Avaliação da capacidade imunomodulatória de isolados de Lactobacillus bovinos (L38 E L36) em modelo experimental murino desafiado por Salmonella enterica subsp. enterica sorovar Typhimurium
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFMG |
Texto Completo: | http://hdl.handle.net/1843/BUBD-933JKG |
Resumo: | Developed countries have imposed severe restrictions on the use of antibiotics as growth promoters in cattle. In this scenario, probiotics stand out as potential substitutes for these traditional growth promoters. Probiotics are "live microorganisms which when administered in adequate amounts, confer health benefits to the host." Their mechanisms of action are: direct antagonism against pathogens, increased efficiency of intestinal barrier and immunomodulation. In addition, probiotcis have important protective effects against various diseases, such as salmonellosis which represents a major cause of economic losses in livestock production. There are several bacterial genera with probiotic effect, especially Lactobacillus. As most of theprobiotic preparations for animal nutrition uses human probiotic strains there are few knowlegment of the effect of potentially probiotic bacteria of bovine origin on the host health. Therefore, the objective of this study was to evaluate the ability of colonization and immunomodulatory effect of isolates L36 (L. acidophilus) and L38 (L. salivarius), obtained from feces of calves. To evaluate the ability of colonization and immunomodulatory effect, gnotobiotic mice were obtained by monoassociation of isolates L36 and L38 in germ-free animals and accompanied population levels in the faeces over ten days. The protective imunomodulatorory effect of the isolates were evaluated in conventional animals treated with L36 and L38 and challenged with Salmonella enterica subsp. enterica serovar Typhimurium. It was evaluated general health parameters (weight variation, hepatic and spleenic indexis), production of IgA in the intestinal mucosa, histological aspect of bowel and liver and cytokine profile (IL5, IL6, IL10, IL -12-p40, IL17a, Ifng, Tgfb1 and Tnfa) along the intestine portions by RT-qPCR. The isolates L36 and L38 were able to colonize the intestinal mucosa of germ-free animals, presented high population levels in feces (8.87 ± 0.38 log CFU / g of feces for L38 and 8.39 ± 0.83 log CFU / g of feces for L36). Furthermore, in gnotobiotic animals, L36 appears to stimulate a cellular response in the intestinal mucosa, type TH17, it leads to significant increases (p <0.05) of expression levels of IL6, IL17a, Tgfb1 and Tnfa. However, L38 produced a cytokine profile (increased IL5,IL10, IL12b, IL17a, Ifng and Tgfb1) in the intestine indicative of a balance between TH and Treg type responses. The isolates L36 and L38 showed up safe due to the absence of parameters that are indicative of local or systemic infection. Both isolates were unable to stimulate the production of IgA in the mucosa, both in gnotobiotic and conventional animals but they decreased IgA production in Salmonella-challenged animals. There was no protection afforded by isolates against Salmonella infection. However, in Salmonella-challenged animals, L38 led to high production of IL10 along all the intestine. Therefore, L36 and L38 showed up safe and produce different immunomodulatory profiles in the mucosal. |
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Avaliação da capacidade imunomodulatória de isolados de Lactobacillus bovinos (L38 E L36) em modelo experimental murino desafiado por Salmonella enterica subsp. enterica sorovar TyphimuriumSalmonellaProbióticosImunomodulaçãoRT-qPCRLactobacillusGenéticaDeveloped countries have imposed severe restrictions on the use of antibiotics as growth promoters in cattle. In this scenario, probiotics stand out as potential substitutes for these traditional growth promoters. Probiotics are "live microorganisms which when administered in adequate amounts, confer health benefits to the host." Their mechanisms of action are: direct antagonism against pathogens, increased efficiency of intestinal barrier and immunomodulation. In addition, probiotcis have important