Impact of lead (Pb2+) on the growth and biological activity of Serratia marcescens selected for wastewater treatment and identification of its zntR gene-a metal efflux regulator.

Microorganisms isolated from contaminated areas play an important role in bioremediation processes. They promote heavy metal removal from the environment by adsorbing ions onto the cell wall surface, accumulating them inside the cells, or reducing, complexing, or precipitating these substances in the environment. Microorganism-based bioremediation processes can be highly efficient, low-cost and have low environmental impact. Thus, the present study aimed to select Pb2+-resistant bacteria and evaluate the growth rate, biological activity, and the presence of genes associated with metal resistance. Serratia marcescens CCMA 1010, that was previously isolated from coffee processing wastewater, was selected since was able to growth in Pb2+ concentrations of up to 4.0 mM. The growth rate and generation time did not differ from those of the control (without Pb2+), although biological activity decreased in the first hour of exposure to these ions and stabilized after this period. The presence of the zntR, zntA and pbrA genes was analysed, and only zntR was detected. The zntR gene encodes a protein responsible for regulating the production of ZntA, a transmembrane protein that facilitates Pb2+ extrusion out of the cell. S. marcescens CCMA 1010 demonstrated a potential for use as bioindicator that has potential to be used in bioremediation processes due to its resistance to high concentrations of Pb2+, ability to grow until 24 h of exposure, and possession of a gene that indicates the existence of mechanisms associated with resistance to lead (Pb2+).

Volatile compounds for biotechnological applications produced during competitive interactions between yeasts and fungi.

Fungi, yeasts and bacteria produce volatile compounds during their metabolism. In this study, the volatile compounds produced by yeast strains (Saccharomyces cerevisiae and Rhodotorula mucilaginosa) and fungal strains (Aspergillus carbonarius and Aspergillus ochraceus) during competitive interactions were investigated by solid-phase microextraction coupled with gas chromatography-mass spectrometry. Fifty-six volatile compounds were identified representing alcohols, aldehydes, esters, ketones, aromatic compounds, acids, furans, phenols, and nitrogen compounds, being the largest amount in the class of esters and alcohols. Eight compounds were identified only in interactive culture conditions such as 2-amino-1-propanol, isopropylamine, dimethylamine, pentyl propanoate, ethyl-2-aminopropanoate, acetone, oxalic acid, and ß-elemene and five of these were produced in cocultures including A. carbonarius. These will be developed for future biotechnological applications such as in the pharmaceutical and biological industry to produce drugs. Antimicrobial and antifungal activities; Solvent and herbicide; flavoring ingredient; solvent, plastic synthesis, nail polish remover and thinner, pesticide and herbicide; important in the complexation of minerals in the soil; and plant-environment interactions, defending predators, pathogens, and competitors.

Bio-hydrolysis of used soybean oil: environmental-friendly technology using microbial consortium.

In this work, strains of Bacillus subtilis were inoculated in consortium with Rhodotorula mucilaginosa into spent soy oil as aiming to biological treatment and low-cost reuse. The microorganisms were previously isolated and selected for the lipolytic capacity of the alperujo residue generated during the processing of olive oil. For fermentation, bioassays containing Rhodotorula mucilaginosa isolated from alperujo and Candida rugosa CCMA 00371, both co-inoculated with Bacillus subtilis CCMA 0085 in medium containing (% w/v) 0.075 glucose and 0.375 (NH4)3 PO4 in 75 mL of water and 75 mL of spent soy oil. Despite the low biomass productivity, it has favorable characteristics to be used in animal feed supplementation. Spent soy oil was used as a carbon source proven by Bartha respirometer. The strains of R. mucilaginosa UFLA RAS 144 and B. subtilis CCMA 0085 are promising inoculants for oil degradation and can be applied in a waste treatment system.

Protocol to select efficient microorganisms to treat coffee wastewater.

