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Background Minimally invasive diagnostic imaging techniques to detect intestinal inflammation in dogs are lacking. Contrast-enhanced ultrasound (CEUS) and endoscopic narrow band imaging-like endoscopy (Storz Professional Image Enhancement System [SPIES]) might allow quantification of intestinal mucosal perfusion and microvessel density in chronic inflammatory enteropathy (CIE) of dogs. Hypothesis/objectives Markers of mucosal perfusion as determined by CEUS and SPIES endoscopy are potentially useful diagnostic markers to help characterize CIE and correlate with histological inflammation type and severity. Animals Thirty client-owned dogs diagnosed with CIE at a referral hospital were prospectively enrolled. Materials and methods Data from CEUS, SPIES, and white light (WL) endoscopy were correlated with World Small Animal Veterinary Association (WSAVA) endoscopy and histology scores and vessel density as determined by immunohistochemistry for von Willebrand factor (vWF). Automated linear modeling was used to determine predictors of endoscopic and histologic severity. Results Duodenal histology correlated with SPIES data (area percentage value, rho = 0.424, P = .04). White light endoscopy parameters correlated with SPIES parameters in the duodenum. Colonic histology correlated positively with CEUS, whereas colonic CEUS parameters correlated inversely with vWF expression. Several duodenal parameters combined predicted duodenal histology scores to a level of 73.4%. For the colon, 2 parameters contributed more than others to 73.1%. Conclusion and clinical importance Minimally invasive CEUS and SPIES appear feasible to assess intestinal perfusion in CIE. Use of SPIES endoscopy may be promising for assessing small intestinal inflammation, whereas CEUS could be used to assess colonic perfusion and inflammation. Both techniques need to be investigated further for their clinical utility.

Chronic diarrhoea is a frequent complaint in canine practice, and the diagnostic path is often characterized by numerous diagnostic tests and stepwise empirical treatments, often performed prior to gastrointestinal endoscopy/mucosal biopsies. These include dietary interventions (novel protein/hydrolysed protein diet), parasiticides and, still, in many cases, antibacterials. In order to decrease the risk of multiple potentially detrimental consequences for both the individual patient (antimicrobial resistance, long-term disruption of intestinal bacterial populations, potential worsening of gastrointestinal signs) and the general public, we advocate the use of antibacterials only after histopathologic evaluation of gastrointestinal biopsies, or at least after that any other therapeutic trial (e.g. diet/pre-probiotics, anti-inflammatory drugs) has been shown to be unsuccessful, in those cases in which endoscopy is not possible. They should be reserved, after appropriate dietary trials, for those canine chronic diarrhoeic patients with signs of true primary infection (signs of systemic inflammatory response syndrome or evidence of adherent-invasive bacteria) that justify antibacterial usage. In the present article we therefore discourage the empirical use of antibacterials when managing dogs with suspected chronic enteropathy.

Background Protein-losing enteropathy (PLE) because of chronic inflammatory enteropathy (CIE) in dogs is often treated with a combination of glucocorticoids and second-line immunosuppressant (SLI). This combined approach might not be necessary in all dogs. Hypothesis/objectives To describe diagnostic features and outcomes of dogs with PLE treated with glucocorticoids alone (group P) or with glucocorticoids and SLI (group S). Animals Thirty-one dogs with PLE. Material and methods Retrospective analysis of signalment data from diagnostic procedures, treatment, and outcome of dogs with CIE/PLE (from 2015 to 2017), using the hospital's digital case database. Dogs with hypoalbuminemia and CIE were included. Because of a stepwise treatment algorithm, dogs were allocated to group P or S. Time to serum albumin concentrations ≥20 g/L and survival data were collected. Dogs were additionally categorized by their albumin and cobalamin serum concentrations. Multivariate and univariate analysis as well as Pearson's correlation and Kaplan-Maier survival analysis were performed. Results Seventeen dogs were included in group P and 14 in group S. World Small Animal Veterinary Association score of the duodenum was different between groups (P = .05), but none of the other examined data. Median time until serum albumin reached >20 g/L was 13 days. Median survival time after start of treatment was 85 days (range, 13-463 days) in group P and 166 days (range, 8-390 days) in group S. Conclusion and clinical importance No routine diagnostic test was predictive of clinical response, treatment group, or outcome. Glucocorticoid treatment alone can be appropriate in dogs with PLE.

