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Penapisan senyawa bioaktif antibakteri fungi laut endofit asal Pulau Buton Sulawesi Tenggara Screening bioactive compounds of marine endophytic fungi as antibacteria from Buton Island, Southeast Sulawesi
Abstract
Marine endophytic fungi harbor inherent bioactive chemicals that exhibit antimicrobial, antitumor, antiviral, anticancer, and anti-inflammatory properties. The aim of this study was to assess the capability of bioactive chemicals found in endophytic marine fungi to prevent the growth of the harmful bacterium Vibrio harveyi. This research was carried out using a series of procedures, including isolation, selection, antagonistic testing, extraction of active compounds, and antibacterial testing. The process of isolating and selecting endophytic marine fungi was conducted on seaweed and mangrove samples obtained from Buton Island, Southeast Sulawesi. This was performed using Potato Dextrose Agar (PDA). 35 isolates were subjected to antagonistic testing against the pathogenic bacterium V. harveyi, which led to the formation of clear zones with sizes ranging from 2 mm to 21 mm. Extraction was conducted using a maceration technique with ethyl acetate as the solvent. The resulting extract was then concentrated using a rotary evaporator. The extract derived from the marine fungus Aspergillus terreus (WB 1-2) showed the largest inhibitory zone, measuring 21.8±0.3 mm, when tested at a dosage of 2 mg/mL. The diameter of the inhibitory zone of the A. terreus extract was greater than that of the positive control, chloramphenicol (19 mm). These findings suggest that the marine endophytic fungus A. terreus can be used as a natural antibacterial agent to inhibit the growth of infections in aquaculture.
References
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Ahmad, B., Shah, M., & Choi, S. 2019. Oceans as a source of immunotherapy. Marine Drugs, 17(282), 1-37. https://doi.org/10.3390/md17050282
Ahmed, I., Asgher, M., Sher, F., Hussain, S.M., Nazish, N., Joshi, N., Sharma, A., Parra-Saldívar, R., Bilal, M., & Iqbal, H.M.N. (2022). Exploring marine as a rich source of bioactive peptides: challenges and opportunities from marine pharmacology. Marine Drugs, 20(208). https://doi.org/10.3390/md20030208
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Angelini, P., Abdel-Azeem, A., & Mand-Girometta, C.E. (2022). Editorial: Bioactive compounds with potential medicinal properties derived from fungi: recent and future developments in microbial biotechnology. Frontiers in Microbiology, 13: 1-3. https://doi.org/10.3389/fmicb.2022.837586
Artanti, N., Tachibana, S., Kardono, L.B.S., & Sukiman, H. (2011). Screening of endophytic fungi having ability for antioxidant and α-glukosidase inhibitor activities isolated from Taxus sumatrana. Pakistan Journal of Biological Sciences, 14(22), 1019-1023.
Asaf, R., Athirah, A., & Paena, M. (2021). Optimalisasi pengembangan usaha budi daya rumput laut (Kappaphycus alvarezii) di Perairan Teluk Kulisusu Kabupaten Buton Utara Provinsi Sulawesi Tenggara. Jurnal Sosial Ekonomi Kelautan dan Perikanan, 16(1), 39-50.
Askari, H., Khajepour, A., Khamesee, M. B., Saadatnia, Z., & Wang, Z. L. (2018). Piezoelectric and triboelectric nanogenerators: Trends and impacts. Nano Today, 22, 10-13
Bao, J., Sun, L., Zhang, X., & Liu, H. W. (2020). Terrein: a bioactive metabolite with therapeutic potential. Marine Drugs, 18(11), 543.
Davis & Stout. (1971). Disc Plate Method of Microbiological Antibiotic Essay. Journal Of Microbiology, 22(4), 39.
Ekasari, J., Achmadi, F. F., Isti’anah, I., & Tarman, K. (2023). Growth performance and blood figures of African catfish (Clarias sp.) fed with marine fungal fermented feed. In IOP Conference Series: Earth and Environmental Science (Vol. 1137, No. 1, p. 012041). IOP Publishing.
Guo, L., Zhang, F., Wang, X., Chen, H., Wang, Q., Guo, J., Cao, X., & Wang, L. (2019). Antibacterial activity and action mechanism of questin from marine Aspergillus flavipes HN4-13 against aquatic pathogen Vibrio harveyi. 3 Biotech, 9, 1-7.
