ponding copy number was calculated. The following reference strains were used as standards: Bifidobacterium longum subsp. longum CECT 4503, Bacteroides fragilis DSMZ 2451; Clostridium coccoides DSMZ 933; C. leptum DSMZ 935; Lactobacillus casei ATCC 393; and E. coli CECT 4558. Cytokine protein assay Jejunal tissue sections were kept in Krebs’s buffer supplemented with protease inhibitors until analysis. Tumour necrosis factor-a, and interleukine -10 were determined by ELISAs according to the manufacturer’s instructions. Prior to cytokine determination, tissue samples were homogenised in cell lysis buffer using a TissueRuptor. Then, samples were centrifuged to get clear supernatants for cytokine determination. The results of the ELISA assay are expressed as pico-grams per gram of tissue. Statistical analyses Statistical analyses were performed using SPSS v.15 software. For normally distributed data ANOVA and the Student t test were applied and, for nonnormally distributed data, the Mann-Whitney U test was used. Statistical significance was established at P,0.05 for all comparisons. Results Body weight and morphometric analyses of jejunal sections Significant differences in body weight were not detected among the different experimental animal groups. The BIX01294 site animals sensitised with interferon -c and fed gliadin were the only ones that presented signs of diarrhoea. The animal group that was diagnosed with diarrhoea had faecal spots around the anal area and the colon contents recovered after the Microbiological analyses Aliquots of different biologic samples were diluted in PBS and decimal dilutions were plated on MRS-C agar supplemented with mupirocin and B. longum CECT 7347 in an Enteropathy Animal Model sacrifice had watery consistency. Morphometric analyses of jejunal tissue sections revealed that animals fed with gliadin alone did not exhibit significant alterations compared to controls, except for decreased enterocyte height, but this alteration was restored by simultaneous administration of B. longum CECT 7347. In animals sensitised with interferon -c and fed gliadin, there was a significant decrease in villi width and enterocyte height and an increase in enterocyte numbers in the apical part of jejunal sections and also higher infiltration of cells in the lamina propria compared to the controls. However, these changes were not observed in animals only sensitized with IFN-c but not fed gliadin. B. longum CECT 7347 administration increased villi width and enterocyte height, partially restoring the alterations detected in animals sensitised with IFN-c and fed gliadin but did not decrease the cellular infiltration observed in the IFN/gliadin group. Feeding of B. longum CECT 7347 alone or together with gliadin increased villi length when compared to control animals. administration of both gliadin and B. PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22182644 longum CECT 7347 caused a significant increase in TNFa and IL-10 production, in comparison with the control and with the administration of either gliadin or B. longum CECT 7347 alone. In animals sensitised with IFN-c and fed gliadin, TNFa concentrations were significantly increased and, to a lesser extent, also those of IL-10 in comparison with control animals. This effect could be only attributed to the administration of gliadin in IFN-c sensitised animals because animals only sensitized with IFN-c but not fed with gliadin did not exhibit significant increases in cytokine production. The administration of B. longum CECT 7347 to the enteropath
