Physiological and Ecological Aspects of Chlorella sorokiniana (Trebouxiophyceae) Under Photoautotrophic and Mixotrophic Conditions.

Mixotrophy is a metabolic strategy in which an organism is autotrophic and heterotrophic simultaneously. Considering that the aquatic environment provides several organic sources of carbon, it is probably common for microalgae to perform mixotrophy and not only photoautotrophy, but little is known about microalgae mixotrophy. The present work aimed at investigating the growth, photosynthetic activity, morphology, and biochemical composition of the microalga Chlorella sorokiniana in mixotrophic and photo-mixotrophic conditions, comparing it with photoautotrophy. The results showed pH changes after glucose addition, reaching pH 11.62 in mixotrophic and 10.47 in sequential photo-mixotrophic cultures, which limited the microalgal growth. Highest biomass was obtained in the mixotrophic culture in comparison with the sequential photo-mixotrophic one. Rapid light saturation curves showed that α (photosynthetic efficiency, 1.69) and relative electron transport rate (rETR; 565.61) were higher in the mixotrophic cultures, whereas the highest Ik (irradiance saturation, 386.68) was obtained in the photoautotrophic ones. In the sequential photo-mixotrophic cultures, photosynthetic activity varied during glucose consumption, decreasing the maximum quantum yield Fv/Fm after glucose addition, indicating change in metabolism, from photoautotrophy to mixotrophy by the microalga. The results showed that the mixotrophic cultures had higher production of chlorophyll a (6.26 mg mL-1), cell density (6.62 × 107 cell mL-1), and lipids (0.06 pg µm-3). Sequential photo-mixotrophic cultures showed the highest biovolume (360.5 µm3 cell-1) and total carbohydrates (0.026 pg µm-3). The protein concentration was 3.2 and 2.4 times higher in photoautotrophy and photo-mixotrophic growth, respectively, than in mixotrophy, but lipids were three times higher under mixotrophy. The biochemical changes we observed indicate that the microalga's plasticity in face of new environmental characteristics, such as the presence of organic carbon, can change the flow of energy through natural ecosystems.

Different Photoresponses of Microorganisms: From Bioinhibition to Biostimulation.

The effective treatment of antimicrobial modalities continues to be a serious challenge, mainly due to the increasing number of multidrug resistance pathogenic microorganisms. Microbial bioinhibition is an alternative method that has shown to be effective. This study investigated and described the effect of the visible light on five different microorganisms. The studied groups were composed by the species Acanthamoeba polyphaga, Candida albicans, Mycobacterium massiliense, Pseudomonas aeruginosa, and Staphylococcus aureus. These microorganisms were analyzed after six light doses exposition with three different wavelengths: 450, 520, and 630 nm. The present study indicates two different behaviors: bioinhibition and/or biostimulation. The bioinhibition effect was calculated using different percentages of the microorganism population, compared to the control group, in which the maximum value corresponds to 94% growth inhibition. The biostimulation effect was evaluated by the microorganism population increment for specific light doses. Our results showed a 132% population growth as the maximum value. These results were assessed by variance analysis. The Tukey's test was used for differentiating or comparing, depending on the circumstances. The obtained results suggested a visible light phototherapeutic effect that could be used as a microorganism inactivation method for the studied microorganisms. In some approaches, the biostimulation effect might also be a very interesting effect to be considered. This study supports the relevance of understanding the important role that phototherapy plays as a useful method for microbiological control studies and applications.

Photoinactivation of microorganisms using bacteriochlorins as photosensitizers.

