Muhammed DUMAN, Izzet Burcin SATİCİOGLU, Burak OZDEMİR, Nihed AJMİ, Soner ALTUN

Determination of Opportunistic Pathogens and Antimicrobial Resistance Characterization Isolated From Rainbow Trout in Turkey

Modern aquaculture enables effective means for intensive aquaculture production under “controllable” conditions. This rapidly growing industry, however, has experienced relatively severe disease problems owing to a lack of control of the microbiota in rearing systems. In the present study, we identified the opportunistic pathogens and some members of natural microbiota, which were obtained from our culture collection by culturable methods and characterized biochemically in addition to molecular analysis. All isolates were identified by gene sequence analysis and confirmed taxonomically in the bacterio.net database. The isolates were characterized by means of antimicrobial susceptibilities by broth microdilution method analysis, and the resistance gene determinants were screened by PCR analysis. A total of 14 species were identified with high genetic similarity in the GenBank database. MIC results showed that bacteria have heterogeneous characteristics for the susceptibility of an agent into the genus, and species have high MIC values for sulfamethoxazole, trimethoprim, and ampicillin comparing to other agents. A total of 13 different resistance genes were determined in the bacteria, and some of them have multiple resistance genes up to five genes.

Keywords:

Opportunistic pathogens, Microbial pathogenesis, Antimicrobial resistance genes, Natural Microbiota,