The coffee processing wastewater (CPWW) requires treatment before being disposed of in the environment or reused due to its high organic and inorganic composition and a low pH. The indigenous microbiota from CPWW is highly diverse and could be selected as inoculums in treatment waste plants. Considering the physico-chemical characteristics of wastewater coffee, we elaborate on steps to select the microbial consortium that showed positive impact via decreasing the pollutant parameters of this effluent. The effectiveness was confirmed using wastewater from different origins with different chemical characteristics. A bacterial consortium composed by Serratia marcescens CCMA 1010 and CCMA 1012, Corynebacterium flavescens CCMA 1006, and Acetobacter indonesiensis CCMA 1002 was selected as the inoculums-based phenotypic assays. The mixed inoculum showed a highly active population (11.18 log CFU mL-1), promoting an 85% decrease in biochemical oxygen demand and a 60% decrease in chemical oxygen demand. There was also an 80% reduction in phosphorus and nitrogen. The final pH changed from 6.0 to 7.5. Additionally, the eco-toxicity using Daphnia similis was reduced by more than 59%. The microbial inoculum was efficient in the biological treatment in CPWWs, demonstrating the efficiency and robustness of the selected strains, independent of the physico-chemical characteristics of wastewater.

2-Phenylethanol and 2-phenylethylacetate production by nonconventional yeasts using tequila vinasses as a substrate.

Vinasses from the tequila industry are wastewaters with highly elevated organic loads. Therefore, to obtain value-added products by yeast fermentations, such as 2-phenylethanol (2-PE) and 2-phenylethylacetate (2-PEA), could be interesting for industrial applications from tequila vinasses. In this study, four yeasts species (Wickerhamomyces anomalus, Candida glabrata, Candida utilis, and Candida parapsilosis) were evaluated with two different chemically defined media and tequila vinasses. Differences in the aroma compounds production were observed depending on the medium and yeast species used. In tequila vinasses, the highest concentration (65 mg/L) of 2-PEA was reached by C. glabrata, the inhibitory compounds decreased biomass production and synthesis of 2-PEA, and biochemical and chemical oxygen demands were reduced by more than 50 %. Tequila vinasses were suitable for the production of 2-phenylethylacetate by the shikimate pathway. A metabolic network was developed to obtain a guideline to improve 2-PE and 2-PEA production using flux balance analysis (FBA).

Assessing the efficiency in assisted depuration of coffee processing wastewater from mixed wild microbial selected inoculum.

This work evaluated the efficiency of bacterial bio-augmentation to the biological treatment of coffee processing wastewater (CPWW) in a pilot wastewater treatment plant (WTP). Biochemical oxygen demand (BOD) and chemical oxygen demand (COD) values were the basis for the treatment efficiency. Serratia marcescens CCMA 1010 and CCMA 1013, Corynebacterium flavescens CCMA 1006 and Acetobacter indonesiensis CCMA 1002 were previously selected. The microbial cocktail was inoculated and persisted in CPWW during all treatments. The richness of wild species was a little altered over time and up to nine species were found in each sampled season. The microbiota composition presented variation of a total of 13 species, despite the inoculation of the microbial inoculum. The biodegradability index of effluent, close to 0.5, was favourable to biological treatment. The pollution parameters of CPWW were decreased in function of the variation of community composition and microbial activity. The greatest reduction of BOD (~ 33%) and COD (~ 25%) was observed between 72 h and 8 days of the biological treatment. The CPWW toxicity in Allium cepa seeds was lower by up to 60%, and the germination index (GI) exceeded 100% in the treated CPWW. The results of the CPWW biological treatment by bio-augmentation from native micro-organisms in the pilot-scale WTP indicated the greatest efficiency relating to the spontaneous biological treatment of CPWW. After this treatment, the discharge of effluent in the environment would not have toxic effects on the plants.

Solid coffee waste as alternative to produce carotenoids with antioxidant and antimicrobial activities.

Special coffee production involves particular sensorial characteristics of the beverage as well as the production system of coffee beans, in particular environmental issues like water and solid waste disposal. Many countries around the world have problems with that waste disposal. The possibility to integrate the commodite production with suitable agricultural practices was the focus on this work using the solid waste to produce pigments by yeasts with biological activities. The better carotenoids production was tested previously in husk and pulp extract. The production of total carotenoids by yeast was 16.36 ±â€¯0073 mg L-1 was 21.35 ±â€¯0067 mg L-1, in the pulp extract and husk extract respectively. Carotenoids produced, exhibited antioxidant and antimicrobial activities against pathogenic bacteria such as Salmonella colorless, Escherichia coli, Staphylococcus aureus and Listeria monocytogenes as well as toxigenic fungi like Aspergillus flavus, A. parasiticus, A. carbonarius and A. ochraceus. These characteristics of the pigments are important to replace the artificial ones commonly used in food and pharmaceutical industries allowing the consumers to choose more natural products at lower costs.

