Asian-Australasian Journal of Animal Sciences

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
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Diagnosis of Benzimidazole Resistance in Haemonchus contortus of Sheep by Allele Specific PCR

  • Tiwari, J. (Department of Veterinary Parasitology, Rajasthan Agricultural University) ;
  • Kolte, A.P. (Animal Biotechnology Section, Central Sheep and Wool Research Institute) ;
  • Kumar, S. (Animal Biotechnology Section, Central Sheep and Wool Research Institute) ;
  • Swarnkar, C.P. (Division of Animal Health, Central Sheep and Wool Research Institute, Avikanagar) ;
  • Singh, D. (Division of Animal Health, Central Sheep and Wool Research Institute, Avikanagar) ;
  • Pathak, K.M.L. (Department of Veterinary Parasitology, Rajasthan Agricultural University)
  • Received : 2005.09.21
  • Accepted : 2006.02.01
  • Published : 2007.01.01
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Abstract

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 ${\beta}$-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).

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References

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