protective effects against various diseases, such as salmonellosis which represents a major cause of economic losses in livestock production. There are several bacterial genera with probiotic effect, especially Lactobacillus. As most of theprobiotic preparations for animal nutrition uses human probiotic strains there are few knowlegment of the effect of potentially probiotic bacteria of bovine origin on the host health. Therefore, the objective of this study was to evaluate the ability of colonization and immunomodulatory effect of isolates L36 (L. acidophilus) and L38 (L. salivarius), obtained from feces of calves. To evaluate the ability of colonization and immunomodulatory effect, gnotobiotic mice were obtained by monoassociation of isolates L36 and L38 in germ-free animals and accompanied population levels in the faeces over ten days. The protective imunomodulatorory effect of the isolates were evaluated in conventional animals treated with L36 and L38 and challenged with Salmonella enterica subsp. enterica serovar Typhimurium. It was evaluated general health parameters (weight variation, hepatic and spleenic indexis), production of IgA in the intestinal mucosa, histological aspect of bowel and liver and cytokine profile (IL5, IL6, IL10, IL -12-p40, IL17a, Ifng, Tgfb1 and Tnfa) along the intestine portions by RT-qPCR. The isolates L36 and L38 were able to colonize the intestinal mucosa of germ-free animals, presented high population levels in feces (8.87 ± 0.38 log CFU / g of feces for L38 and 8.39 ± 0.83 log CFU / g of feces for L36). Furthermore, in gnotobiotic animals, L36 appears to stimulate a cellular response in the intestinal mucosa, type TH17, it leads to significant increases (p <0.05) of expression levels of IL6, IL17a, Tgfb1 and Tnfa. However, L38 produced a cytokine profile (increased IL5,IL10, IL12b, IL17a, Ifng and Tgfb1) in the intestine indicative of a balance between TH and Treg type responses. The isolates L36 and L38 showed up safe due to the absence of parameters that are indicative of local or systemic infection. Both isolates were unable to stimulate the production of IgA in the mucosa, both in gnotobiotic and conventional animals but they decreased IgA production in Salmonella-challenged animals. There was no protection afforded by isolates against Salmonella infection. However, in Salmonella-challenged animals, L38 led to high production of IL10 along all the intestine. Therefore, L36 and L38 showed up safe and produce different immunomodulatory profiles in the mucosal.Países desenvolvidos têm imposto fortes restrições ao uso de antibióticos como promotores de crescimento na pecuária bovina. Neste cenário, os probióticos se destacam como potenciais substitutos para esses tradicionais promotores de crescimento. Probióticos são micro-organismos vivos que, quando são administrados em quantidades adequadas, conferem benefícios à saúde do hospedeiro. Seus mecanismos de ação são: antagonismo direto contra patógenos,aumento da eficiência de barreira intestinal e imunomodulação, tendo importantes efeitos protetores contra diferentes doenças, como a salmonelose que representa uma das principais causas de perdas econômicas na pecuária. Vários são os gêneros bacterianos com capacidade probiótica, com destaque para os lactobacilos. Como a maior parte das preparações probióticas para nutrição animal possuilinhagens probióticas humanas não se sabe o efeito de bactérias potencialmente probióticas de origem bovina sobre a saúde do hospedeiro. Portanto, o objetivo deste trabalho foi avaliar a capacidade de colonização e o efeito imunomodulador dos isolados L36 (L. acidophilus) e L38 (L. salivarius), obtidos de fezes de bezerros.Para avaliar a capacidade de colonização e efeito imunomodulador, foram obtidos camundongos gnotobióticos pela monoassociação dos isolados L36 ou L38 em animais germ-free e acompanhados os níveis populacionais nas fezes ao longo de dez dias. O efeito protetor e imunomodulador dos isolados foram também avaliados em animais convencionais tratados com L36 ou L38 e desafiados com Salmonellaenterica subsp. enterica sorovar Typhimurium. Foram avaliados parâmetros gerais de