Lactobacillus as probiotics: opportunities and challenges for potential benefits in female reproductive health.

The growing interest of the scientific community in the study of probiotics has gathered valuable data about its beneficial effects for multiple clinical conditions. This data also provides evidence for the functions and properties of probiotics and how they contribute to health benefits by influencing normal microbiota. Lactobacillus is an important genus which has long been utilized in the food industry and is also found as normal oral, intestinal and vaginal microbiota. Lactobacillus has shown multiple health benefits but its relative importance as a probiotic is majorly explored for gastrointestinal health. Healthy vaginal microbiota typically harbors Lactobacillus spp. providing several health benefits for female reproductive health, but there is more data required in order to compare the relative benefits with probiotic Lactobacillus added through either natural food sources or with standard probiotics supplements. The present article discusses the current status of knowledge about vaginal Lactobacillus as a probiotic and also compares the potential of probiotics from natural sources and through supplements along with recent approaches in this area.

Enterococcus faecium M7AN10 probiótico em matriz alimentar para cães / Enterococcus faecium M7AN10 probiotic in food matrix for dogs / Enterococcus faecium M7AN10 probiótico en matriz alimentaria para perros

Probióticos são capazes de melhorar o equilíbrio da microbiota intestinal, trazendo benefícios ao hospedeiro. Atualmente no mercado há poucas opções de alimentos, com probióticos em sua composição, destinados a cães e gatos. Portanto, o objetivo deste trabalho foi desenvolver uma matriz alimentar canina (ração úmida) com o probiótico Enterococcus faecium M7AN10. Para tal, avaliou-se a inocuidade, atividade enzimática, atividade antimicrobiana, potencial probiótico e a viabilidade do microrganismo em matriz alimentar canina. O isolado foi considerado inócuo, pois apresentou ausência de atividade hemolítica e de gelatinase, além de ser suscetível a diversos antimicrobianos. E. faecium M7AN10 apresentou atividade proteolítica e capacidade de produção de exoplissacarídeo. Em relação a atividade antimicrobiana pelo método da estria radial, o isolado inibiu Acinetobacter sp. 1, Corynebacterium sp. 4, Micrococcus luteus 33, Micrococcus luteus 43, Micrococcus sp. 3, Micrococcus sp. 20, Micrococcus sp. 36. Além disso, E. faecium M7AN10 apresentou capacidade de autoagregação de 33,50% e resistiu de forma constante quando submetido ao trato gastrointestinal in vitro em conjunto com Lacticaseibacillus rhamnosus LB 1.5 e Lacticaseibacillus paracasei LB 6.4. O cultivo misto manteve-se viável em matriz alimentar canina durante o período de oito dias. Com base nesses resultados, o isolado E. faecium M7AN10 foi considerada uma bactéria candidata a probiótico que pode vir a ser usada como aditivo em alimento para cães.

Probiotics are capable of improving the balance of the intestinal microbiota, bringing benefits to the host. Currently, on the market, there are few food options with probiotics in their composition intended for dogs and cats. Therefore, this research aimed to develop a canine food matrix (wet food) with the probiotic Enterococcus faecium M7AN10. To this end, the harmlessness, enzymatic activity, antimicrobial activity, probiotic potential, and viability of the microorganism in the canine food matrix were evaluated. The isolate was considered harmless, as it showed no hemolytic and gelatinase activity and was susceptible to several antimicrobials. E. faecium M7AN10 showed proteolytic activity and exopolysaccharide production capacity. Regarding antimicrobial activity using the radial stria method, the isolate inhibited Acinetobacter sp. 1, Corynebacterium sp. 4, Micrococcus luteus 33, Micrococcus luteus 43, Micrococcus sp. 3, Micrococcus sp. 20, Micrococcus sp. 36. Furthermore, E. faecium M7AN10 showed a self-aggregation capacity of 33.50% and resisted consistently when subjected to the gastrointestinal tract in vitro together with Lacticaseibacillus rhamnosus LB 1.5 and Lacticaseibacillus paracasei LB 6.4. The mixed culture remained viable in a canine food matrix over eight days. Based on these results, the isolate E. faecium M7AN10 was considered a candidate bacterium for a probiotic that could be used as an additive in dog food.

