Viabilidade celular de Saccharomyces cerevisiae cultivada em associação com bactérias contaminantes da fermentação alcoólica

O objetivo deste trabalho foi estudar a influência de bactérias dos gêneros Bacillus e Lactobacillus, bem como de seus produtos metabólicos, na redução da viabilidade celular de leveduras Saccharomyces cerevisiae. As bactérias Bacillus subtilis, Bacillus coagulans, Bacillus stearothermophilus, Lactobacillus fermentum e Lactobacillus plantarum foram cultivadas em associação com a levedura S. cerevisiae (cepa Y-904) por 72 horas a 32 °C, sob agitação. A viabilidade celular, a taxa de brotamento e a população de células de S. cerevisiae e a acidez total, a acidez volátil e o pH dos meios de cultivos foram determinados às 0, 24, 48 e 72 horas do cultivo misto. As culturas de bactérias foram tratadas através do calor, de agente antimicrobiano e de irradiação. Os resultados mostraram que apenas os meios de cultivo mais acidificados, contaminados com as bactérias ativas L. fermentum e B. subtilis, provocaram redução na viabilidade celular de S. cerevisiae. Excetuando a bactéria B. subtilis tratada com radiação gama, as demais bactérias tratadas pelos diferentes processos (calor, irradiação e antimicrobiano) não causaram diminuição da viabilidade celular e da população de S. cerevisiae, indicando que a presença isolada dos metabólitos celulares dessas bactérias não foi suficiente para reduzir a porcentagem de células vivas de S. cerevisiae.

Probiotics and oral health: In vitro and clinical studies

Aino Toiviainen Probiotics and oral health: in vitro and clinical studies University of Turku, Faculty of Medicine, Institute of Dentistry, Periodontology, Finnish Doctoral Program in Oral Sciences (FINDOS-Turku), Turku, Finland Annales Universitatis Turkuensis, Sarja – Ser. D, Medica-Odontologica. Painosalama Oy, Turku, Finland, 2015 Probiotics are used, for example, to prevent and treat diarrhea, allergies and respiratory infections, and there is an increasing interest to use probiotics also for oral health purposes. The most commonly used probiotic bacteria are lactobacilli and bifidobacteria, which are acidogenic and aciduric. From the oral point of view, use of these probiotics may, at least in theory, mean an increased risk of caries. In this thesis, the effects of probiotics on oral microbial composition, acid production of dental plaque and gingival health were studied through in vitro studies and two clinical studies. In a randomized, double-blind and crossover study, 13 healthy adults were allocated into two groups. Half of the subjects first consumed Lactobacillus rhamnosus GG tablets twice a day for two weeks, and after the washout period, L. reuteri tablets twice a day for two weeks. The other half of the subjects used the tablets in reverse order. In another controlled, randomized and double-blind study, 62 healthy adults were allocated into two groups. One group used the test tablets containing L. rhamnosus GG and B. lactis BB-12 and the other group used control tablets without probiotics. The recommendation for the use of the tablets was 4 per day for 4 weeks. Probiotic lactobacilli interfered with S. mutans biofilm formation and the adhesion of S. mutans to saliva-coated hydroxyapatite in vitro. No effect was found in S. mutans levels in the three-species biofilms. In clinical studies, the studied probiotics had no effect on the acid production of plaque. The counts of mutans streptococci and the oral microbial composition remained the same. Tablets containing L. rhamnosus GG and Bifidobacterium animalis subsp. lactis BB-12 did decrease the amount of plaque and gingival bleeding. According to our results, it seems that probiotics have beneficial effects on gingival health. The present results confirmed that probiotics are safe and have beneficial effects on oral health. Since the consumption of probiotics by the general population is steadily increasing, an understanding of the functions of probiotics in the oral cavity has become more important. Keywords: lactobacilli, bifidobacteria, caries, periodontal disease, mutans streptococci, probiotics

Evaluation of thermotolerant capacity of lactic acid bacteria isolated from commercial sausages and the effects of their addition on the quality of cooked sausages

