：
2014-12
影响因子：
4
作者：
Kotze, Andrew C.;Hunt, Peter W.;Skuce, Philip;von Samson-Himmelstjerna, Georg;Martin, Richard J.;Sager, Heinz;Kruecken, Juergen;Hodgkinson, Jane;Lespine, Anne;Jex, Aaron R.;Gilleard, John S.;Beech, Robin N.;Wolstenholme, Adrian J.;Demeler, Janina;Robertson, Alan P.;Charvet, Claude L.;Neveu, Cedric;Kaminsky, Ronald;Rufener, Lucien;Alberich, Melanie;Menez, Cecile;Prichard, Roger K.
通讯作者：
Prichard, Roger K.
影响因子：
2
作者：
Kerboeuf, D;Chambrier, P;Aycardi, J
通讯作者：
Aycardi, J
DOI：
10.1016/j.ijpddr.2012.02.003
发表时间：
2012-12-01
影响因子：
4
作者：
Kotze, Andrew C.;Cowling, Katie;Coleman, Glen T.
通讯作者：
Coleman, Glen T.
DOI：
10.1093/bioinformatics/bts199
发表时间：
2012-06-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Kearse M;Moir R;Wilson A;Stones-Havas S;Cheung M;Sturrock S;Buxton S;Cooper A;Markowitz S;Duran C;Thierer T;Ashton B;Meintjes P;Drummond A
通讯作者：
Drummond A
影响因子：
2.6
作者：
Excoffier, Laurent;Laval, Guillaume;Schneider, Stefan
通讯作者：
Schneider, Stefan