Volatile compounds flavoring obtained from Brazilian and Mexican spirit wastes by yeasts.

Vinasse is a waste obtained from the production of beverages, such as tequila and cachaça. The presence of acids, alcohols, sugars, minerals, amino acids, peptides, and nitrogen salts make vinasse a hazardous liquid waste to the environment, affecting the fauna, flora, and microbiota of rivers and lagoons. This study used biological treatment concomitant to volatile compound production. The yeasts used in the study were Saccharomyces cerevisiae (CCMA 0187 and CCMA 0188), Candida parapsilosis (CCMA 0544), and Pichia anomala (CCMA 0193). A higher percentage reduction in chemical and biochemical oxygen demand was observed in the tequila vinasse than in the cachaça vinasse. However, a higher production of volatile compounds was observed in the cachaça vinasse. C. parapsilosis CCMA 0544 produced the highest concentration of 2-phenylethanol (162 mg L-1). These results indicated that the environmental damage of vinasse can be reduced by treating vinasse with yeasts, and this treatment produces aroma compounds. This biological treatment has high economic potential, especially for the tequila industry.

Diversity of microbiota found in coffee processing wastewater treatment plant.

Cultivable microbiota presents in a coffee semi-dry processing wastewater treatment plant (WTP) was identified. Thirty-two operational taxonomic units (OTUs) were detected, these being 16 bacteria, 11 yeasts and 4 filamentous fungi. Bacteria dominated the microbial population (11.61 log CFU mL- 1), and presented the highest total diversity index when observed in the WTP aerobic stage (Shannon = 1.94 and Simpson = 0.81). The most frequent bacterial species were Enterobacter asburiae, Sphingobacterium griseoflavum, Chryseobacterium bovis, Serratia marcescens, Corynebacterium flavescens, Acetobacter orientalis and Acetobacter indonesiensis; these showed the largest total bacteria populations in the WTP, with approximately 10 log CFU mL- 1. Yeasts were present at 7 log CFU mL- 1 of viable cells, with Hanseniaspora uvarum, Wickerhamomyces anomalus, Torulaspora delbrueckii, Saturnispora gosingensis, and Kazachstania gamospora being the prevalent species. Filamentous fungi were found at 6 log CFU mL- 1, with Fusarium oxysporum the most populous species. The identified species have the potential to act as a biological treatment in the WTP, and the application of them for this purpose must be better studied.

Microbiological diversity associated with the spontaneous wet method of coffee fermentation.

The evaluation of the microbiota present during coffee wet fermentation was done in two distinct regions of Minas Gerais, Brazil: one farm in the South of Minas Gerais (Lavras=L) and another farm in the savannah region (Monte Carmelo=MC). The yeast population ranged from 2.48 to 4.92 log CFU/g and from 2 to 4.81 log CFU/g, the mesophilic bacteria population ranged from 3.83 to 8.47 log CFU/g and from 5.37 to 7.36 log CFU/g, and the LAB population ranged from 2.57 to 5.66 log CFU/g and from 3.40 to 4.49 log CFU/g in the L and MC farms, respectively. Meyerozyma caribbica and Hanseniaspora uvarum were the dominant yeasts in coffee wet fermentation at L farm, and Torulaspora delbrueckii was the dominant yeast at MC farm. The species Staphylococcus warneri and Erwinia persicina were the predominant bacteria at L farm, and Enterobacter asburiae and Leuconostoc mesenteroides were the dominant species at MC farm. Lactic acid was the principal acid detected, reaching 2.33 g/kg at L farm and 1.40 g/kg at MC farm by the end of the process. The volatiles composition was similar for roasted coffee from the two different regions and furans, acids, and alcohol were the main groups detected. Temporal Dominance Sensation (TDS) analyses showed that the coffee beverage from L farm was dominated by citrus and herbaceous sensory characteristics, while the coffee from MC farm was dominated by citrus, herbaceous, and nuts sensory characteristics. Evaluating the microbiota in these two regions was important in improving the knowledge of the microbial species present during coffee wet fermentation in Brazil.

Influence of different frequencies of transcutaneous electrical nerve stimulation on the threshold and pain intensity in young subjects.

