RESUMO
Alterations of the microbiota-gut-brain axis has been associated with intestinal and neuronal inflammation in Parkinson's disease (PD). The aim of this work was to study some mechanisms associated with the neuroprotective effect of a combination (MIX) of lactic acid bacteria (LAB) composed by Lactiplantibacillus plantarum CRL2130 (riboflavin overproducing strain), Streptococcus thermophilus CRL808 (folate producer strain), and CRL807 (immunomodulatory strain) in cell cultures and in a chronic model of parkinsonism induced with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in aged mice, and under levodopa-benserazide treatment. In vitro, N2a differentiated neurons were exposed to the neurotoxin 1-methyl-4-phenylpyridinium (MPP+) and treated with intracellular bacterial extracts or with conditioned media from BV-2 cells exposed to the bacterial extracts. In vivo, motor skills, tyrosine hydrolase (TH) in brain and cytokine concentrations in serum and in brain were evaluated. The study of the faecal microbiota and the histology of the small intestine was also performed. The results showed that the neuroprotective effect associated with LAB MIX administration did not interfere with levodopa-benserazide treatment. This effect could be associated with the antioxidant and immunomodulatory potential of the LAB selected in the MIX, and was associated with the significant improvement in the motor tests and a higher number of TH + cells in the brain. In addition, LAB MIX administration was associated with modulation of the immune response. LAB administration decreased intestinal damage with an increase in the villus length /crypt depth ratio. Finally, the administration of the LAB MIX in combination with levodopa-benserazide treatment was able to partially revert the intestinal dysbiosis observed in the model, showing greater similarity to the profiles of healthy controls, and highlighting the increase in the Lactobacillaceae family. Different mechanisms of action would be related to the protective effect of the selected LAB combination which has the potential to be evaluated as an adjuvant for conventional PD therapies.
Assuntos
Benserazida , Levodopa , Camundongos Endogâmicos C57BL , Fármacos Neuroprotetores , Transtornos Parkinsonianos , Animais , Levodopa/farmacologia , Benserazida/farmacologia , Benserazida/uso terapêutico , Fármacos Neuroprotetores/uso terapêutico , Fármacos Neuroprotetores/farmacologia , Transtornos Parkinsonianos/tratamento farmacológico , Transtornos Parkinsonianos/metabolismo , Masculino , Camundongos , Combinação de Medicamentos , Microbioma Gastrointestinal/efeitos dos fármacos , Modelos Animais de Doenças , Lactobacillales , Probióticos/uso terapêutico , Antiparkinsonianos/farmacologia , Antiparkinsonianos/uso terapêutico , Streptococcus thermophilus/efeitos dos fármacosRESUMO
In recent years the relationship between the intestinal microbiota, the host and chronic non-communicable diseases has brought interest into the study of its formation and maintenance in the host. Lactic acid bacteria (BAL) are Gram-positive bacteria with probiotic activity, which have been associated with many health benefits, such as decreased body fat mass and lower risk of type II diabetes mellitus. One of the main colonization mechanisms and bacteria survival strategies is the production of biofilms and the use of prebiotics as substrates to achieve a balance within intestinal microbiota. However, there is not enough evidence to demonstrate the biofilm formation in the presence of agave fructans (AF). This study aimed to evaluate in vitro the biofilm formation in a consortium of lactic acid bacteria: Lactobacillus delbrueckii ssp. lactis, Lactobacillus delbrueckii ssp. bulgaricus y Streptococcus thermophilus in the presence of AF at different concentrations: 0%, 0,1%, 4%, 8% y 16%. The addition of 0,1% of AF correlates with the best capacity for biofilm formation. The findings imply the possibility of modulating the biofilm formation of lactic acid bacteria with AF. These results can contribute positively to the host, by generating intestinal homeostasis, colonization resistance, stability to food digestion and chemical modifications of drugs and carry out beneficial functions to the health.
