Gut bacteria isolates of Hirudinaria manillensis and its antibacterial activities
DOI:
https://doi.org/10.24191/bioenv.v4i2.139Keywords:
Leeches , Southeast Asian freshwater , sanquivorousAbstract
Leeches have been used in traditional and modern medical practices for centuries; however, information on leech species identification, gut-associated bacterial communities, and their potential antimicrobial interactions remains limited. This study aimed to identify leech specimens and isolate and characterise their gut-associated microbiota with potential antibacterial activity. Leech samples were collected from two locations: Kemaman, Terengganu, and Jengka, Pahang, Malaysia. Species identification was conducted based on external and internal morphological characteristics. Gut bacteria were isolated and differentiated according to colony morphology and cellular characteristics. The antibacterial activity of the isolates was evaluated using cross-streaking and disk diffusion assays against selected pathogenic bacteria, namely Bacillus subtilis, Staphylococcus aureus, and Escherichia coli. Morphological analysis confirmed that all leech specimens belonged to the dark buffalo leech, Hirudinaria manillensis. A total of three bacterial isolates were successfully obtained from the leech gut, of which two showed observable zones of inhibition against the tested bacteria. Overall, this study provides baseline information on the morphology of H. manillensis and its associated gut microbiota. The findings enhance current understanding of leech–microbe interactions and provide a basis for future studies on the antimicrobial potential of leech-associated bacteria.
References
Alberdi, A., Martin Bideguren, G., & Aizpurua, O. (2021). Diversity and compositional changes in the gut microbiota of wild and captive vertebrates: a meta-analysis. Scientific Reports, 11(1), 22660. https://doi.org/10.1038/s41598-021-02015-6
Arias, A., Surugiu, V., Carballeira, R., Popa, O. P., Popa, L. O., & Utevsky, S. (2021). Unravelling the extent of diversity within the Iberian medicinal leeches (Hirudinea: Hirudo) using molecules and morphology. Biology, 10(4), 315. https://doi.org/10.3390/biology10040315
Bam, J., Islam, S., & Pathak, P. (2025). Ultrastructural jaw morphology of Hirudinaria manillensis and Haemadipsa montana. International Journal of Advanced Biochemistry Research 9(6), 81-85. https://www.doi.org/10.33545/26174693.2025.v9.i6Sb.4495
Chong, L. K., Ong, A. H., Tan, S. G., Taranjeet, K. A. S., Peris, M. M., Sana, A. M. M. A., & Hassan, H. R. (2014). Morphological and genetic variations of the freshwater leech, Hirudinaria spp., in Peninsular Malaysia. Biochemical Genetics, 52(5), 283-295. https://doi.org/10.1007/s10528-014-9647-8
Chuang, C., Qin Ren, Q., & Farra Aidah Jumuddin, F. A. (2022). Low-Cost Purification Cultivation Process For Medical "Hirudinaria Manillensis". Journal of Advanced Zoology, 41, 461-465. https://doi.org/10.53555/jaz.v43iS1.4057
Davido, B., Merrick, B., Kuijper, E., Benech, N., Biehl, L. M., & Corcione, S. (2025). How can the gut microbiome be targeted to fight multidrug-resistant organisms? The Lancet Microbe, 6(8), 101063. https://doi.org/10.1016/j.lanmic.2024.101063
de Jonge, N., Carlsen, B., Christensen, M. H., Pertoldi, C., & Nielsen, J. L. (2022). The gut microbiome of 54 mammalian species. Frontiers in Microbiology, 13, 886252. https://doi.org/10.3389/fmicb.2022.886252
Dilmore, L. A., & Hood, M. A. (1986). Vibrios of some deep-water invertebrates. FEMS Microbiology Letters, 35(2-3), 221-224. https://doi.org/10.1111/j.1574-6968.1986.tb01531.x
Guan, D. L., Yang, J., Liu, Y. K., Li, Y., Mi, D., Ma, L. B., Wang, Z. Z., Xu, S. Q., & Qiu, Q. (2020). Draft genome of the Asian buffalo leech Hirudinaria manillensis. Frontiers in Genetics, 10, 1321. https://doi.org/10.3389/fgene.2019.01321
Hosseini, M., Jadidi, A., Derakhshan Barjoei, M. M., & Salehi, M. (2024). Applications of leech therapy in medicine: a systematic review. Frontiers in Medicine, 11, 1417041. https://doi.org/10.3389/fmed.2024.1417041
Jeratthitikul, E., Jiranuntskul, P., Nakano, T., Sutcharit, C., & Panha, S. (2020). A new species of buffalo leech in the genus Hirudinaria Whitman, 1886 (Arhynchobdellida, Hirudinidae) from Thailand. ZooKeys, 933, 1-14. https://doi.org/10.3897/zookeys.933.49314
Kamada, N., Seo, S. U., Chen, G. Y., & Núñez, G. (2013). Role of the gut microbiota in immunity and inflammatory disease. Nature Reviews Immunology, 13(5), 321-335. https://doi.org/10.1038/nri3430
