Isolation of Malassezia yeasts from dogs with gastrointestinal disease undergoing duodenal endoscopy.

OBJECTIVES: To culture Malassezia and other fungi from the duodenum of dogs with gastrointestinal signs undergoing routine endoscopic examination. MATERIALS AND METHODS: Quantitative microbial culture was performed on duodenal juice aspirated from dogs with suspected enteropathy during routine upper gastrointestinal endoscopy; samples were cultured on Sabouraud's dextrose agar (30, 32 and 37°C) and modified Dixon agar (32°C) for 14 days. Isolates were identified phenotypically and by matrix-assisted laser desorption ionisation-time of flight, and internal transcribed spacer sequencing. Yeast presence was also evaluated by cytological and histopathological examination of smears and biopsy specimens. RESULTS: Forty-five dogs were recruited with chronic inflammatory enteropathy (n=38), granulomatous colitis (n=2), gastric adenocarcinoma (n=2), duodenal small cell lymphoma (n=1) and idiopathic severe gastrointestinal haemorrhage (n=2). Fungi were cultured from 14 dogs: Malassezia pachydermatis was isolated from eight [chronic inflammatory enteropathy (n=7) (along with Candida albicans n=1); granulomatous colitis (n=1)] and Malassezia sympodialis from another (gastric adenocarcinoma). Five dogs with chronic inflammatory enteropathy yielded other yeasts (C. albicans, Candida glabrata, Kazachstania slooffiae, Kazachstania telluris, Pichia kudriavzevii [syn. C. krusei]). Yeasts were never observed in histopathological specimens. Fluorescent microscopical examination of cytological specimens showed yeast in only one case, from which K. slooffiae was subsequently isolated. CLINICAL SIGNIFICANCE: Based on a literature search, this is the first report of isolation of M. pachydermatis, M. sympodialis, K. slooffiae and K. telluris from the canine duodenum. Further studies are needed to determine whether these are resident or transient fungi in the canine duodenum and whether their presence has a pathogenic effect on the host.

Outcome of chronic inflammatory enteropathy in cats: 65 cases (2011-2021).

OBJECTIVES: Feline chronic inflammatory enteropathy is an idiopathic disease with limited information on variables that might affect treatment outcome and survival. The aim of this study was to determine if clinicopathological variables were associated with death due to gastrointestinal disease in cats with chronic inflammatory enteropathy. MATERIALS AND METHODS: Three medical records databases were retrospectively searched for cats diagnosed with chronic inflammatory enteropathy at the Royal Veterinary College between June 2008 and November 2021. Intestinal biopsy specimens of eligible cases were re-reviewed by one board-certified veterinary pathologist. Outcome information was obtained by contact with the referring veterinary surgeon. Two univariable binary logistic regression models and a Fisher's exact test were performed to assess the association between the outcome of death due to gastrointestinal disease or its short-term survival (≤ versus >1 year) with clinicopathological variables and the attainment of clinical remission. RESULTS: Sixty-five cats diagnosed with chronic inflammatory enteropathy between September 2011 and August 2021 were included in the study with follow-up information available for 54 cats (83%). Of these 54 cats, 20 (37%) were euthanised due to gastrointestinal disease (median 129.5 days; range 8 to 2970 days). Twenty-five (46%) cats were alive and in clinical remission (median 916 days; range 78 to 2113 days) with 16 (64%) diagnosed with food-responsive enteropathy. Attaining clinical remission reduced the likelihood of subsequent death due to gastrointestinal disease. CLINICAL SIGNIFICANCE: Measured physical and laboratory variables at the time of histopathological diagnosis of chronic inflammatory enteropathy were not predictors of death. Alternative diagnostic measures are required to definitively investigate outcome and survival in cats with chronic inflammatory enteropathy.

The use of hydrolysed diets for vomiting and/or diarrhoea in cats in primary veterinary practice.

