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Molecular evidence on the emergence of benzimidazole resistance SNPs in field isolates of Marshallagia marshalli (Nematoda: Trichostrongylidae) in sheep

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Abstract

The infection with members of the Trichostrongylid nematodes has been frequently reported from sheep and goats. Because of the widespread use of Benzimidazoles (BZs), the resistance suspected to occur in some worms populations. In this study, we focused on the prevalent nematode, Marshallagia marshalli, from the abomasa of sheep. Samples were obtained from at least 10 infected farms and diagnosed with morphological and molecular methods. For resistance analysis, genomic DNA from pooled adult samples of all farms were analysed for the beta tubulin gene to detect any polymorphisms at codon positions of F167Y, E198A and F200Y. According to the results, seven farms (70%) revealed resistance (R) allele at F200Y with relatively high frequency. No other mutations were identified at the other two positions. Also, except for one homozygous (RR) occasion, the isolates with R allele had heterozygous (RS) genotype. This finding indicates that the worm populations are still affected by drugs of the BZ class. However, the genetic data also notes on developing resistance mechanisms in M. marshalli populations in sheep.

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References

  • Altaş MG, Sevgili M, Gökçen A, Aksin N, Bayburs HC (2009) The prevalence of gastro-intestinal nematodes in hair goats of the Sanliurfa region. Turkiye Parazitol Derg 33:20–24

    PubMed  Google Scholar 

  • Alvarez-Sanchez MA, Perez-Garcia J, Cruz-Rojo MA, Rojo-Vazquez FA (2005) Real time PCR for the diagnosis of benzimidazole resistance in trichostrongylids of sheep. Vet Parasitol 129:291–298

    Article  CAS  Google Scholar 

  • Chaudhry U, Miller M, Yazwinski T, Kaplan R, Gilleard J (2014) The presence of benzimidazole resistance mutations in Haemonchus placei from US cattle. Vet Parasitol 204:411–415

    Article  CAS  Google Scholar 

  • Coles GC, Jackson F, Pomroy WE, Prichard RK, von Samson-Himmelstjerna G, Silvestre A, Taylor MA, Vercruysse J (2006) The detection of anthelmintic resistance in nematodes of veterinary importance. Vet Parasitol 136:167–185

    Article  CAS  Google Scholar 

  • Čudeková P, Várady M, Dolinská M, Königová A (2010) Phenotypic and genotypic characterisation of benzimidazole susceptible and resistant isolates of Haemonchus contortus. Vet Parasitol 172:155–159

    Article  Google Scholar 

  • Dallas JF, Irvine R, Halvorsen O, Albon SD (2000) Identification by polymerase chain reaction (PCR) of Marshallagia marshalli and Ostertagia gruehneri from Svalbard reindeer. Int J Parasitol 30:863–866

    Article  CAS  Google Scholar 

  • Elard L, Humbert JF (1999) Importance of the mutation of amino acid 200 of the isotype-1 b-tubulin gene in the benzimidazole-resistance of the small-ruminant parasite, Teladorsagia circumcincta. Parasitol Res 85:452–456

    Article  CAS  Google Scholar 

  • Elard L, Sauve C, Humbert JF (1998) Fitness of benzimidazole-resistant and-susceptible worms of Teladorsagia circumcincta, a nematode parasite of small ruminants. Parasitology 117:571–578

    Article  Google Scholar 

  • Eslami A, Meydani M, Maleki S, Zargarzadeh A (1979) Gastrointestinal nematodes of wild sheep (Ovis orientalis) from Iran. J Wildl Dis 15:263–265

    Article  CAS  Google Scholar 

  • Esteban-Ballesteros M, Rojo-Vázquez FA, Skuce PJ, Melville L, González-Lanza C, Martínez-Valladares M (2017) Quantification of resistant alleles in the β-tubulin gene of field strains of gastrointestinal nematodes and their relation with the faecal egg count reduction test. BMC Vet Res 13:71

    Article  Google Scholar 

  • Falzon LC, Menzies PI, Shakya KP, Jones-Bitton A, Vanleeuwen J, Avula J, Stewart H, Jansen JT, Taylor MA, Learmount J, Peregrine AS (2013) Anthelmintic resistance in sheep flocks in Ontario, Canada. Vet Parasitol 193:150–162

    Article  CAS  Google Scholar 

  • Ghisi M, Kaminsky R, Maser P (2007) Phenotyping and genotyping of Haemonchus contortus isolates reveals a new putative candidate mutation for benzimidazole resistance in nematodes. Vet Parasitol 144:313–320

    Article  CAS  Google Scholar 

  • Heise M, Epe C, Schnieder T (1999) Differences in the second internal transcribed spacer (ITS-2) of eight species of gastrointestinal nematodes of ruminants. J Parasitol 85:431–435

    Article  CAS  Google Scholar 

  • Hoberg EP, Abrams A, Pilitt PA, Jenkins EJ (2012) Discovery and description of a new trichostrongyloid species (Nematoda: Ostertagiinae), abomasal parasites in mountain goat, Oreamnos americanus, from the Western Cordillera of North America. J Parasitol 98:817–846