saúde (variação de peso, índice hepático e esplênico), produção de IgA na mucosa intestinal, aspectos da histologia geral do intestino e fígado e perfil de expressão das citocinas (IL5, IL6, IL10, IL12b, IL17a, Ifng, Tgfb1 e Tnfa) ao longo do intestino, por RT-qPCR. Os isolados L36 e L38 foram capazes de colonizar a mucosa intestinal de animais isentos de germes, apresentados altos níveis populacionais nas fezes (8,87±0,38 Log UFC/g de fezes para L38 e 8,39 ±0,83 Log UFC/g de fezes para L36). Além disso, em animais gnotobióticos, L36 parece estimular a resposta, na mucosa intestinal, do tipo TH17, pois leva a aumentos significativos (p<0,05) dos níveis de expressão de IL6, IL17a, Tgfb1 e Tnfa. Os isolados L36 e L38 apresentaram-se seguros, pois não causaram mudanças nosparâmetros avaliados que são indicativos de infecção local ou sistêmica. Ambos os isolados não foram capazes de estimular a produção de IgA na mucosa, tanto em animais gnotobióticos quanto convencionais, mas reduziram a produção de IgA em animais desafiados com Salmonella. Porém, L38 produziu um perfil de citocinas(aumento de IL5, IL10, IL12b, IL17a, Ifng e Tgfb1) no intestino indicativo de um balanço entre respostas TH e Treg. Não houve proteção conferida pelos isolados contra infecção por Salmonella. Entretanto, L38 em animais desafiados levou a produção elevada de IL10 ao longo de todo o intestino. Portanto, L36 e L38 se apresentaram seguros e produzem perfis imunomodulatórios diferentes na mucosa.Universidade Federal de Minas GeraisUFMGAlvaro Cantini NunesElisabeth NeumannAnderson MiyoshiAdriana Abalen Martins DiasAna Maria Caetano de FariaRaphael Steinberg da Silva2019-08-12T08:45:48Z2019-08-12T08:45:48Z2012-11-09info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/1843/BUBD-933JKGinfo:eu-repo/semantics/openAccessporreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMG2019-11-14T18:16:23Zoai:repositorio.ufmg.br:1843/BUBD-933JKGRepositório InstitucionalPUBhttps://repositorio.ufmg.br/oairepositorio@ufmg.bropendoar:2019-11-14T18:16:23Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false |
dc.title.none.fl_str_mv |
Avaliação da capacidade imunomodulatória de isolados de Lactobacillus bovinos (L38 E L36) em modelo experimental murino desafiado por Salmonella enterica subsp. enterica sorovar Typhimurium |
title |
Avaliação da capacidade imunomodulatória de isolados de Lactobacillus bovinos (L38 E L36) em modelo experimental murino desafiado por Salmonella enterica subsp. enterica sorovar Typhimurium |
spellingShingle |
Avaliação da capacidade imunomodulatória de isolados de Lactobacillus bovinos (L38 E L36) em modelo experimental murino desafiado por Salmonella enterica subsp. enterica sorovar Typhimurium Raphael Steinberg da Silva Salmonella Probióticos Imunomodulação RT-qPCR Lactobacillus Genética |
title_short |
Avaliação da capacidade imunomodulatória de isolados de Lactobacillus bovinos (L38 E L36) em modelo experimental murino desafiado por Salmonella enterica subsp. enterica sorovar Typhimurium |
title_full |
Avaliação da capacidade imunomodulatória de isolados de Lactobacillus bovinos (L38 E L36) em modelo experimental murino desafiado por Salmonella enterica subsp. enterica sorovar Typhimurium |
title_fullStr |
Avaliação da capacidade imunomodulatória de isolados de Lactobacillus bovinos (L38 E L36) em modelo experimental murino desafiado por Salmonella enterica subsp. enterica sorovar Typhimurium |
title_full_unstemmed |
Avaliação da capacidade imunomodulatória de isolados de Lactobacillus bovinos (L38 E L36) em modelo experimental murino desafiado por Salmonella enterica subsp. enterica sorovar Typhimurium |
title_sort |
Avaliação da capacidade imunomodulatória de isolados de Lactobacillus bovinos (L38 E L36) em modelo experimental murino desafiado por Salmonella enterica subsp. enterica sorovar Typhimurium |
author |
Raphael Steinberg da Silva |
author_facet |
Raphael Steinberg da Silva |
author_role |
author |
dc.contributor.none.fl_str_mv |
Alvaro Cantini Nunes Elisabeth Neumann Anderson Miyoshi Adriana Abalen Martins Dias Ana Maria Caetano de Faria |
dc.contributor.author.fl_str_mv |
Raphael Steinberg da Silva |