OBJECTIVE: To describe responses of cats prescribed a hydrolysed diet with or without concurrent medication for chronic vomiting and/or diarrhoea of undetermined aetiology. MATERIALS AND METHODS: Anonymised records of 512,213 cats under UK veterinary care in 2016 from the VetCompass database were searched using relevant terms for hydrolysed diets. The records of 5000 (90%) of 5569 cats with evidence of receiving a hydrolysed diet were randomly reviewed for gastrointestinal indication, prior and concurrent medication and response after hydrolysed dietary intervention. A poor response was defined as evidence of receiving antibiotic or glucocorticoid treatment for vomiting/diarrhoea at visits after the onset of the diet or death from gastrointestinal signs for at least 6 months follow-up. RESULTS: Of 977 cats prescribed a hydrolysed diet for chronic vomiting/diarrhoea, 697 (71%) were first prescribed the diet without concurrent antibiotics or glucocorticoids while 280 (29%) first received the diet with these medications. Thirty-four per cent of cats in the former group and 61% in the latter had a poor response. Cats older than 6 years and cats prescribed antibiotic and/or glucocorticoid for vomiting/diarrhoea before and concurrently with the diet had higher odds of poor response. CLINICAL SIGNIFICANCE: Although variations in our observations may reflect severity of signs or prescribing habits of primary-care veterinary surgeons, our study suggests there is merit in trialling a hydrolysed diet first as a sole therapy in cats with chronic vomiting/diarrhoea when diagnostic investigations do not reveal a cause, before resorting to antibiotic and/or glucocorticoid therapy for cases that respond poorly.

Serological markers of gluten sensitivity in Border terriers with gall bladder mucocoeles.

OBJECTIVES: To evaluate serological markers of gluten sensitivity in conjunction with cholecystokinin measurement in Border terriers with gall bladder mucocoeles. MATERIALS AND METHODS: Medical records from two referral hospitals were obtained between 2011 and 2019 to identify Border terriers with gall bladder mucocoeles, non-Border terriers with gall bladder mucocoeles and control Border terriers with non-biliary diseases. Enzyme-linked immunosorbent assays were performed on stored fasted serum samples for anti-gliadin IgG, anti-canine transglutaminase-2-IgA autoantibodies and cholecystokinin. Statistical analysis was performed using the Kruskall-Wallis test to identify differences between the groups. RESULTS: Fifteen Border terriers with gall bladder mucocoeles, 17 non-Border terriers with gall bladder mucocoeles and 14 control Border terriers with non-biliary diseases were recruited. Median transglutaminase-2-IgA autoantibodies in Border terriers with gall bladder mucocoeles was 0.73 (range: 0.18 to 1.67), which was significantly greater than in control Border terriers at 0.41 (0.07 to 1.14). Median cholecystokinin concentration in Border terriers with gall bladder mucocoeles was 13 pg/mL (6 to 45 pg/mL), which was significantly lower than in control Border terriers at 103 pg/mL (9 to 397 pg/mL). There was no difference in the anti-gliadin IgG between these groups. There was no difference observed in the non-Border terriers with gall bladder mucocoeles with either of the other groups. CLINICAL SIGNIFICANCE: Reduced cholecystokinin and increased transglutaminase-2-IgA autoantibodies was detected in Border terriers with gall bladder mucocoeles; which is in part homologous to gall bladder disease identified in human coeliac disease. The results suggest an immunological disease with impaired cholecystokinin release may be affecting gall bladder motility and possibly contributing to mucocoele formation in Border terriers.

Gall bladder mucoceles in Border terriers.

BACKGROUND: Gall bladder mucoceles (GBM) are a leading cause of biliary disease in dogs with several breeds, including the Shetland Sheepdog, American Cocker Spaniel, Chihuahua, Pomeranian, and Miniature Schnauzer apparently predisposed. OBJECTIVE: To determine risk factors, clinical features, and response to treatment of GBM in Border terriers (BT). ANIMALS: Medical records of 99 dogs (including 51 BT) with an ultrasonographic (±histopathologic) diagnosis of GBM from three referral centers in the United Kingdom were collected. A control group of 87 similar-aged BT with no ultrasonographic evidence of gall bladder disease was selected for comparison. METHOD: Retrospective case-control study. Odds ratios were calculated to establish breed predisposition. Signalment, presence of endocrine disease, clinicopathologic results, and outcome were compared between the BT, other breeds, and control BTs. RESULTS: The odds of identifying a GBM in a BT in this hospital population was 85 times that of all other breeds (95% confidence interval 56.9-126.8). BT had similar clinical signs and clinicopathologic changes to other breeds with GBM. There was no evidence that endocrinopathies were associated with GBM in BT. CLINICAL SIGNIFICANCE: A robust breed predisposition to GBM is established for the BT.

