Efeito do probiótico lactobacilus casei adjunto a terapia periodontal em modelo experimental de periodontite: uma relação entre periodonto e intestino / Effect of the probiotic lactobacilus casei as an adjunct to periodontal therapy in an experimental model of periodontitis: a relationship between periodontium and intestine

A periodontite é uma condição crônica inflamatória que pode influenciar a microbiota intestinal. O tratamento padrão ouro para a periodontite inclui a raspagem e o alisamento corono-radicular (RACR), porém em casos complexos pode-se utilizar terapias adjuvantes, como os probióticos. A utilização deste tratamento adjuvante poderá contribuir para a melhoria da condição periodontal e a simbiose intestinal. O objetivo deste estudo foi avaliar o efeito na inflamação periodontal e intestinal da utilização do Lactobacillus casei (LC) adjunto a RACR em camundongos Balb/c com periodontite, induzida por ligadura. Este estudo é um ensaio pré-clínico, in vivo, randomizado, cego e controlado por placebo, constituído por 36 camundongos Balb/c machos. Os animais foram submetidos a indução da periodontite por colocação de ligadura com fio de seda 4.0 ao redor do segundo molar superior direito, sendo divididos em 4 grupos,o grupo controle: Sem Periodontite e sem RACR (n=8); Grupo Ligadura: Com Periodondite e sem RACR (n=10); Grupo raspagem: Com Periodontite e com RACR (n=10) ; Grupo raspagem + L.casei (n=8): Com Periodontite e com RACR + administração de LC, por gavagem, durante 30 dias. Foram realizadas análises de citocinas pelo método ELISA no tecido gengival (IL-6), intestinal (IL-1ß, IL-6 e IL-10) e sanguíneo (IL-1ß e IL-6), além de análises bioquímicas (TGO, TGP, ureia e creatinina) e contagem diferencial de leucócitos do sangue. Foram coletados fragmentos do intestino grosso desses animais e analisados quanto a biomarcadores do estresse oxidativo (SOD, GSH e MDA), atividade da acetilcolinesterase (AChE) e foi realizada contagem da população de Bactérias Produtoras de Ácido Láctico das fezes dos animais. A utilização do LC adjunto a RACR resultou em uma redução na expressão da IL-6 no tecido gengival de camundogos com Periodontite (p < 0,05). Para as inteleucinas séricas (IL-1ß e IL-6), não houve diferenças entre os grupos (p > 0,05). Já para as citocinas intestinais houve uma redução na expressão de IL-10 (p < 0,05), para os grupos em que foi induzida a Periodontite. Com relação ao estresse oxidativo intestinal os animais do Grupo raspagem e raspagem + LC tiveram uma redução dos níveis de MDA (p < 0,05), para a SOD e o GSH, não houve diferenças significativas entre os 4 grupos pesquisados (p < 0,05). Conclui-se que o uso de LC adjunto a RACR em camungongos com periodontite induzida por ligadura pode reduzir a liberação de IL-6 no tecido gengival. Com relação aos efeitos intestinais foi observada a modulação da resposta inflamatória, com a redução de MDA, nos animais que receberam o tratamento periodontal. E a redução da expressão da IL-10, nos animais com Periodontite (AU).

Periodontitis is a chronic inflammatory condition that can influence the gut microbiota. The gold standard treatment for periodontitis includes scaling and crown-root planing, but in complex cases adjuvant therapies such as probiotics can be used. The use of this adjuvant treatment may contribute to the improvement of periodontal condition and intestinal symbiosis. The aim of this study was to evaluate the effect on periodontal and intestinal inflammation of the use of Lactobacillus casei (LC) adjunct to scaling and root planing (RACR) in Balb/c mice with ligature-induced periodontitis. This study is a preclinical, in vivo, randomized, blinded, placebo-controlled trial consisting of 40 male Balb/c mice. The animals were submitted to periodontitis induction by placing a 4.0 silk suture ligature around the upper right second molar. The sample was divided into 4 groups, each with 10 animals: Group I: Without Periodontitis and without RACR; Group II: With Periodontitis and without RACR; Group III: With Periodontitis and with RACR; Group IV: With Periodontitis and with RACR + gavage of LC, for 30 days. Cytokine analyzes were performed by the ELISA method in gingival tissue (IL-6), intestinal tissue (IL-1ß, IL-6 and IL-10) and blood (IL-1ß and IL-6), the blood was also subjected to analysis biochemical (TGO, TGP, urea and creatinine) and differential leukocyte count. Fragments of the large intestine of these animals were collected and analyzed for biomarkers of oxidative stress (SOD, GSH and MDA), acetylcholinesterase (AChE) activity, and the population of Lactic Acid-Producing Bacteria in the animals' feces was counted. The use of LC adjunct to RACR resulted in a reduction in the expression of IL-6 in the gingival tissue of mice with Periodontitis (p < 0.05), for the blood inteleukins (IL-1ß and IL-6), there were no differences between the groups (p > 0.05). As for intestinal cytokines, there was a reduction in the expression of IL-10 (p < 0.05), for the groups that presented Periodontitis. Regarding intestinal oxidative stress, the animals in Groups III and IV had a reduction in MDA levels (p < 0.05), for SOD and GSH, there were no significant differences between the 4 groups studied (p < 0.05 ). It is concluded that the use of LC adjunct to RACR in mice with ligation-induced periodontitis can reduce the release of IL-6 in the gingival tissue. Regarding the intestinal effects, two effects were found: The first related to the modulation of the inflammatory response, with the reduction of MDA, in the animals that received periodontal treatment. And the second related to a pro-inflammatory effect, with the reduction of IL-10 expression (AU).

Selective Cytotoxic effect of Probiotic, Paraprobiotic and Postbiotics of L. casei strains against Colorectal Cancer Cells: Invitro studies

Abstract This study highlights the cytotoxic effect of three L. casei strains on colorectal cell lines in invitro conditions. Different concentrations of live, heat killed (HK) and cell free supernatant (CFS) of three L.casei strains were subjected to CaCo2 and MRC5 cell lines. The viability of the treated and untreated cells was determined after 72 hrs by MTT assay, and IC50 estimated. Apoptosis was evaluated by Annexin V-propidium iodide method using flow cytometry. The live, HK and CFS of the L. casei strains showed cytotoxic effects on colorectal cell lines with significant differences. The cytotoxicity effects of live cells on CaCo2 cells were significantly higher (p˂0.01) than the HK cells. A dose dependent response was observed, as higher concentrations resulted in enhanced cytotoxicity effects. Live L.casei 1296-2cells inhibited 91% of CaCo2 cell growth, with IC50 of less than 108 cfu/ml. MRS medium and concentrations of CFS at above 20% v/v, were cytotoxic to the normal cell lines. Flow cytometry analyses of L. casei 1296-2 indicated that cytotoxicity effects on CaCo2 cells is related to apoptotic induction. Invitro studies indicate that Live and CFS of L. casei 1296-2 might be promising candidate for the control of colorectal cancers

Aplicação do plasma de baixa temperatura sob pressão atmosférica no controle de biofilmes cariogênicos / Application of low temperature plasma under atmospheric pressure in the control cariogenic biofilms