Karasartova, D., Arslan-Akveran, G., Sensoz, S., Mumcuoglu, K. Y., & Taylan-Ozkan, A. (2025). Hirudo verbana Microbiota Dynamics: A Key Factor in Hirudotherapy-Related Infections?. Microorganisms, 13(4), 918. https://doi.org/10.3390/microorganisms13040918
Kaur, A., Guerrero‐Garzón, J. F., Rasheed, S., Zehl, M., Fries, F., Morgenstern, B., Zotchev, S. B., & Müller, R. (2025). Saarvienin A—A novel glycopeptide with potent activity against Drug‐Resistant bacteria. Angewandte Chemie, 137(25), e202425588. https://doi.org/anie.202425588
Olsen, R. E., Sundell, K., Hansen, T., Hemre, G. I., Myklebust, R., Mayhew, T. M., & Ringø, E. (2002). Acute stress alters the intestinal lining of Atlantic salmon, Salmo salar L.: An electron microscopical study. Fish Physiology and Biochemistry, 26(3), 211-221. https://doi.org/10.1023/A:1026217719534
Petersen, J.M., & Osvatic. J. (2018). Microbiomes in natura: Importance of invertebrates in understanding the natural variety of animal-microbe interactions. mSystems, 3(2), 10-1128. https://doi.org/10.1128/msystems.00179-17
Phillips, A. J., Govedich, F. R., & Moser, W. E. (2020). Leeches in the extreme: Morphological, physiological, and behavioral adaptations to inhospitable habitats. International Journal for Parasitology: Parasites and Wildlife, 12, 318–325. https://doi.org/10.1016/j.ijppaw.2020.09.003
Sidhu, P. K., & Nehra, K. (2021). Bacteriocins of Lactic acid bacteria as potent antimicrobial peptides against food pathogens. In: Habib, M. K., & Martín-Gómez, C. (Eds) Biomimetics. IntechOpen, London. https://doi.org/10.5772/intechopen.95747
Silver L. L. (2025). Lasso-shaped molecule is a new type of broad-spectrum antibiotic. Nature, 640(8060), 887–889. https://doi.org/10.1038/d41586-025-00901-x
Solis-Balandra, M. A., & Sanchez-Salas, J. L. (2024). Classification and multi-functional use of bacteriocins in health, biotechnology, and food industry. Antibiotics, 13(7), 666. https://doi.org/10.3390/antibiotics13070666
Suman, V., & Yugandhar, N. M. (2021). Characterization of pectinase and antibacterial activity by using Nocardiopsis dasnonivelli S10 isolated from marine samples. Journal of Cardiovascular Disease Research, 12(1), 616-631. https://doi.org/10.48047/
Sun, Y., Shi, X., Xing, Y., Ren, X. X., Zhang, D. Y., Li, X., Xiu, Z. L., & Dong, Y. S. (2022). Co-culture of Aspergillus sydowii and Bacillus subtilis induces the production of antibacterial metabolites. Fungal biology, 126(4), 320–332. https://doi.org/10.1016/j.funbio.2022.01.002
Tubtimon, J., Jeratthitikul, E., Sutcharit, C., Kongim, B., & Panha, S. (2014). Systematics of the freshwater leech genus Hirudinaria Whitman, 1886 (Arhynchobdellida, Hirudinidae) from Northeastern Thailand. ZooKeys, 452, 15–33. https://doi.org/10.3897/zookeys.452.7528
Ullah, H., Arbab, S., Tian, Y., Chen, Y., Liu, C. Q., Li, Q., & Li, K. (2024). Crosstalk between gut microbiota and host immune system and its response to traumatic injury. Frontiers in Immunology, 15, 1413485. https://doi.org/10.3389/fimmu.2024.1413485
Xie, Y., Xu, S., Xi, Y., Li, Z., Zuo, E., Xing, K., Bai, L., & Li, K. (2024), Global meta-analysis reveals the drivers of gut microbiome variation across vertebrates. IMetaOmics, 1, e35. https://doi.org/10.1002/imo2.35
Xue, J., Ajuwon, K. M., & Fang, R. (2020). Mechanistic insight into the gut microbiome and its interaction with host immunity and inflammation. Animal Nutrition, 6(4), 421-428. https://doi.org/10.1016/j.aninu.2020.05.007
Yun, J. H., Roh, S. W., Whon, T. W., Jung, M. J., Kim, M. S., Park, D. S., Yoon, C., Nam, Y. D., Kim, Y. J., Choi, J. H., Kim, J. Y., Shin, N. R., Kim, S. H., Lee, W. J., & Bae, J. W. (2014). Insect gut bacterial diversity determined by environmental habitat, diet, developmental stage, and phylogeny of host. Applied and Environmental Microbiology, 80(17), 5254–5264. https://doi.org/10.1128/AEM.01226-14
© 2026 by the authors. Submitted for possible open access publication under the terms and conditions of the Creative Commons Attribution (CC BY-NC-ND) license (http://creativecommons.org/licenses/by/4.0/).
Zulhisyam, A. K., Jamaludin, M.H., Shazani, S., Amizi, M., Lee, S. W., Sukhairi, M.R.M., Mazlan, M., Iqbal, A. M., Anwar Ismail, A., Azwanida, N. N., Marwan, M., & Farhan, A. M. (2014). A study on the anatomy features of green and dark brown buffalo leech (Hirudinea manillensis). Journal of Sustainability Science and Management, 9, 134–140.
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