Detection of SNPs and benzimidazole resistance in strongyle nematode eggs of horses by allele-specific PCR


Onder Z., Yildirim A., Duzlu Ö., Ciloglu A., Yetismis G., Karabulut F., ...More

PARASITOLOGY RESEARCH, vol.122, no.9, pp.2037-2043, 2023 (SCI-Expanded, Scopus)

  • Publication Type: Article / Article
  • Volume: 122 Issue: 9
  • Publication Date: 2023
  • Doi Number: 10.1007/s00436-023-07903-6
  • Journal Name: PARASITOLOGY RESEARCH
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, Aquatic Science & Fisheries Abstracts (ASFA), BIOSIS, CAB Abstracts, MEDLINE, Veterinary Science Database
  • Page Numbers: pp.2037-2043
  • Keywords: Strongyle nematode egg, Horse, Benzimidazole resistance, Single nucleotide polymorphism
  • Erciyes University Affiliated: Yes

Abstract

This study was conducted to determine single nucleotide polymorphisms (SNPs) and the benzimidazole (BZ) resistance in strongyle nematode egg populations in horses using molecular techniques. A total of 200 fecal samples were collected from horses in 26 farms in two provinces (Kayseri and Nevsehir) of the Central Anatolia Region of Turkiye between May and August 2022. The flotation method was used to detect strongyle nematode eggs in the fecal samples of the horses. Afterward, strongyle nematode eggs were collected, and the allele-specific polymerase chain reaction (AS-PCR) technique was used to detect the BZ resistance. BZ-susceptible and BZ-resistant PCR products were sequenced to determine single nucleotide polymorphisms (SNPs) in the & beta;-tubulin isotype 1 gene. The strongyle nematode eggs were determined in 85 (42.5%) out of 200 fecal samples. AS-PCR detected 50.58% (43/85) BZ-resistant (homozygous resistant) and 36.4% (31/85) BZ-susceptible (homozygous susceptible) genes in the strongyle eggs. Both BZ-resistant and BZ-susceptible genes (heterozygous) were determined in 11 samples. BZ-resistant and BZ-susceptible allele frequencies were determined as 57.0% (48.5/85) and 43.0% (36.5/85), respectively. SNPs were detected only in codon 200 of the & beta;-tubulin isotype 1 gene in four sequenced isolates of the two resistant and two susceptible isolates. This study is the first molecular report on BZ resistance in strongyle nematode eggs in horses in Turkiye. The widespread prevalence of BZ-resistant alleles in equine strongyle nematodes shows the requirement for the immediate usage of other anthelmintics instead of the BZ group drugs for the effective management and control of equine strongyle nematodes.