O objetivo geral do presente estudo foi avaliar a aplicação dos jatos de plasma de baixa temperatura sob pressão atmosférica (PBTPA) produzidos por gás de argônio e hélio como gases de trabalho, no controle de biofilmes cariogênicos. Para tanto, foram estabelecidos os parâmetros físicos dos PBTPA gerados com argônio e hélio que se mostraram efetivos frente a biofilmes mono, dual e polimicrobianos compostos por combinações das espécies Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguinis, Lactobacillus casei, Lactobacillus acidophilus, Candida albicans e Actinomyces naeslundii. Os biofilmes mono, dual e multi-espécies foram submetidos ao tratamento com PBTPA produzidos por dois dispositivos diferentes, um obtido comercialmente (kINPen09®) que usou argônio como gás de trabalho, e outro protótipo desenvolvido pela FEG-UNESP (Faculdade de Engenharia de Guaratinguetá) que usou hélio. Análises quantitativas e microscópicas (confocal, microscopia eletrônica de varredura) foram realizadas. Foi incluído controle negativo (sem tratamento), positivo (clorexidina 0,12%) e controle de gás, utilizando apenas fluxo de gás, sem produzir plasma. Além disso, os efeitos celulares do PBTPAargônio e hélio sobre biofilme dual e multi-espécies também foram analisados em microscopia eletrônica de varredura e microscopia de varredura a laser confocal. Todos os ensaios foram realizados em triplicata em três experimentos independentes. Os resultados foram tabulados e analisados quanto à distribuição. A seguir, os testes estatísticos mais adequados foram selecionados. O nível de significância foi de 5%. Os resultados obtidos para os tratamentos dos biofilmes mono, dual ou multi-espécies com PBTPA-argônio e hélio foram todos significativos em comparação ao controle negativo em todos os tempos analisados. Para PBTPA-argônio, não houve recuperação de S. gordonii e S. sanguinis em todos tempos analisados. Para PBTPA-hélio, os melhores resultados foram obtidos em 5 e 7 minutos de exposição dos biofilmes ao PBTPA. Finalmente, tanto o dispositivo gerador de PBTPA que trabalhou com gás argônio quanto o dispositivo que trabalhou com gás hélio, demonstraram resultados promissores e poderão contribuir para o desenvolvimento de novos protocolos de Odontologia de Intervenção Mínima. (AU) The general objective of this study was to evaluate the application of lowtemperature plasma under atmospheric pressure (PBTPA) of argon and helium flow, in the control of cariogenic biofilms. For this, the effective physical parameters of PBTPA-argon and helium in mono, dual and polymicrobial biofilms composed of combinations of the species Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguinis, Lactobacillus casei, Lactobacillus acidophilus, Candida albicans and Actinomyces naeslundii were established. The multi-species biofilms were treated by different PBTPA generating devices, one obtained commercially (kINPen09®) that used argon as working gas, and another prototype developed by FEG-UNESP (Faculdade de Engenharia de Guaratinguetá) that used helium as working gas. Quantitative and microscopic analyzes (confocal, scanning electron microscopy) were performed. Negative control (no treatment), positive control (chlorhexidine 2%) and gas control (argon) were included. Besides that, cellular effects of PBTPA-argon and helium on dual and multi-species biofilms were analyzed by scanning electron microscopy (SEM) and confocal laser scanning microscopy. The results obtained for the treatments of mono, dual or multispecies biofilms with both PBTPA-argon and helium were all significant when compared to the negative control at all times analyzed. For PBTPA-argon, there was no recovery of S. gordonii and S. sanguinis at all analyzed times. For PBTPA-helium, the best results were obtained at 5 and 7 min of exposure of biofilms to PBTPA. All the tests were carried out in triplicate in three independent experiments. The results are tabulated and analyzed in terms of distribution. Next, the most suitable statistical tests were selected. The level of significance was 5%. The results obtained for the treatments of mono, dual or multi-species biofilms with PBTPA-argon and helium were all significant compared to the negative control at all analyzed times. Finally, both PBTPA generating could contribute to the development of new protocols for Minimal Intervention Dentistry (AU)

O objetivo geral do presente estudo foi avaliar a aplicação dos jatos de plasma de baixa temperatura sob pressão atmosférica (PBTPA) produzidos por gás de argônio e hélio como gases de trabalho, no controle de biofilmes cariogênicos. Para tanto, foram estabelecidos os parâmetros físicos dos PBTPA gerados com argônio e hélio que se mostraram efetivos frente a biofilmes mono, dual e polimicrobianos compostos por combinações das espécies Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguinis, Lactobacillus casei, Lactobacillus acidophilus, Candida albicans e Actinomyces naeslundii. Os biofilmes mono, dual e multi-espécies foram submetidos ao tratamento com PBTPA produzidos por dois dispositivos diferentes, um obtido comercialmente (kINPen09®) que usou argônio como gás de trabalho, e outro protótipo desenvolvido pela FEG-UNESP (Faculdade de Engenharia de Guaratinguetá) que usou hélio. Análises quantitativas e microscópicas (confocal, microscopia eletrônica de varredura) foram realizadas. Foi incluído controle negativo (sem tratamento), positivo (clorexidina 0,12%) e controle de gás, utilizando apenas fluxo de gás, sem produzir plasma. Além disso, os efeitos celulares do PBTPAargônio e hélio sobre biofilme dual e multi-espécies também foram analisados em microscopia eletrônica de varredura e microscopia de varredura a laser confocal. Todos os ensaios foram realizados em triplicata em três experimentos independentes. Os resultados foram tabulados e analisados quanto à distribuição. A seguir, os testes estatísticos mais adequados foram selecionados. O nível de significância foi de 5%. Os resultados obtidos para os tratamentos dos biofilmes mono, dual ou multi-espécies com PBTPA-argônio e hélio foram todos significativos em comparação ao controle negativo em todos os tempos analisados. Para PBTPA-argônio, não houve recuperação de S. gordonii e S. sanguinis em todos tempos analisados. Para PBTPA-hélio, os melhores resultados foram obtidos em 5 e 7 minutos de exposição dos biofilmes ao PBTPA. Finalmente, tanto o dispositivo gerador de PBTPA que trabalhou com gás argônio quanto o dispositivo que trabalhou com gás hélio, demonstraram resultados promissores e poderão contribuir para o desenvolvimento de novos protocolos de Odontologia de Intervenção Mínima. (AU)

Atividade imunomoduladora sinérgica de probióticos Bifidobacterium animalis e Lactobacillus casei em células Caco-2 infectadas com Escherichia coli enteroagregativa (EAEC) / Synergistic immunomodulatory activity of probiotics bifidobacterium animalis and lactobacillus casei in enteroaggregative escherichia coli (eaec)-infected caco-2 cells

