Endophytic fungi from Passiflora incarnata: an antioxidant compound source.

Endophytes are considered one of the most important microbial resources for obtaining biomolecules of therapeutic use. Passiflora incarnata, widely employed by the pharmaceutical industry, shows therapeutic effects on anxiety, nervousness, constipation, dyspepsia and insomnia based on their antioxidant compounds. In this study, from 315 endophytic fungi isolated from P. incarnata leaves, 60 were selected to determinate presence of chemical constituents related with antioxidant activity, based on their production of soluble pigments. The promising fungi were evaluated specifically on their potential to produce phenolic compounds, flavonoids and for antioxidant activity. Five isolates significantly produced flavonoids and phenolic compounds in the ethyl acetate and n-Butanol extracts, also saponins and high antioxidant activity against the DPPH (2.2-diphenyl-1-picrylhydrazyl) free radical. A strain of Aspergillus nidulans var. dentatus (former Emericella dentata) was able to produce tannins as well; its butanolic extract was very similar than the BHT (butylated hydroxytoluene) (94.3% × 94.32%) and Rutin (95.8%) reference substances in the DPPH radical scavenging. Similarly, a Chaetomium strain exhibited 93.6% and 94.7% of antioxidant activity in their ethyl acetate and butanolic fractions, respectively. The chromatographic analysis of the ethyl acetate fraction from the Aspergillus strain revealed the production of orcinol (3.19%). Four-methoxymethylphenol (4.79%), sorbicillin (33.59%) and ergosterol (23.08%) was produced by Trichoderma longibrachiatum and isopropenyl-1,4-dimethyl-1,2,3,3a,4,5,6,7-octahydroazulene were found in two Fusarium oxysporum strains. The phytochemical screening showed that all analyzed fungi were able to produce a kind of secondary metabolite (phenols, flavonoids, tannins and/or saponins). The study shows a great underexplored potential for industrial application of P. incarnata endophytes.

Detoxification of sugarcane bagasse hydrolysate with different adsorbents to improve the fermentative process.

Second generation ethanol has the prospect of becoming an important bioenergy alternative. The development of this technology is associated with the lignocellulosic materials' use, with emphasis on agricultural and agroindustrial by-products from which fermentable sugar can be produced. The acid hydrolysis depolymerizes the hemicellulose releasing mainly xylose. Subsequently, the cellulose can be converted into glucose by enzymatic hydrolysis. However, the acid hydrolysis produces toxic compounds, such as furan derivatives, phenolics, and organic acids, which are harmful to fermentative microorganisms. This study investigated different acid concentrations in the sulfuric acid hydrolysis of sugarcane bagasse (1- 5% m/v) and the use of adsorbents with the prerogative to improve the acid hydrolysate (AH) quality for microbial ethanolic fermentation. Cell growth and fermentative yield of Saccharomyces cerevisiae (PE-2) and Scheffersomyces stipitis (NRRL Y-7124) were evaluated. AH was used as a source of pentoses (17.7 g L-1) and molasses (ME) sugarcane as source of hexoses (47 g L-1). The following adsorbents were used: activated charcoal, clay, hydrotalcite and active and inactive cells of PE-2 and NRRL Y-7124, at concentrations ranging (1 - 8% m/v). Results of cell growth and chemical characterization allowed to select the most effective adsorbents with emphasis for active cells that removed 66% furfural and 51% 5-(hydroxymethyl) furfural) (5-HMF) and alcoholic productivity of 23.5 g L-1 in AH and ME substrates, in the presence of mixed culture. These results indicate the application of active yeast cells in the detoxification of acid hydrolysates of the sugarcane bagasse previously to the fermentation.

Monitoring Heavy Metals in Royal Sun Medicinal Mushroom, Agaricus brasiliensis (Agaricomycetes).

Agaricus brasiliensis is popular because of its nutritional and medicinal properties. Brazil supplies this edible mushroom to international markets, where quality standards are very restrictive regarding the allowable concentrations of heavy metals, especially cadmium. In order to evaluate the accumulation of cadmium, chromium, and lead in A. brasiliensis, the fungus was grown as producers cultivate it, using plastic bags containing inoculated compost with soil as a covering. We analyzed the soil and compost using atomic absorption spectrophotometry to determine their physical and chemical properties and natural concentrations of the heavy metals cadmium, chromium, and lead. In addition to the metals naturally found in the soil and compost, 6.6 mgkg-1 cadmium chloride, 13.2 mgkg-1 chromium sulfate, and 1.2 mgkg-1 lead nitrate were added to the soil covering to simulate contaminated soils and to allow us to evaluate potential adsorption by the fungus. An analysis of the fruiting bodies after fructification showed accumulation of the heavy metals, with the greatest amount in those fruiting bodies that had been contaminated with cadmium. Furthermore, the contaminated fruiting bodies had the least raw protein content, were physically deformed, and exhibited variations in the results of Fourier transform infrared spectroscopy analyses.

