Advanced SearchSearch Tips
Diagnosis of Benzimidazole Resistance in Haemonchus contortus of Sheep by Allele Specific PCR
facebook(new window)  Pirnt(new window) E-mail(new window) Excel Download
 Title & Authors
Diagnosis of Benzimidazole Resistance in Haemonchus contortus of Sheep by Allele Specific PCR
Tiwari, J.; Kolte, A.P.; Kumar, S.; Swarnkar, C.P.; Singh, D.; Pathak, K.M.L.;
  PDF(new window)
The study was conducted on 162 adult male Haemonchus contortus of sheep collected from Avikanagar, Jaipur and Bikaner regions to diagnose the benzimidazole (BZ) resistance in H. contortus. The BZ resistance is primarily linked with the mutation in -tubulin isotype 1 gene which substitute phenylalanine (Phe) into tyrosine (Tyr) at the 200 codon of the gene. An allele specific polymerase chain reaction (AS-PCR) technique was used for diagnosis of BZ resistance in H. contortus. In AS-PCR, one reverse primer (TGG 312) was used in two separate reactions with each of 2 forward primers (resistant TGG 331 and susceptible CAW 106 primer) that differed only at 3' nucleotide position. Therefore, the amplified products from resistant and susceptible parasites were produced 267 and 266 bp, respectively. A total of 162 parasites were genotyped, of which 130 parasites found homozygous resistant 'rr', 22 heterozygous 'rS' and 10 homozygous susceptible 'SS' type. The prevalence of 'rr' individuals was higher in Jaipur (98%) followed by Avikanagar (93%) and Bikaner (50%) regions. Overall, the prevalence of BZ resistant allele (r) was higher (87%) as compared to 13% of BZ susceptible allele (S).
Haemonchus contortus;Benzimidazole Resistance; Tubulin;AS-PCR;
 Cited by
Influence of sustained deworming pressure on the anthelmintic resistance status in strongyles of sheep under field conditions, Tropical Animal Health and Production, 2016, 48, 7, 1455  crossref(new windwow)
Alvarej-Sanchez, M. A., J. Perez-Garcia, M. A. Cruz-Rojo and F. A. Rojo-Vazquez. 2004. Real time PCR for the diagnosis of benzimidazole resistance in trichostrongylids of sheep. Vet. Parasitol. 129:291-298. crossref(new window)

Annon. 2004. Annual Progress Report: All India Network Program on Gastrointestinal Parasitism. CSWRI, Avikanagar, India, p. 1-45.

Assan, N. and S. M. Makuza, 2005. The effect on non-genetic factors on birth weight and weaning weight in three sheep breeds of Zimbabwe. Asian-Aust. J. Anim. Sci. 18(2):151-157.

Beech, R. N., R. K. Prichard and M. E. Scott. 1994. Genetic variability of the beta-tubulin genes in benzimidazole susceptible and resistant strains of Haemonchus contortus. Genet. 138:103-110.

Coles, G. C., C. Bauer, F. H. Borgsteede, S. Geerts, T. R. Klei, M. A. Taylor and M. J. Waller. 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. crossref(new window)

Dorny, P., E. Romjali, K. Feldman, A. Barubara and V. C. Pandey. 1995. Studies on the efficacy of four anthelmintics against strongyle infections of sheep in North Sumatra, Indonesia. Asian-Aust. J. Anim. Sci. 8:347-352.

Elard, L., J. Cabaret and J. F. Humbert. 1999. PCR diagnosis of benzimidazole-susceptibility or -resistance in natural populations of the small ruminant parasite, Teladorsagia circumcincta. Vet. Parasitol. 80:231-237. crossref(new window)

Elard, L., A. M. Comes and J. F. Humbert. 1996. Sequences of beta-tubulin cDNA from benzimidazole -susceptible and - resistant strains of Teladorsagia circumcincta, a nematode parasite of small ruminants. Mol. Biochem. Parasitol. 79:249-253. crossref(new window)

Elard, L. and J. F. Humbert. 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. crossref(new window)

Howlader, M. M. R., S. S. Capitan, S. L. Eduardo, N. P. Roxas and C. C. Sevilla. 1996. Effect of experimental Haemonchus contortus infection on haemoglobin concentration and packed cell volume of does. Asian-Aust. J. Anim. Sci. 9:597-601.

