Skip to main content
Log in

Molecular detection of benzimidazole resistance levels associated with F167Y and F200Y polymorphisms in Haemonchus contortus of goats from Mozambique

Parasitology Research Aims and scope Submit manuscript

Abstract

Benzimidazole (BZ) resistance of Haemonchus contortus has been associated with single nucleotide polymorphisms (SNPs) in codons 200 (F200Y) and 167 (F167Y) and, to a lesser extent, in codon E198A, of the β-tubulin isotype 1 gene. The present study was undertaken to survey the status of BZ resistance in naturally infected goats in smallholder farms in southern Mozambique by real-time PCR (qPCR) using TaqMan® assays. H. contortus-infective larvae (L3; n = 432) from 12 populations were individually genotyped for F200Y and F167Y SNPs to detect BZ resistance. For the F200Y SNP, the results revealed an overall mean percentages of 18.8% homozygous resistant (RR), 47.8% homozygous susceptible (SS) and 33.4% heterozygous (RS) H. contortus. For the F167Y SNP, the overall mean percentages were 1.6% RR, 94.9% SS and 3.5% RS. The percentage of resistant alleles (%R) for the F200Y and F167Y SNPs was 35.7 and 3.4%, respectively. Genotype combinations of the two mutations indicate resistant percentages ranging from 0.0 to 52.9%. From the four herds with high RR individuals, three farms dewormed the animals monthly, while the fourth farm dewormed the animals every 3 months. In farms where animals were dewormed every 6 months, low percentages of RR individuals were found, whereas no RR individuals were discovered in herds where animals were dewormed annually. These results suggest that the F200Y SNP is more significant in BZ resistance development of the surveyed population compared with the F167Y SNP.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

References

  • Adediran O A, Uwalaka, E C (2015) Effectiveness evaluation of levamisole, albendazole, ivermectin, and Vernonia amygdalina in West African dwarf goats. J Parasitol Res 2015, Article ID 706824, 5 pp.

    Article  CAS  Google Scholar 

  • Akkari H, Jebali J, Gharbi M, Mhadhbi M, Awadi S, Darghouth MA (2013) Epidemiological study of sympatric Haemonchus species and genetic characterization of Haemonchus contortus in domestic ruminants in Tunisia. Vet Parasitol 193:118–125

    Article  CAS  PubMed  Google Scholar 

  • Atanásio A, Boomker J, Sitoe C (2002) A survey on the occurrence of resistance to anthelmintics of gastrointestinal nematodes of goats in Mozambique. Onderstepoort J Vet Res 69:215–220

    PubMed  Google Scholar 

  • Atanásio-Nhacumbe A, Ayres MCC, Lambert SM, Souza BP (2017) Anthelmintic resistance in gastrointestinal nematodes of goats in southern Mozambique. J Vet Med Anim Health 9:313–319

    Google Scholar 

  • Barrère V, Alvarez L, Suarez G, Ceballos L, Moreno L, Lanusse C, Prichard RK (2012) Relationship between increased albendazole systemic exposure and changes in single nucleotide polymorphisms on the β-tubulin isotype 1 encoding gene in Haemonchus contortus. Vet Parasitol 186:344–349

    Article  CAS  PubMed  Google Scholar 

  • Barrère V, Falzon LC, Shakya KP, Menzies PI, Peregrine AS, Prichard RK (2013) Assessment of benzimidazole resistance in Haemonchus contortus in sheep flocks in Ontario, Canada: comparison of detection methods for drug resistance. Vet Parasitol 198:159–165

    Article  CAS  PubMed  Google Scholar 

  • Chagas ACS, Katiki LM, Silva IC, Giglioti R, Esteves SN, Oliveira MCS, Barrioni Junior W (2013) Haemonchus contortus: a multiple-resistant Brazilian isolate and the costs for its characterization and maintenance for research use. Parasitol Int 62:1–6

    Article  PubMed  Google Scholar 

  • Chandra S, Prasad A, Yadav N, Latchumikanthan A, Rakesh RI, Praveen K, khobra V, Subramani KV, Misri J, Sankar M (2015) Status of benzimidazole resistance in Haemonchus contortus of goats from different geographic regions of Uttar Pradesh, India. Vet. Parasitol 208:263–267

    Article  CAS  PubMed  Google Scholar 

  • Chaudhry U, Redman EM, Raman M, Gilleard JS (2015) Genetic evidence for the spread of a benzimidazole resistance mutation across southern India from a single origin in the parasitic nematode Haemonchus contortus. Int J Parasitol 45:721–728

    Article  CAS  PubMed  Google Scholar 

  • Chaudhry U, Redman EM, Ashraf K, Shabbir MZ, Rashid MI, Ashraf S, Gilleard JS (2016) Microsatellite marker analysis of Haemonchus contortus populations from Pakistan suggests that frequent benzimidazole drug treatment does not result in a reduction of overall genetic diversity. Parasit Vectors 9:349–360