In recent years, some microorganisms have shown resistance to conventional treatments. Considering this increase in resistant pathogens, treatment alternatives are needed to promote greater treatment efficiency. In this sense, antimicrobial photodynamic therapy (aPDT) has been an alternative treatment. This technique uses a photosensitizer that is activated by light with a specific wavelength producing reactive species, leading to the death of pathogenic microorganisms. In this study, bacteriochlorophyll derivatives such as bacteriochlorin metoxi (Bchl-M) and bacteriochlorin trizma (Bchl-T) obtained from purple bacterium (Rhodopseudomonas faecalis), were evaluated as photosensitizers in the aPDT. Photodynamic inactivation (PDI) of the microorganisms Staphylococcus aureus, Micrococcus luteus, Candida albicans and Pseudomonas aeruginosa was investigated with both bacteriochlorins (Bchl-M and Bchl-T) at different concentrations (1, 15 and 30 µM for S. aureus; 1, 15, 30, 45, 60 and 75 µM for M. luteus; 30, 60, 90, 105, 120 and 150 µM for C. albicans; and 200 µM for P. aeruginosa) and different doses of light (20 and 30 J/cm2 for S. aureus and M. luteus; 30 and 45 J/cm2 for C. albicans; and 45 J/cm2 for P. aeruginosa) to inactivate them. Both photosensitizers showed good activation against S. aureus and for M. luteus, we observed the inactivation of these microorganisms at approximately 3 log, showing to be a good photosensitizers for these microorganisms.

Evaluation of filter media covered with spun fibres and containing thyme essential oil with antimicrobial properties.

Inhalation of bioaerosols has been linked to many health problems. Filter media impregnated with antimicrobial material can provide effective removal and inactivation of bioaerosols. In this study, fibres were spun on a substrate by centrifugal spinning, obtaining filter media denoted 5THY and THY. Thyme essential oil was used as an antimicrobial agent. For 5THY, the thyme essential oil was added to the polyacrylonitrile (PAN) solution, while for THY, it was sprayed onto the medium after the fibres had been produced. The THY medium presented a higher collection efficiency, compared to the substrate or 5THY, with efficiencies of 99% for microparticles and 58% for nanoparticles. Using the plaque assay method, THY provided the highest reductions of the bacteria Escherichia coli and Staphylococcus aureus, with efficiency of 99.999%. The findings demonstrated that filter media covered with spun fibres and containing thyme essential oil provided excellent antimicrobial action and filtration performance.

Bactericidal activity and biofilm inhibition of F18 bioactive glass against Staphylococcus aureus.

Antimicrobial treatment failure has been increasing at alarming rates. In this context, the bactericidal properties of biocompatible antimicrobial agents have been widely studied. F18 is a recently developed bioactive glass that presents a much wider working range when compared to other bioactive glasses, a feature that allows it to be used for coating metallic implants, sintering scaffolds or manufacturing fibers for wound healing applications. The aim of this study was to investigate the in vitro bactericidal and anti-biofilm activity of F18 glass as a powder and as a coating on steel samples, and to explore the effects of its dissolution products at concentrations from 3 mg/mL to 50 mg/mL against the Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA) biofilms. Furthermore, we intend to verify whether changes in the medium pH could influence the bactericidal activity of F18. The results indicated that F18 presented bactericidal activity in preformed S. aureus and MRSA biofilms, reducing more than 6 logs of the viable cells that remained in contact with 50 mg/mL for 24 h. Moreover, an anti-biofilm activity was observed after 12 h of direct contact, with a drop of more than 6 logs of the viable bacterial population. Neutralization of the F18 solution pH decreased its bactericidal efficacy. These results indicate that the F18 glass could be considered as an alternative material for controlling and treating infections by S. aureus.

Photodynamic therapy with a new bacteriochlorin derivative: Characterization and in vitro studies.

Photodynamic therapy presents a therapeutic choice that can be utilized to treat diverse neoplasms. In this technique, the critical element is a photosensitive molecule that absorbs light energy and transfers it to molecular oxygen or biological molecules to form reactive oxygen species, thus inducing irreversible damage to target cells and ultimately leading to cell death. Bacteriochlorin derivatives are employed as photosensitizers (PSs), possessing light-absorbing capacity in the near-infrared region. The objective of this study was to prepare a semi-synthetic bacteriochlorin from Rhodopseudomonas faecalis and adding Trizma® to improve solubility. Cell viability tests, flow cytometry (apoptotic and necrotic cells were identified by Annexin V and propidium iodide), and confocal microscopy were used to evaluate the photoactivity of bacteriochlorin-Trizma (Bchl-T) in fibroblast (HFF-1-control cells) and breast cancer (MCF-7 cells-target cells) cells. At concentrations above 0.5 µM, Bchl-T demonstrated 80 % cell death, presenting the highest PS interaction (via fluorescence microscopy) with lysosomes, mitochondria, and the endoplasmic reticulum; the cell death type was revealed as apoptosis (via cytometry). Our findings indicated the suitability of Bchl-T for future application in photodynamic therapy against cancer cells by inducing apoptosis.

