Trichostrongyles are gastrointestinal parasites that occur globally and can cause subclinical to severe, sometimes life-threatening, infections in ruminants, particularly young animals. Benzimidazoles (BZ) are commonly used for the treatment of gastrointestinal parasites in ruminants. Increasing spread of worm populations with anthelmintics resistance has been reported and is considered a consequence of highly frequent and longstanding use of anthelmintics. To obtain initial information regarding the occurrence of putatively BZ-resistant Nigerian Haemonchus populations, screening based on the molecular analysis of BZ-resistance-associated β-tubulin isotype 1 gene sequence polymorphisms was undertaken. Genomic DNA was isolated from pooled adult Haemonchus sp. from 35 animals from each of the six states of southwestern Nigeria. Sequencing of internal transcribed spacer 2 (ITS-2) and external transcribed spacer (ETS) regions was used to determine the Haemonchus species. Pyrosequencing assays were used for detection of single-nucleotide polymorphisms (SNPs) in the β-tubulin isotype 1 genes of the worms at codons 200 and 167 (TTC/TAC) or 198 (GAA/GCA). Exclusively, Haemonchus placei was detected and allele frequencies obtained at all three positions showed no evidence for the presence of resistance-related alleles. For Lagos State, pools of 10 worms from 30 different animals were analyzed separately for the codon 200 SNP, successfully excluding the presence of resistance-associated SNPs in very low frequencies. These positive findings, showing absence of elevated frequencies of BZ-resistance-associated β-tubulin alleles, have considerable significance since it suggests that farmers can still rely on the efficacy of this important drug class when used for controlling trichostrongyle infections in cattle in Nigeria.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Ademola IO (2002) A survey on Ivermectin resistance in strongyles of sheep in Oyo State, Nigeria, using a larval development assay. Isr J Vet Med 57:149–151
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410
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
Barrere V, Keller K, von Samson-Himmelstjerna G, Prichard RK (2013) Efficiency of a genetic test to detect benzimidazole resistant Haemonchus contortus nematodes in sheep farms in Quebec, Canada. Parasitol Int 62:464–470
Chaudhry U, Miller M, Yazwinski T, Kaplan R, Gilleard J (2014a) The presence of benzimidazole resistance mutations in Haemonchus placei from US cattle. Vet Parasitol 204:411–415
Chaudhry U, Redman EM, Abbas M, Raman M, Ashraf K, Gilleard JS (2014b) Genetic evidence for hybridization between Haemonchus contortus and Haemonchus placei in natural field populations and its implications for interspecies transmission of anthelmintic resistance. Int J Parasitol. doi:10.1016/j.ijpara.2014.09.002
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
Demeler J, Van Zeveren AM, Kleinschmidt N, Vercruysse J, Hoglund J, Koopmann R, Cabaret J, Claerebout E, Areskog M, von Samson-Himmelstjerna G (2009) Monitoring the efficacy of ivermectin and albendazole against gastrointestinal nematodes of cattle in Northern Europe. Vet Parasitol 160:109–115
Demeler J, Ramünke S, Wolken S, Ianiello D, Rinaldi L, Gahutu JB, Cringoli G, von Samson-Himmelstjerna G, Krücken J (2013) Discrimination of gastrointestinal nematode eggs from crude fecal egg preparations by inhibitor-resistant conventional and real-time PCR. PLoS One 8:e61285
Elard L, Humbert JF (1999) Importance of the mutation of amino acid 200 of the isotype 1 beta-tubulin gene in the benzimidazole resistance of the small-ruminant parasite Teladorsagia circumcincta. Parasitol Res 85:452–456
Encalada-Mena L, Tuyub-Solis H, Ramirez-Vargas G, Mendoza-de-Gives P, Aguilar-Marcelino L, Lopez-Arellano ME (2014) Phenotypic and genotypic characterisation of Haemonchus spp. and other gastrointestinal nematodes resistant to benzimidazole in infected calves from the tropical regions of Campeche State, Mexico. Vet Parasitol 205:246–254
