Research Article
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The Co-Isolation of Lactic Acid Bacteria (LAB) and A Related Pathogenic Strain from Pangasius Nasutus

Year 2023, Volume: 6 Issue: 2, 143 - 154, 15.08.2023
https://doi.org/10.38001/ijlsb.1268388

Abstract

Catfish species Pangasius nasutus, or locally known as Patin Buah is one of a popular aquaculture product in Malaysia. Probiotic strain is an attractive alternative to conventional antibiotics in managing aquaculture diseases. Pangasius can be a source of bacterial strain in developing suitable probiotic useful in Pangasius rearing. This study highlights on the isolation of LAB strains from the selected tissues from P. nasutus which are the heart, stomach, and intestines. These strains were subjected to morphological, biochemical, and genotypic characterisations. Three different strains were isolated from fish tissues, H-Hn from heart; S-Hn from stomach; and I-Sk from intestine. Biochemical characterisation were consistent with ribosomal rRNA sequencing, in which strain H-Hn (acc. No. MW504962) is highly similar (100%) to Lactococcus lactis, and both I-Sk (MW504964) and S-Hn (MW504963) are similar (99%) to Lactococcus garvieae. All strains appeared to be non-spore forming, non-motile, Gram-positive coccus, catalase and citrate negative, and fermenting sugar. Unlike H-Hn, both H-Sk and I-Sk strains were able to grow at 6.5% NaCl and at higher temperature (45°C). The antimicrobial properties were assessed using agar disk diffusion assay against several indicator organisms (Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus and Vibrio parahaemolyticus). These strains exhibited varying degree of inhibitory properties against the indicator organisms. Strain H-Hn inhibited E. coli and V. parahaemolyticus. Meanwhile, strains I-Sk and S-Hn showed a rather broad-spectrum inhibition. From P. nasutus, other L. lactis strain, a pathogenic species from L. garvieae could also be found especially in their gut tissues. The presence of L. garvieae in commercial P. nasutus should raise some concern to those who like to consume this fish. Meanwhile, the LAB strains isolated has probiotics potential that can be commercially used in the managing fish diseases in aquaculture.

Supporting Institution

International Islamic University Malaysia

Project Number

SDG 2020-2021 -Patin Buah as a sustainable food source flagship project (IIUM)

Thanks

Thank to international Islamic University Malaysia for partially funding this project