Oral Decontamination of Orthodontic Patients Using Photodynamic Therapy Mediated by Blue-Light Irradiation and Curcumin Associated with Sodium Dodecyl Sulfate.

OBJECTIVE: The aim of this study was to investigate the effects of the antimicrobial photodynamic therapy (aPDT) using the association of curcumin with the surfactant sodium dodecyl sulfate (SDS) for oral decontamination in orthodontic patients. BACKGROUND DATA: The installation of the orthodontic appliances promotes an increase in the retentive area that is available for microbial aggregation and makes difficult the oral health promotion. However, aPDT is one possible approach that is used for the reduction of oral microbial load. MATERIALS AND METHODS: Twenty-four patients (n = 24) were randomly distributed into four groups: Light group: which was treated only with the blue light, no drug; PDT group, which was treated with curcumin and blue light; PDT + S group, which was treated with curcumin plus surfactant and irradiated with blue light; and Chlorhex group, which was treated with chlorhexidine. The photosensitizer agent was prepared by adding 0.1% of SDS to a curcumin solution of 1 g/L. Two distinct LED devices emitting blue light (450 ± 10 nm) were used as follows: extra-oral irradiation (200 mW, 80 mW/cm(2), 36 J and 14 J/cm(2)) and intra-oral irradiation (1200 mW, 472 mW/cm(2), 216 J and 85 J/cm(2)).The collection of nonstimulated saliva (n = 3; 3 mL/collection) was performed at the following steps: (1) immediately before swishing (curcumin, chlorhexidine, or water); (2) after swishing; and (3) after performing aPDT treatments. The colony-forming units (CFU) were counted visually, and the values were adjusted to CFU/mL. RESULTS: There was significant Log reduction for PDT (from 6.33 ± 0.92 to 5.78 ± 0.96, p < 0.05), PDT + S (from 5.44 ± 0.94 to 3.83 ± 0.71, p < 0.01), and Chlorhex (from 5.89 ± 0.97 to 2.55 ± 1.80, p < 0.01) groups. The survival rate was significantly reduced in both PDT + S and Chlorhex groups compared with all situations (p < 0.05). However, there was no significant difference between PDT + S and Chlorhex groups (p ≥ 0.05). CONCLUSIONS: These results indicate that when associated with the surfactant SDS, the aPDT can be used as an adjutant and a convenient agent to promote the oral decontamination in clinical practice.

Addition of silver nanoparticles to composite resin: effect on physical and bactericidal properties in vitro.

The objectives of this study were to evaluate physical properties and antibacterial activity of a light-activated composite modified with silver nanoparticles. Discs were produced with unmodified resin (control group - CG) and modified resin with silver nanoparticles at two concentrations, 0.3% wt (MR03) and 0.6% wt (MR06). Streptococcus mutans and Lactobacillus acidophilus biofilms were induced in vitro by incubation of discs in a 20% sucrose medium, followed by sonication and counting of viable cells after 1, 4 and 7 days (n=9). The arithmetic roughness of all three groups was evaluated by atomic force microscopy (n=9). Compression assay was conducted in all groups to measure the compressive strength at failure and elasticity modulus (n=5). Data were subjected to ANOVA and Tukey's tests (α=0.05%). At all three time points the number of viable cells was statistically lower for MR03 and MR06 compared with CG, for both specimens. MR03 and MR06 showed no significant differences. Microscopic analysis demonstrated no significant differences for roughness among the three groups (p>0.05). The MR03 was stronger to compression than CG, and MR06 was statistically lower than CG and MR03. It was concluded that the MR03 were less conducive to biofilm growth, without compromising the strength in compression and surface roughness.