Effects of Bifidobacterium longum Subsp. infantis CECT 7210 and Lactobacillus rhamnosus HN001, Combined or Not With Oligofructose-Enriched Inulin, on Weaned Pigs Orally Challenged With Salmonella Typhimurium

Salmonella is a common causative agent of enteric disease and is developing mechanisms of resistance to antimicrobials. Probiotics, such as bifidobacteria and lactobacilli, and prebiotic fibers are a potential alternative to counteract this pathogen as they have demonstrated effectiveness in preventing its adhesion, reducing intestinal damage, and enhancing the host immune system. Furthermore, the benefits are expected to be potentiated when these compounds are administered together. A trial was performed to evaluate the efficacy of two probiotic strains (Bifidobacterium longum subsp. infantis CECT 7210 (Laboratorios Ordesa S.L.) and Lactobacillus rhamnosus HN001, combined or not with a prebiotic containing oligofructose-enriched inulin, against Salmonella Typhimurium. Ninety-six piglets (28 days old) were distributed into 32 pens assigned to 5 treatments: one non-challenged (control diet, CTR+) and four challenged: control diet (CTR−) or supplemented with probiotics (>3 × 10 10 cfu/kg each strain, PRO), prebiotic (5%, PRE), or their combination (SYN). After 1 week of adaptation, animals were orally challenged with Salmonella Typhimurium. Feed intake, weight, and clinical signs were recorded. On days 4 and 8 post-inoculation (PI), one animal per pen was euthanized, and samples from blood, digestive content, and ileal tissues were collected to determine Salmonella counts, fermentation products, ileal histomorphology, and serum TNF-α and Pig-MAP concentrations. The effect of the oral challenge was evidenced by animal performance, fecal consistency, and intestinal architecture. Regarding the experimental treatments, animals belonging to the PRO group experienced a faster clearance of the pathogen, with more pigs being negative to its excretion at the end of the study and recovering the impaired ileal villi/crypt ratio more rapidly. Animals receiving the PRE diet showed a lower intestinal colonization by Salmonella, with no countable levels (<3 cfu/g) in any of the analyzed samples, and an augmented immune response suggested by serum Pig-MAP concentrations. Treatments including the prebiotic (PRE and SYN) showed similar changes in the fermentation pattern, with an increase in the molar percentage of valeric acid concentration in the colon. The SYN group, however, did not show any of the outcomes registered for PRO and PRE in Salmonella colonization or in immunity markers, suggesting the lack of synbiotic action in this animal model. Further research is needed to better understand the complex mechanisms behind these effects

Aislamiento, identificación y caracterización de nuevos probióticos con propiedades funcionales para su aplicación en alimentación