OBJECTIVE: To investigate the effects of different transcutaneous electrical nerve stimulation frequencies in nociception front of a pressure pain threshold and cold in healthy individuals. METHODS: Twenty healthy subjects were divided into four groups, all of which have gone through all forms of electrical stimulation at different weeks. Assessments were pre and post-therapy, 20 and 60 minutes after stimulation. To evaluate the pressure pain threshold, an algometer was used with one tapered tip, pressing the hypothenar region until voluntary report the word "pain". Cold pain intensity was assessed by immersion in water at 5°C for 30 seconds; at the end, the subject was asked to quantify the pain intensity on a Visual Analog Scale for Pain. For electrical stimulation, two electrodes were used near the elbow, for 20 minutes, with an intensity strong, but not painful. The frequency was in accordance with the group: 0Hz (placebo); 7Hz; 100Hz; and 255Hz. RESULTS: Both for the assessment of pressure pain threshold as the cold pain intensity, there was no significant difference (p>0.05). CONCLUSION: We conclude that the use of transcutaneous electrical nerve stimulation on dermatomes C6 to C8 produced no significant change in pressure pain threshold or cold discomfort.

Influence of different frequencies of transcutaneous electrical nerve stimulation on the threshold and pain intensity in young subjects / As influências de diferentes frequências da estimulação elétrica nervosa transcutânea no limiar e intensidade de dor em indivíduos jovens

Objective To investigate the effects of different transcutaneous electrical nerve stimulation frequencies in nociception front of a pressure pain threshold and cold in healthy individuals. Methods Twenty healthy subjects were divided into four groups, all of which have gone through all forms of electrical stimulation at different weeks. Assessments were pre and post-therapy, 20 and 60 minutes after stimulation. To evaluate the pressure pain threshold, an algometer was used with one tapered tip, pressing the hypothenar region until voluntary report the word “pain”. Cold pain intensity was assessed by immersion in water at 5°C for 30 seconds; at the end, the subject was asked to quantify the pain intensity on a Visual Analog Scale for Pain. For electrical stimulation, two electrodes were used near the elbow, for 20 minutes, with an intensity strong, but not painful. The frequency was in accordance with the group: 0Hz (placebo); 7Hz; 100Hz; and 255Hz. Results Both for the assessment of pressure pain threshold as the cold pain intensity, there was no significant difference (p>0.05). Conclusion We conclude that the use of transcutaneous electrical nerve stimulation on dermatomes C6 to C8 produced no significant change in pressure pain threshold or cold discomfort. .

Objetivo Investigar os efeitos de diferentes frequências da estimulação elétrica nervosa transcutânea na nocicepção, frente a um estímulo doloroso pressórico e ao frio, em indivíduos saudáveis. Métodos Participaram 20 indivíduos saudáveis, divididos em 4 grupos, sendo que todos passaram por todas as formas de eletroestimulação, em semanas diferentes. As avaliações ocorreram nos seguintes períodos: pré-aplicação, pós-aplicação, 20 e 60 minutos após a eletroestimulação. Para avaliar o limiar de dor à pressão, foi utilizado um algômetro com ponta afilada, pressionando na região hipotenar, até o voluntário relatar a palavra “dor”. A intensidade de dor ao frio foi avaliada por meio de imersão em água a 5°C, durante 30 segundos; ao final, pediu-se para que o indivíduo quantificasse a intensidade álgica em uma Escala Visual Analógica de Dor. Para a eletroestimulação, foram utilizados dois eletrodos próximos ao cotovelo, durante 20 minutos, com intensidade referida como forte, porém não dolorosa. A frequência esteve de acordo com o grupo: 0Hz (placebo); 7Hz; 100Hz; e 255Hz. Resultados Tanto para a avaliação do limiar de dor à pressão quanto da intensidade ao frio, não houve diferença significativa (p>0,05). Conclusão O uso da estimulação elétrica nervosa transcutânea, sobre os dermátomos de C6 a C8, não produziu alteração significativa no limiar de dor à pressão e nem no desconforto ao frio. .

Inoculation of starter cultures in a semi-dry coffee (Coffea arabica) fermentation process.

