INVESTIGATION OF THE IN VITRO ANTIMICROBIAL AND ANTI-BIOFILM ACTIVITIES OF STREPTOMYCES SP. SS532 AGAINST AEROMONAS HYDROPHILA CAUSING SEPTICEMIA IN PANGASIUS
Main Article Content
Abstract
Aeromonas hydrophila causes fish septicemia and substantial economic losses in Vietnamese pangasius farming, creating a need for effective control of this pathogen in aquaculture. To address this need, this study isolated a bacterial strain from a water sample collected in Dong Thap Province, Vietnam, and found that it exhibited antibacterial activity against A. hydrophila. The isolate was identified as Streptomyces sp. SS532 based on morphological characteristics and the 16S rRNA gene. Its antibacterial activity remained stable at high temperatures and across a wide pH range and was retained after exposure to ultraviolet radiation and protease treatment. SS532 also exhibited relatively broad-spectrum activity against other bacterial pathogens, which was maintained over nine successive generations. The ethyl acetate extract from the SS532 culture had a minimum inhibitory concentration (MIC) of 640 mg/mL against A. hydrophila and showed strong antibiofilm activity at concentrations from 0.04× MIC. These findings provide a basis for further research on the use of Streptomyces sp. SS532 to prevent and control A. hydrophila-induced septicemia in fish.
Keywords
Aeromonas hydrophila, antibacterial activity, pangasius, Streptomyces sp. SS532, Vietnam
Article Details
References
Alam, K., Mazumder, A., Sikdar, S., Zhao, Y.-M., Hao, J., Song, C., Wang, Y., Sarkar, R., Islam, S., Zhang, Y., & Li, A. (2022). Streptomyces: The biofactory of secondary metabolites. Frontiers in Microbiology, 13, 968053. https://doi.org/10.3389/fmicb.2022.968053
Balouiri, M., Sadiki, M., & Ibnsouda, S. K. (2016). Methods for in vitro evaluating antimi-crobial activity: A review. Journal of Pharmaceutical Analysis, 6(2), 71-79. https://doi.org/10.1016/j.jpha.2015.11.005
Bui, T. V. H. (2006). Nghiên cứu xạ khuẩn sinh chất kháng sinh chống nấm gây bệnh thực vật ở Việt Nam [Research on actinomycetes producing antifungal antibiotic substances against plant pathogenic fungi in Vietnam] [Doctoral dissertation in Biology, VNU Uni-versity of Science, Vietnam National University, Hanoi]. VNU University of Science, Vietnam National University, Hanoi.
Campanile, F., Wootton, M., Davies, L., Aprile, A., Mirabile, A., Pomponio, S., Demetrio, F., Bongiorno, D., Walsh, T. R., Stefani, S., & Mezzatesta, M. L. (2020). Gold standard susceptibility testing of fosfomycin in Staphylococcus aureus and Enterobacterales us-ing a new agar dilution panel®. Journal of Global Antimicrobial Resistance, 23, 334–337. https://doi.org/10.1016/j.jgar.2020.08.025
Dang, T. H. O., Xuan, T. T. T., Duyen, L. T. M., Le, N. P., & Hoang, H. A. (2021). Protective efficacy of phage PVN02 against haemorrhagic septicaemia in striped catfish Pangasianodon hypophthalmus via oral administration. Journal of Fish Diseases, 44(8), 1255–1263. https://doi.org/10.1111/jfd.13387
Derome, N., Gauthier, J., Boutin, S., & Llewellyn, M. (2016). Bacterial Opportunistic Pathogens of Fish. In: Hurst, C. (eds) The Rasputin Effect: When Commensals and Symbionts Become Parasitic. Advances in Environmental Microbiology, 3. https://doi.org/10.1007/978-3-319-28170-4
Downes, K. J., & Goldman, J. L. (2021). Too Much of a Good Thing: Defining Antimicrobial Therapeutic Targets to Minimize Toxicity. Clinical Pharmacology & Therapeutics, 109(4), 905–917. https://doi.org/10.1002/cpt.2190
