Molecular detection of benzimidazole resistance levels associated with F167Y and F200Y polymorphisms in Haemonchus contortus of goats from Mozambique
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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.
KeywordsAnthelmintic resistance β-Tubulin Gastrointestinal nematodes Real-time PCR Small ruminants
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.
Compliance with ethical standards
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.
- 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 69:215–220Google 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 9:313–319Google 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 186:344–349CrossRefGoogle 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 9:349–360CrossRefGoogle 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 1–2Google 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 13:71–78CrossRefGoogle 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 81–112Google 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 8:28–34Google Scholar
- Melo ACFL, Bevilaqua CML (2005) Genetic approach of anthelmintic resistance in Haemonchus contortus: a review article. Revista Portuguesa de Ciências Veterinárias 100:141–146Google 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
- 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. 6:230–240CrossRefGoogle Scholar
- Salgado JA, Santos CP (2016) Overview of anthelmintic resistance of gastrointestinal nematodes of small ruminants in Brazil. Rev Brasileira Parasitol Vet 25Google Scholar
- Ueno H, Gonçalves PC (1998) Handbook for diagnosis of helminthoses of ruminants, 4th edn. Japan International Cooperation, Tokyo, p 143Google 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 6:199–206CrossRefGoogle Scholar