{"title":"窄头棘虫β-微管蛋白同型的特征及其对钩虫苯并咪唑抗性的影响","authors":"Thomas Stocker , Jan Šlapeta","doi":"10.1016/j.vetpar.2025.110569","DOIUrl":null,"url":null,"abstract":"<div><div><em>Uncinaria stenocephala</em> is a widespread hookworm of dogs across Europe, Canada, southern Australia, and other temperate regions, where it often outnumbers infections caused by <em>Ancylostoma caninum</em>. Although a putative β-tubulin isotype-1 mutation associated with resistance has been detected in <em>U. stenocephala</em>, clinical resistance to benzimidazoles has not yet been confirmed. Benzimidazole resistance is primarily linked to single nucleotide polymorphisms (SNPs) in the β-tubulin isotype-1 gene; however, the β-tubulin genes of <em>U. stenocephala</em> have not been fully characterised. We aimed to identify β-tubulin genes and confirm the coding sequences for key residues (Q134, F167, E198, and F200) in the β-tubulin isotype-1 gene of the <em>U. stenocephala</em> genome. Two <em>U. stenocephala</em> specimens were subjected to Illumina sequencing, and species identity was confirmed through morphological and molecular analysis using ITS rDNA and <em>cox</em>1 markers. Genome assembly revealed the presence of β-tubulin isotype-1 (10 exons) and isotype-2 (9 exons), both homologous to β-tubulins from other hookworms (<em>A. caninum</em>, <em>A. ceylanicum</em>, <em>A. duodenale</em> and <em>Necator americanus</em>). The β-tubulin isotype-1 protein sequence of <em>U. stenocephala</em> contained two variable residues (S37Q and G441A) compared to other hookworm sequences. While the isotype-2 protein sequence was conserved among <em>Ancylostoma</em> species, <em>U. stenocephala</em> exhibited six distinct polymorphisms (E39D, T40S, N115S, L130I, A287S, T439G). The benzimidazole-susceptible residues (Q134, F167, E198, F200) were present in the β-tubulin isotype-1 protein sequence. Characterisation of the complete coding regions of β-tubulin isotypes 1 and 2 enables population-level screening for benzimidazole resistance-associated SNPs and provides a foundation for future epidemiological studies in <em>U. stenocephala</em>.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"339 ","pages":"Article 110569"},"PeriodicalIF":2.2000,"publicationDate":"2025-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Characterisation of β-tubulin isotypes in Uncinaria stenocephala and implications for benzimidazole resistance in hookworms\",\"authors\":\"Thomas Stocker , Jan Šlapeta\",\"doi\":\"10.1016/j.vetpar.2025.110569\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div><em>Uncinaria stenocephala</em> is a widespread hookworm of dogs across Europe, Canada, southern Australia, and other temperate regions, where it often outnumbers infections caused by <em>Ancylostoma caninum</em>. Although a putative β-tubulin isotype-1 mutation associated with resistance has been detected in <em>U. stenocephala</em>, clinical resistance to benzimidazoles has not yet been confirmed. Benzimidazole resistance is primarily linked to single nucleotide polymorphisms (SNPs) in the β-tubulin isotype-1 gene; however, the β-tubulin genes of <em>U. stenocephala</em> have not been fully characterised. We aimed to identify β-tubulin genes and confirm the coding sequences for key residues (Q134, F167, E198, and F200) in the β-tubulin isotype-1 gene of the <em>U. stenocephala</em> genome. Two <em>U. stenocephala</em> specimens were subjected to Illumina sequencing, and species identity was confirmed through morphological and molecular analysis using ITS rDNA and <em>cox</em>1 markers. Genome assembly revealed the presence of β-tubulin isotype-1 (10 exons) and isotype-2 (9 exons), both homologous to β-tubulins from other hookworms (<em>A. caninum</em>, <em>A. ceylanicum</em>, <em>A. duodenale</em> and <em>Necator americanus</em>). The β-tubulin isotype-1 protein sequence of <em>U. stenocephala</em> contained two variable residues (S37Q and G441A) compared to other hookworm sequences. While the isotype-2 protein sequence was conserved among <em>Ancylostoma</em> species, <em>U. stenocephala</em> exhibited six distinct polymorphisms (E39D, T40S, N115S, L130I, A287S, T439G). The benzimidazole-susceptible residues (Q134, F167, E198, F200) were present in the β-tubulin isotype-1 protein sequence. Characterisation of the complete coding regions of β-tubulin isotypes 1 and 2 enables population-level screening for benzimidazole resistance-associated SNPs and provides a foundation for future epidemiological studies in <em>U. stenocephala</em>.</div></div>\",\"PeriodicalId\":23716,\"journal\":{\"name\":\"Veterinary parasitology\",\"volume\":\"339 \",\"pages\":\"Article 110569\"},\"PeriodicalIF\":2.2000,\"publicationDate\":\"2025-08-05\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Veterinary parasitology\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0304401725001803\",\"RegionNum\":2,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"PARASITOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Veterinary parasitology","FirstCategoryId":"97","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0304401725001803","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"PARASITOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
