Biosurfactant synergized with marine bacterial DNase disrupts polymicrobial biofilms.

Globally, the occurrence of biofilm associated infection has become an alarming menace to the medical fraternity because the thick exopolysaccharide layer encasing the biofilms makes the biofilm producing pathogens inherently resistant to antibiotics. Candida albicans, the most common pathogen among Candida spp. is the causative agent for superficial and invasive candidiasis. The morphological phase switching from yeast to hyphal form is one of the virulent traits of C. albicans critical for its pathogenicity. Owing to the emergence of antifungal resistance among this opportunistic fungus, there is a dire need for improvised alternative antifungal agents. In the present study, we have evaluated a biosurfactant from a marine bacterium for its biofilm disruption ability against C. albicans. This biosurfactant had the potential to disrupt biofilms as well as to inhibit the morphological transition from yeast to hyphae. In addition, this biosurfactant showed enhance disruption of mixed species biofilms of C. albicans and Staphylococcus epidermidis when combined with DNase isolated from marine bacteria. From the results obtained, it is evident that the biosurfactant could act as a potential antibiofilm agent against drug resistant C. albicans strains.

Chitosan-Coated Surgical Sutures Prevent Adherence and Biofilms of Mixed Microbial Communities.

Sutures are widely used materials for closing the surgical wounds, and being an inert material, sutures are often colonized with drug-resistant polymicrobial biofilms. Surgical site infection (SSI) is a hospital-acquired infection caused by bacteria and fungi specifically in the sutured sites. Although most of the currently available sutures possess antibacterial property, their ability to prevent biofilm colonization by polymicrobial communities is underexplored. So, the present study shows that extracted chitosan (EC) from crab shells prevented the adherence of Staphylococcus epidermidis and Candida albicans, the predominant members that exist as mixed species at the site of SSI. In comparison with a commercial chitosan, EC showed profound inhibition of slime formation and mixed species biofilm inhibition. Intriguingly, EC-coated sutures could inhibit the growth of both bacterial and fungal pathogens when comparing with a commercial triclosan-coated suture which was active only against the bacterial pathogen. Scanning electron microscopy results revealed inhibition of C. albicans hyphal formation by the EC-coated sutures that is a crucial virulence factor responsible for tissue invasiveness. Collectively, the results of the present study showed that EC from crab shells (discarded material as a recalcitrant biowaste) could be used as an alternative to combat drug-resistant biofilms which are the prime cause for SSIs.

Gallic Acid an Agricultural Byproduct Modulates the Biofilm Matrix Exopolysaccharides of the Phytopathogen Ralstonia solanacearum.

Ralstonia solanacearum is a soil-borne plant pathogen which causes wilt disease in economically important crops of the Solanaceae family in tropical and temperate regions. As biofilm formation is the major virulence factor in R. solanacearum, research inputs are necessary to identify natural biofilm inhibitors to mitigate virulence of this bacterium. Hence in the present work, the anti-biofilm potential of phytochemical compound gallic acid (GA) isolated from an agricultural byproduct (cashewnut shell) was investigated. Initially the Minimum inhibitory concentration (MIC) of crude extracts of cashewnut shell and coconut shell against R. solanacearum were investigated. The MIC of both the extracts were 400 µg/ml and their sub-MIC (200 µg/ml) inhibited biofilms in the range of 62-70% and 49-57%, respectively. As the cashewnut shell extract have higher biofilm inhibitory effect compared to coconut shell extract, we proceeded our further study by isolating the major compound GA from cashewnut shell by acid hydrolysate method. The sub-MIC of crude cashewnut shell extract inhibited 85% of young biofilms. The MIC of GA were observed at 3 mg/ml and sub-MIC (1.5 mg/ml) was found to eradicate 85% of mature biofilms which was confirmed by standard crystal violet assay and the biofilm reduction was further visualized under light microscopy and scanning electron microscopic images. Toxicity of GA was evaluated against R. solanacearum through XTT cell viability assay and found no antibacterial effect at sub-MIC. Additionally, it is confirmed with growth curve and time kill assays. Swimming and twitching motility were considered as an important virulence factors to invade plants and to block the xylem vessels. Therefore, sub-MIC of GA was found to inhibit both swimming and twitching motility of about 93% and 63% respectively. Anti-biofilm efficacy of GA was also worked well with tomato plant model where remarkable biofilm inhibition was found on treatment with GA before and after 24 h of infection with R. solanacearum. Hence GA will be an alternative, cheap source which is eco-friendly as well as novel source for the treatment of R. solanacearum biofilms and to prevent wilt disease in important crops.

Agro food by-products and essential oil constituents curtail virulence and biofilm of Vibrio harveyi.

Vibrio harveyi causes severe loss to the aquaculture industry due to its virulence, which is mediated by Quorum sensing (QS) and biofilm formation. In the current study, we have explored the anti-virulent properties and biofilm disruption ability of luteolin (extracted from coconut shell) and linalool against this important aquaculture pathogen. HPLC analysis of the methanolic extract of coconut shells revealed a single major peak which matched to the standard luteolin which was further elucidated by NMR studies. Further, luteolin and linalool were screened for their ability to inhibit biofilms and various quorum sensing mediated virulence factors of V. harveyi. The Minimum Inhibitory Concentration (MIC) of the two compounds was determined and the sub-inhibitory concentrations of the compounds were able to inhibit biofilm formation. Both the compounds disrupted about 60-70% mature biofilms, which was also visually observed by light microscopy. Both linalool and luteolin exhibited a significant reduction in the production of EPS and alginate in the biofilms matrix of V. harveyi which was confirmed by Scanning Electron Microscopy (SEM). Both compounds inhibited the swarming and swimming motility, the crucial quorum sensing (QS) mediated virulence of V. harveyi. The present study shows the presence of valuable polyphenolic compound like luteolin in coconut shells that are discarded as a waste. From the present study we envisage that luteolin and linalool can serve as potent anti-virulent agents to combat QS mediated infections against aquaculture pathogens.