Hakim, S. K., Sudirman, S., Janna, M., Nugroho, G. D., & Sari, D. I. (2024). Aktivitas antibakteri Pseudomonas aeruginosa dan Staphylococcus aureus dari ekstrak etanol selada air (Nasturtium officinale). Jurnal Pengolahan Hasil Perikanan Indonesia, 27(4), 319-326. http://dx.doi.org/10.17844/jphpi.v27i4.50254
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Imra, Tarman, K., & Desniar. (2016). Aktivitas antioksidan dan antibakteri nipah (Nypa fruticans) terhadap Vibrio sp. isolat kepiting bakau (Scylla sp.). Jurnal Pengolahan Hasil Perikanan Indonesia, 19(3), 241-250.
Isti’anah, I., Budiman, M. A., Suseno, S. H., Nugraha, R., Effendi, I., & Tarman, K. (2024). Action Mechanism of Marine Endophytic Fungi Aspergillus terreus as Antibacterial Agent Against Vibrio harveyi. In BIO Web of Conferences (Vol. 92, p. 02008). EDP Sciences.
Isti’anah, I., Mubarik, N. R., & Tjahjoleksono, A. (2021). Characterization and application of nitrogen-fixing and indole-3-acetic acid producing bacteria A13 in Oil Palm (Elaeisguineensis Jacq.) seedling. Journal of Microbial Systematics and Biotechnology, 3(1), 32-40.
Jalil, M. T. M., Zakaria, N. A., Suhaimi, N. S. M., & Ibrahim, D. (2022). Crude extracts of an endophytic fungus attenuate the growth of pathogenic bacteria in aquaculture. Iranian Journal of Microbiology, 14(3), 383-394.
Juliana, M., & Yulian, M. (2020). Identifikasi kloramfenikol pada udang vaname (Litopenaeus vannamei) menggunakan high performance Liquid chromatography (hplc). AMINA, 2(1), 13-18.
Julianti, E., Singgih, M., Ikram, M.R., Naufal, A., Putra, M.Y., & Hadi, T.A. (2019, Oktober 24-26). Antimicrobial activity of fungi isolated from the marine sponges collected from Sekotong Beach Lombok, Indonesia. [Conference session] The First Maluku International Conference on Marine Science and Technology 2018, Ambon, Indonesia. IOP Conf. Series: Earth and Environmental Science, 339, doi:10.1088/1755-1315/339/1/012058.
Kaliu, S., & Fitra, R. A. (2019). Komposisi vegetasi mangrove dan identifikasi fauna secara vertikal di Pesisir Pantai Kecamatan Pomala Sulawesi Tenggara. SAINTIFIK: Jurnal Matematika, Sains, dan Pembelajarannya, 5(2), 127-134.
Karthikeyan, A., Joseph, A., & Nair, B.G. (2022). Promising bioactive compounds from the marine environment and their potential effects on various diseases. Journal of Genetic Engineering and Biotechnology, 20(14), 1-38
Laksananny, S. A., Poedjirahajoe, E., Purwanto, R. H., & Hermawan, M. T. T. (2020). Analisis vegetasi hutan mangrove di Kabupaten Buton Utara (Studi kasus di Kecamatan Kulisusu Barat, Kabupaten Buton Utara, Sulawesi Tenggara). Jurnal Ilmu Lingkungan, 18(3), 515-521.
Li, Q., Han, W., Yang, C., Si, Y., Xin, M., Guan, H., & Li, C. (2022). Low molecular-weight polyguluronate phosphate: An immunostimulant by activating splenocyte/macrophage in vitro and improving immune response in vivo. International Journal of Biological Macromolecules, 216, 510-519.
Mahmoud MM, Abdel-Razek AS, Soliman HSM, Ponomareva LV, Thorson JS, Shaaban KA, Shaaban M. (2021). Diverse polyketides from the marine endophytic Alternaria sp. LV52: Structure determination and cytotoxic activities. Biotechnology Reports 33: 1-7. https://doi.org/10.1016/j.btre.2021.e00628
Morya, R., & Tanna, B. (2014). Antibacterial potential of marine derived fungi against Vibrio harveyi: a causative agent of white spot syndrome in shrimp. Aquaculture, 434, 186-190.
