ANALYZE OF RESISTANCE-RELATED GENES USING SOFTWARE RESFINDER AND AGAR DIFFUSION TECHNIQUE OF THE FIRST GENOME SEQUENCED Proteus mirabilis ISOLATED FROM TRACHEAL SECRETION IN BRAZIL

Proteus spp. bacteria are mostly known as opportunistic human pathogens, causes different nosocomial infections. Among the species of this genus, the most prevalent in infections in humans is Proteus mirabilis, a bacilli Gram-negative bacterium of Enterobacteriaceae family. The P. LBUEL-H11 strain was isolated in 2015 from tracheal secretion of a hospitalized inpatient admitted to the University Hospital of the State University of Londrina in South of Brazil. This is the first P. mirabilis multidrug-resistant isolated from tracheal secretion that have the whole genome sequenced (GenBank acession QGGA01000000). We observed important resistance determinant genes in an estimated genome size of 4,534,929 bp. P. mirabilis carry multiple antibiotic resistance genes, including strains Extended-spectrum beta-lactamase (ESBL) producing, representing a risk for the treatment of different forms of infections. Its clinical significance is that ESBL bacteria express β-lactamase enzyme that hydrolyze most β-lactam antimicrobials, including cephalosporins, cephamycins, penicillins, limiting the therapeutic options for treatment of infections caused by these bacteria. The development of antimicrobial resistance among Gram-negative pathogens has been progressive. The objective of this study was to phenotypically and in silico evaluate the antimicrobial susceptibility profile of the sequenced P. mirabilis strain isolated from tracheal secretion. The antimicrobial susceptibility assay was performed by the agar diffusion technique, using the antimicrobials recommended by the Clinical and Laboratory Standards Institute (CLSI -2015), where we observed that the strain was resistant to all antimicrobials tested (aztreonam, ampicillin, ampicillin-sulbactan, amikacin, cefalotine, cefuroxime, gentamicin, ceftazidime, cefepime, ciprofloxacin, cefetrioxane and trimethoprim-sulfamethoxazole). This isolate was ESBL producer. Resistance-related genes were analyzed in silico using ResFinder 3.0 (acquired antimicrobial resistance gene finder), available from the Center for Genomic Epidemiology website (cge.cbs.dtu.dk/services/ResFinder/). Using this approach, we identified resistance genes to aminoglycosides, aadA2 (Acession JQ364967), aph(3’)-VIa (Acession X07753), aph(3’’)-Ib (Acession AF321551), aph(6)-Id (Acession M28829), aac(3)-IId (Acession EU022314); beta-lactam, blaCTX-M-14 (Acession AF252622), blaTEM-1B (Acession JF910132); phenicol, cat (Acession M11587); sulphonamide, sul1 (Acession CP002151), sul2 (Acession GQ421466); tetracycline, tet(J) (Acession ACLE01000065), tet(G) (Acession AF133140); trimethoprim, dfrA12 (Acession AB571791). Therefore, P. mirabilis LBUEL-H11 is a multidrug resistant bacterium and the tests (Agar diffusion technique and software ResFinder) were completed because the phenotypically results observed was confirmed in the in silico analyse.