Use of a synthetic antimicrobial peptide (PA-13) alone or in combination with reduced gentamicin levels to control Escherichia coli contamination in stored boar semen

Vet World Vol.19 February-2026 Article - 23

Research Article

Veterinary World, 19(2): 771-781

https://doi.org/10.14202/vetworld.2026.771-781

Krittika Keeratikunakorn¹, Panida Chanapiwat², Ratchaneewan Aunpad³, Natharin Ngamwongsatit^4,5, and Kampon Kaeoket²

1. Department of Anatomy, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.

2. Semen Laboratory, Department of Clinical Sciences and Public Health, Faculty of Veterinary Science, Mahidol University, 999 Phuttamonthon 4 Rd., Salaya, Phuttamonthon, Nakhon Pathom 73170, Thailand.

3. Graduate Program in Biomedical Sciences, Faculty of Allied Health Sciences, Thammasat University, Rangsit Campus, Klong Luang, Pathumthani 12120, Thailand.

4. Department of Clinical Sciences and Public Health, Faculty of Veterinary Science, Mahidol University, 999 Phuttamonthon 4 Rd., Salaya, Phuttamonthon, Nakhon Pathom 73170, Thailand.

5. Laboratory of Bacteria, Veterinary Diagnostic Center, Faculty of Veterinary Science, Mahidol University, 999 Phuttamonthon 4 Rd., Salaya, Phuttamonthon, Nakhon Pathom 73170, Thailand. .

Background and Aim: Bacterial contamination during liquid storage of boar semen negatively affects sperm quality and fertility outcomes, necessitating the routine use of antibiotics in semen extenders. However, increasing concerns regarding antimicrobial resistance have encouraged the development of alternative antimicrobial strategies. Synthetic antimicrobial peptides (AMPs) have demonstrated broad-spectrum antibacterial activity and may serve as potential substitutes or adjuncts to conventional antibiotics. This study aimed to evaluate the antimicrobial efficacy of the synthetic peptide PA-13 alone and in combination with reduced gentamicin concentrations against Escherichia coli isolated from boar semen, as well as its effects on semen quality during storage at 18°C.

Materials and Methods: Two experiments were conducted. In Experiment I, fresh semen samples collected from seven healthy adult boars were diluted in Beltsville Thawing Solution supplemented with gentamicin (200 μg/mL, positive control), without antibiotics (negative control), or PA-13 at concentrations of 62.5, 31.25, and 15.625 μg/mL. Total bacterial counts were measured at 0, 24, 48, and 72 h of storage, while semen quality parameters, including sperm motility, viability, acrosomal integrity, and mitochondrial membrane potential, were evaluated on days 1, 3, and 5. In Experiment II, isolated E. coli was incubated with PA-13 alone or in combination with varying gentamicin concentrations, and bacterial growth was monitored over 24 h using optical density measurements.

Results: PA-13 effectively inhibited bacterial proliferation in extended semen during the first 24 h of storage, with lower concentrations (15.625 and 31.25 μg/mL) showing greater antibacterial activity than the higher concentration (62.5 μg/mL). Semen quality parameters were comparable among groups on day 1; however, prolonged storage demonstrated that the highest PA-13 concentration negatively affected sperm motility and viability. Lower PA-13 concentrations preserved semen quality similar to that of the gentamicin-treated control. In Experiment II, combination treatments exhibited synergistic effects, with PA-13 at 3.906 μg/mL combined with gentamicin at 100 μg/mL inhibiting E. coli growth equivalently to gentamicin at 200 μg/mL alone.

Conclusion: PA-13 effectively controls bacterial contamination in stored boar semen while maintaining semen quality at appropriate concentrations. Its combination with gentamicin enables a substantial reduction in antibiotic dosage without compromising antibacterial efficacy. These findings support the use of AMPs as alternative or complementary antimicrobial agents in semen extenders to reduce antibiotic use in the swine industry.

Keywords: artificial insemination, antimicrobial peptides, boar semen storage, Escherichia coli, gentamicin reduction, semen extenders, sperm quality, synthetic peptide PA-13.

How to cite this article: Keeratikunakorn K, Chanapiwat P, Aunpad R, Ngamwongsatit N, Kaeoket K. Use of a synthetic antimicrobial peptide (PA-13) alone or in combination with reduced gentamicin levels to control Escherichia coli contamination in stored boar semen. Vet. World, 2026; 19(2):771-781.

Received: 20-10-2025 Accepted: 23-01-2026 Published online: 26-02-2026

Corresponding author: Kampon Kaeoket E-mail: kampon.kae@mahidol.ac.th

DOI: 10.14202/vetworld.2026.771-781

Copyright: Keeratikunakorn, 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.