Detection of benzimidazole resistance in Haemonchus contortus using RFLP-PCR technique

Tiwari, Jawahar L.; Kumar, Sunil; Kolte, Atul P.; Swarnkar, C.P.; Singh, Deepak; Pathak, K. M. L.

doi:10.1016/j.vetpar.2006.02.003

Cited by 49 publications

(31 citation statements)

References 25 publications

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“…The resistance status of H. contortus in the four farms were almost similar. Tiwari et al (2006) reported similar genotypic frequency using RFLP-PCR in Rajasthan, India. In addition to the SNPs of b-tubulin, genetic changes in P-glycoprotein and cytochrome P-450 also contribute to the development of anthelmintic resistance (Kerboeuf et al 2003).…”

Section: Discussionmentioning

confidence: 64%

“…This has led to the development of multiple drug resistant (MDR) strains of H. contortus in many parts of the world viz., South America (Nari et al 1996), South Africa (van Wyk et al 1999, Malaysia (Chandrawathani et al 2003), Southeast USA (Mortensen et al 2003), Australia (Besier and Love 2003), New Zealand (Leathwick et al 2001) and India (Gill 1996;Swarnkar et al 1999;Easwaran et al 2009). In Tiwari et al (2006) reported the prevalence rate of BZ resistance in H. contortus of Indian isolates by RFLP-PCR technique. As H. contortus is an economically important parasite affecting sheep and goats in India, it is highly essential to review the benzimidazole resistance status, so that an effective ITS is one of the most variable nuclear loci and hence has been a popular choice in developing highly sensitive techniques for detection of helminths (Mochizuki et al 2006).…”

Section: Discussionmentioning

confidence: 99%

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Genotyping of benzimidazole resistant and susceptible isolates of Haemonchus contortus from sheep by allele specific PCR

et al. 2016

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Extensive and indiscriminate use of the benzimidazole class of drugs has led to the onset of anthelmintic resistance. In tropical countries like India, Haemonchus contortus is the most pathogenic parasite infecting sheep and goats. The widespread presence of resistant helminths (especially H. contortus) threatens the livestock farming. The use of various drugs has led to single nucleotide polymorphism that causes specific amino acid substitutions in b-tubulin protein of H. contortus to confer resistance. This emphasizes the need for a survey on the present status of resistance in India. In this study, allele specific PCR was employed to screen the presence of a SNP, a thymine-toadenine transversion which leads to substitution of amino acid in codon 200 of b-tubulin gene that is correlated specifically with BZ resistance. Third stage larvae (L3) from pooled faecal cultures of four organized sheep farms served as a source of genomic DNA for identification of H. contortus and further genotype analysis. A total of 1000 larvae was screened, out of which 673 larvae were identified as H. contortus. Among 673 H. contortus larvae, 539 larvae (80 %) were genotyped as homozygous resistant (rr) and remaining 134 (20 %) were heterozygous susceptible (Sr) by allele specific PCR. The concluded resistance status reasons out the failure of anthelmintic drug in treating ruminants. Immediate steps are needed to avoid further aggravation of the problem. Target selective treatment by reviewing the resistance status of individual drugs, appropriate use of anthelmintic drugs and other control strategies will provide a pragmatic option for delaying the further spread of anthelmintic resistance.

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Section: Discussionmentioning

confidence: 64%

Section: Discussionmentioning

confidence: 99%

Genotyping of benzimidazole resistant and susceptible isolates of Haemonchus contortus from sheep by allele specific PCR

et al. 2016

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“…Moreover, only homozygous ('rr') genotypes appeared to survive with the recommended dose of treatment (ELARD et al, 1999;HUMBERT et al, 2001). In contrast, TIWARI et al (2006) reported the high frequency of resistant genotypes in H. contortus collected from different regions of Rajasthan, considering 'rS' as resistant genotypes.…”