La relación entre el hospedador y los microorganismos que colonizan el tracto gastrointestinal (GIT) es importante y fundamental para mantener un buen estado de salud, ya que se ha observado que la baja diversidad microbiana intestinal está asociada con el riesgo de enfermedades. Debido a esto, se han empleado diferentes técnicas para identificar y caracterizar bacterias con propiedades probióticas que puedan usarse como ingredientes en alimentos funcionales y de esta manera permitir el restablecimiento de las comunidades microbianas del GIT (aumento de la diversidad) o simplemente para mantener su buen funcionamiento. El principal objetivo de este trabajo ha sido la creación y caracterización de una colección de cepas probióticas, pertenecientes a los géneros Bifidobacterium y Lactobacillus, propiedad de Laboratorios Ordesa, a partir de muestras biológicas que incluyeron leche materna y heces de madre-bebé lactante. El uso de técnicas dependientes e independientes de cultivo, incluida la amplificación y secuenciación de un fragmento del gen 16S rARN y diferentes test bioquímicos (API 50CH y API ZYM) permitió identificar 288 cepas, donde Bifidobacterium fue el género predominante, representado por un 93.8% de todos los aislados, comparado con el género Lactobacillus (6,3%). Además de la identificación de las cepas aisladas, se realizaron análisis in vitro para evaluar diferentes propiedades funcionales, que incluyeron la capacidad de producción de ácidos conjugados a partir de ácido linoléico (LA) y ácido linolénico (LNA), la actividad antimicrobiana frente a tres patógenos relacionados con la mastitis en la lactancia y la producción de ácido gama-aminobutírico (GABA) a partir del Monoglutamato de Sodio (MGS). De acuerdo a los resultados obtenidos, se encontró que una cepa de Lactobacillus gasseri (ORD0529) fue capaz de inhibir el crecimiento de Staphylococcus aureus CECT4013, microorganismo causante de la mastitis en la lactancia. En cuanto a la producción de CLA y CLNA, se encontraron 10 cepas buenas productoras de estos ácidos grasos conjugados, siendo mayoritariamente bacterias del género Bifidobacterium y diferentes especies como Bifidobacterium longum subsp. longum (ORD0084), Bifidobacterium pseudocatenulatum (ORD0105), Bifidobacterium breve (ORD0123, ORD0124, ORD0128, ORD 0134, ORD0138 y ORD0294) y Bifidobacterium bifidum (ORD0202); y del género Lactobacillus solo se obtuvo una cepa perteneciente a Lactobacillus oris (ORD0255). Todas estas cepas fueron patentadas y depositadas en la Colección Española de Cultivos Tipo (CECT) bajo el número WO 2014/096352 A2, excepto la ORD0084 y ORD0105. Respecto a la producción de GABA, sólo se detectó la presencia de la enzima GAD en 7 cepas del género Bifidobacterium, concretamente en especies de Bifidobacterium adolescentis (ORD00106, ORD0119, ORD208, ORD0410, ORD0459 y ORD0465) y Bifidobacterium angulatum (ORD0026), sin embargo solo 6 de ellas fueron capaces de exportar el GABA. Los resultados obtenidos sugieren que estas cepas podrían ser tenidas en cuenta, para emplearse como ingredientes en productos alimentarios y/o complementos.The relationship between the human host and microorganisms that colonize the gut is important and fundamental to maintain a healthy balance and several research suggest that a gut microbiota with low diversity may have negative consequences for health. Because of that, different techniques have been employed to identify and characterize bacteria with probiotic properties that could be used as food ingredients. The aim of this work has been created an Ordesas´s strain collection, belonging to Bifidobacterium and Lactobacillus genera, from human milk and feces of breast feeding babies and their mothers. A total of 288 lactic acid bacteria from these samples were identified through several molecular biology techniques including partial amplification of 16S rRNA gene and also biochemical tests (API 50CH and API ZYM). Bifidobacterium was the most prevalent genus, representing 93,8% of the isolates followed by Lactobacillus (6,3%). Functional properties were evaluated by in vitro assays, including production of conjugated fatty acid from linoleic acid (LA) and linolenic acid (LNA), antagonistic activity against three different pathogens related with lactational mastitis and production of gamma-aminobutyric acid (GABA) from Monosodium Glutamate (MGS). According to the results, a strain of Lactobacillus oris (ORD0529) was able to inhibit the growth of Staphylococcus aureus CECT4013, microorganism that cause lactational mastitis. Regarding the production of CLA and CLNA, 10 strains were the best producers of these conjugated fatty acids, being mostly of the Bifidobacterium genus and different species such as Bifidobacterium longum subsp. longum (ORD0084), Bifidobacterium pseudocatenulatum (ORD0105), Bifidobacterium breve (ORD0123, ORD0124, ORD0128, ORD 0134, ORD0138 and ORD0294) and Bifidobacterium bifidum (ORD0202); and only a strain belonging to Lactobacillus oris (ORD0255) was obtained from the genus Lactobacillus. All of these strains were patented and deposited in the Spanish Type Culture Collection (ATTC), except the ORD0084 and ORD0105. Regarding the production of GABA, only some bacteria of the Bifidobacterium genus had the GAD enzyme and were able to export GABA, including species of Bifidobacterium adolescentis (ORD0106, ORD0119, ORD0410, ORD0459 and ORD0465) and Bifidobacterium angulatum (ORD0026). The GABA production was strain-dependent. The results obtained in this work provide novel opportunities for the development of functional foods

Aislamiento, identificación y caracterización de nuevos probióticos con propiedades funcionales para su aplicación en alimentación /