ABSTRACT BACKGROUND: Enteroaggregative Escherichia coli (EAEC) is an E. coli pathotype that presents aggregative adhesion patterns on in vitro cultivated cells, mainly related to persistent diarrhea cases in children. EAEC virulence factors are important for host colonization and pathogeni­city. Intestinal epithelial cells (IECs) recognize pathogen-associated molecular patterns (PAMPs) and initiate an immune response. Several studies using in vivo and in vitro models emphasize the probiotic activity and immunomodulatory capacity of Lactobacillus and Bifidobacterium species. OBJECTIVE To evaluate the modulation of cytokine production by probiotics Bifidobacterium animalis and Lactobacillus casei in human intestinal Caco-2 cells exposed to different strains of EAEC. METHODS: Caco-2 cells were incubated with EAEC strains in the presence or absence of probiotics. The production of cytokines IL-8, TNF-α, IL-1β and IL-10 was evaluated in the supernatants by a sandwich enzyme-linked immunosorbent assay (ELISA). RESULTS: Cytokine production did not change when uninfected and EAEC-infected Caco-2 cells were exposed to probiotics separately. All EAEC induced a significant increase in IL-8 production by Caco-2 cells, but the probiotics, even together, could not reduce its production. On the other hand, the synergic activity of probiotic strains significantly increased TNF-α production but decreased the basal production of IL-1ß. Also, probiotics induced a significant increase in the production of the anti-inflammatory cytokine IL-10 during EAEC infection. CONCLUSION: Our results reinforce the synergistic immunomodulatory activity of probiotics during EAEC infection.

RESUMO CONTEXTO: Escherichia coli enteroagregativa (EAEC) é um patotipo de E. coli que apresenta o padrão de aderência agregativa em células cultivadas in vitro, sendo comumente relacionada a casos de diarreia persistente em crianças. Fatores de virulência presentes em EAEC são importantes para a colonização do hospedeiro e patogenicidade. As células epiteliais intestinais (IECs) reconhecem padrões moleculares associados a patógenos (PAMPs) e iniciam uma resposta imune. Vários estudos usando modelos in vivo e in vitro enfatizam a atividade probiótica e a capacidade imunomoduladora de espécies de Lactobacillus e Bifidobacterium. OBJETIVO: Este estudo avaliou a modulação da produção de citocinas pelos probióticos Bifidobacterium animalis and Lactobacillus casei em células intestinais humanas Caco-2 expostas a diferentes cepas de EAEC. MÉTODOS: As células Caco-2 foram incubadas com as cepas de EAEC na presença ou ausência dos probióticos. A produção das citocinas IL-8, TNF-α, IL-1β e IL-10 foi avaliada nos sobrenadantes por ELISA sanduíche. RESULTADOS: Não houve alteração na produção de citocinas quando as células não infectadas e as células infectadas com EAEC foram expostas aos probióticos separadamente. Todas as cepas de EAEC induziram aumento significativo na produção de IL-8 pelas células Caco-2, mas os probióticos, ainda que em conjunto, não foram capazes de reduzir a produção desta citocina. Por outro lado, as cepas de probióticos aumentaram significativamente a produção de TNF-α mas diminuíram a produção basal de IL-1ß. Além disso, os probióticos induziram um aumento significativo na produção da citocina anti-inflamatória IL-10 durante a infecção por EAEC. CONCLUSÃO: Nossos resultados reforçam a atividade imunomodulatória sinérgica dos probióticos durante a infecção de EAEC.

Espectroscopia de impedancia eléctrica, una herramienta para aplicaciones biotecnológicas con Lactobacillus casei ATCC 393 / Electrical impedance spectroscopy, a tool for biotechnological applications with Lactobacillus casei ATCC 393

RESUMEN La aplicación de espectroscopia de impedancia eléctrica (EIE), es una técnica que se utiliza para monitorear, detectar y cuantificar microorganismos de interés biotecnológico, con la medición de parámetros eléctricos de respuesta rápida de un medio inoculado a temperatura y agitación constante mediante electrodos sumergidos. Realizando una comparación del modelo de crecimiento y el recuento en placa con los parámetros eléctricos de respuesta, se puede dar una correlación para romper la barrera tecnológica entre la microbiología clásica y los métodos rápidos de detección. La comparación de ambas técnicas fue realizada para determinar el máximo crecimiento de Lactobacillus casei (L. casei) ATCC 393. Se encontró que tras la inoculación, después de 24 h en condiciones microaerofílicas (37 °C), el máximo crecimiento microbiano fue registrado por medio de la EIE, mediante los parámetros -Z- (29,1057) y Deg-Deg0 (24,555°). En contraste con la técnica de conteo en placa, el crecimiento máximo se estimó a las 9 h. Los datos experimentales obtenidos mediante la EIE fueron ajustados por un circuito RC en serie, posteriormente, las curvas generadas fueron ajustadas a los modelos de crecimiento de Gompertz y Boltzman. Usando la técnica de EIE, la impedancia del medio resultó el parámetro más eficiente para la estimación del pico máximo exponencial de crecimiento de L. casei. Se demostró que la EIE constituye una alternativa para la detección rápida de la concentración microbiana en procesos de producción de biomasa para la elaboración de productos alimenticios probióticos.

ABSTRACT The application of electrical impedance spectroscopy (EIS) is a technique used to monitor, detect and quantify microorganisms of biotechnological interest, with the measurement of electrical parameters of rapid response of a medium inoculated at temperature and constant agitation by submerged electrodes. By making a comparison of the growth model and the plate count with the electrical response parameters, a correlation can be made to break the technological barrier between classical microbiology and rapid detection methods. The comparison of both techniques was performed to determine the maximum growth of Lactobacillus casei (L. casei) ATCC 393. It was found that after inoculation, after 24 h under microaerophilic conditions (37 °C), the maximum microbial growth was recorded by medium of the EIE, using the parameters -Z- (29,1057) and Deg-Deg0 (24,555°). In contrast to the plate count technique, maximum growth was estimated at 9 h. The experimental data obtained through the EIE were adjusted by a series RC circuit; later, the generated curves were adjusted to the growth models of Gompertz and Boltzman. Using the EIE technique, the impedance of the medium was the most efficient parameter for the estimation of the maximum exponential growth peak of L. casei. It was demonstrated that the EIE constitutes an alternative for the rapid detection of the microbial concentration in biomass production processes for the elaboration of probiotic food products.

Symbiotic drink based on Brazil nuts (Bertholletia excelsa H.B.K): production, characterization, probiotic viability and sensory acceptance / Bebida simbiótica à base de castanha-do-Brasil (Bertholletia excelsa H.B.K): produção, caracterização, viabilidade probiótica e aceitação sensorial

ABSTRACT: Brazil nut is a typical oilseed from the Amazon region, of great economic and cultural importance. Due to its high nutritional value, it is interesting to be used as a matrix for new products' development. Thus, the present study aimed to develop and characterize a drink based on Brazil nuts fermented by Lactobacillus casei and added with inulin. After formulated, the drink was pasteurized, fermented, maturated, and stored under refrigeration for 28 days. Physical, chemical, microbiological, and sensory analyzes were performed before and after fermentation, at 7-day intervals. There was maintenance in the content of lipids and proteins and reduction in the concentrations of fibers, ash, total carbohydrates, and moisture. The fermented drink pH decreased significantly during storage, while the acidity and the soluble solids content increased from the 14th day. Regarding the color, there were significant changes in all parameters analyzed. In the fermented drink, the development of molds and yeasts, total and thermotolerant coliforms, and Salmonella were not verified, while the control drink deteriorated after 7 days. The probiotic L. casei had count ranging from 9.48 to 8.59 log CFU mL-1 during the 28 days. When exposed to in vitro gastrointestinal conditions, L. casei significantly reduced its concentration. However, it reached the enteric phase with counts greater than 6.00 log UFC mL-1. Sensorially, the drink reached intermediate scores (between 5.41 and 6.02). Therefore, the use of Brazil nuts proved to be viable for the development of a product free of animal-origin components, symbiotic, and of outstanding nutritional quality.

