Analysis of the beta-tubulin codon 200 genotype distribution in a benzimidazole-susceptible and -resistant cyathostome population.
Abstract: To study the prevalence of the polymorphism in position 200 of the beta-tubulin gene in the mechanism of benzimidazole (BZ) resistance in cyathostomes of horses, an allele-specific PCR was used to detect the genotype of individuals of BZ-susceptible and BZ-resistant populations. The molecular analysis of 100 adults recovered from an anthelmintic-naïve horse revealed 80% homozygous TTC/TTC individuals, 17% heterozygous TTC/TAC and 3% homozygous TAC/TAC. A naturally infected horse was treated with increasing fenbendazole (FBZ) dosages to select a BZ-resistant population of cyathostomes. The PCR based analysis of 3rd-stage larvae (L3) during the experiment revealed a decrease of the homozygous TTC/TTC genotype and an increase in heterozygous TTC/TAC and homozygous TAC/TAC individuals. After treatment 42.3% of the adults (n=104) were homozygous TTC/TTC, 55.8% were heterozygous TTC/TAC and only 1.9% showed the homozygous genotype TAC/TAC. The results of the molecular analysis lead to the proposal that polymorphism within codon 200 is not the only reason for the development of BZ resistance in small strongyles.
Publication Date: 2003-07-30 PubMed ID: 12885188DOI: 10.1017/s0031182003003317Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research investigated the role of polymorphism at the 200th position in the beta-tubulin gene in benzimidazole resistance in cyathostomes of horses, revealing that it may not be the only factor involved in resistance development.
Research Purpose and Method
- The study aimed to explore the prevalence of polymorphism at position 200 of the beta-tubulin gene in benzimidazole, a type of anthelmintic drug resistance among cyathostomes, a type of parasitic worm common in horses.
- Researchers employed an allele-specific Polymerase Chain Reaction (PCR) assay to determine the genotypes of both benzimidazole-susceptible and -resistant populations.
- The populations were obtained from 100 adult cyathostomes and 3rd-stage larvae (L3) retrieved from horses.
Research Findings
- Out of 100 cyathostomes, the molecular analysis revealed 80% were homozygous (TTC/TTC genotype), while 17% were heterozygous (TTC/TAC genotype), and 3% were homozygous (TAC/TAC genotype).
- These horses were anthelmintic-naïve, meaning they had not been exposed to deworming drugs before.
- Then, the researchers treated a naturally infected horse with increasing fenbendazole dosages to select a benzimidazole-resistant population of cyathostomes.
- During this phase, the researchers noted a decrease in the homozygous TTC/TTC genotype and an increase in both the heterozygous TTC/TAC and homozygous TAC/TAC.
Implications of the Research
- Based on the results, the authors concluded that polymorphism within the 200th codon of the beta-tubulin gene is not the only cause of benzimidazole resistance in small strongyles.
- This suggests that other factors might be contributing to the resistance seen in these parasitic worms, highlighting the need for further investigation.
Cite This Article
APA
Pape M, Posedi J, Failing K, Schnieder T, von Samson-Himmelstjerna G.
(2003).
Analysis of the beta-tubulin codon 200 genotype distribution in a benzimidazole-susceptible and -resistant cyathostome population.
Parasitology, 127(Pt 1), 53-59.
https://doi.org/10.1017/s0031182003003317 Publication
Researcher Affiliations
- Institute of Parasitology, School of Veterinary Medicine Hannover, Buenteweg 17, D-30559 Hannover, Germany. Michaela.Pape@tiho-hannover.de
MeSH Terms
- Alleles
- Animals
- Anthelmintics / pharmacology
- Benzimidazoles / pharmacology
- Codon / analysis
- DNA, Helminth / genetics
- Drug Resistance
- Genotype
- Horses
- Molecular Weight
- Strongyle Infections, Equine / diagnosis
- Strongyle Infections, Equine / epidemiology
- Strongyle Infections, Equine / parasitology
- Strongyloidea / genetics
- Strongyloidea / isolation & purification
- Tubulin / analysis
- Tubulin / genetics
Citations
This article has been cited 13 times.- Onder Z, Yildirim A, Duzlu O, Ciloglu A, Yetismis G, Karabulut F, Inci A. Detection of SNPs and benzimidazole resistance in strongyle nematode eggs of horses by allele-specific PCR. Parasitol Res 2023 Sep;122(9):2037-2043.