References

  • 1. Chiew, M.K.I., A.M. Salter, and Y.Z. Yin Sze Lim, The significance of major viral and bacterial diseases in Malaysian aquaculture industry. Pertanika Journal of Tropical Agricultural Science, 2019. 42(3): p. 1023-1047.
  • 2. Alagesh, T., 'Ikan patin' at risk of becoming endangered. , in New Straits Time. 2017, NST Press (M) Sdn, Bhd.
  • 3. Preena, P.G., et al., Antimicrobial resistance in aquaculture: a crisis for concern. Biologia, 2020. 75(9): p. 1497-1517.
  • 4. Ee Lean, T., et al., Antibiotic residues from aquaculture farms and their ecological risks in Southeast Asia: a case study from Malaysia. Ecosystem Health and Sustainability, 2021. 7(1): p. 1926337.
  • 5. Wang, C., et al., Beneficial bacteria for aquaculture: nutrition, bacteriostasis and immunoregulation. J Appl Microbiol, 2020. 128(1): p. 28-40.
  • 6. Anjana and S. Tiwari, Bacteriocin-Producing Probiotic Lactic Acid Bacteria in Controlling Dysbiosis of the Gut Microbiota. Frontiers in Cellular and Infection Microbiology, 2022. 12.
  • 7. Hien Van, D., et al., Host-Associated Probiotics: A Key Factor in Sustainable Aquaculture. Reviews in Fisheries Science & Aquaculture, 2019. 28(1): p. 16-42.
  • 8. Pereira, W.A., et al., Bacteriocinogenic probiotic bacteria isolated from an aquatic environment inhibit the growth of food and fish pathogens. Scientific Reports, 2022. 12(1): p. 5530.
  • 9. Goa, T., et al., Isolation and Characterization of Lactic Acid Bacteria from Fermented Milk Produced in Jimma Town, Southwest Ethiopia, and Evaluation of their Antimicrobial Activity against Selected Pathogenic Bacteria. Int J Food Sci, 2022. 2022: p. 2076021.
  • 10. Wambui, J., P. Lamuka, and P. Njage, Lactic acid bacteria isolates from fermented camel milk (suusac) are potential protective cultures of raw camel meat. International Journal of Agriculture and Environmental Research, 2017. 3: p. 2960-2975.
  • 11. Sandra, T.-S., et al., In vitro evaluation of the antimicrobial activity of a range of probiotics against pathogens: Evidence for the effects of organic acids. Anaerobe, 2012. 18(5): p. 530-538.
  • 12. Diop, M.B., et al., Bacteriocin producers from traditional food products. Biotechnologie, Agronomie, Société et Environnement, 2007. 11(4): p. 275.
  • 13. Hamid, T.T.A.H. and N.F.A.M. Fuzi, Lactic acid bacterium with antimicrobial properties from selected malay traditional fermented foods. International Journal of Life Sciences and Biotechnology, 2021. 4(1-2): p. 11-34.
  • 14. Lemoine, F., Correia, D., Lefort, V., Doppelt-Azeroual, O., Mareuil, F., Cohen-Boulakia, S. , & Gascuel, O., NGPhylogeny.fr: new generation phylogenetic services for non-specialists. Nucleic Acids Research, 47(W1), W260-W265. Nucleic Acids Research, , 2019. 47(W1): p. W260-W265.
  • 15. Pulipati, S., et al., Fermented veggies: antibacterial efficacy of isolated lactic acid bacteria. World Journal of Pharmacy and Pharmaceutical Sciences, 2019. 8(9): p. 270-283.
  • 16. Haro, G., I. Iksen, and N. Nasri, Identification, characterization and antibacterial potential of probiotic lactic acid bacteria isolated from Naniura (a traditional batak fermented food from Carp) against Salmonella typhi. Rasayan Journal of Chemistry, 2020. 13: p. 464-468.
  • 17. Suzuki, A. and M. Suzuki, Antimicrobial Activity of Lactococcus lactis subsp. lactis Isolated from a Stranded Cuvier’s Beaked Whale (Ziphius cavirostris) against Gram-Positive and -Negative Bacteria. . Microorganisms, 2021. 9(2): p. 243.
  • 18. Itoi, S., et al., Isolation of halotolerant Lactococcus lactis subsp. lactis from intestinal tract of coastal fish. Int J Food Microbiol, 2008. 121(1): p. 116-21.
  • 19. Itoi, S., et al., The clam Meretrix lamarckii (Bivalvia: Veneridae) is a rich repository of marine lactic acid bacterial strains. Annals of Microbiology, 2014. 64(3): p. 1267-1274.
  • 20. Cadirci, B. and S. Citak, Antagonistic Effects of Some Lactobacilli On Some Gram-Negative Bacteria. Gazi Univ J Sci, 2010. 23.
  • 21. Noraphat, H., et al., Inhibition of food-spoilage and foodborne pathogenic bacteria by a nisin Z-producing Lactococcus lactis subsp. lactis KT2W2L. LWT - Food Science and Technology, 2017. 82: p. 170-175.
  • 22. Hamid, T.A.T.H., I. Khairil, and A. Mohammed, Lactococcus Lactis Strains from Intestinal Organ of Black Tips Shark Carcharhinus Limbatus Producing Nisin-Like Bacteriocin Active against Shrimp and Fish Pathogens (Vibrio Parahaemolyticus and Vibrio Alginolyticus). Journal of Microbiology, Biotechnology & Food Science, 2020. 10(3).
  • 23. Van Doan, H., et al., Host-associated probiotics boosted mucosal and serum immunity, disease resistance and growth performance of Nile tilapia (Oreochromis niloticus). Aquaculture, 2018. 491: p. 94-100.
  • 24. Meyburgh, C.M., R.R. Bragg, and C.E. Boucher, Lactococcus garvieae: an emerging bacterial pathogen of fish. Dis Aquat Organ, 2017. 123(1): p. 67-79.
  • 25. Malek Alexandre, et al., Lactococcus garvieae, an unusual pathogen in infective endocarditis: case report and review of the literature. BMC Infectious Diseases, 2019. 19(1): p. 301.
  • 26. Varsha, K.K. and K.M. Nampoothiri, Lactococcus garvieae subsp. bovis subsp. nov., lactic acid bacteria isolated from wild gaur (Bos gaurus) dung, and description of Lactococcus garvieae subsp. garvieae subsp. nov. Int J Syst Evol Microbiol, 2016. 66(10): p. 3805-3809.
  • 27. Vendrell, D., Balcázar, J. L., Ruiz-Zarzuela, I., de Blas, I., Gironés, O., & Múzquiz, J. L., Lactococcus garvieae in fish: a review. Comparative immunology, microbiology and infectious diseases,. 2006. 29(4): p. 177–198.
  • 28. Soltani, M., et al., Lactococcosis a re-Emerging disease in aquaculture: disease significant and phytotherapy. Vet Sci, 2021. 8(9).
  • 29. Ismail, S., M.S. Kamarudin, and E. Ramezani-Fard, Performance of Commercial Poultry Offal Meal as Fishmeal Replacement in the Diet of Juvenile Malaysian Mahseer, Tor tambroides. Asian Journal of Animal and Veterinary Advances, 2013. 8: p. 284-292.