The current definition of probiotics was established by the Food and Agriculture Orga-nization of the United Nations and the World Health Organization in 2001 and 20021and amended by the International Scientific Association for Probiotics and Prebioticsin 2014,2and states that probiotics are “live microorganisms which when administered in adequate amounts confer a health benefit on the host”. Unfortunately, the label“ probiotic” is often misused as an umbrella term and applied to products that do not meet this strict definition.3In a further attempt to clarify what constitutes a probiotic, a newly created “probiotic framework”2separates live microbes used as pro-cessing aids or those in naturally fermented foods from those administered primarilyfor their health benefits (Fig. 1). It also dictates that, until any micro-organisms orgut commensals are isolated, characterized and their health effects are convincinglydemonstrated, they are not truly probiotics.

Hepatic encephalopathy is a frequent and debilitating complication of liver disorders. Lactulose is an established and reasonably effective treatment, yet with incompletely understood mechanisms of action. The aims of this study were to examine how the faecal microbiota composition changed before, during and after lactulose treatment in a large animal model. Healthy, privately owned dogs (n = 18) completed a prospective cohort study. Faecal samples were collected weekly, while the subjects were either on their usual diet (week 1), or a standardised diet (weeks 2–9), with added oral lactulose in weeks 6–7. DNA extraction and 16S rRNA gene sequencing were undertaken. Faecal samples from week 7 had a significantly lower microbiota richness/diversity, based on observed operational taxonomic units, Shannon/Chao1 indexes and Pielou’s Evenness. Beta diversity based on UniFrac distances was significantly different in week 7 compared to weeks 1, 5 and 9. At the phylum level, week 7 was associated with a significant increase of Firmicutes and Actinobacteria, and a decrease of Bacteroidetes and Fusobacteria, when compared to weeks 5 and 9. In summary, we have shown that lactulose induces a reversible qualitative and quantitative change of the faecal microbiota, which may explain its clinical efficacy in the management of hepatic encephalopathy.

BackgroundIdiopathic esophageal dysmotility (ED) is increasingly recognized in young dogs of brachycephalic breeds. Few studies have objectively associated specific videofluoroscopic swallowing study (VFSS) features with brachycephaly, leading to under‐recognition of ED in brachycephalic breeds.Hypothesis/objectivesTo describe and compare VFSS in brachycephalic dogs versus non‐brachycephalic dogs presented for dysphagia or regurgitation, and to investigate associations between these imaging findings and patient signalment.MethodsRetrospective analysis of VFSS of dogs presented for dysphagia or regurgitation (not megaesophagus) from 2006 to 2017. Cases were divided into brachycephalic and mesaticephalic breeds. The VFSS were reviewed using a standardized protocol by 2 examiners. Esophageal motility was assessed using specific criteria, and particular imaging features were noted and graded. Fisher's exact test was used to determine associations among signalment (including brachycephaly), final diagnosis, outcomes, and ED features.ResultsThirty‐six dogs were included (n = 10 normal, n = 26 presumed ED). Twenty dogs (77%) with presumed ED were brachycephalic with a median age of 1 year (range, 0.2‐10.5 years). Most common were prolonged esophageal transit time (ETT; n = 21/26), decreased propagation of secondary peristaltic waves (n = 20/26), and gastroesophageal reflux (GER; n = 18/28). Eight dogs (all brachycephalic) had hiatal herniation (HH). Morphological esophageal variations were only observed in brachycephalic dogs. Brachycephaly was significantly associated with ED (P = .005), prolonged ETT (P = .41), GER (P = .02), and HH (P = .03). Conclusions and Clinical ImportanceThe majority of dogs with presumed ED was young and brachycephalic and had specific abnormalities that were less frequent in mesaticephalic dogs with regurgitation or dysphagia.

