RESUMO
Different studies in animal rearing claim the probiotic potential of species of the genus Propionibacterium. The effects of strains of Propionibacterium acidipropionici isolated from poultry intestine on microbiota activity and intestinal mucosa development were investigated in the early stage of rearing chicks and the safety of the dose used was investigated. The strains P. acidipropionici LET105 and LET107, administered as monoculture to chicks from the 1st to 14th day of life in a daily dose of 106 cfu/ml administered in the drinking water resulted harmless. The animals arrived at the expected weight for age and no differences were observed with respect to the food intake and water consumption related to control without bacteria administration. The analysis of microbiota composition revealed the presence of propionibacteria at the middle and end of the trial only in treated groups. Normal development of lactic acid bacteria and bifidobacteria, and slow colonisation by Bacteroides at the 7th day of the study was observed in the same groups. Analysis of the organic acids concentrations in the caecal content of birds revealed higher lactic acid and lower butyric acid production. Lower short chain fatty acids total concentration than expected during treatment was related to a better development of the gut mucosa. Increase in length of villus-crypt units, goblet cells counts and neutral mucins production were evidenced. Higher mucus secretion produced by dietary supplementation with propionibacteria could provide increased protection against pathogens.
Assuntos
Galinhas/microbiologia , Microbioma Gastrointestinal , Probióticos/farmacologia , Propionibacterium , Ração Animal , Animais , Butiratos/metabolismo , Ceco/imunologia , Ceco/microbiologia , Galinhas/crescimento & desenvolvimento , Galinhas/fisiologia , Dieta/veterinária , Ácidos Graxos Voláteis/metabolismo , Feminino , Mucosa Intestinal/imunologia , Mucosa Intestinal/microbiologia , Intestinos/imunologia , Intestinos/microbiologia , Ácido Láctico/metabolismo , Distribuição Aleatória , Aumento de Peso/efeitos dos fármacosRESUMO
Prebiotics may increase intestinal Fe absorption in anaemic growing rats. The present study evaluated the effects of high-performance (HP) inulin and oligofructose on factors that regulate Fe absorption in anaemic rats during the growth phase. Male Wistar rats aged 21 d of age were fed AIN-93G ration without Fe for 2 weeks to induce Fe-deficiency anaemia. The rats were fed on day 35 a control diet, or a diet with 10 % HP inulin, or a diet with 10 % oligofructose, without Fe supplementation. The animals were euthanised after 2 weeks, and segments of the duodenum, caecum, colon and liver were removed. The expression levels of proteins in the intestinal segments were assessed using Western blotting. The levels of serum, urine and liver hepcidin and the concentrations of IL-10, IL-6 and TNF-α in the caecum, colon and liver were measured using the ELISA test. HP inulin increased the expression of the divalent metal transporter 1 protein in the caecum by 162 % (P= 0·04), and the expression of duodenal cytochrome b reductase in the colon by 136 % (P= 0·02). Oligofructose decreased the expression of the protein ferroportin in the duodenum (P= 0·02), the concentrations of IL-10 (P= 0·044), IL-6 (P= 0·036) and TNF-α (P= 0·004) in the caecum, as well as the level of urinary hepcidin (P< 0·001). These results indicate that prebiotics may interfere with the expression of various intestinal proteins and systemic factors involved in the regulation of intestinal Fe absorption in anaemic rats during the growth phase.
