Avaliação da Sinergia da Atividade Antimicrobiana de Cepas de Bactérias Láticas Contra Patógenos

One of the most notable biological properties attributed to probiotics is their antimicrobial activity. Probiotics research has traditionally been focused on single strains, however, probiotic strains combinations can have a broader spectrum of action or synergistic effects, enhancing their protective action. The objective of this research was to evaluate the synergistic effect of lactic acid bacteria isolated from breast milk of different origins on the inhibition of bacterial pathogens of importance in human and animal medicine. Combinations of seven strains of lactic acidbacteria paired together were used and their inhibitory effect on bacterial pathogens Escherichia coli ATCC 25922, Salmonella enterica ATCC 13076, Klebsiella pneumoniae ATCC 13883, Acinetobacter baumannii ATCC 19606 and an enterohemorragic E. coli strain was investigated. The combination TUCO-5E + TUCO-L2 was the most effective in inhibiting the growth of all pathogens evaluated, highlighting its effect on K. pneumoniae y A. baumannii. Five of the evaluated combinations showed synergistic effects in the inhibition of extraintestinal pathogens: a) a synergistic effect was observed between strains TUCO-5E + TUCO-L2 for the inhibition of K. pneumoniae and A. baumannii, b) synergistic effect for TUCO-5E + TUCO-2, TUCO-5 + TUCO-2, and TUCO-3 + TUCO-17 on K. pneumoniae and, c) synergistic effect for TUCO-16 + TUCO-17 on A. baumannii. The combinations that showed synergy in the antimicrobial effect are good candidates for the development of multistrain probiotics for the prevention and treatment of specific bacterial infectionsUna de las propiedades biológicas más notables atribuidas a los probióticos es su actividad antimicrobiana. La investigación sobre probióticos se ha centrado tradicionalmente en cepas individuales,  sin embargo, las combinaciones de cepas probióticas pueden tener un espectro de acción más amplio o efectos sinérgicos, potenciando su acción protectora. El objetivo de esta investigación fue evaluar efecto sinérgico de cepas de bacterias lácticas aisladas de leche materna de diferentes orígenes sobre la inhibición de patógenos bacterianos de importancia en medicina humana y animal. Se emplearon combinaciones de siete cepas de bacterias lácticas juntadas en pares y se estudió su efecto inhibitorio sobre los patógenos bacterianos Escherichia coli ATCC 25922, Salmonella enterica ATCC 13076, Klebsiella pneumoniae ATCC 13883, y Acinetobacter baumannii ATCC 19606 y una cepa de E. coli enterohemorrágica. La combinación de las cepas TUCO-5E + TUCO-L2 fue la más efectiva en la inhibición del desarrollo de todos los patógenos evaluados, destacando su efecto sobre K. pneumoniae y A. baumannii. Cinco de las combinaciones evaluadas mostraron efectos sinérgicos en la inhibición de patógenos extraintestinales: a) se observó efecto sinérgico entre las cepas TUCO-5E + TUCO-L2 para la inhibición de K. pneumoniae y A. baumannii, b) efecto sinérgico para TUCO-5E + TUCO-2, TUCO-5 + TUCO-2, y TUCO-3 + TUCO-17 sobre K. pneumoniae y, c) efecto sinérgico para TUCO-16 + TUCO-17 sobre A. baumannii. Las combinaciones que mostraron sinergia en el efecto antimicrobiano son buenas candidatas para el desarrollo de probióticos multicepa para la prevención y el tratamiento de infecciones bacterianas específicasUma das propiedades biológicas mais notáveis atribuídas aos probióticos é a sua atividade antimicrobiana. A investigação sobre probióticos está focado tradicionalmente em cepas individuais, porém, as combinações de cepas probióticas podem ter um espectro de ação mais amplo, ou efeitos de sinergia que potenciam sua ação de proteção. O objetivo desta investigação foi avaliar o efeito de sinergia de cepas de bacterias láticas, isoladas do leite materno de diferentes origens, na inibição de patógenos bacterianos importantes na medicina humana e animal. Sete combinações de cepas de pares de bacterias láticas foram usadas e o efeito de inibição contra patógenos bacterianos Escherichia coli ATCC 25922, Salmonella enterica ATCC 13076, Klebsiella pneumoniae ATCC 13883, e Acinetobacter baumannii ATCC 19606 e uma cepa de E. coli enterohemorrágica. A combinação das cepas TUCO-5E + TUCO-L2 foi a mais efetiva na inibição do crescimento de todos os patógenos avaliados, destacando o efeito contra K. pneumoniae e A. baumannii. Cinco combinações apresentaram efeito de sinergia na inibição de patógenos extraintestinais: a) efeito de sinergia das cepas TUCO-5E + TUCO-L2 na inibição de K. pneumoniae e A. baumannii, b) efeito de sinergia das cepas TUCO-5E + TUCO-2, TUCO-5 + TUCO-2, e TUCO-3 + TUCO-17 contra K. pneumoniae e c) efeito de sinergia das cepas TUCO-16 + TUCO-17 contra A. baumannii. As combinações das cepas que apresentaram sinergia na inibição contra as cepas patógenas são boas candidatas para a elaboração de productos probióticos multicepa para prevenir e tratar infecções bacterianas específicas. baumannii, b) efeito de sinergia das cepas TUCO-5E + TUCO-2