Background: Dogs with hypoadrenocorticism (HA) frequently show gastrointestinal signs of gastrointestinal disease (SGD). The prevalence of dogs presented for chronic SGD with HA is unknown. Objectives: The aims of this study were to determine the prevalence of HA in dogs with chronic SGD and to identify clinical and laboratory variables for HA in this population. Animals: 151 dogs with chronic SGD Methods: In this multi-centered prevalence study a standardized work-up was performed in prospectively enrolled dogs. Basal serum cortisol concentration was measured in every dog with SGD >3 weeks, followed by an ACTH stimulation test (ACTHST) if basal serum cortisol concentration was

Background Canine chronic enteropathies (CE) are believed to be caused by an aberrant immune response towards the intestinal microbiome. Administration of probiotics can alleviate colitis in people. In vitro effects of the probiotic Enterococcus faecium NCIMB 10415 E1707 (EF) previously have been evaluated using canine cells (e.g., whole blood, intestinal biopsies), but data on in vivo efficacy are lacking. Hypothesis/Objectives Administration of EF to dogs with food-responsive CE will improve clinical outcome and decrease the intestinal inflammatory profile. Animals Dogs diagnosed with CE were prospectively recruited to receive a hydrolyzed elimination diet plus either a synbiotic product containing EF or placebo for 6 weeks. Both veterinary staff and owners were blinded to the treatment. Methods Clinical severity index (CCECAI), clinicopathological data and gene expression using intestinal biopsies (TLR2/4/5/9, IL-17A, IL-22, IL-23p19, RORC, IL-2, IL-12p35, TNFα, IL-4, IFNy, IL-10, TGFβ, IL-1β, IL-18, NLRP3, casp-1, TFF1, TFF3 and PPARy) before and after 6 weeks of treatment were analyzed using linear mixed modeling. Results Of the 45 cases recruited, 12 finished the clinical trial. Seven received the synbiotic and 5 the placebo product. There was no difference between groups or treatments regarding clinical efficacy, histology scores or expression of any of the investigated genes. Conclusions and clinical importance Standard dietary treatment induced rapid clinical response in all cases. Because the study was underpowered, it was not possible to determine whether or not EF had an additional effect within the time period of 6 weeks.

The composition of the microbiome plays a significant role in the pathogenesis of inflammatory bowel disease (IBD) in humans and chronic enteropathies (CE) in dogs. The administration of probiotic micro-organisms is one way of modulating the microbiome, but experiments elucidating mechanisms of action of probiotics in the intestine of healthy and CE dogs are lacking. The aim of our study was to investigate the effects of different Toll-like receptor (TLR) ligands and Enterococcus faecium (EF) on ex vivo cultured duodenal samples and whole blood (WB) from dogs with food-responsive chronic enteropathy (FRE) when compared to healthy dogs. Biopsy stimulation was performed in 17 FRE and 11 healthy dogs; WB stimulation was performed in 16 FRE and 16 healthy dogs. Expression of TLR2, 4, 5 and 9, IL-17A, IL-22, IFNy, TNFα, IL-4, IL-10, TGFβ and PPARy was determined in biopsies by quantitative polymerase chain reaction (PCR). In addition, production of TNFα, IL-10, IFNy and IL-17A protein in WB and biopsy supernatants was assessed by ELISA. Treatment with individual TLR ligands or EF induced a variety of changes in the expression of different TLRs and cytokines, but not necessarily a consistent change with a single stimulating agent. Even though cytokine protein could not be detected in supernatants from ex vivo stimulated biopsies, we found TNFα protein responses in blood to be opposite of the transcriptional responses seen in the biopsies. Stimulation of canine duodenal biopsies with TLR ligands can potentially induce anti-inflammatory gene expression, especially in healthy tissue, whereas the effects of EF were limited.