RESUMO: A castanha-do-Brasil é uma oleaginosa típica da região Amazônica, de grande importância econômica e cultural. Devido ao seu alto valor nutritivo, torna-se interessante a sua utilização como matriz para o desenvolvimento de novos produtos. Assim, o presente estudo teve como objetivo o desenvolvimento e caracterização de uma bebida à base de castanha-do-Brasil fermentada por Lactobacillus casei e adicionada de inulina. Após formulada, a bebida foi pasteurizada, fermentada, maturada e armazenada sob refrigeração por 28 dias. Foram realizadas análises físicas, químicas, microbiológicas e sensoriais antes e após a fermentação, em intervalos de sete dias. Observou-se uma manutenção no teor de lipídeos e proteínas e redução nas concentrações de fibras, cinzas, carboidratos totais e umidade. O pH da bebida fermentada reduziu significativamente ao longo do armazenamento, enquanto a acidez e o teor de sólidos solúveis aumentaram a partir do 14° dia. Em relação à cor, houve alterações significativas em todos os parâmetros analisados. Na bebida fermentada, não foi verificado o desenvolvimento de bolores e leveduras, coliformes totais e termotolerantes e Salmonella, enquanto a bebida controle se deteriorou após sete dias. O probiótico L. casei apresentou contagens variando de 9.48 a 8.59 log CFU mL-1 durante os 28 dias de armazenamento. Quando exposto às condições gastrointestinais in vitro, o L. casei diminui a sua concentração significativamente. Contudo, atingiu a fase entérica com contagens superiores a 6.00 log UFC mL-1. Sensorialmente, a bebida alcançou scores intermediários (entre 5.41 e 6.02). Dessa forma, a utilização da castanha-do-Brasil se mostrou viável para o desenvolvimento de um produto isento de componentes de origem animal, simbiótico e de destacável qualidade nutricional.

Development of stuffed coalho cheese in the traditional, lactose-free and probiotic-added formulations / Desenvolvimento de queijo de coalho trufado nas versões tradicional, delactosado e com adição de probiótico

ABSTRACT: The probiotic products from the dairy market have been extensively exploited as well as lactose-free derivatives, suitable for lactose intolerant individuals. This research aimed to manufacture stuffed cheese with Brazilian cream cheese and spices in three versions (traditional, lactose-free, and probiotic cheese) and evaluate their quality by physico-chemical and microbiological analyses, Lactic Acid Bacteria (LAB) viability, and acceptance and preference assays. Physico-chemical properties of the formulations were all in accordance with the standards for the fat content of the Ordinance No. 146 for the traditional (54.06%) and probiotic (45.45%) full-fat samples and free-lactose (39.28%) medium-fat sample. Regarding the microbiological safety, all the samples presented to be ready for consumption, and the LAB count were still viable after 42 days of storage in the probiotic formulation at the order of 108 log CFU/mL. The lactose-free formulation achieved the highest acceptance rates among the public regarding sensorial qualities with an average grade of 7.27 out of 9. In the end, the three types of stuffed coalho cheese were successful in total quality control and also the probiotic formulation with the desired viable count after storage and lactose-free formulation with the best acceptance rates by the public.

RESUMO: A crescente consciência dos consumidores sobre a relação existente entre saúde e dieta, tem despertado o interesse por alimentos que promovem efeitos benéficos ao ser humano. Desta maneira, o mercado de produtos lácteos com alegações probióticas passou a ser bastante explorado, assim como os delactosados, adequados ao público intolerante a lactose. O objetivo deste estudo foi produzir queijo de coalho trufado com requeijão e especiarias nas versões tradicional, delactosado e com adição de probiótico, sendo realizadas análises físico-química, microbiológicas, viabilidade das bactérias ácido láticas, teste de aceitação e preferência. As características físico-químicas dos queijos produzidos permaneceram dentro dos padrões da legislação brasileira. Os queijos desenvolvidos apresentaram segurança microbiológica para consumo e as bactérias ácido láticas revelaram-se viáveis no produto por um período de 42 dias. Considerado mais uma alternativa para o público com intolerância a lactose, o queijo coalho trufado delactosado demonstrou melhor aceitação sensorial em relação as demais versões.

Comparison of Lactic Acid Production by L. casei in Batch, Fed-batch and Continuous Cultivation, Testing the use of Feather Hydrolysate as a Complex Nitrogen Source

Abstract A comprehensive comparison of the main fermentation parameters, productivity, yield and final L-lactic acid concentration, obtained through batch, fed-batch and continuous cultivations using Lactobacillus casei CCDM 198 and a model cultivation medium was carried out. Using this data, a pulse-feed fed-batch process was established for testing chicken feather hydrolysate as a replacement for all complex nitrogen sources (yeast and beef extracts and peptone) in the medium. As comparably high values of productivity (about 4.0 g/L/h) and yield (about 98 %) were reached under all cultivation conditions, the maximum final L-lactic acid concentration (116.5 g/L), as achieved through pulse-feed fed-batch fermentation, was chosen as the most important criterion for process selection. Fed-batch cultivation with chicken feather hydrolysate as both a complex nitrogen source and a neutralizing agent for maintaining constant culture pH yielded half the concentration of L-lactic acid compared to the model medium. We demonstrate here that chicken feather hydrolysate has potential for use in the production of L-lactic acid but its utilization requires further optimization

Effects of co-culture supernatant of Lactobacillus casei and Bacillus subtiliis natto on intestinal microecology, mucosal barrier function and immune function in mice with antibiotic-associated diarrhea / 中国药科大学学报

