Isolation and Characterization of Lytic Bacteriophages against Aeromonas dhakensis Isolated from Water in Thailand
Keywords:Aeromonas dhakensis, bacteriophage, biocontrol, lytic bacteriophages
Phage therapy has emerged as a potential solution to the issue of multidrug-resistant bacteria. In this study, a novel bacteriophage vB_AdhS_M4 , which infects Aeromonas dhakensis, was isolated. A. dhakensis strains were isolated from water samples in Thailand and identified by biochemical and 16S rDNA sequence analysis. Four isolates, namely AM, NGP8, AK3, and SBKN4 were identified as A. dhakensis and used as hosts for phage isolation. Only 1 phage, vB_AdhS_M4, was obtained using A. dhakensis AM as host. Phage vB_AdhS_M4 morphology, host range, one-step growth curve, pH and temperature stability and genome were investigated. Transmission electron microscopy revealed that vB_AdhS_M4 has an icosahedral head of 64.8 ± 0.51 nm and a long tail of 185.4 ± 0.48 nm, suggesting that it belongs to the Siphoviridae family. Phage vB_AdhS_M4 had a latent period of 50 mins and a mean burst size of approximately 48 phage particles per infected cell. Our study on host-range determination demonstrated that this phage was specific for infecting A. dhakensis. Phage vB_AdhS_M4 was stable within a pH range of 4-7 and at temperatures ranging from 4 to 45 °C. The complete genome of Aeromonas phage vB_AdhS_M4 is 61,429 bp in length, with a G+C content of 61.7% and 77 open reading frames. In vitro A. dhakensis growth inhibition were observed with vB_AdhS_M4 at various MOIs. The greatest reduction in cell count occurred during the first 6-12 h of incubation with the phage, compared to the uninfected bacterial control, at all MOIs tested. Although the bacteria regrew after 24 h of incubation with the phage, the bacterial count remained lower than that of the control throughout the entire 48-h period. Based on these findings, it appears that vB_AdhS_M4 could be a valuable tool for biocontrol of A. dhakensis in aquaculture.
Adams, M. H. (1959). Bacteriophages. New York: Interscience Publishers.
Akmal M., Rahimi-Midani A., Hafeez-Ur-Rehman M., Hussain A., & Choi T. J. (2020). Isolation, Characterization and application of a bacteriophage infecting the fish pathogen Aeromonas hydrophila. Pathogens, 9, 215.
Bai, M., Cheng, Y. -H., Sun, X. -Q., Wang, Z. -Y., Wang, Y. -X., Cui, X. -L., & Xiao, W. (2019). Nine novel phages from a plateau lake in southwest China: insights into Aeromonas phage diversity. Viruses, 11(7), 615.
Bankevich, A., Nurk, S., Antipov, D., Gurevich, A. A., Dvorkin, M., Kulikov, A. S., Lesin, V. M., Nikolenko, S. I., Pham, S., Prjibelski, A. D., Pyshkin, A. V., Sirotkin, A. V., Vyahhi, N., Tesler, G., Alekseyev, M. A., Pevzner, P. A. (2012). SPAdes: A new genome assembly algorithm and its applications to single-cell sequencing. Journal of Computational Biology, 19(5), 455-477.
Beaz-Hidalgo, R., Martinez-Murcia, A., & Figueras, M. J. (2013). Reclassification of Aeromonas hydrophila subsp. dhakensis Huys et al., 2002 and Aeromonas aquariorum Martinez-Murcia et al., 2008 as Aeromonas dhakensis sp. nov. comb nov. and emendation of the species Aeromonas hydrophila. Systematic and Applied Microbiology, 36(3), 171–176.
Bolger, A. M., Lohse, M., & Usadel, B. (2014). Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics, 30, 2114-2120.
Chan, B. K., Abedon, S. T., & Loc-Carrillo, C. (2013). Phage cocktails and the future of phage therapy. Future Microbiology, 8, 769–783.
Chan, P. P., & Lowe, T. M. (2019). tRNAscan-SE: searching for tRNA genes in genomic sequences. Gene prediction: methods and protocols, 1-14.
Chen, P. L., Lamy, B., & Ko, W. C. (2016). Aeromonas dhakensis, an Increasingly Recognized Human Pathogen. Frontiers in Microbiology, 7, 793
Cheng, Y., Gao, D., Xia, Y., Wang, Z., Bai, M., Luo, K., Cui, X., Wang, Y., Zhang, S., & Xiao, W. (2021). Characterization of novel bacteriophage AhyVDH1 and its lytic activity against Aeromonas hydrophila. Current Microbiology, 78(1), 329-337.
Ding, T., Sun, H., Pan, Q., Zhao, F., Zhang, Z., & Ren, H. (2020). Isolation and characterization of Vibrio parahaemolyticus bacteriophage vB_VpaS_PG07. Virus Research, 286, 198080.
