Revealing the microbial diversity and physicochemical characteristics of Brazilian untreated green table olives.

AIM: This work evaluated the microbial diversity and physicochemical characteristics of fresh and fermented fruits from Brazilian untreated green table olives of the Ascolano and Grappolo cultivars. METHODS AND RESULTS: Twenty species of mesophilic bacteria, seven lactic acid bacteria, and fourteen yeast were identified. Some species prevailed over others, such as the bacteria Levilactobacillus brevis, Lacticaseibacillus paracasei subsp. paracasei, Pantoea agglomerans, Staphylococcus warneri, Bacillus simplex, B. thuringiensis, and the yeasts Candida parapsilosis, Ca. orthopsilosis, and Cryptococcus flavescen. In the olive fruit and olive brine, the sugars: sucrose, glucose, mannitol, and fructose, and the acids: acetic, citric, lactic, malic, and succinic were identified. Thirty-seven volatile compounds belonging to different chemical classes of acids, alcohols, aldehydes, esters, hydrocarbons, phenols, ketones, and ether were identified in the fruits and brine olives. CONCLUSION: The polyphasic methodology using matrix assisted laser desorption/ionization-time of flight and 16S rRNA sequencing was efficiently performed to identify microorganisms; chemical analysis helped to understand the fermentation process of olives.

Association of the essential oil of Cymbopogon citratus (DC) Stapf with nystatin against oral cavity yeasts.

Oral thrush or candidiasis is a fairly common infectious fungal disease that is mainly caused by species of the genus Candida spp. In this work, we evaluated the antifungal activity of the essential oil of Cymbopogon citratus (DC) Stapf (lemongrass) and its combination with nystatin against oral cavity yeasts. The oil was extracted by the steam distillation method, and its constituents were quantified. The yeasts were identified using MALDI-TOF MS. The sensitivity to the essential oil and its association with nystatin was verified by the minimum inhibitory concentration (MIC) and checkerboard methods. In the essential oil samples, there were 84.53% of citral. A total of 64.77% of strains were identified as Candida albicans. On susceptibility tests, 83.55% of the yeast isolates were inhibited in concentrations of ≥16 µg/mL of nystatin. C. citratus's oil was capable of inhibiting and killing all the isolates tested with concentrations that varied from 0.137 to 2.2 mg/mL. The association of oil and nystatin had an additive effect on more than 78% of the strains. The association of this herbal drug with nystatin potentialized the antifungal effect on yeast samples isolated from the oral cavity of oncologic patients.

Effect of Brazilian green propolis on microorganism contaminants of surface of Gorgonzola-type cheese.

Blue cheeses are susceptible to yeast and bacterial growth on their surface, which causes spoilage during ripening process and the formation of slime. The dairy industry frequently control the proliferation of undesirable microorganisms with natamycin and high salt concentration. The green propolis is a complex of substances that presents antimicrobial properties with great potential as preservative in the food industry. The aims of the present study were to identify the mesophilic aerobic microorganisms present on the surface of Gorgonzola-type cheese, evaluate the antifungal and antibacterial effects of the ethanol extract of green propolis (EEP) on the development of those microorganisms and verify the effects of EEP on the sensory quality of cheese. Ten yeast species belonging to genera Yarrowia, Candida, Debaryomyces and Saccharomyces were identified, as well as seven species of bacteria belonging to genera Staphylococcus, Bacillus, Enterococcus, Corynebacterium and Proteus. The EEP showed minimum biocide concentration (MBC), between 0.3% (weight/weight) and 5% for Bacillus cereus and Proteus vulgaris, respectively. Saccharomyces cerevisiae was the most sensitive species (MBC of 0.63%) and Candida parapsilosis the most resistant one (MBC of 5%). In the sensory analysis, the cheeses involved with EEP at 5% concentration did not differ from the control, while at 10%, there was a slight decrease in acceptance. The EEP has potential and feasibility to be used in Gorgonzola-type cheese, inhibiting the main bacteria and yeasts without affecting largely the sensory characteristics of the product.