@#The aim of this study was to investigate the effects of co-culture supernatant of Lactobacillus casei(LC)and Bacillus subtiliis natto(BN)on intestinal micro-ecology, mucosal barrier function and immune function in mice with antibiotic-associated diarrhea(AAD). The AAD mouse model was established and the normal saline, LC, BN and co-culture supernatants were administered, respectively, for 4 days. The general conditions of the mice during the intervention were observed. The thymus and spleen weight ratios of different intervention mice were compared. The histopathological changes of the proximal colon lesions were observed. The intestinal microecology, mucosal barrier function and immune function of each group were detected. The results showed that the mice in the model group showed poor mental state, decreased feeding intake and abnormal stool characteristics, which were aggravated with the prolongation of time. After intervention, the above-mentioned states of mice in each group were improved, with the best recovery for the co-culture group. Histopathological results showed that the intestinal wall of the model group was severely damaged and villus was shedding. Cellulose-like exudation, necrosis of epithelial cells and infiltration of inflammatory cells could be seen in the model group. The pathological changes mentioned above were improved after intervention, and the co-culture group had the best effect. Compared with the control group, the thymus and spleen weight ratio, microbial diversity(Shannon)index, richness(Chao)index, Lactobacillus number, Bifidobacterium number, secretory immunoglobulin IgA(sIgA)in intestinal mucosa, interleukin(IL)-2 and IL-2/IL-4, the relative expressions of tight junction related protein-1(ZO-1)and atresia protein(Occludin)in intestinal tissue of the model group were lower, while the number of enterobacteria, enterococcus number, serum diamine oxidase(DAO)bacterial ectopic rate and IL-4 in intestinal tissue were higher(P< 0. 05). Compared with the model group, the thymus, spleen weight ratio, Shannon, Chao index, Lactobacillus number, Bifidobacterium number, sIgA in intestinal mucosa, IL-2 and IL-2/IL-4, the relative expressions of ZO-1 and Occludin in intestinal tissue of the intervention groups were higher(P< 0. 05), and the co-culture group was higher than the LC group and the BN group(P< 0. 05). There was no significant difference between the LC group and the BN group(P> 0. 05). Compared with the model group, the number of enterobacteria, enterococcus, serum DAO, bacterial ectopic rate and intestinal IL-4 in each intervention group were lower(P< 0. 05), and the co-culture group was lower than LC group and BN group(P< 0. 05). There was no significant difference between LC group and BN group(P> 0. 05). There were no significant differences in serum DAO, bacterial ectopic rate, sIgA, IL-2, IL-4 levels and IL-2/IL-4 levels between the co-culture group and the control group(P> 0. 05). The results showed that LC and BN co-culture supernatant can effectively regulate intestinal micro-ecology of AAD mice, improve intestinal mucosal barrier function, and improve intestinal and global immune function.

Evaluation of lactase activity in probiotic cultures containing inulin rich plant extracts as prebiotic sources

@#Aims: The present study deals with the isolation and identification of lactase producing probiotic strains from camel and sheep milk, determination of the enzyme activity by β-galactosidase assay (Miller Assay) in the presence of garlic, peas, onion and leeks extracts containing inulin as a prebiotic component. Methodology and results: The two isolates were screened for lactase producing ability to degrade lactose on MRS agar at 37 °C. These were identified as Lactococcus lactis from camel (Marecha) milk and Lactobacillus casei from sheep (Kajli) milk through morphological and biochemical tests using MRS medium. The optimized pH and temperature of both strains were 6 and 35 °C, respectively. Among the three concentrations used (0.2%, 0.4%, 0.8%), the optimal concentration of inulin rich onion and leeks extracts was 0.8% for maximum growth of L. casei and of the peas extract for L. lactis growth. 0.2% garlic extract was more effective prebiotic source for L. lactis growth. 0.8% commercial inulin used as a positive control was less effective as compared to plant extracts used in the study. With o-nitrophenyl-β-Dgalactoside) used as a substrate in the enzyme assay, maximum lactase activity obtained with 0.8% concentration of garlic extract is 7.10 Miller Units as compared to the peas extract with 6.17 Miller Units from L. lactis. Lactobacillus casei has produced more lactase, 6.85 Miller units with onion extract than with leeks extract, 6.43 Miller Units. Pure commercial inulin used as a control has given maximum enzyme activity as 9.14 Miller Units at 0.2% concentration. Conclusion, significance and impact of the study: It is concluded that the extracted prebiotic may enhance lactase activity of the probiotics to supplement the development of food products for lactose intolerant patients.

Survival of microencapsulated Lactobacillus casei (prepared by vibration technology) in fruit juice during cold storage

Background: Foods including probiotics are considered "functional foods." As an alternative to dairy products, we investigated the behavior of Lactobacillus casei when exposed to low-pH fruit juice. Juices of fruits such as pineapple, raspberry, and orange were assessed. Free and microencapsulated forms of L. casei were compared, and the viability of the probiotic was evaluated under storage at 4°C for 28 d. Microbiological analyses were carried out to ensure a safe and healthy product for consumers who look for foods with probiotics from sources other than dairy. Results: Low pH affected L. casei survival during storage depending on the type of fruit juice. In the case of pineapple juice, some microcapsules were broken, but microcapsules recovered at the end of the storage period had 100% viability (2.3 × 107 CFU/g spheres). In the case of orange juice, more than 91% viability (5.5 × 106 CFU/g spheres) was found. In raspberry juice, viability decreased rapidly, disappearing at the end of the storage period, which was caused by the absorption of high concentrations of anthocyanin inside microcapsules more than low pH. Conclusion: Low pH affected the survival of L. casei under refrigeration; even when they were microencapsulated, acidic conditions impacted their viability. Although pH affects viability, its value is very sensitive and will depend on the type of fruit juice and its composition. Some fruit juices contain compounds used as substrates for Lactobacillus and other compounds with antimicrobial effects.

Differential Effects of Lactobacillus casei Strain Shirota on Patients With Constipation Regarding Stool Consistency in China

BACKGROUND/AIMS: Probiotics are expected to confer benefits on patients with constipation, but how probiotics act on constipated patients with variable stool consistencies remains unclear. We investigated the effect of Lactobacillus casei strain Shirota (LcS) on constipation-related symptoms, especially stool consistency, of constipated patients. METHODS: Constipated patients meeting the Rome III criteria were divided into 3 groups according to the Bristol Stool Form Scale (BSFS): hard (hard stool [HS], BSFS < 3), normal (normal stool [NS], ≤ 3 BSFS ≤ 4), and soft (soft stool [SS], 4 < BSFS ≤ 5) stools. Subjects in each group consumed a probiotic beverage containing 1010 colony-forming units of LcS daily for 28 days. RESULTS: LcS intervention significantly alleviated constipation-related symptoms and increased defecation frequency in all subjects. Four weeks of LcS supplementation softened the hard stools in HS, hardened the soft stools in SS, and did not alter the ideal stool consistency in NS. The short-chain fatty acid (SCFA) concentrations were highest in SS, followed by NS and HS. LcS intervention increased the stool SCFA levels in HS but reduced or did not alter the levels in NS and SS. LcS intervention increased the Pseudobutyrivibrio and Roseburia abundances in HS and decreased the Pseudobutyrivibrio abundance in SS. CONCLUSIONS: LcS supplementation improved the constipation-related symptoms in constipated subjects. Differences in baseline stool consistency could result in different anti-constipation effects of LcS intervention. LcS balanced the stool consistency—softened the HS and hardened the SS. These effects could be associated with modulation of the gut microbiota and SCFA production.