Laktik Asit Bakterilerinin (LAB) ve Pangasius Nasutus'tan İlgili Bir Patojenik Suşun Birlikte İzolasyonu

Year 2023, Volume: 6 Issue: 2, 143 - 154, 15.08.2023
https://doi.org/10.38001/ijlsb.1268388

Abstract

Catfish species Pangasius nasutus, or locally known as Patin Buah is one of a popular aquaculture product in Malaysia. Probiotic strain is an attractive alternative to conventional antibiotics in managing aquaculture diseases. Pangasius can be a source of bacterial strain in developing suitable probiotic useful in Pangasius rearing. This study highlights on the isolation of LAB strains from the selected tissues from P. nasutus which are the heart, stomach, and intestines. These strains were subjected to morphological, biochemical, and genotypic characterisations. Three different strains were isolated from fish tissues, H-Hn from heart; S-Hn from stomach; and I-Sk from intestine. Biochemical characterisation were consistent with ribosomal rRNA sequencing, in which strain H-Hn (acc. No. MW504962) is highly similar (100%) to Lactococcus lactis, and both I-Sk (MW504964) and S-Hn (MW504963) are similar (99%) to Lactococcus garvieae. All strains appeared to be non-spore forming, non-motile, Gram-positive coccus, catalase and citrate negative, and fermenting sugar. Unlike H-Hn, both H-Sk and I-Sk strains were able to grow at 6.5% NaCl and at higher temperature (45°C). The antimicrobial properties were assessed using agar disk diffusion assay against several indicator organisms (Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus and Vibrio parahaemolyticus). These strains exhibited varying degree of inhibitory properties against the indicator organisms. Strain H-Hn inhibited E. coli and V. parahaemolyticus. Meanwhile, strains I-Sk and S-Hn showed a rather broad-spectrum inhibition. From P. nasutus, other L. lactis strain, a pathogenic species from L garvieae could also be found especially in their gut tissues. The presence of L. garvieae in commercial P. nasutus should raise some concern to those who like to consume this fish. Meanwhile, the LAB strains isolated has probiotics potential that can be commercially used in the managing fish diseases in aquaculture.

