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Background: Histopathology is widely used for the diagnosis of inflammatory bowel disease in dogs. Variations in lesions and unavailability of uniform grading systems limit the usefulness of histologic examination. Hypothesis: CD3 cell numbers in chronic enteropathies of dogs correlate with clinical activity of the disease and with severity of histopathologic changes. Animals: Nineteen client-owned dogs examined because of chronic diarrhea, vomiting, or both. Methods: Samples of duodenal and colonic mucosa were collected endoscopically before and after treatment. Dogs that responded to a hypoallergenic diet were grouped as food-responsive diarrhea dogs (FRD, n = 10). Dogs with no clinical improvement after 10 days of treatment then received prednisolone (immunosuppressive doses) and were grouped as steroid-responsive diarrhea dogs (SRD, n = 9). Histopathologic assessment with a standardized grading system was performed retrospectively on the intestinal samples. Histologic score, total number of infiltrating cells, and CD3-positive cells were counted and compared with the clinical scoring. Results: No statistically significant difference was detected among histologic grading, total number of cells in the lamina propria, and T-cell numbers in biopsies before and after treatment in either group (FRD and SRD). Conclusions and Clinical Importance: Currently used histopathologic grading scores, total numbers of cells, and numbers of CD3-positive cells did not allow differentiation between FRD and SRD and did not correlate with clinical response to therapy. Based on these results, new grading scores assessing other criteria than total cell numbers and CD3-positive cells should be evaluated in the future. [ABSTRACT FROM AUTHOR]

Background: Inflammatory bowel disease (IBD) is thought to result from a dysregulated interaction between the host immune system and commensal microflora. Toll-like receptors (TLRs) recognize microbe-associated molecular patterns (MAMPs), but their role in enteropathies in dogs is unknown. Hypothesis: That there is a dysregulation of TLRs recognizing bacterial MAMPs in dogs with IBD. Animals: Sixteen healthy beagles and 12 dogs with steroid-treated (ST) and 23 dogs with food-responsive (FR) diarrhea. Methods: Prospective, observational study. mRNA expression of canine TLR2, 4, and 9 was evaluated by quantitative real-time RT-PCR in duodenal and colonic biopsies obtained before and after standard therapy. Samples from control dogs were taken at necropsy, with additional biopsies of stomach, jejunum, ileum, and mesenteric lymph node in 6 dogs. Results: There were significant differences ( P≤ .017) in expression of TLR2, 4, and 9 between the 6 sampled locations in healthy control dogs (lymph node > small intestine ≥ colon). Before therapy, ST expressed more mRNA than control dogs for all 3 receptors ( P < .05). There were no significant differences between pretreatment and posttreatment values, even though 32/35 dogs improved clinically. No associations were found when comparing receptor mRNA expression with either histology or clinical activity scores. Conclusions and Clinical Importance: Bacteria-responsive TLR2, 4, and 9 are upregulated in duodenal and colonic mucosa in IBD. This might lead to increased inflammation through interaction with the commensal flora. The absence of significant changes after therapy despite clinical improvement might point toward the existence of a genetic predisposition to IBD as described in human IBD. [ABSTRACT FROM AUTHOR]

Repair processes of the inflamed intestine are very important for dissolution of chronic enteropathies (CE). Therefore, we examined the mRNA abundance of growth hormone receptor (GHR), insulin-like growth factors (IGF)-1 and -2 in duodenal and colonic biopsies of dogs with CE such as food-responsive diarrhoea (FRD) and inflammatory bowel disease (IBD) before and after treatment as compared with each other and healthy dogs. A clinical score (Canine IBD Activity Index = CIBDAI) was applied to judge the severity of CE. Biopsies of duodenum and colon from client-owned dogs with CE were sampled before (FRD(bef), n = 5; IBD(bef), n = 5) and after treatment (FRD(aft), n = 5; IBD(aft), n = 5). Intestinal control samples were available from a homogenous control population (n = 15; C). Intestinal samples were homogenized, total RNA was extracted, reverse transcribed and analysed by real-time polymerase chain reaction to measure mRNA levels of GHR, IGF-1 and IGF-2. Results were normalized with glyceraldehyde phosphate dehydrogenase as housekeeping gene. The CIBDAI decreased during the treatment period in FRD and IBD (P < 0.01). In duodenum, GHR mRNA levels were higher in all groups than in C (P < 0.001). Duodenal IGF-1 mRNA levels in FRD(aft) and IBD(aft) tended to be higher than in C (P < 0.1). The IGF-2 mRNA abundance in FRD(aft) was higher than in C (P < 0.05) in duodenum. In colon, mRNA levels of IGF-1 in IBD(aft) were higher than in FRD(aft) (P < 0.05) and levels differed between IBD(aft) and C (P < 0.05). In conclusion, mRNA levels of GHR, IGF-1 and IGF-2 in the gastrointestinal tract were increased during CE when compared with gastrointestinally healthy dogs. The data suggest that GHR, IGF-1 and IGF-2 are involved in gastrointestinal repair processes.