Howlader, M. M. R., S. S. Capitan, S. L. Eduardo and N. P. Roxas. 1997a. Effect of experimental Haemonchus contortus infection on red blood cells and white blood cells of growing goats. Asian-Aust. J. Anim. Sci. 10:679-682.

Howlader, M. M. R., S. S. Capitan, S. L. Eduardo, N. P. Roxas and C. C. Sevilla. 1997b. Performance of growing goats experimentally infected with stomach worm (Haemonchus contortus). Asian-Aust. J. Anim. Sci. 10:534-539.

Humbert, J. F., J. Cabaret, L. Elard, V. Leignel and A. Silvestre. 2001. Molecular approaches to studying benzimidazole resistance in trichostrongylid nematode parasites of small ruminants. Vet. Parasitol. 101:405-414. crossref(new window)

Jackson, F. 1993. Anthelmintic resistance- the state of play. Br. Vet. J. 149:123-138.

Kwa, M. S., F. N. Kooyman, J. H. Boersema and M. H. Roos. 1993. Effect of selection for benzimidazoles resistance in Haemonchus contortus on beta tubulin isotype 1 and isotype 2 genes. Biochem. Biophys. Res. Commun. 191:413-419. crossref(new window)

Kwa, M. S., J. G. Veenstra and M. H. Roos. 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. crossref(new window)

Le Jambre, L. F. 1976. Egg hatch as an in vitro assay of thiabendazole resistance in nematodes. Vet. Parasitol. 2:385-391. crossref(new window)

Lehrer, S., H. Davey, T. Watson and R. J. Wilkins. 1995. Sensitive PCR for detecting benzimidazole resistant sub populations of ovine nematodes in the Waikato. In Proceedings of the New Zealand Society of Animal Production, 55:209-210.

Mandal, A., K. P. Pant, D. R. Notter, P. K. Rout, R. Roy, N. K. Sinha and N. Sharma. 2005. Studies on inbreeding and its effects on growth and fleece traits of Muzaffarnagri sheep. Asian-Aust. J. Anim. Sci. 18(10):1363-1367.

Martin, P. J., N. Anderson and R. G. Jarrett. 1989. Detecting benzimidazole resistance with faecal egg count reduction tests and in vitro assays. Aust. Vet. J. 66:236-240. crossref(new window)

Pape, M., J. Posedi, K. Failing, T. Schnieder and G. Von Samson- Himmelstjerna. 2003. Analysis of the beta-tubulin codon 200 genotype distribution in a benzimidazole- susceptible and resistant cyathostome population. Parasitol. 127:53-59. crossref(new window)

Pierce, B. A. 2003. Genetics, A Conceptual Approach. W.H. Freeman and Company, NY, USA, pp. 671-672.

Roos, M. H., M. S. Kwa and W. N. Grant. 1995. New genetic and practical implications of selection for anthelmintic resistance in parasitic nematodes. Parasitol. Today. 11:148-150. crossref(new window)

Silvestre, A. and J. F. Humbert. 2000. A molecular tool for species identification and benzimidazole resistance diagnosis in larval communities of small ruminant parasites. Exp. Parasitol. 95:271-276. crossref(new window)

Silvestre, A. and J. F. Humbert. 2002. Diversity of benzimidazole resistance alleles in populations of small ruminants parasites. Int. J. Parasitol. 32:921-928. crossref(new window)

Singh, D., R. Gulyani and V. Bhasin. 1992. Occurrence of thiabendazole resistant strains of Haemonchus contortus in sheep. Ind. Vet. Med. J. 16:139-141.

