Isolation of bacteriophage from shrimp ponds in Soc Trang Province for inhibiting biofilm-forming <i>Micrococcus luteus</i>

Tran Huu Hau; Cao Nguyen Thuy Quynh; Thach Thi Hoang Oanh; Nguyen Pham Anh Thi; Truong Thi Bich Van

doi:10.15625/2525-2518/20905

Author affiliations

Authors

Tran Huu Hau \(^1\) Department of Microbial Biotechnology, Institute of Food and Biotechnology, Can Tho University, Campus II, 3/2 Street, Ninh Kieu Ward, Can Tho City, Viet Nam
\(^2\) Department of Agricultural Science and Technology, Faculty of Innovative Agriculture, Fisheries and Food, Prince of Songkla University, Surat Thani Campus, Surat Thani 84000, Thailand https://orcid.org/0009-0001-9234-2694
Cao Nguyen Thuy Quynh \(^1\) Department of Microbial Biotechnology, Institute of Food and Biotechnology, Can Tho University, Campus II, 3/2 Street, Ninh Kieu Ward, Can Tho City, Viet Nam
Thach Thi Hoang Oanh \(^1\) Department of Microbial Biotechnology, Institute of Food and Biotechnology, Can Tho University, Campus II, 3/2 Street, Ninh Kieu Ward, Can Tho City, Viet Nam
Nguyen Pham Anh Thi \(^1\) Department of Microbial Biotechnology, Institute of Food and Biotechnology, Can Tho University, Campus II, 3/2 Street, Ninh Kieu Ward, Can Tho City, Viet Nam
Truong Thi Bich Van \(^1\) Department of Microbial Biotechnology, Institute of Food and Biotechnology, Can Tho University, Campus II, 3/2 Street, Ninh Kieu Ward, Can Tho City, Viet Nam

DOI:

https://doi.org/10.15625/2525-2518/20905

Keywords:

Bacteriophage, biofilm, Micrococcus luteus, Podoviridae, shrimp pond

Abstract

The bacterial biofilm can support several pathogens to be resistant to antibiotics, fungicides, and parasiticides. Slime from shrimp ponds is the common habitat of some biofilm-forming bacteria and their natural enemies, especially bacteriophage. Therefore, this study aimed to isolate these biofilm-forming bacteria from the bottom slime of shrimp ponds and subsequently isolate several bacteriophage strains that were capable of infecting these bacteria. For the methods, the disadvantageous bacteria were collected from the samples and isolated through morphology, biochemical reactions, and molecular biological identification. The bacteriophage was simultaneously collected from the same slime samples and isolated through spot test and double agar overlay assay. Moreover, the bacterial biofilm detachment ability of the bacteriophage was evaluated through the optical density measurement of the bacterial biofilm with and without bacteriophage. As a result, the isolated biofilm-forming bacteria STM1 was both diplococci and tetrad, Gram-positive, non-motile, and yellow colony on the TSA medium. The catalase and oxidase activities of the STM1 were positive, and this strain did not ferment glucose and mannitol. Through morphological observations, biochemical reactions, and 16S rRNA gene analysis, STM1 was similar to Micrococcus luteus (PP495084.1) at 99.5 %. For the bacteriophage selection, four strains detaching bacterial biofilm were isolated at salinities between 5 and 20 ppt, wherein the phage pST9Δ2 strongly detached the STM1 biofilm (peaking at 82 % at 15ppt salinity).

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References

1. Akayli T., Remziye E. Y., Çigdem U. – Diagnosis of Micrococcus luteus infection in cultured sharpsnout sea bream (Diplodus puntazzo Cetti, 1777). J. Int. Sci. Technol., 36 (2020) 214–220.

2. Saleh O., Mohamed M., Abd El-Galil M., El-Kamed A., Sayed H. – Isolation and characterization of Micrococcus luteus from Oreochromis niloticus in Egypt. J. Curr. Vet. Res., 3 (2021) 16–23. https://doi.org/10.21608/jcvr.2021.199412.

3. Pękala A., Paździor E., Antychowicz J., Bernad A., Głowacka H., Więcek B., Niemczuk W. – Kocuria rhizophila and Micrococcus luteus as emerging opportunist pathogens in brown trout (Salmo trutta Linnaeus, 1758) and rainbow trout (Oncorhynchus mykiss Walbaum, 1792). Aquaculture, 486 (2018) 285–289. https://doi.org/10.1016/j.aquaculture.2017.12.028.