La relación entre el hospedador y los microorganismos que colonizan el tracto gastrointestinal (GIT) es importante y fundamental para mantener un buen estado de salud, ya que se ha observado que la baja diversidad microbiana intestinal está asociada con el riesgo de enfermedades. Debido a esto, se han empleado diferentes técnicas para identificar y caracterizar bacterias con propiedades probióticas que puedan usarse como ingredientes en alimentos funcionales y de esta manera permitir el restablecimiento de las comunidades microbianas del GIT (aumento de la diversidad) o simplemente para mantener su buen funcionamiento. El principal objetivo de este trabajo ha sido la creación y caracterización de una colección de cepas probióticas, pertenecientes a los géneros Bifidobacterium y Lactobacillus, propiedad de Laboratorios Ordesa, a partir de muestras biológicas que incluyeron leche materna y heces de madre-bebé lactante. El uso de técnicas dependientes e independientes de cultivo, incluida la amplificación y secuenciación de un fragmento del gen 16S rARN y diferentes test bioquímicos (API 50CH y API ZYM) permitió identificar 288 cepas, donde Bifidobacterium fue el género predominante, representado por un 93.8% de todos los aislados, comparado con el género Lactobacillus (6,3%). Además de la identificación de las cepas aisladas, se realizaron análisis in vitro para evaluar diferentes propiedades funcionales, que incluyeron la capacidad de producción de ácidos conjugados a partir de ácido linoléico (LA) y ácido linolénico (LNA), la actividad antimicrobiana frente a tres patógenos relacionados con la mastitis en la lactancia y la producción de ácido gama-aminobutírico (GABA) a partir del Monoglutamato de Sodio (MGS). De acuerdo a los resultados obtenidos, se encontró que una cepa de Lactobacillus gasseri (ORD0529) fue capaz de inhibir el crecimiento de Staphylococcus aureus CECT4013, microorganismo causante de la mastitis en la lactancia. En cuanto a la producción de CLA y CLNA, se encontraron 10 cepas buenas productoras de estos ácidos grasos conjugados, siendo mayoritariamente bacterias del género Bifidobacterium y diferentes especies como Bifidobacterium longum subsp. longum (ORD0084), Bifidobacterium pseudocatenulatum (ORD0105), Bifidobacterium breve (ORD0123, ORD0124, ORD0128, ORD 0134, ORD0138 y ORD0294) y Bifidobacterium bifidum (ORD0202); y del género Lactobacillus solo se obtuvo una cepa perteneciente a Lactobacillus oris (ORD0255). Todas estas cepas fueron patentadas y depositadas en la Colección Española de Cultivos Tipo (CECT) bajo el número WO 2014/096352 A2, excepto la ORD0084 y ORD0105. Respecto a la producción de GABA, sólo se detectó la presencia de la enzima GAD en 7 cepas del género Bifidobacterium, concretamente en especies de Bifidobacterium adolescentis (ORD00106, ORD0119, ORD208, ORD0410, ORD0459 y ORD0465) y Bifidobacterium angulatum (ORD0026), sin embargo solo 6 de ellas fueron capaces de exportar el GABA. Los resultados obtenidos sugieren que estas cepas podrían ser tenidas en cuenta, para emplearse como ingredientes en productos alimentarios y/o complementos.The relationship between the human host and microorganisms that colonize the gut is important and fundamental to maintain a healthy balance and several research suggest that a gut microbiota with low diversity may have negative consequences for health. Because of that, different techniques have been employed to identify and characterize bacteria with probiotic properties that could be used as food ingredients. The aim of this work has been created an Ordesas's strain collection, belonging to Bifidobacterium and Lactobacillus genera, from human milk and feces of breast feeding babies and their mothers. A total of 288 lactic acid bacteria from these samples were identified through several molecular biology techniques including partial amplification of 16S rRNA gene and also biochemical tests (API 50CH and API ZYM). Bifidobacterium was the most prevalent genus, representing 93,8% of the isolates followed by Lactobacillus (6,3%). Functional properties were evaluated by in vitro assays, including production of conjugated fatty acid from linoleic acid (LA) and linolenic acid (LNA), antagonistic activity against three different pathogens related with lactational mastitis and production of gamma-aminobutyric acid (GABA) from Monosodium Glutamate (MGS). According to the results, a strain of Lactobacillus oris (ORD0529) was able to inhibit the growth of Staphylococcus aureus CECT4013, microorganism that cause lactational mastitis. Regarding the production of CLA and CLNA, 10 strains were the best producers of these conjugated fatty acids, being mostly of the Bifidobacterium genus and different species such as Bifidobacterium longum subsp. longum (ORD0084), Bifidobacterium pseudocatenulatum (ORD0105), Bifidobacterium breve (ORD0123, ORD0124, ORD0128, ORD 0134, ORD0138 and ORD0294) and Bifidobacterium bifidum (ORD0202); and only a strain belonging to Lactobacillus oris (ORD0255) was obtained from the genus Lactobacillus. All of these strains were patented and deposited in the Spanish Type Culture Collection (ATTC), except the ORD0084 and ORD0105. Regarding the production of GABA, only some bacteria of the Bifidobacterium genus had the GAD enzyme and were able to export GABA, including species of Bifidobacterium adolescentis (ORD0106, ORD0119, ORD0410, ORD0459 and ORD0465) and Bifidobacterium angulatum (ORD0026). The GABA production was strain-dependent. The results obtained in this work provide novel opportunities for the development of functional foods