Mukherjee, G., Singh, Y., & Sen, S. K. (2021). In vitro cytotoxicity of citrinin metabolites from Aspergillus terreus. Journal of Applied Pharmaceutical Science, 11(02), 098-103.
Mursit, A., Wahyono, A., & Setiawan, Y. (2022). Strategi peningkatan ekspor produk kelautan dan perikanan ke pasar Eropa. Jurnal Manajemen USNI, 6(2), 9-24.
Papuangan, N. (2009). Aktivitas penghambatan senyawa antimikrob Streptomyces spp. terhadap mikrob patogen tular tanah secara in vitro dan in planta [Tesis]. Institut Pertanian Bogor.
Prihanto, A. A. (2012). Perbandingan aktivitas antibakteri Penicillium notatum ATCC 28089 dengan Penicillium sp. R1M yang diisolasi dari mangrove Sonneratia caseolaris. Jurnal Pengolahan Hasil Perikanan Indonesia, 15(1): 66-70. https://doi.org/10.17844/jphpi.v15i1.5336
Rahaweman, A.C, Pamungkas, J., Madduppa, H., Thoms, C., & Tarman, K. (2016, September 12-13). Screening of endophytic fungi from chlorophyta and phaeophyta for antibacterial activity. The 4th International Conference on ICMMBT 2023, Bogor, Indonesia. IOP. Conf. Series: Earth and Environmental Science, 31, 1-7.
Rochvita, A., & Permadi J. (2021). Agribisnis perikanan new normal: kajian strategi recovery usaha perikanan budidaya masa darurat Covid-19 di Kabupaten Magelang. AGRIKAN: Jurnal Agribisnis Perikanan, 14(2), 622-632.
Ruiz, B., Chávez, A., Forero, A., GarcíaHuante, Y., Romero, A., & Sánchez, M. (2010). Production of microbial secondary metabolites: regulation by the carbon source. Critical. Reviews in Microbiology, 36: 146–167.
Sarasan, M., Puthumana, J., Job, N., Han, J., Lee, J., & Philip, R. (2017). Marine algicolous endophytic fungi - a promising drug resource of the era. J. Microbiol. Biotechnology, 27(6), 1039-1052.
Saputri, M. P., Utami, R., Fadhila, J., & Handayani, S. N. (2019). Uji aktivitas antibakteri dan antiinflamasi ekstrak daun Ageratum conyzoides L (Bandotan). Prosiding Program Kreativitas Mahasiswa, 21.
Srinivasan, N., Thangavelu, K., & Uthandi, S. (2022). Lovastatin production by an oleaginous fungus, Aspergillus terreus KPR12 using sago processing wastewater (SWW). Microbial Cell Factories, 21(1), 1-14.
Tarman K. (2011). Biological and chemical investigations of Indonesian marine derived fungi and their secondary metabolites. Cuvillier Verlag.
Tarman, K., Purwaningsih, S., & Negara A. A. P. A. P. (2013). Aktivitas antibakteri ekstrak daun bakau hitam (Rhizophora mucronata) terhadap bakteri penyebab diare. Jurnal Pengolahan Hasil Perikanan Indonesia, 16(3), 249-258. https://doi.org/10.17844/jphpi.v16i3.8063
Torta, L. Burruano, S.; Giambra, S.; Conigliaro, G.; Piazza, G.; Mirabile, G.; Pirrotta, M.; Calvo, R.; Bellissimo, G.; Calvo, S. (2022). Cultivabl fungal endophytes in roots, rhizomes and leaves of Posidonia oceanica (L.) delile along the coast of Sicily, Italy. Plants,11, 1139. https://doi.org/10.3390/plants11091139
Umesha, S., Manukumar, H. M., & Raghava, S. (2016). A rapid method for isolation of genomic DNA from food-borne fungal pathogens. 3 Biotech, 6, 1-9.
Wahjuningrum, D., Efianda, T. R., Tarman, K., Yuhana, M., Effendi, I., & Saputra, F. (2020). Supplementation of Nodulisporium sp. KT29 induced by Vibrio harveyi as an immunostimulant for controlling vibriosis in vannamei white shrimp under marine culture system. Jurnal Akuakultur Indonesia, 19(2), 95-105.
Wahyuni S.H., Hasanuddin, dan Purba E. (2016). Identifikasi dan antagonisme jamur endofit tanaman tebu (Saccharum officinarum L.) dalam menghambat Xanthomonas albilineans L. penyebab penyakit vaskular bakteri. Jurnal Pertanian Tropik, 3(1), 31-42.
