Veterinary Research Forum

Assessment of DNA integrity in bovine viral diarrhea virus-infected cells using alkaline single-cell gel electrophoresis

Document Type : Original Article

Author

Aidin Rahim Tayefeh

Department of Animal Biotechnology, Faculty of Agricultural Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran

https://doi.org/10.30466/vrf.2025.2058533.4732
Abstract
Bovine viral diarrhea virus is a pestivirus of the Flaviviridae family including two biotypes, cytopathic (CP) and non-CP (NCP). This study aimed to evaluate DNA damage and apoptosis in Madin-Darby bovine kidney cells following infection with both biotypes. The MDBK monolayers were inoculated with a final dose of virus (1.00 × 10³ Tissue Culture Infectious Dose 50% mL-1) and incubated for 24 hr. DNA strand integrity was assessed using alkaline single-cell gel electrophoresis, and DNA damage was quantified through tail moment and olive tail moment indices (n = 3). Apoptosis was evaluated using annexin V–fluorescein isothiocyanate/propidium iodide flow cytometry to determine early and late apoptotic cell populations. Both biotypes significantly increased DNA fragmentation compared to the control group. The tail moment values were 15.89 ± 2.13 (control), 57.63 ± 16.20 (NCP), and 68.15 ± 9.93 (CP); while, olive tail moment values were 8.71 ± 1.01 (control), 29.35 ± 9.18 (NCP), and 35.14 ± 6.90 (CP). Apoptosis analysis showed a higher percentage of apoptotic cells in infected groups, with CP biotype of bovine viral diarrhea virus inducing the greatest early and late apoptotic responses, being consistent with its CP nature. Overall, both biotypes caused notable genomic injury and apoptosis in Madin-Darby bovine kidney cells, with CP producing the highest level of damage, confirming single-cell gel electrophoresis combined with apoptosis assays as sensitive tools for detecting virus-mediated genomic instability and supporting their potential application in breeding programs aimed at enhancing resistance to infectious diseases.

Keywords

Apoptosis
Bovine viral diarrhea virus
DNA integrity
Madin-Darby bovine kidney cells
Single cell gel electrophoresis
Subjects
1.     Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol 2007; 35(4): 495-516.
2.     Khodakaram-Tafti A, Farjanikish GH. Persistent bovine viral diarrhea virus (BVDV) infection in cattle herds. Iran J Vet Res 2017; 18(3): 154-163.
3.     Oguejiofor CF, Thomas C, Cheng Z, et al. Mechanisms linking bovine viral diarrhea virus (BVDV) infection with infertility in cattle. Anim Health Res Rev 2019; 20(1): 72-85.
4.     Kępka K, Wójcik E, Wysokińska A. Identification of genomic instability in cows infected with BVD virus. Animals (Basel) 2023; 13(24): 3800. doi: 10.3390/ ani13243800.
5.     Harutyunyan T, Sargsyan A, Kalashyan L, et al. DNA damage in moderate and severe COVID-19 cases: relation to demographic, clinical, and laboratory parameters. Int J Mol Sci 2024; 25(19): 10293. doi: 10.3390/ijms251910293.
6.     Roulston A, Marcellus RC, Branton PE. Viruses and apoptosis. Annu Rev Microbiol 1999; 53: 577-628.
7.     Rieger AM, Nelson KL, Konowalchuk JD, et al. Modified annexin V. propidium iodide apoptosis assay for accurate assessment of cell death. J Vis Exp 2011; (50): 2597. doi: 10.3791/2597.
8.     Singh NP, McCoy MT, Tice RR, et al. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 1988; 175(1): 184-191.
9.     Reed LJ. A simple method of estimating fifty per cent endpoints. Am J Epidemiol 1938; 27: 493-497.
10. Kobayashi H, Sugiyama C, Morikawa Y, et al. A comparison between manual microscopic analysis and computerized image analysis in the single cell gel electrophoresis assay. MMS Commun 1995; 3(2):103-115.
11. Dzitsiuk V, Tipilo H. Chromosomal anomalies in dairy cattle as reasons of impaired fertility. Agric Sci Pract 2019; 1: 60-66.
12. Jones KM, Bryan A, McCunn E, et al. The causes and consequences of DNA damage and chromosomal instability induced by human papillomavirus. Cancers (Basel) 2024; 16(9): 1662. doi: 10.3390/cancers 16091662.
13. Abrahams RR, Majumder K. Small genomes, big disruptions: parvoviruses and the DNA damage response. Viruses 2025; 17(4): 494. doi: 10.3390/ v17040494.
14. Abiri E, Mirzaii M, Moghbeli M, et al. Investigating DNA damage caused by COVID-19 and influenza in post COVID-19. Mamm Genome 2025; 36(1): 200-212.
15. Çakır DA, Yirün A, Erdemli-Köse SB, et al. The combined effects of HSV-1 glycoprotein D and aluminum hydroxide on human neuroblastoma cells: insights into oxidative DNA damage, apoptosis, and epigenetic modifications. Neurotoxicology 2025; 108: 123-133.
16. Mihaljevic O, Zivancevic-Simonovic S, Cupurdija V, et al. DNA damage in peripheral blood lymphocytes of severely ill COVID-19 patients in relation to inflammatory markers and parameters of hemostasis. Mutagenesis 2022; 37(3-4): 203-212.
17. Kaleelullah RA, Garugula N. Teratogenic genesis in fetal malformations. Cureus 2021; 13(2): e13149. doi: 10.7759/cureus.13149.
Veterinary Research Forum
Volume 17, Issue 4
April 2026
Pages 267-272

  • Receive Date 21 April 2025
  • Revise Date 22 November 2025
  • Accept Date 10 December 2025

Home

Guide for Authors

Submit Manuscript

Reviewers

Contact Us