Singh, D., P. K. Sanyal, C. P. Swarnkar, F. A. Khan and P. S. K. Bhagwan. 1999. Influence of diet type and pre-treatment fasting on the disposition kinetics of albendazole in sheep. Vet. Res. Commun. 23:229-240. crossref(new window)

Singh, D., C. P. Swarnkar and F. A. Khan. 2002. Anthelmintic resistance in gastrointestinal nematodes of livestock in India. J. Vet. Parasitol. 16:115-130.

Singh, D., C. P. Swarnkar, F. A. Khan, C. P. Srivastava and P. S. K. Bhagwan. 1995. Resistance to albendazole in gastrointestinal nematodes of sheep. J. Vet. Parasitol. 9:95-98.

Singh, D., C. P. Swarnkar, C. P. Srivastava, P. S. K. Bhagwan and U. Dimri. 1996. Haemonchus contortus resistant to rafoxanide in sheep. J. Vet. Parasitol. 10:53-56.

Snedecor, G. W. and W. G. Cochran. 1967. Statistical Methods. Oxfords and IBH publishing company, Culcatta, India.

Soulsby, E. J. L. 1965. Textbook of veterinary clinical parasitology, Vol. 1, helminths. Blackwell Scientific Publication. Great Britain, pp. 297-310.

Swarnkar, C. P., F. A. Khan, D. Singh and P. S. K. Bhagwan. 1999. Further studies on anthelmintic resistance in sheep at an organized farm in arid region of Rajasthan. Vet. Parasitol. 82:81-84. crossref(new window)

Swarnkar, C. P., P. K. Sanyal, D. Singh, F. A. Khan and P. S. K. Bhagwan. 2001. Anthelmintic resistance on an organized sheep farm in India. Trop. Anim. Health. Prod. 33:305-312. crossref(new window)

Swarnkar, C. P., D. Singh, F. A. Khan, R. Tiwari and S. C. Dubey. 2004. Prevalence of anthelmintic resistance and seasonal variation in efficacy of anthelmintics in sheep. In XV National Congress of Indian Association for the Advancement of Veterinary Parasitology, GBPUAT, Pantnagar, India, p. 72.

Varshney, T. R. and Y. P. Singh. 1976. A note on development of resistance of Haemonchus contortus worms against phenothiazine and thiabendazole in sheep. Indian J. Anim. Sci. 46:666-668.

Von Samson-Himmelstjerna, G., M. Pape, C. Von Witzendorff and T. Schnieder. 2002a. Allele specific PCR for the beta tubulin codon 200 TTC/TAC polymorphism using single adult and larval small strongyle (Cyathostominae) stages. J. Parasitol. 88:254-257.

Von Samson-Himmelstjerna, G., C. Von Witzendorff, G. Sievers and T. Schnieder. 2002b. Comparative use of faecal egg count reduction test, egg hatch assay and beta tubulin codon 200 genotyping in small strongyles (Cyathostominae) before and after benzimidazole treatment. Vet. Parasitol. 108:227-235. crossref(new window)

Waller, P. J. 1994. The development of anthelmintic resistance in ruminant livestock. Acta Tropica 56:233-243. crossref(new window)

Wheeler, I. E., S. J. Kendall, J. Butters, D. W. Hollomon and L. Hall. 1995. Using allele specific oligonucleotide probes to characterize benzimidazole resistance in Rhynchosporium secalis. Pestic. Sci. 43:201-209. crossref(new window)

Winterrowd, C. A., W. E. Pomroy, N. C. Sangster, S. S. Johnson and T. G. Geary. 2003. Benzimidazole resistant beta tubulin alleles in a population of parasitic nematode (Cooperia oncophora) of cattle. Vet. Parasitol. 117:161-172. crossref(new window)

Yadav, C. L., R. P. Uppal and S. Kalra. 1993. An outbreak of haemonchosis associated with anthelmintic resistance in sheep. J. Vet. Parasitol. 23:411-413.