Milk fermentation products of L. helveticus R389 activate calcineurin as a signal to promote gut mucosal immunity

<p/> <p>Background</p> <p>Fermented milks containing probiotic bacteria are a way of delivering bioactive constituents to targets in the gastrointestinal tract. We reported previously that the fermentation of milk at constant pH 6 by <it>L. helveticus </it>R389 increased its content of peptide fractions, and the oral administration of the non-bacterial fraction (FMSpH6) to mice increased total secretory IgA in the intestinal lumen and enhanced the number of IgA and various cytokines producing cells as well as the secretion of IL-6 by small intestine epithelial cells. We also demonstrated that this FMSpH6 was effective for the prevention of <it>Salmonella typhimurium </it>infection in mice. In this work, we studied in mice the impact of the oral administration of the supernatant of milk fermented by <it>L. helveticus </it>R389 on the gut physiology by measuring parameters such as calcium channels and E-cadherin expression, the activation of the biological signal calcineurin and mast and goblet cells, as a way to determine some mechanisms involved in the immunomodulating effects of the milk fermentation products, observed in previous studies. We analyzed the impact of the supernatant of milk fermented by <it>L. helveticus </it>R389 at pH6-controlled on the expression of calcineurin and on the reinforcement of the ephitelial barrier, measuring parameters such as calcium channels and E-cadherin expression and in the reinforcement of the non-specific immunity determining mast cells and goblet cells associated to the gut.</p> <p>Results</p> <p>We observed an enhanced expression of TRPV6 channels in the duodenum, indicating an improved capacity for dietary Ca2+ uptake. We demonstrated an enhanced expression of calcineurin in the small intestine, able to upregulate immune parameters such as IL-2 and TNF production, with an increase in the number of these cytokines secreting cells. We determined an increase in the number of mucosal mast cells and goblet cells, which would mean an improved state of mucosal surveillance at sites of infection.</p> <p>Conclusion</p> <p>The oral administration of the supernatant of milk fermented by <it>L. helveticus </it>R389 enhanced the gut mucosal immunity by improving the mechanisms that reinforce the epithelial and non-specific barriers and the gut functioning at sites of infection, with an improvement in the expression of the enzyme calcineurin, an important signal in the network that activates the gut immune system. The results of this work contribute to revealing the mechanisms underlying the immunomodulation of the gut immune function by fermented milks with probiotic bacteria.</p

Bacterias probióticas en productos lácteos fermentados

Las bacterias probióticas fueron definidas por la Organización Mundial de la Salud como microorganismos que, consumidos en adecuadas dosis, ejercen efectos benéficos sobre la salud del consumidor. La mayoría de estos microorganismos pertenecen a los géneros Lactobacillus y Bifidobacterium, son de origen intestinal (aislados de individuos sanos) y son incluidos en productos lácteos fermentados, principalmente yogures, para su llegada al consumidor. Su agregado a matrices alimenticias presenta numerosos desafíos tecnológicos, como lo son el mantenimiento de su viabilidad y funcionalidad a lo largo del proceso y conservación del alimento, como también el recuento selectivo o diferencial en medios de cultivos, lo que permite realizar el control de calidad del nivel de células viables, requisito fundamental para ejercer su actividad probiótica o benéfica para la salud. Desde mediados de los años 90, la Argentina, en sintonía con una tendencia mundial, se sumó al desarrollo comercial de leches fermentadas y, en menor medida, de quesos frescos conteniendo cultivos de L. acidophilus, L. casei y Bifidobacterium. Nuestro país fue además pionero (primer país de Latinoamérica en 1999) en el desarrollo comercial de quesos probióticos. El consumo regular de este tipo de alimentos permite mantener un nivel más saludable de vida, disminuyendo en cierto grado la incidencia, severidad y frecuencia de algunas patologías intestinales (diarreas, inflamación) o no intestinales (enfermedades respiratorias, alergias) o modulando de forma benéfica el tránsito intestinal o la actividad de la microbiota residente en el tracto gastrointestinal.Fil: Vinderola, Celso Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; Argentin

