Characterisation of pathogenic Aeromonas veronii and Aeromonas media isolates from aquaculture system: Virulence, antibiotic susceptibility and host mortality assessment
Virulence characterization of Aeromonas veronii & A. media
Keywords:
Aeromonas veronii, A. media, pathogenicity, virulence, antibiotic susceptibilityAbstract
This study presents the identification, virulence characterisation and antibiotic susceptibility of Aeromonas veronii COFCAU_AV1 and Aeromonas media COFCAU_AM, isolated from a diseased Clarius batrachus and aquaculture pond water. Strain identification employed physiological, biochemical and 16S rRNA gene sequence analyses. Both strains demonstrated hemolytic and slime-forming activities and produced DNase, amylase, lipase, gelatinase and caseinase. The LD50 doses in Labeo rohita fingerlings were 106.8 and 107 cells fish-1 for A. veronii and A. media, respectively. A. veronii harboured virulence genes aerA, act, alt, last, exsA, ascC and lip, while A. media had aerA and lip. A. veronii exhibited high resistance to penicillin, erythromycin, vancomycin and nalidixic acid, but high sensitivity to streptomycin, kanamycin and chloramphenicol. A. media showed strong resistance to penicillin and ampicillin but was highly sensitive to chloramphenicol, nalidixic acid and ciprofloxacin. Both strains demonstrated potential to cause mortality and morbidity in L. rohita, with A. veronii exhibiting comparatively higher virulence. This study also reports the pathogenicity of an
A. media strain in Indian major carps for the first time.
Keywords: Aeromonas spp., Antibiotic susceptibility, Clarius batrachus, Labeo rohita, Pathogenicity, Virulence
References
Albert, M. J., Ansaruzzaman, M., Talukder, K. A., Chopra, A. K., Kuhn, I., Rahman, M., ... and Mollby, R. (2000). Prevalence of enterotoxin genes in Aeromonas spp. isolated from children with diarrhea, healthy controls, and the environment. J. Clin. Microbiol., 38(10): 3785-3790. DOI: 10.1128/JCM.38.10.3785-3790.2000
Allen, D. A., Austin, B. and Colwell, R. R. (1983). Aeromonas media, a new species isolated from river water. Int J Syst Evol Microbiol, 33(3): 599-604. DOI: 10.1099/00207713-33-3-599
Austin, B. and Austin, D. A. (2007) Bacterial Fish Pathogens: Diseases of farmed and wild fish. Springer-Praxis, Godalming, UK. DOI: 10.1007/978-3-319-32674-0
Bandeira, J. G. and Baldisserotto, B. (2021) Fish infections associated with the genus Aeromonas: a review of the effects on oxidative status. J. Appl. Microbiol. 131(3): 1083-1101. https://doi.org/10.1111/jam.14986
Bauer, A.W., Kirby, W.M.M., Sherris, J.C. and Turck, M., 1966. Antibiotic susceptibility testing by a standardized single disk method. Am. J. Clin. Pathol., 45: 493-496.
Bondad-Reantaso, M. G., Subasinghe, R. P., Arthur, J. R., Ogawa, K., Chinabut, S., Adlard, R., ... and Shariff, M. (2005). Disease and health management in Asian aquaculture. Vet. Parasitol., 132(3-4): 249-272. DOI: 10.1016/j.vetpar.2005.07.005
Castro-Escarpulli, G., Figueras, M. J., Aguilera-Arreola, G., Soler, L., Fernández-Rendón, E., Aparicio, G. O., ... and Chacon, M. R. (2003). Characterization of Aeromonas spp. isolated from frozen fish intended for human consumption in Mexico. Int. J. Food Microbiol., 84(1): 41-49. DOI: 10.1016/s0168-1605(02)00393-8
Chacón, M. R., Soler, L., Groisman, E. A., Guarro, J., and Figueras, M. J. (2004). Type III secretion system genes in clinical Aeromonas isolates. J. Clin. Microbiol., 42(3): 1285-1287. DOI: 10.1128/JCM.42.3.1285-1287.2004
Chen, F., Sun, J., Han, Z., Yang, X., Xian, J. A., Lv, A., ... and Shi, H. (2019). Isolation, identification and characteristics of Aeromonas veronii from diseased crucian carp (Carassius auratus gibelio). Front. Microbiol., 10: 2742. DOI: 10.3389/fmicb.2019.02742.
Epple, H. J., Mankertz, J., Ignatius, R., Liesenfeld, O., Fromm, M., Zeitz, M., ... and Schulzke, J. D. (2004). Aeromonas hydrophila beta-hemolysin induces active chloride secretion in colon epithelial cells (HT-29/B6). Infect. Immun., 72(8): 4848-4858. DOI: 10.1128/IAI.72.8.4848-4858.2004.
Gaoxue, W., Julin, Y. and Yunkui, Z. (2007). Isolation, identification and drug sensitivity of the pathogens of the skin ulceration disease in Apostichopus japonicus. J. Northwest Agric. For., 35: 87-90.