The thermotolerant capacity of several lactic acid bacteria strains isolated from cooked commercial sausages was determined. Four strains were positively identified as Lactobacillus plantarum, Lactobacillus curvatus, Pediococcus pentosaceus and Pediococcus acidilacti, after surviving thermal treatment (70 °C during 60 minutes). Thermotolerant strains were inoculated in sausage batters before cooking in order to determine their effect on color, texture, acceptance and inhibition of Enterobacteria during 12 days at 8 °C. No significant effect of the inoculated strains was detected on color parameters. Textural profile parameters, cohesiveness and resilience, were not affected by the inoculation of thermotolerant lactic acid bacteria, but L. curvatus sausages resulted softer than the rest of the treatments. Samples inoculated with L. curvatus also obtained the lowest scores for the sensory attributes evaluated, with the remaining treatments causing no unfavorable effects on sausage acceptance. There was a reduction in enterobacterial counts after 12 days of cold storage in inoculated samples. The performance of inoculated lactic acid bacteria strains can be explained in a similar way as that of starter cultures in dry-fermented sausages, where the growth in nests impairs other pathogenic microorganisms present in the rest of the sausage, since environmental conditions and the early inoculation of these thermotolerant strains favor them to become the dominant flora.

Efeito da adição de probióticos sobre as características de queijo prato com reduzido teor de gordura fabricado com fibras e lactato de potássio

Leite pasteurizado com 1,3% de gordura foi utilizado na elaboração de queijo prato, no qual foi adicionado cultura probiótica de Lactobacillus rhamnosus e fibras, com substituição parcial do cloreto de sódio por solução de lactato de potássio a 60%. O objetivo do trabalho foi acompanhar o efeito da cultura probiótica adicionada no queijo prato fabricado com fibras e com substituição de cloreto de sódio por lactato de potássio, durante o período de 90 dias de armazenamento, a 6 °C. As análises físico-químicas e microbiológicas foram realizadas no queijo antes de ser embalado a vácuo e no 15, 30, 45, 60, 75 e 90 dia de armazenamento. O queijo com cultura probiótica, em todos os dias em que foi analisado, caracterizou-se por apresentar maior contagem de bactérias lácticas, valor de pH, teor de lactose e menor valor de acidez em relação ao queijo que não continha cultura probiótica.

Determinação da compatibilidade de desenvolvimento de culturas bacteriocinogênicas e fermento láctico

Além da utilização como bioconservantes de alimentos, algumas culturas bacteriocinogênicas estão sendo empregadas para acelerar a maturação de queijos. Porém a compatibilidade de desenvolvimento destas culturas com o fermento láctico é essencial para a obtenção de produtos característicos. O objetivo deste estudo foi avaliar a compatibilidade de desenvolvimento de Lactococcus lactis subsp. lactis ATCC 11454, Lactobacillus plantarum ALC 01 e Enterococcus faecium FAIR-E 198 com duas marcas comerciais de fermentos lácticos. Inicialmente, foi determinada a sensibilidade in vitro dos fermentos às culturas bacteriocinogênicas, somente Lc. lactis subsp. lactis ATCC 11454 foi capaz de promover a inibição de ambos os fermentos. Durante desenvolvimento associativo em leite a 35 ºC, as culturas bacteriocinogênicas não afetaram significativamente a produção de ácido láctico pelos fermentos. Estes, por sua vez proporcionaram aumento significativo da atividade de pediocina AcH e enterocina FAIR-E 198 e supressão da atividade da nisina. Dentre todas as culturas lácticas, Lb. plantarum ALC 01 apresentou a maior atividade de aminopeptidases (0,226 a 0,390). Portanto, baseado nos resultados em questão, Lb. plantarum ALC 01 e E. faecium FAIR-E 198 apresentam características de compatibilidade de desenvolvimento com o fermento mesofílico tipo O para serem empregadas como adjuntas no processamento de queijos.