Cell Viability and Functionality of Probiotic Bacteria in Dairy Products

Probiotic bacteria, according to the definition adopted by the World Health Organization in 2002, are live microorganisms, which when administered in adequate amounts confer a health benefit to the host. Recent studies show that the same probiotic strain produced and/or preserved under different storage conditions, may present different responses regarding their susceptibility to the adverse conditions of the gastrointestinal tract, its capacity to adhere to the intestinal epithelium, or its immunomodulating capacity, the functionality being affected without changes in cell viability. This could imply that the control of cell viability is not always enough to guarantee the functionality (probiotic capacity) of a strain. Therefore, a new challenge arises for food technologists and microbiologists when it comes to designing and monitoring probiotic food: to be able to monitor the functionality of a probiotic microorganism throughout all the stages the strain goes through from the moment it is produced and included in the food vehicle, until the moment of consumption. Conventional methodological tools or others still to be developed must be used. The application of cell membrane functionality markers, the use of tests of resistance to intestinal barriers, the study of surface properties and the application of in vivo models come together as complementary tools to assess the actual capacity of a probiotic organism in a specific food, to exert functional effects regardless of the number of viable cells present at the moment of consumption

Infant formula supplemented with biotics: Current knowledge and future perspectives

Breastfeeding is natural and the optimal basis of infant nutrition and development, with many benefits for maternal health. Human milk is a dynamic fluid fulfilling an infant’s specific nutritional requirements and guiding the growth, developmental, and physiological processes of the infant. Human milk is considered unique in composition, and it is influenced by several factors, such as maternal diet and health, body composition, and geographic region. Human milk stands as a model for infant formula providing nutritional solutions for infants not able to receive enough mother’s milk. Infant formulas aim to mimic the composition and functionality of human milk by providing ingredients reflecting those of the latest human milk insights, such as oligosaccharides, bacteria, and bacterial metabolites. The objective of this narrative review is to discuss the most recent developments in infant formula with a special focus on human milk oligosaccharides and postbiotics.Fil: Salminen, Seppo. University of Turku. Faculty of Medicine; FinlandiaFil: Stahl, Bernd. Nutricia Research; Países Bajos. Utrecht Institute Of Pharmaceutical Sciences; Países BajosFil: Vinderola, Celso Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; ArgentinaFil: Szajewska, Hania. Medical University Of Warsaw; Poloni

Potential contribution of beneficial microbes to face the COVID-19 pandemic

The year 2020 will be remembered by a never before seen, at least by our generation, global pandemic of COVID-19. While a desperate search for effective vaccines or drug therapies is on the run, nutritional strategies to promote immunity against SARS-CoV-2, are being discussed. Certain fermented foods and probiotics may deliver viable microbes with the potential to promote gut immunity. Prebiotics, on their side, may enhance gut immunity by selectively stimulating certain resident microbes in the gut. Different levels of evidence support the use of fermented foods, probiotics and prebiotics to promote gut and lungs immunity. Without being a promise of efficacy against COVID-19, incorporating them into the diet may help to low down gut inflammation and to enhance mucosal immunity, to possibly better face the infection by contributing to diminishing the severity or the duration of infection episodes.Fil: Antunes, Adriane E.C.. Universidade Estadual de Campinas; BrasilFil: Vinderola, Celso Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; ArgentinaFil: Xavier-Santos, Douglas. Universidade Estadual de Campinas; BrasilFil: Sivieri, Katia. Universidade Estadual Paulista Julio de Mesquita Filho; Brasi

The role of homofermentative and heterofermentative lactic acid bacteria for alfalfa silage: a meta-analysis