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Coles GC, Bauer C, Borgsteede FHM, Geerts TR, Klei MA, Taylor A, Waller PJ (1992) World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) methods for the detection of anthelmintic resistance in nematodes of veterinary importance. Vet Parasitol 44:35–44

    Article  CAS  PubMed  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  PubMed  Google Scholar 

  • Diniz MA, Bandeira S, Eurico S, Martins ES (2012) Flora and vegetation of Maputo province: its appropriation by populations. In: Proceedings of international congress on tropical knowledge in Mozambique: history, memory and science, IICT-JBT, pp 1–2

    Google Scholar 

  • Echevarria FAM, Armour J, Dunca JL (1991) Efficacy of some anthelmintics on an ivermectin resistant strain of Haemonchus contortus in sheep. Vet Parasitol 39:279–284

    Article  CAS  PubMed  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 13:71–78

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gelcer E, Fraisse CW, Zotarelli L, Stevens FR, Perondi D, Barreto DD, Malia HA, Ecole CC, Montone V, Southwort J (2017) Influence of El Nino-Southern Oscillation (ENSO) on agroclimatic zoning for tomato in Mozambique. Agric For Meteorol 248:316–328

    Article  Google Scholar 

  • Ghisi M, Kaminshy 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

    Article  CAS  PubMed  Google Scholar 

  • Gilleard JS (2013) Haemonchus contortus as a paradigm and model to study anthelmintic drug resistance. Parasitology 140:1506–1522

    Article  CAS  PubMed  Google Scholar 

  • Gilleard JS, Beech RS (2007) Population genetics of anthelmintic resistance in parasitic nematodes. Parasitology 134:1133–1147

    Article  CAS  PubMed  Google Scholar 

  • Gupta SK, Singla LD (2012) Diagnostic trends in parasitic diseases of animals. In: Gupta RP, Garg SR, Nehra V, Lather D (eds) Veterinary diagnostics: current trends. Satish Serial Publishing House, Delhi, pp 81–112

    Google Scholar 

  • Kotze AC, Cowling K, Bagnall NH, Hine B, Ruffell AP, Hunt PW, Coleman GT (2012) Relative level of thiabendazole resistance associated with the E198A and F200Y SNPs in larvae of a multi-drug resistant isolate of Haemonchus contortus. Int J Parasitol Drugs Drug Resist. 2:92–97

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lambert SM, Nishi SM, Mendonça LR, Souza BMP, Julião FS, Gusmão PS, Almeida MAO (2017) Genotypic profile of benzimidazole resistance associated with SNP F167Y and F200Y in beta-tubulin gene in Brazilian populations of Haemonchus contortus of goats. Vet Parasitol Reg Stud Rep 8:28–34

    Google Scholar 

  • Leathwick DM, Besier RB (2014) The management of anthelmintic resistance in grazing ruminants in Australasia—strategies and experiences. Vet Parasitol 204:44–54

    Article  CAS  PubMed  Google Scholar 

  • McArthur MJ, Reinemeyer CR (2014) Herding the U.S. cattle industry towards a paradigm shift in parasite control. Vet Parasitol 204:34–43

    Article  CAS  PubMed  Google Scholar 

  • McKellar QA, Jackson F (2004) Veterinary anthelmintics: old and new. Trends Parasitol 20:456–461

    Article  CAS  PubMed  Google Scholar 

  • Melo ACFL, Bevilaqua CML (2005) Genetic approach of anthelmintic resistance in Haemonchus contortus: a review article. Revista Portuguesa de Ciências Veterinárias 100:141–146

    Google Scholar 

  • Mohanraj K, Subhadra S, Kalyanasundaram A, Ilangopathy M, Raman M (2016) Genotyping of benzimidazole resistant and susceptible isolates of Haemonchus contortus from sheep by allele specific PCR. J Parasitic Dis. 7pp. Online: http://www.researchgate.net/publication/304184063

  • Nunes RL, Dos Santos LL, Bastianetto E, Oliveira DAA, Brasil BSAF (2013) Frequency of benzimidazole resistance in Haemonchus contortus populations isolated from buffalo, goat and sheep herds. Rev Bras Parasitol Vet 22:548–553

    Article  CAS  PubMed  Google Scholar 

  • Papadopoulos E (2008) Anthelmintic resistance in sheep nematodes. Small Ruminant Res 76:99–103

    Article  Google Scholar 

  • Peña-Espinoza M, Thamsborg SM, Demeler J, Enemar KH (2014) Field efficacy of four anthelmintics and confirmation of drug-resistant nematodes by controlled efficacy test and pyrosequencing on a sheep and goat farm in Denmark. Vet Parasitol 206:208–215