Falagas, M. E., Tansarli, G. S., Rafailidis, P. I., Kapaskelis, A., & Vardakas, K. Z. (2012). Impact of Antibiotic MIC on Infection Outcome in Patients with Susceptible Gram-Negative Bacteria: A Systematic Review and Meta-Analysis. Antimicrobial Agents and Chemotherapy, 56(8), 4214–4222. https://doi.org/10.1128/AAC.00663-12
Hackmann, T. J. (2025). Setting new boundaries of 16S rRNA gene identity for prokaryotic taxonomy. International Journal of Systematic and Evolutionary, 75(006747). https://doi.org/10.1099/ijsem.0.006747
Huynh, D. A., Trinh, T. A., Pham, N. G. H., Tran, T. N., Le, T. T., & Nguyen, H. C. (2024). Phân lập và nghiên cứu hoạt tính kháng Staphylococcus aureus kháng methicillin của các chủng xạ khuẩn phân lập từ tỉnh Gia Lai, Việt Nam [Isolation and antibacterial ac-tivity study against methicillin-resistant Staphylococcus aureus of actinomycete strains isolated from Gia Lai province, Vietnam]. Journal of Military Pharmaco-medicine, 49(3), 76–86. https://doi.org/10.56535/jmpm.v49i3.564
Jeamsripong, S., Odoi, J. O., Shahi, M. K., Anuntawirun, S., Roongrojmongkhon, N., & Thiptara, A. (2025). Global spread and antimicrobial resistance of Aeromonas hydrophila in aquatic food animals: A systematic review and meta-analysis. Scientific Reports, 15(1), 28441. https://doi.org/10.1038/s41598-025-14498-8
Kieser, T., Bibb, M. J., Buttner, M. J., Chater, K. F., & Hopwood, D. A. (2000). Practical Streptomyces genetics: A laboratory manual. The John Innes Foundation.
Landwehr, W., Kämpfer, P., Glaeser, S. P., Rückert, C., Kalinowski, J., Blom, J., Goesmann, A., Mack, M., Schumann, P., Atasayar, E., Hahnke, R. L., Rohde, M., Martin, K., Stadler, M., & Wink, J. (2018). Taxonomic analyses of members of the Streptomyces cinnabarinus cluster, description of Streptomyces cinnabarigriseus sp. nov. and Streptomyces davaonensis sp. nov. International Journal of Systematic and Evolution-ary, 68(1), 382-393. https://doi.org/10.1099/ijsem.0.002519
Lei, K.-H., & Lai, H.-T. (2019). Effects of sunlight, microbial activity, and temperature on the declines of antibiotic lincomycin in freshwater and saline aquaculture pond waters and sediments. Environmental Science and Pollution Research, 26(33), 33988–33994. https://doi.org/10.1007/s11356-018-3006-y
Ljubojević Pelić, D., Radosavljević, V., Pelić, M., Živkov Baloš, M., Puvača, N., Jug-Dujaković, J., & Gavrilović, A. (2024). Antibiotic Residues in Cultured Fish: Implications for Food Safety and Regulatory Concerns. Fishes, 9(12), 484. https://doi.org/10.3390/fishes9120484
Long, W., Zhao, W., He, L., Khan, T. A., Lai, X., Sun, Y., Huang, W., Yi, G., & Xia, L. (2024) Streptomyces enissocaesilis L-82 has broad-spectrum antibacterial activity and pro-motes growth for Carassius auratus. Applied Microbiology and Biotechnology, 108(1):220. https://doi.org/10.1007/s00253-024-13031-7
Manandhar, S., Singh, A., Varma, A., Pandey, S., & Shrivastava, N. (2018). Evaluation of methods to detect in vitro biofilm formation by staphylococcal clinical isolates. BMC Research Notes, 11, 714. https://doi.org/10.1186/s13104-018-3820-9
Mispelaere, M., De Rop, A.-S., Hermans, C., De Maeseneire, S. L., Soetaert, W. K., De Mol, M. L., & Hulpiau, P. (2024). Whole genome–based comparative analysis of the genus Streptomyces reveals many misclassifications. Applied Microbiology and Biotechnolo-gy, 108, 453. https://doi.org/10.1007/s00253-024-13290-4