Biocontrole de Listeria monocytogenes por Pediococcus acidilactici em couve minimamente processada

Este estudo avaliou um sistema de biocontrole para inibição de Listeria monocytogenes em couve minimamente processada, objetivando sua segurança durante estocagem sob refrigeração e em condições de abuso de temperatura. O potencial inibitório de bactérias láticas tolerantes ao sal e psicrotróficas contaminantes naturais da couve e Lactobacillus plantarum, Lactobacillus delbrueckii ATCC 9649 e Lactobacillus casei CCT 1465 foram avaliadas contra L. monocytogenes. O isolado de couve identificado como P. acidilactici CCA3 inibiu L. monocytogenes a 10 e 15 °C em ágar MRS e foi selecionado como possível agente de biocontrole. O número de L. monocytogenes na couve minimamente processada aumentou 3,7 e 4,7 ciclos logarítmicos a 5 e 10 °C, respectivamente, após 20 dias de armazenamento e 4,6 ciclos logarítmicos após oito dias a 15 °C. Entretanto, quando 10(8) UFC.g-1 de P. acidilactici CCA3 foram inoculados no produto processado, o crescimento de L. monocytogenes reduziu 2,3 ciclos logarítmicos sob temperatura abusiva de 15 °C. A acidez titulável e as características sensoriais da couve não foram alteradas pela presença de CCA3 ao longo do período de vida útil. Estes resultados sugerem o potencial de aplicação dos bioconservantes na couve minimamente processada, que necessitam estar associados à refrigeração e sanitização para garantir segurança.

Avaliação da metodologia microbiológica para determinação de 5-metiltetrahidrofolato em suco de caju (Anacardium occidentale L.)

Embora o suco de caju seja bastante consumido no Brasil e muitos estudos sobre seu valor nutricional tenham sido publicados, a literatura não reporta estudos sobre a determinação de folatos nesse suco. Várias técnicas analíticas podem ser utilizadas visando à detecção da referida vitamina. Optou-se por utilizar o ensaio microbiológico oficial de determinação de folatos em alimentos, utilizando-se Lactobacillus casei como microrganismo de teste. Foram testados a utilização de ultrassom e tratamento enzimático como fases preparatórias, a fim de determinar qual a melhor metodologia a ser empregada. Foram comparados também o uso de padrões de ácido fólico e folato. Os resultados mostraram que o emprego de enzima é indicado como fase preparatória na determinação desta vitamina em suco de caju. A utilização de ultrassom não interferiu, significativamente, nos resultados encontrados, sendo seu emprego dispensável. Os teores de folato encontrados no suco de caju integral in natura indicam que o consumo de uma porção de 200 mL do suco equivale à ingestão de aproximadamente 500 µg de folato, valor superior à ingestão diária recomendada para adultos.

Microbial profile of a kefir sample preparations: grains in natura and lyophilized and fermented suspension

Probiotics are supplementary foods developed by microbial strains that improve animal health beyond basic nutrition. Probiotics are consumed orally, regardless of being considered as normal inhabitants of the intestines, able to survive in enzimatic and biliary secretions. Kefir is a probiotic originated from the old continent, fermented by several bacteria and yeasts, encapsulated in a polyssacharide matrix, and resembles jelly grains. Kefir is also presented as its sourish product both in sugary or milky suspensions containing vitamins, aminoacids, peptides, carbohydrates, ethanol, and volatile compounds. Kefir is known to have a diverse microbial content depending on the country and fermentative substrates, which cause distinct probiotic effects. In this sense, the purpose of this work was to isolate, identify, and quantify the microbial content of a native sugary kefir sample (fermented suspension and lyophilized natural grains). Serial dilutions were plated on Rogosa agar (AR) and De Man, Rogosa and Sharpe (MRS), for Lactobacillus; Brain Heart Infusion (BHI), for total bacteria; Sabouraud-Dextrose-Agar (SDA), for yeasts and filamentous fungi; Thioglycolate Agar (TA), for Streptococcus, Acetobacteria and Leuconostoc; and Coconut Water Agar (CWA), and CWA supplemented with yeast extract (CWAY), for various genera. Genera and species for all strains were identified through biochemical reactions and specific API systems. The microbial profile of kefir was different from other sources of grains despite the presence of similar microorganisms and others which have not been reported yet. The data obtained with the CWA and CWAE media suggest that both substrates are alternative and salutary media for culture of kefir strains.