Project Number

SDG 2020-2021 -Patin Buah as a sustainable food source flagship project (IIUM)

References

  • 1. Chiew, M.K.I., A.M. Salter, and Y.Z. Yin Sze Lim, The significance of major viral and bacterial diseases in Malaysian aquaculture industry. Pertanika Journal of Tropical Agricultural Science, 2019. 42(3): p. 1023-1047.
  • 2. Alagesh, T., 'Ikan patin' at risk of becoming endangered. , in New Straits Time. 2017, NST Press (M) Sdn, Bhd.
  • 3. Preena, P.G., et al., Antimicrobial resistance in aquaculture: a crisis for concern. Biologia, 2020. 75(9): p. 1497-1517.
  • 4. Ee Lean, T., et al., Antibiotic residues from aquaculture farms and their ecological risks in Southeast Asia: a case study from Malaysia. Ecosystem Health and Sustainability, 2021. 7(1): p. 1926337.
  • 5. Wang, C., et al., Beneficial bacteria for aquaculture: nutrition, bacteriostasis and immunoregulation. J Appl Microbiol, 2020. 128(1): p. 28-40.
  • 6. Anjana and S. Tiwari, Bacteriocin-Producing Probiotic Lactic Acid Bacteria in Controlling Dysbiosis of the Gut Microbiota. Frontiers in Cellular and Infection Microbiology, 2022. 12.
  • 7. Hien Van, D., et al., Host-Associated Probiotics: A Key Factor in Sustainable Aquaculture. Reviews in Fisheries Science & Aquaculture, 2019. 28(1): p. 16-42.
  • 8. Pereira, W.A., et al., Bacteriocinogenic probiotic bacteria isolated from an aquatic environment inhibit the growth of food and fish pathogens. Scientific Reports, 2022. 12(1): p. 5530.
  • 9. Goa, T., et al., Isolation and Characterization of Lactic Acid Bacteria from Fermented Milk Produced in Jimma Town, Southwest Ethiopia, and Evaluation of their Antimicrobial Activity against Selected Pathogenic Bacteria. Int J Food Sci, 2022. 2022: p. 2076021.
  • 10. Wambui, J., P. Lamuka, and P. Njage, Lactic acid bacteria isolates from fermented camel milk (suusac) are potential protective cultures of raw camel meat. International Journal of Agriculture and Environmental Research, 2017. 3: p. 2960-2975.
  • 11. Sandra, T.-S., et al., In vitro evaluation of the antimicrobial activity of a range of probiotics against pathogens: Evidence for the effects of organic acids. Anaerobe, 2012. 18(5): p. 530-538.
  • 12. Diop, M.B., et al., Bacteriocin producers from traditional food products. Biotechnologie, Agronomie, Société et Environnement, 2007. 11(4): p. 275.
  • 13. Hamid, T.T.A.H. and N.F.A.M. Fuzi, Lactic acid bacterium with antimicrobial properties from selected malay traditional fermented foods. International Journal of Life Sciences and Biotechnology, 2021. 4(1-2): p. 11-34.
  • 14. Lemoine, F., Correia, D., Lefort, V., Doppelt-Azeroual, O., Mareuil, F., Cohen-Boulakia, S. , & Gascuel, O., NGPhylogeny.fr: new generation phylogenetic services for non-specialists. Nucleic Acids Research, 47(W1), W260-W265. Nucleic Acids Research, , 2019. 47(W1): p. W260-W265.
  • 15. Pulipati, S., et al., Fermented veggies: antibacterial efficacy of isolated lactic acid bacteria. World Journal of Pharmacy and Pharmaceutical Sciences, 2019. 8(9): p. 270-283.
  • 16. Haro, G., I. Iksen, and N. Nasri, Identification, characterization and antibacterial potential of probiotic lactic acid bacteria isolated from Naniura (a traditional batak fermented food from Carp) against Salmonella typhi. Rasayan Journal of Chemistry, 2020. 13: p. 464-468.
  • 17. Suzuki, A. and M. Suzuki, Antimicrobial Activity of Lactococcus lactis subsp. lactis Isolated from a Stranded Cuvier’s Beaked Whale (Ziphius cavirostris) against Gram-Positive and -Negative Bacteria. . Microorganisms, 2021. 9(2): p. 243.
