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Microbial biofilm are factions of surface-colonized cells encompassed in a matrix of self-secreted extracellular polymeric secretions. Profound application of antibiotics inorder to treat infections due to microbial biofilms has led to the emergence of several strains that exhibit it’s resistance to the drugs. A novel type of 3,6-di(pyridin-2-yl)-1,2,4,5-tetrazine (pytz) capped silver nanoparticle (TzAgNPs) was synthesized and efforts were given to test it’s activity against microbes and biofilms, Pseudomonas aeruginosa in this regard, a widely used biofilm forming pathogenic organism. The synthesized TzAgNPs showed considerable antimicrobial activity wherein the MIC value of TzAgNPs was found at 40 μg/ml against Pseudomonas aeruginosa. Antibiofilm activity of TzAgNPs was tested against Pseudomonas aeruginosa by carrying out an array of experiments like microscopic observation, crystal violet assay and protein count using the sub-MIC concentrations of TzAgNPs. Since TzAgNPs showed efficient antibiofilm activity, thus, in the current study, we investigated the underlying cause of biofilm attenuation of Pseudomonas aeruginosa using TzAgNPs. To this end, we discerned that the sub-MIC doses of TzAgNPs increase the ROS level considerably in the bacterial cell. The result showed that the ROS level and microbial biofilm formation are inversely proportional. Thus, the attenuation in microbial biofilm could be attributed to the accumulation of ROS level. Furthermore, it was also duly noted that microorganisms upon treatment with TzAgNPs exhibited considerable diminution in virulence factors (Protease and pyocyanin) contrasted to control where the organisms were untreated with TzAgNPs. Thus, the results indicated that TzAgNPs exhibits considerable reduction in the generation of biofilms and spreading of virulence factors. Taken together, all the results indicated that TzAgNPs could be deemed to be a promising agent for the retardation of microbial biofilm that might assist to fight against infections linked to biofilm.
Keywords: Pseudomonas aeruginosa, TzAgNPs, antimicrobial, antibiofilm, virulence factor.
Microorganisms live in nature in two forms; one is planktonic where the microorganism lives distantly free from each other while the other is known as biofilm where the microorganism lives in clustered colony form (Garrett et al. 2008). Biofilm can develop on both biotic and abiotic substances and are found in almost every environment (Cortes et. al, 2011). Biofilm are the accumulation of microbial cells that are irreversibly adhered on a surface and confined within a layer of self-secreted extracellular polymeric substances (EPS) containing proteins, polysaccharides and DNA etc. (Sharma et al. 2015). The EPS forms an interconnecting three-dimensional polymer network of cohesive nature which helps the microorganism to adhere to the surface as well as imbomilize biofilms cells transiently. (Gupta et al. 2016). Microorganism adapts to the harsh condition by forming biofilm. It was reported that microbial biofilm is often involved in microbial pathogenesis (Vasudevan 2014). Biofilm promotes gene transfer in the horizontal direction among bacterial population that steers an expansion in the population of virulent strains (Lewis 2001). According to an estimation by the centre for disease control (CDC) and National Institutes of Health (Joo and Otto, 2012), bacterial biofilm are involved in a multitude of serious, chronic infections, 65 and 80 percent
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