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1. Austin B, Austin DA, Austin B, Austin DA. Miscellaneous Pathogens. In: Austin B, Austin DA, eds. Bacterial Fish Pathogens. Springer International Publishing; 2016:603-642. doi:10.1007/978-3-319-32674-0_11
2. Öztürk RÇ, Altinok I. Bacterial and Viral Fish Diseases in Turkey. Turkish J Fish Aquat Sci. 2014;14(1):275-297. doi:10.4194/1303-2712-v14_1_30
3. Duman M, Saticioglu IB, Buyukekiz AG, Balta F, Altun S. Molecular characterization and antimicrobial resistance profile of atypical Citrobacter gillenii and Citrobacter sp. isolated from diseased rainbow trout (Oncorhynchus mykiss). J Glob Antimicrob Resist. 2017;10:136-142. doi:10.1016/j.jgar.2017.05.014
4. Duman M, Buyukekiz AG, Saticioglu IB, Cengiz M, Sahinturk P, Altun S. Epidemiology, genotypic diversity, and antimicrobial resistance of Lactococcus garvieae in farmed rainbow trout (Oncorhynchus mykiss. Iran J Fish Sci. 2020;19(1):1-18. doi:10.22092/ijfs.2018.117609
5. Saticioglu IB, Duman M, Wiklund T, Altun S. Serological and genetic characterization of Flavobacterium psychrophilum isolated from farmed salmonids in Turkey. J Fish Dis. 2018;41(12):1899-1908. doi:10.1111/jfd.12901
6. Austin B, Newaj-Fyzul A, eds. Diagnosis and Control of Diseases of Fish and Shellfish. Chichester, UK: John Wiley & Sons, Ltd; 2017. doi:10.1002/9781119152125
7. Derome N, Boutin S, Llewellyn M, Gauthier J. The Rasputin effect: when commensals and symbionts become parasitic. 2016.
8. Svetlana J, Dobrila JD, Veljović LJ. Citrobacter freundii as a cause of disease in fish. Acta Vet Brno. 2003;53(5-6):399-410. doi:10.2298/AVB0306399J
9. Paździor E. Shewanella putrefaciens - A new opportunistic pathogen of freshwater fish. J Vet Res. 2016;60(4):429-434. doi:10.1515/jvetres-2016-0064
10. Fink C, Roeder T. How Well Do Surrogate Hosts Serve as Model Systems for Understanding Pathogenicity. In: Springer, Cham; 2016:3-25. doi:10.1007/978-3-319-28170-4_1
11. Sørensen SJ, Bailey M, Hansen LH, Kroer N, Wuertz S. Studying plasmid horizontal transfer in situ: a critical review. Nat Rev Microbiol. 2005;3(9):700-710. doi:10.1038/nrmicro1232
12. Cabello FC, Godfrey HP, Tomova A, et al. Antimicrobial use in aquaculture re-examined: its relevance to antimicrobial resistance and to animal and human health. Environ Microbiol. 2013;15(7):1917-1942. doi:10.1111/1462-2920.12134
13. Marchesi JR, Sato T, Weightman AJ, et al. Design and evaluation of useful bacterium-specific PCR primers that amplify genes coding for bacterial 16S rRNA. Appl Environ Microbiol. 1998;64(2):795-799. doi:10.1128/aem.64.2.795-799.1998
14. Mulet M, Bennasar A, Lalucat J, García-Valdés E. An rpoD-based PCR procedure for the identification of Pseudomonas species and for their detection in environmental samples. Mol Cell Probes. 2009;23(3-4):140-147. doi:10.1016/j.mcp.2009.02.001
15. Duman M, Saticioglu IB, Altun S. Molecular characterization and antimicrobial resistance profile of fecal contaminants and spoilage bacteria that emerge in rainbow trout (Oncorhynchus mykiss) farms. Biosci Microbiota, Food Heal. 2019;38(2):41-48. doi:10.12938/bmfh.18-007
16. CLSI, Institute Clinical and Laboratory Standards. Methods for Broth Dilution Susceptibility Testing of Bacteria Isolated From Aquatic Animals; Proposed Guideline. In: CLSI Document M49-P. Vol 25. ; 2006:47. www.clsi.org.
17. CLSI - Clinical and Laboratory Standards Institute, 2014 undefined. Performance standards for antimicrobial susceptibility testing of bacteria isolated from aquatic animals; second informational supplement.
18. Duman M, Altun S, Cengiz M, Saticioglu IB, Buyukekiz AG, Sahinturk P. Genotyping and antimicrobial resistance genes of Yersinia ruckeri isolates from rainbow trout farms. Dis Aquat Organ. 2017;125(1):31-44. doi:10.3354/dao03132
19. Saticioglu IB, Duman M, Altun S. Antimicrobial resistance and molecular characterization of Pantoea agglomerans isolated from rainbow trout (Oncorhynchus mykiss) fry. Microb Pathog. 2018;119:131-136. doi:10.1016/j.micpath.2018.04.022
20. Austin B, Austin DA. Enterobacteriaceae Representatives. In: Bacterial Fish Pathogens. Cham: Springer International Publishing; 2016:323-396. doi:10.1007/978-3-319-32674-0_6
21. Olafsen JA. Interactions between fish larvae and bacteria in marine aquaculture. In: Aquaculture. Vol 200. Elsevier; 2001:223-247. doi:10.1016/S0044-8486(01)00702-5
22. Saticioglu IB, Duman M, Altun S. Antimicrobial resistance and molecular characterization of Pantoea agglomerans isolated from rainbow trout (Oncorhynchus mykiss) fry. Microb Pathog. 2018;119. doi:10.1016/j.micpath.2018.04.022
23. Lehane L, Rawlln GT. Topically acquired bacterial zoonoses from fish: a review. Med J Aust. 2000;173(5):256-259. doi:10.5694/j.1326-5377.2000.tb125632.x
24. Chacko, K L, Rajagopal, V, Jaibi, K, Latha, C, Nanu E. Fish borne bacterial zoonoses. Intas Polivet. 2006;7(11):207-211. https://www.cabdirect.org/cabdirect/abstract/20073017434. Accessed April 22, 2020.
25. Lowry T, Smith SA. Aquatic zoonoses associated with food, bait, ornamental, and tropical fish. J Am Vet Med Assoc. 2007;231(6):876-880. doi:10.2460/javma.231.6.876
26. International NT-MA, 2011 undefined. Evaluating the importance of zoonotic bacteria, antimicrobial use and resistance in aquaculture and seafood.
27. Smith P. Antimicrobial Resistance in Aquaculture. Vol 27.; 2008.
28. Weir M, Rajić A, Dutil L, Uhland C, Bruneau N. Zoonotic bacteria and antimicrobial resistance in aquaculture: Opportunities for surveillance in Canada. Can Vet J. 2012;53(6):619-622.