Assessment of the Effects of the Synbiotic Combination of Bifidobacterium longum subsp. infantis CECT 7210 and Oligofructose-Enriched Inulin Against Digestive Bacterial Infections in a Piglet Model

The use of bifidobacteria as probiotics has proven to be beneficial in gastroenteric infections. Furthermore, prebiotics such as inulin can enhance the survival and growth of these bacteria. Two trials were performed to evaluate the effects of the administration of Bifidobacterium longum subsp. infantis CECT 7210 and oligofructose-enriched inulin against Salmonella enterica serovar Typhimurium or enterotoxigenic Escherichia coli (ETEC) F4. A total of 72 (Salmonella trial) and 96 (ETEC F4 trial) weaned piglets were used in a 2 × 2 design (with or without synbiotic, inoculated or not with the pathogen). After adaptation, animals were orally inoculated. Performance and clinical signs were evaluated. On days 4 and 8 (Salmonella trial) and 3 and 7 (ETEC F4 trial) post-inoculation (PI), one animal per pen was euthanized. Blood, digestive content and tissue samples were collected and microbiological counts, fermentation products, serum inflammatory markers and ileum histomorphometry analysis were performed. Both challenges had an impact on faecal consistency (p < 0.001), including the faecal shedding of Salmonella and increased numbers of enterobacteria and coliforms. The synbiotic administration did not have any effect on pathogen loads but induced changes in the fermentation profile, such as increased valeric acid in both trials as well as decreased acetic acid, except for Salmonella -challenged animals. The effect on propionate varied among trials, increasing in challenged synbiotic-treated pigs and decreasing in non-challenged ones in the Salmonella trial (P interaction = 0.013), while the opposed occurred in the ETEC F4 trial (P interaction = 0.013). The administration of the synbiotic increased intraepithelial lymphocytes (IEL; p = 0.039) on day 8 PI in the Salmonella trial and a similar trend occurred in non-challenged pigs in the ETEC F4 trial (P interaction = 0.086). The results did not provide evidence of reduced pathogen load with the synbiotic, although a modulation in fermentative activity could be identified depending on the challenge. Consistent increases were found in IEL, suggesting that this synbiotic combination has some immunomodulatory properties

The Probiotic Combination of Bifidobacterium longum subsp. infantis CECT 7210 and Bifidobacterium animalis subsp. lactis BPL6 Reduces Pathogen Loads and Improves Gut Health of Weaned Piglets Orally Challenged with Salmonella Typhimurium