Lactic acid bacteria (LAB) are usually employed as alfalfa silage inoculants to obtain high-quality feed for animal husbandry. However, the effects of these inoculants are still unclear and need to be studied extensively. Therefore, the objective of this meta-analysis was to quantitatively summarize published research studies that assess the effects of homofermentative (HoLAB) and heterofermentative lactic acid bacteria (HeLAB) on fermentation parameters, nutritive value, microbiological composition and aerobic stability of alfalfa silage. PubMed, ScienceDirect and Scopus have been screened for articles published from 1980 to 2018. The criteria for inclusion were: randomized and controlled trials using alfalfa silage and published in peer-reviewed journals. It was found that inoculation with LAB decreased silage pH, neutral detergent fibre, acid detergent fibre and ammoniacal nitrogen, while it increased dry matter and crude protein compared to control in the pooled raw mean difference random-effect model. Additionally, LAB inoculation decreased acetate, propionate, ethanol and butyrate concentrations, whereas it increased lactate. In addition, inoculants reduced the counts of yeasts and moulds. Lastly, LAB inoculation improved aerobic stability. To the best of our knowledge, this is the first meta-analysis that aims at comparing the application of HoLAB and HeLAB for alfalfa silage. In the pool estimate, positive effects attributable to the application of microbial silage inoculants were found in most of the evaluated parameters; supporting the importance of applying both types of inoculants to improve forage preservation for the livestock industry.EEA RafaelaFil: Blajman, Jesica E. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Blajman, Jesica E. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; ArgentinaFil: Vinderola, Celso Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Lactologia Industrial; ArgentinaFil: Vinderola, Celso Gabriel. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactologia Industrial; ArgentinaFil: Paez, Roxana Beatriz. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; ArgentinaFil: Signorini, Marcelo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; ArgentinaFil: Signorini, Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Hemolytic uremic syndrome and yoghurt: Popular belief and scientific evidence

Some health professionals discourage yogurt because of the risk of Hemolytic Uremic Syndrome (HUS), a serious disease caused by strains of Shiga toxin-producing E. coli (STEC). These bacteria can pass from the intestine of cattle to meat or milk under inadequate working conditions in slaughterhouses or milking plants. Undercooked hamburgers the main is vector of disease and unpasteurized raw milk or dairy products made with it, are another risk factors. In the dairy industry, yoghurt is made from milk that undergoes a double heat treatment. There are no reports of the presence of STEC in industrial yogurts in the modern bibliography, and reviews and meta-analysis do not point to yogurt as a risk factor for STEC, but rather unpasteurized milk. In this context, and given the scientific evidence currently available regarding STEC, HUS and yogurt, it would seem that we are in the presence of a spurious correlation, the association between two facts that have no causal relationship between them, rather than a scientific fact for which one (yogurt) may be responsible for the other (HUS).Algunos profesionales de la salud desaconsejan el consumo del yogur por el riesgo de provocar Síndrome Urémico Hemolítico, una enfermedad grave causada por cepas de E. coli productor de toxina Shiga (STEC por sus siglas en inglés). Estas bacterias pueden pasar del intestino del ganado vacuno a la carne o a la leche en condiciones inadecuadas de trabajo en frigoríficos o establecimientos productores de leche, respectivamente, siendo las hamburguesas insu-ficientemente cocidas el principal vector de la enfermedad y la leche cruda sin pasteurizar o los productos lácteos elaborados con ésta, otro factor de riesgo. En la industria láctea, el yogur se elabora con leche que es sometida a un doble tratamiento térmico. En la bibliografía moderna reportes de la presencia de STEC en yogures industriales, y los trabajos de revisión y meta-análisis no incluyen al yogur, pero sí a la leche sin pasteurizar, como vectores de trasmisión de STEC. En este contexto, y dada la evidencia científica disponible actualmente en relación a E. coli pro-ductor de toxina Shiga, el SUH y el yogur, parecería que estamos ante la presencia de una correlación espuria, la asociación de dos hechos que no tienen relación causal entre sí, más que a un hecho científico del cual uno (el yogur) es el responsable del otro (SUH).Palabras claves: E. coli, Hamburguesas, STEC, Síndrome Urémico Hemolítico, Yogures.Fil: Vinderola, Celso Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; ArgentinaFil: Rivas, Marta. Instituto Nacional de Enfermedades Infecciosas; Argentin

The Concept of Postbiotics

The scientific community has proposed terms such as non-viable probiotics, paraprobiotics, ghostbiotics, heat-inactivated probiotics or, most commonly, postbiotics, to refer to inanimate microorganisms and/or their components that confer health benefits. This article addresses the various characteristics of different definitions of 'postbiotics' that have emerged over past years. In 2021, the International Scientific Association for Probiotics and Prebiotics (ISAPP) defined a postbiotic as "a preparation of inanimate microorganisms and/or their components that confers a health benefit on the host". This definition of postbiotic requires that the whole or components of inactivated microbes be present, with or without metabolic end products. The definition proposed by ISAPP is comprehensive enough to allow the development of postbiotics from different microorganisms, to be applied in different body sites, encouraging innovation in a promising area for any regulatory category and for companion or production animals, and plant or human health. From a technological perspective, probiotic products may contain inanimate microorganisms, which have the potential to impart a health benefit. However, their contribution to health in most cases has not been established, even if at least one probiotic has been shown to confer the same health benefit by live or inanimate cells

