Protective Effect of Lactobacillus diolivorans 1Z, Isolated From Brazilian Kefir, Against Salmonella enterica Serovar Typhimurium in Experimental Murine Models.

Kefir is a beverage obtained by fermentation of milk or sugar solution by lactic acid bacteria and yeasts, and several health benefits have been attributed to its ingestion, part of them being attributed to Lactobacillus species. The objective of the present study was to evaluate, in vivo, the probiotic potential of Lactobacillus diolivorans 1Z, isolated from Brazilian kefir grains. Initially, conventional mice were orally treated daily or not during 10 days with a suspension of L. diolivorans 1Z, and then orally challenged with Salmonella enterica serovar Typhimurium. Treatment with L. diolivorans 1Z resulted in higher survival (70%) of animals after the challenge with the pathogen than for not treated mice (0%). When germ-free mice were monoassociated (GN-PS group) or not (GN-CS group) with L. diolivorans 1Z and challenged after 7 days with S. Typhimurium, Salmonella fecal counts were significantly lower (P < 0.05) for the GN-PS group when compared to the GN-CS group. Histopathological analysis revealed less damage to the ileum mucosa, as demonstrated by smallest perimeter of major lesions for mice of the GN-PS group in comparison to the group GN-CS (P < 0.05). These findings were accompanied by a lower expression of IFN-γ and TNF-α in the intestinal tissue of GN-PS mice. Additionally, translocation of S. Typhimurium to liver was significantly lower in GN-PS than in GN-CS mice (P < 0.05), and IgA levels in intestinal content and number of Kupffer cells in liver were higher. No difference was observed for hepatic cellularity between GN-PS and GN-CS groups (P > 0.05), but the pattern of inflammatory cells present in the liver was predominantly of polymorphonuclear in GN-CS group and of mononuclear in the GN-PS group, and a higher hepatic expression of IL-10 and TGF-ß was observed in GN-PS group. Concluding, L. diolivorans 1Z showed to be a potential probiotic strain that protected mice from death after challenge with S. Typhimurium, apparently by immunological modulation.

Antagonistic and protective effects against Salmonella enterica serovar typhimurium by Lactobacillus murinus in the digestive tract of gnotobiotic mice

In the present study, production of antagonistic substances by Lactobacillus murinus against enteropathogenic bacteria was evaluated in vivo as well as a possible protective effect against an oral challenge with Salmonella Typhimurium using a gnotobiotic animal model. A higher mean survival time (P 0.05) was observed in L. murinus-associated animals (7.89 ± 3.83 days) when compared with the control ones (4.44 ± 0.73 days). Lactobacillus murinus exerted a potent antagonistic effect in vivo against S. sonnei and to a lesser extent against S. Typhimurium as revealed by inhibitory halos around the feces of Lactobacillus-associated animals. Diameters of inhibitory halos around intestinal contents increased along the intestine tract, following proportionally the L. murinus population levels in the respective intestinal portions. Concluding, the present study showed that L. murinus association in gnotobiotic mice delayed the death after an oral challenge with S. Typhimurium and that inhibitory diffusible compounds obtained in in vitro assays were also produced in vivo and may be responsible for this effect.

No presente estudo, a produção de substâncias antagonistas por Lactobacillus murinus contra bactérias enteropatogênicas foi avaliada in vivo assim como um possível efeito protetor contra um desafio oral com Salmonella Typhimurium utilizando um modelo animal gnotobiótico. Um maior tempo médio de sobrevida (P 0,05) foi observado nos animais associados com L. murinus (7,89 ± 3,83 dias) quando comparado com os controles (4,44 ± 0,73 dias). Lactobacillus murinus exerceu um potente efeito antagonista in vivo contra S. sonnei e com menos intensidade contra S. Typhimurium como revelado pelos halos de inibição ao redor das fezes dos animais associados com L. murinus. Os diâmetros dos halos de inibição ao redor dos conteúdos intestinais aumentaram ao longo do trato digestivo, seguindo proporcionalmente os níveis populacionais de L. murinus nas respectivas porções intestinais. Concluindo, o presente estudo mostra que a associação com L. murinus em camundongos gnotobióticos retarda a morte após um desafio oral com S. Typhimurium e que compostos inibitórios difusíveis obtidos em ensaios in vitro foram também produzidos in vivo e podem ser responsáveis por este efeito.