Esteve, C., Alcaide, E., & Blasco, M. D. (2012). Aeromonas hydrophila subsp. dhakensis isolated from feces, water and fish in Mediterranean Spain. Microbes and Environments, 27, 367–373.
Figueras, M. J., Alperi, A., Saavedra, M. J., Ko, W. C., Gonzalo, N., Navarro, M., & Martinez-Murcia, A. J. (2009). Clinical relevance of the recently described species Aeromonas aquariorum. Journal of Clinical Microbiology, 47(11), 3742–3746.
Grant, J. R., & Stothard, P. (2008). The CGView Server: a comparative genomics tool for circular genomes. Nucleic Acids Research, 36(2), W181-W184.
Gurney, J., Aldakak, L., Betts, A., Gougat-Barbera, C., Poisot, T., Kaltz, O., & Hochberg, M. E. (2017). Network structure and local adaptation in co-evolving bacteria-phage interactions. Molecular Ecology, 26, 1764–1777.
Huys, G., Kampfer, P., Albert, M.J., Kuhn, I., Denys, R., & Swings, J. (2002). Aeromonas hydrophila subsp. dhakensis subsp. nov., isolated from children with diarrhoea in Bangladesh, and extended description of Aeromonas hydrophila subsp. hydrophila (Chester 1901) Stanier 1943 (approved lists 1980). International Journal of Systematic and Evolutionary Microbiology, 52(3), 705–712.
Kabwe, M., Brown, T., Speirs, L., Ku, H., Leach, M., Chan, H.T., & Tucci, J. (2020). Novel bacteriophages capable of disrupting biofilms from clinical strains of Aeromonas hydrophila. Frontiers in Microbiology, 11, Article 194.
Kühn, I., Albert, M. J., Ansaruzzaman, M., Bhuiyan, N. A., Alabi, S. A., Islam, M. S., Neogi, P. K., Huys, G., Janssen, P., Kersters, K., & Möllby, R. (1997). Characterization of Aeromonas spp. isolated from humans with diarrea, from healthy controls, and from surface water in Bangladesh. Journal of Clinical Microbiology, 35, 369–373.
Lane, D. J. (1991). 16S/23S rRNA sequencing. New York: Wiley.
Martin-Carnahan, A., & Joseph, S. W. (2005). Order XII. Aeromonadales ord. nov., in Brenner, D.J., Krieg, N.R., Staley, J.T., Garrity G.M. (Eds), Bergey’s Manual of Systematic Bacteriology, Philadelphia, PA, USA: Williams & Wilkins.
Pringsulaka, O., Patarasinpaiboon, N., Suwannasai, N., Atthakor, W., & Rangsiruji, A. (2011). Isolation and characterisation of a novel Podoviridae-phage infecting Weissella cibaria N 22 from Nham, a Thai fermented pork sausage. Food Microbiology, 28(3), 518-525.
Pu, W., Guo, G., Yang, N., Li, Q., Yin, F., Wang, P., Zheng, J., & Zheng, J., (2019). Three species of Aeromonas (A. dhakensis, A. hydrophila and A. jandaei) isolated from freshwater crocodiles (Crocodylus siamensis) with pneumonia and septicemia. Letters in Applied Microbiology, 68(3), 212–218.
Sambrook, J., Fritsch, E. R., & Maniatis, T. (1989). Molecular cloning: a laboratory manual. 2nd ed. New York: Cold Spring Harbor.
Seemann, T. (2014). Prokka: rapid prokaryotic genome annotation. Bioinformatics, 30, 2068-2069.
Sinha S, Shimada T, Ramamurthy T, Bhattacharya SK, Yamasaki S, Takeda Y, & Nair GB. (2004). Prevalence, serotype distribution, antibiotic susceptibility and genetic profiles of mesophilic Aeromonas species isolated from hospitalized diarrhoeal cases in Kolkata, India. Journal of Medical Microbiology, 53, 527–534.
Sunthornthummas, S., Doi, K., Rangsiruji, A., Sarawaneeyaruk, S., & Pringsulaka, O. (2017). Isolation and characterization of Lactobacillus paracasei LPC and phage ΦT25 from fermented milk. Food Control, 73(8), 1353-1361.
Wright, R. C. T., Friman, V. -P., Smith, M. C. M., & Brockhurst, M. A. (2018). Cross-resistance is modular in bacteria–phage interactions. PLoS Biology, 16, e2006057.
Yuan, S., Chen, L., Liu, Q., Zhou, Y., Yang, J., Deng, D., Li, H., & Ma, Y. (2018). Characterization and genomic analyses of Aeromonas hydrophila phages AhSzq-1 and AhSzw-1, isolates representing new species within the t5virus genus. Archives of Virology, 163, 1985–1988.
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