Anti-Vibrio Compounds from Digestive Symbiont Bacteria of Whiteleg Shrimp (Litopenaeus vannamei) from Traditional Ponds in Kaur, Bengkulu, Indonesia as Probiotic Candidates for Shrimp Aquaculture
Abstract
Kaur Regency is one of the areas with high potential for whiteleg shrimp pond cultivation, but it often faces shrimp deaths and crop failure due to infection with pathogenic Vibrio spp. and antibiotic resistance. One alternative preventive is to utilise potential symbiotic bacteria from the digestive tract that can produce anti-Vibrio and enzymatic activities. This study aimed to select potential digestive symbiont bacteria based on their anti-Vibrio ability through isolation, purification, identification, biochemical tests, and anti-Vibrio tests. Candidates showing activity were further tested for toxicity, hemolysin, effectiveness, enzymatic, and molecular identification. Out of 40 digestive symbiont isolates, two Bacillus isolates of the γ-hemolytic type, namely ALF 3 and ALF 6, showed the most potential against seven pathogenic Vibrio spp. isolates (OALF 1, OALF 2, OALF 3, OALF 5, WALF 7, WALF 8, and WALF 10) from the digestive tract and infected whiteleg shrimp pond water, with toxicity percentage of 86.67-100 %, α and β-hemolytic type pathogenicity in BA. Based on their anti-Vibrio activity, amylolytic and proteolytic abilities, ALF 3 and ALF 6 were identified as Bacillus stercoris and Bacillus amyloliquefaciens, respectively, with effectiveness percentages of 89.7-100 % for B.stercoris, 87.7-100 % for B.amyloliquefaciens, and 97.7-100 % for the consortium compared to the treatment without co-culture with potential bacteria against the Vibrio spp. population growth in TCBSA media. This study highlights the potential of whiteleg shrimp digestive symbiont bacteria to enhance immunity against vibriosis while providing safe digestive enzymatic potential for whiteleg shrimp and humans as a potential probiotic candidate.
References
Afrin, S. et al., 2024. Characterization of protease-producing bacteria from garden soil and antagonistic activity against pathogenic bacteria. The Microbe, 4, 100123. doi: 10.1016/j.microb.2024.100123
Alloul, A. et al., 2021. Purple bacteria as added-value protein ingredient in shrimp feed: Penaeus vannamei growth performance, and tolerance against Vibrio and ammonia stress. Aquaculture, 530(August 2020), 735788. doi: 10.1016/j.aquaculture.2020.735788
Amatul-Samahah, M.A. et al., 2020. Vaccination trials against vibriosis in shrimp: A review. Aquaculture Reports, 18, 100471. doi: 10.1016/j.aqrep.2020.100471
Amelia, F. et al., 2020. Diversity Analysis of Genus Vibrio in Enterocytozoon Hepatopenaei (EHP) Infected Shrimp with Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC PCR) Method. Journal of Aquaculture and Fish Health, 9(3), pp.238-251. doi: 10.20473/jafh.v9i3.18855
Amiin, M.K. et al., 2023. The role of probiotics in vannamei shrimp aquaculture performance - A review. Veterinary World, 16(3), pp.638–649. doi: 10.14202/vetworld.2023.638-649
Chen, L. et al., 2021. Effects of Bacillus velezensis Supplementation on the Growth Performance, Immune Responses, and Intestine Microbiota of Litopenaeus vannamei. Frontiers in Marine Science, 8, 744281. doi: 10.3389/fmars.2021.744281
Dayalan, V. et al., 2022. Studies on temperature impact (sudden and gradual) of the white-leg shrimp Litopenaeus vannamei. Environmental Science and Pollution Research, 31(27), pp.38743–38750. doi: 10.1007/s11356-022-20963-y
Devi, A. R., Susilowati, A., & Setyaningsih, R. 2019. Morphology, molecular identification, and pathogenicity of Vibrio spp. on blood clam (Anadara granosa) in Yogyakarta, Indonesia tourism beach areas. Biodiversitas Journal of Biological Diversity, 20(10), pp.2890-2896. doi: 10.13057/biodiv/d201016
Duan, Y. et al., 2020. Response of intestine microbiota, digestion, and immunity in Pacific white shrimp Litopenaeus vannamei to dietary succinate. Aquaculture, 517, 734762. doi: 10.1016/j.aquaculture.2019.734762
EL-Sayed, A. I. M. et al., 2022. Identification of Lactobacillus strains from human mother milk and cottage cheese revealed potential probiotic properties with enzymatic activity. Scientific Reports, 12(1), 22522. doi: 10.1038/s41598-022-27003-2
Fernando, O., Susena, K. C., & Herlin. 2021. Efficiency Analysis of Shrimp Cultivation in Kaur District (Case Study Of South Kaur District) Analisis Efisiensi Budidaya Tambak Udang Di Kabupaten Kaur (Studi Kasus Kecamatan Kaur Selatan). Jurnal Ekonomi Manajemen Akuntansi Dan Kuangan, 2(1), pp.39–54.