The aim of this study was to evaluate the use of yeasts as starter cultures in coffee semi-dry processing. Arabica coffee was inoculated with one of the following starter cultures: Saccharomyces cerevisiae UFLA YCN727, S. cerevisiae UFLA YCN724, Candida parapsilosis UFLA YCN448 and Pichia guilliermondii UFLA YCN731. The control was not inoculated with a starter culture. Denaturing gradient gel electrophoresis (DGGE) was used to assess the microbial population, and organic acids and volatile compounds were quantified by HPLC and HS-SPME/GC, respectively. Sensory analyses were evaluated using the Temporal Dominance of Sensations (TDS). DGGE analysis showed that the inoculated yeasts were present throughout the fermentation. Other yeast species were also detected, including Debaryomyces hansenii, Cystofilobasidium ferigula and Trichosporon cavernicola. The bacterial population was diverse and was composed of the following genera: Weissella, Leuconostoc, Gluconobacter, Pseudomonas, Pantoea, Erwinia and Klebsiella. Butyric and propionic acids, were not detected in any treatment A total of 47 different volatiles compounds have been identified. The coffee inoculated with yeast had a caramel flavor that was not detected in the control, as assessed by TDS. The use of starter cultures during coffee fermentation is an interesting alternative for obtaining a beverage quality with distinctive flavor.

Efficiency of physicochemical and biological treatments of vinasse and their influence on indigenous microbiota for disposal into the environment.

Molasses-based distilleries are one of the most polluting industries generating large volumes of high strength wastewater called vinasse. Different processes covering anaerobic, aerobic as well as physicochemical methods have been employed to treat this effluent. This study evaluated the microbial communities present in the vinasse during different stages of its treatment by traditional and molecular methods. The analysis of the efficiency of each treatment was performed by physicochemical parameters and toxicity analysis. The treatment of vinasse was performed in the following steps: high flow fermentation; filtration; chemical flakes; low-flow fermentation; filtration; and neutralization. The physicochemical analysis in different stages of the vinasse treatment demonstrated that phases of treatment influenced the performance of the evaluated parameters. Among the 37 parameters, 9 were within the limits established by the Commission for Environmental Policy of Minas Gerais, Brazil (COPAM), especially BOD (96.7% of pollution reduction), suspended solids (99.9%), pH, copper (88%), iron (92.9%), and manganese (88%). Some parameters, even after treatment, did not fit the maximum allowed by legislation. The microbial population decreased reaching 3 log CFU/ml present in the steps of the flakes chemical and disinfection treatment of vinasse. Lactobacillus brevis and Pichia kudriavzevii were present in all stages of the treatments, showing that these microorganisms were resistant and demonstrated that they might be important in the treatment of vinasse. The vinasse showed a significant reduction of pollution load after the disinfection treatment however still should not be discarded into water bodies because the high values of tannins and sediment solids, but suggest the use of the effluent in the cooling coil during the distillation process of the beverage.

Evaluation of a potential starter culture for enhance quality of coffee fermentation.

The coffee fermentation is characterized by the presence of different microorganisms belonging to the groups of bacteria, fungi and yeast. The objectives of this work were to select pectinolytic microorganisms isolated from coffee fermentations and evaluate their performance on coffee pulp culture medium. The yeasts and bacteria isolates were evaluated for their activity of polygalacturonase (PG), pectin lyase (PL) and pectin methylesterase (PME) and metabolites production. Among 127 yeasts isolates and 189 bacterial isolates, 15 were pre-selected based on their ability to produce PL and organic compounds. These isolates were strains identified as Bacillus cereus, Bacillus megaterium, Bacillus subtilis, Candida parapsilosis, Pichia caribbica, Pichia guilliermondii and Saccharomyces cerevisiae. When cultivated in Coffee peel and pulp media in single culture or two by two mixed inocula, different behavior concerning to PME, PL and PG were found. The two principal components PC1 and PC2 accounted for 45.27 and 32.02 % of the total variance. UFLA CN727 and UFLA CN731 strains were grouped in the positive part of PC1 being characterized by 1,2-propanediol, hexanoic acid, decanoic acid, nonanoic acid and ethyl acetate. The UFLA CN448 and UFLA CN724 strains were grouped in the negative part of PC1 and were mainly characterized by guaiacol, butyric acid and citronellol. S. cerevisiae UFLACN727, P. guilliermondii UFLACN731 and C. parapsilosis UFLACN448 isolates are promising candidates to be tested in future studies as coffee starter cultures.

Molecular ecology and polyphasic characterization of the microbiota associated with semi-dry processed coffee (Coffea arabica L.).