Probiotics have been demonstrated to be useful to enhance gut health and prevent gastrointestinal infections in humans. Additionally, some multi-strain probiotic combinations have been suggested to have greater efficacy than single strains. The objective of this study is to demonstrate the potential of a combination of the probiotic strains: Bifidobacterium longum subsp. infantis CECT 7210 (brand name B. infantis IM1®) and B. animalis subsp. lactis BPL6 to enhance gut health and to ameliorate the outcome of a Salmonella challenge using a weaning piglet model. Seventy-two 28-day-old weanling piglets, 7.7 (±0.28) kg of body-weight, were distributed in a 2 × 2 factorial design; treated or not with the probiotic combination and challenged or not with the pathogen. Animals were orally challenged after an adaptation period (Day 8) with a single dose (5 × 108 cfu) of Salmonella Typhimurium. One animal per pen was euthanized on Day 12 (Day 4 post-inoculation [PI]) and Day 16 (Day 8 PI). All parameters responded to the challenge and 4 deaths were registered, indicating a severe but self-limiting challenge. Improvements registered in the challenged animals due to the probiotic were: increased voluntary feed-intake (P probiotic × challenge = 0.078), reduced fecal excretion of Salmonella (P = 0.028 at Day 1 PI and P &lt; 0.10 at Days 3 and 5 PI), decreased rectal temperature (P probiotic × day = 0.048) and improvements in the villous:crypt ratio (P probiotic × challenge &lt; 0.001). Moreover, general probiotic benefits were observed in both challenged and non-challenged groups: decreased diarrhea scores of the PI period (P = 0.014), improved fermentation profiles on Day 8 PI (increased ileal acetic acid [P = 0.008] and a tendency to lower colonic ammonia concentrations [P = 0.078]), stimulation of intestinal immune response by increasing villous intraepithelial lymphocytes (P = 0.015 on Day 8 PI) and an improved villous:crypt ratio (P = 0.011). In conclusion, the multi-strain probiotic had a positive effect on reducing pathogen loads and alleviating animals in a Salmonella challenge. In addition, enhanced gut health and immunity was recorded in all animals receiving the probiotic, indicating an improvement in the post-weaning outcome

Effects of the Administration of Bifidobacterium longum subsp. infantis CECT 7210 and Lactobacillus rhamnosus HN001 and Their Synbiotic Combination With Galacto-Oligosaccharides Against Enterotoxigenic Escherichia coli F4 in an Early Weaned Piglet Model

We evaluated the potential of multi-strain probiotic (Bifidobacterium longum subsp. infantis CECT 7210 and Lactobacillus rhamnosus HN001) with or without galacto-oligosaccharides against enterotoxigenic Escherichia coli (ETEC) F4 infection in post-weaning pigs. Ninety-six piglets were distributed into 32 pens assigned to five treatments: one non-challenged (CTR+) and four challenged: control diet (CTR−), with probiotics (>3 × 10 10 CFU/kg body weight each, PRO), prebiotic (5%, PRE), or their combination (SYN). After 1 week, animals were orally inoculated with ETEC F4. Feed intake, weight, and clinical signs were recorded. On days 4 and 8 post-inoculation (PI), one animal per pen was euthanized and samples from blood, digesta, and tissues collected. Microbiological counts, ETEC F4 real-time PCR (qPCR) quantification, fermentation products, serum biomarkers, ileal histomorphometry, and genotype for mucin 4 (MUC4) polymorphism were determined. Animals in the PRO group had similar enterobacteria and coliform numbers to the CTR+ group, and the ETEC F4 prevalence, the number of mitotic cells at day 4 PI, and villus height at day 8 PI were between that observed in the CTR+ and CTR− groups. The PRO group exhibited reduced pig major acute-phase protein (Pig-MAP) levels on day 4 PI. The PRE diet group presented similar reductions in ETEC F4 and Pig-MAP, but there was no effect on microbial groups. The SYN group showed reduced fecal enterobacteria and coliform counts after the adaptation week but, after the inoculation, the SYN group showed lower performance and more animals with high ETEC F4 counts at day 8 PI. SYN treatment modified the colonic fermentation differently depending on the MUC4 polymorphism. These results confirm the potential of the probiotic strains and the prebiotic to fight ETEC F4, but do not show any synergy when administered together, at least in this animal model

ENGIU: Encuentro Nacional de Grupos de Investigación de UNIMINUTO.

El desarrollo del prototipo para el sistema de detección de Mina Antipersona (MAP), inicia desde el semillero ADSSOF perteneciente al programa de Administración en Seguridad y Salud en el trabajo de la UNIMINUTO, se realiza a partir de un detector de metales que emite una señal audible, que el usuario puede interpretar como aviso de presencia de un objeto metálico, en este caso una MAP. La señal audible se interpreta como un dato, como ese dato no es perceptible a 5 metros de distancia, se implementa el transmisor de Frecuencia Modulada FM por la facilidad de modulación y la escogencia de frecuencia de transmisión de acuerdo con las normas y resolución del Ministerio de Comunicaciones; de manera que esta sea la plataforma base para enviar los datos obtenidos a una frecuencia establecida. La idea es que el ser humano no explore zonas peligrosas y buscar la forma de crear un sistema que permita eliminar ese riesgo, por otro lado, buscar la facilidad de uso de elementos ya disponibles en el mercado