Isolation, characterization, and immunomodulatory activity evaluation of probiotic strains from colostrum and canine milk

BackgroundThis study aimed to characterize potential probiotic strains for use in dogs to prevent infectious enteropathies. Lactic acid bacteria (LAB) isolated from canine milk and colostrum were characterized according to their functional properties, including their resistance to gastrointestinal conditions, inhibitory effect against pathogens, and intestinal adhesion.MethodsThe immunomodulatory effects of the strains were also analyzed in in vitro and in vivo studies. Among the strains evaluated, two LAB strains (TUCO-16 and TUCO-17) showed remarkable resistance to pH 3.0, bile salts, and pancreatin, as well as inhibitory effects against pathogenic Escherichia coli, Salmonella sp., and Clostridium perfringens.ResultsThe TUCO-16 and TUCO-17 strains induced a significant increase in the expression of TNF-α, IL-8, and TLR2 in canine macrophages. The oral administration of TUCO-16 and TUCO-17 strains to mice significantly augmented their resistance to pathogenic E. coli or Salmonella intestinal infections. Both canine strains reduced intestinal damage and pathogen counts in the liver and spleen and avoided their dissemination into the bloodstream. These protective effects were related to the ability of TUCO-16 and TUCO-17 strains to differentially modulate the production of IFN-γ, IFN-β, TNF-α, IL-6, KC, MCP-1, and IL-10 in the intestinal mucosa.ConclusionBoth strains, TUCO-16 and TUCO-17, are potential probiotic candidates for improving intestinal health in dogs, particularly for their ability to inhibit the growth of Gram-negative pathogens common in gastrointestinal infections and modulate the animal's immune response. Further studies are required to effectively demonstrate the beneficial effects of TUCO-16 and TUCO-17 strains in dogs

Isolation of lactic acid bacteria from swine milk and characterization of potential probiotic strains with antagonistic effects against swine-associated gastrointestinal pathogens

