RESUMO
Light biotechnology is a promising tool for enhancing recalcitrant compounds biodegradation. Xenobiotics can cause a significant impact on the quality of the results achieved by sewage treatment systems due to their recalcitrance and toxicity. The optimization of bioremediation and industrial processes, aiming to increase efficiency and income is of great value. The aim of this study was to accelerate and optimize the hydrolysis of Remazol Brilliant Blue R by photo stimulating a thermophilic bacterial consortium. Three experimental groups were studied: control group; LED Group and Laser Group. The control group was exposed to the same conditions as the irradiated groups, except exposure to light. The samples were irradiated in Petri dishes with either a Laser device (λ660 nm, CW, θâ¯=â¯0.04â¯cm2, 40â¯mW, 325â¯s, 13â¯J/cm2) or by a LED prototype (λ632⯱â¯2â¯nm, CW, θâ¯=â¯0.5â¯cm2, 145â¯mW, 44â¯s, 13â¯J/cm2). We found that, within 48-h, statistically significant differences were observed between the irradiated and the control groups in the production of RNA, proteins, as well as in the degradation of the RBBR. It is concluded that, both Laser and LED light irradiation caused increased cellular proliferation, protein production and metabolic activity, anticipating and increasing the catabolism of the RBBR. Being the economic viability a predominant aspect for industrial propose our results indicates that photo stimulation is a low-cost booster of bioprocesses.
Assuntos
Antraquinonas/química , Processos Fotoquímicos , Xenobióticos/metabolismo , Antraquinonas/metabolismo , Antraquinonas/efeitos da radiação , Biodegradação Ambiental , Custos e Análise de Custo , Hidrólise , Lasers , Luz , Consórcios Microbianos/efeitos da radiação , Xenobióticos/efeitos da radiaçãoRESUMO
BACKGROUND: Candida tropicalis is increasingly becoming among the most commonly isolated pathogens causing fungal infections with an important biofilm-forming capacity. PURPOSE: This study addresses the antifungal effect of rubiadin (AQ1) and rubiadin 1-methyl ether (AQ2), two photosensitizing anthraquinones (AQs) isolated from Heterophyllaea pustulata, against C. tropicalis biofilms, by studying the cellular stress and antioxidant response in two experimental conditions: darkness and irradiation. The combination with Amphotericin B (AmB) was assayed to evaluate the synergic effect. STUDY DESIGN/METHODS: Biofilms of clinical isolates and reference strain of Candida tropicalis were treated with AQs (AQ1 or AQ2) and/or AmB, and the biofilms depletion was studied by crystal violet and confocal scanning laser microscopy (CSLM). The oxidant metabolites production and the response of antioxidant defense system were also evaluated under dark and irradiation conditions, being the light a trigger for photo-activation of the AQs. The Reactive Oxygen Species (ROS) were detected by the reduction of Nitro Blue Tetrazolium test, and Reactive Nitrogen Intermediates (RNI) by the Griess assay. ROS accumulation was also detected inside biofilms by using 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) probe, which was visualized by CSLM. Superoxide dismutase (SOD) activity and the total antioxidant capacity of biofilms were measured by spectrophotometric methods. The minimun inhibitory concentration for sessile cells (SMIC) was determined for each AQs and AmB. The fractional inhibitory concentration index (FICI) was calculated for the combinations of each AQ with AmB by the checkerboard microdilution method. RESULTS: Biofilm reduction of both strains was more effective with AQ1 than with AQ2. The antifungal effect was mediated by an oxidative and nitrosative stress under irradiation, with a significant accumulation of endogenous ROS detected by CSLM and an increase in the SOD activity. Thus, the prooxidant-antioxidant balance was altered especially by AQ1. The best synergic combination with AmB was also obtained with AQ1 (80.5%) (FICI=0.74). CONCLUSION: Under irradiation, the oxidative stress was the predominant effect, altering the prooxidant-antioxidant balance, which may be the cause of the irreversible cell injury in the biofilm. Our results showed synergism of these natural AQs with AmB. Therefore, the photosensitizing AQ1 could be an alternative for the Candida infections treatment, which deserves further investigation.