Section: Discussionmentioning

confidence: 90%

“…In India, the occurrence of BZ resistance in H. contortus has been reported in different sheep populations since 1976 by conventional (VARSHNEY and SINGH, 1976) and molecular techniques (TIWARI et al, 2006;KAUSHIK et al, 2016). Among the molecular assays, PCR-RFLP has been found to be an economically viable genotyping assay and to detect the SNP's at codon 167 and 200 of the β-tubulin isotype 1 gene of H. contortus (SHOKRANI et al, 2012).…”

mentioning

confidence: 99%

Variation in the β-tubulin isotype 1 gene at codon 167 and 200 responsible for benzimidazole resistance in Haemonchus contortus in sheep of Krishna District, AP, India

et al. 2018

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“…Protocolos de PCR convencional e em tempo real (rt-PCR) foram analisados para a detecção de SNPs nos códons 167 e 200 na β-tubulina (Samson--Himmelstjerna 2006). Para a determinação da resistência associada ao códon TAC na posição 200, técnicas de PCR--RFLP (restriction fragment length polymorphism ou polimorfismo de comprimento de fragmentos de restrição) foram desenvolvidas para H. contortus (Tiwari et al 2006) e T. circumcincta (Shayan et al 2007). A técnica de pirosequenciamento também se mostrou rápida e adequada para a detecção múltipla de SNPs (Samson-Himmelstjerna et al 2007).…”

Section: Técnicas Moleculares Para O Diagnóstico Da Resistênciaunclassified

Resistência anti-helmíntica em nematoides gastrintestinais de pequenos ruminantes: avanços e limitações para seu diagnóstico

Fortes

Molento

2013

Pesq. Vet. Bras.

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A seleção e a crescente disseminação de nematoides resistentes aos anti-helmínticos mais comumente utilizados, benzimidazóis (BZs), imidazotiazóis e lactonas macrocíclicas (LMs), constituem um sério entrave na produção de pequenos ruminantes em todo o mundo. O uso de métodos eficientes e sensíveis para a detecção e o monitoramento da resistência anti-helmíntica no campo torna-se urgente, especialmente para os grupos de BZs e LMs, devido aos constantes relatos de resistência. A obtenção de um diagnóstico preciso e precoce da resistência é extremamente importante para auxiliar a tomada de decisão em programas de controle parasitário, com o objetivo de preservar a vida útil dos produtos e limitar o desenvolvimento da resistência nas populações de nematoides. Os testes in vivo e, mais recentemente, os testes in vitro têm sido desenvolvidos para a detecção de nematoides resistentes aos principais grupos de anti-helmínticos. No entanto, a disponibilidade de testes in vitro validados e o seu uso prático ainda são muito limitados. Embora o teste de redução na contagem de ovos nas fezes (TRCOF, in vivo - indireto) seja o principal método de escolha para a detecção de resistência no campo, vem recebendo críticas quanto à validade dos resultados, e passa por significativas modificações. Além disso, o desenvolvimento de técnicas moleculares a partir de alterações genômicas gerou avanços consideráveis nessa área de investigação, com o uso de mutações nos códons 167, 198 e 200 do gene da β-tubulina como principais SNPs (polimorfismos de nucleotídeo único; do inglês Single Nucleotide Polymorphisms) associados à resistência aos BZs. A presente revisão tem o objetivo de discutir os métodos de diagnóstico disponíveis para a detecção de resistência anti-helmíntica em nematoides de pequenos ruminantes, destacando progressos e obstáculos para seu uso na rotina laboratorial e no campo.

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Detection of benzimidazole resistance in Haemonchus contortus using RFLP-PCR technique

Cited by 49 publications

References 25 publications

Genotyping of benzimidazole resistant and susceptible isolates of Haemonchus contortus from sheep by allele specific PCR

Genotyping of benzimidazole resistant and susceptible isolates of Haemonchus contortus from sheep by allele specific PCR

Variation in the β-tubulin isotype 1 gene at codon 167 and 200 responsible for benzimidazole resistance in Haemonchus contortus in sheep of Krishna District, AP, India

Resistência anti-helmíntica em nematoides gastrintestinais de pequenos ruminantes: avanços e limitações para seu diagnóstico

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