Abd El-latif, H. H., Hassan, S. W., & Beltagy, E. A. (2021). Optimization and characterization of antimicrobial agents produced by Marine Aspergillus terreus SHE05 against Aeromonas hydrophila and other Applications. Journal of Pure and Applied Microbiology, 15(4), 2367-2382.
Ahmad, B., Shah, M., & Choi, S. 2019. Oceans as a source of immunotherapy. Marine Drugs, 17(282), 1-37. https://doi.org/10.3390/md17050282
Ahmed, I., Asgher, M., Sher, F., Hussain, S.M., Nazish, N., Joshi, N., Sharma, A., Parra-Saldívar, R., Bilal, M., & Iqbal, H.M.N. (2022). Exploring marine as a rich source of bioactive peptides: challenges and opportunities from marine pharmacology. Marine Drugs, 20(208). https://doi.org/10.3390/md20030208
Alisyukur, L.O., Sunarto, S., & Marfai, M. A. (2020). Skenario pengelolaan kepesisiran berkelanjutan Kabupaten Buton Selatan, Provinsi Sulawesi Tenggara. Ecotrophic, 14(2), 100-110.
Angelini, P., Abdel-Azeem, A., & Mand-Girometta, C.E. (2022). Editorial: Bioactive compounds with potential medicinal properties derived from fungi: recent and future developments in microbial biotechnology. Frontiers in Microbiology, 13: 1-3. https://doi.org/10.3389/fmicb.2022.837586
Artanti, N., Tachibana, S., Kardono, L.B.S., & Sukiman, H. (2011). Screening of endophytic fungi having ability for antioxidant and α-glukosidase inhibitor activities isolated from Taxus sumatrana. Pakistan Journal of Biological Sciences, 14(22), 1019-1023.
Asaf, R., Athirah, A., & Paena, M. (2021). Optimalisasi pengembangan usaha budi daya rumput laut (Kappaphycus alvarezii) di Perairan Teluk Kulisusu Kabupaten Buton Utara Provinsi Sulawesi Tenggara. Jurnal Sosial Ekonomi Kelautan dan Perikanan, 16(1), 39-50.
Askari, H., Khajepour, A., Khamesee, M. B., Saadatnia, Z., & Wang, Z. L. (2018). Piezoelectric and triboelectric nanogenerators: Trends and impacts. Nano Today, 22, 10-13
Bao, J., Sun, L., Zhang, X., & Liu, H. W. (2020). Terrein: a bioactive metabolite with therapeutic potential. Marine Drugs, 18(11), 543.
Davis & Stout. (1971). Disc Plate Method of Microbiological Antibiotic Essay. Journal Of Microbiology, 22(4), 39.
Ekasari, J., Achmadi, F. F., Isti’anah, I., & Tarman, K. (2023). Growth performance and blood figures of African catfish (Clarias sp.) fed with marine fungal fermented feed. In IOP Conference Series: Earth and Environmental Science (Vol. 1137, No. 1, p. 012041). IOP Publishing.
Guo, L., Zhang, F., Wang, X., Chen, H., Wang, Q., Guo, J., Cao, X., & Wang, L. (2019). Antibacterial activity and action mechanism of questin from marine Aspergillus flavipes HN4-13 against aquatic pathogen Vibrio harveyi. 3 Biotech, 9, 1-7.
Hakim, S. K., Sudirman, S., Janna, M., Nugroho, G. D., & Sari, D. I. (2024). Aktivitas antibakteri Pseudomonas aeruginosa dan Staphylococcus aureus dari ekstrak etanol selada air (Nasturtium officinale). Jurnal Pengolahan Hasil Perikanan Indonesia, 27(4), 319-326. http://dx.doi.org/10.17844/jphpi.v27i4.50254
Holo, H., Nilssen, & Ness, I. F. (1991). Lactococcin A, a new bacteriocin from Lactococcus lactis subsp. cremoris: isolation and characterization of the protein and its gene. Journal of Bacteriology, 38 (12):79-87.
Imra, Tarman, K., & Desniar. (2016). Aktivitas antioksidan dan antibakteri nipah (Nypa fruticans) terhadap Vibrio sp. isolat kepiting bakau (Scylla sp.). Jurnal Pengolahan Hasil Perikanan Indonesia, 19(3), 241-250.