Assuntos
Agave , Biofilmes , Frutanos , Lactobacillus delbrueckii , Probióticos , Streptococcus thermophilus , Biofilmes/crescimento & desenvolvimento , Biofilmes/efeitos dos fármacos , Lactobacillus delbrueckii/fisiologia , Streptococcus thermophilus/fisiologia , Frutanos/farmacologia , Agave/microbiologia , Consórcios Microbianos/fisiologiaRESUMO
The growth of the lactic acid bacteria (LAB), Streptococcus thermophilus and Lactobacillus bulgaricus, widely used for yogurt production, results in acid production and the reduction of the milk [Formula: see text]. Industrial processes can show temperature ([Formula: see text]) changes due to the large scale of the equipment. As [Formula: see text] and [Formula: see text] affect the LAB growth, this study aimed to model the dependence of S. thermophilus and L. bulgaricus as a function of temperature and pH and to estimate and internally validate their growth parameters and confidence intervals with different modeling approaches. Twenty-four datasets regarding the growth kinetics of S. thermophilus and L. bulgaricus were used for estimating the kinetic parameters for each pure culture. The classical Baranyi and Roberts (sigmoidal) primary and Rosso and coworkers (cardinal parameter) secondary models successfully described the experimental data. The one-step modeling approach showed better statistical results than the two-step approach. The values of eight growth parameters ([Formula: see text], [Formula: see text], [Formula: see text], [Formula: see text], [Formula: see text], [Formula: see text], [Formula: see text], and [Formula: see text]) for each culture estimated from the fitting with the one-step approach and the Monte-Carlo-based approach were similar. Low averaged root-mean-squared errors ([Formula: see text]) (0.125 and 0.090 log CFU/mL) and percent discrepancy factor [Formula: see text] ([Formula: see text] and [Formula: see text]) values for S. thermophilus and L. bulgaricus were obtained in the internal model validation, reinforcing the predictive ability of the model.
Assuntos
Lactobacillus delbrueckii , Streptococcus thermophilus , Lactobacillus , Temperatura , Concentração de Íons de Hidrogênio , FermentaçãoRESUMO
Despite functional goat milk products having emerged due to their importance for human nutrition and health, few studies have assessed the safety of consumption of goat dairy products containing potentially probiotic autochthonous lactic acid bacteria supplemented with prebiotic carbohydrates. Aiming this field, this study evaluated the safety of goat's milk fermented with Streptococcus thermophilus QGE, the autochthonous Limosilactobacillus mucosae CNPC007 culture, and the prebiotic inulin, through single- and repeated-dose oral toxicity tests (SDT and RDT, respectively) in animals. Ten female Swiss Webster mice were used for SDT evaluation - 2 groups, SDTc (20 mL/kg of filtered water) and SDTt (20 mL/kg of fermented milk) - and 40 Wistar rats for RDT - RDT3, RDT6, and RDT12 (treated with fermented milk at doses of 3 mL/kg, 6 mL/kg, and 12 mL/kg, respectively) and also RDTc (12 mL/kg of filtered water). For SDT, no signs of mortality or toxicity were observed, and the animals maintained the expected weight gain and feed intake. The RDT trials did not show mortality or signs of toxicity, as well as no change in body weight and organs, in the hematological and biochemical parameters, and also in relation to morphology and histology. Since the fermented milk did not cause any toxic effect in the conditions evaluated, it can be said that its no-adverse effect level (NOAEL) was considered to be higher than 20 mL/kg/day. Thus, the fermented milk with L. mucosae CNPC007 and inulin was considered to be of low toxicity, safe for use in rodents, and allowed for use in further studies.
Assuntos
Produtos Fermentados do Leite , Probióticos , Animais , Humanos , Ratos , Camundongos , Feminino , Leite/microbiologia , Prebióticos , Inulina/metabolismo , Streptococcus thermophilus/metabolismo , Técnicas de Cocultura , Fermentação , Ratos Wistar , Cabras , Água , Produtos Fermentados do Leite/microbiologiaRESUMO
BACKGROUND: Four commercial starter cultures containing Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus (S1, S2, S3 and S4; S3 also contained Limosilactobacillus fermentum) were compared for fermentation, volatile flavor compounds, physicochemical parameters and microbiology, in yogurt prepared from three milk base formulations with increased protein (B1, B2 and B3). RESULTS: The fermentation patterns differed among starters, with Yoflex Mild 1.0 (S4) and SLB95 (S2) showing the longest fermentation time, depending on the formulation. At 21 days, S. thermophilus counts were similar among starters and higher than 8.52 log CFU mL-1 , for all yogurts. The highest counts (6.86 log CFU mL-1 ) for L. delbrueckii subsp. bulgaricus was found for S2 yogurts made from whey protein hydrolysate (B3). Minor water-holding capacity was detected for YF-L811 (S1) yogurts. Yoflex Harmony 1.0 (S3) starter containing Lim. fermentum produced a distinctive volatile profile characterized by aldehydes with respect to yogurts prepared with S1, S2 and S4, which were characterized by ketones. CONCLUSION: Results indicate the usefulness of carrying out studies similar to the present one to select the most appropriate process conditions depending on the desired product. © 2022 Society of Chemical Industry.
