Crude extracts of an endophytic fungus attenuate the growth of pathogenic bacteria in aquaculture
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Abstract
Background and Objectives: Endophytic fungi are believed to possess compounds as antibacterial agents. This study was designed to determine in vivo antibacterial activity of the crude extracts from Lasiodiplodia pseudotheobromae IBRL OS-64 against pathogenic bacteria.
Materials and Methods: The qualitative and quantitative screenings were performed using agar plug and disk diffusion antimicrobial tests, respectively. Besides that, the MIC and MBC value of the extracts were determined using broth microdilution assay and morphological changes of the bacterial cells exposed to the extract were observed under Scanning Electron Microscope (SEM).
Results: Agar plug diffusion assay revealed that V. parahaemolyticus ATCC 17802 and Exiguobacterium profundum IBRL MA6 were the most sensitive to the extract with the size of inhibition zones of 11 to ≤ 20 mm. The MIC and MBC values of the extract varied depending on the test bacteria. Observation through SEM revealed that the bacterial cells exposed to the extract experienced severe damage such as irregular shape with crumpled and shrunken cells which led to cell death.
Conclusion: The data suggest that the crude extracts of L. pseudotheobromae IBRL OS-64 exert antibacterial activity against test bacteria and principally affect the cell wall in growing pathogenic bacterial cells.
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27. Balouiri M, Sadiki M, Ibnsouda SK. Methods for in vitro evaluating antimicrobial activity: A review. J Pharm Anal 2016; 6: 71-79.
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30. Lewis K. Persister cells, dormancy and infectious disease. Nat Rev Microbiol 2007; 5: 48-56.31. Pankey GA, Sabath
LD. Clinical relevance of bacteriostatic versus bactericidal mechanisms of action in the treatment of Gram-positive bacterial infections. Clin Infect Dis 2004; 38: 864-870.
32. Parasuraman S. Toxicological screening. J Pharmacol Pharmacother 2011; 2: 74-79.
33. Strobel G, Daisy B. Bioprospecting for microbial endophytes and their natural products. Microbiol Mol Biol Rev 2003; 67: 491-502.
34. Fesel PH, Zuccaro A. Dissecting endophytic lifestyle along the parasitism/mutualism continuum in Arabidopsis. Curr Opin Microbiol 2016; 32: 103-112.
35. Owen NL, Hundley N. Endophytes: the chemical synthesizers inside plants. Sci Prog 2004; 87: 79-99.
36. Pelka M, Danzl C, Distler W, Petschelt A. A new screening test for toxicity testing of dental materials. J Dent 2000; 28: 341-345.
37. Hartmann M, Berditsch M, Hawecker J, Ardakani MF, Gerthsen D, Ulrich AS. Damage of the bacterial cell envelope by antimicrobial peptides Gramicidin S and PGLa as revealed by transmission and scanning electron microscopy. Antimicrob Agents Chemother 2010; 54: 3132-3142.
38. Jeyanthi V, Velusamy P. Anti-methicillin resistant Staphylococcus aureus compound isolation from Halophilic Bacillus amyloliquefaciens MHB1 and determination of its mode of action using electron microscope and flow cytometry analysis. Indian J Microbiol 2016; 56: 148-157.
2. Marudhupandi T, Kumar TTA, Prakash S, Balamurugan J, Dhayanithi NB. Vibrio parahaemolyticus a causative bacterium for tail rot disease in ornamental fish, Amphiprion sebae. Aquac Rep 2017; 8: 39-44.
3. Plant KP, Lapatra SE. Advances in fish vaccine delivery. Dev Comp Immunol 2011; 35: 1256-1262.
4. Soto-Rodriguez SA, Gomez-Gil B, Lozano-Olvera R, Betancourt-Lozano M, Morales-Covarrubias MS. Field and experimental evidence of Vibrio parahaemolyticus as the causative agent of acute hepatopancreatic necrosis disease of cultured shrimp (Litopenaeus vannamei) in Northwestern Mexico. Appl Environ Microbiol 2015; 81: 1689-1699.
5. Goulden EF, Hall MR, Bourne DG, Pereg LL, Hoj L. Pathogenicity and infection cycle of Vibrio owensii in larviculture of the ornate spiny lobster (Panulirus ornatus). Appl Environ Microbiol 2012; 78: 2841-2849.
6. Scharer K, Savioz S, Cernela N, Saegesser G, Stephan R. Occurrence of Vibrio spp. in fish and shellfish Collected from the Swiss market. J Food Prot 2011; 74: 1345-1347.
7. Adam D, Boopathy R. Use of formic acid to control vibriosis in shrimp aquaculture. Biologia 2013; 68: 1017-1021.
8. Jones MK, Oliver JD. Vibrio vulnificus: disease and pathogenesis. Infect Immun 2009; 77: 1723-1733.
9. Chatterjee S, Ghosh R, Mandal NC. Production of bioactive compounds with bactericidal and antioxidant potential by endophytic fungus Alternaria alternata AE1 isolated from Azadirachta indica A. Juss. PLoS One 2019; 14(4):e0214744.
10. Kwon J, Choi O, Kang B, Lee Y, Park J, Kang D, et al. Identification of Lasiodiplodia pseudotheobromae causing mango dieback in Korea. Can J Plant Pathol 2017; 39: 241-245.
11. Rosado AWC, Machado AR, Freire FCO, Pereira OL. Phylogeny, identification and pathogenicity of Lasiodiplodia associated with postharvest stem-end rot of coconut in Brazil. Plant Dis 2016; 100: 561-568.
12. Wei W, Jiang N, Mei YN, Chu YL, Ge HM, Song YC, et al. An antibacterial metabolite from Lasiodiplodia pseudotheobromae F2. Phytochemistry 2014; 100: 103-109.
