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Braz. j. microbiol ; 48(3): 592-601, July-Sept. 2017. tab, graf
Article in English | LILACS | ID: biblio-889150


Abstract The aim of this study was to develop a kefir apple-based vinegar and evaluate this fermentation process using new methodology with Biospeckle Laser. Brazilian kefir grains were inoculated in apple must for vinegar production. In this study, the microbial community present in kefir, and correspondent vinegar, was investigated using Matrix Assisted Laser Desorption/Ionization - Time of Flight Mass Spectrometry (MALDI-TOF MS) technique. Saccharomyces cerevisiae, Lactobacillus paracasei, Lactobacillus plantarum, Acetobacter pasteurianus and Acetobacter syzygii were the microbial species identified. S. cerevisiae, L. plantarum, A. pasteurianus and A. syzygii were found in smaller quantities at the beginning of the alcoholic fermentation, but were found throughout the alcoholic and acetic fermentation. Kefir grains were able to utilize apple must as substrate to produce ethanol, and acetic acid. Acetate, volatile alcohols and aldehydes in the vinegar-based kefir were also produced. The yield of acetic acid in the kefir vinegars was ∼79%. The acetic acid concentration was ∼41 g L-1, reaching the required standard for the Brazilian legislation accepts it as vinegar (4.0% acetic acid). Kefir vinegar showed good acceptance in the sensory analysis. The technology proposed here is novel by the application of immobilized-cell biomass (kefir grains) providing a mixed inocula and eliminating the use of centrifuge at the end of the fermentative process. This step will save energy demand and investment. This is the first study to produce apple vinegar using kefir grains.

Humans , Alcoholic Beverages/microbiology , Kefir/analysis , Malus/microbiology , Acetic Acid/analysis , Acetic Acid/metabolism , Acetobacter/isolation & purification , Acetobacter/metabolism , Biodiversity , Brazil , Ethanol/analysis , Ethanol/metabolism , Fermentation , Food Handling , Kefir/microbiology , Lactobacillus/isolation & purification , Lactobacillus/metabolism , Malus/metabolism , Saccharomyces cerevisiae/isolation & purification , Saccharomyces cerevisiae/metabolism , Taste
Braz. j. microbiol ; 46(4): 1207-1216, Oct.-Dec. 2015. tab, graf
Article in English | LILACS | ID: lil-769650


Bacaba chicha is a beverage prepared by the indigenous Umutina people from the bacaba fruit (Oenocarpus bacaba), a purple berry that is rich in fat and carbohydrates, as well as a source of phenolic compounds. In this study, samples of bacaba chicha beverage were collected, and the microbial community was assessed using culture-dependent and -independent techniques. The nutritional composition and metabolite profiles were analyzed, and species belonging to lactic acid bacteria (LAB) and yeasts were detected. The LAB group detected by culture-dependent analysis included Enterococcus hormaechei and Leuconostoc lactis. Polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) detected additional Propionibacterium avidum, Acetobacter spp., and uncultured bacteria. Pichia caribbica and Pichia guilliermondii were detected in a culture-dependent method, and Pichia caribbica was confirmed by PCR-DGGE analysis. The pH value of the beverage was 6.2. The nutritional composition was as follows: 16.47 ± 0.73 g 100 mL-1 dry matter, 2.2 ± 0.0 g 100 mL-1 fat, 3.36 ± 0.44 g 100 mL-1 protein, and 10.87 ± 0.26 g 100 mL-1 carbohydrate. The metabolites detected were 2.69 g L-1 succinic acid, 0.9 g L-1 acetic acid, 0.49 g L-1 citric acid, 0.52 g L-1 ethanol, and 0.4 g L-1 glycerol. This is the first study to identify microbial diversity in bacaba chicha spontaneous fermentation. This study is also the starting step in the immaterial record of this Brazilian indigenous beverage prepared from bacaba fruit.

Humans , Chronic Disease/economics , Health Care Costs/statistics & numerical data , Health Expenditures/statistics & numerical data , Models, Econometric , State Government , Absenteeism , Centers for Disease Control and Prevention, U.S. , Cost of Illness , International Classification of Diseases , Medicaid/economics , Medicare/economics , Regression Analysis , United States
Biosci. j. (Online) ; 29(5-Supplement 1): 1678-1686, nov. 2013. tab, ilus
Article in English | LILACS | ID: biblio-967403


