Abstract

Objective: To assess the antimicrobial activity of the Actinobacteria bioactive secondary metabolite and characterize the drug resistance mechanisms of Pseudomonas aeruginosa (P. aeruginosa) and Acinetobacter baumannii (A. baumannii). Methods: Potential marine Actinobacteria were isolated and the crude extract was purified using thin layer chromatography, the fractions were tested for antimicrobial activity and phylogeny of the selected strain was analyzed. Isolated pathogenic strains were screened for extended spectrum beta-lactamase, mannan-binding lectin, AmpC production, efflux mechanism and polymerase chain reaction. The cephalosporin and carbapenem antibiotics were synergistically tested along with Streptomyces sp. PM49 fraction by combination disc test and double-disc synergy test. Heterogeneous susceptibility assay, minimum inhibitory concentration and expression of DnaK (Hsp70) were determined. Results: Streptomyces sp. PM49 active fraction of Rfvalue 0.69 showed antimicrobial activity and an inhibitory zone of 15 to 7 mm obtained. About 34.1% of P. aeruginosa and 4.8% of A. baumannii were multiple drug resistant. AmpC β-lactamase was found in 12% of A. baumannii, efflux mechanism was putatively positive in 8/23 of P. aeruginosa and 3/20 of A. baumannii. Combination disc test and double-disc synergy test with both PM49 compound and antibiotics showed an increase in the inhibitory zone of <3 mm to 4 mm, three P. aeruginosa isolates expressed blaIMP. Heteroresistant subcolonies grew at a frequency of 3 ×10-5 to 1 ×10-5. Stress induction analysis showed increase of DnaK during heat shock at 52 °C, the levels of protein doubled after exposure to the antibiotics. Conclusions: Novel unexplored Streptomyces spp. antimicrobial constituents can be developed as an inhibitor and can be substituted along with the available antibiotics to combat the drug resistant pathogens.

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