Cost-effective medium optimization and functional enhancement of Lactiplantibacillus plantarum 22F for industrial-scale probiotic production in swine feed

Vet World Vol.18 June-2025 Article - 33

Research Article

Veterinary World, 18(6): 1759-1776

https://doi.org/10.14202/vetworld.2025.1759-1776

Nay Zin Myo^1,2, Ratchnida Kamwa^1,2, Benjamas Khurajog¹, Pawiya Pupa³, Wandee Sirichokchatchawan^4,5, David J. Hampson⁶, and Nuvee Prapasarakul^1,4

1. Department of Microbiology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand.

2. The International Graduate Course of Veterinary Science and Technology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand.

3. Department of Biology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.

4. Center of Excellence in Diagnosis and Monitoring Animal Pathogens, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand.

5. College of Public Health Sciences, Chulalongkorn University, Bangkok 10330, Thailand.

6. School of Veterinary Medicine, Murdoch University, Perth, Western Australia, 6150, Australia.

Background and Aim: Industrial-scale probiotic production requires economically viable media formulations that do not compromise strain functionality. This study aimed to develop a cost-effective medium for cultivating Lactiplantibacillus plantarum 22F (L22F), a probiotic candidate isolated from swine feces, while evaluating its industrial viability and functional metabolic profile.

Materials and Methods: Carbon (glucose, sucrose, and dextrose) and nitrogen (yeast extract, soy protein isolate, and whey protein concentrate) sources were screened using one-variable-at-a-time and Plackett–Burman design, followed by Response Surface Methodology for optimization. Fermentation was scaled from a flask to 50 L fermenters at 37 °C and pH 6.50 ± 0.05. Cell viability, pH, and residual sugar were monitored. Functional assessments included stress tolerance assays (heat, acid, bile, and oxidative stress) and untargeted metabolomic profiling using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry.

Results: The optimal medium comprised 9 g/L glucose, 14.1 g/L soy protein isolate, and 14.1 g/L yeast extract, supplemented with minerals. In 50 L fermentation, L22F achieved 9.20 log colony-forming units/mL at 12 h, with residual sugar at 1.50 g/L and pH 3.99. Compared to de Man, Rogosa, and Sharpe, the modified medium reduced production cost by 70%–88%, improved fermentation efficiency, and supported enhanced stress resilience. Metabolomic analysis revealed an elevated production of bioactive metabolites, particularly 1,4-dihydroxy-2-naphthoic acid and indolelactic acid, which are known to support gut homeostasis, anti-inflammatory responses, and probiotic efficacy.

Conclusion: This study presents a cost-effective and scalable fermentation medium specifically designed for high-density L22F production. Beyond economic advantages, the medium enhanced the functional properties of L22F, supporting its application as a sustainable probiotic feed additive for swine. These findings establish a foundation for further industrial application and in vivo validation.

Keywords: cost-effective production, fermentation optimization, Lactiplantibacillus plantarum 22F, metabolomics, modified medium, stress tolerance, swine probiotic.

How to cite this article: Myo NZ, Kamwa R, Khurajog B, Pupa P, Sirichokchatchawan W, Hampson DJ, and Prapasarakul N (2025) Cost-effective medium optimization and functional enhancement of Lactiplantibacillus plantarum 22F for industrial-scale probiotic production in swine feed, Veterinary World, 18(6): 1759-1776.

Received: 21-12-2024 Accepted: 23-05-2025 Published online: 27-06-2025

Corresponding author: Nuvee Prapasarakul E-mail: nuvee.p@chula.ac.th

DOI: 10.14202/vetworld.2025.1759-1776

Copyright: Myo, 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.