Hembrom, P. S. et al., 2023. Influence of gut microbiome on health and development of penaeid shrimps. Aquatic Sciences, 86, 4. doi: 10.1007/s00027-023-01018-x
Joshi, V. B. et al., 2022. Extraction, Purification And Characterization Of Amylase, Protease, Lipase Of Halophilic Bacterial Isolates From Bordi, India. International Journal of Applied Biology, 6(1), pp.63–72. doi: 10.20956/ijab.v6i1.19118
Kristiany, M.G.E. et al., 2022. Effectiveness of Different Probiotic Application on Vannamei Shrimp Productivity. Jurnal Perikanan Universitas Gadjah Mada, 24(1), pp.31-35. doi: 10.22146/jfs.68672
Lay, B.W., 1994. Analisis Mikroba di Laboratorium, Jakarta: PT Rajagrafindo Persada.
Leber, A.L., 2016. Clinical Microbiology Procedures Handbook, ASM Press. doi: 10.1128/9781555818814
Mohamad, N. et al., 2020. Screening of Lactic Acid Bacteria isolated from giant freshwater prawn (Macrobrachium rosenbergii) as potential probiotics. Aquaculture Reports, 18, 100523. doi: 10.1016/j.aqrep.2020.100523
Mufidah, D.A. & Diantari, R., 2019. Penggunaan Ekstrak Buah Mangrove (Avicennia sp.) Sebagai Bahan Antibakteri Alami Untuk Pengobatan Penyakit Yang Disebabkan Oleh Vibrio parahaemolyticus Pada Udang Vaname (Litopenaeus vannamei). AQUASAINS (Jurnal Ilmu Perikanan Dan Sumberdaya Perairan) Penggunaan, 1, 5.
Mustafa, A. et al., 2023. Strategy for Developing Whiteleg Shrimp (Litopenaeus vannamei) Culture Using Intensive/Super-Intensive Technology in Indonesia. Sustainability (Switzerland), 15(3), 1753. doi: 10.3390/su15031753
Priyanto, J.A. et al., 2023. The Antibacterial and Antibiofilm Activities of the Endophytic Bacteria Associated with Archidendron pauciflorum against Multidrug ‑ Resistant Strains. Applied Biochemistry and Biotechnology, 195(11), pp.6653-6674. doi: 10.1007/s12010-023-04382-4
Proespraiwong, P. et al., 2023. Evaluation of Bacillus spp. as Potent Probiotics with Reduction in AHPND-Related Mortality and Facilitating Growth Performance of Pacific White Shrimp (Litopenaeus vannamei) Farms. Microorganisms, 11(9), 2176. doi: 10.3390/microorganisms11092176
Rusmana, I. et al., 2021a. Characteristics and Activity Anti Qourum Sensing Bacillus spp. Isolated from Penaeus vannamei Shrimp Ponds. HAYATI Journal of Biosciences, 29(1), pp.97–106. doi: 10.4308/hjb.29.1.97-106
Rusmana, I., Isramilda, I., & Akhdiya, A., 2021b. Characteristics of anti-Vibrio harveyi compounds produced by Bacillus spp. isolated from shrimp ponds. Biodiversitas Journal of Biological Diversity, 22(11), pp.4872-4879. doi: 10.13057/biodiv/d221120
Situbondo, B. 2021. Budidaya Udang Vaname (Litopenaeus vannamei) di Tambak Millenial Shrimp Farming (MSF) Bpbap Takalar, Sulawesi Selatan. UNIVERSITAS NUSA NIPA.
Taheri-Araghi, S., 2024. Synergistic action of antimicrobial peptides and antibiotics: current understanding and future directions. Frontiers in Microbiology, 15, 1390765. doi: 10.3389/fmicb.2024.1390765
Torpee, S. et al., 2021. Dietary supplementation with probiotic Rhodobacter sphaeroides SS15 extract to control acute hepatopancreatic necrosis disease (AHPND)-causing Vibrio parahaemolyticus in cultivated white shrimp. Journal of Invertebrate Pathology, 186, 107585. doi: 10.1016/j.jip.2021.107585
