Vet World   Vol.18   December-2025  Article - 21 

Research Article

Veterinary World, 18(12): 3959-3967

https://doi.org/10.14202/vetworld.2025.3959-3967

First molecular insight into Ovar-DRB1 exon 2 in Edilbay sheep: High heterozygosity and detection of novel variants

Saida N. Marzanova1 ORCID, Davud A. Devrishov1 ORCID, Vladislav A. Zuchkov2 ORCID, Nurbiy S. Marzanov3 ORCID, and Elizaveta A. Nikolaeva2 ORCID

1. Department of Immunology and Biotechnology, Moscow State Academy of Veterinary Medicine and Biotechnology named after K. I. Skryabin, 109472, Akademika Skryabina str., 23, Moscow, Russia.

2. Center for Biotechnology and Applied Immunology, Moscow State Academy of Veterinary Medicine and Biotechnology named after K. I. Skryabin, 109472, Akademika Skryabina str., 23, Moscow, Russia.

3. Laboratory of Molecular Genetics Farm Animals, Federal Research Center for Animal Husbandry named after Academy Member L.K. Ernst, 142132, Dubrovitsy 60, Podolsk Municipal District, Moscow Region, Russia.

Background and Aim: The Ovar-DRB1 gene, a key component of the sheep main histocompatibility complex (MHC) class II region, plays a critical role in antigen presentation and immune responsiveness. Despite the well-documented hypervariability of exon 2 in many sheep breeds, no study has yet examined DRB1 allelic composition in Edilbay sheep, a Kazakh breed highly adapted to harsh continental steppe conditions. This study aimed to characterize the diversity of Ovar-DRB1 exon 2 alleles in Edilbay sheep and to identify novel allelic variants using Sanger sequencing. 

Materials and Methods: Blood samples from 50 Edilbay sheep reared at a breeding farm in Kazakhstan were subjected to DNA extraction and DRB1 exon 2 amplification using validated primers. Polymerase chain reaction products were purified and sequenced using Sanger sequencing. Allele identification was performed through pairwise sequence alignment in SnapGene and reference comparison with the Immuno Polymorphism Database of Major Histocompatibility Complex (IPD-MHC) database. Ambiguous chromatograms and overlapping nucleotide peaks were assessed for potential novel allelic patterns. Genetic diversity indices (Ho, He, Ne, and Shannon’s H’) were calculated. 

Results: A total of 25 known DRB1 alleles were identified in the Edilbay sheep population. Genetic diversity parameters demonstrated extremely high immunogenetic variation, with observed heterozygosity (Ho) of 0.94, expected heterozygosity (He) of 0.90, an effective number of alleles (Ne) of 16.7, and Shannon’s index (H’) of 3. Several chromatograms showed overlapping peaks or substitution patterns inconsistent with known alleles, including variations at positions 243–244 and multiple additional polymorphic sites. These patterns indicate the presence of putative novel alleles that could not be unambiguously assigned by direct Sanger sequencing. Approximately 20% of samples contained undocumented variants or low-quality chromatograms requiring further resolution. 

Conclusion: This study presents the first comprehensive molecular characterization of Ovar-DRB1 exon 2 in Edilbay sheep, revealing exceptionally high genetic diversity and strong evidence for previously undescribed alleles. These findings broaden the catalog of DRB1 variants and highlight the breed’s adaptive immunogenetic potential. Further investigations using allele-specific amplification, cloning, or next-generation sequencing are recommended to precisely identify novel variants and explore associations with disease resistance and environmental adaptation. 

Keywords: Edilbay sheep, genetic diversity, immunogenetics, Kazakh sheep breed, major histocompatibility complex, novel alleles, Ovar-DRB1 exon 2, polymorphism, Sanger sequencing, sheep immune genetics.

How to cite this article: Marzanova SN, Devrishov DA, Zuchkov VA, Marzanov NS, and Nikolaeva EA (2025) First molecular insight into Ovar-DRB1 exon 2 in Edilbay sheep: High heterozygosity and detection of novel variants, Veterinary World, 18(12): 3959–3967.

Received: 12-08-2025   Accepted: 19-11-2025   Published online: 18-12-2025

Corresponding author: Saida Marzanova    E-mail: s.marzanova@mail.ru

DOI: 10.14202/vetworld.2025.3959-3967

Copyright: Marzanova, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.