There is evidence that probiotics have immune-modulating effects on intestinal inflammation during chronic enteropathies (CE). In an ex vivo culture system we investigated the influence of probiotics on mRNA and protein expression levels of cytokines in intestinal samples from dogs suffering from CE. Duodenal samples of client-owned dogs with CE (group CE; n = 12) were collected during diagnostic endoscopy. Additional duodenal samples of gastrointestinally healthy dogs (group C; n = 4) from an unrelated study were available. Based on histopathological analyses, no pathological changes or only mild to moderate eosinophilic and/or lymphoplasmacytic duodenitis were diagnosed. Tissue samples were cultured: (1) with cell culture medium alone (negative control), (2) with a probiotic cocktail (PC), constituted of three Lactobacilli spp. from healthy canine fecal isolates, (3) with the individual strains of PC, and (4) with a placebo powder. Viability of intestinal tissue and probiotic bacteria before and after culture was evaluated. The mRNA abundance of interleukin (IL)-10, IL-12p40, interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, and transforming growth factor (TGF)-beta1 was analyzed by real-time polymerase chain reaction (RT-PCR). Protein concentrations of IFN-gamma and IL-10 were measured in culture supernatant by ELISA. Results of RT-PCR were expressed as 2(-2DeltaCrossing Point) x 100 after normalization with beta-actin. There was a loss of about 1 log CFU/mL of probiotic bacteria during the incubation period. Viability of tissue was maintained as confirmed by non-significant release of lactate dehydrogenase. In C, addition of PC increased IL-10 mRNA levels (P < 0.1). In CE, PC increased mRNA and protein levels of IL-10 (P < 0.05). On the mRNA level, the ratio of TNFalpha-/IL-10, IFN-gamma/IL-10, and IL-12p40/IL-10 decreased after addition of PC (P < 0.05). The results demonstrate favorable effects of PC on regulatory cytokines relative to inflammatory cytokines that might contribute to reduction of intestinal inflammation.

The somatotropic axis regulates growth of the gastrointestinal tract (GIT). In addition, colostrum feeding and glucocorticoids affect maturation of the GIT around birth in mammals. We have measured mRNA levels of members of the somatotropic axis to test the hypothesis that colostrum intake and dexamethasone treatment affect respective gene expression in the GIT. Calves were fed either colostrum or an isoenergetic milk-based formula, and in each feeding group, half of the calves were treated with dexamethasone (DEXA; 30 microg/kg body weight per day). Individual parameters of the somatotropic axis differed (P < 0.05) among different GIT sections and formula feeding increased (P < 0.05) mRNA levels of individual parameters at various sites of the GIT. Effects of DEXA on the somatotropic axis in the GIT partly depended on different feeding. In colostrum-fed calves, DEXA decreased (P < 0.05) mRNA levels of IGF-I (esophagus, fundus, duodenum, and ileum), IGF-II (fundus), IGFBP-2 (fundus), IGFBP-3 (fundus), IGF1R (esophagus, ileum, and colon), IGF2R (fundus), GHR (fundus), and InsR (esophagus, fundus), but in formula-fed calves DEXA increased mRNA levels of IGF-I (esophagus, rumen, jejunum, and colon). Furthermore, DEXA increased (P < 0.05) mRNA levels of IGF-II (pylorus), IGFBP-3 (duodenum), IGF2R (pylorus), and GHR (ileum), but decreased mRNA levels of IGFBP-2 (ileum), and IGF1R (fundus). Whereas formula feeding had stimulating effects, effects of DEXA treatment on the gene expression of parameters of the somatotropic axis varied among GIT sites and partly depended on feeding.