4. Geraldine A. R., Rosidah, Herawati H., Bioshina I. B. – Isolation and identification of potential pathogenic bacteria in living carp (Cyprinus carpio Linnaeus, 1758) sold in supermarkets in Cimahi City, Java. World News Nat. Sci., 32 (2020) 21–35.

5. Hillis A. M., Hulse R., Peter P. S. – Methods to isolate possible bacteriophage for Micrococcus luteus and Acinetobacter baumannii. J. Adv. Lab. Res. Biol., 9 (2018) 86–94.

6. Blakeman J. T., Morales-García A. L., Mukherjee J., Gori K., Hayward A. S., Lant N. J., Geoghegan M. – Extracellular DNA provides structural integrity to a Micrococcus luteus biofilm. Langmuir, 35 (2019) 6468–6475. https://doi.org/10.1021/acs.langmuir.9b00297.

7. Gannesen A. V., Ziganshin R. H., Zdorovenko E. L., Klimko A. I., Ianutsevich E. A., Danilova O. A., Tereshina V. M., Gorbachevskii M. V., Ovcharova M. A., Nevolina E. D., et al. – Epinephrine extensively changes the biofilm matrix composition in Micrococcus luteus C01 isolated from human skin. Front. Microbiol., 13 (2022) 1–20. https://doi.org/10.3389/fmicb.2022.1003942.

8. Fischetti V. A. – Lysin therapy for Staphylococcus aureus and other bacterial pathogens. In: Staphylococcus aureus, 409. Springer International Publishing, (2015) 529–540. https://doi.org/10.1007/82_2015_5005.

9. Szafrański S. P., Winkel A., Stiesch M. – The use of bacteriophages to biocontrol oral biofilms. J. Biotechnol., 250 (2017) 29–44. https://doi.org/10.1016/j.jbiotec.2017.01.002.

10. Gilmer D. B., Schmitz J. E., Euler C. W., Fischetti V. A. – Novel bacteriophage lysin with broad lytic activity protects against mixed infection by Streptococcus pyogenes and methicillin-resistant Staphylococcus aureus. Antimicrob. Agents Chemother., 57 (2013) 2743–2750. https://doi.org/10.1128/aac.02526-12.

11. Sharma U., Vipra A., Channabasappa S. – Phage-derived lysins as potential agents for eradicating biofilms and persisters. Drug Discov. Today, 23 (2018) 848–856. https://doi.org/10.1016/j.drudis.2018.01.026.

12. Vázquez R., García E., García P. – Phage lysins for fighting bacterial respiratory infections: A new generation of antimicrobials. Front. Immunol., 9 (2018) 1–12. https://doi.org/10.3389/fimmu.2018.02252.

13. Topka-Bielecka G., Dydecka A., Necel A., Bloch S., Nejman-Faleńczyk B., Węgrzyn G., Węgrzyn A. – Bacteriophage-derived depolymerases against bacterial biofilm. Antibiotics, 10 (2021) 175. https://doi.org/10.3390/antibiotics10020175.

14. Prisinda D., Malinda Y., Lita Y. A., Tjahajawati S. – Diagnosis determination of chronic periapical abscess case using imaging radiography software and microbiological examination in female patients. Padjadjaran J. Dent., 31 (2019) 38–46. https://doi.org/10.24198/pjd.vol31no1.15974.

15. Xie Y., Wahab L., Gill J. – Development and validation of a microtiter plate-based assay for determination of bacteriophage host range and virulence. Viruses, 10 (2018) 189. https://doi.org/10.3390/v10040189.

16. Ács N., Gambino M., Brøndsted L. – Bacteriophage enumeration and detection methods. Front. Microbiol., 11 (2020) 1–7. https://doi.org/10.3389/fmicb.2020.594868.

17. Breed E. S., Murray E. G. D., Smith N. R. – Bergey’s manual of determinative bacteriology, Waverly Press Inc, USA (1957).

18. Kumar S., Stecher G., Li M., Knyaz C., Tamura K. – MEGA X: Molecular evolutionary genetics analysis across computing platforms. Mol. Biol. Evol., 35 (2018) 1547–1549. https://doi.org/10.1093/molbev/msy096.

19. Bala J. D., Lalung J., Al-Gheethi A. A. S., Hossain K., Ismail N. – Microbiota of palm oil mill wastewater in Malaysia. Trop. Life Sci. Res., 29 (2018) 131–163. https://doi.org/10.21315/tlsr2018.29.2.10.