Determination of genes coding for antibiotic and copper resistance in Enterococcus bacteria

U ovom radu analizirani su izolati enterokoka iz fecesa divlje svinje (n=30), spontano fermentiranih kobasica proizvedenih od mesa divlje i domaće svinje (n=10) i Istarskog sira (n=5). Multipleks PCR metodom utvrđeni su geni koji kodiraju za rezistenciju na antibiotike: eritromicin (ermB), tetraciklin (tetM), gentamicin (aac6-aph2), kanamicin (aphA3), vankomicin (vanC1, vanA, vanB, vanC2) te gen za rezistenciju na bakar (tcrB). Gen ermB je detektiran kod 33,33 % izolata iz fecesa, što je najviše od analiziranih ekosustava. Izolati fecesa sadrže 80 % gena tetM, izolata iz kobasice sadrže 20 % i niti jedan izolat iz sira. Najveći postotak gena za kanamicin (100 %) utvrđen je kod enterokoka prikupljeni iz sira, dok 90 % izolata fecesa te 60 % iz kobasice sadrži gen za rezistentnost na isti antibiotik. Detektirani su geni vanC1, vanA, vanB, vanC2 kod 90 % izolata iz kobasice, dok iz fecesa i sira navedene gene sadrži 80 % izolata. Kod analiziranih izolata nije bilo značajne korelacije između detektiranih gena tcrB i aac6-aph2. Naši rezultati upućuju da se u prikupljenim izolatima enterokoka pojavljuju geni koji kodiraju za rezistenciju na antibiotike i bakar bez obzira na ekosustav, a trend posjedovanja gena za rezistentnost je sličan.In this paper, fecal enterococci isolates from wild boar pigs (n = 30), spontaneously fermented sausages produced from wild boar and homemade pigs (n = 10) and Istrian cheese (n = 5) were analyzed. Multiplex PCR methods have been used to determine the genes that encode for resistance to antibiotics: erythromycin (ermB), tetracycline (tetM), gentamicin (aac6-aph2), kanamycin (aphA3), vancomycin (vanC1, vanA, vanB, vanC2) and the copper resistance gene (tcrB). Gen ermB was detected in 33.33% of the isolates from the feces, which is the most of the analyzed ecosystems. Feces isolates contain 80% tetM genes, 20% sausage isolates contain that same gene and no cheese isolates contain it. The highest percentage of kanamycin gene (100%) was found in enterococci collected from the cheese, while 90% of fecal isolates and 60% of sausage isolates showed resistance to the same antibiotic. Genes vanC1, vanA, vanB, vanC2 are detected in 90% of sausage isolates, while from feces and cheese said gene contains 80% of isolates. There was no significant correlation between the presence of genes tcrB and aac6-aph2 in the analyzed isolates. Our findings point out that genes coding for resistance to antibiotics and copper, regardless of the ecosystem, appear in the collected gastrointestinal isolates, and the gene resistance trend is similar