13. Taufiq MMJ, Darah I. Fungal endophytes isolated from the leaves of a medicinal plant, Ocimum sanctum Linn and evaluation of their antimicrobial activities. Afr J Microbiol Res 2018; 12: 616-622.
14. Ogbole OO, Segun PA, Adeniji AJ. In vitro cytotoxic activity of medicinal plants from Nigeria ethnomedicine on Rhabdomyosarcoma cancer cell line and HPLC analysis of active extracts. BMC Complement Altern Med 2017; 17: 494.
15. Mares D. Electron microscopy of Microsporum cookie after in vitro treatment with protoanemonin: A combined SEM and TEM study. Mycopathologia 1989; 108: 37-46.
16. Dhayanithy G, Subban K, Chelliah J. Diversity and biological activities of endophytic fungi associated with Catharanthus roseus. BMC Microbiol 2019; 19: 22.
17. Hamzah TNT, Lee SY, Hidayat A, Terhem R, Faridah-Hanum I, Mohamed R. Diversity and characterization of endophytic fungi isolated from the tropical mangrove species, Rhizophora mucronata, and identification of potential antagonists against the soil-borne fungus, Fusarium Solani. Front Microbiol 2018; 9: 1707.
18. Jia M, Chen L, Xin HL, Zheng CJ, Rahman K, Han T, et al. A Friendly relationship between endophytic fungi and medicinal plants: a systematic review. Front Microbiol 2016; 7: 906.
19. Beveridge TJ. Structures of Gram-negative cell wall and their derived membrane vesicles. J Bacteriol 1999; 181: 4725-4733.
20. Miller SI, Salama NR. The gram-negative bacterial periplasm: Size matters. PLoS Biol 2018; 16(1):e2004935.
21. Nikaido H. Molecular basis of bacterial outer membrane permeability revisited. Microbiol Mol Biol Rev 2003; 67: 593-656.
22. Poole CF. Thin-layer chromatography: challenges and opportunities. J Chromatogr A 2003; 1000: 963-984.
23. Bag A, Chattopadhyay RR. Evaluation of synergistic antibacterial and antioxidant efficacy of essential oils of spices and herbs in combination. PLoS One 2015; 10(7):e0131321.
24. Marasini BP, Baral P, Aryal P, Ghimire KR, Neupane S, Dahal N, et al. Evaluation of antibacterial activity of some traditionally used medicinal plants against human pathogenic bacteria. Biomed Res Int 2015; 2015: 265425.
25. Wright GD. Bacterial resistance to antibiotics: enzymatic degradation and modification. Adv Drug Deliv Rev 2005; 57: 1451-1470.
26. Olajuyigbe OO, Afolayan AJ. In vitro antibacterial and time-kill evaluation of the Erythrina caffra Thunb. extract against bacteria associated with diarrhoea. ScientificWorldJournal 2012; 2012:738314.
27. Balouiri M, Sadiki M, Ibnsouda SK. Methods for in vitro evaluating antimicrobial activity: A review. J Pharm Anal 2016; 6: 71-79.
28. Wistrand-Yuen E, Knopp M, Hjort K, Koskiniemi S, Berg OG, Andersson DI. Evolution of high-level resistance during low-level antibiotic exposure. Nat Commun 2018; 9: 1599.
29. Nielsen EI, Viberg A, Lowdin E, Cars O, Karisson MO, Sandstrom M. Semimechanistic pharmacokinetic/pharmacodynamic model for assessment of activity of antimicrobial agents for time-kill curve experiments. Antimicrob Agents Chemother 2007; 51: 128-136.
30. Lewis K. Persister cells, dormancy and infectious disease. Nat Rev Microbiol 2007; 5: 48-56.31. Pankey GA, Sabath
LD. Clinical relevance of bacteriostatic versus bactericidal mechanisms of action in the treatment of Gram-positive bacterial infections. Clin Infect Dis 2004; 38: 864-870.
32. Parasuraman S. Toxicological screening. J Pharmacol Pharmacother 2011; 2: 74-79.
33. Strobel G, Daisy B. Bioprospecting for microbial endophytes and their natural products. Microbiol Mol Biol Rev 2003; 67: 491-502.
34. Fesel PH, Zuccaro A. Dissecting endophytic lifestyle along the parasitism/mutualism continuum in Arabidopsis. Curr Opin Microbiol 2016; 32: 103-112.
35. Owen NL, Hundley N. Endophytes: the chemical synthesizers inside plants. Sci Prog 2004; 87: 79-99.
36. Pelka M, Danzl C, Distler W, Petschelt A. A new screening test for toxicity testing of dental materials. J Dent 2000; 28: 341-345.
37. Hartmann M, Berditsch M, Hawecker J, Ardakani MF, Gerthsen D, Ulrich AS. Damage of the bacterial cell envelope by antimicrobial peptides Gramicidin S and PGLa as revealed by transmission and scanning electron microscopy. Antimicrob Agents Chemother 2010; 54: 3132-3142.
38. Jeyanthi V, Velusamy P. Anti-methicillin resistant Staphylococcus aureus compound isolation from Halophilic Bacillus amyloliquefaciens MHB1 and determination of its mode of action using electron microscope and flow cytometry analysis. Indian J Microbiol 2016; 56: 148-157.
| Files | ||
| Issue | Vol 14 No 3 (2022) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v14i3.9780 | |
| Keywords | ||
| Antibacterial agents; Disk diffusion antimicrobial tests; Crude extracts | ||
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How to Cite
1.
Jalil M, Zakaria N, Suhaimi N, Ibrahim D. Crude extracts of an endophytic fungus attenuate the growth of pathogenic bacteria in aquaculture. Iran J Microbiol. 2022;14(3):383-394.