Montanari, E., Bernardo, G., Le Noci, V., Anselmi, M., Pupa, S. M., Tagliabue, E., Sommariva, M., & Sfondrini, L. (2025). Biofilm formation by the host microbiota: A protective shield against immunity and its implication in cancer. Molecular Cancer, 24(1), 148. https://doi.org/10.1186/s12943-025-02348-0
Phan, L. T., Bui, T. M., Nguyen, T. T. T., Gooley, G. J., Ingram, B. A., Nguyen, H. V., Nguyen, P. T., & De Silva, S. S. (2009). Current status of farming practices of striped catfish, Pangasianodon hypophthalmus in the Mekong Delta, Vietnam. Aquaculture, 296(3), 227–236. https://doi.org/10.1016/j.aquaculture.2009.08.017
Quach, V. C. T., Tran, Q. D., Ha, N. H., Nguyen, B. T., Tu, T. D., & Nguyen, P. T. (2024). Antibacterial activity of Streptomyces spp. against Aeromonas hydrophila causing hemorrhagic disease in intensively cultured pangasius catfish. Veterinary Integrative Sciences, 22(1), 73–91. https://doi.org/10.12982/VIS.2024.007
Richard, U. O., Chukwura, E. I., & Mercy, I. E. (2020). Physicochemical, Bacteriological and Parasitological Examination of Selected Fish Pond Water Samples in Awka and Its Environment Anambra State, Nigeria. Journal of Advances in Microbiology, 27–48. https://doi.org/10.9734/jamb/2020/v20i330226
Quach, N. T., Vu, T.-H.-N., Nguyen, T. T. A., Bui, T. L., Le, T. T. X., & Phi, Q. T. (2022). Ex-ploring antibacterial and cytotoxic potential of endophytic Streptomyces isolated from the medicinal plant Litsea cubeba (Lour.) Pers. Vietnam Journal of Science and Tech-nology, 60(5), 756–766. https://doi.org/10.15625/2525-2518/16669
Sarker, J., & Faruk, M. a. R. (2016). Experimental infection of Aeromonas hydrophila in pangasius. Progressive Agriculture, 27(3), 392–399. https://doi.org/10.3329/pa.v27i3.30836
Schloss, P. D., & Handelsman, J. (2005). Introducing DOTUR, a Computer Program for Defining Operational Taxonomic Units and Estimating Species Richness. Applied and Environmental Microbiology, 71(3), 1501–1506. https://doi.org/10.1128/AEM.71.3.1501-1506.2005
Selvakumar, D., Arun, K., Suguna, S., Kumar, D., & Dhevendaran, K. (2010). Bioactive potential of Streptomyces against fish and shellfish pathogens. Iranian Journal of Microbiology, 2(3), 157–164. PMID:22347566. https://pubmed.ncbi.nlm.nih.gov/22347566/
Taguri, T., Tanaka, T., & Kouno, I. (2006). Antibacterial Spectrum of Plant Polyphenols and Extracts Depending upon Hydroxyphenyl Structure. Biological and Pharmaceutical Bulletin, 29(11), 2226–2235. https://doi.org/10.1248/bpb.29.2226
Zheng, S., Wang, Y., Chen, C., Zhou, X., Liu, Y., Yang, J., Geng, Q., Chen, G., Ding, Y., & Yang, F. (2022). Current Progress in Natural Degradation and Enhanced Removal Techniques of Antibiotics in the Environment: A Review. International Journal of Environmental Research and Public Health, 19(17), 10919. https://doi.org/10.3390/ijerph191710919
Balouiri, M., Sadiki, M., & Ibnsouda, S. K. (2016). Methods for in vitro evaluating antimi-crobial activity: A review. Journal of Pharmaceutical Analysis, 6(2), 71-79. https://doi.org/10.1016/j.jpha.2015.11.005
Bui, T. V. H. (2006). Nghiên cứu xạ khuẩn sinh chất kháng sinh chống nấm gây bệnh thực vật ở Việt Nam [Research on actinomycetes producing antifungal antibiotic substances against plant pathogenic fungi in Vietnam] [Doctoral dissertation in Biology, VNU Uni-versity of Science, Vietnam National University, Hanoi]. VNU University of Science, Vietnam National University, Hanoi.