Use of lignocellulose biomass for endoxylanase production by Streptomyces termitum.

Actinobacteria isolates from Brazilian Cerrado soil were evaluated for their ability to produce enzymes of the cellulolytic and xylanolytic complex using lignocellulose residual biomass. Preliminary semiquantitative tests, made in Petri plates containing carboxymethylcellulose and beechwood xylan, indicated 11 potential species producing enzymes, all belonging to the genus Streptomyces. The species were subsequently grown in pure substrates in submerged fermentation and analyzed for the production of enzymes endoglucanase, ß-glucosidase, endoxylanase, and ß-xylosidase. The best results were obtained for endoxylanase enzyme production with Streptomyces termitum(UFLA CES 93). The strain was grown on lignocellulose biomass (bagasse, straw sugarcane, and cocoa pod husk) that was used in natura or acid pretreated. The medium containing sugarcane bagasse in natura favored the production of the endoxylanase that was subsequently optimized through an experimental model. The highest enzyme production 0.387 U mL-1, (25.8 times higher), compared to the lowest value obtained in one of the trials, was observed when combining 2.75% sugar cane bagasse and 1.0 g L-1 of yeast extract to the alkaline medium (pH 9.7). This is the first study using S. termitum as a producer of endoxylanase.

Yeast Diversity in Honey and Pollen Samples from Stingless Bees in the State of Bahia, Brazil: Use of the MALDI-TOF MS/Genbank Proteomic Technique.

(1) Background: The identification of microorganisms includes traditional biochemical methods, molecular biology methods evaluating the conserved regions of rRNA, and the molecular biology of proteins (proteomics), such as MALDI-TOF MS mass spectrometry. This work aimed to identify the biodiversity of yeasts associated with stingless bee species' honey and pollen, Melipona scutellaris, Nannotrigona testaceicornes, and Tetragonisca angustula, from the region of São Gonçalo dos Campos-Bahia (BA) state, Brazil. (2) Methods: Cellular proteins were extracted from 2837 microbial isolates (pollen and honey) and identified via MALDI-TOF MS. The identified yeast species were also compared to the mass spectra of taxonomically well-characterized reference strains, available from the National Center of Biotechnology Information (NCBI) database. (3) Results: Nine yeast species were identified: Candida maltosa, Candida norvegica, Kazachstania telluris, Schizosaccharomyces pombe, Scheffersomyces insectosus, Meyerozyma guilliermondii, Brettanomyces bruxellensis, Kazachstania exigua, and Starmerella lactis-condensi. Nannotrigona testaceicornes pollen had the highest number of yeast colonies. The yeasts Brettanomyces bruxellensis and Kazachstania telluris showed high populations in the samples of Nannotrigona testaceicornes and Melipona scutellaris, respectively. This work shows that there is some sharing of the same species of yeast between honey and pollen from the same beehive. (4) Conclusions: A total of 71.84% of the identified species present a high level of confidence at the species level. Eight yeast species (Candida maltosa, Candida norvegica, Kazachstania telluris, Schizosaccharomyces pombe, Scheffersomyces insectosus, Meyerozyma guilliermondii, Kazachstania exigua, and Starmerella lactis-condensi) were found for the first time in the samples that the authors inspected. This contributes to the construction of new knowledge about the diversity of yeasts associated with stingless bee products, as well as to the possibility of the biotechnological application of some yeast species.

Novel yeasts with potential probiotic characteristics isolated from the endogenous ferment of artisanal Minas cheese.