Caracterização taxonômica e fisiológica de bactérias ácido-láticas isoladas de pescado marinho

Bactérias ácido-láticas (BAL) constituem um grupo diverso, associado à produção de alimentos, incluindo fermentados de peixe. Por meio de processos fermentativos, é possível obter produtos mais saborosos e saudáveis. Estas bactérias também são importantes na medicina por suas atividades probióticas. Neste contexto, nove linhagens de bactérias ácido-láticas foram isoladas de sardinha e guaivira. Destas, duas linhagens foram caracterizadas fisiologicamente, juntamente com Leuconostoc mesenteroides ATCC8293. Lactobacillus homohiochii F3 apresentou ótimo crescimento a 40 ºC, diferente de Lactobacillus intestinalis G1 e Leuconostoc mesenteroides ATCC8293, que tiveram ótimo crescimento a 30 ºC. Em relação à tolerância ao NaCl, L. homohiochii F3 e L. mesenteroides ATCC8293 toleraram uma concentração de 5% de NaCl no meio. A espécie L. intestinalis G1 tolerou uma concentração de 7,5%, podendo ser considerada mais halotolerante que as demais espécies. L. mesenteroides ATCC8293, por causa do aumento da taxa de crescimento, pode ser considerada levemente halofílica. Na presença de concentrações não ótimas de NaCl, a adição de solutos orgânicos foi eficaz na restauração do balanço osmótico das células

Glycemic profile and prebiotic potential "in vitro" of bread with yacon (Smallanthus sonchifolius) flour

The purpose of this study was to elaborate bread with yacon flour at two different levels (6% and 11%) and to analyze their proximate composition, their glycemic indices and their prebiotic potentials in vitro. Bread with 6% and 11% of yacon flour presented, simultaneously, low and moderate glycemic index. As for the prebiotic potentials, it was evident the presence of probiotic bacteria, particularly Lactobacillus. The results showed that, the addition of yacon flour on bread rendered products from low to moderate GI, with prebiotic potential, low fat and high fiber contents, according to the Brazilian food legislation.

The influence of different combinations of probiotic bacteria and fermentation temperatures on the microbiological and physicochemical characteristics of fermented lactic beverages containing soybean hydrosoluble extract during refrigerated storage

Lactic beverages containing probiotics were prepared with whole UHT milk, whey of Mozzarella cheese, soybean hydrosoluble extract and sugar. Three formulations were studied, each one containing a different combination of probiotic/starter bacteria, fermented at two different temperatures (37 and 45 °C). The aim of this work was to verify the influence of these variables on the viability of probiotic microorganisms and on the physicochemical stability of lactic beverages during storage under refrigeration (21 days at 7 °C). The results indicated that the fermentation temperature had a significant effect on the viability of probiotic bacteria. Counts for Lactobacillus acidophilus were affected by storage time, resulting appropriate after 21 days only for the beverage fermented at 37 °C. Physicochemical parameters did not exhibit drastic variations - proving the stability of formulations during storage. Cells of Bifidobacterium spp. showed high survival ability, probably due to the presence of growth promoters from soybean and cheese whey. The fermentation temperature of 37 °C allowed counts above the minimum limit for all the studied microorganisms, being preferred to the temperature of 45 °C. The inclusion of soybean hydrosoluble extract, a prebiotics source, resulted in a symbiotic product with more benefits to the health of consumers.