Efeitos do queijo probiótico com adição de Lactobacillus casei-01 na periodontite experimental em ratos / Effects of probiotic cheese supplemented with Lactobacillus casei-01 on experimental periodontitis in rats

Objetivo: Este estudo avaliou os efeitos da ingestão de alimento funcional, queijo Prato contendo Lactobacillus casei-01, no desenvolvimento da periodontite experimental (PE) em ratos. Materiais e Métodos: 44 ratos machos Wistar foram divididos em 4 grupos (n = 11): C (controle) - animais sem indução de PE e alimentados com ração convencional (RC) + queijo convencional (CONV); PROB - animais sem indução de PE e alimentados com RC + queijo probiótico (PROB); PE - animais com indução de PE e alimentados com RC + queijo CONV e PE-PROB - animais com indução de PE e alimentados com RC + queijo PROB. O queijo Prato foi fabricado por método tradicional com bactérias lácticas, suplementado ou não com a cepa probiótica Lactobacillus casei-01. No dia 0 do experimento, queijo CONV ou PROB, de acordo com o grupo experimental, foi administrado oralmente para todos os animais 1 x / dia até o final do experimento. No dia 28, a PE foi induzida nos primeiros molares inferiores direito e esquerdo dos animais dos grupos PE e PE-PROB. No dia 42, todos os animais foram submetidos à eutanásia. Foram realizadas análises microtomográfica na região da bifurcação e nos sítios vestibular, lingual e interproximais [volume ósseo (BV), número de trabéculas (Tb.N), espessura das trabéculas (Tb.Th), perda óssea alveolar (ABL)] e análises histológica (parâmetros dos tecidos moles e duros), histométrica [porcentagem de osso na furca (PBF)] e imunoistoquímica [fator de necrose tumoral-α (TNFα), interleucina (IL)-1ß, IL-10, fator de crescimento transformador-ß1 (TGFß1) e fosfatase ácida resistente ao tartarato (TRAP)] na região da furca. Foram realizadas análises histológica (parâmetros de inflamação, celularidade e estruturação tecidual) e histométrica [altura de vilosidades (VH) e profundidade de cripta (CD)] nos tecidos intestinais. Os dados foram submetidos à análise estatística. Resultados: Os valores de BV foram similares em todos os grupos experimentais. No entanto, os grupos PE e PE-PROB apresentaram Tb.N significativamente maior e Tb.Th significativamente menor que os grupos C e PROB. O grupo PE apresentou PBF significativamente menor que os grupos C e PROB, e similar ao grupo PE-PROB. Os grupos PE e PE-PROB apresentaram maior expressão de citocinas pró-inflamatórias (TNFα e IL-1ß) e TRAP que os grupos C e PROB. O grupo PE-PROB apresentou maior expressão de citocinas anti-inflamatórias (TGFß1 e IL-10) que o grupo PE. Não houve diferenças significativas para os valores de VH e CD entre os gru pos experimentais. Conclusão: O consumo de queijo Prato, tanto CONV como PROB, teve efeito protetor nos tecidos periodontais durante o desenvolvimento da PE em ratos. Contudo, o queijo PROB parece ter estimulado principalmente a expressão de citocinas antiinflamatórias, quando comparado ao queijo CONV, favorecendo o processo de reparo tecidual(AU)

Objective: This study evaluated the effects of a functional food (Prato cheese) intake supplemented with Lactobacillus casei-01 in experimental periodontitis (EP) in rats. Materials and Methods: 44 Wistar male rats were divided into 4 groups (n = 11): C (control) - animals without EP induction and fed with conventional feed (CF) + conventional cheese (CONV); PROB - animals without EP induction and fed with CF + probiotic cheese (PROB); EP - animals with EP induction and fed with CF + CONV cheese; and EP-PROB - animals with EP induction and fed with CF + PROB cheese. Prato cheese was produced by traditional lactic bacteria manufacturing method, supplemented or not with the probiotic strain Lactobacillus casei-01. On day 0, either CONV or PROB cheese, according to the experimental group, was orally administered to all animals once a day until the end of the experiment. On day 28, EP was induced on the right and left mandibular first molars of the animals of groups EP and EP-PROB. On day 42, all animals were euthanized. Microtomographic analysis was performed in the furcation region and at the buccal, lingual and interproximal sites [bone volume (BV), trabeculae number (Tb.N), trabeculae thickness (Tb.Th), alveolar bone loss (ABL)]. Histologic (hard and soft tissue parameters), histometric [bone percentage in the furcation area (PBF)] and immunohistochemical [transforming growth factor-α (TNF-α), interleukin (IL)-1ß, IL-10, transforming growth factor-ß1 (TGF- ß1) and tartrate resistant acid phosphatase (TRAP)] analyses were performed in the furcation region. Histologic (inflammatory, cellularity and tissue structure parameters) and histometric [villi height (VH) and crypt depth (CD)] analyses were performed in the intestinal tissues. Data were statistically analyzed. Results: BV values were similar in all experimental groups. However, groups EP and EP-PROB presented significantly greater Tb.N and significantly lower Tb.Th than groups C and PROB. Group EP presented significantly lower PBF than groups C and PROB, and similar to group EP-PROB. Groups EP and EP-PROB presented higher expression of pro-inflammatory cytokines (TNF-α and IL-1ß) and TRAP than groups C and PROB. Group EP-PROB presented higher expression of anti-inflammatory cytokines (TGFß1 e IL-10) than group EP. No significant differences were observed among groups for VH and CD values. Conclusion: CONV as well as PROB Prato cheese intake promoted a protective effect on periodontal tissues during EP induction in rats. However, PROB cheese seems to mainly stimulate the e xpression of antiinflammatory cytokines when compared with CONV cheese, favoring tissue repair process(AU)

Effect of Lactobacillus casei on the depressive behavior and the expression of NR1 and NR2A receptors in hippocampus of rats with postpartum depression induced by maternal separation / 中华行为医学与脑科学杂志

Objective@#To investigate the effect of Lactobacillus casei on depressive behavior and the expression of NR1, NR2A receptor in postpartum depression rats induced by maternal separation.@*Methods@#Twenty-four Sprague-Dawley female rats were randomly divided into control group(CON group) (n=8), postpartum depression group(PPD group) (n=8), postpartum depression with lactobacillus casei group(PPD+ Lactobacillus casei group) (n=8). Rats of the control group were given no interventions.Rats in PPD group and PPD+ Lactobacillus casei group were given maternal separation to establish depression model.And then the rats in PPD+ Lactobacillus casei group were treated with Lactobacillus casei(8×108CFU/(kg·d)) for 4 weeks after 14 days of maternal separation.The forced swimming test (FST) was employed to detect the depressive behaviors.Lactobacillus, Bifidobacterium, Enterococcus faecalis and Escherichia coli in cecum of rats and the expression of NR1 and NR2A mRNA in hippocampus of rats were detected by real-time fluorescence quantitative PCR.@*Results@#Compared with CON group ((157.50±8.13) s), the immobility time of PPD group((200.00±10.35) s) was significantly longer (t=-3.23, P<0.05). Compared with PPD group, the immobility time of PPD+ Lactobacillus casei group ((153.25±7.41) s) was significantly shortened (t=3.67, P<0.05), and the depressive behavior was improved.Compared with CON group, the mRNA expression of Lactobacillus ((1.47±0.08), t=-5.87, P<0.01), Enterococcus faecalis ((1.23±0.08), t=-2.85, P<0.05) and Escherichia coli( (1.33±0.07), t=-4.96, P<0.01) in caecum were significantly increased in PPD group, while that of Bifidobacterium decreased significantly((0.65±0.07), t=5.18, P<0.01). Compared with PPD group, the mRNA expression of Lactobacillus( (1.05±0.05), t=3.67, P<0.01), Enterococcus faecalis ((0.97±0.05), t=2.74, P<0.05) and Escherichia coli ((1.06±0.05), t=3.31, P<0.01) were significantly decreased in PPD+ Lactobacillus casei group, while that of Bifidobacterium increased significantly ((0.98±0.04), t=-4.26, P<0.01). Compared with CON group, the mRNA expression of NR1 receptor ((0.57±0.04), t=9.65, P<0.01) and NR2A receptor ((0.60±0.05), t=8.64, P<0.05) in hippocampus of rats in PPD group were significantly decreased.Compared with PPD group, the expression of NR1 receptor ((1.01±0.05), t=-5.39, P<0.01) and NR2A receptor ((0.98±0.07), t=-3.91, P<0.05) in hippocampus were increased significantly in PPD+ Lactobacillus casei group.@*Conclusion@#Lactobacillus casei can improve the depressive behavior of postpartum depression in rats, and improve the intestinal flora, which affect the expression of NR1 and NR2A in hippocampus.