  • 18. Itoi, S., et al., Isolation of halotolerant Lactococcus lactis subsp. lactis from intestinal tract of coastal fish. Int J Food Microbiol, 2008. 121(1): p. 116-21.
  • 19. Itoi, S., et al., The clam Meretrix lamarckii (Bivalvia: Veneridae) is a rich repository of marine lactic acid bacterial strains. Annals of Microbiology, 2014. 64(3): p. 1267-1274.
  • 20. Cadirci, B. and S. Citak, Antagonistic Effects of Some Lactobacilli On Some Gram-Negative Bacteria. Gazi Univ J Sci, 2010. 23.
  • 21. Noraphat, H., et al., Inhibition of food-spoilage and foodborne pathogenic bacteria by a nisin Z-producing Lactococcus lactis subsp. lactis KT2W2L. LWT - Food Science and Technology, 2017. 82: p. 170-175.
  • 22. Hamid, T.A.T.H., I. Khairil, and A. Mohammed, Lactococcus Lactis Strains from Intestinal Organ of Black Tips Shark Carcharhinus Limbatus Producing Nisin-Like Bacteriocin Active against Shrimp and Fish Pathogens (Vibrio Parahaemolyticus and Vibrio Alginolyticus). Journal of Microbiology, Biotechnology & Food Science, 2020. 10(3).
  • 23. Van Doan, H., et al., Host-associated probiotics boosted mucosal and serum immunity, disease resistance and growth performance of Nile tilapia (Oreochromis niloticus). Aquaculture, 2018. 491: p. 94-100.
  • 24. Meyburgh, C.M., R.R. Bragg, and C.E. Boucher, Lactococcus garvieae: an emerging bacterial pathogen of fish. Dis Aquat Organ, 2017. 123(1): p. 67-79.
  • 25. Malek Alexandre, et al., Lactococcus garvieae, an unusual pathogen in infective endocarditis: case report and review of the literature. BMC Infectious Diseases, 2019. 19(1): p. 301.
  • 26. Varsha, K.K. and K.M. Nampoothiri, Lactococcus garvieae subsp. bovis subsp. nov., lactic acid bacteria isolated from wild gaur (Bos gaurus) dung, and description of Lactococcus garvieae subsp. garvieae subsp. nov. Int J Syst Evol Microbiol, 2016. 66(10): p. 3805-3809.
  • 27. Vendrell, D., Balcázar, J. L., Ruiz-Zarzuela, I., de Blas, I., Gironés, O., & Múzquiz, J. L., Lactococcus garvieae in fish: a review. Comparative immunology, microbiology and infectious diseases,. 2006. 29(4): p. 177–198.
  • 28. Soltani, M., et al., Lactococcosis a re-Emerging disease in aquaculture: disease significant and phytotherapy. Vet Sci, 2021. 8(9).
  • 29. Ismail, S., M.S. Kamarudin, and E. Ramezani-Fard, Performance of Commercial Poultry Offal Meal as Fishmeal Replacement in the Diet of Juvenile Malaysian Mahseer, Tor tambroides. Asian Journal of Animal and Veterinary Advances, 2013. 8: p. 284-292.
There are 29 citations in total.

Details

Primary Language English
Subjects Structural Biology
Journal Section Research Articles
Authors

Nur Hannah Rashıdah Haja Mohıdeen 0009-0001-1286-3394

Nurul Sakinah Alias 0009-0007-7155-128X

Nur Nazifah Mansur 0000-0002-3615-5851

Tengku Haziyamin Tengku Abdul Hamid 0000-0001-8223-481X

Project Number SDG 2020-2021 -Patin Buah as a sustainable food source flagship project (IIUM)
Early Pub Date July 30, 2023
Publication Date August 15, 2023
Published in Issue Year 2023 Volume: 6 Issue: 2

Cite

EndNote Haja Mohıdeen NHR, Alias NS, Mansur NN, Tengku Abdul Hamid TH (August 1, 2023) The Co-Isolation of Lactic Acid Bacteria (LAB) and A Related Pathogenic Strain from Pangasius Nasutus. International Journal of Life Sciences and Biotechnology 6 2 143–154.

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