Assuntos
Anemia Ferropriva/dietoterapia , Proteínas de Transporte de Cátions/metabolismo , Grupo dos Citocromos b/metabolismo , Mucosa Intestinal/metabolismo , Prebióticos , Regulação para Cima , Anemia Ferropriva/imunologia , Anemia Ferropriva/metabolismo , Anemia Ferropriva/patologia , Animais , Proteínas de Transporte de Cátions/agonistas , Ceco/imunologia , Ceco/metabolismo , Ceco/patologia , Colo/enzimologia , Colo/imunologia , Colo/metabolismo , Grupo dos Citocromos b/química , Grupo dos Citocromos b/genética , Duodeno/imunologia , Duodeno/metabolismo , Duodeno/patologia , Hepcidinas/sangue , Hepcidinas/metabolismo , Hepcidinas/urina , Mediadores da Inflamação/metabolismo , Mucosa Intestinal/imunologia , Mucosa Intestinal/patologia , Inulina/efeitos adversos , Inulina/uso terapêutico , Fígado/imunologia , Fígado/metabolismo , Fígado/patologia , Masculino , Oligossacarídeos/efeitos adversos , Oligossacarídeos/uso terapêutico , Tamanho do Órgão , Prebióticos/efeitos adversos , Ratos Wistar , Aumento de PesoRESUMO
Sepsis is a systemic inflammatory response resulting from local infection due, at least in part, to impaired neutrophil migration. IL-12 and IL-18 play an important role in neutrophil migration. We have investigated the mechanism and relative role of IL-12 and IL-18 in polymicrobial sepsis induced by cecal ligation and puncture (CLP) in mice. Wild-type (WT) and IL-18(-/-) mice were resistant to sublethal CLP (SL-CLP) sepsis. In contrast, IL-12(-/-) mice were susceptible to SL-CLP sepsis with high bacteria load in peritoneal cavity and systemic inflammation (serum TNF-alpha and lung neutrophil infiltration). The magnitude of these events was similar to those observed in WT mice with lethal CLP sepsis. The inability of IL-12(-/-) mice to restrict the infection was not due to impairment of neutrophil migration, but correlated with decrease of phagocytosis, NO production, and microbicidal activities of their neutrophils, and with reduction of systemic IFN-gamma synthesis. Consistent with this observation, IFN-gamma(-/-) mice were as susceptible to SL-CLP as IL-12(-/-) mice. Moreover, addition of IFN-gamma to cultures of neutrophils from IL-12(-/-) mice restored their phagocytic, microbicidal activities and NO production. Mortality of IL-12(-/-) mice to SL-CLP was prevented by treatment with IFN-gamma. Thus we show that IL-12, but not IL-18, is critical to an efficient host defense in polymicrobial sepsis. IL-12 acts through induction of IFN-gamma and stimulation of phagocytic and microbicidal activities of neutrophils, rather than neutrophil migration per se. Our data therefore provide further insight into the defense mechanism against this critical area of infectious disease.
Assuntos
Ceco/imunologia , Ceco/metabolismo , Interleucina-12/metabolismo , Ativação de Neutrófilo/imunologia , Subunidades Proteicas/metabolismo , Punções , Sepse/imunologia , Sepse/metabolismo , Animais , Ceco/microbiologia , Ceco/cirurgia , Movimento Celular , Interferon gama/biossíntese , Interleucina-12/deficiência , Interleucina-12/genética , Subunidade p40 da Interleucina-12 , Interleucina-18/deficiência , Interleucina-18/genética , Interleucina-18/metabolismo , Ligadura , Camundongos , Camundongos Knockout , Neutrófilos/citologia , Neutrófilos/imunologia , Neutrófilos/metabolismo , Óxido Nítrico/biossíntese , Fagocitose , Subunidades Proteicas/deficiência , Subunidades Proteicas/genética , Sepse/microbiologia , Sepse/prevenção & controle , Taxa de SobrevidaRESUMO
Antigenic macromolecules present in food can induce inflammatory allergic reaction in sensitized persons. The aim of the present work is the development of an animal model to detect food antigens based on hypersensitivity reaction after food ingestion. New Zealand rabbits were divided in 5 groups. Group 1 (GI): control. G2: Ovalbumin (OVA) sensitized. G3: sensitized and orally challenged with OVA. G4: OVA sensitized and phosphate buffer solution challenged (PBS). G5: sensitized and challenged with OVA. Samples from cecum were stained with Alcian Blue pH < 1 for mast cells and with silver method for enteroendocrine cells (EEC). Other samples were immunostained with anti CD5 and CD25 monoclonal antibodies. Specific IgE levels were detected by PCA. Histopathology of G5 showed patchy edema, lymphangiectasia and eosinophilic infiltration. Results were expressed as cells per HPF (high power field); Mast cells in G1: 1.33; G2: 12.80 and G5: 10.20. Enteroendocrine cells in surface epithelium: G1: 1.6; G2: 6.0; G5: 4.2 and in deep epithelium: G1: 3.0; G2: 12.0 and G5: 7.3. Lymphocytes CD5+ in G1: 24.21: G2: 22.12 and G5: 23.97 and CD25+ in G1: 12.10: G2: 14.30 and G5: 21.68. Group 3 were similar to G1 and G4 to G2. We observed: mast cells increased in number probably due to OVA induced response. EEC showed an increase in sensitized animals because of higher expression of cytoplasmatic granules or differentiation from stem cells. Decrease in EEC number in challenged groups was likely to be based on vesicles release. Total T cells showed no significant differences among groups. CD 25+ cells were higher in sensitized and challenged animals. We concluded that rabbit model of sensitization and oral challenge is valid to study ingested food antigens and potential digestive pathologic reactions.
