Safety characteristics of beneficial vaginal lactic acid bacteria (LAB) to be included in a probiotic product

ANTONELLA MARCHESI; JESSICA SILVA; CECILIA ARISTIMUÑO FICOSECO; MARIA ELENA FÁTIMA NADER

Mar del Plata

Congreso; XLVIII REUNION ANUAL DE LA ASOCIACION ARGENTINA DE FARMACOLOGIA EXPERIMENTAL (SAFE); 2016

SAFE-SAI-SAIC

The urogenital tract infections are the most common cause of attendance to gynecologist consult, being frequently treated with antibiotics. Antibiotic use may contribute to the emergence of antibiotic resistance bacteria in humans, because genes spreading through food chains may occur. Consequently, the addition of probiotics into medical treatments has gained increasing importance in health and constituting a novel alternative to replace the excessive use of antibiotic. Our research group had been investigating lactic acid bacteria isolated from female human vagina, beneficial properties and their advantages which a phytobiotic formulation. In this study, safety characteristics such as antibiotic resistance genes and virulence factors as hyaluronidase, gelatinase and hemolysis of LAB strains intended to be used as animal probiotic formula, were evaluated. The antibiotic resistance had examined by phenotypic and genetic assays using the following antimicrobials: Ampicillin, Tetracycline, Chloramphenicol, Gentamicin, Streptomycin, Kanamycin, Vancomycin and Erythromycin. The minimum inhibitory concentration (MIC) assays been performed by the plate diffusion technique in the agar and wells filled in LSM media, incubated in microaerophylic conditions at 37°C. The evaluation was performed with different lactobacilli species (Lactobacillus reuteri, L. gasseri, L. rhamnosus, L. salivarius) and the European Food Safety Authority (EFSA) proposed cutoff values applied. By PCR using specific primers, the presence of antibiotic resistance genes (vanA, vanB and vanX erm(C), bla, cat, erm(B), aac(6´)-aph(2´´), aph(3´´)-III, strA, strB, aadA, aadE, ant(6), tet(M), tet(K), tet(L), tet(S)) were evaluated, whereas phenotypic assays in agar solid media and well liquid dilution with specific substrates, were used for virulence factors detection. There is a wide degree of coincidence between the results obtained in the phenotypic and genotypic evaluation of the resistance of the strain to the different antibiotic. Furthermore, the results indicates a resistance to ERY (80% of coincidence between phenotypic and genotypic study) for all the strains testing. Although, L. rhamnosus 1508 and L. rhamnosus 1511 showed the presence of coding genes for TET/VAN. In addition, the results indicated L. gasseri 1267 and L. reuteri 1327 were resistance to CLIN (50%) and KAN (50%) in phenotypic assay. Even though, most of the strains were sensibility to CHLOR/STRE/GEN, which are the antibiotic frequently applied to the therapy of urogenital infections. . All of the assayed lactobacillus strains were free of virulence factors. With the results obtained to ICM and virulence factors, showed that some of the strain with beneficial characteristic could be introduced into a pharmabiotic product to restore the vaginal microbiota as an alternative therapy to other pathogens.