Isti’anah, I., Budiman, M. A., Suseno, S. H., Nugraha, R., Effendi, I., & Tarman, K. (2024). Action Mechanism of Marine Endophytic Fungi Aspergillus terreus as Antibacterial Agent Against Vibrio harveyi. In BIO Web of Conferences (Vol. 92, p. 02008). EDP Sciences.
Isti’anah, I., Mubarik, N. R., & Tjahjoleksono, A. (2021). Characterization and application of nitrogen-fixing and indole-3-acetic acid producing bacteria A13 in Oil Palm (Elaeisguineensis Jacq.) seedling. Journal of Microbial Systematics and Biotechnology, 3(1), 32-40.
Jalil, M. T. M., Zakaria, N. A., Suhaimi, N. S. M., & Ibrahim, D. (2022). Crude extracts of an endophytic fungus attenuate the growth of pathogenic bacteria in aquaculture. Iranian Journal of Microbiology, 14(3), 383-394.
Juliana, M., & Yulian, M. (2020). Identifikasi kloramfenikol pada udang vaname (Litopenaeus vannamei) menggunakan high performance Liquid chromatography (hplc). AMINA, 2(1), 13-18.
Julianti, E., Singgih, M., Ikram, M.R., Naufal, A., Putra, M.Y., & Hadi, T.A. (2019, Oktober 24-26). Antimicrobial activity of fungi isolated from the marine sponges collected from Sekotong Beach Lombok, Indonesia. [Conference session] The First Maluku International Conference on Marine Science and Technology 2018, Ambon, Indonesia. IOP Conf. Series: Earth and Environmental Science, 339, doi:10.1088/1755-1315/339/1/012058.
Kaliu, S., & Fitra, R. A. (2019). Komposisi vegetasi mangrove dan identifikasi fauna secara vertikal di Pesisir Pantai Kecamatan Pomala Sulawesi Tenggara. SAINTIFIK: Jurnal Matematika, Sains, dan Pembelajarannya, 5(2), 127-134.
Karthikeyan, A., Joseph, A., & Nair, B.G. (2022). Promising bioactive compounds from the marine environment and their potential effects on various diseases. Journal of Genetic Engineering and Biotechnology, 20(14), 1-38
Laksananny, S. A., Poedjirahajoe, E., Purwanto, R. H., & Hermawan, M. T. T. (2020). Analisis vegetasi hutan mangrove di Kabupaten Buton Utara (Studi kasus di Kecamatan Kulisusu Barat, Kabupaten Buton Utara, Sulawesi Tenggara). Jurnal Ilmu Lingkungan, 18(3), 515-521.
Li, Q., Han, W., Yang, C., Si, Y., Xin, M., Guan, H., & Li, C. (2022). Low molecular-weight polyguluronate phosphate: An immunostimulant by activating splenocyte/macrophage in vitro and improving immune response in vivo. International Journal of Biological Macromolecules, 216, 510-519.
Mahmoud MM, Abdel-Razek AS, Soliman HSM, Ponomareva LV, Thorson JS, Shaaban KA, Shaaban M. (2021). Diverse polyketides from the marine endophytic Alternaria sp. LV52: Structure determination and cytotoxic activities. Biotechnology Reports 33: 1-7. https://doi.org/10.1016/j.btre.2021.e00628
Morya, R., & Tanna, B. (2014). Antibacterial potential of marine derived fungi against Vibrio harveyi: a causative agent of white spot syndrome in shrimp. Aquaculture, 434, 186-190.
Mukherjee, G., Singh, Y., & Sen, S. K. (2021). In vitro cytotoxicity of citrinin metabolites from Aspergillus terreus. Journal of Applied Pharmaceutical Science, 11(02), 098-103.
Mursit, A., Wahyono, A., & Setiawan, Y. (2022). Strategi peningkatan ekspor produk kelautan dan perikanan ke pasar Eropa. Jurnal Manajemen USNI, 6(2), 9-24.
Papuangan, N. (2009). Aktivitas penghambatan senyawa antimikrob Streptomyces spp. terhadap mikrob patogen tular tanah secara in vitro dan in planta [Tesis]. Institut Pertanian Bogor.