Los probióticos son capaces de mejorar el equilibrio de la microbiota intestinal, aportando beneficios al huésped. Actualmente en el mercado existen pocas opciones de alimentos con probióticos en su composición, destinados a perros y gatos. Por lo tanto, el objetivo de este trabajo fue desarrollar una matriz alimentaria canina (comida húmeda) con el probiótico Enterococcus faecium M7AN10. Para ello se evaluó la inocuidad, actividad enzimática, actividad antimicrobiana, potencial probiótico y viabilidad del microorganismo en matriz alimentaria canina. El aislado fue considerado inofensivo, ya que no mostró actividad hemolítica ni gelatinasa, además de ser susceptible a varios antimicrobianos. E. faecium M7AN10 mostró actividad proteolítica y capacidad de producción de exoplisacáridos. En cuanto a la actividad antimicrobiana mediante el método de las estrías radiales, el aislado inhibió a Acinetobacter sp. 1, Corynebacterium sp. 4, Micrococcus luteus 33, Micrococcus luteus 43, Micrococcus sp. 3, Micrococcus sp. 20, Micrococcus sp. 36. Además, E. faecium M7AN10 mostró una capacidad de autoagregación del 33,50% y resistió consistentemente cuando se sometió al tracto gastrointestinal in vitro junto con Lacticaseibacillus rhamnosus LB 1.5 y Lacticaseibacillus paracasei LB 6.4. El cultivo mixto permaneció viable en una matriz de alimento canino durante un período de ocho días. Con base en estos resultados, el aislado E. faecium M7AN10 se consideró una bacteria candidata para un probiótico que podría usarse como aditivo en la comida para perros.

Ultrasonic pre-treatment to enhance drying of potentially probiotic guava (Psidium guajava): Impact on drying kinetics, Lacticaseibacillus rhamnosus GG viability, and functional quality.

This study aimed to evaluate the effects of ultrasound (US) on the drying acceleration of potentially probiotic guava, including its impact on drying kinetics, probiotic (Lacticaseibacillus rhamnosus GG) viability, and functional quality of the product during drying. To perform US pre-treatments, one group of samples were first pre-treated by US (38 W/L, 25 kHz) for 15 and 30 min and then immersed in the probiotic solution for 15 or 30 min, and another group of samples were submerged in the probiotic solution simultaneously applying US (US-assisted) for 15 and 30 min. After pre-treatments, the samples were convectively dried at 60 °C. Based on the results, all US pre-treatments improved the drying rate (up to 59%) and reduced the drying time (up to 31%) to reach 25% moisture compared to non-sonicated samples. The reduction in drying time (from ∼6 h to ∼4 h for US pre-treated samples) was crucial for maintaining the probiotic viability in the dehydrated guavas. These samples showed counts of 6.15 to 7.00 CFU∙g-1 after 4 h, while the control samples reached counts of 4.17 to 4.45 CFU∙g-1 after 6 h. US pre-treatment did not affect the color parameters of the samples before drying (p > 0.05). The functional compounds were reduced during drying (p < 0.05), however, all US pre-treated samples had lower reductions in vitamin C content (up to 20%), phenolic compounds (up to 41%) and antioxidant capacity (up to 47%) compared to control samples (up to 52%, 81% and 61%, respectively). Therefore, US pre-treatment (highlighting the US-assisted probiotic incorporation for 30 min) reduced the drying time for guava slices and minimized the thermal impact on probiotic viability and functional compounds, being a strategy to produce potentially probiotic dehydrated guava.

Recent and Advanced DNA-Based Technologies for the Authentication of Probiotic, Protected Designation of Origin (PDO) and Protected Geographical Indication (PGI) Fermented Foods and Beverages.

The authenticity of probiotic products and fermented foods and beverages that have the status of protected designation of origin (PDO) or geographical indication (PGI) can be assessed via numerous methods. DNA-based technologies have emerged in recent decades as valuable tools to achieve food authentication, and advanced DNA-based methods and platforms are being developed. The present review focuses on the recent and advanced DNA-based techniques for the authentication of probiotic, PDO and PGI fermented foods and beverages. Moreover, the most promising DNA-based detection tools are presented. Strain- and species-specific DNA-based markers of microorganisms used as starter cultures or (probiotic) adjuncts for the production of probiotic and fermented food and beverages have been exploited for valuable authentication in several detection methods. Among the available technologies, propidium monoazide (PMA) real-time polymerase chain reaction (PCR)-based technologies allow for the on-time quantitative detection of viable microbes. DNA-based lab-on-a-chips are promising devices that can be used for the on-site and on-time quantitative detection of microorganisms. PCR-DGGE and metagenomics, even combined with the use of PMA, are valuable tools allowing for the fingerprinting of the microbial communities, which characterize PDO and PGI fermented foods and beverages, and they are necessary for authentication besides permitting the detection of extra or mislabeled species in probiotic products. These methods, in relation to the authentication of probiotic foods and beverages, need to be used in combination with PMA, culturomics or flow cytometry to allow for the enumeration of viable microorganisms.