Artisanal Minas cheese (QMA) is traditionally elaborate using raw milk and endogenous ferment (pingo - whey or rala - grated ripened cheese). In the present study, 91 yeast strains were isolated and identified from pingo and rala. Eight yeast species were identified by the MALDI-TOF mass spectrometry and confirmed by sequencing of the ITS region. The yeasts' protease and lipase activities were evaluated in addition to probiotic properties such as tolerance to low pH and bile salts, hydrophobicity, autoaggregation, co-aggregation with pathogens, and antimicrobial susceptibility. The rala ferment showed a greater variety of species. Yarrowia lipolytica was the dominant specie (52.7% of isolates), followed by the Kluyveromyces lactis and Kodamaea ohmeri (9.9 and 6.6%, respectively). From the total yeasts evaluated, 74 strains showed positive enzymatic activity: 52 strains showed lipolytic (51 Y. lipolytica and one Trichosporon japonicum) and 44 proteolytic activities (18 Y. lipolytica, 13 K. ohmeri, 11 K. lactis, and 2 Wickerhamiella sp.). All evaluated isolates demonstrated tolerance to pH 2.0, and 69 isolates supported the presence of bile salts. From them, 12 isolates showed the capacity of autoaggregation (> 30%) and hydrophobicity (> 90.0%) and were then selected for co-aggregation and antibiotic resistance assays. All selected isolates showed co-aggregation with Salmonella Enteritidis, Escherichia coli, and Listeria monocytogenes greater than 30%. None of the yeast showed sensibility to the evaluated antibiotics and antagonistic activity against the evaluated pathogens. The results demonstrated that pingo and rala have different yeast composition with different enzymatic activity, which may affect the characteristics of the cheese. Furthermore, some yeast strains: Y. lipolytica (9 strains isolated from rala) and K. ohmeri (3 strains isolated from pingo) demonstrated attractive probiotic potential.

Acclimatization of Musa spp. seedlings using endophytic Bacillus spp. and Buttiauxella agrestis strains.

The association of different species of endophytic bacteria with the rhizosphere of the host plants can stimulate growth, development and acclimatization, offering a greater quantity of seedlings, in addition to reducing the cycle, providing economic return to the producer. The objective of this study was to evaluate the effect of introduction four bacterial isolates through inoculation into the root system in three banana cultivars (Prata Anã, Grande Naine and BRS Princesa) in the acclimatization phase. The evaluated treatments were: control (nutrient broth without bacteria); Bacillus cereus strain 1 (BC1); Bacillus cereus strain 2 (BC2); Bacillus thuringiensis (BT); Buttiauxella agrestis (BA). The morphological characteristics related to the development of the plants (total height and pseudostem diameter) were evaluated throughout the acclimatization period. After 90 days of transplanting and acclimatization, root length, leaf number, dry root weight, pseudostem and leaf, leaf area, internal carbon concentration, stomatal conductance, photosynthesis rate, transpiration rate, leaf temperature and chlorophyll were evaluated. The bacteria showed different results in relation to the studied cultivars. Considering the morphological and physiological characteristics observed in this study, B. thuringiensis for the cultivars Prata Anã and Grande Naine and the B. agrestis for the cultivar BRS Princesa are recommended for the process of acclimatization of banana seedlings, as they stimulated growth of the plant, increasing the dry mass, besides promoting the growth of roots. In this way, they improved the physiological aspects of the plants and reduced the period of acclimatization of the banana.

Eco-friendly biosurfactant from Wickerhamomyces anomalus CCMA 0358 as larvicidal and antimicrobial.

Kitchen waste oil (KWO) was evaluated as a substrate for production of biosurfactant by Wickerhamomyces anomalus CCMA 0358 and was tested against Aedes aegypti larvae, the mosquito causing neglected diseases, such as dengue fever, Zika, and Chikungunya, achieving 100 % mortality in the lowest concentration (6.25 %) evaluated in 24 h. Furthermore, possible applications of this compound were evaluated as antibacterial, antiadhesive, and antifungal. At a concentration of 50 %, the biosurfactant was found to inhibit the growth of Bacillus cereus, showing high inhibitions levels against Salmonella Enteritidis, Staphylococcus aureus, and Escherichia coli. The antifungal activity was evaluated against Aspergillus, Cercospora, Colletotrichum, and Fusarium, obtaining results of up to 95 % inhibition. In addition to these promising results, the yeast W. anomalus produced the biosurfactant from an inexpensive substrate, which increases the possibility of its application in several industries owing to the low cost involved.

Prebiotic potential of pulp and kernel cake from Jerivá (Syagrus romanzoffiana) and Macaúba palm fruits (Acrocomia aculeata).

