Characterization of lactic acid bacteria isolated from the poultry intestinal environment with anti-Salmonella activity in vitro.

The purpose of this research was the genotypic identification of lactic acid bacteria (LAB), isolated from the gastrointestinal tract (GIT) of healthy adult birds, and the study of their safety regarding antibiotic resistance, physiological and functional properties involved in the colonization of the GIT of poultry, and Salmonella exclusion, as members of a potential mixed probiotic supplement for poultry. The nucleotidic sequence from Lactobacillus crispatus P1, L. animalis L3, and Enterococcus faecium CRL 1385 (ex-J96) showed 100, 99.8, and 99.3% identity with L. crispatus DSM 20584 T, Ligilactobacillus salivarius ATCC 11741 T, and E. faecium ATCC 19434 T, respectively. These strains showed no resistance to relevant antibiotics usually administered to animals proposed by the European Food Safety Authority. They could endure the detrimental conditions of the gastrointestinal tract (pH 2.6 and oxgall 0.1 and 0.4% w/v). In an ex vivo assay, the LAB showed high adherence to the three sections of the GIT, reaching values higher than 70%. The adhesion to mucus was strain-dependent: L. crispatus CRL 1453 evidenced the highest adhesion (> 19%) while Lig. salivarius subsp. salivarius CRL 1417 and E. faecium CRL 1385 adhered to a lower extent (> 9 and 2%, respectively). Moreover, the LAB elicited remarkable anti-Salmonella activity, taking into account that they could inhibit elevated counts of different Salmonella serovars, especially the host-specific serovars S. Gallinarum and S. Pullorum (up to 8 log CFU/mL decrease in Salmonella counts).

Rapid Identification of new isolates of Acidipropionibacterium acidipropionici by fluorescence in situ hybridization (FISH)

Abstract Acidipropionibacterium acidipropionici is widely used for many applications, suchas propionic acid production, cereal silage, and also as probiotic. Due to this plethora of appli-cations, new isolates of A. acidipropionici with improved features are being searched for. Thesenew isolates must be accurately identified, however, most approaches become expensive andtime-consuming when the number of isolates is high. On the contrary, fluorescence in situhybridization allows the affordable, reliable, and rapid identification of microorganisms in purecultures and environmental and medical samples. Therefore, the aim of this work was to applya fluorescent in situ hybridization probe for the reliable identification of new A. acidipropioniciisolates. To this end, probe Pap446, specific for A. acidipropionici, was validated by hybridiza-tion assays with strains of this species from different origins, other species of the same genusor family, and unrelated genera. Eight isolates with propionibacterium characteristics wereobtained from milk and feces of cows. Probe Pap446, hybridized only with isolates III and VI.The identity of these isolates was further confirmed by PCR using group and species-specificprimers for propionibacteria and 16S rDNA sequencing.

Resumen Acidipropionibacterium acidipropionici es ampliamente usada para diversas aplicaciones, como producción de ácido propiónico, ensilado de cereales y probiótico. Debido a esta variedad de aplicaciones, continuamente se buscan nuevos aislamientos de A. acidipropionici con características nuevas. Estos nuevos aislamientos deben ser identificados correctamente, pero la mayoría de las técnicas disponibles resultan costosas e insumen mucho tiempo cuando el número de aislamientos es elevado. Por el contrario, la hibridación fluorescente in situ permite una identificación barata, confiable y rápida de microorganismos en cultivos puros y en muestras ambientales y médicas. Por lo tanto, el objetivo de este trabajo fue la aplicación de una sonda oligonucleotídica en un protocolo de hibridación fluorescente in situ para la identificación confiable de nuevos aislamientos de A. acidipropionici. Con este fin, se validó la sonda Pap446, específica de A. acidipropionici mediante ensayos de hibridación con cepas de esta especie de diferente origen, otras especies del mismo género o familia, y géneros no relacionados. Se obtuvieron ocho aislamientos con características de propionibacterias a partir de leche y heces de vacas. La sonda Pap446, hibridó únicamente con los aislamientos III y VI. La identidad de estos aislamientos fue confirmada a través de PCR con cebadores específicos para propionibacterias y para A. acidipropionici, y mediante secuenciación del ADNr 16S.

Rapid identification of new isolates of Acidipropionibacterium acidipropionici by fluorescence in situ hybridization (FISH).

Acidipropionibacterium acidipropionici is widely used for many applications, such as propionic acid production, cereal silage, and also as probiotic. Due to this plethora of applications, new isolates of A. acidipropionici with improved features are being searched for. These new isolates must be accurately identified, however, most approaches become expensive and time-consuming when the number of isolates is high. On the contrary, fluorescence in situ hybridization allows the affordable, reliable, and rapid identification of microorganisms in pure cultures and environmental and medical samples. Therefore, the aim of this work was to apply a fluorescent in situ hybridization probe for the reliable identification of new A. acidipropionici isolates. To this end, probe Pap446, specific for A. acidipropionici, was validated by hybridization assays with strains of this species from different origins, other species of the same genus or family, and unrelated genera. Eight isolates with propionibacterium characteristics were obtained from milk and feces of cows. Probe Pap446, hybridized only with isolates III and VI. The identity of these isolates was further confirmed by PCR using group and species-specific primers for propionibacteria and 16S rDNA sequencing.

Protection of the intestinal epithelium of poultry against deleterious effects of dietary lectins by a multi-strain bacterial supplement.

The intake of antinutritional factors produce impairment on the intestinal digestive function, impeding the efficient use of nutrients. Probiotics could be useful in poultry breeding to prevent negative effects of antinutritional factors, like the dietary lectins soybean agglutinin (SBA) and wheat germ agglutinin (WGA). Therefore, this investigation aimed to verify that SBA and wheat, which contains WGA, exert harmful effects on the intestinal mucosa and the digestive system of young poultry, and determine if the administration of probiotics able to capture lectins could counteract their effects. The trials performed demonstrated that a mixture of Bifidobacterium infantis CRL 1395, Enterococcus faecium LET 301, Lactobacillus salivarius LET 201, L. reuteri LET 210, and Propionibacterium acidipropionici LET 103, strains with ex vivo ability to interfere with the interaction of lectins and epithelial cells, has no negative effect on young chickens health. Middle levels of SBA, as well as wheat as a source of WGA, resulted in lower activities of intestinal and brush border enzymes and alterations in the integrity and morphological parameters of the chicks jejunal mucosa. The bacteria blend increased the activity of several digestive enzymes and the intestinal maturation marker alkaline phosphatase in birds fed with a conventional diet. Besides, it partially countered the deleterious effects of increased content of SBA, as well as the negative effect of a dietary source of WGA, on digestive enzymes activity and intestinal mucosa integrity. The results highlight the capability of multifunctional bacterial mixtures to protect the digestive system of avian against residual dietary lectins.