Thirty-two strains of Lactobacillus plantarum UFLA SAU from pork sausages, pre-selected for some features for probiotic application, were utilized in this study to evaluate their adhesive properties and compare the results against the three pathogens also tested. Strains were tested for autoaggregation and coaggregation capacity and Microbial Adhesion To Solvents (MATS) at the time intervals of 0, 1, 2, 3 and 4 h. Our findings revealed that UFLA SAU strains have a high autoaggregative capacity and coaggregative ability with pathogens, especially Listeria monocytogenes. In relation to adhesion to solvents, in general, L. plantarum strains showed hydrophilic cell surface properties and an important electron donor and basic character. Adhesive properties were markedly separated for the strains under study by Principal Component Analysis software. UFLA SAU 132, 226 and 87 were differentiated by autoaggregation ability. UFLA SAU 11 and Listeria monocytogenes were characterized by adhesion to solvents. UFLA SAU 14, 18 and 172 showed high coaggregation with Escherichia coli, Salmonella Typhi and Listeria monocytogenes. In comparison to the pathogens tested, many UFLA SAU strains presented higher adhesive capacity. These tests should be used for screening and identifying potentially adherent microorganisms. Adhesive properties are important features for the choice of probiotic strains and confer various applications, such as in the pharmaceutical (therapeutic or prophylactic) and food (functional foods) industries.

Trinta e duas estirpes de linguiça suína, Lactobacillus plantarum UFLA SAU, pré-selecionadas com algumas características para aplicação probiótica, foram utilizadas neste estudo para avaliar suas propriedades adesivas e comparar os resultados com três patógenos também testados. As estirpes foram testadas para autoagregação, coagregação e capacidade de adesão microbiana aos solventes (MATS) nos tempos de 0, 1, 2, 3 e 4 h. Nossos resultados revelaram que estirpes UFLA SAU apresentam alta capacidade autoagregativa e coagregativa com patógenos, especialmente com Listeria monocytogenes. Em relação à adesão aos solventes, de um modo geral, as estirpes de L. plantarum mostraram propriedades hidrofílicas de superfície celular e um importante caráter básico e elétron doador. Propriedades adesivas foram marcadamente separadas para as estirpes em estudo através da Análise de Componentes Principais. UFLA SAU 132, 226 e 87 foram diferenciadas pela capacidade de autoagregação. UFLA SAU 11 e Listeria monocytogenes foram caracterizadas por adesão aos solventes. UFLA SAU 14, 18 e 172 apresentaram coagregação com Escherichia coli, Salmonella Typhi e Listeria monocytogenes. Em comparação aos patógenos testados, muitas estirpes UFLA SAU apresentaram maior capacidade adesiva. Estes testes podem ser úteis para a triagem e identificação de micro-organismos potencialmente aderentes. Propriedades adesivas são importantes características para a escolha de estirpes probióticas e conferem várias aplicações, tais como nas indústrias: farmacêutica (terapêutico ou profilático) e de alimentos (alimentos funcionais).

Probiotics , Lactobacillus plantarum , Noxae
Braz. j. microbiol ; 44(3): 935-944, July-Sept. 2013. ilus, graf, tab
Article in English | LILACS | ID: lil-699788


Sixty six indigenous Saccharomyces cerevisiae strains were evaluated in stressful conditions (temperature, osmolarity, sulphite and ethanol tolerance) and also ability to flocculate. Eighteen strains showed tolerant characteristics to these stressful conditions, growing at 42 ºC, in 0.04% sulphite, 1 mol L-1 NaCl and 12% ethanol. No flocculent characteristics were observed. These strains were evaluated according to their fermentative performance in sugar cane juice. The conversion factors of substrates into ethanol (Yp/s), glycerol (Yg/s) and acetic acid (Yac/s), were calculated. The highest values of Yp/s in sugar cane juice fermentation were obtained by four strains, one isolated from fruit (0.46) and the others from sugar cane (0.45, 0.44 and 0.43). These values were higher than the value obtained using traditional yeast (0.38) currently employed in the Brazilian bioethanol industry. The parameters Yg/s and Yac/s were low for all strains. The UFLA FW221 presented the higher values for parameter related to bioethanol production. Thus, it was tested in co-culture with Lactobacillus fermentum. Besides this, a 20-L vessel for five consecutive batches of fermentation was performed. This strain was genetically stable and remained viable during all batches, producing high amounts of ethanol. The UFLA FW221 isolated from fruit was suitable to produce bioethanol in sugar cane juice. Therefore, the study of the biodiversity of yeasts from different environmental can reveal strains with desired characteristics to industrial applications.

