Determination of plasmit-delivered colistine resistance genes in trout in Tunceli by multiplex polymeraze chain reaction

Determination of Plasmit-Delivered Colistine Resistance Genes in Trout in Tunceli by Multiplex Polymeraze Chain Reaction Antimicrobial resistance (AMR) is a global health problem. Aquatic ecosystems can provide an ideal environment for the spread of AMR. Antimicrobials added to the feed as therapeutic agents in fish farms are added into the water. Antibiotic residues remain in aquatic environments, and these wastewaters can be mixed with rivers and transported to production facilities, and resistance genes can be transferred to bacteria in that environment. According to the researches, there is not enough study on the determination of the antimicrobial resistance of fishery products in Turkey, but the studies are limited. In this study, bacteria isolation is it was aimed to determine by conventional methods from 64 water samples and 600 samples taken from the intestine, skin and gill parts of 200 juvenile or adult rainbow trout obtained from 16 facilities where rainbow trout breeding in net cages and concrete ponds in Pertek, Çemişgezek, Ovacık, Mazgirt districts of Tunceli province during the months of June, July, and December. isolation from 600 samples taken from the intestine, skin and gill parts of 200 juvenile or adult rainbow trout and 64 water samples obtained from 16 facilities where rainbow trout are raised in net cages and concrete ponds in Pertek, Çemişgezek, Ovacık, Mazgirt districts of Tunceli province during the months of June, July and December, to identification of bacteria by Matrix Mediated Laser Desorption Ionization-Time of Flight-Mass Spectrometer (MALDI-TOF MS) analysis, To reveal the presence of mcr gene by multiplex PCR method using DNA extracted from bacteria species belonging to the identified enterobacteria family, to determine the colistin minimum inhibitory (MIC) concentration, liquid microdilution and antimicrobial susceptibility of bacterial isolates belonging to this family by Kirby-Bauer disk diffusion methods. In this study; 290 (59.67%) of the mesh cages belonging to 8 bacterial families, including Moraxellaceae, Enterococcaceae, Vibrionaceae, Comamonadaceae, Pseudomonas, Staphylococcaceae, Pseudomonadaceae, Enterobacteriaceae and 96 (40.33%) of concrete pools, a total of 486 bacterial identification was performed by MALDI-TOF MS analysis. Bacterial growth was not observed in 114 samples. The mcr gene could not be detected in bacteria belonging to the enterobacteria family by multiplex PCR method. Colistin susceptibility was revealed by the MIC concentration being less than 2 as a result of the liquid microdilution test. As a result of the antimicrobial sensitivities determined by the Kirby-Bauer disc diffusion method, it was determined that the isolates belonging to the enterobacteria family developed resistance to the quinolone group antibiotics.The first attempt to determine the presence of colistin-resistance genes in enterobacterales containing E.coli, Enterobacter and Salmonella species in our country and in the Eastern Anatolia basin is to reveal the original aspect of the project. When it compared with conventional and molecular identification methods, it is more effective on per sample costs and elapsed time on working. In particular, MALDI-TOF MS analysis, which can achieve 100% results in typing bacteria belonging to the Enterobacteria family, has come to the fore in molecular studies. In our study, which covers rainbow trout farms in Mazgirt, Çemişgezek, Ovacık and Pertek districts of Tunceli province, we will contribute to our country's antimicrobial resistance prevention strategies by revealing the resistance potential and danger of plasmid-mediated colistin. Keywords: Antimicrobial resistance, Escherichia coli, Colistin, Multiplex PCR, MALDI-TOF MS Analysis.