The neonatal development of the gastrointestinal tract around parturition in precocious mammals is greatly affected by endocrine factors like glucocorticoids as well as by nutritional factors. We have studied the effects of glucocorticoids and colostrum supply on intestinal morphology, cell proliferation, digestive enzyme activities, and xylose absorption in neonatal calves to test the hypothesis that the intestinal development in neonatal calves is influenced by glucocorticoids, dependent on colostrum feeding. Calves designated GrFD(-) and GrFD(+) were fed a milk-based formula, whereas those designated GrCD(-) and GrCD(+) received colostrum. Dexamethasone (DEXA; 30 microg/kg/day) was injected at feeding times to calves of GrFD(+) and GrCD(+). On day 3, the D-xylose absorption was measured. The calves were euthanized on day 5 of life. Colostrum feeding increased villus sizes in jejunum and ileum, enhanced xylose absorption capacity, and increased peptidase activities in the ileum. DEXA treatment diminished sizes and cell proliferation rates of Peyer's patches in the ileum, yet increased proliferation of crypt cells in the ileum of formula-fed calves. DEXA reduced aminopeptidase N activities in the jejunum of formula-fed calves, but increased the peptidase activities mainly of colostrum-fed calves in the ileum. Thus, DEXA effects depended on intestinal segment and on different feeding, resulting in stimulation of crypt cell proliferation in the less mature ileum (of formula-fed calves) and in stimulation of peptidase activities in the more mature ileum (of colostrum-fed calves). We conclude that the effects of DEXA were related to the developmental stage of the neonatal intestine and promoted the intestinal development, depending on the developmental stage., (Copyright 2004 S. Karger AG, Basel)

The mammalian gastrointestinal tract is a complex consisting of the intestinal epithelial cells, the mucosal immune cells and the endogenous microflora. Since a couple of years, probiotics are used in animal production to improve performance. But there are only few results available on the immune modulating effects of pro- and prebiotics in human and veterinary medicine. Recent studies are promising that pro- and prebiotics can be used as an alternative to conventional therapy with immune suppressive drugs in patients with inflammatory bowel disease or dietary hypersensitivity. Yet further randomized and placebo-controlled studies are needed to clarify these mechanisms in vivo.

Concentrations of nonnutritional factors, such as insulin-like growth factor-I (IGF-I), in bovine colostrum are high and can modulate neonatal gastrointestinal tract development and function. In neonatal calves, we have investigated effects on intestinal epithelial cell morphology, proliferation, and absorption of feeding milk-born human IGF-I (hIGF-I) or a bovine colostrum extract. Calves were fed a milk-based formula containing amounts of nutrients comparable to colostrum for the first 3 d and a milk replacer from d 4 on. Formula and milk replacer contained only traces of nonnutritional factors. In experiment 1, supraphysiological amounts of hIGF-I (3.8 mg/L formula; secreted by transgenic rabbits with their milk) were added to the formula. Xylose appearance in blood (after feeding xylose on d 5) and intestinal traits (after euthanasia on d 8) did not differ between groups. In experiment 2, an extract of first-milked bovine colostrum that provided physiological amounts of IGF-I (0.50, 0.15, and 0.09 mg of IGF-I/L formula on d 1, 2, and 3, respectively, and 0.09 mg of IGF-I/L milk replacer on d 4) was added to formula or milk replacer. Plasma xylose concentration in the control group was transiently higher than in calves fed the colostrum extract. On d 5 (after euthanasia), villus circumferences and heights in small intestine, and epithelial cell proliferation rate in intestine were higher in calves fed the colostrum extract than in controls. In conclusion, orally administered hIGF-I from transgenic rabbits had no effect on the intestinal tract. However, feeding a bovine colostrum extract enhanced intestinal villus size, although it appeared to transiently decrease the absorptive capacity.