Assuntos
Antígenos/isolamento & purificação , Modelos Animais de Doenças , Hipersensibilidade Alimentar/imunologia , Animais , Anticorpos Monoclonais , Ceco/imunologia , Contagem de Células , Células Enteroendócrinas , Imunoglobulina E/análise , Mastócitos , Coelhos , Linfócitos TRESUMO
1. Poultry granulocytes are not clearly distinguished from each other with haematoxylin-eosin (HE) stain; thus, histochemical techniques must be used. Three experiments were carried out using 4-week-old Leghorn chickens. 2. Three, 80-chicken groups were orally infected with (1) 10(8) colony forming units (CFUs) Salmonella enteritidis, or (2) 10(4) Eimeria tenella oocysts, or (3) 10(8) CFUs S. enteritidis + 10(4) E. tenella oocysts. Ten chickens from each group were euthanased and caecum samples obtained. Caecum samples were fixed in 10% formalin (buffered, pH 7.4) at 4, 8, 12 h, 1, 3, 5, 7, and 14 d post-inoculation (PI). 3. Samples were stained using three different staining techniques: HE for the identification of heterophils and eosinophils, Ziehl-Neelsen for mast cells, and p-phenilenediamine dihydrochloride plus pyrocatechol (PPD + PC) for eosinophils. 4. Birds from Experiment 1 showed no changes in the numbers of granulocytes. Birds from Experiments 2 and 3 showed higher numbers of heterophils in caecal mucosa and submucosa separately, on d 5 and 7. In Experiment 3, a decrease was observed in submucosal mast cells on d 3. Chickens from Experiments 2 and 3 showed increased numbers of mucosal mast cells between d 7 and 14. 5. PPD + PC positively stained eosinophils, but not heterophils. 6. Numbers of heterophils and mast cells were increased during the acute inflammatory process caused by E. tenella. Therefore, mast cells could play a role as primary inflammatory cells. Eosinophils seem not to be part of the inflammatory process caused by E. tenella.