Assessment of the cytotoxic, genotoxic and mutagenic effects of the commercial black dye in Allium cepa cells before and after bacterial biodegradation treatment.

The present study evaluated the cytotoxic, genotoxic and mutagenic actions of different concentrations (50 and 200 µg/L) of BDCP (Black Dye Commercial Product) used by textile industries, before and after bacterial biodegradation, by the conventional staining cytogenetic technique and NOR-banding in Allium cepa cells. Differences in the chromosomal and nuclear aberrations and alterations in the number of nucleoli were observed in cells exposed to BDCP with and without the microbial treatment. The significant frequencies of chromosome and nuclear aberrations noted in the tests with bacterially biodegraded BDCP indicate that the metabolites generated by degradation are more genotoxic than the chemical itself. Losses of genetic material characterize a type of alteration that was mainly associated with the action of the original BDCP, whereas chromosome stickiness, nuclear buds and binucleated cells were the aberrations that were preferentially induced by BDCP metabolites after biodegradation. The significant frequencies of cell death observed in the tests with biodegraded BDCP also show the cytotoxic effects of the BDCP metabolites. The reduction in the total frequency of altered cells after the recovery treatments showed that the test organism A. cepa has the ability to recover from damage induced by BDCP and its metabolites after the exposure conditions are normalized.

Water Quality Index (WQI) of Jaguari and Atibaia Rivers in the region of Paulínia, São Paulo, Brazil.

Due to the concern with the quality of hydric resources, the monitoring is essential to evaluate and identify the anthropogenic and environmental interferences in a quantitative and qualitative level. In order to assist the interpretation of the water status of hydric bodies, the gathering of analytical data often considers the Water Quality Index (WQI). This index transforms technical information in description of the water quality status, highlighting the effectiveness of its use and guiding the decision-making process when necessary. The aim of this research is to assess the water quality of a region in São Paulo State, Brazil, by means of the WQI of Jaguari and Atibaia Rivers. The period of intense drought, which affected the Brazilian southeast in 2014, was evaluated and compared to the mean values recorded from October 2009 to March 2015, correlating the values of Escherichia coli and the biochemical oxygen demand (BOD). One hundred nine samples were collected, being 53 from dry seasons, between October and March, and 56 from rainy seasons, between April and September. The WQIs in Jaguari and Atibaia Rivers, during dry season, were of 42.2 (medium) and 36.7 (bad), respectively. The same pattern was registered for the dry season, for both rivers, with indices values of 40.1 for Jaguari River (medium) and 34.9 for Atibaia River (bad). This research presented the need of an effective evaluation of the environmental quality and preservation by competent organs for both rivers. Due to the detected conditions, special attention should be given to the Atibaia River.

Application of micronucleus test and comet assay to evaluate BTEX biodegradation.

The BTEX (benzene, toluene, ethylbenzene and xylene) mixture is an environmental pollutant that has a high potential to contaminate water resources, especially groundwater. The bioremediation process by microorganisms has often been used as a tool for removing BTEX from contaminated sites. The application of biological assays is useful in evaluating the efficiency of bioremediation processes, besides identifying the toxicity of the original contaminants. It also allows identifying the effects of possible metabolites formed during the biodegradation process on test organisms. In this study, we evaluated the genotoxic and mutagenic potential of five different BTEX concentrations in rat hepatoma tissue culture (HTC) cells, using comet and micronucleus assays, before and after biodegradation. A mutagenic effect was observed for the highest concentration tested and for its respective non-biodegraded concentration. Genotoxicity was significant for all non-biodegraded concentrations and not significant for the biodegraded ones. According to our results, we can state that BTEX is mutagenic at concentrations close to its water solubility, and genotoxic even at lower concentrations, differing from some described results reported for the mixture components, when tested individually. Our results suggest a synergistic effect for the mixture and that the biodegradation process is a safe and efficient methodology to be applied at BTEX-contaminated sites.

In vitro susceptibility of environmental isolates of Exophiala dermatitidis to five antifungal drugs.