Campanile, F., Wootton, M., Davies, L., Aprile, A., Mirabile, A., Pomponio, S., Demetrio, F., Bongiorno, D., Walsh, T. R., Stefani, S., & Mezzatesta, M. L. (2020). Gold standard susceptibility testing of fosfomycin in Staphylococcus aureus and Enterobacterales us-ing a new agar dilution panel®. Journal of Global Antimicrobial Resistance, 23, 334–337. https://doi.org/10.1016/j.jgar.2020.08.025
Dang, T. H. O., Xuan, T. T. T., Duyen, L. T. M., Le, N. P., & Hoang, H. A. (2021). Protective efficacy of phage PVN02 against haemorrhagic septicaemia in striped catfish Pangasianodon hypophthalmus via oral administration. Journal of Fish Diseases, 44(8), 1255–1263. https://doi.org/10.1111/jfd.13387
Derome, N., Gauthier, J., Boutin, S., & Llewellyn, M. (2016). Bacterial Opportunistic Pathogens of Fish. In: Hurst, C. (eds) The Rasputin Effect: When Commensals and Symbionts Become Parasitic. Advances in Environmental Microbiology, 3. https://doi.org/10.1007/978-3-319-28170-4
Downes, K. J., & Goldman, J. L. (2021). Too Much of a Good Thing: Defining Antimicrobial Therapeutic Targets to Minimize Toxicity. Clinical Pharmacology & Therapeutics, 109(4), 905–917. https://doi.org/10.1002/cpt.2190
Falagas, M. E., Tansarli, G. S., Rafailidis, P. I., Kapaskelis, A., & Vardakas, K. Z. (2012). Impact of Antibiotic MIC on Infection Outcome in Patients with Susceptible Gram-Negative Bacteria: A Systematic Review and Meta-Analysis. Antimicrobial Agents and Chemotherapy, 56(8), 4214–4222. https://doi.org/10.1128/AAC.00663-12
Hackmann, T. J. (2025). Setting new boundaries of 16S rRNA gene identity for prokaryotic taxonomy. International Journal of Systematic and Evolutionary, 75(006747). https://doi.org/10.1099/ijsem.0.006747
Huynh, D. A., Trinh, T. A., Pham, N. G. H., Tran, T. N., Le, T. T., & Nguyen, H. C. (2024). Phân lập và nghiên cứu hoạt tính kháng Staphylococcus aureus kháng methicillin của các chủng xạ khuẩn phân lập từ tỉnh Gia Lai, Việt Nam [Isolation and antibacterial ac-tivity study against methicillin-resistant Staphylococcus aureus of actinomycete strains isolated from Gia Lai province, Vietnam]. Journal of Military Pharmaco-medicine, 49(3), 76–86. https://doi.org/10.56535/jmpm.v49i3.564
Jeamsripong, S., Odoi, J. O., Shahi, M. K., Anuntawirun, S., Roongrojmongkhon, N., & Thiptara, A. (2025). Global spread and antimicrobial resistance of Aeromonas hydrophila in aquatic food animals: A systematic review and meta-analysis. Scientific Reports, 15(1), 28441. https://doi.org/10.1038/s41598-025-14498-8
Kieser, T., Bibb, M. J., Buttner, M. J., Chater, K. F., & Hopwood, D. A. (2000). Practical Streptomyces genetics: A laboratory manual. The John Innes Foundation.
Landwehr, W., Kämpfer, P., Glaeser, S. P., Rückert, C., Kalinowski, J., Blom, J., Goesmann, A., Mack, M., Schumann, P., Atasayar, E., Hahnke, R. L., Rohde, M., Martin, K., Stadler, M., & Wink, J. (2018). Taxonomic analyses of members of the Streptomyces cinnabarinus cluster, description of Streptomyces cinnabarigriseus sp. nov. and Streptomyces davaonensis sp. nov. International Journal of Systematic and Evolution-ary, 68(1), 382-393. https://doi.org/10.1099/ijsem.0.002519
Lei, K.-H., & Lai, H.-T. (2019). Effects of sunlight, microbial activity, and temperature on the declines of antibiotic lincomycin in freshwater and saline aquaculture pond waters and sediments. Environmental Science and Pollution Research, 26(33), 33988–33994. https://doi.org/10.1007/s11356-018-3006-y
Ljubojević Pelić, D., Radosavljević, V., Pelić, M., Živkov Baloš, M., Puvača, N., Jug-Dujaković, J., & Gavrilović, A. (2024). Antibiotic Residues in Cultured Fish: Implications for Food Safety and Regulatory Concerns. Fishes, 9(12), 484. https://doi.org/10.3390/fishes9120484