Fermented Vegetables and Legumes vs. Lifestyle Diseases: Microbiota and More.

Silages may be preventive against lifestyle diseases, including obesity, diabetes mellitus, or metabolic syndrome. Fermented vegetables and legumes are characterized by pleiotropic health effects, such as probiotic or antioxidant potential. That is mainly due to the fermentation process. Despite the low viability of microorganisms in the gastrointestinal tract, their probiotic potential was confirmed. The modification of microbiota diversity caused by these food products has numerous implications. Most of them are connected to changes in the production of metabolites by bacteria, such as butyrate. Moreover, intake of fermented vegetables and legumes influences epigenetic changes, which lead to inhibition of lipogenesis and decreased appetite. Lifestyle diseases' feature is increased inflammation; thus, foods with high antioxidant potential are recommended. Silages are characterized by having a higher bioavailable antioxidants content than fresh samples. That is due to fermentative microorganisms that produce the enzyme ß-glucosidase, which releases these compounds from conjugated bonds with antinutrients. However, fermented vegetables and legumes are rich in salt or salt substitutes, such as potassium chloride. However, until today, silages intake has not been connected to the prevalence of hypertension or kidney failure.

Microencapsulation of Probiotics for Food Functionalization: An Update on Literature Reviews.

Functional foods comprise the largest growing food category due to both consumer demands and health claims by manufacturers. Probiotics are considered one of the best choices for meeting these demands. Traditionally, the food vehicle for introducing probiotics to consumers was dairy products, and to expand the benefits of probiotics for a wider range of consumers, the need to use other food items was essential. To achieve this goal while maximising the benefits of probiotics, protection methods used during food processing were tackled. The microencapsulation of probiotics is a promising methodology for achieving this function. This review highlights the use of the microencapsulation of probiotics in order to functionalise food items that initially were not considered suitable for probiotication, such as baked products, or to increase their functionality such as dairy products. The co-microencapsulation of probiotics with other functional ingredients such polyphenol, prebiotics, or omega-3 is also highlighted.

Encapsulated Bifidobacterium BB-12 addition in a concentrated lactose-free yogurt: Its survival during storage and effects on the product's properties.

This work aims to manufacture a new concentrated lactose-free probiotic yogurt. For this purpose, the probiotic Bifidocaterium BB-12 was incorporated in a concentrated lactose-free yogurt, both in its free form and previously encapsulated. Previous cell encapsulation was performed using the spray-drying technique with the following wall materials: lactose-free milk, lactose-free milk and inulin, and lactose-free milk and oligofructose. Thus, three different probiotic powders were obtained and added separately to three fractions of concentrated lactose-free yogurt. The probiotic survival of both powders and yogurts was evaluated during refrigerated storage. Likewise, the viability of starter cultures in yogurt (Lactobacillus bulgaricus and Streptococcus thermophilus) was controlled. In addition, the physicochemical properties of the four yogurts were also measured (color, pH and acidity, and texture properties). All three powders showed good probiotic viability (>8 log CFU g-1) throughout 120 days of storage at 4 °C. In turn, yogurt formulations (with the addition of powders or free bifidobacteria) presented probiotic viability above 7 log CFU g-1 after storage; as well as the starter cultures (>8 log UFC g-1). Yogurt with probiotic powder from lactose-free milk showed a more yellowish color; however, these differences would not be detected by the human eye (ΔE < 3.00). The yogurt with bifidobacteria free cells showed a greater post-acidification process (pH 4.18 to 4.02 and titratable acidity 1.52 to 1.89). It was not observed differences for firmness values of yogurt with free cells addition and yogurt with lactose-free milk and oligofructose powder addition. A slight significant decrease in the cohesiveness was observed in the yogurt elaborated with bifidobacteria free cells. The gumminess showed fluctuating values between all concentrated lactose-free yogurts. At the end of this study, we conclude that these probiotic powders can be incorporated into innovative lactose-free yogurts.