dc.subject.por.fl_str_mv |
Salmonella Probióticos Imunomodulação RT-qPCR Lactobacillus Genética |
topic |
Salmonella Probióticos Imunomodulação RT-qPCR Lactobacillus Genética |
description |
Developed countries have imposed severe restrictions on the use of antibiotics as growth promoters in cattle. In this scenario, probiotics stand out as potential substitutes for these traditional growth promoters. Probiotics are "live microorganisms which when administered in adequate amounts, confer health benefits to the host." Their mechanisms of action are: direct antagonism against pathogens, increased efficiency of intestinal barrier and immunomodulation. In addition, probiotcis have important protective effects against various diseases, such as salmonellosis which represents a major cause of economic losses in livestock production. There are several bacterial genera with probiotic effect, especially Lactobacillus. As most of theprobiotic preparations for animal nutrition uses human probiotic strains there are few knowlegment of the effect of potentially probiotic bacteria of bovine origin on the host health. Therefore, the objective of this study was to evaluate the ability of colonization and immunomodulatory effect of isolates L36 (L. acidophilus) and L38 (L. salivarius), obtained from feces of calves. To evaluate the ability of colonization and immunomodulatory effect, gnotobiotic mice were obtained by monoassociation of isolates L36 and L38 in germ-free animals and accompanied population levels in the faeces over ten days. The protective imunomodulatorory effect of the isolates were evaluated in conventional animals treated with L36 and L38 and challenged with Salmonella enterica subsp. enterica serovar Typhimurium. It was evaluated general health parameters (weight variation, hepatic and spleenic indexis), production of IgA in the intestinal mucosa, histological aspect of bowel and liver and cytokine profile (IL5, IL6, IL10, IL -12-p40, IL17a, Ifng, Tgfb1 and Tnfa) along the intestine portions by RT-qPCR. The isolates L36 and L38 were able to colonize the intestinal mucosa of germ-free animals, presented high population levels in feces (8.87 ± 0.38 log CFU / g of feces for L38 and 8.39 ± 0.83 log CFU / g of feces for L36). Furthermore, in gnotobiotic animals, L36 appears to stimulate a cellular response in the intestinal mucosa, type TH17, it leads to significant increases (p <0.05) of expression levels of IL6, IL17a, Tgfb1 and Tnfa. However, L38 produced a cytokine profile (increased IL5,IL10, IL12b, IL17a, Ifng and Tgfb1) in the intestine indicative of a balance between TH and Treg type responses. The isolates L36 and L38 showed up safe due to the absence of parameters that are indicative of local or systemic infection. Both isolates were unable to stimulate the production of IgA in the mucosa, both in gnotobiotic and conventional animals but they decreased IgA production in Salmonella-challenged animals. There was no protection afforded by isolates against Salmonella infection. However, in Salmonella-challenged animals, L38 led to high production of IL10 along all the intestine. Therefore, L36 and L38 showed up safe and produce different immunomodulatory profiles in the mucosal. |
publishDate |
2012 |
dc.date.none.fl_str_mv |
2012-11-09 2019-08-12T08:45:48Z 2019-08-12T08:45:48Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/1843/BUBD-933JKG |
url |
http://hdl.handle.net/1843/BUBD-933JKG |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Minas Gerais UFMG |
publisher.none.fl_str_mv |
Universidade Federal de Minas Gerais UFMG |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFMG instname:Universidade Federal de Minas Gerais (UFMG) instacron:UFMG |
instname_str |
Universidade Federal de Minas Gerais (UFMG) |
instacron_str |
UFMG |
institution |
UFMG |
reponame_str |
Repositório Institucional da UFMG |
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Repositório Institucional da UFMG |
repository.name.fl_str_mv |
Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG) |
repository.mail.fl_str_mv |
repositorio@ufmg.br |
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1816829901259407360 |