Assuntos
Ceco/imunologia , Galinhas , Coccidiose/veterinária , Granulócitos/imunologia , Enteropatias Parasitárias/veterinária , Mucosa Intestinal/imunologia , Doenças das Aves Domésticas/imunologia , Salmonelose Animal/imunologia , Animais , Basófilos/imunologia , Ceco/microbiologia , Ceco/parasitologia , Coccidiose/imunologia , Coccidiose/parasitologia , Eimeria tenella/crescimento & desenvolvimento , Eosinófilos/imunologia , Granulócitos/microbiologia , Granulócitos/parasitologia , Histocitoquímica/veterinária , Enteropatias Parasitárias/imunologia , Enteropatias Parasitárias/parasitologia , Mucosa Intestinal/microbiologia , Mucosa Intestinal/parasitologia , Masculino , Mastócitos/imunologia , Doenças das Aves Domésticas/microbiologia , Doenças das Aves Domésticas/parasitologia , Salmonelose Animal/microbiologia , Salmonella enteritidis/crescimento & desenvolvimentoRESUMO
We analyzed the local and systemic cellular immune response in mongolian gerbils inoculated with Entamoeba histolytica. Two groups were intracecally inoculated with E. histolytica trophozoites and two groups were used as controls. A inoculated groups and a control groups were sacrificed on days 10 and 30 post inoculation (p.i), the spleen and mesenteric lymph nodes (MLNs) lymphocytes (Ly) were isolated and incubated in vitro with 2 different amebic antigens. The proliferative Ly response of inoculated groups was greater than the Ly mitogenic response seen in control groups, at day 10 p.i. as well as day 30 p.i. (alpha = 0.05). Ly response of MLNs was greater in comparison to those of the spleen (alpha = 0.05). In other four groups, intradermal reactions with a antigen were used to demonstrated delayed hypersensitivity in gerbils after being inoculated with E. histolytica trophozoites. The percentage of volume increase of the plantar pad swelling were measured. Groups inoculated presented greater increases (alpha = 0.05) than groups controls. The evidence presented herein demonstrates that the presence of E. histolytica trophozoites in cecum induced a local and systemic cellular immune response.
Assuntos
Ceco/imunologia , Entamoeba histolytica/imunologia , Entamebíase/imunologia , Gerbillinae/imunologia , Animais , Antígenos de Protozoários/imunologia , Entamoeba histolytica/crescimento & desenvolvimento , Hipersensibilidade Tardia/imunologia , Imunidade Celular , Mucosa Intestinal/imunologia , Linfonodos/imunologia , Ativação Linfocitária , Masculino , Testes Cutâneos , Baço/imunologiaRESUMO
We have investigated the role of IgE in the local immunity of intestinal amebiasis, a parasitic infection known to induce specific antibody-forming cells (AFC) and IgA antibodies in rodents and humans. We found that intragastric immunization of rats with glutaraldehyde-fixed Entamoeba histolytica trophozoites significantly increased antiameba AFC in the Peyer's patches and spleen and that the lamina propria of the cecum from immunized animals was infiltrated by eosinophils armed with IgE antibodies. Morphometric analysis showed that IgE-containing cells and eosinophils were nearly three times more abundant in the cecum of immunized rats. Antigenic challenge with amebal lysates provoked an increase in the short-circuit current and in the transepithelial potential difference in Ussing-chambered cecum preparations from immunized rats. Although eosinophilia and the increase of IgE are common consequences of infection by parasitic worms, our results indicate that local immunity in intestinal amebiasis also involves IgE deposition, eosinophil infiltration, and type I hypersensitivity, which may explain some symptoms of amebic dysentery such as colic, abdominal tension, tenesmus, and bloody stools.
Assuntos
Antígenos de Protozoários/imunologia , Entamoeba histolytica/imunologia , Eosinofilia/imunologia , Mucosa Gástrica/imunologia , Hipersensibilidade Imediata/imunologia , Imunoglobulina E/fisiologia , Amebíase/etiologia , Amebíase/imunologia , Animais , Células Produtoras de Anticorpos/citologia , Ceco/imunologia , Relação Dose-Resposta Imunológica , Eletrofisiologia , Imunização , Infusões Parenterais , Mucosa Intestinal/imunologia , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
Caecal biopsy specimens from Jamaican children with the Trichuris dysentery syndrome (TDS) and age matched Jamaican controls were investigated by immunohistochemistry and by light microscopy. Biopsy specimens from all children (with TDS and controls) showed a mild to moderate increase in inflamatory cells. Except in the vicinity of the worm, where the epithlium was flattened, there was no other epithelial abnormality. Compared with controls, children with TDS had increased IgM lamina propria plasma cells and decreased intaepithelial T cells. There was also an increase in crypt epithelial cells proliferation. Lamina propia T cells (both activated and non-activated) were no more common in children with the Trichuris syndrome than controls. Epithelial cell HLA-DR and VLA-1 expression (which are increased in other colitides) were the same in both groups. Despite the presence of large worm burdens and chronic dysentery, therefore, only minor changes were seen in the caecal mucosa of children with TDS. (AU)