Antagonistic and protective effects against Salmonella enterica serovar typhimurium by Lactobacillus murinus in the digestive tract of gnotobiotic mice

In the present study, production of antagonistic substances by Lactobacillus murinus against enteropathogenic bacteria was evaluated in vivo as well as a possible protective effect against an oral challenge with Salmonella Typhimurium using a gnotobiotic animal model. A higher mean survival time (P 0.05) was observed in L. murinus-associated animals (7.89 ± 3.83 days) when compared with the control ones (4.44 ± 0.73 days). Lactobacillus murinus exerted a potent antagonistic effect in vivo against S. sonnei and to a lesser extent against S. Typhimurium as revealed by inhibitory halos around the feces of Lactobacillus-associated animals. Diameters of inhibitory halos around intestinal contents increased along the intestine tract, following proportionally the L. murinus population levels in the respective intestinal portions. Concluding, the present study showed that L. murinus association in gnotobiotic mice delayed the death after an oral challenge with S. Typhimurium and that inhibitory diffusible compounds obtained in in vitro assays were also produced in vivo and may be responsible for this effect.

No presente estudo, a produção de substâncias antagonistas por Lactobacillus murinus contra bactérias enteropatogênicas foi avaliada in vivo assim como um possível efeito protetor contra um desafio oral com Salmonella Typhimurium utilizando um modelo animal gnotobiótico. Um maior tempo médio de sobrevida (P 0,05) foi observado nos animais associados com L. murinus (7,89 ± 3,83 dias) quando comparado com os controles (4,44 ± 0,73 dias). Lactobacillus murinus exerceu um potente efeito antagonista in vivo contra S. sonnei e com menos intensidade contra S. Typhimurium como revelado pelos halos de inibição ao redor das fezes dos animais associados com L. murinus. Os diâmetros dos halos de inibição ao redor dos conteúdos intestinais aumentaram ao longo do trato digestivo, seguindo proporcionalmente os níveis populacionais de L. murinus nas respectivas porções intestinais. Concluindo, o presente estudo mostra que a associação com L. murinus em camundongos gnotobióticos retarda a morte após um desafio oral com S. Typhimurium e que compostos inibitórios difusíveis obtidos em ensaios in vitro foram também produzidos in vivo e podem ser responsáveis por este efeito.

Antagonistic and protective effects against Salmonella enterica serovar typhimurium by Lactobacillus murinus in the digestive tract of gnotobiotic mice

In the present study, production of antagonistic substances by Lactobacillus murinus against enteropathogenic bacteria was evaluated in vivo as well as a possible protective effect against an oral challenge with Salmonella Typhimurium using a gnotobiotic animal model. A higher mean survival time (P 0.05) was observed in L. murinus-associated animals (7.89 ± 3.83 days) when compared with the control ones (4.44 ± 0.73 days). Lactobacillus murinus exerted a potent antagonistic effect in vivo against S. sonnei and to a lesser extent against S. Typhimurium as revealed by inhibitory halos around the feces of Lactobacillus-associated animals. Diameters of inhibitory halos around intestinal contents increased along the intestine tract, following proportionally the L. murinus population levels in the respective intestinal portions. Concluding, the present study showed that L. murinus association in gnotobiotic mice delayed the death after an oral challenge with S. Typhimurium and that inhibitory diffusible compounds obtained in in vitro assays were also produced in vivo and may be responsible for this effect.