Teff-Based Probiotic Functional Beverage Fermented with Lactobacillus rhamnosus and Lactobacillus plantarum.

Consumers are demanding healthier foods, and the increasing drawbacks associated with dairy-based products have driven efforts to find plant-based probiotic alternatives. Consequently, this study aimed to evaluate the suitability of a teff-based substrate for delivering the potential probiotics, Lactobacillus rhamnosus GG (LGG) and Lactobacillus plantarum A6 (LA6) with a view to developing probiotic functional beverages. Single-strain and mixed-strain fermentations were performed without any pH control. In single-strain fermentation, LA6 grew to 8.157-8.349 log cfu/mL. Titratable acidity (TA) and pH were measured between 0.513-1.360 g/L and 4.25-3.91, respectively. The explored optimum variables were fermentation time (15 h) and inoculum (6 log cfu/mL). As a result of fermentation, maltose and glucose decreased, but lactic and acetic acids increased. In mixed-strain fermentation, LGG and LA6 were able to grow to 8.247 and 8.416 log cfu/mL, respectively. The pH, TA, lactic, and acetic acids varied between 6.31-3.92, 0.329-1.501 g/L, 0-1672 mg/L, and 20-231.5 mg/L, respectively. In both fermentations, microbial growth reached the stationary phase close to a pH of 4.21-4.82 while sugars were not consumed completely. Less than 5% ethanol was detected, which indicated a non-alcoholic beverage. A combination of the two evaluated lactobacilli strains reduced fermentation time. In conclusion, a substrate made of whole grain teff flour without any supplement could be used as a substrate to produce functional probiotic beverages.

Encapsulation of probiotics in soybean protein-based microparticles preserves viable cell concentration in foods all along the production and storage processes.

The influence on the stability of Lactobacillus plantarum CECT 220 (25 °C/60% relative humidity) of microencapsulation by simple coacervation followed by spray-drying using different Ca2+-to-soybean protein isolate ratios was evaluated. After optimisation, the selected soybean protein concentrate (SPC) microparticles were used to evaluate the tolerance of L. plantarum under acidic conditions (lactic acid, pH = 4; and HCl, pH = 3) and heat stress (80 °C for 1 min) in contrast to free cells. Moreover, after the heat treatment, the influence of the simulated gastric fluid was evaluated. Additionally, different foods were formulated using either microencapsulated or freeze-dried L. plantarum, and the stability of cells during the shelf-life of the formulated foods was studied. Results show that encapsulation with SPC enhanced significantly the stability of the Lactic Acid Bacteria all along the probiotic food value chain, from production to the end of the food shelf-life.

Isolation of Enterococcus faecium, characterization of its antimicrobial metabolites and viability in probiotic Minas Frescal cheese.

The aim of this study was to isolate Enterococcus faecium from raw milk samples, to characterize its antimicrobial metabolites, and to evaluate its viability in a probiotic Minas Frescal cheese. For this, antagonist activity against Listeria monocytogenes, safety aspects and biochemical, genotypic, and probiotic characteristics of the isolates were evaluated. Minas Frescal cheese was manufactured with the isolate that showed the best characteristics in vitro, and its viability in the product was evaluated. It was observed that of the 478 lactic acid bacteria isolates, only isolate E297 presented antagonist activity, genes encoding for enterocin production and absence of virulence factors. Besides that, E297 presented probiotic characteristics in vitro, and maintained its viability (8.09 log CFU mL-1) for 14 days of cold storage, when it was added to cheese. Therefore, isolate E297 can be considered a promising microorganism for the manufacture of probiotic foods, especially Minas Frescal cheese.

Biofabrication of Bacterial Constructs: New Three-Dimensional Biomaterials.

An enormous number of bacteria live in almost every environment; from deep oceans to below the surface of the earth or in our gastrointestinal tract. Although biofabrication is growing and maturing very quickly, the involvement of bacteria in this process has not been developed at a similar pace. From the development of a new generation of biomaterials to green bioremediation for the removal of hazardous environmental pollutants or to develop innovative food products in a recent trend, researchers have used cutting-edge biofabrication techniques to reveal the great potential of 3D structured bacterial constructs. These 3D bacterial workhouses may fundamentally change our approach toward biomaterials.