Effect of Lactobacillus casei on the depressive behavior and the expression of NR1 and NR2A recep-tors in hippocampus of rats with postpartum depression induced by maternal separation / 中华行为医学与脑科学杂志

Objective To investigate the effect of Lactobacillus casei on depressive behavior and the expression of NR1, NR2A receptor in postpartum depression rats induced by maternal separation. Methods Twenty-four Sprague-Dawley female rats were randomly divided into control group( CON group) (n=8),postpartum depression group(PPD group) (n=8),postpartum depression with lactobacillus casei group(PPD+ Lactobacillus casei group) (n=8). Rats of the control group were given no interventions. Rats in PPD group and PPD+Lactobacillus casei group were given maternal separation to establish depression model. And then the rats in PPD+Lactobacillus casei group were treated with Lactobacillus casei ( 8× 108CFU/(kg·d)) for 4 weeks after 14 days of maternal separation. The forced swimming test ( FST) was employed to detect the depressive behaviors. Lactobacillus,Bifidobacterium,Enterococcus faecalis and Esche-richia coli in cecum of rats and the expression of NR1 and NR2A mRNA in hippocampus of rats were detec-ted by real-time fluorescence quantitative PCR. Results Compared with CON group ((157. 50±8. 13) s), the immobility time of PPD group((200. 00±10. 35) s) was significantly longer (t=-3. 23,P<0. 05). Com-pared with PPD group,the immobility time of PPD+ Lactobacillus casei group ((153. 25±7. 41) s) was sig-nificantly shortened (t=3. 67,P<0. 05),and the depressive behavior was improved. Compared with CON group,the mRNA expression of Lactobacillus ((1. 47± 0. 08),t=-5. 87,P<0. 01),Enterococcus faecalis ((1. 23±0. 08),t=-2. 85,P<0. 05) and Escherichia coli( (1. 33±0. 07),t=-4. 96,P<0. 01) in caecum were significantly increased in PPD group, while that of Bifidobacterium decreased significantly (( 0. 65 ± 0. 07),t=5. 18,P<0. 01). Compared with PPD group,the mRNA expression of Lactobacillus ( ( 1. 05 ± 0. 05),t=3. 67,P<0. 01),Enterococcus faecalis ((0. 97±0. 05),t=2. 74,P<0. 05) and Escherichia coli ((1. 06±0. 05),t=3. 31,P<0. 01) were significantly decreased in PPD+ Lactobacillus casei group,while that of Bifidobacterium increased significantly ((0. 98± 0. 04),t=-4. 26,P<0. 01). Compared with CON group,the mRNA expression of NR1 receptor (( 0. 57 ± 0. 04), t=9. 65, P<0. 01) and NR2A receptor ((0. 60±0. 05),t=8. 64,P<0. 05) in hippocampus of rats in PPD group were significantly decreased. Com-pared with PPD group,the expression of NR1 receptor ((1. 01±0. 05),t=-5. 39,P<0. 01) and NR2A re-ceptor ((0. 98±0. 07),t=-3. 91,P<0. 05) in hippocampus were increased significantly in PPD+ Lactoba-cillus casei group. Conclusion Lactobacillus casei can improve the depressive behavior of postpartum depression in rats,and improve the intestinal flora,which affect the expression of NR1 and NR2A in hippocampus.

Immunogenicity of the truncated NDV F protein surface-displayed on Lactobacillus casei / 生物工程学报

To evaluate immune efficacy of the recombinant Lactobacillus casei, we constructed pLA-Newcastle disease virus (NDV)-F/L. casei and obtained the expression products. PCR amplified the NDV F gene carrying part of the major epitopes. The target gene was inserted to the shuttle plasmid pLA, and then transformed into Escherichia coli BL21 (DE3) in order to screen positive recombinant plasmid. The positive recombinant plasmid was transformed into L. casei by electroporation to construct pLA-NDV-F/L. casei. The positive strains were identified by PCR. The reactivity of the recombinant bacteria was identified by Western blotting and the protein expression was detected by indirect immunofluorescence, flow cytometry and laser confocal microscopy. The 14-day-old chickens in each group were vaccinated by oral plus nose drops. The pLA-NDV-F/L. casei twice immunization group and three times immunization group, the commercial vaccine group, the pLA/L. casei group, the unchallenge PBS and the challenge PBS group were established. IgG in serum and sIgA in the lavage fluid of intestinal, nasal and lung were detected by ELISA. The protection rate of chickens was evaluated. The results showed that 94.10% of the recombinant bacteria expressed the F protein. The recombinant protein was highly expressed on the surface of L. casei with a protein size of 62 kDa, which specifically bound to anti-NDV serum. The levels of anti-F IgG and sIgA antibodies in each test group were significantly higher than those in the control groups. The duration of antibody in the pLA-NDV-F/L. casei three-time immunization group lasted 28 days longer than that in the twice immunized group, and there was no significant difference between antibody peak values. The attack protection rates in each group of immunized pLA-NDV-F/L. casei three times, twice, attenuated vaccine, pLA/L. casei and PBS were 80%, 80%, 90%, 0% and 0%, respectively. Therefore, the antigenic protein of NDV F was successfully expressed by L. casei expression system, which has of reactogenicity and immunogenicity, and could induce protective immune responses in chickens.