窄头钩虫是一种广泛存在于欧洲、加拿大、南澳大利亚和其他温带地区的狗钩虫,在这些地区,它的感染人数经常超过由犬钩虫引起的感染人数。虽然在窄头乌中发现了与耐药相关的β-微管蛋白同型1突变,但临床对苯并咪唑的耐药尚未得到证实。苯并咪唑耐药性主要与β-微管蛋白同型1基因的单核苷酸多态性(snp)有关;然而,小头鲸的β-微管蛋白基因尚未被完全表征。我们的目的是鉴定小头鲸β-微管蛋白基因,并确定β-微管蛋白同型1基因中关键残基(Q134、F167、E198和F200)的编码序列。利用ITS rDNA和cox1标记进行形态和分子分析,确认了两份窄头乌标本的物种身份。基因组组装显示存在β-微管蛋白同型1(10个外显子)和同型2(9个外显子),它们与其他钩虫(A. caninum, A. ceylanicum, A. duodenale和Necator americanus)的β-微管蛋白同源。与其他钩虫序列相比,窄头棘球绦虫β-微管蛋白同型1序列含有两个可变残基(S37Q和G441A)。虽然同型2蛋白序列在各物种间具有保守性,但窄头棘球绦虫具有6种不同的多态性(E39D、T40S、N115S、L130I、A287S、T439G)。β-微管蛋白同型1序列中存在苯并咪唑敏感残基Q134、F167、E198、F200。β-微管蛋白同型1和2的完整编码区特征可以在人群水平上筛选与苯并咪唑耐药相关的snp,并为未来的窄头棘球蚴流行病学研究提供基础。
Characterisation of β-tubulin isotypes in Uncinaria stenocephala and implications for benzimidazole resistance in hookworms
Uncinaria stenocephala is a widespread hookworm of dogs across Europe, Canada, southern Australia, and other temperate regions, where it often outnumbers infections caused by Ancylostoma caninum. Although a putative β-tubulin isotype-1 mutation associated with resistance has been detected in U. stenocephala, clinical resistance to benzimidazoles has not yet been confirmed. Benzimidazole resistance is primarily linked to single nucleotide polymorphisms (SNPs) in the β-tubulin isotype-1 gene; however, the β-tubulin genes of U. stenocephala have not been fully characterised. We aimed to identify β-tubulin genes and confirm the coding sequences for key residues (Q134, F167, E198, and F200) in the β-tubulin isotype-1 gene of the U. stenocephala genome. Two U. stenocephala specimens were subjected to Illumina sequencing, and species identity was confirmed through morphological and molecular analysis using ITS rDNA and cox1 markers. Genome assembly revealed the presence of β-tubulin isotype-1 (10 exons) and isotype-2 (9 exons), both homologous to β-tubulins from other hookworms (A. caninum, A. ceylanicum, A. duodenale and Necator americanus). The β-tubulin isotype-1 protein sequence of U. stenocephala contained two variable residues (S37Q and G441A) compared to other hookworm sequences. While the isotype-2 protein sequence was conserved among Ancylostoma species, U. stenocephala exhibited six distinct polymorphisms (E39D, T40S, N115S, L130I, A287S, T439G). The benzimidazole-susceptible residues (Q134, F167, E198, F200) were present in the β-tubulin isotype-1 protein sequence. Characterisation of the complete coding regions of β-tubulin isotypes 1 and 2 enables population-level screening for benzimidazole resistance-associated SNPs and provides a foundation for future epidemiological studies in U. stenocephala.
期刊介绍:
The journal Veterinary Parasitology has an open access mirror journal,Veterinary Parasitology: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review.
This journal is concerned with those aspects of helminthology, protozoology and entomology which are of interest to animal health investigators, veterinary practitioners and others with a special interest in parasitology. Papers of the highest quality dealing with all aspects of disease prevention, pathology, treatment, epidemiology, and control of parasites in all domesticated animals, fall within the scope of the journal. Papers of geographically limited (local) interest which are not of interest to an international audience will not be accepted. Authors who submit papers based on local data will need to indicate why their paper is relevant to a broader readership.
Parasitological studies on laboratory animals fall within the scope of the journal only if they provide a reasonably close model of a disease of domestic animals. Additionally the journal will consider papers relating to wildlife species where they may act as disease reservoirs to domestic animals, or as a zoonotic reservoir. Case studies considered to be unique or of specific interest to the journal, will also be considered on occasions at the Editors'' discretion. Papers dealing exclusively with the taxonomy of parasites do not fall within the scope of the journal.