Assuntos
Lactobacillus delbrueckii , Limosilactobacillus fermentum , Iogurte , Fermentação , Streptococcus thermophilusRESUMO
Brazilian artisanal cheeses date from the first Portuguese settlers and evolved via local factors, resulting in unique products that are now part of the patrimony and identity of different Brazilian regions. In this study, we combined several culture-independent approaches, including 16S/ITS metagenetics, assembly- and deep profiling-metagenomics to characterize the originality of the microbiota of five varieties of Brazilian artisanal cheeses from the South and Southeast regions of Brazil. Their core microbiota contained mainly lactic acid bacteria (LAB), of which Lactococcus lactis subsp. lactis was the most frequent, followed by Streptococcus thermophilus in the South region. Moreover, several samples from the Southeast region contained, as dominant LAB, two other food Streptococci belonging to a new species of the salivarius group and S. infantarius. Rinds of samples from the Southeast region were dominated by the halotolerant bacterium Corynebacterium variabile, and the yeasts Diutina catenulata, followed by Debaryomyces hansenii and Kodamaea ohmeri. Rinds from the South region contained mainly LAB due to their short ripening time, and the predominant yeast was D. hansenii. Phylogenomic analysis based on L. lactis metagenome-assembled genomes (MAGs) showed that most Brazilian strains are closely related and form a different clade from those whose genomes are available at this time, indicating that they belong to a specific group. Lastly, functional analysis showed that S. infantarius acquired a â¼ 26 kb DNA fragment from S. thermophilus starter strains that carry the LacSZ system, allowing fast lactose assimilation, an adaptation advantage for growth in milk. Finally, our study identified several areas of concern, such as the presence of somatic cell DNA and high levels of antibiotic resistance genes in several cheese microbiota, suggesting that milk from diseased animals may still be used occasionally. Overall, the data from this study highlight the potential value of the traditional and artisanal cheese production network in Brazil, and provide a metagenomic-based scheme to help manage this resource safely.
Assuntos
Queijo , Lactobacillales , Lactococcus lactis , Animais , Biodiversidade , Brasil , Queijo/análise , Microbiologia de Alimentos , Lactobacillales/genética , Lactococcus lactis/genética , Metagenômica , Streptococcus thermophilus/genética , LevedurasRESUMO
Fifteen samples of whey protein concentrate (WPC) were tested against 37 commercial Streptococcus thermophilus strains to detect infective bacteriophages. Seventy-three diverse phages were isolated from 12 samples, characterized by using DNA restriction patterns and host range analyses. Sixty-two of them were classified as cos, two as pac, and nine as 5093, according to PCR multiplex assays. Phage concentration was greater than 104 PFU/g for 25.3% of isolated phages. Seven phages showed an unusual wide host range, being able to infect a high number of the tested strains. Regarding thermal resistance, pac phages were the most sensitive, followed by cos phages, those classified as 5093 being the most resistant. Treatments at 85 °C for 5 min in TMG buffer were necessary to completely inactivate all phages. Results demonstrated that the use, without control, of these whey derivatives as additives in dairy fermentations could be a threat because of the potential phage infection of starter strains. In this sense, these phages constitute a pool of new isolates used to improve the phage resistance of starter cultures applied today in the fermentative industry.