Canine idiopathic inflammatory bowel disease (IBD) is believed to result from complex interplay between genetic, microbial, and immunologic factors. Abnormal cell death by apoptosis may result in the persistence of activated intestinal T cells that contribute to mucosal inflammation and clinical severity. To test this hypothesis, we investigated the mucosal expression of pro- and anti-apoptotic proteins in different intestinal compartments and their association with inflammatory indices in dogs with IBD. Apoptosis of lamina propria (LP) T cells in duodenal, ileal, and colonic tissues in control and IBD dogs was analyzed by caspase 3/Bcl-2 immunohistochemistry and TUNEL assays. Densities and distributions of LP caspase 3 and Bcl-2 cells were correlated to histopathologic lesions and the clinical activity index (CIBDAI). Compared to control tissues, IBD dogs had significantly (P0.01) fewer caspase 3 cells in colonic mucosa. Double immunostaining identified the majority of apoptotic cells as TUNEL(+)/caspase 3(+). Within intestinal mucosa of IBD dogs, there were significantly greater numbers of Bcl-2 cells at the apical and basilar villus in the duodenum as compared to the colon and to the apical and basilar villus in the ileum (P0.001 for all comparisons). There were significantly greater numbers of Bcl-2 cells at the apical and basilar villus of the duodenum but significantly fewer numbers of Bcl-2 cells at the apical villus of the ileum in IBD dogs compared with controls (P0.001, P0.001, and P0.02, respectively). There was a significant association between the number of Bcl-2 cells in the duodenum of IBD dogs and the CIBDAI (P0.001 each for mild, moderate and severe clinical IBD). In conclusion, apoptosis of T lymphocytes varies within intestinal compartments of dogs with IBD. Mucosal imbalance of Bcl-2/caspase 3 expression favors T cell resistance to apoptosis which may contribute to T cell accumulation and chronic intestinal inflammation, similar to human IBD.

SummaryObjective: Delayed gastric emptying is a well-recognised phenomenon in a number of canine disease conditions. Only a limited number of drugs have been reported to have some gastrokinetic effect in the dog. The aim of this study was to investigate prokinetic effects of maropitant. Material and methods: In a cross-over study 24 healthy adult Beagle dogs were randomised to receive either maropitant (2 mg/kg q24 h PO), cisapride (1 mg/kg q12 h PO) or placebo (vitamin- B12, 10 μg/dog q24 h PO) for 7 days with a 7-day washout period between treatments. Gastric emptying was measured simultaneously via 99mTechnetium radioscintigraphy and 13C-sodium acetate breath testing for 6 hours post-feeding. The decrease in radioactive counts in the stomach and the increase in 13CO2 concentration in exhaled breath (measured via gas chromatography) were plotted against time. The area under the curve was determined for each test and the time to 25%, 50% and 75% gastric emptying was calculated for each method. Friedman test was used to compare gastric emptying times. Results: With both methods, no difference for gastric emptying times was observed for any treatment. Conclusion and clinical relevance: Neither maropitant nor cisapride were shown to have an effect on gastric emptying in healthy beagles using radioscintigraphy or breath test when compared to placebo. Consequently, neither drug can be recommended as a gastric prokinetic in dogs.

Inflammasomes coordinate the maturation of IL-1β and IL-18 in response to danger signals. They are vital for maintenance of intestinal homeostasis and have been linked to chronic intestinal inflammation in humans. Probiotics have been advocated as treatment in intestinal inflammation. So far, no study has investigated the role of the inflammasome in canine chronic enteropathy (CE). In this study the intestinal expression of inflammasome components was assessed in CE dogs compared to controls, when treated with probiotic Enterococcus faecium (EF) ex-vivo and in-vivo. RNA extraction from endoscopic biopsies and reverse-transcriptase quantitative PCR was performed for NLRP3, casp-1, IL-1β and IL-18. Immunohistochemistry was performed to investigate protein expression in tissues. Gene expression of casp-1 and NLRP3 was lower in CE samples than controls. Ex-vivo treatment with EF reduced NLRP3 expression in control samples. Treatment of CE dogs with EF alongside dietary intervention had no effect on gene expression. In contrast, IL-1β protein expression in CE decreased with dietary treatment (but not with probiotics). The results of this study suggest that the inflammasome or its components may be partially involved in the inflammatory process seen in CE, but distinct from intestinal inflammation in humans.