Stress, Physiological , Saccharomyces cerevisiae/physiology , Acetic Acid/metabolism , Brazil , Carbohydrate Metabolism , Cell Aggregation , Ethanol/metabolism , Ethanol/toxicity , Fermentation , Glycerol/metabolism , Saccharomyces cerevisiae/drug effects , Saccharomyces cerevisiae/growth & development , Saccharomyces cerevisiae/radiation effects , Sodium Chloride/metabolism , Sodium Chloride/toxicity , Sulfites/metabolism , Sulfites/toxicity , Temperature
Braz. j. microbiol ; 42(2): 693-702, Apr.-June 2011. ilus, graf, tab
Article in English | LILACS-Express | LILACS | ID: lil-590016


Microbial ecology and chemical composition of Brazilian kefir beverage was performed. The microorganisms associated with Brazilian kefir were investigated using a combination of phenotypic and genotypic methods. A total of 359 microbial isolates were identified. Lactic acid bacteria (60.5 percent) were the major isolated group identified, followed by yeasts (30.6 percent) and acetic acid bacteria (8.9 percent). Lactobacillus paracasei (89 isolates), Lactobacillus parabuchneri (41 isolates), Lactobacillus casei (32 isolates), Lactobacillus kefiri (31 isolates), Lactococcus lactis (24 isolates), Acetobacter lovaniensis (32 isolates), Kluyveromyces lactis (31 isolates), Kazachstania aerobia (23 isolates), Saccharomyces cerevisiae (41 isolates) and Lachancea meyersii (15 isolates) were the microbial species isolated. Scanning electron microscopy showed that the microbiota was dominated by bacilli (short and curved long) cells growing in close association with lemon-shaped yeasts cells. During the 24 h of fermentation, the protein content increased, while lactose and fat content decreased. The concentration of lactic acid ranged from 1.4 to 17.4 mg/ml, and that of acetic acid increased from 2.1 to 2.73 mg/ml. The production of ethanol was limited, reaching a final mean value of 0.5 mg/ml.

Acta sci., Biol. sci ; 33(1): 93-97, Jan.- Mar. 2011. ilus
Article in English | LILACS | ID: biblio-875724


The nutritional requirements of A. brasiliensis in culture media were assessed by supplementing a basal medium (g L-1): (glucose, 10, KH2PO4, 1, MgSO4.7H2O, 0.5, [NH4]2SO4, 1, pH 5.5) with CaCl2, trace elements (FeSO4.7H2O; MnCl2.4H2O; ZnSO4.7H2O; CuSO4.5H2O), casein, yeast extract, peptone, B-vitamins or amino acids. Evaluations were based on the mycelial growth in solid or liquid culture (mm day-1 or mg day-1) and visual analysis of the colony. The addition of CaCl2 and trace elements was very important for the major mycelial growth of the fungi. The addition of casein and inositol to the medium did not have a significant effect on growth. The best growth result in solid medium was obtained with the basal medium plus the addition of yeast extract and peptone. In relation to the other nutrient sources, the mycelial growth in the presence of amino acids darkened the medium after two weeks. The addition of B-vitamins to the basal medium lead to slower mycelial growth; however, growth was more visually dense when compared to other nutritional sources. B-vitamins added separately did not have the same result, suggesting that the fungus requires two or more vitamins at the same time for better mycelial growth.

Os requerimentos nutricionais de A. brasiliensis foram avaliados, com a suplementação de um meio basal (g L-1): (glicose, 10, KH2PO4, 1, MgSO4.7H2O, 0.5, [NH4]2SO4, 1, pH 5.5) com CaCl2, micronutrientes (FeSO4.7H2O; MnCl2.4H2O; ZnSO4.7H2O; CuSO4.5H2O), caseína, extrato de levedura, peptona, vitaminas do complexo B ou aminoácidos. O crescimento micelial foi avaliado em meio sólido e líquido, considerando velocidade de crescimento e produção de massa micelial (mm dia-1 ou mg dia-1) e análise visual da colônia. A adição de CaCl2 e micronutrientes foi muito importante para o melhor crescimento micelial do fungo, enquanto que a adição de caseina e inositol não apresentou efeito significativo sobre o crescimento. O melhor crescimento em meio sólido foi obtido quando o meio basal foi suplementado com extrato de levedura e peptona. Quando o fungo foi cultivado no meio basal suplementado com aminoácidos, observou-se um escurecimento do meio após duas semanas de cultivo. A adição de vitaminas proporcionou um crescimento micelial mais lento no meio sólido, entretanto, mais denso em relação ao meio suplementado com outros nutrientes. Quando as vitaminas do complexo B foram adicionadas separadamente não se observou o mesmo resultado, o que sugere que o fungo requer duas ou mais vitaminas no meio para melhorar o crescimento micelial.

Agaricales , Amino Acids , Trace Elements , Vitamin B Deficiency
Braz. j. microbiol ; 40(3): 590-600, Sept. 2009.
Article in English | LILACS-Express | LILACS | ID: lil-522480


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.

Braz. j. microbiol ; 39(3): 521-526, July-Sept. 2008. tab
Article in English | LILACS | ID: lil-494544


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).