25-Hydroxy vitamin D3 serum concentration in dogs with acute polyradiculoneuritis compared to matched controls.

OBJECTIVES: To determine if dogs with acute polyradiculoneuritis have lower serum 25-hydroxy vitamin D3 concentration compared to a control group of dogs with idiopathic epilepsy. MATERIALS AND METHODS: Retrospective case-control study of 21 dogs with acute canine polyradiculoneuritis and 21 control dogs with idiopathic epilepsy matched for year and season of presentation from a referral hospital population in the UK. Serum concentration of 25-hydroxy vitamin D3 was compared between groups using Student's t-test. RESULTS: Dogs with acute canine polyradiculoneuritis had significantly lower (P=0·033) serum 25-hydroxy vitamin D3 concentration (87·1 nmol/L ±55·4 nmol/L) compared to a control group with idiopathic epilepsy (113 nmol/L ±66·3 nmol/L). CLINICAL SIGNIFICANCE: The cause and clinical significance of the altered vitamin D status in dogs with acute polyradiculoneuritis are not clear and require further investigation. Our findings pave the way for improved understanding of acute canine polyradiculoneuritis and, potentially, improved clinical management, if a causal role for 25-hydroxy vitamin D3 is defined.

Whole-Blood Taurine Concentrations in Cats With Intestinal Disease.

BACKGROUND: Increased delivery of taurine-conjugated bile acids to the distal bowel can lead to dysbiosis resulting in colitis in mouse models of inflammatory bowel disease. A similar situation also could occur in cats with intestinal disease and might therefore result in decreased whole-body taurine concentration. HYPOTHESIS/OBJECTIVES: To determine whether whole-blood taurine concentrations are decreased at the time of diagnosis in cats with intestinal disease and to correlate concentrations with clinical and laboratory variables. ANIMALS: Twenty-one cats with chronic inflammatory enteropathy and 7 cats with intestinal neoplasia from the University of Bristol. METHODS: Cats that had undergone a thorough investigation consisting of a CBC, serum biochemistry, serum cobalamin and folate concentrations, transabdominal ultrasound examination and histopathology of intestinal biopsy specimens, as well as additional testing if indicated, were included. Whole-blood from these cats collected at the time of histologic diagnosis and stored in ethylenediaminetetraacetic acid was retrospectively analyzed for taurine with an automated high-performance liquid chromatography amino acid analyzer. RESULTS: Although whole-blood taurine concentrations remained within the reference range, those cats with predominantly large intestinal clinical signs had significantly lower concentrations than did cats with small intestinal and mixed bowel clinical signs (P = 0.033) and this difference also was significant when assessed only in cats with chronic inflammatory enteropathy (P = 0.019). CONCLUSIONS AND CLINICAL IMPORTANCE: Additional studies are needed to determine whether large intestinal signs in cats with chronic inflammatory enteropathy are caused by alterations in the microbiota arising as a consequence of increased delivery of taurine-conjugated bile acids.

Lack of correlation between mucosal immunoglobulin A-positive plasma cell numbers and TLR5 genotypes in German shepherd dogs with idiopathic chronic enteropathy.