Objective—To directly compare solid-phase gastric emptying times assessed by means of a [13C]sodium acetate breath test ([13C]-SABT) and technetium Tc 99m albumin colloid radioscintigraphy (99mTc-ACR) in healthy cats. Animals—12 healthy cats. Procedure—After ingestion of a test meal containing 50 mg of [13C]sodium acetate and 250 MBq of 99mTc-albumin colloid, each cat underwent simultaneous [13C]-SABT and 99mTc-ACR on 2 consecutive days. Breath samples and scintigrams were acquired at 30, 60, 90, 120, 150, 180, 210, 240, 300, 360, 480, and 600 minutes after meal ingestion. Quartiles of gastric emptying (25%, 50%, and 75%) were calculated for breath test analysis by use of the area under the curve of the 13C:12C ratio. Quartiles of gastric emptying times were extrapolated from the scintigraphic findings by beans of nonlinear curve regression analysis. Results—Mean ± SD gastric half-emptying (50%) times obtained with [13C]-SABT and 99mTc-ACR, were 239 ± 28 minutes and 276 ± 59 minutes, respectively. A 2-way repeated-measures ANOVA revealed that mean gastric emptying times determined with [13C]-SABT and 99mTc-ACR differed significantly. For the stages of gastric emptying, Pearson correlation between the 2 methods was good at 25% (r = 0.655) and weak at 50% (r = 0.588) and 75% (r = 0.566)of gastric emptying. Conclusions and Clinical Relevance—Results indicated that the [13C]-SABT can be a valid alternative to 99mTc-ACR in healthy cats; it was easy to perform, was tolerated well by the cats, and had acceptable correlation to scintigraphic findings at gastric emptying of 25%, 50% and 75%. Studies in cats with delayed gastric emptying will be needed to verify the validity of the [13C]-SABT.

Trefoil factors (TFF) are small peptides produced by goblet cells, which are crucial for epithelial restitution. In humans with inflammatory bowel disease (IBD), TFF expression is up-regulated as part of an unspecific repair mechanism. The goal of this study was to assess TFF gene expression in the gastrointestinal tract from dogs with IBD compared to healthy controls. Preliminary assessment by PCR revealed TFF1 and 3 expression in the small and large intestine, whereas TFF2 was amplified only in the stomach. Subsequent RT-qPCR (with relative quantification against 3 reference genes) on endoscopic duodenal (IBD n = 22, healthy controls n = 18) and colonic (IBD n = 12, controls n = 11) biopsies revealed that TFF1 expression was significantly up-regulated in the duodenum from IBD dogs (Mann–Whitney p = 0.001), whereas TFF3 expression was significantly lower in IBD colon compared to controls (t-test p = 0.018). This study demonstrates evidence for dysregulation of TFF gene expression in canine IBD. Up-regulation of TFF1 could signify ectopic expression as a compensatory repair-mechanism, whereas down-regulation of TFF3 could contribute to defective epithelial barrier function, respectively. Whether this is a cause or consequence of IBD could not be established.

The assessment of in vitro responses of blood-derived cells has traditionally been performed with peripheral blood mononuclear cells (PBMCs). However, stimulation of whole blood (WB) has advantages: ease of experimental setup, avoidance of blood cell manipulation and lower assay cost and time. WB stimulation is widely used in human research, but only infrequently in small animals. The aim of this study was to compare the response generated in canine WB and PBMCs with Toll-like receptor ligands and probiotic bacteria using TNFα as measured endpoint. WB and PBMCs were derived from a total of 15 healthy dogs. Stimulations were performed with LPS (1 ng ml−1), Pam3CSK4 (100 ng ml−1), flagellin (1 μg ml−1) and Enterococcus faecium (EF; 1 × 107 cfu ml−1). In 4 of the dogs, PBMC numbers were matched to the numbers of PBMCs found in WB. TNFα was detected in supernatants via ELISA. TNFα production from WB was generally higher than from PBMCs (repeated measures ANOVA p < 0.0128). PBMCs produced TNFα inconsistently for all stimulants apart from EF. There was no correlation between results of WB or PBMC stimulation, similar to studies that found that humanWB cytokine production correlates with stimulating monocytes, but not PBMCs. In conclusion, WB stimulation should be considered a valid alternative to PBMC stimulation in the canine system.

