Identification of Mucosa-Invading and Intravascular Bacteria in Feline Small Intestinal Lymphoma.

Persistent bacterial infections of the gastrointestinal mucosa are causally linked to gastric carcinoma and mucosa-associated lymphoid tissue (MALT) lymphoma in people and laboratory animals. We examined the relationship of mucosa-associated bacteria to alimentary lymphoma in cats. Intestinal biopsies from 50 cats with alimentary lymphoma (small cell, n = 33; large cell, n = 17) and 38 controls without lymphoma (normal to minimal change on histopathology, n = 18; lymphocytic-plasmacytic enteritis, n = 20) were evaluated. The number and spatial distribution of bacteria (ie, in luminal cellular debris, villus-associated mucus, adherent to epithelium, mucosal invasion, intravascular, or serosal) were determined by fluorescence in situ hybridization with the eubacterial probe EUB-338. Mucosa-invasive bacteria were more frequently observed in cats with large cell lymphoma (82%, P ≤ .001) than in cats with small cell lymphoma (18%), normal to minimal change on histopathology, and lymphocytic-plasmacytic enteritis (3%). Intravascular bacteria were observed solely in large cell lymphoma (29%), and serosal colonization was more common in cats with large cell lymphoma (57%) than with small cell lymphoma (11%, P ≤ .01), normal to minimal change (8%, P ≤ .01), and lymphocytic-plasmacytic enteritis (6%, P ≤ .001). The high frequency of invasive bacteria within blood vessels and serosa of cats with large cell lymphoma may account for the sepsis-related complications associated with large cell lymphoma and inform clinical management. Further studies are required to determine the role of intramucosal bacteria in the etiopathogenesis of feline alimentary lymphoma.

Equine stomachs harbor an abundant and diverse mucosal microbiota.

Little is known about the gastric mucosal microbiota in healthy horses, and its role in gastric disease has not been critically examined. The present study used a combination of 16S rRNA bacterial tag-encoded pyrosequencing (bTEFAP) and fluorescence in situ hybridization (FISH) to characterize the composition and spatial distribution of selected gastric mucosal microbiota of healthy horses. Biopsy specimens of the squamous, glandular, antral, and any ulcerated mucosa were obtained from 6 healthy horses by gastroscopy and from 3 horses immediately postmortem. Pyrosequencing was performed on biopsy specimens from 6 of the horses and yielded 53,920 reads in total, with 631 to 4,345 reads in each region per horse. The microbiome segregated into two distinct clusters comprised of horses that were stabled, fed hay, and sampled at postmortem (cluster 1) and horses that were pastured on grass, fed hay, and biopsied gastroscopically after a 12-h fast (cluster 2). The types of bacteria obtained from different anatomic regions clustered by horse rather than region. The dominant bacteria in cluster 1 were Firmicutes (>83% reads/sample), mainly Streptococcus spp., Lactobacillus spp. and, Sarcina spp. Cluster 2 was more diverse, with predominantly Proteobacteria, Bacteroidetes, and Firmicutes, consisting of Actinobacillus spp. Moraxella spp., Prevotella spp., and Porphyromonas spp. Helicobacter sp. sequences were not identified in any of 53,920 reads. FISH (n = 9) revealed bacteria throughout the stomach in close apposition to the mucosa, with significantly more Streptococcus spp. present in the glandular region of the stomach. The equine stomach harbors an abundant and diverse mucosal microbiota that varies by individual.

Clinical characteristics and long-term outcome of E. coli-associated granulomatous ileocolitis in dogs: five cases (2010-2014).

OBJECTIVES: To describe the clinical characteristics and long-term outcome of Escherichia coli-associated granulomatous ileocolitis in dogs. METHODS: Retrospective review of medical records from dogs with periodic acid-Schiff positive (PAS+) granulomatous ileocolitis and mucosally invasive E. coli in the ileum and colon. Initial bacterial colonisation was evaluated using fluorescence in situ hybridization (FISH) in all dogs and corroborated with colonic and/or ileal culture, when performed. RESULTS: Four boxer dogs and 1 French Bulldog with PAS+ granulomatous ileocolitis (GIC) were evaluated. All dogs had chronic diarrhoea refractory to empirical therapy. Ileocolonoscopy revealed mucosal haemorrhage and ulceration in the ileum (3/4) and colon (5/5). E. coli were visualised as clusters within the ileal and colonic mucosa. Complete (CR, 4/5) or partial (PR, 1/5) clinical response to fluoroquinolones was noted in all dogs within 30 days. CR was sustained in three of four dogs (median disease-free interval 40 months, range 16 to 60). Two dogs relapsed while receiving fluoroquinolones. Repeat biopsy isolated multidrug-resistant, mucosally invasive E. coli in the ileum (1/2) and colon (2/2). Targeted antimicrobial therapy was associated with long-term PR (78 months) in both dogs. CLINICAL SIGNIFICANCE: Concurrent E. coli-associated granulomatous inflammation in the ileum and colon did not impart a poor clinical outcome or lack of response to the conventional standard of care for granulomatous colitis in dogs that were aggressively diagnosed and treated. Clinical outcome was influenced by antimicrobial resistance, with response dependent upon antimicrobial therapy informed by susceptibility testing.

Comparison of 4 Giardia diagnostic tests in diagnosis of naturally acquired canine chronic subclinical giardiasis.