Probiotics are usually isolated from the gastrointestinal tract of humans and animals. The search of probiotics in human milk is a recent field of research, as the existence of the human milk microbiome was discovered only about a decade ago. To our knowledge, no reports regarding the potential probiotic effect of bacteria from swine milk were published. In this work, we isolated several lactic acid bacteria from swine milk and evaluated them for them potential as probiotics. Among the isolated strains, Lactobacillus curvatus TUCO-5E showed antagonistic effects against swine-associated gastrointestinal pathogens. TUCO-5E was able to reduce the growth of enterotoxigenic and enterohemorragic E. coli strains as well as pathogenic Salmonella. In vitro exclusion and displacement assays in intestinal epithelial cells showed a remarkable antagonistic effect for L. curvatus TUCO-5E against Salmonella TUCO-I7 and S. enterica ATCC 13096. Moreover, by using a mice model of Salmonella infection we were able to demonstrated that L. curvatus TUCO-5E preventive administration during 5 consecutive days was capable of decreasing the number of S. typhimurium in the liver and spleen of treated mice when compared to controls, and avoided dissemination of the pathogen to the blood stream. Then, we demonstrated here that swine milk is an interesting source for finding beneficial bacteria. In addition, the results of this work suggest that L. curvatus TUCO-5E is a good candidate for in vivo studying the protective effect of probiotics against intestinal infection and damage induced by Salmonella infection in the porcine host.On isole habituellement les probiotiques du tractus gastro-intestinal d’humains et d’animaux. La recherche de probiotiques dans du lait humain est un domaine de recherche récent, puisque la découverte du microbiome laitier humain ne date que d’environ 10 ans. À notre connaissance, on n’a jamais publié de rapport sur le possible effet probiotique de bactéries issues de lait porcin. Dans le présent ouvrage, nous avons isolé plusieurs bactéries lactiques du lait porcin et avons évalué leur potentiel probiotique. Parmi les souches isolées, Lactobacillus curvatus TUCO-5E a fait preuve d’antagonisme a` l’encontre de pathogènes gastro-intestinaux porcins. TUCO-5E a su diminuer la multiplication de souches entérotoxinogènes et entérohémorragiques d’Escherichia coli et de salmonelles pathogènes. Des expériences d’exclusion et de déplacement chez des cellules épithéliales intestinales ont permis de mettre en évidence une action antagoniste remarquable exercée par L. curvatus TUCO-5E envers Salmonella sp. souche TUCO-I7 et Salmonella enterica ATCC 13096. Par ailleurs, un modèle murin d’infection a` la salmonelle nous a permis de démontrer qu’une administration préventive de L. curvatus TUCO-5E pendant 5 jours consécutifs parvenait a` réduire le nombre de Salmonella enterica serovar Typhimurium dans le foie et la rate des souris traitées, comparativement aux témoins, et a` endiguer la propagation du pathogène dans la circulation sanguine. Dès lors, nous avons démontré que le lait porcin serait une source intéressante de bactéries bénéfiques. De plus, les résultats de cette étude laissent entendre que L. curvatus TUCO-5E pourrait faire l’objet d’études in vivo sur l’effet protecteur des probiotiques envers les infections intestinales et les dommages connexes occasionnés par les salmonelles dans l’hôte porcin. [Traduit par la Rédaction].Fil: Quilodrán Vega, Sandra Rayén. Universidad de Concepción. Facultad de Ciencias Veterinarias. Laboratorio de Microbiología de los Alimentos; ChileFil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucuman. Centro de Referencia Para Lactobacilos; ArgentinaFil: Valdebenito, José. Universidad de Concepción. Facultad de Ciencias Veterinarias. Laboratorio de Microbiología de los Alimentos; ChileFil: Salas, María José. Universidad de Concepción. Facultad de Ciencias Biológicas. Departamento de Microbiología. Laboratorio de Patogénesis Bacteriana; ChileFil: Parra, Cristian. Universidad de Concepción. Facultad de Ciencias Biológicas. Departamento de Microbiología. Laboratorio de Patogénesis Bacteriana; ChileFil: Ruiz, Alvaro. Universidad de Concepción. Facultad de Ciencias Veterinarias. Laboratorio de Microbiología de los Alimentos; ChileFil: Kitazawa, Haruki. Tohoku University. Graduate School of Agricultural Science. Laboratory of Animal Products Chemistry. Food and Feed Immunology Group; Japón. Tohoku University. Graduate School of Agricultural Science. International Education and Research Center for Food Agricultural Immunology. Livestock Immunology Unit ; JapónFil: García, Apolinaria. Univesidad de Concepción. Facultad de Ciencias Biológicas. Laboratorio de Patogénesis Bacteriana. Grupo de Investigación de Immunobiótica; Chil

Draft genome sequence of probiotic lactobacillus brevis TUCO-5E, isolated from porcine milk

This report describes the draft genome sequence of Lactobacillus brevisTUCO-5E, a probiotic strain isolated from porcine maternal milk. The reads were generated by a whole-genome sequencing (WGS) strategy on an Illumina MiSeq sequencer and were assembled into contigs with a total estimated size of 2,461,089 bp. A total of 2,455 open reading frames (ORFs) were predicted, including 2,301 protein-coding sequences. The draft genome sequence of L. brevis TUCO-5E will be useful for further studies of specific genetic features and for understanding the mechanisms of its probiotic properties in the porcine host.Fil: Quilodrán-Vega, Sandra Rayén. Universidad de Concepción. Facultad de Medicina Veterinaria. Departamento de Medicina Preventiva y Patología; ChileFil: Albarracín, Leonardo Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Hebert, Elvira Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Saavedra, Maria Lucila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Fonseca, Alexis. Universidad de Concepción. Departmento de Farmacología; ChileFil: Salas-Burgos, Alexis. Universidad de Concepción. Departmento de Farmacología; ChileFil: Kitazawa, Haruki. Tohoku University. Graduate School of Agricultural Science. Food and Feed Immunology Group; JapónFil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; Argentina. Universidad Nacional de Tucumán; Argentin