Gasbarre LC, Smith LL, Lichtenfels JR, Pilitt PA (2009) The identification of cattle nematode parasites resistant to multiple classes of anthelmintics in a commercial cattle population in the US. Vet Parasitol 166:281–285
Gasser RB, Zhu X, Chilton NB, Newton LA, Nedergaard T, Guldberg P (1998) Analysis of sequence homogenisation in rDNA arrays of Haemonchus contortus by denaturing gradient gel electrophoresis. Electrophoresis 19:2391–2395
Ghisi M, Kaminsky R, Mäser 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
Gibbs HC, Herd RP (1986) Nematodiasis in cattle. Importance, species involved, immunity, and resistance. Veterinary Clin North Am Food Anim Pract 2:211–224
Kenyon F, Greer AW, Coles GC, Cringoli G, Papadopoulos E, Cabaret J, Berrag B, Varady M, Van Wyk JA, Thomas E, Vercruysse J, Jackson F (2009) The role of targeted selective treatments in the development of refugia-based approaches to the control of gastrointestinal nematodes of small ruminants. Vet Parasitol 164:3–11
Mbah AH, Ogunrinade AF, Dina OA (1992) Benzimidazole resistance in strongyles of sheep in Ibadan, Nigeria. Afr Livest Res 11:16–17
Mejia ME, Fernandez Igartua BM, Schmidt EE, Cabaret J (2003) Multispecies and multiple anthelmintic resistance on cattle nematodes in a farm in Argentina: the beginning of high resistance? Vet Res 34:461–467
Prichard R (2001) Genetic variability following selection of Haemonchus contortus with anthelmintics. Trends Parasitol 17:445–453
Prichard RK, von Samson-Himmelstjerna G, Blackhall WJ, Geary TG (2007) Foreword: towards markers for anthelmintic resistance in helminths of importance in animal and human health. Parasitology 134:1073–1076
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
Silvestre A, Leignel V, Berrag B, Gasnier N, Humbert JF, Chartiere C, Cabaret J (2002) Sheep and goat nematode resistance to anthelmintics: pro and cons among breeding management factors. Vet Res 33:465–480
Tiwari J, Kumar S, Kolte AP, Swarnkar CP, Singh D, Pathak KM (2006) Detection of benzimidazole resistance in Haemonchus contortus using RFLP-PCR technique. Vet Parasitol 138:301–307
Tiwari J, Kolte AP, Kumar S, Swarnkar CP, Singh D, Pathak KML (2007) Comparative efficacy of AS-PCR and RFLP-PCR techniques for detection of benzimidazole resistance in Haemonchus contortus of sheep. J Vet Parasitol 21:101–104
van Wyk JA (2001) Refugia–overlooked as perhaps the most potent factor concerning the development of anthelmintic resistance. Onderstepoort J Vet Res 68:55–67
von Samson-Himmelstjerna G (2006) Molecular diagnosis of anthelmintic resistance. Vet Parasitol 136:99–107
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
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
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
Wolstenholme AJ, Fairweather I, Prichard R, von Samson-Himmelstjerna G, Sangster NC (2004) Drug resistance in veterinary helminths. Trends Parasitol 20:469–476
Zarlenga DS, Stringfellow F, Nobary M, Lichtenfels JR (1994) Cloning and characterization of ribosomal RNA genes from three species of Haemonchus (Nematoda: Trichostrongyloidea) and identification of PCR primers for rapid differentiation. Exp Parasitol 78:28–36
The first author is grateful to The World Academy of Sciences for the Advancement of Science in Developing Countries (TWAS) for providing travel cost and local expenses and the German Research Foundation (DFG) for funding his sustenance in Germany. The University of Ibadan, Nigeria, is thanked for granting the main author research leave. The experiments comply with the current laws in Germany.
About this article
Cite this article
Ademola, I.O., Krücken, J., Ramünke, S. et al. Absence of detectable benzimidazole-resistance associated alleles in Haemonchus placei in cattle in Nigeria revealed by pyrosequencing of β-tubulin isotype 1. Parasitol Res 114, 1997–2001 (2015). https://doi.org/10.1007/s00436-015-4406-2
- Haemonchus placei
- Anthelmintic resistance
- Single-nucleotide polymorphism (SNP)