This work was aimed at isolating and identifying the microbiota present during the semi-dry method of coffee processing using polyphasic methods and to evaluate microbial diversity with PCR-DGGE. Samples of Coffea arabica L. were collected during different processing stages in southern Minas Gerais, Brazil. The bacterial and fungal isolates were phenotypically characterised and grouped according to the ARDRA technique, in which the 16-23S and ITS1-5.8S regions of the rDNA were sequenced for species identification. The bacterial counts varied from 3.7 to 7 log CFU g(-1). The yeast counts ranged from 3.4 to 6.9 log CFU g(-1), and the filamentous fungal population varied from 2 to 3.7 log CFU g(-1). Bacillus subtilis, Escherichia coli, Enterobacter agglomerans, Bacillus cereus and Klebsiella pneumoniae were the predominant bacteria detected during the processing of the coffee, and Pichia anomala, Torulaspora delbrueckii and Rhodotorula mucilaginosa were the dominant yeasts. All of the yeast and bacterial species detected by PCR-DGGE were isolated using culture-dependent methods, with the exception of one uncultivable bacterial species. Aspergillus was the most common genus among the filamentous fungal isolates. The use of polyphasic methods allowed a better characterization of the microbiota that is naturally present in semi-dry processed coffee.

Microbial diversity in a bagasse-based compost prepared for the production of Agaricus brasiliensis / Diversidade microbiana em composto a base de bagaço de cana preparado para produção de Agaricus brasiliensis

Edible mushrooms are renowned for their nutritional and medicinal properties and are thus of considerable commercial importance. Mushroom production depends on the chemical composition of the basic substrates and additional supplements employed in the compost as well as on the method of composting. In order to minimise the cost of mushroom production, considerable interest has been shown in the use of agro-industrial residues in the preparation of alternative compost mixtures. However, the interaction of the natural microbiota present in agricultural residues during the composting process greatly influences the subsequent colonisation by the mushroom. The aim of the present study was to isolate and identify the microbiota present in a sugar cane bagasse and coast-cross straw compost prepared for the production of Agaricus brasilienses. Composting lasted for 14 days, during which time the substrates and additives were mixed every 2 days, and this was followed by a two-step steam pasteurisation (55 - 65ºC; 15 h each step). Bacteria, (mainly Bacillus and Paenibacillus spp. and members of the Enterobacteriaceae) were the predominant micro-organisms present throughout the composting process with an average population density of 3 x 10(8) CFU/g. Actinomycetes, and especially members of the genus Streptomyces, were well represented with a population density of 2 - 3 x 10(8) CFU/g. The filamentous fungi, however, exhibited much lower population densities and were less diverse than the other micro-organisms, although Aspergillus fumigatus was present during the whole composting process and after pasteurisation.

Os cogumelos comestíveis são apreciados pelas suas propriedades nutricionais e medicinais e, por essa razão, possuem alto valor econômico. A produção de cogumelos depende da composição química dos substratos básicos, dos suplementos utilizados e da preparação do composto no qual o fungo será cultivado. Considerando-se que os custos de produção precisam ser minimizados, os resíduos agroindustriais representam uma fonte alternativa e econômica para a preparação do composto. A interação da microbiota natural dos resíduos agrícolas durante o processo de compostagem influencia a subseqüente colonização do cogumelo. Visando-se a produção de A. brasiliensis, o presente trabalho objetivou isolar e identificar a microbiota presente no composto preparado a partir de bagaço de cana e capim coast-cross. O processo de compostagem durou 14 dias com reviragens da pilha a cada dois dias, o qual foi seguido de pasteurização (55 65 ºC) em duas fases por 15 h cada. As bactérias (principalmente Bacillus, Paenibacillus e espécies da família Enterobacteriaceae) foram os microrganismos predominantes durante todo o processo com uma densidade populacional média de 3.0 x 10(8) UFC/g. Os actinomicetos, principalmente os do gênero Streptomyces, estiveram bem representados, com uma densidade populacional de 2.0 a 3.0 x 10(8) UFC/g. Os fungos filamentosos foi a classe de microrganismos com menor densidade populacional e menor diversidade, embora a espécie Aspergillus fumigatus esteve presente durante todo o processo de compostagem e também após a pasteurização do composto.

Microbial diversity in a bagasse-based compost prepared for the production of Agaricus brasiliensis.