It has been suggested previously that a deficiency in mucosal immunoglobulin (Ig) A production could be involved in the pathogenesis of chronic enteropathy in German shepherd dogs (GSDs). Recent research has shown that single nucleotide polymorphisms in the gene encoding Toll-like receptor (TLR)-5 are associated with an increased risk of development of chronic idiopathic enteropathy in this breed. IgA is essential for mucosal immunity and studies in mice have linked the interaction of TLR5 with its ligand flagellin to class switching of B cells into IgA-producing plasma cells. We hypothesized that dogs carrying the risk-associated (RA) genotypes for G22A and C100T genes of TLR5 would have a different number of IgA plasma cells in the duodenal and colonic mucosa compared with dogs carrying the risk-protective (RP) genotypes. Thirty-one GSDs were diagnosed with idiopathic chronic enteropathy by clinical exclusion diagnosis and histopathological confirmation. Immunohistochemistry was performed using goat anti-dog IgA primary antibody. Two sections of duodenum, and colon if available, were examined from each animal. Twelve images were captured from each section and IgA-positive cells were counted and expressed per 10,000 µm(2). TLR5 genotypes for the G22A and C100T genes were determined by polymerase chain reaction on blood samples. Numbers of IgA-positive cells in the duodenum and colon were slightly higher than those published previously for GSDs with or without chronic enteropathy (mean in the crypt area of the duodenum 52.6 ± 16.2; mean in the tip of the duodenal villus 51.12 ± 3.83; mean in the base of the duodenal villus 55.02 ± 3.3; mean in the crypt area of the colon 67.4 ± 4.3). There was no correlation between numbers of IgA-positive cells in duodenum or colon between dogs carrying the RA versus the RP alleles of TLR genes. Further studies are needed to assess the production of secretory IgA and its relationship to TLR5 genotypes.

Association between nucleotide oligomerisation domain two (Nod2) gene polymorphisms and canine inflammatory bowel disease.

The most important genetic associations that have been implicated to play a role in the etiology of Crohn's disease (CD) in humans are single nucleotide polymorphisms (SNPs) in nucleotide oligomerisation domain 2 (NOD2). The aim of this study was to investigate whether SNPs in the canine NOD2 gene are associated with inflammatory bowel disease (IBD) in German shepherd dogs (GSDs) and other canine breeds. A mutational analysis of the NOD2 gene was carried out in 10 randomly selected GSDs with IBD. The mutational analysis identified five non-synonymous SNPS, of which four in exon 3 of the NOD2 gene were evaluated in a case-control study using sequence based typing. Sequencing information from 55 GSDs with IBD were compared to a control group consisting of 61 GSDs. In addition, 85 dogs of other breeds with IBD and a breed-matched control group consisting of 162 dogs were also genotyped. All four SNPs were in complete linkage and, in the GSD population, were found to be in Hardy-Weinberg equilibrium. When the GSD case population was compared to the GSD control group, the heterozygote genotype for all four SNPs was more frequently found in the IBD population (p=0.03, OR=2.30, CI=1.07-4.94). However, these results were not mirrored in other canine breeds. Our study suggests that the four SNPs in exon 3 of NOD2 are significantly associated with IBD in GSDs when analyzed in an over-dominant model. However, these results were not mirrored in other canine breeds with IBD. This suggests that the etiology of this disease is complex and may involve the interaction of SNPs present in several genes or pathways to bring about the inflammatory changes seen in the intestine.

Canine breeds at high risk of developing inflammatory bowel disease in the south-eastern UK.

Genetics are an important factor in the development of human inflammatory bowel disease (IBD); however, there is very little information available regarding the role of genetics in canine IBD. The purpose of this study was to gather information about which canine breeds in the south-eastern UK are at a high risk for developing IBD. Determination of such breeds may help further genetic research in this complex disease. The computer medical records at the Queen Mother Hospital for Animals, Royal Veterinary College dating from August 1, 2003 to December 31, 2009 were retrospectively searched for cases diagnosed with IBD. Five hundred and forty-six dogs with IBD were identified, representing 86 different breeds. The comparison group consisted of all dogs from these same 86 breeds without IBD admitted to the hospital during the same period that amounted to 27,463 dogs. The breeds at significantly higher risk of developing IBD compared with mixed-breed dogs consisted of weimaraner (odds ratio [OR]=3.6797, 95 per cent confidence interval [CI]=2.0167 to 6.7141, P<0.0001), rottweiler (OR=2.9697, 95 per cent CI=1.7569 to 5.0196, P<0.0001), German shepherd dog (GSD) (OR=2.4101, 95 per cent CI=1.5826 to 3.36705, P<0.0001), border collie (OR=1.9936, 95 per cent CI=1.1655 to 3.4101, P=0.0118) and boxer (OR=1.6961, 95 per cent CI=1.0441 to 2.755, P=0.0328). This study demonstrates for the first time canine breeds in the south-eastern UK that are highly susceptible to developing IBD. Identification of such breeds may allow for a more focused investigation of genetic mutations associated with canine IBD.