Coffea Cruda/analysis , Coffea Cruda/toxicity , Food Handling , Fungi/isolation & purification , In Vitro Techniques , Coffee , Fermentation , Food Samples
Braz. j. microbiol ; 37(4): 499-504, Oct.-Dec. 2006. tab
Article in English | LILACS | ID: lil-442201


Sugar cane silage has a potential for animal feeding, but uncontrolled growth of undesirable microorganisms may cause nutritional losses and affect the animal productivity and health. The objective of this work was to evaluate the microbiological quality and chemical composition of ensiled sugar cane with and without nutritional additives after 30 days of fermentation. Yeasts, filamentous fungi and distinct groups of bacteria were enumerated by plate count methods and the chemical analyzes comprised dry matter, crude protein, fiber content, lignin, and pH. Facultative aerobic bacteria and filamentous fungi were not detected during the fermentative process in any of the treatments. The number of yeasts in five varieties of sugar cane silage without additives was about 6.55 Log CFU g-1 of silage, and with 1 percent ammonium sulfate and 1 percent urea were about 5.86 and 5.50 Log CFU g-1 of silage, respectively. The lactic acid bacteria (LAB) count without additive was about 8.62 Log CFU g-1 of silage, and with 1 percent ammonium sulfate and 1 percent urea the count was about 6.40 and 6.54 Log CFU g-1 of silage, respectively. The average percent of dry material in the three treatments was 20.76 percent. The addition of ammonium sulphate and urea has decreased the microbial load after 30 days but it has increased the total crude protein concentration. Additives also affected neutral detergent fiber, acid detergent fiber and lignin content in all five varieties of sugar cane silage.

A silagem de cana-de-açúcar apresenta grande potencial para o uso na alimentação animal, entretanto o crescimento de microrganismos não desejáveis durante o processo fermentativo pode causar perdas nutricionais e conseqüentemente afetar o rendimento de produção e também a saúde dos animais. Este estudo objetivou avaliar a qualidade microbiológica e a composição química de silagens de cana-de-açúcar em silos experimentais com e sem a adição de aditivos nutritivos durante o período de 30 dias. Bactérias aeróbicas facultativas e fungos filamentosos não foram detectados nas amostras em nenhum dos tratamentos analisados. A população de leveduras nas silagens das cinco variedades de cana-de-açúcar sem aplicação de aditivos foi em média 6,55 log UFC g-1 de silagem e, com aplicação de 1 por cento de sulfato de amônia e 1 por cento de uréia foi em média de 5,86 e 5,50 log UFC g-1 de silagem, respectivamente. A população de bactérias do ácido lático nos silos sem aditivos foi de 8,62 log UFC g-1 de silagem e nos silos com sulfato de amônio e uréia foi de 6,40 e 6,54 log UFC g-1 de silagem, respectivamente. A percentagem média de matéria seca das silagens nos três tratamentos foi de 20,76 por cento. A adição dos nutrientes nitrogenados diminuiu a população microbiana após os 30 dias da cana de açúcar ensilada, mas aumentou a concentração de proteína bruta. A presença dos aditivos também afetou a concentração das fibras de detergente neutro e ácida e lignina nas cinco variedades de cana de açúcar ensiladas.

Bacteria , Food Additives , In Vitro Techniques , Lactic Acid , Saccharum , Silage , Culture Media , Fermentation
Braz. j. microbiol ; 32(2): 117-122, Apr.-Jun. 2001. tab
Article in English | LILACS | ID: lil-391991


Setenta e duas embalagens de iogurtes de quatro indústrias diferentes foram analisadas durante três épocas diferentes com intervalo mensal. A população microbiana total encontrada foi em torno de 6 x 107 células g-1 de iogurte. A contagem de leveduras variou entre 1 a 2.700 células g-1. Não foi possível observar uma sistemática contaminação, mas este estudo longitudinal revelou que contaminação ad hoc e armazenamento impróprio pode levar a elevadas populações de leveduras. De modo geral foi detectada uma contaminação maior nos meses mais quentes do ano mas em valores inferiores aos encontrados em outros países. Um total de 577 isolados de leveduras foram identificados como pertencentes a 10 espécies. As leveduras mais abundantes foram, em ordem, Debaryomyces hansenii, Saccharomyces cerevisiae, Mrakia frigida, Hansenula spp., Candida parapsilosis, Debaryomyces castellii e Candida maltosa. A levedura psicrófila, Mrakia frigida foi pela primeira vez mencionada como isolada a partir de iogurtes. Foi encontrada em algumas amostras uma pequena contaminação por espécies de Monilia e Penicillium. Os testes utilizados para crescimento sugeriram que habilidade para fermentar sacarose, crescimento a 5ºC e na presença de 300 µg g-1 de sorbato foram as três propriedades fisiológicas mais importantes para a presença destas leveduras em iogurtes. Os dados também sugerem que clima mais quente e refrigeração inadequada são as principais causas de alta nível de contaminação, aumento da diversidade e mudança na microbiota presente.

Clinical Enzyme Tests , Food Contamination/analysis , Fungi , In Vitro Techniques , Yeasts , Yogurt , Culture Media , Fermentation , Methods