Risk Assessment of Lactobacillus casei W56 Used as "Other Substances"

The Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) has, at the request of the Norwegian Food Safety Authority (Mattilsynet; NFSA), assessed the risk of "other substances" in food supplements sold in Norway. These risk assessments will provide NFSA with the scientific basis while regulating the addition of “other substances” to food supplements and other foods. "Other substances" are described in the food supplement directive 2002/46/EC as substances other than vitamins or minerals that have a nutritional and/or physiological effect. It is added mainly to food supplements, but also to other foods. VKM has not in this series of risk assessments of "other substances" evaluated any claimed beneficial effects from these substances, only possible adverse effects. The present report is a risk assessment of Lactobacillus casei W56, and it is based on previous risk assessments and articles retrieved from a literature search. The risk of L. casei W56 was assessed for the general population. However, in previous assessments of “probiotics” published by VKM, concerns have been identified for specific groups. Therefore, the risk was assessed for the age group with immature gastro-intestinal microbiota (age group 0-36 months), population with mature gastro-intestinal microbiota (>3 years) and vulnerable groups independent of age. VKM has also assessed the risk of L. casei W56 in food supplements independent of the dose and have assessed exposure in general terms. Other sources of L. casei W56, such as foods, have not been included in the present risk assessment. VKM concludes that it is unlikely that L. casei W56 causes adverse health effects in the general healthy population with mature gastro-intestinal tract. However, no data on long-term adverse effects on infants and young children were identified. As evidence is accruing that the early microbial composition of the neonatal gut is important for the development of the gut microbiota and the immune system of the growing child, it is not possible to exclude that a daily supply of a single particular bacterial strain over a prolonged period of time to an immature gastro-intestinal tract may have long-term, although still unknown, adverse effects on that development.

Atividade antimicrobiana de Stevia rebaudiana Bertoni e de adoçantes não calóricos sobre bactérias cariogênicas: estudo in vitro / Antimicrobial activity of Stevia rebaudiana Bertoni and non-caloric sweeteners on cariogenic bacteria: in vitro study

Objetivo: avaliar a atividade antimicrobiana in vitro da planta Stevia rebaudiana Bertoni e de adoçantes não calóricos sobre o crescimento de Streptococcus mutanse Lactobacillus casei, micro-organismos cariogênicos presentes na cavidade bucal. Materiais e método: o estudo foi realizado utilizando as cepas padrões de S.mutans (UA159) e L. casei (ATCC7469). Foram avaliados diferentes compostos não calóricos substitutos dasacarose nas concentrações de 1%, 5% e 10%: eritritol(ER), Fit Sucralose® (SU), Stevita® (ST), solução de Steviarebaudiana Bertoni (SSr) e, como controle positivo,digluconato de clorexidina (DC). A análise do efeito inibitório desses compostos no crescimento das bactériasfoi feita por meio da técnica de difusão em ágar. Resultado:observou-se que existe um efeito inibitório decrescimento de ambos os micro-organismos por parte da SSr e do ER, enquanto os demais adoçantes testa dosnão tiveram efeito inibitório sobre esses micro-organismos.Conclusão: os resultados demonstram que SSR eER apresentam efeito inibidor no crescimento das cepastestadas de S. mutans e L. casei. (AU)

Objective: The study evaluated the in vitro antimicrobial activity of the Stevia rebaudiana Bertoni plant and non-caloric sweeteners on the growth of Streptococcus mutans and Lactobacillus casei, which are cariogenic microorganisms present in the oral cavity. Materials and method: The study was conducted using the standard strains of S. mutans (UA159) and L. casei (ATCC7469). Different non-caloric compounds were evaluated at concentrations of 1%, 5%, and 10%: erythritol (ER), Fit Sucralose™ (SU), Stevita™ (ST), Stevia rebaudiana Bertoni solution (SSr), and chlorhexidine digluconate (CD) as positive control. The inhibitory effect of these compounds on the growth of bacteria were analyzed by the agar diffusion technique. Result: There was a growth inhibition effect for both microorganisms by SSr and ER, whereas the other sweeteners tested had no inhibitory effect on the microorganisms. Conclusion: The results showed that SSr and ER present an inhibitory effect on the growth the strains tested of S. mutans and L. casei. (AU)

Effect of Arabic Gum as Prebiotics and Lactobacillus casei Shirota (LcS) as Probiotic on Oxidative Stress and Renal Function in Adenine–Induced Chronic Renal Failure in Rats

Objectives: This study aimed at assessing the effect of Gum Arabic as Prebiotics and lactobacillus casei shirota (LcS) as probiotic on oxidative stress and renal function in adenine–induced chronic renal failure in rats. Methodology: 70 male albino rats were divided into 7 groups and treated for 8 weeks as follows group 1: control basal diet group (BD), group 2: adenine in feed (0.75%, w/w), group 3: gum Arabic (GA) in drinking water (15%, w/v), group 4: lactobacillus casei shirota (LcS) 1 x 109 colony-forming units (CFU) supplement, group 5 adenine + GA, as before, group 6 adenine +(LcS) as before and group 7: adenine + GA+ (LcS) as before. Urine, blood and kidneys were collected from the rats at the end of the treatment for analysis of conventional renal function tests serum creatinine, urea, uric acid, sodium, potassium concentration). In addition, the oxidative stress markers serum and kidney glutathione and superoxide dismutase, serum catalase and malondialdhyde (MDA) were measured. Results: By the end of the 8 weeks of treatment, Adenine significantly (p <0.05) increased the concentrations of serum creatinine, urea, uric acid, sodium, potassium and serum MDA. In addition, the oxidative stress markers serum and kidney glutathione and superoxide dismutase, serum catalase was, significantly decreased. Treatment with (GA) and (LcS) significantly ameliorated these actions. The mechanism of the reported salutary effect of GA in adenine-induced CRF is associated with mitigation of the adenine- induced inflammation and generation of free radicals. Conclusion: The results suggest that that oral administration of gum Arabic and lactobacillus casei shirota could conceivably alleviate adverse effects of adenine induced renal toxicity (CRF).

Safety Evaluation of Lactobacillus casei C1 in Wistar Rats / Jurnal Sains Kesihatan Malaysia

Nowadays, probiotics have been widely consumed as supplementary food for their health benefits. However, safetyevaluation for many probiotic bacteria is still lacking. Furthermore, health benefits conferred by probiotics depend onthe strains used in producing probiotic products. Therefore, it is important to examine oral toxicity of newly isolatedLactobacillus casei (Lb. casei) C1. A total of 32 Wistar (WIS) rats were divided into acute (single dose) and subacuteoral toxicity (28-days repeated dose) groups. Rats in each group were further divided into control group which receivedphosphate buffer saline (PBS) orally and treatment group that was administered orally with Lb. casei C1 (1011 CFU/ml).For acute oral toxicity, treatment was performed on day-1 and the effects were monitored subsequently for 14 days. Forsubacute oral toxicity, treatment was given daily for 28 days and the effects were observed throughout the experimentalperiod. Body weight, food and water intake of the rats were recorded. Rats in acute and subscute groups were sacrificedon day-15 and day-29, respectively. Serum was collected to determine the levels of total protein, malondialdehyde (MDA),alanine transaminase (ALT), aspartate transaminase (AST), lactate dehydrogenase (LDH) and creatinine. Organs were alsoharvested for histological examination. There were no significant differences (p > 0.05) in body weight, food and waterintake between the control and treated rats in acute oral toxicity group. There were also no significant differences in theblood cell count, levels of total protein, MDA, LDH and creatinine between the control and treated rats. Similar findingswere recorded for the subacute oral toxicity group, except that the levels of ALT and AST which were significantly different(p < 0.05). When observed under a light microscope, there were no morphological changes detected in the kidney, liverand ileum of treated rats as compared to control rats in both of the experimental groups. In conclusion, Lb. casei C1exhibited no toxic effects in Wistar rats hence safe to be consumed orally.