Long, W., Zhao, W., He, L., Khan, T. A., Lai, X., Sun, Y., Huang, W., Yi, G., & Xia, L. (2024) Streptomyces enissocaesilis L-82 has broad-spectrum antibacterial activity and pro-motes growth for Carassius auratus. Applied Microbiology and Biotechnology, 108(1):220. https://doi.org/10.1007/s00253-024-13031-7
Manandhar, S., Singh, A., Varma, A., Pandey, S., & Shrivastava, N. (2018). Evaluation of methods to detect in vitro biofilm formation by staphylococcal clinical isolates. BMC Research Notes, 11, 714. https://doi.org/10.1186/s13104-018-3820-9
Mispelaere, M., De Rop, A.-S., Hermans, C., De Maeseneire, S. L., Soetaert, W. K., De Mol, M. L., & Hulpiau, P. (2024). Whole genome–based comparative analysis of the genus Streptomyces reveals many misclassifications. Applied Microbiology and Biotechnolo-gy, 108, 453. https://doi.org/10.1007/s00253-024-13290-4
Montanari, E., Bernardo, G., Le Noci, V., Anselmi, M., Pupa, S. M., Tagliabue, E., Sommariva, M., & Sfondrini, L. (2025). Biofilm formation by the host microbiota: A protective shield against immunity and its implication in cancer. Molecular Cancer, 24(1), 148. https://doi.org/10.1186/s12943-025-02348-0
Phan, L. T., Bui, T. M., Nguyen, T. T. T., Gooley, G. J., Ingram, B. A., Nguyen, H. V., Nguyen, P. T., & De Silva, S. S. (2009). Current status of farming practices of striped catfish, Pangasianodon hypophthalmus in the Mekong Delta, Vietnam. Aquaculture, 296(3), 227–236. https://doi.org/10.1016/j.aquaculture.2009.08.017
Quach, V. C. T., Tran, Q. D., Ha, N. H., Nguyen, B. T., Tu, T. D., & Nguyen, P. T. (2024). Antibacterial activity of Streptomyces spp. against Aeromonas hydrophila causing hemorrhagic disease in intensively cultured pangasius catfish. Veterinary Integrative Sciences, 22(1), 73–91. https://doi.org/10.12982/VIS.2024.007
Richard, U. O., Chukwura, E. I., & Mercy, I. E. (2020). Physicochemical, Bacteriological and Parasitological Examination of Selected Fish Pond Water Samples in Awka and Its Environment Anambra State, Nigeria. Journal of Advances in Microbiology, 27–48. https://doi.org/10.9734/jamb/2020/v20i330226
Quach, N. T., Vu, T.-H.-N., Nguyen, T. T. A., Bui, T. L., Le, T. T. X., & Phi, Q. T. (2022). Ex-ploring antibacterial and cytotoxic potential of endophytic Streptomyces isolated from the medicinal plant Litsea cubeba (Lour.) Pers. Vietnam Journal of Science and Tech-nology, 60(5), 756–766. https://doi.org/10.15625/2525-2518/16669
Sarker, J., & Faruk, M. a. R. (2016). Experimental infection of Aeromonas hydrophila in pangasius. Progressive Agriculture, 27(3), 392–399. https://doi.org/10.3329/pa.v27i3.30836
Schloss, P. D., & Handelsman, J. (2005). Introducing DOTUR, a Computer Program for Defining Operational Taxonomic Units and Estimating Species Richness. Applied and Environmental Microbiology, 71(3), 1501–1506. https://doi.org/10.1128/AEM.71.3.1501-1506.2005
Selvakumar, D., Arun, K., Suguna, S., Kumar, D., & Dhevendaran, K. (2010). Bioactive potential of Streptomyces against fish and shellfish pathogens. Iranian Journal of Microbiology, 2(3), 157–164. PMID:22347566. https://pubmed.ncbi.nlm.nih.gov/22347566/
Taguri, T., Tanaka, T., & Kouno, I. (2006). Antibacterial Spectrum of Plant Polyphenols and Extracts Depending upon Hydroxyphenyl Structure. Biological and Pharmaceutical Bulletin, 29(11), 2226–2235. https://doi.org/10.1248/bpb.29.2226
Zheng, S., Wang, Y., Chen, C., Zhou, X., Liu, Y., Yang, J., Geng, Q., Chen, G., Ding, Y., & Yang, F. (2022). Current Progress in Natural Degradation and Enhanced Removal Techniques of Antibiotics in the Environment: A Review. International Journal of Environmental Research and Public Health, 19(17), 10919. https://doi.org/10.3390/ijerph191710919