Quality of ready-to-eat swordfish fillets inoculated with Lactobacillus paracasei IMPC 2.1.

BACKGROUND: The quality of marinated ready-to-eat (RTE) swordfish fillets, with or without inoculation with the probiotic strain Lactobacillus paracasei IMPC 2.1, was assessed over 3 months of refrigerated storage at 4 °C. RTE probiotic and control fish fillets were sampled after 7, 14, 30, 60, and 90 days of storage. Microbiological tests were performed, and fatty acid (FA) profiles and malondialdehyde content were examined. Microbiological counts, including total viable count, lactic acid bacteria (LAB), yeasts, moulds, Enterobacteriaceae, and Pseudomonadaceae were determined. RESULTS: Inoculation successfully ensured the growth of the probiotic strain and prevented the growth of other LAB. The two RTE products showed significant differences in lipid profile and lipid oxidation during storage. In particular, inoculation with L. paracasei IMPC 2.1 increased the amount of polyunsaturated FAs and limited the amount of monounsaturated FAs and oleic acid, as well as lipid oxidation. It thus represents an interesting strategy for preserving the chemical quality of fish fillets and an alternative means of delivering probiotics. CONCLUSION: Probiotic inoculation with Lactobacillus paracasei seemed to delay lipid oxidation of the fish flesh and increase the retention of polyunsaturated FAs, suggesting a potential application for this strain in the seafood industry. © 2018 Society of Chemical Industry.

Suitability of a probiotic Lactobacillus paracasei strain as a starter culture in olive fermentation and development of the innovative patented product "probiotic table olives".

Probiotic bacteria are generally available for consumers as concentrated preparations or incorporated in milk-based foods. Due to an increased interest of the market for probiotic foods as well as to meet a demand of industry for innovation, a new kind of probiotic food has been developed using table olives as a carrier. Green table olives, produced according to the Spanish-style, are obtained by a fermentation which can be carried out by spontaneous microflora, even if the use of starter cultures is desirable to obtain a more controlled process. In this regard, the selected strain Lactobacillus paracasei IMPC 2.1 of human origin was used in the dual role of starter and probiotic culture, and here we describe the different aspects which have been evaluated and solved to utilize that strain for the development of a new table olive-based probiotic food. These aspects include selection of the strain on the basis of its probiotic properties, molecular characterization, compatibility with the carrier food, and efficacy as starter. The final product meets commercial and functional requirements throughout its shelf-life.

Efficacy of barley based probiotic food mixture in treatment of pathogenic E.coli induced diarrhoea in mice.

An indigenous food mixture was developed by mixing barley flour (raw and germinated), whey powder and tomato pulp in the ratio of 2:1:1:1 (w/w). The developed food (100 g) was mixed with water (500 ml) and autoclaved at 1.5 kg/cm(2) for 15 min. It was then cooled and inoculated with Lactobacillus acidophilus curd (5%) and incubated at 37°C for 12 h containing 10(6) cfu/ml broth. Fermented food mixture formulated from germinated barley flour maintained adequate cell viability (8.88 cfu/ml) as compared to non-germinated food mixture. To study the therapeutic effect of the food mixture, diarrhoea was induced in mice using 0.5 ml of aqueous suspension orally with the help of sterilized syringe to each of the overnight fasted mice and the mice were examined till onset of diarrhoea. The aqueous suspension was prepared by using 10 ml of six h old culture of E. coli cells (5 × 10(11) cfu/ml) and 6 ml alkaline solution (powdered chalk (40%), colloidal kaolin (43%) and magnesium trisilicate (17%) and both were mixed in 10:6 (v/v) proportions. After induction of diarrhoea, the mice were divided in two groups, control and experimental. The control group was fed on unfermented food mixture whereas experimental group was fed on fermented food mixture. Faecal ash, nitrogen, moisture and coliform count increased while faecal lactobacilli count decreased in mice having diarrhoea. In the experimental group, which was fed on fermented food mixture, normal values were reached within 7 days of feeding but no such changes were observed in control group which was fed on unfermented food mixture. Liver and kidney showed lesions due to E. coli infection were significantly alleviated on feeding of probiotic food mixture.