Assuntos
Bacteriófagos , Fagos de Streptococcus , Bacteriófagos/genética , Indústria de Laticínios , Streptococcus thermophilus/metabolismo , Soro do Leite , Proteínas do Soro do Leite/metabolismoRESUMO
O objetivo deste estudo foi desenvolver uma formulação de bebida láctea bubalina probiótica adicionada de polpa de morango, comparando os efeitos do uso do leite de búfala e de vaca na elaboração dos produtos e verificando a possibilidade de suplementação com triptofano nos produtos lácteos probióticos. Como primeira etapa do trabalho, bebidas lácteas probióticas foram elaboradas a partir de leite bubalino e bovino, fermentadas com Streptococcus thermophilus TA040, Lactobacillus bulgaricus LB340 e Lactobacillus acidophilus La5, e formuladas com 0, 25 e 50% de soro em sua formulação. As bebidas foram avaliadas quanto à cinética de fermentação das culturas láticas utilizadas, ao teor de proteína, gordura e sólidos totais não gordurosos, pós-acidificação, viabilidade das culturas fermentadoras e sua capacidade de sobrevivência ao estresse gastrointestinal in vitro. As bebidas lácteas bubalinas apresentaram resultados superiores as bebidas bovinas. O uso do leite de búfala na elaboração das bebidas lácteas promoveu benefícios quanto as culturas láticas presentes nos produtos, exercendo efeito protetivo e influindo na preservação da viabilidade das bactérias ao longo do armazenamento refrigerado e durante a simulação do estresse gastrointestinal in vitro. As bebidas lácteas elaboradas com 25% apresentaram os resultados mais próximos aos obtidos pelos produtos controle, sem adição de soro, sendo selecionadas para a segunda parte do estudo. Nesta etapa, as formulações de bebida láctea com 25% de soro, foram acrescidas de um preparado com polpa de morango e bebidas sem adição da fruta, utilizadas como controle. As bebidas lácteas bubalinas frutadas, apresentaram menor teor de gordura e melhores características reológicas, com maior viscosidade e consistência do que os produtos controle, sem afetar a pós-acidificação, o perfil de ácido graxo, assim como, a viabilidade e a resistência às condições de estresse gastrointestinal in vitro das culturas fermentadoras. A avaliação da possibilidade de suplementar lácteos probióticos com triptofano foi realizada em conjunto com a Universidade de Milão. Para isso, iogurtes probióticos receberam adição de triptofano antes ou após a fermentação, sendo avaliados com relação ao perfil de pós-acidificação, quantidade de triptofano nos produtos, número de células viáveis por plaqueamento e citometria de fluxo ao longo do armazenamento a 25° e 4°C. Complementarmente, a influência da presença do triptofano no crescimento e produção de compostos antimicrobianos pelas culturas láticas, também foi avaliada. A adição de triptofano após a fermentação dos iogurtes, que foram armazenados sob refrigeração (4°C), além de não afetar a pós-acidificação dos produtos, apresentou benefícios quanto a viabilidade L. acidophilus, redução do dano e aumento do número de células vivas, promovendo teor maior do aminoácido nos iogurtes. A presença do triptofano nos meios de cultivo, também influenciou de forma positiva o crescimento de S. thermophilus e L. acidophilus, melhorando o desenvolvimento das bactérias durante a fermentação e influindo em uma maior atividade antilistérica por parte do S. thermophilus. Diante da influência positiva da aplicação do leite de búfala na elaboração das bebidas lácteas, assim como, a adição do triptofano em iogurtes probióticos, a suplementação do aminoácido em bebidas lácteas bubalinas frutadas permitiria a obtenção de um produto funcional, onde seus benefícios estariam relacionados tanto ao consumo do probiótico presente no produto quanto a complementação de triptofano na dieta do consumidor
The aim of this study was to develop a formulation of probiotic buffalo dairy beverage added with strawberry pulp, comparing the effects of using buffalo and cow's milk in the preparation of products and verifying the possibility of tryptophan supplementation in probiotic dairy products. As a first stage of the work, probiotic dairy beverages were made from buffalo and bovine milk, fermented with Streptococcus thermophiles TA040, Lactobacillus bulgaricus LB340 and Lactobacillus acidophilus La5, and formulated with 0, 25 and 50% whey in their formulation. The beverages were evaluated for the fermentation kinetics of the used lactic cultures, the levels of protein, fat and total no fat solids, post-acidification, fermenting cultures viability and their ability to survive gastrointestinal stress in vitro. Buffalo milk use in dairy beverages production promoted benefits regarding the lactic cultures present in the products, exerting a protective effect and influencing the viability preservation of bacteria during the cold storage and simulation of gastrointestinal stress in vitro. Dairy beverages made with 25% whey addition showed results similar to those obtained by the control products, without whey addition, being selected for the second part of the study. In this part, the dairy beverages formulations with 25% whey, were added with a preparation were added with a strawberry pulp preparation and dairy beverages without added fruit, used