Adaptability to various growth conditions and Anti-Biofilm efficacy of Garlic and Manuka honey on Carbapenem resistant bacteria associated with Urinary Tract Infection

Abstract

Urinary tract infections (UTIs) are a common form of bacterial infections and can be caused by several types of Gram-negative bacteria including Klebsiella pneumonia and Escherichia coli. In recent years, the emergence of carbapenem (the last-resort of antibiotic currently available) resistance among UTIs has challenged healthcare units worldwide and initiated the demand for alternative antimicrobials. It has been reported the hyper resistance activity of these pathogens are closely associated with their adaptability to different growth conditions and their efficacy of Biofilm formation. Bacteria form biofilms that allow their survival in hostile environments. The amount of formed biofilm is affected by external environmental factors. This study investigates the effect of specific parameters such as different growth media, incubation condition, and different growth stage on the amount of biofilm production in Carbapenem resistant Escherichia coli IMP-type) and Klebsiella pneumoniae (OXA-48, NDM-1, and KPC-3). The amount of biofilm formed was measured at different time points, 6, 12, 24 and 48 hours of incubations, different growth conditions, static and shaking and also, investigated the Biofilm formation efficacy of different growth media, nutrient broth, LB broth, and AB broth). Results indicated that there there was a significant difference in biofilm level (p<0.01) when grown under different types of media. Growing under different incubation conditions. Stage of growth of the same species also showed the statistical difference, 30 out of 30 tests (100%) for E. coli and 30 out of 30 tests (100%) for K. pneumoniae. Furthermore, we investigated the effect of known anti-microbials, Garlic and Manuka honey on biofilm formation of these carbapenem resistant strains on different growth media. Our preliminary results showed that both Garlic extract and Manuka honey were effective against Carbapenem sresistant E. coli and K. pneumonia and demonstrated 20-30mg/ml of MIC (Minimum Inhibitory Concentration) for all the test stains. Collectively these findings suggested that biofilm formation is highly affected by incubation conditions, strains’ stage of growth, and media type demonstrating that these conditions may play a role in adaptability of these UTI pathogens on different environmental conditions and their increased prevalence in biofilm associated infections. Additionaly, natural antimicrobials, Garlic and Manuka honey has anti-biofilm efficacy and pave path to identifying alternative anti-microbial therapy on these Carbapenem resistant UTI pathogens.