    Article  Google Scholar 

  • Höglund J, Gustafsson K, Ljungström BL, Engström A, Donnan A, Skuce P (2009) Anthelmintic resistance in Swedish sheep flocks based on a comparison of the results from the faecal egg count reduction test and resistant allele frequencies of the β-tubulin gene. Vet Parasitol 161:60–68

    Article  Google Scholar 

  • Humbert JF, Cabaret J, Elard L, Leignel V, Silvestre A (2001) Molecular approaches to studying benzimidazole resistance in trichostrongylid nematode parasites of small ruminants. Vet Parasitol 101:405–414

    Article  CAS  Google Scholar 

  • Kaplan RM (2004) Drug resistance in nematodes of veterinary importance: a statusreport. Trends Parasitol 20:477–481

    Article  CAS  Google Scholar 

  • Kimura M (1980) A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16:111–120

    Article  CAS  Google Scholar 

  • Kwa MS, Veenstra JG, Roos MH (1994) Benzimidazole resistance in Haemonchus contortus is correlated with a conserved mutation at amino acid 200 in beta-tubulin isotype 1. Mol Biochem Parasitol 63:299–303

    Article  CAS  Google Scholar 

  • Lichtenfels JR, Hoberg EP (1993) The systematics of nematodes that cause ostertagiasis in domestic and wild ruminants in North America: an update and a key to species. Vet Parasitol 46:33–53

    Article  CAS  Google Scholar 

  • Lichtenfels JR, Pilitt PA (1989) Cuticular ridge patterns of Marshallagia marshalli and Ostertagia occidentalis (Nematoda: Trichostrongyloidea) parasitic in ruminants of North America. Proc Helminthol Soc Wash 56:173–182

    Google Scholar 

  • Niciura SCM, Veríssimo CJ, Gromboni JGG, Rocha MIP, de Mello SS, Barbosa CMP, Chiebao DP, Cardoso D, Silva GS, Otsuk IP, Pereira JR (2012) F200Y polymorphism in the β-tubulin gene in field isolates of Haemonchus contortus and risk factors of sheep flock management practices related to anthelmintic resistance. Vet Parasitol 190:608–612

    Article  CAS  Google Scholar 

  • Silvestre A, Cabaret J (2002) Mutation inposition 167 of isotype 1 b-tubulin gene of Trichostrongylid nematodes: role in benzimidazole resistance? Mol Biochem Parasitol 120:297–300

    Article  CAS  Google Scholar 

  • Silvestre A, Humbert JF (2000) A molecular tool for species identification and benzimidazole resistance diagnosis in larval communities of small ruminant parasites. Exp Parasitol 95:271–276

    Article  CAS  Google Scholar 

  • Silvestre A, Cabaret J, Humbert JF (2001) Effect of benzimidazole under-dosing on the resistant allele frequency in Teladorsagia circumcincta (Nematoda). Parasitology 123:103–111

    Article  CAS  Google Scholar 

  • Skuce P, Stenhouse L, Jackson F, Hypša V, Gilleard J (2010) Benzimidazole resistance allele haplotype diversity in United Kingdom isolates of Teladorsagia circumcincta supports a hypothesis of multiple origins of resistance by recurrent mutation. Int J Parasitol 40:1247–1255

    Article  CAS  Google Scholar 

  • Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729

    Article  CAS  Google Scholar 

  • von Samson-Himmelstjerna G, Blackhall WJ, McCarthy JS, Skuce PJ (2007) Single nucleotide polymorphism (SNP) markers for benzimidazole resistance in veterinary nematodes. Parasitology 134:1077–1086

    Article  Google Scholar 

  • von Samson-Himmelstjerna G, Walsh TK, Donnan AA, Carriere S, Jackson F, Skuce PJ, Rohn K, Wolstenholme AJ (2009) Molecular detection of benzimidazole resistance in Haemonchus contortus using real-time PCR and pyrosequencing. Parasitology 136:349–358

    Article  Google Scholar 

  • Waller PJ (1997) Anthelmintic resistance. Vet Parasitol 72:391–412

    Article  CAS  Google Scholar 

  • Walsh TK, Donnan AA, Jackson F, Skuce P, Wolstenholme AJ (2007) Detection and measurement of benzimidazole resistance alleles in Haemonchus contortus using real-time PCR with locked nucleic acid Taqman probes. Vet Parasitol 144:304–312

    Article  CAS  Google Scholar 

  • Winterrowd CA, Pomroy WE, Sangster NC, Johnson SS, Geary TG (2003) Benzimidazole-resistant beta-tubulin alleles in a population of parasitic nematodes (Cooperia oncophora) of cattle. Vet Parasitol 117:161–172

    Article  CAS  Google Scholar 

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Acknowledgements

The authors would thank Mr. Amir Mootabi Alavi for his kind assistance in collection of samples.

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This study was supported by a Grant from by Shiraz University, Iran (No. FF97).

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Correspondence to Ehsan Rakhshandehroo.

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Fakhrahmad, F., Rakhshandehroo, E. & Ghaemi, M. Molecular evidence on the emergence of benzimidazole resistance SNPs in field isolates of Marshallagia marshalli (Nematoda: Trichostrongylidae) in sheep. J Parasit Dis 45, 435–440 (2021). https://doi.org/10.1007/s12639-020-01319-0

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