No presente estudo, a produção de substâncias antagonistas por Lactobacillus murinus contra bactérias enteropatogênicas foi avaliada in vivo assim como um possível efeito protetor contra um desafio oral com Salmonella Typhimurium utilizando um modelo animal gnotobiótico. Um maior tempo médio de sobrevida (P 0,05) foi observado nos animais associados com L. murinus (7,89 ± 3,83 dias) quando comparado com os controles (4,44 ± 0,73 dias). Lactobacillus murinus exerceu um potente efeito antagonista in vivo contra S. sonnei e com menos intensidade contra S. Typhimurium como revelado pelos halos de inibição ao redor das fezes dos animais associados com L. murinus. Os diâmetros dos halos de inibição ao redor dos conteúdos intestinais aumentaram ao longo do trato digestivo, seguindo proporcionalmente os níveis populacionais de L. murinus nas respectivas porções intestinais. Concluindo, o presente estudo mostra que a associação com L. murinus em camundongos gnotobióticos retarda a morte após um desafio oral com S. Typhimurium e que compostos inibitórios difusíveis obtidos em ensaios in vitro foram também produzidos in vivo e podem ser responsáveis por este efeito.

Protection by Lactobacillus acidophilus UFV-H2B20 against experimental oral infection with Salmonella enterica subsp. enterica Ser. Typhimurium in gnotobiotic and conventional mice

The ability of Lactobacillus acidophilus UFV-H2B20 to antagonize Salmonella enterica subsp. enterica ser. Typhimurium and to reduce the pathological consequences for the host was determining using conventional and gnotobiotic animals. Conventional NIH mice received daily by gavage a 0.1 ml suspension containing about 10(8) cfu L. acidophilus UFV-H2B20 and germfree animals received a single 0.1 ml dose. The gnotobiotic and conventional groups were infected orally with 10² and 10(5) cfu of S. Typhimurium, respectively, 7 days after the beginning of treatment. Control groups were treated with sterile saline instead of Lactobacillus. Survival data showed a protective effect against the pathogenic bacteria in both conventional and gnotobiotic Lactobacillus-treated mice. L. acidophilus UFV-H2B20 colonized the digestive tract of gnotobiotic mice and the number of viable cells ranged from 10(9) to 10(10) cfu/g of faeces. In both experimental and control gnotobiotic animals, S. Typhimurium became rapidly established at a level ranging from 10(8) to 10(10) cfu/g of faeces and remained at high levels until the animals died or were sacrificed. In conclusion, the previous treatment of mice with L. acidophilus UFV-H2B20 protects the animals against the experimental infection with S. Typhimurium but this protection was not due to the reduction of the pathogenic populations in the intestines.

A capacidade de Lactobacillus acidophilus UFV-H2B20 de antagonizar Salmonella enterica subsp. enterica ser. Typhimurium, e de reduzir as conseqüências patológicas para o hospedeiro foram determinadas em animais convencionais e gnotobióticos. Camundongos NIH convencionais receberam diariamente, por via oral, 0,1 ml de uma suspensão contendo em torno de 10(8) ufc de L. acidophilus UFV-H2B20 e os animais sem germes receberam uma única dose de 0,1 ml. Os grupos gnotobióticos e convencionais foram desafiados oralmente com, respectivamente, 10² e 10(5) ufc de S. Typhimurium 7 dias após o início do tratamento. Os grupos controles foram tratados com salina estéril em vez do Lactobacillus. Dados de sobrevida mostraram um efeito protetor contra a bactéria patogênica em ambos os grupos convencional e gnotobiótico tratados com o Lactobacillus. L. acidophilus UFV-H2B20 colonizou o trato digestivo dos camundongos gnotobióticos e o número de células víaveis flutuou entre 10(9) e 10(10) ufc/g de fezes. Em ambos os grupos experimental e controle, S. Typhimurium se estabeleceu rapidamente numa faixa de 10(8) a 10(10) ufc/g de fezes e se manteve em níveis elevados até os animais morreram ou serem sacrificados. Em conclusão, o tratamento prévio de camundongos com L. acidophilus UFV-H2B20 protege os animais contra a infecção experimental com S. Typhimurium mas essa proteção não se deve à uma redução das populações patogênicas nos intestinos.