- Cain JL, Nielsen MK. The equine ascarids: resuscitating historic model organisms for modern purposes. Parasitol Res 2022 Oct;121(10):2775-2791.
- Özben M, von Samson-Himmelstjerna G, Freiin von Streit MKB, Wilkes EJA, Hughes KJ, Krücken J. Absence of Polymorphisms in Codons 167, 198 and 200 of All Seven β-Tubulin Isotypes of Benzimidazole Susceptible and Resistant Parascaris spp. Specimens from Australia. Pathogens 2022 Apr 20;11(5).
- Roose S, Avramenko RW, Pollo SMJ, Wasmuth JD, Ame S, Ayana M, Betson M, Cools P, Dana D, Jones BP, Mekonnen Z, Morosetti A, Venkatesan A, Vlaminck J, Workentine ML, Levecke B, Gilleard JS, Geldhof P. Characterization of the β-tubulin gene family in Ascaris lumbricoides and Ascaris suum and its implication for the molecular detection of benzimidazole resistance. PLoS Negl Trop Dis 2021 Sep;15(9):e0009777.
- Arsenopoulos KV, Fthenakis GC, Katsarou EI, Papadopoulos E. Haemonchosis: A Challenging Parasitic Infection of Sheep and Goats. Animals (Basel) 2021 Feb 1;11(2).
- Dixit AK, Das G, Dixit P, Singh AP, Kumbhakar NK, Sankar M, Sharma RL. An assessment of benzimidazole resistance against caprine nematodes in Central India. Trop Anim Health Prod 2017 Oct;49(7):1471-1478.
- Vijayasarathi MK, Sreekumar C, Venkataramanan R, Raman M. Influence of sustained deworming pressure on the anthelmintic resistance status in strongyles of sheep under field conditions. Trop Anim Health Prod 2016 Oct;48(7):1455-62.
- Shokrani H, Shayan P, Eslami A, Nabavi R. Benzimidazole -Resistance in Haemonchus contortus: New PCR-RFLP Method for the Detection of Point Mutation at Codon 167 of Isotype 1 β-Tubulin Gene. Iran J Parasitol 2012;7(4):41-8.
- Lake SL, Matthews JB, Kaplan RM, Hodgkinson JE. Determination of genomic DNA sequences for beta-tubulin isotype 1 from multiple species of cyathostomin and detection of resistance alleles in third-stage larvae from horses with naturally acquired infections. Parasit Vectors 2009 Sep 25;2 Suppl 2(Suppl 2):S6.
- Diawara A, Drake LJ, Suswillo RR, Kihara J, Bundy DA, Scott ME, Halpenny C, Stothard JR, Prichard RK. Assays to detect beta-tubulin codon 200 polymorphism in Trichuris trichiura and Ascaris lumbricoides. PLoS Negl Trop Dis 2009;3(3):e397.
- Shayan P, Eslami A, Borji H. Innovative restriction site created PCR-RFLP for detection of benzimidazole resistance in Teladorsagia circumcincta. Parasitol Res 2007 Apr;100(5):1063-8.
- Blackhall WJ, Drogemuller M, Schnieder T, von Samson-Himmelstjerna G. Expression of recombinant beta-tubulin alleles from Cylicocyclus nassatus (Cyathostominae). Parasitol Res 2006 Nov;99(6):687-93.
- Jamshidpour R, Nabavi R, Moadab H, Rezaie F, Chale AC, Sargison N. Evaluation of Benzimidazole Resistance in Equine Cyathostomins in the Kermanshah Province of Iran Using Coprological Analysis and Allele-Specific PCR. Iran J Parasitol 2025 Jul-Sep;20(3):389-399.
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