RESUMEN
Candida is one of the most prevalent fungal pathogens in clinical settings which form antibiotic-resistant biofilms on biomedical devices. Hence, there is a need for non-antimicrobial alternatives to combat these infections. The present study investigates the anti-biofilm effect of marine bacterial DNase by targeting the eDNA present in the biofilms of Candida spp. A strain of Vibrio alginolyticus (AMSII) which showed enhanced DNase activity was isolated from marine sediment. Treatment of young and mature Candida biofilms with purified marine bacterial DNase (MBD) caused a 60-80% reduction in biofilm biomass, similar to treatment with DNase I from Bovine pancreas. Scanning electron microscopy showed that MBD significantly reduced the formation of biofilms on urinary catheters and more importantly prevented the virulent yeast to hyphae dimorphic switch in C. albicans. The present study identified a potential non-antibiotic alternative therapy to eradicate Candida biofilms and can be used to develop enzyme fabricated antifouling indwelling medical devices.
Asunto(s)
Antifúngicos/farmacología , Biopelículas/efectos de los fármacos , Candida albicans/efectos de los fármacos , Desoxirribonucleasas/farmacología , Animales , Antifúngicos/aislamiento & purificación , Biopelículas/crecimiento & desarrollo , Candida/efectos de los fármacos , Candida/patogenicidad , Candida albicans/patogenicidad , Bovinos , Desoxirribonucleasas/aislamiento & purificación , Microscopía Electrónica de Rastreo , Catéteres Urinarios/microbiología , Vibrio alginolyticus/enzimología , VirulenciaRESUMEN
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.
Asunto(s)
Antibacterianos/farmacología , Biopelículas/efectos de los fármacos , Alimentos , Aceites Volátiles/farmacología , Extractos Vegetales/farmacología , Vibrio/efectos de los fármacos , Virulencia/efectos de los fármacos , Monoterpenos Acíclicos/aislamiento & purificación , Monoterpenos Acíclicos/farmacología , Alginatos/análisis , Acuicultura , Supervivencia Celular/efectos de los fármacos , Hidroxibenzoatos/farmacología , Luteolina/aislamiento & purificación , Luteolina/farmacología , Pruebas de Sensibilidad Microbiana , Percepción de Quorum/efectos de los fármacos , Vibrio/crecimiento & desarrollo , Vibriosis , Factores de VirulenciaRESUMEN
Multidrug-resistant (MDR) Staphylococcus aureus is a major cause of biofilm-associated and indwelling device related infections. The present study explores the anti-virulent and antibiofilm potency of chitosan extracted from the shells of the marine crab Portunus sanguinolentus against Methicillin Resistant Staphylococcus aureus (MRSA). The chemical characterization results revealed that the extracted chitosan (EC) has structural analogy to that of a commercial chitosan (CC). The extracted chitosan was found to be effective in reducing the staphyloxanthin pigment, a characteristic virulence feature of MRSA that promotes resistance to reactive oxygen species. Furthermore, Confocal laser scanning microscope (CLSM) revealed that EC exhibited a phenomenal dose dependent antibiofilm efficacy against mature biofilms of the standard as well as clinical MRSA strains and Scanning Electron Microscopy (SEM) confirmed EC had a higher efficacy in disrupting the thick Exopolysaccharide (EPS) layer than CC. Additionally, EC and CC did not have any cytotoxic effects when tested on lung epithelial cell lines. Thus, the study exemplifies the anti-virulent properties of a marine bioresource which is till date discarded as a biowaste.
Asunto(s)
Antibacterianos/farmacología , Biopelículas/efectos de los fármacos , Quitosano/aislamiento & purificación , Quitosano/farmacología , Crustáceos/química , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Xantófilas/biosíntesis , Acetilación , Animales , Aniones , Adhesión Bacteriana/efectos de los fármacos , Cationes , Espectroscopía de Resonancia Magnética , Staphylococcus aureus Resistente a Meticilina/crecimiento & desarrollo , Staphylococcus aureus Resistente a Meticilina/metabolismo , Pruebas de Sensibilidad Microbiana , Microscopía Electrónica de Rastreo , Espectroscopía Infrarroja por Transformada de Fourier , Difracción de Rayos XRESUMEN
Pseudomonas aeruginosa is a nosocomial pathogen colonizing patients with chronic infectious diseases and has gained resistance to all the known broad spectrum antibiotics available today. The present study showcases the antibiofilm potential of an essential oil (EO) from an underexplored Cinnamomum species namely, C. tamala, against P. aeruginosa biofilms. Furthermore, the synergistic effects of the EO along with a commercially available DNase (DNaseI) and a DNase (MBD) isolated from a marine bacterium were explored for its antibiofilm activity. The results showed that the synergized action has maximum efficacy in inhibiting young and preformed biofilms. The synergized effect of EO and DNaseI showed 70% inhibition against matured biofilms of P. aeruginosa. The EO from C. tamala also showed quorum sensing inhibitory potential as it could inhibit the swarming motility behavior of P. aeruginosa. The synergistic action of EO and DNases offers a novel alternate therapeutic strategy for combating P. aeruginosa biofilm associated infections.