Molecular identification of Lactobacillus spp. associated with puba, a Brazilian fermented cassava food

Puba or carimã is a Brazilian staple food obtained by spontaneous submerged fermentation of cassava roots. A total of 116 lactobacilli and three cocci isolates from 20 commercial puba samples were recovered on de Man, Rogosa and Sharpe agar (MRS); they were characterized for their antagonistic activity against foodborne pathogens and identified taxonomically by classical and molecular methods. In all samples, lactic acid bacteria were recovered as the dominant microbiota (7.86 ± 0.41 log10 CFU/g). 16S-23S rRNA ARDRA pattern assigned 116 isolates to the Lactobacillus genus, represented by the species Lactobacillus fermentum (59 isolates), Lactobacillus delbrueckii (18 isolates), Lactobacillus casei (9 isolates), Lactobacillus reuteri (6 isolates), Lactobacillus brevis (3 isolates), Lactobacillus gasseri (2 isolates), Lactobacillus nagelii (1 isolate), and Lactobacillus plantarum group (18 isolates). recA gene-multiplex PCR analysis revealed that L. plantarum group isolates belonged to Lactobacillus plantarum (15 isolates) and Lactobacillus paraplantarum (3 isolates). Genomic diversity was investigated by molecular typing with rep (repetitive sequence)-based PCR using the primer ERIC2 (enterobacterial repetitive intergenic consensus). The Lactobacillus isolates exhibited genetic heterogeneity and species-specific fingerprint patterns. All the isolates showed antagonistic activity against the foodborne pathogenic bacteria tested. This antibacterial effect was attributed to acid production, except in the cases of three isolates that apparently produced bacteriocin-like inhibitory substances. This study provides the first insight into the genetic diversity of Lactobacillus spp. of puba.(AU)

Detection of CD4+ and CD8+ lymphocytes in the intestine of broiler chicks treated with Lactobacillus spp. and challenged with Salmonella enterica serovar Enteritidis

The expression of immune response as a leukocytic infiltrate by CD4+ and CD8+ cells in the epithelium and in the intestinal lamina propria of chicks fed Lactobacillus spp or cecal microflora (CM) and experimentally challenged or not with Salmonella enterica serovar Enteritidis (SE) was studied using immunohistochemistry. Three hundred and twenty day-of-hatch broiler chicks were divided into four groups of 80 birds each and orally received L. reuteri, L. salivarius, L. acidophilus, or CM. Each group was subdivided into four subgroups of 20 birds each, classified as follows: a subgroup did not receive any oral treatment (negative control), subgroup treated with L. spp or CM, subgroup treated with L. spp or CM and challenged with SE, and subgroup only challenged with SE (positive control). The results show that the oral treatment with L. reuteri, L. salivarius, L. acidophilus, or CM and challenge or not with SE stimulated bird immune response as determined by the leukocytic infiltrate by CD8+ lymphocytes followed by CD4+ in the epithelium and in the lamina propria of the duodenum, jejunum, and cecum of chicks up to 12 days of age. CD8+ lymphocyte number was significantly higher in the intestine of chicks receiving CM and challenged with SE. The duodenum, followed by the jejunum, were the segments in which the immune response, as shown by T, CD4+ and CD8+ cells, was stimulated with the greatest intensity.

Detection of CD4+ and CD8+ lymphocytes in the intestine of broiler chicks treated with Lactobacillus spp. and challenged with Salmonella enterica serovar Enteritidis

The expression of immune response as a leukocytic infiltrate by CD4+ and CD8+ cells in the epithelium and in the intestinal lamina propria of chicks fed Lactobacillus spp or cecal microflora (CM) and experimentally challenged or not with Salmonella enterica serovar Enteritidis (SE) was studied using immunohistochemistry. Three hundred and twenty day-of-hatch broiler chicks were divided into four groups of 80 birds each and orally received L. reuteri, L. salivarius, L. acidophilus, or CM. Each group was subdivided into four subgroups of 20 birds each, classified as follows: a subgroup did not receive any oral treatment (negative control), subgroup treated with L. spp or CM, subgroup treated with L. spp or CM and challenged with SE, and subgroup only challenged with SE (positive control). The results show that the oral treatment with L. reuteri, L. salivarius, L. acidophilus, or CM and challenge or not with SE stimulated bird immune response as determined by the leukocytic infiltrate by CD8+ lymphocytes followed by CD4+ in the epithelium and in the lamina propria of the duodenum, jejunum, and cecum of chicks up to 12 days of age. CD8+ lymphocyte number was significantly higher in the intestine of chicks receiving CM and challenged with SE. The duodenum, followed by the jejunum, were the segments in which the immune response, as shown by T, CD4+ and CD8+ cells, was stimulated with the greatest intensity.

Detecção de línfócitos T no intestino de frangos de corte tratados com lactobacillus spp. e desafiados com Salmonella enteritidis

O objetivo deste trabalho foi verificar por meio de imunoistoquímica se a inoculação oral de culturas de Lactobacillus acidophillus, L. reuteri, L. salivarius e a microbiota cecal de aves estimula a resposta imune celular no intestino de pintos de linhagem comercial, desafiadas ou não por Salmonella Enteritidis, na forma de infiltrado leucocitário por células T CD3+, CD4+ e CD8+. Foram utilizados 320 aves com um dia de vida, não sexados e livres de quimioterápicos. Cada grupo foi subdividido em quatro subgrupos com 20 aves cada e classificados em grupo que não recebeu tratamento (controle negativo), tratamento com L. spp ou MC, tratamento com L. spp ou MC e desafio com S. Enteritidis e tratamento com S. Enteritidis (controle positivo). Os resultados obtidos mostram que o tratamento com L. reuteri, L. salivarius, L. acidophillus ou microbiota cecal e o desafio com S. Enteritidis, determina resposta imune na forma de infiltrado leucocitário por linfócitos CD3+, CD4+ e CD8+ no epitélio e na lâmina própria do duodeno, jejuno e ceco de aves com até 12 dias de idade

The bacteria of the Lactobacillus sort have been very studied currently had to its important paper in the process of competitive exclusion where they contribute in the reduction of pathogenically genus, in special infections for Salmonella spp. it saw oral in the first days of life. These bacteria with probiotically characteristics provide beneficial effect to the general health of the host, through the stimulation and anti tumor modulation of the immune reply and properties, had probably the action on the cellular immune system. The objective of this work was to verify if Lactobacillus acidophillus, L. reuteri, L. salivarius and microbiota cecal of broiler chicken have been imunostimulated activities through its inoculation in birds of commercial ancestry with one day of life, defied or not for Salmonella Enteritidis, determining immune reply in the form of infiltrated leucocytes for cells T in the epithelium and intestinal proper blade.The gotten results demonstrate that the cultures of L. acidophillus, L. reuteri, L. salivarius and microbiota cecal defy or not with S. Enteritidis, they determine immune reply on the part of the host in the infiltrated form of leucocytes for lymphocytes CD3+, CD4+ and CD8+