The jerivá (Syagrus romanzoffiana) and the macaúba (Acrocomia aculeata) are palm trees of the Arecaceae family, widely distributed in tropical and subtropical areas of Latin America, which have a low production cost and high productivity throughout the year. Due to the high content of lipids, their fruits have been used for oil extraction, which generates byproducts such as the pulps and the kernel cakes, a nutritionally rich byproduct that can be added into human food and, may have prebiotic potential. Therefore, the objective of this work was to characterize and evaluate the prebiotic potential of jerivá pulp (JP), macaúba pulp (MP), jerivá kernel cake (JC) and macaúba kernel cake (MC). For this, the fruits characterization was carried out through proximate composition, phenolic compounds content, and antioxidant activity, besides evaluating the antimicrobial and fermentative capacity of Bifidobacterium lactis, Lactobacillus casei, and Lactobacillus acidophilus against Escherichia coli. Jerivá and macaúba pulps and kernel cakes presented high levels of dietary fiber (20.45% JP, 37.87% JC, 19.95% MP and 35.81% MC) and high antioxidant activity, especially JP, which also showed the high values found for ABTS and DPPH (2498.49 µMTrolox·g-1 fruit and 96.97 g fruit·g-1 DPPH, respectively), has a high total phenolic content (850.62 mg GAE·100 g-1). Also, JP promoted a better growth of probiotic strains and a more relevant pH reduction when compared to the commercial prebiotic FOS. However, MP, JC, and MC were also able to favor the growth of the strains. Probiotic microorganisms were able to use JP, MP, JC, and MC and produced short-chain fatty acids such as lactic, propionic, butyric, and acetic acid, capable of promoting health benefits. Therefore, the byproducts from jerivá and macaúba oil extraction have characteristics that indicate their prebiotic potential, and maybe interesting components to increase the nutritional value of foods.

Caracterização físico-química de fermentado alcoólico de jabuticaba (Myrciaria jaboticaba Berg) / Physicochemical characterization of jabuticaba (Myrciaria jaboticaba Berg) wines

O trabalho foi realizado com o objetivo de elaborar bebida alcoólica fermentada a partir da polpa de jabuticaba e determinar a atividade antioxidante desse fermentado alcoólico. A partir da polpa, foi preparado mosto e inoculado com Saccharomyces cerevisiae, CCMA 0200, o processo fermentativo foi conduzido em fermentadores com capacidade de 6 L a22˚C. As fermentações foram monitoradas diariamente e amostras foram retiradas para análises cromatográficas. Ao fim da fermentação foram analisados o teor de compostos fenólicos totais e atividade antioxidante. O teor de etanol, ao final do processo fermentativo foi de 67,3 g.L-1 e foram identificados 25 compostos voláteis. O fermentado alcoólico de jabuticaba além de apresentar compostos importantes para o sabor e o aroma da bebida, possuem antioxidantes que são fontes de substâncias bioativas.

Caracterização fenotípica pela técnica de MALDI-TOF de Staphylococcus coagulase negativa isolados de mastite bovina em rebanhos leiteiros do Sul de Minas Gerais / Phenotypic characterization by the MALDI-TOF technique of Staphylococcus coagulase negative isolated from bovine mastitis in dairy herds of the South of Minas Gerais

O objetivo deste trabalho foi identificar espécies de Staphylococcus coagulase negativa em leite de vacas leiteiras com mastite por testes fenotípicos e pela técnica de MALDI-TOF MS. Foram utilizadas 85 isolados de estafilococos provenientes de leite de vacas com mastite. Todos os isolados foram caracterizados fenotipicamente por avaliação da morfologia das colônias, coloração de Gram, testes de oxidase, catalase e coagulase. Posteriormente, foram analisados os espectros de proteínas gerados no MALDI-TOF MS, seguida pela separação e detecção de íons pelo tempo de voo (TOF).Foram identificadas sete espécies, sendo a de maior ocorrência S. chromogenes 65 (76%), seguida por, S. hyicus 5 (6%), S. epidermidis 4 (5%).A técnica de MALDI-TOF demonstrou-se efetiva na caracterização de espécies de Staphylococcus causadores de mastite bovina.