Breed-independent toll-like receptor 5 polymorphisms show association with canine inflammatory bowel disease.

Inflammatory bowel disease (IBD) is thought to be the most common cause of vomiting and diarrhoea in dogs. Although IBD can occur in any canine breed, certain breeds are more susceptible. We have previously shown that polymorphisms in the TLR4 and TLR5 (toll-like receptor) genes are significantly associated with IBD in German Shepherd dogs (GSDs). In order to allow for the development of novel diagnostics and therapeutics suitable for all dogs suffering from IBD, it would be useful to determine if the described polymorphisms are also significantly associated with IBD in other breeds. Therefore, the aim of this study was to investigate whether polymorphisms in the canine TLR4 and TLR5 genes are associated with IBD in other non-GSD canine breeds. The significance of the previously identified non-synonymous single nucleotide polymorphisms (SNPs) in the TLR4 (T23C, G1039A, A1571T and G1807A) and TLR5 genes (G22A, C100T and T1844C) were evaluated in a case-control study using a SNaPSHOT multiplex reaction. Sequencing information from 85 unrelated dogs with IBD consisting of 38 different breeds was compared with a breed-matched control group consisting of 162 unrelated dogs. Indeed, as in the GSD IBD population, the two TLR5 SNPs (C100T and T1844C) were found to be significantly protective for IBD in other breeds (P = 0.023 and P = 0.0195 respectively). Our study suggests that the two TLR5 SNPs, C100T and T1844C could play a role in canine IBD as these were found to be protective factors for this disease in 38 different canine breeds. Thus, targeting TLR5 in the canine system may represent a suitable way to develop new treatment for IBD in dogs.

CD11c+ cells are significantly decreased in the duodenum, ileum and colon of dogs with inflammatory bowel disease.

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<0.01, P<0.01 and P<0.05, respectively). There was a significant negative correlation between the number of CD11c(+) cells in the colon of dogs with IBD and the CCECAI (P=0.044, r(2)=-0.558). Chronic inflammation in canine IBD appears to involve an imbalance in the intestinal DC population. Future studies will determine whether reduced expression of CD11c could be a useful marker for the diagnosis and monitoring of canine IBD.

Elevated canine pancreatic lipase immunoreactivity concentration in dogs with inflammatory bowel disease is associated with a negative outcome.

OBJECTIVES: To investigate whether elevated canine pancreatic lipase immunoreactivity (CPLI) concentrations in dogs with inflammatory bowel disease (IBD) is associated with a worse clinical outcome. METHODS: Serum CPLI assays were performed on serum stored from cases diagnosed with IBD. Thirty-two dogs with CPLI results within the reference range were designated as the control group and 15 dogs had CPLI above the reference range. Clinical signs, age, serum lipase and amylase activities, serum albumin and cobalamin concentrations, abdominal ultrasound examination, histopathology on small intestinal biopsies, management of IBD and outcome were compared between the two groups. RESULTS: No significant differences were found in clinical activity score (P=0.54), number of antibiotic-responsive disease cases (P=0.480), number of steroid-responsive disease cases (P=0.491), serum amylase activity (P=0.058), serum cobalamin concentration (P=0.61), serum albumin concentration (P=0.052), abdominal ultrasound score (P=0.23) and histopathology scores for IBD (P=0.74) between the two groups. Dogs with increased CPLI concentration were significantly older and had a higher serum lipase activity than dogs with a CPLI concentration within the normal reference range (P=0.001, P=0.001, respectively). Moreover, dogs with increased CPLI concentration responded poorly to steroid treatment (P=0.01) and were significantly more likely to be euthanased at follow-up (P=0.02). CLINICAL SIGNIFICANCE: CPLI should be measured in cases of canine IBD as elevated CPLI was associated with a worse outcome.