Summary Objective: The aim of the study was to assess solid phase gastric emptying via non-invasive 13C-sodium acetate breath test in large breed dogs with or without gastric dilatation-volvulus (GDV). Material and methods: Dogs were recruited into one of the following groups: group 1 = healthy large breed dogs with no history of GDV, group 2 = dogs that underwent elective abdominal surgery for reasons unrelated to the gastrointestinal tract, and group 3 = dogs that underwent laparotomy and gastropexy to correct GDV. The dogs were fed a test meal containing 100 mg 13C-sodium acetate (for group 2 and 3, this was < 48 hours post-operatively). Breath samples were obtained at baseline and every 30 minutes for 3 hours, then every hour for a total of 7 hours. 12CO2/13CO2 ratio was measured for each breath sample via non-dispersive infrared spectroscopy and 25%, 50% and 75% gastric emptying times were calculated and compared between groups. Results: Gastric emptying times were significantly prolonged in dogs undergoing surgery (group 2) compared to group 1 and 3. Also, gastric emptying times of dogs with GDV were significantly prolonged compared to controls, but not to the same extent as dogs in group 2. Conclusion and clinical significance: There was a significant effect of abdominal surgery on gastric emptying times. Surprisingly, dogs after GDV surgery and gastropexy had shorter gastric emptying times than dogs undergoing laparotomy for reasons other than GDV, but still prolonged compared to healthy controls. The reason for these differences requires further study.

Inflammatory bowel disease (IBD) is a common cause of chronic diarrhoea in dogs. In people, specific cytokine patterns attributed to T cell subsets, especially T helper cell [Th]1, Th17 and regulatory T(reg) cells have emerged in IBD. In contrast, no specific involvement of a distinct T cell subset has been described so far in canine IBD. Thus, the aim of the present study was to assess gene expression of signature cytokines in duodenal tissues from 18 German shepherd dogs with IBD (group 1), 33 dogs of other breeds with IBD (group 2) and 15 control dogs (group 3). Relative quantification of IL-17A, IL-22, IL-10, IFNy and TGFβ was performed. Expression of IL-17A was significantly lower in groups 1 and 2 compared to group 3 (p = 0.014), but no difference in the expression of IL-22 (p = 0.839), IFNγ (p = 0.359), IL-10 (p = 0.085) or TGFβ (p = 0.551) across groups was detected. Thus, no clear evidence for the involvement of Th-17 signature cytokines in canine IBD at the mRNA level could be shown. The contribution of specific T cell subsets to the pathogenesis of canine IBD warrants further investigation.

CD11c serves as a marker for human and murine dendritic cells (DCs) and cells expressing this marker have been shown to have similar morphological and functional characteristics in the canine immune system. The aim of this study was to quantify CD11c(+) cells in the duodenum, ileum and colon of healthy dogs and dogs with inflammatory bowel disease (IBD). Endoscopic biopsies from the duodenum (n=12 cases), ileum (n=8 cases) and colon (n=12 cases) were obtained from dogs diagnosed with IBD. Intestinal tissue from 10 healthy beagle dogs was used as control. Immunofluorescence microscopy was carried out using an anti-canine CD11c monoclonal antibody. Labelled cells were recorded as cells per 120,000 μm(2). The canine chronic enteropathy clinical activity index (CCECAI) was calculated for all dogs with IBD. In addition, sections from all dogs with IBD were evaluated according to the guidelines of the World Small Animal Veterinary Association Gastrointestinal Standardization Group. The number of CD11c(+) cells in the duodenum, ileum and colon of dogs with IBD was significantly reduced compared with controls (P

Different antibody-based tests for rapid detection of Canine parvovirus antigens in feces are commercially available, allowing quick diagnosis in a clinical setting. However, the diagnostic accuracy of these tests compared with standard methods has not been evaluated so far. In the current study, 3 commercial tests were compared with immune-electron microscopy (IEM) and polymerase chain reaction (PCR). Dogs were divided into 3 groups: group A, samples from dogs with acute hemorrhagic diarrhea ( n = 50); group B, dogs with chronic diarrhea ( n = 10); and group C, dogs with no evidence of gastrointestinal disease ( n = 40). Specificity of all 3 commercial tests versus PCR and IEM was good to excellent (92.2–100%). Sensitivity, in contrast, was poor: 15.8–26.3% versus PCR and 50–60% versus IEM. In group A, 10 dogs were positive by IEM and 24 dogs were positive by PCR. Positive PCR results were also obtained from animals in control groups (group B, 1 dog; group C, 5 dogs). No dog in group B or C was positive by IEM. In conclusion, the rapid tests are useful to diagnose canine parvoviral enteritis, but they do not rule out parvovirus infection in an animal with typical clinical signs. In addition, a small percentage of healthy dogs and dogs with chronic diarrhea showed positive PCR results; this may be due to asymptomatic/persistent infection or intestinal passage of virus. The significance of this finding remains unclear.