Diagnóstico e propostas de ações visando a melhoria da qualidade microbiológica do leite cru recebido em um laticínio localizado no Sul de Minas Gerais / Diagnosis and proposed actions aiming the improvement of the microbiological quality of raw milk received in a dairy located in the South of Minas Gerais

O objetivo deste estudo foi verificar a qualidade microbiológica do leite cru refrigerado recebido no Laticínio de uma Cooperativa localizada no Sul do Estado de Minas Gerais e propor ações corretivas. Amostras de leite foram coletadas dos compartimentos dos tanques de caminhões em três dias diferentes, plaqueadas em duplicata em Ágar Padrão para Contagem e incubadas a 7ºC por 10 dias. Foram identificadas 92 espécies através de espectrofotometria de massas MALDI-TOF-MS. Entre os isolados identificados o gênero Pseudomonas foi predominante, sendo conhecido por possuir grande capacidade de produção de enzimas degradativas em leite cru refrigerado. Foi possível verificar que somente o resfriamento do leite não garante a sua qualidade microbiológica, sendo necessárias outras ações visando à redução da contaminação microbiológica inicial do leite cru.

Sugarcane bagasse hydrolysis using yeast cellulolytic enzymes.

Ethanol fuel production from lignocellulosic biomass is emerging as one of the most important technologies for sustainable development. To use this biomass, it is necessary to circumvent the physical and chemical barriers presented by the cohesive combination of the main biomass components, which hinders the hydrolysis of cellulose and hemicellulose into fermentable sugars. This study evaluated the hydrolytic capacity of enzymes produced by yeasts, isolated from the soils of the Brazilian Cerrado biome (savannah) and the Amazon region, on sugarcane bagasse pre-treated with H2SO4. Among the 103 and 214 yeast isolates from the Minas Gerais Cerrado and the Amazon regions, 18 (17.47%) and 11 (5.14%) isolates, respectively, were cellulase-producing. Cryptococcus laurentii was prevalent and produced significant ß- glucosidase levels, which were higher than the endo- and exoglucanase activities. In natura sugarcane bagasse was pre-treated with 2% H2SO4 for 30 min at 150oC. Subsequently, the obtained fibrous residue was subjected to hydrolysis using the Cryptococcus laurentii yeast enzyme extract for 72 h. This enzyme extract promoted the conversion of approximately 32% of the cellulose, of which 2.4% was glucose, after the enzymatic hydrolysis reaction, suggesting that C. laurentii is a good ß-glucosidase producer. The results presented in this study highlight the importance of isolating microbial strains that produce enzymes of biotechnological interest, given their extensive application in biofuel production.

Avaliação da qualidade higiênico-sanitária dos sucos de laranja comercializados em Alfenas-MG / Evaluation of the hygienic quality of orange juice commercialized in Alfenas-MG

O suco de laranja é um alimento rico em nutrientes e muito consumido em diferentes partes do mundo. A pesquisa de bactérias indicadoras de contaminação auxilia na verificação da qualidade higiênico-sanitária do alimento. As causas mais freqüentes de contaminação dos sucos de laranja são: o controle inadequado da temperatura de estocagem; higiene insatisfatória do ambiente e de maquinários; contaminação cruzada e despreparo do manipulador. Desta forma, o monitoramento microbiológico do produto torna-se muito importante para verificar sua inocuidade. O presente trabalho avaliou a qualidade higiênico-sanitária de sucos de laranja comercializados de Alfenas-MG, comparando os sucos mantidos em refresqueiras com os sucos preparados na hora.(...) De acordo com a RDC n.12/2001, observou-se que 52% das amostras apresentaram NMP de coliformes a 35ºC acima do permitido e 14% das amostras apresentaram NMP de coliformes a 45ºC acima do limite estabelecido pela referida legislação. Observou-se também que os sucos naturais de refresqueiras apresentaram uma maior quantidade de coliformes 35ºC e a 45ºC quando comparados com os sucos naturais preparados na hora.