Characterization of spoilage bacteria in pork sausage by PCR-DGGE analysis

To investigate microbial diversity and identify spoilage bacteria in fresh pork sausages during storage, twelve industrial pork sausages of different trademarks were stored at 4 ºC for 0, 14, 28 and 42 days, 80% relative humidity and packaged in sterile plastic bags. Microbiological analysis was performed. The pH and water activity (a w) were measured. The culture-independent method performed was the Polymerase Chain Reaction - Denaturing Gradient Gel Electrophoresis (PCR-DGGE). The culture-dependent method showed that the populations of mesophilic bacteria and Lactic Acid Bacteria (LAB) increased linearly over storage time. At the end of the storage time, the average population of microorganisms was detected, in general, at the level of 5 log cfu g-1. A significant (P < 0.005) increase was observed in pH and a w values at the end of the storage time. The PCR-DGGE allowed a rapid identification of dominant communities present in sausages. PCR-DGGE discriminated 15 species and seven genera of bacteria that frequently constitute the microbiota in sausage products. The most frequent spoilage bacteria identified in the sausages were Lactobacillus sakei and Brochothrix thermosphacta. The identification of dominant communities present in fresh pork sausages can help in the choice of the most effective preservation method for extending the product shelf-life.

Antimicrobial properties of lactic acid bacteria isolated from uruguayan artisan cheese

Uruguayan artisan cheese is elaborated with raw milk and non-commercial starters. The associated native microbiota may include lactic acid bacteria and also potentially pathogenic bacteria. Lactic acid bacteria were isolated from artisan cheese, raw milk, and non-commercial starter cultures, and their potential bacteriocin production was assessed. A culture collection of 509 isolates was obtained, and five isolates were bacteriocin-producers and were identified as Enterococcus durans,Lactobacillus casei, and Lactococcus lactis. No evidence of potential virulence factors were found in E. durans strains. These are promising results in terms of using these native strains for cheese manufacture and to obtain safe products.

Phisicochemical, sensory, and microbiological evaluation and development of symbiotic fermented drink

The goal of this study was to develop a symbiotic lacteous drink, evaluate its physicochemical and sensory characteristics, and verify the viability of Lactobacillus acidophilus in the drink. The milk serum-based drink consisted of 50% milk serum containing 10% saccharose, 25% powdered milk, 15% yacon pulp, and cultures of Lactobacillus acidophilus-La 5E and Bifidobacterium bifidum BB12. It was stored for up to 21 days under refrigeration. The milk serum-based drink was analyzed for protein, ether extract, total dietary fiber, total frutans, carbohydrate content, color, pH, acidity, and contamination by coliforms, and Salmonella sp. Coliforms and Salmonella sp were not detected, and L. acidophilus and B. bifidum provided satisfactory probiotic counts up to 21 days of storage under refrigeration. Lactobacillus acidophilus resistance to gastric acids and bile salts was detected only up to seven days of storage when evaluated in vitro. Sensory analysis and purchase viability were evaluated by consumers at 0, 7, 14, and 21 days of storage. Based on the analytical results and consumer evaluation, the drink was acceptable up to seven days of storage.

Production of probiotic fresh white cheese using co-culture with Streptococcus thermophilus

In this research, the probiotic Streptococcus thermophilus was inoculated into milk as co-culture to produce probiotic cheese. The effects of using Streptococcus thermophilus with other probiotic bacteria on cheese composition, and microbiological viability during 28 days of storage were investigated. Sensorial properties were determined only at 1st and 28th days of storage. The results showed that the use of Streptococcus thermophilus as co-culture in probiotic cheese production did not affect negatively the cheese components. Fat and dry matter properties of cheese weren't influenced by added probiotic bacteria. However, different level of pH, salt and lactic acid were detected. All probiotic bacteria were present in high levels throughout storage of cheeses, above 7 Log cfu.g- 1, threshold required for probiotic activity. Sensory panel showed that the highest average sensory evaluation points were recorded in cheeses made with Streptococcus thermophilus plus Lactobacillus casei, whereas other probiotic bacteria combinations had been affected less in regard to taste or appearance.