Prihanto, A. A. (2012). Perbandingan aktivitas antibakteri Penicillium notatum ATCC 28089 dengan Penicillium sp. R1M yang diisolasi dari mangrove Sonneratia caseolaris. Jurnal Pengolahan Hasil Perikanan Indonesia, 15(1): 66-70. https://doi.org/10.17844/jphpi.v15i1.5336
Rahaweman, A.C, Pamungkas, J., Madduppa, H., Thoms, C., & Tarman, K. (2016, September 12-13). Screening of endophytic fungi from chlorophyta and phaeophyta for antibacterial activity. The 4th International Conference on ICMMBT 2023, Bogor, Indonesia. IOP. Conf. Series: Earth and Environmental Science, 31, 1-7.
Rochvita, A., & Permadi J. (2021). Agribisnis perikanan new normal: kajian strategi recovery usaha perikanan budidaya masa darurat Covid-19 di Kabupaten Magelang. AGRIKAN: Jurnal Agribisnis Perikanan, 14(2), 622-632.
Ruiz, B., Chávez, A., Forero, A., GarcíaHuante, Y., Romero, A., & Sánchez, M. (2010). Production of microbial secondary metabolites: regulation by the carbon source. Critical. Reviews in Microbiology, 36: 146–167.
Sarasan, M., Puthumana, J., Job, N., Han, J., Lee, J., & Philip, R. (2017). Marine algicolous endophytic fungi - a promising drug resource of the era. J. Microbiol. Biotechnology, 27(6), 1039-1052.
Saputri, M. P., Utami, R., Fadhila, J., & Handayani, S. N. (2019). Uji aktivitas antibakteri dan antiinflamasi ekstrak daun Ageratum conyzoides L (Bandotan). Prosiding Program Kreativitas Mahasiswa, 21.
Srinivasan, N., Thangavelu, K., & Uthandi, S. (2022). Lovastatin production by an oleaginous fungus, Aspergillus terreus KPR12 using sago processing wastewater (SWW). Microbial Cell Factories, 21(1), 1-14.
Tarman K. (2011). Biological and chemical investigations of Indonesian marine derived fungi and their secondary metabolites. Cuvillier Verlag.
Tarman, K., Purwaningsih, S., & Negara A. A. P. A. P. (2013). Aktivitas antibakteri ekstrak daun bakau hitam (Rhizophora mucronata) terhadap bakteri penyebab diare. Jurnal Pengolahan Hasil Perikanan Indonesia, 16(3), 249-258. https://doi.org/10.17844/jphpi.v16i3.8063
Torta, L. Burruano, S.; Giambra, S.; Conigliaro, G.; Piazza, G.; Mirabile, G.; Pirrotta, M.; Calvo, R.; Bellissimo, G.; Calvo, S. (2022). Cultivabl fungal endophytes in roots, rhizomes and leaves of Posidonia oceanica (L.) delile along the coast of Sicily, Italy. Plants,11, 1139. https://doi.org/10.3390/plants11091139
Umesha, S., Manukumar, H. M., & Raghava, S. (2016). A rapid method for isolation of genomic DNA from food-borne fungal pathogens. 3 Biotech, 6, 1-9.
Wahjuningrum, D., Efianda, T. R., Tarman, K., Yuhana, M., Effendi, I., & Saputra, F. (2020). Supplementation of Nodulisporium sp. KT29 induced by Vibrio harveyi as an immunostimulant for controlling vibriosis in vannamei white shrimp under marine culture system. Jurnal Akuakultur Indonesia, 19(2), 95-105.
Wahyuni S.H., Hasanuddin, dan Purba E. (2016). Identifikasi dan antagonisme jamur endofit tanaman tebu (Saccharum officinarum L.) dalam menghambat Xanthomonas albilineans L. penyebab penyakit vaskular bakteri. Jurnal Pertanian Tropik, 3(1), 31-42.
Authors
Isti’anah I., Tarman K., Suseno S. H., Nugraha R., & Effendi I. (2024). Penapisan senyawa bioaktif antibakteri fungi laut endofit asal Pulau Buton Sulawesi Tenggara: Screening bioactive compounds of marine endophytic fungi as antibacteria from Buton Island, Southeast Sulawesi . Jurnal Pengolahan Hasil Perikanan Indonesia, 27(7). https://doi.org/10.17844/jphpi.v27i7.50489
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