    Article  CAS  PubMed  Google Scholar 

  • Ramünke S, Melville L, Rinaldi L, Hertzberg H, Waal T, von Samson-Himmelstjerna G, Cringoli G, Mavrot F, Skuce F, Krücken J, Demeler J (2016) Benzimidazole resistance survey of Haemonchus, Teladorsagia and Trichostrongylus in three European countries using pyrosequencing including the development of new assays for Trichostrongylus. Int J Parasitol Drugs Drug Resist. 6:230–240

    Article  PubMed  PubMed Central  Google Scholar 

  • Rüfener L, Kaminsky R, Mäser P (2009) In vitro selection of Haemonchus contortus for benzimidazole resistance reveals a mutation at amino acid 198 of β-tubulin. Mol Biochem Parasitol 168:120–122

    Article  CAS  PubMed  Google Scholar 

  • Salgado JA, Santos CP (2016) Overview of anthelmintic resistance of gastrointestinal nematodes of small ruminants in Brazil. Rev Brasileira Parasitol Vet 25

  • Shija DSN, Kusiluka LJM, Chenyambuga SW, Shayo D, Lekule FP (2014) Animal health constraints in dairy goats kept under smallholder farming systems in Kongwa and Mvomero Districts, Tanzania. J Vet Med Anim Health 6:268–279

    Article  Google Scholar 

  • Singh E, Kaur P, Singla LD, Bal MS (2017a) Prevalence of gastrointestinal parasitism in small ruminants in western zone of Punjab, India. Vet World 10:61–66

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Singh R, Bal MS, Singla LD, Kaur P (2017b) Detection of anthelmintic resistance in sheep and goats against fenbendazole by faecal egg count reduction test. J Parasit Dis 41:463–466

    Article  PubMed  Google Scholar 

  • Tiwari J, Kumar S, Kolte AP, Swarnkar CP, Singh D, Pathak KML (2006) Detection of benzimidazole resistance in Haemonchus contortus using RFLP-PCR technique. Vet Parasitol 138:301–307

    Article  CAS  PubMed  Google Scholar 

  • Ueno H, Gonçalves PC (1998) Handbook for diagnosis of helminthoses of ruminants, 4th edn. Japan International Cooperation, Tokyo, p 143

    Google Scholar 

  • Von Samson-Himmelstjerna G, Walsh TK, Donnan AA, Carrière 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  CAS  Google Scholar 

  • Waller PJ (2006) Sustainable nematode parasite control strategies for ruminant livestock by grazing management and biological control. Anim Feed Sci Technol 126:277–289

    Article  Google Scholar 

  • Walsh TK, Donnan AA, Jackson F, Skuce P, Welstenholme 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  PubMed  Google Scholar 

  • Zhang Z, Gasser RB, Yang X, Yin F, Zhao G, Bao M, Pan B, Huang W, Wang C, Zou F, Zhou Y, Zhao J, Fang R, Hu M (2016) Two benzimidazole resistance-associated SNPs in the isotype-1 β-tubulin gene predominate in Haemonchus contortus populations from eight regions in China. Int J Parasitol Drugs Drug Resist 6:199–206

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This study was supported by the Ministry of Science and Technology, Higher Education, and Technical Vocational Training (MCTESTP) through the High Education, Science and Technology (HEST) Project—World Bank. We thank the technical assistance provided by Dr. Aida Cala, Mr. Carlos Sitoe and Mrs. Ana Paula from the Animal Sciences Directorate (DCA) of the Agricultural Research Institute of Mozambique (IIAM) and by Mr. Gedeão Macanze from the Provincial Veterinary Laboratory in Gaza. We express our sincere appreciation to the IIAM General Director, Dr. Olga Fafetine, and the DCA Director, Dr. Zacarias Massango, for logistical support during the field work. The technical and logistical support provided by the staff from the Molecular and Cellular Biology Laboratory (LBCM) of the Federal University of Bahia (UFBA), Brazil, is especially acknowledged.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Alsácia Atanásio-Nhacumbe.

Ethics declarations

All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. Approval from the Ethical Commission on the Use of Animals at the School of Veterinary Medicine—Federal University of Bahia (UFBA), Brazil, has been registered under EMZV-UFBA No. 09/2017.

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Section Editor: Georg von Samson-Himmelstjerna

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Atanásio-Nhacumbe, A., Lambert, S.M., da Silva Souza, B.M.P. et al. Molecular detection of benzimidazole resistance levels associated with F167Y and F200Y polymorphisms in Haemonchus contortus of goats from Mozambique. Parasitol Res 118, 245–253 (2019). https://doi.org/10.1007/s00436-018-6162-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00436-018-6162-6

Keywords

Navigation