Gibson, L. F., Woodworth, J. and George, A. M. (1998). Probiotic activity of Aeromonas media on the Pacific oyster, Crassostrea gigas, when challenged with Vibrio tubiashii. Aquaculture, 169(1-2): 111-120. DOI: 10.1016/S0044-8486(98)00369-X.
Gu, J., Fan, Y. and Mitchell, R. (2001). Biological control of zebra mussels by indigenous pathogenic bacteria and their extracellular products. Chinese J Appl Environ Biol, 7: 572-576. DOI: hdl.handle.net/10722/73288.
Han, H. J., Taki, T., Kondo, H., Hirono, I. and Aoki, T. (2008). Pathogenic potential of a collagenase gene from Aeromonas veronii. Can. J. Microbiol., 54(1): 1-10.
Heuzenroeder, M. W., Wong, C. Y. and Flower, R. L. (1999). Distribution of two hemolytic toxin genes in clinical and environmental isolates of Aeromonas spp.: correlation with virulence in a suckling mouse model. FEMS Microbiol. Lett., 174(1): 131-136. DOI: 10.1111/j.1574-6968.1999.tb13559.x.
Hoai, T. D., Trang, T. T., Van Tuyen, N., Giang, N. T. H. and Van Van, K. (2019). Aeromonas veronii caused disease and mortality in channel catfish in Vietnam. Aquaculture, 513: 734425. DOI: 10.1016/j.aquaculture.2019.734425.
Hoel, S., Vadstein, O. and Jakobsen, A. N. (2017). Species distribution and prevalence of putative virulence factors in mesophilic Aeromonas spp. isolated from fresh retail sushi. Front. Microbiol., 8: 931. DOI: 10.3389/fmicb.2017.00931.
Huang, W., Wang, L., Ji, G., Wei, X., Wang, H., Zhao, J., ... and Long, Z. (2013). Identification and drug sensitivity of bacteria Aeromonas veronii and A. media isolated from Chinese sucker Myxocyprinus asiaticus. Fish Sci. (Dalian), 32(4): 210-214.
Janda J. M. and Abbott S. L. (2010) The genus Aeromonas: taxonomy, pathogenicity, and infection. Clin Microbiol, 23: 35-73. DOI: 10.1128/cmr.00039-09.
Karunasagar, I., Karunasagar, I. and Otta, S. K. (2003). Disease problems affecting fish in tropical environments. J. Appl. Aquac., 13(3-4): 231-249. DOI: 10.1300/J028v13n03_03.
Kingombe, C. I. B., Huys, G., Tonolla, M., Albert, M. J., Swings, J., Peduzzi, R. and Jemmi, T. (1999). PCR detection, characterization, and distribution of virulence genes in Aeromonas spp. Appl. Environ. Microbi., 65(12): 5293-5302. DOI: 10.1128/AEM.65.12.5293-5302.1999.
Kirov, S. M., Tassell, B. C., Semmler, A. B., O’Donovan, L. A., Rabaan, A. A. and Shaw, J. G. (2002). Lateral flagella and swarming motility in Aeromonas species. J. Bacteriol., 184(2): 547-555. DOI: 10.1128/JB.184.2.547-555.2002.
Lim, J. and Hong, S. (2020). Characterization of Aeromonas salmonicida and A. sobria isolated from cultured salmonid fish in Korea and development of a vaccine against furunculosis. J. Fish Dis., 43(5): 609-620. DOI: 10.1111/jfd.13158.
Lü, A. J., Hu, X. C., Li, L., Sun, J. F., Song, Y. J., Pei, C., ... and Kong, X. H. (2016b). Isolation, identification and antimicrobial susceptibility of pathogenic Aeromonas media isolated from diseased Koi carp (Cyprinus carpio koi). Iran J Fish Sci. 15: 760-774. DOI: hdl.handle.net/1834/12048.
Lü, A., Song, Y., Hu, X., Sun, J., Li, L., Pei, C., ... and Nie, G. (2016a). Aeromonas veronii, associated with skin ulcerative syndrome, isolated from the goldfish (Carassius auratus) in China. Isr J Aquacult-Bamid 68: 13-21. DOI: hdl.handle.net/10524/54982.
Martin-Carnahan, A. and Joseph, S. W. (2005) Order XII. Aeromonadales ord. nov. In Bergey’s Manual of Systematic Bacteriology. In: Brenner, D. J., Krieg, N. R., Staley, J. T., Garrity, G. M. (eds) Williams & Wilkins: Philadelphia, PA USA.
Mittal, K. R., Lalonde, G., Leblanc, D., Olivier, G. and Lallier, R. (1980). Aeromonas hydrophila in rainbow trout: relation between virulence and surface characteristics. Can. J. Microbiol., 26(12): 1501-1503. DOI: 10.1139/m80-248.