as a control. Fruity bubaline dairy beverages had lower fat content and better rheological characteristics, with higher viscosity and consistency than control products, without affecting post-acidification, fatty acid profile, as well as viability and resistance to in vitro gastrointestinal condition of fermented cultures. The possibility of supplementing probiotic dairy products with tryptophan was evaluated in partnership with the University of Milan. For this, probiotic yogurts received the addition of tryptophan before or after fermentation, being evaluated in relation to the post-acidification profile, tryptophan amount in the products, viable cell number per plating and flow cytometry during storage at 25°C and 4°C. In addition, the influence of the tryptophan presence on the growth and production of antimicrobial compounds by lactic cultures was also evaluated. The addition of tryptophan after the yogurt fermentation, which were stored under refrigeration (4°C), in addition to not affecting the post-acidification of the products, showed benefits to the viability of L. acidophilus, reduced the damage and increased the number of cells promoting higher amino acid content in yogurts. Tryptophan presence in the culture media also positively influenced the growth of S. thermophiles and L. acidophilus, improving the development of bacteria during fermentation and influencing better antilisteric activity in the part of S. thermophiles. In view of the buffalo milk positive influence observed after the application in dairy beverage preparation, as well as the addition of tryptophan in probiotic yoghurts, amino acid supplementation in fruity buffalo dairy beverages would allow to obtain a functional product, where its benefits would be related both to the consumption of the probiotic present in the product as to the supplementation of tryptophan in the consumer's diet
Assuntos
Bebidas/efeitos adversos , Leite/efeitos adversos , Triptofano/classificação , Iogurte , Técnicas In Vitro/métodos , Búfalos , Contagem de Células/instrumentação , Química Farmacêutica , Probióticos/classificação , Streptococcus thermophilus/metabolismo , Lactobacillus delbrueckii/metabolismo , Crescimento e Desenvolvimento , Citometria de Fluxo/métodos , Soro do Leite/efeitos adversos , Frutas , Aminoácidos/antagonistas & inibidores , Lactobacillus acidophilus/metabolismoRESUMO
The profile of polyunsaturated fatty acids in cheeses obtained through fermentation by lactic acid bacteria Lactobacillus helveticus and Streptococcus thermophilus were evaluated. The milk used to make the cheeses came from cows fed with flaxseed oil and annato. The cheeses presented microbiological and physic-chemical quality with in the standards established by the legislation for Staphylococci and Listeria. With maturation, there was a reduction in the coliform values ââfor both treatments. Regarding the counts of lactic acid bacteria, these remained viable until the 30th day of maturation and the proteolytic bacteria decreased. For antioxidant capacity, the treatment containing the combination of the strains obtained high ABTS values. There was no significant difference between the treatments with respect to the color of the samples. For texture, there was a significant difference for the parameters cohesion and elasticity. No increase in CLA content was observed in the form of its two main isomers, however, the levels of polyunsaturated fatty acids were increased.
Assuntos
Queijo , Animais , Bovinos , Queijo/análise , Feminino , Fermentação , Microbiologia de Alimentos , Leite , Streptococcus thermophilusRESUMO
In fermented milks inoculated with two thermophilic strains (Lactobacillus bulgaricus and Streptococcus thermophilus), guabiroba pulp (Campomanesia xanthocarpa O. Berg) was added in different concentrations: 5% (I5 sample) and 10% (I10 sample), compared to a control sample, with no pulp addition. In these fermented milks, Bifidobacterium BB-12 was added and the samples were submitted to a progressive gastrointestinal simulation in vitro. The cells count was performed, including the survival rates for all the progressive steps of the simulated digestion. Total phenolic content (TPC) and antioxidant activity analysis by FRAP (Ferric Reducing Antioxidant Power) and DPPH (2,2-diphenyl-1-picrylhydrazyl) were performed in all the gastrointestinal steps. Before and during the entire gastrointestinal tract, the Bifidobacterium BB-12 count was 8-9 log CFU g-1, above the recommended for a probiotic product, with a highlight in intestinal colon steps. The I10 sample showed the highest viable cell count, the highest total phenolic content and antioxidant activity throughout the entire gastric steps (p < 0.05). The fermented milk proved to be an effective matrix for the probiotic stability and incorporation of guabiroba components. Bioactive compounds present in the guabiroba pulp may have occasioned a prebiotic and protective effect on Bifidobacterium BB-12 after gastric conditions. The possible bioconversion of these compounds in more active forms can contribute to the absorption in epithelial cells, enhancing fermented milks with guabiroba pulp as important sources of dietary accessible bioactive compounds.