Potential of Lactic Acid Bacteria Isolated From Different Forages as Silage Inoculants for Improving Fermentation Quality and Aerobic Stability

We aimed at isolating lactic acid bacteria (LAB) from different plant materials to study their crossed-fermentation capacity in silos and to find strains able to confer enhanced aerobic stability to silage. A total of 129 LAB isolates were obtained from lucerne (alfalfa), maize, sorghum, ryegrass, rice, barley, canola, Gatton panic, Melilotus albus, soy, white clover, wheat, sunflower, oat, and moha. Four Lactiplantibacillus plantarum subsp. plantarum strains (isolated from oat, lucerne, sorghum, or maize) were selected for their growth capacity. Identity (16S sequencing) and diversity (RAPD-PCR) were confirmed. Fermentative capacity (inoculated at 104, 105, 106, 107 CFU/g) was studied in maize silage and their cross-fermentation capacity was assessed in oat, lucerne, sorghum, and maize. Heterofermentative strains with the highest acetic acid production capacity conferred higher aerobic stability to maize silages. Regardless the source of isolation, L. plantarum strains, inoculated at a rate of 106 CFU/g, were effective to produce silage from different plant materials. From more than 100 isolates obtained, the application of a succession of experiments allowed us to narrow down the number of potential candidates of silage inoculants to two strains. Based on the studies made, L. plantarum LpM15 and Limosilactobacillus fermentum LfM1 showed potential to be used as inoculants, however further studies are needed to determine their performance when inoculated together. The former because it positively influenced different quality parameters in oat, lucerne, sorghum, and maize silage, and the latter because of its capacity to confer enhanced aerobic stability to maize silage. The rest of the strains constitute a valuable collection of autochthonous strains that will be further studied in the future for new applications in animal or human foods.EEA RafaelaFil: Puntillo, Melisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; ArgentinaFil: Gaggiotti, Mónica del Carmen. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Laboratorio de Calidad de Leche y Agroindustria; ArgentinaFil: Oteiza, Juan Martín. Centro de Investigación y Asistencia Técnica a la Industria. Laboratorio de Microbiología de los Alimentos (Neuquén); ArgentinaFil: Binetti, A. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Lactología Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactología Industrial; ArgentinaFil: Massera, Ariel Fernando. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Laboratorio de Calidad de Leche y Agroindustria; ArgentinaFil: Vinderola, Celso Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Lactologia Industrial. Universidad Nacional del Litoral. Facultad de Ingeniería Química. Instituto de Lactologia Industrial; Argentin

Cell counting and bacterial inoculum standardization by spectrophotometric method for Bifidobacterium animalis ssp. Lactis INL1 / Contagem de células e padronização de inóculos bacterianos pelo método espectrofotométrico para Bifidobacterium animalis ssp. Lactis INL1

The genus Bifidobacterium is present in the human intestinal microbiota and in abundance in breast milk, conferring probiotic features to this food. However, a large number of variables affect its growth, hindering possible analysis and its use in organic assays. Different direct and indirect methodologies of bacterial cell counting provide results in some units of measurement, such as colony-forming units (CFU) per gram or mL, or even cell number per mL. Standardizing bacterial inoculum used in long-term daily exposure experiments is sometimes difficult due to the numerous variables involved in plaques cultivation and incubation. Considering that, this study sought to standardize adjusted bacterial inoculum from the indirect method of bacterial counting by spectrophotometry, using calibration curve and plating confirmation to ensure the amount of cells to be inoculated. As a result, we found that colony-forming units grew at a 108 rate at 600 nm, within the 0.010–0.006 absorbance range, and that such rate remains regardless of differences within the CFU/mL count. This method can be used to adjust Bifidobacterium animallis ssp Lactis INL1 inoculum, considering its probiotic effect.