Research Article | 10 Jul 2026

Leucaena leucocephala fermented with Lactiplantibacillus plantarum TISTR1284 as a sustainable fishmeal substitute enhances immune responses and resistance to Aeromonas hydrophila infection in giant freshwater prawns (Macrobrachium rosenbergii)

Arnon Pudgerd^1,2

, Laorrat Phuapittayalert^1,2

, Kamonwan Jongsomchai^1,2

, Sirilak Sanpa^2,3

, Sirikarn Sanpa^2,3

, Paiboon Panase^4,5

, Pornpan Pumirat⁶

, Sukanya Saedan^7,8

, Rapeepun Vanichviriyakit^7,8

, and Charoonroj Chotwiwatthanakun^8,9

1. Division of Anatomy, School of Medical Sciences, University of Phayao, Maeka, Muang, Phayao 56000, Thailand.

2. Unit of Excellence in Plant and Aquaculture, University of Phayao, Maeka, Muang, Phayao 56000, Thailand.

3. Division of Microbiology, School of Medical Sciences, University of Phayao, Maeka, Muang, Phayao 56000, Thailand.

4. Division of Fisheries, School of Agriculture and Natural Resources, University of Phayao, Phayao 56000, Thailand.

5. Unit of Excellence “Physiology for the Sustainable Production and Management of Aquatic Animals” Division of Fisheries, School of Agriculture and Natural Resources, University of Phayao, Phayao 56000, Thailand.

6. Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand.

7. Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Bangkok 10400, Thailand.

8. Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Mahidol University, Rama VI Road, Bangkok 10400, Thailand.

9. Nakhonsawan Campus, Mahidol University, Nakhonsawan 60130, Thailand.

Corresponding author: Charoonroj Chotwiwatthanakun (charoonroj.cho@mahidol.ac.th)

Received: 26-02-2026 , Accepted: 17-06-2026 , Published: 10-07-2026

VETERINARY WORLD | Article No. 13 | pg no. 2876-2898 | Vol. 19, Issue 7 | DOI: 10.14202/vetworld.2026.2876-2898

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ABSTRACT

Background and Aim: The increasing cost and limited availability of fishmeal have intensified the search for sustainable alternative protein sources in aquaculture. Fermentation of plant-based feed ingredients can improve their nutritional value and functional properties. This study evaluated the effects of dietary inclusion of Leucaena leucocephala leaves fermented with Lactiplantibacillus plantarum TISTR1284 as a partial fishmeal substitute on immune responses, intestinal health, and resistance to Aeromonas hydrophila infection in giant freshwater prawns (Macrobrachium rosenbergii).

Materials and Methods: Adult M. rosenbergii were randomly assigned to three dietary treatments: a control diet and diets in which 10% or 20% of fishmeal was replaced with fermented L. leucocephala. Prawns were fed the experimental diets for 2 weeks before challenge with A. hydrophila. Total hemocyte counts, phenoloxidase activity, hematopoietic tissue proliferation, immune-related gene expression, intestinal histology, antioxidant properties of the diets, and post-challenge survival were evaluated.

Results: Fermentation enhanced the nutritional quality of L. leucocephala by increasing essential amino acid content and reducing mimosine concentration. Diets containing fermented L. leucocephala exhibited greater antioxidant activity, total phenolic content, and total flavonoid content than the control diet. Prawns receiving fermented diets showed significantly increased total hemocyte counts, enhanced hematopoietic cell proliferation, elevated crustacean hematopoietic factor expression, and increased phenoloxidase activity. The expression of key immune-related genes, including IMD, Relish, HSP70, Cu/Zn-SOD, and anti-lipopolysaccharide factor (ALF), was upregulated, whereas TRAF6 and Dorsal expression was moderated during infection. Intestinal morphology remained normal, indicating that dietary inclusion of fermented L. leucocephala did not adversely affect gut structure. Following A. hydrophila challenge, prawns fed fermented diets exhibited a 26.14% reduction in mortality compared with the control group, demonstrating improved disease resistance.

Conclusion: Partial replacement of fishmeal with L. leucocephala leaves fermented by L. plantarum TISTR1284 enhanced innate immune responses, improved antioxidant status, and increased resistance to A. hydrophila infection in M. rosenbergii without compromising intestinal health. Fermented L. leucocephala represents a promising sustainable functional feed ingredient for freshwater prawn aquaculture and may contribute to reducing dependence on fishmeal in commercial production systems.

Keywords: Aeromonas hydrophila, disease resistance, fishmeal replacement, giant freshwater prawn, immune response, Lactiplantibacillus plantarum, Leucaena leucocephala, sustainable aquaculture.