Several dematiaceous fungi frequently isolated from nature are involved in cases of superficial lesions to lethal cerebral infections. Antifungal susceptibility data on environmental and clinical isolates are still sparse despite the advances in testing methods. The objective of this study was to examine the activities of 5-flucytosine, amphotericin B, itraconazole, voriconazole and terbinafine against environmental isolates of Exophiala strains by minimum inhibition concentration (MIC) determination. The strains were obtained from hydrocarbon-contaminated soil, ant cuticle and fungal pellets from the infrabuccal pocket of attine gynes. Broth microdilution assay using M38-A2 reference methodology for the five antifungal drugs and DNA sequencing for fungal identification were applied. Terbinafine was the most active drug against the tested strains. It was observed that amphotericin B was less effective, notably against Exophiala spinifera, also studied. High MICs of 5-flucytosine against Exophiala dermatitidis occurred. This finding highlights the relevance of studies on the antifungal resistance of these potential opportunistic species. Our results also contribute to a future improvement of the standard methods to access the drug efficacy currently applied to black fungi.

BTEX biodegradation by bacteria from effluents of petroleum refinery.

Groundwater contamination with benzene, toluene, ethylbenzene and xylene (BTEX) has been increasing, thus requiring an urgent development of methodologies that are able to remove or minimize the damages these compounds can cause to the environment. The biodegradation process using microorganisms has been regarded as an efficient technology to treat places contaminated with hydrocarbons, since they are able to biotransform and/or biodegrade target pollutants. To prove the efficiency of this process, besides chemical analysis, the use of biological assessments has been indicated. This work identified and selected BTEX-biodegrading microorganisms present in effluents from petroleum refinery, and evaluated the efficiency of microorganism biodegradation process for reducing genotoxic and mutagenic BTEX damage through two test-systems: Allium cepa and hepatoma tissue culture (HTC) cells. Five different non-biodegraded BTEX concentrations were evaluated in relation to biodegraded concentrations. The biodegradation process was performed in a BOD Trak Apparatus (HACH) for 20 days, using microorganisms pre-selected through enrichment. Although the biodegradation usually occurs by a consortium of different microorganisms, the consortium in this study was composed exclusively of five bacteria species and the bacteria Pseudomonas putida was held responsible for the BTEX biodegradation. The chemical analyses showed that BTEX was reduced in the biodegraded concentrations. The results obtained with genotoxicity assays, carried out with both A. cepa and HTC cells, showed that the biodegradation process was able to decrease the genotoxic damages of BTEX. By mutagenic tests, we observed a decrease in damage only to the A. cepa organism. Although no decrease in mutagenicity was observed for HTC cells, no increase of this effect after the biodegradation process was observed either. The application of pre-selected bacteria in biodegradation processes can represent a reliable and effective tool in the treatment of water contaminated with BTEX mixture. Therefore, the raw petroleum refinery effluent might be a source of hydrocarbon-biodegrading microorganisms.

Dynamics and control strategies for a butanol fermentation process.

In this work, mathematical modeling was employed to assess the dynamic behavior of the flash fermentation process for the production of butanol. This process consists of three interconnected units as follows: fermentor, cell retention system (tangential microfiltration), and vacuum flash vessel (responsible for the continuous recovery of butanol from the broth). Based on the study of the dynamics of the process, suitable feedback control strategies [single input/single output (SISO) and multiple input/multiple output (MIMO)] were elaborated to deal with disturbances related to the process. The regulatory control consisted of keeping sugar and/or butanol concentrations in the fermentor constant in the face of disturbances in the feed substrate concentration. Another objective was the maintenance of the proper operation of the flash tank (maintenance of the thermodynamic equilibrium of the liquid and vapor phases) considering that oscillations in the temperature in the tank are expected. The servo control consisted of changes in concentration set points. The performance of an advanced controller, the dynamic matrix control, and the classical proportional-integral controller was evaluated. Both controllers were able to regulate the operating conditions in order to accommodate the perturbations with the lowest possible alterations in the process outputs. However, the performance of the PI controller was superior because it showed quicker responses without oscillations.

Action mechanisms of petroleum hydrocarbons present in waters impacted by an oil spill on the genetic material of Allium cepa root cells.

Chromosomal aberration (CA) assays have been widely used, not only to assess the genotoxic effects of chemical agents, but also to evaluate their action mechanisms on the genetic material of exposed organisms. This is of particular interest, since such analyses provide a better knowledge related to the action of these agents on DNA. Among test organisms, Allium cepa is an outstanding species due to its sensitivity and suitable chromosomal features, which are essential for studies on chromosomal damage or disturbances in cell cycle. The goal of the present study was to analyze the action mechanisms of chemical agents present in petroleum polluted waters. Therefore, CA assay was carried out in A. cepa meristematic cells exposed to the Guaecá river waters, located in the city of São Sebastião, SP, Brazil, which had its waters impacted by an oil pipeline leak. Analyses of the aberration types showed clastogenic and aneugenic effects for the roots exposed to the polluted waters from Guaecá river, besides the induction of cell death. Probably all the observed effects were induced by the petroleum hydrocarbons derived from the oil leakage.