Investigation of genes responsible for tetracycline, macrolide, sulfamethoxazole and trimethoprim resistance in bacteria detected from water samples collected from the environment

Water is one of the most important bacterial habitats in the world. It is also one of the main ways of spreading bacteria in different environments. Resistance of bacteria to environmental conditions such as water and their ability to multiply in this environment is the source of various gene transfers between bacteria. This study was conducted to investigate whether bacteria isolated from 7 environmental water samples (snow and river water) in Warsaw have plasmid-mediated antibiotic resistance genes, and whether these genes can be transferred if they carry these genes. Primer sequences specific for the gene responsible for tetracycline, macrolide, sulfamethoxazole and trimethoprim resistance (tet A, tet W, tet O, tet Q and tet C, erm A, erm B, ermC, sul1 gene, DHFR I gene) were used in water samples. In addition, the target region of the gene responsible for lateral gene transfer (conjugation) was investigated. Gene compartments were investigated by real-time PCR method at Warsaw Medical University. Identification of species and genera was made by using conventional methods and Vitek2 device (bioMeriux, France) at Gaziantep University. DNA sequence analysis (Sanger) was performed to confirm strains with resistance gene. Resistance transfer was performed to the non-resistant E.coli strains by electroporation from samples containing plasmid or gene transfer elements. While none of the resistance genes tested in the Susic river were detected, all resistance gene regions tested in the Vistula river were detected. All gene regions except snow erm C were detected in the snow sample. The most common gene in rivers is tet C and it was detected in 5 different rivers. After sequence analysis of vistula and snow water samples, conjugation facilitating ob mob 'proteins were found in Vistula River and 2 plasmids were found in snow water sample. Electroporation of these two specimens carrying transfer protein showed resistance resistance in non-resistant E.coli strains by resistance gene transfer. Our research showed that resistant bacteria found in the environment could create gene reservoirs among other bacteria.