Edible mushrooms are renowned for their nutritional and medicinal properties and are thus of considerable commercial importance. Mushroom production depends on the chemical composition of the basic substrates and additional supplements employed in the compost as well as on the method of composting. In order to minimise the cost of mushroom production, considerable interest has been shown in the use of agro-industrial residues in the preparation of alternative compost mixtures. However, the interaction of the natural microbiota present in agricultural residues during the composting process greatly influences the subsequent colonisation by the mushroom. The aim of the present study was to isolate and identify the microbiota present in a sugar cane bagasse and coast-cross straw compost prepared for the production of Agaricus brasilienses. Composting lasted for 14 days, during which time the substrates and additives were mixed every 2 days, and this was followed by a two-step steam pasteurisation (55 - 65°C; 15 h each step). Bacteria, (mainly Bacillus and Paenibacillus spp. and members of the Enterobacteriaceae) were the predominant micro-organisms present throughout the composting process with an average population density of 3 x 10(8) CFU/g. Actinomycetes, and especially members of the genus Streptomyces, were well represented with a population density of 2 - 3 x 10(8) CFU/g. The filamentous fungi, however, exhibited much lower population densities and were less diverse than the other micro-organisms, although Aspergillus fumigatus was present during the whole composting process and after pasteurisation.

Succession of bacterial and fungal communities during natural coffee (Coffea arabica) fermentation.

Bacteria, yeasts and filamentous fungi were isolated during natural coffee processing. Bacteria were isolated in greater numbers at the beginning of the fermentation, when the moisture of the coffee beans was around 68%. Gram-positive bacteria represented 85.5% of all bacteria isolated, and Bacillus was the predominant genus (51%). Gram-negative species of the genera Serratia, Enterobacter and Acinetobacter were also found. Approximately 22% of 940 randomly chosen isolates of microorganisms were yeasts. Debaryomyces (27%), Pichia (18.9%) and Candida (8.0%) were the most commonly found genera, and these three genera tended to appear more often as the fruit was fermented and dried. Aspergillus was the most abundant genus besides Penicillium, Fusarium and Cladosporium, with 42.6% of the total fungi isolates. The genera and species identified included members known to have pectinase and cellulase activities. Of the 10 organic acids analyzed and quantified in coffee beans, acetic and lactic acids may have been generated by microbial activity. Butyric acid was not detected in any sample.

Incidence and distribution of filamentous fungi during fermentation, drying and storage of coffee (Coffea arabica L. ) beans / Incidência e distribuição de fungos filamentosos durante a fermentação, secagem e armazenamento de frutos e grãos de café (Coffea arabica L. )

The objective of this work was to isolate and characterize filamentous fungi present in different stages of harvest, fermentation, drying and storage of coffee beans processed by natural method. The cherries were hand-picked and then placed on a cement drying platform where they remained until reached 11 percent of humidity. Microbial counts were found in all samples during fermentation and drying of the coffee beans. Counts of fungi in the coffee cherries collected from the tree (time 0) were around 1.5 x 10³ CFU/g. This number increased slowly during the fermentation and drying reaching values of 2 x 10(5) CFU/g within 22 days of processing. Two hundred and sixty three isolates of filamentous fungi were identified. The distribution of species during fermentation and drying was very varied while there was a predominance of Aspergillus species during storage period. The genera found were Pestalotia (4), Paecelomyces (4), Cladosporium (26), Fusarium (34), Penicillium (81) and Aspergillus (112) and comprised 38 different species.

O objetivo deste estudo foi isolar e caracterizar fungos filamentosos presentes em diferentes estágios de beneficiamento de café processado pelo método natural, incluindo: colheita, fermentação, secagem e armazenamento. O café cereja foi colhido manualmente e então colocado em uma plataforma de cimento, onde permaneceu até atingir 11 por cento de umidade. A contagem microbiana foi realizada em todas as amostras durante a fermentação e secagem do café. A população de fungos filamentosos no café cereja ainda nos pés (tempo 0) foi em torno de 1,5 x 10³ UFC/g. Este número aumentou vagarosamente durante a fermentação e secagem, alcançando valores de 2 x 10(5) UFC/g em 22 dias do processamento. Duzentos e sessenta e três isolados de fungos filamentosos foram identificados. A distribuição das espécies durante fermentação e secagem foi bastante variada, mas no armazenamento dos grãos ocorreu o predomínio de espécies de Aspergillus. Foram encontradas 38 espécies de fungos distribuídas nos seguintes gêneros: Pestalotia (4), Paecelomyces (4), Cladosporium (26), Fusarium (34), Penicillium (81) e Aspergillus (112).