Ørmen, Ø. and Østensvik, Ø. (2001). The occurrence of aerolysin‐positive Aeromonas spp. and their cytotoxicity in Norwegian water sources. J. Appl. Microbiol., 90(5): 797-802. DOI: 10.1046/j.1365-2672.2001.01309.x.
Pradhan, S. K., Devi, R., Khan, M. I. R., Kamilya, D., Choudhury, T. G. and Parhi, J. (2023) Isolation of Aeromonas salmonicida subspecies salmonicida from aquaculture environment in India: Polyphasic identification, virulence characterization, and antibiotic susceptibility. Microb. Pathog. 179: 106100. https://doi.org/10.1016/j.micpath.2023.106100
Reed, L. J. and Muench, H. (1938). A simple method of estimating fifty per cent endpoints. Am. J. Epidemiol., 27(3), 493-497. DOI: 10.1093/oxfordjournals.aje.a118408.
Santos, J. A., González, C. J., Otero, A. and García-López, M. L. (1999). Hemolytic activity and siderophore production in different Aeromonas species isolated from fish. Appl. Environ. Microbi., 65(12): 5612-5614. DOI: 10.1128/AEM.65.12.5612-5614.1999.
Sen, K. and Rodgers, M. (2004). Distribution of six virulence factors in Aeromonas species isolated from US drinking water utilities: a PCR identification. J. Appl. Microbiol., 97(5): 1077-1086. DOI: 10.1111/j.1365-2672.2004.02398.x.
Sha, J., Pillai, L., Fadl, A. A., Galindo, C. L., Erova, T. E. and Chopra, A. K. (2005). The type III secretion system and cytotoxic enterotoxin alter the virulence of Aeromonas hydrophila. Infect. Immun., 73(10): 6446-6457. DOI: 10.1128/IAI.73.10.6446-6457.2005.
Shrestha, M., Xiao, Y., Robinson, H. and Schubot, F. D. (2015). Structural analysis of the regulatory domain of ExsA, a key transcriptional regulator of the type three secretion system in Pseudomonas aeruginosa. PLoS One, 10(8): e0136533. DOI: 10.1371/journal.pone.0136533.
Silver, A. C., Williams, D., Faucher, J., Horneman, A. J., Gogarten, J. P. and Graf, J. (2011). Complex evolutionary history of the Aeromonas veronii group revealed by host interaction and DNA sequence data. PloS one, 6(2): e16751. DOI: 10.1371/journal.pone.0016751.
Smith, P. (2006). Breakpoints for disc diffusion susceptibility testing of bacteria associated with fish diseases: a review of current practice. Aquaculture, 261(4): 1113-1121. DOI: 10.1016/j.aquaculture.2006.05.027.
Sreedharan, K., Philip, R. and Singh, I. S. B. (2013). Characterization and virulence potential of phenotypically diverse Aeromonas veronii isolates recovered from moribund freshwater ornamental fishes of Kerala, India. Antonie Van Leeuwenhoek, 103: 53-67. DOI: 10.1007/s10482-012-9786-z.
Tenover, F. C., Lancaster, M. V., Hill, B. C., Steward, C. D., Stocker, S. A., Hancock, G. A., ... and Hiramatsu, K. (1998). Characterization of staphylococci with reduced susceptibilities to vancomycin and other glycopeptides. J. Clin. Microbiol., 36(4): 1020-1027. DOI: 10.1128/JCM.36.4.1020-1027.1998.
Tian, Z., Cheng, S., Xia, B., Jin, Y., Bai, F., Cheng, Z., ... and Wu, W. (2019). Pseudomonas aeruginosa ExsA regulates a metalloprotease, ImpA, that inhibits phagocytosis of macrophages. Infect. Immun., 87(12): 10-1128. DOI: 10.1128/IAI.00695-19.
Torlak, E., Korkut, E., Uncu, A. T., and Şener, Y. (2017). Biofilm formation by Staphylococcus aureus isolates from a dental clinic in Konya, Turkey. J. Infect. Public Healt., 10(6): 809-813. DOI: 10.1016/j.jiph.2017.01.004.
Vilches, S., Urgell, C., Merino, S., Chacón, M. R., Soler, L., Castro-Escarpulli, G., ... and Tomás, J. M. (2004). Complete type III secretion system of a mesophilic Aeromonas hydrophila strain. Appl. Environ. Microbi., 70(11): 6914-6919. DOI: 10.1128/AEM.70.11.6914-6919.2004.
Yao, D., Bing, X., Zhu, M., Bi, K., Chen, L. and Zhang, X. (2010). Molecular identification and drug resistance of pathogenic Aeromonas sobria isolated from Misgurnus anguillicaudatus. Oceanologia et Limnologia Sinica/Hai Yang Yu Hu Chao, 41(5): 756-762.
Yesmin, S., Rahman, M. H., Hussain, M. A., Khan, A. R., Pervin, F. and Hossain, M. A. (2004). Aeromonas hydrophila infection in fish of swamps in Bangladesh. Pak. J. Biol. Sci. 7: 409-411. DOI: 10.3923/pjbs.2004.409.411
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