Parâmetros fisiológicos e moleculares da resposta de Bacteroides fragilis a concentrações subinibitórias de antimicrobianos e avaliação de aspectos da interação bactéria-hospedeiro após infecção experimental

Abstract

Bacteroides fragilis is an anaerobic Gram negative rod member of resident gut microbiota, isolated from clinical infections due in part to its potent virulence factors. As antimicrobial drugs are introduced into the environment, microorganisms respond by becoming resistant and may cause changes in cell physiology. These changes can interfere with the bacterium-host relationship. The aim of this study was to investigate changes in B . fragilis after exposure to ampicillin (AMP), ampicillin/sulbactam (AMS), clindamycin (CLI) and chloramphenicol (CLO); and to investigate changes in host-bacteria relationships after experimental challenge in mice. Four bacterial strains were obtained: BfAMP, BfAMS, BFCLI e BfCLO. Morphological changes, growth patterns, biochemical and physiological abilities, protein expression profile and genetic relationships were comparatively evaluated among the drug-selected strains and Bf parent ATCC 25285. Experimental infection was performed intraperitoneally in conventional mice. Liver and spleen histology, besides cytokines determinations were performed in the intestines. Ampicillin and ampicillin/sulbactam induced the highest significant morphological alterations in the bacterial cells after the initial drug exposure. Decreased sensitivity to all antimicrobials was observed for all drug-selected bacteria strains. Furthermore, drug-exposure was able to induce changes in biochemical pathways and bacterial physiology, if compared with the Bf parent strain. Regarding the ability of biofilm formation, alterations in adherence properties were observed for all drug-selected bacteria, being this characteristic down regulated. The genetic identities were confirmed for the selected strains and no DNA polymorphisms were detected. The cytokine patterns in the intestines from mice challenged with drug-selected bacteria were highly altered, mainly considering the BfAMS strain, as well as the histological aspect concerning the liver and the spleen excised from the experimentally challenged animals. Our results may suggest phenomena related to the gene expression regulation, as a microbial momentary or evolutionary response to the selective pressure of the antimicrobial drugs. These results may also suggest that these drugs have different interactions with the bacterial cell which can be harmful to the host-bacteria relationships. In this way, the data ask for attention regarding the risks of inappropriate chemotherapy, even with low concentrations of the antimicrobial drugs, which might the bacteria genetic expression levels and modulate their pathogenicity