BACKGROUND: The performance of Giardia diagnostic tests that detect either cysts or fecal antigens has not been thoroughly examined. HYPOTHESIS/OBJECTIVES: We examined the concordance and agreement among 4 Giardia diagnostic tests (2 cyst and 2 coproantigen detection methods) in a colony of dogs chronically and subclinically infected with Giardia. ANIMALS: Twenty dogs with chronic, subclinical Giardia infection. METHODS: Giardia diagnostic tests were performed repeatedly on each dog over 120 days. Fecal cyst detection methods (ZnSO4 flotation and fluorescent antibody [FAB] coproscopy) were performed 3 times per week. Coproantigen methods (Giardia SNAP test and quantitative ELISA) were performed weekly. Results were analyzed and compared among methods. RESULTS: When compared with FAB coproscopy, all of the in-house diagnostic tests had excellent positive predictive values (PPVs, 95-99%) at the study prevalence (89%). At lower prevalence rates, ZnSO4, SNAP, and ELISA tests all had good negative predictive values (NPVs), but poor PPVs. There was poor to good agreement among tests by kappa analysis. CONCLUSION AND CLINICAL RELEVANCE: Our findings show that most commonly used in-house Giardia diagnostic tests have poor agreement with the gold standard method (FAB coproscopy). The in-house tests have good NPVs, but poor PPVs, at prevalence rates common in most clinical settings.

Antimicrobial resistance impacts clinical outcome of granulomatous colitis in boxer dogs.

BACKGROUND: Escherichia coli have recently been identified within the colonic mucosa of Boxer dogs with granulomatous colitis (GC). Eradication of invasive E. coli is associated with clinical and histological remission. OBJECTIVES: To determine antimicrobial susceptibility profiles of E. coli strains from GC and healthy dogs, and the association of antimicrobial resistance with clinical outcome. ANIMALS: Fourteen Boxer dogs with GC and 17 healthy pet dogs. METHODS: Prospective study: E. coli was cultured from GC biopsies and rectal mucosal swabs of healthy dogs. Individual strains were selected by phylogroup and overall genotype, determined by triplex- and random amplified polymorphic DNA-polymerase chain reaction respectively. Antimicrobial susceptibility was determined by broth microdilution minimal inhibitory concentration. RESULTS: Culture yielded 23 E. coli strains from GC (1-3/dog, median 2) and 34 strains from healthy (1-3/dog, median 2). E. coli phylogroups were similar (P=.18) in GC (5A, 7B1, 5B2, 6D) and healthy (2A, 10B1, 15B2, 7D). Resistance to ampicillin, amoxicillin-clavulanate, cefoxitin, tetracycline, trimethoprim-sulfa (TMS), ciprofloxacin, and chloramphenicol was greater (P<.05) in GC (21-64%) than healthy (0-24%). Enrofloxacin resistant E. coli were isolated from 6/14 GC versus 0/17 healthy (P=.004). Of the enrofloxacin resistant cases, 4/6 were also resistant to macrophage-penetrating antimicrobials such as chloramphenicol, rifampicin, and TMS. Enrofloxacin treatment before definitive diagnosis was associated with antimicrobial resistance (P<.01) and poor clinical outcome (P<.01). CONCLUSIONS AND CLINICAL IMPORTANCE: Antimicrobial resistance is common among GC-associated E. coli and impacts clinical response. Antimicrobial therapy should be guided by mucosal culture and antimicrobial susceptibility testing rather than empirical wisdom.

Influence of Enterococcus faecium SF68 probiotic on giardiasis in dogs.

BACKGROUND: Giardiasis is a common, potentially zoonotic disease, and dogs often harbor and shed cysts without showing clinical signs. Treatment with the probiotic Enterococcus faecium SF68 has been shown to stimulate mucosal and systemic immunity in a variety of animal models and in young dogs, and to reduce giardial cyst and antigen shedding in rodents. HYPOTHESIS: Adult dogs with chronic naturally acquired giardiasis will have decreased giardial fecal cyst and antigen shedding and increased innate and adaptive immunity after 6 weeks probiotic treatment with E. faecium SF68. ANIMALS: Twenty adult dogs. METHODS: After a 6-week dietary equilibration period, dogs were randomized to receive E. faecium SF68 or placebo for 6 weeks, and then crossed over to the alternate treatment. We measured cyst shedding, fecal giardial antigen, fecal immunoglobulin A (IgA) concentration, and circulating leukocyte phagocytic activity at multiple timepoints to determine the effect of E. faecium SF68 on giardiasis and immune responses in these dogs. RESULTS: No differences were observed between placebo or E. faecium SF68 treatment for giardial cyst shedding, fecal antigen shedding, fecal IgA concentration, or leukocyte phagocytic activity. CONCLUSIONS: Short-term treatment with E. faecium SF68 of dogs with chronic naturally acquired subclinical giardiasis fails to affect giardial cyst shedding or antigen content and does not alter innate or adaptive immune responses.

Fluorescence in situ hybridization confirms clearance of visible Helicobacter spp. associated with gastritis in dogs and cats.

BACKGROUND: The results of studies examining the role of Helicobacter spp. in the pathogenesis of canine and feline gastritis are inconclusive. Furthermore, data evaluating the effectiveness of medical therapy for eradication of Helicobacter infection are limited. AIM: To detect Helicobacter spp. in mucosal biopsies of dogs and cats diagnosed with gastritis, with fluorescence in situ hybridization (FISH). ANIMALS: Three dogs and 2 cats with signs of chronic gastrointestinal disease. METHODS: Dogs and cats infected with Helicobacter spp. were treated with triple antimicrobial therapy and fed an elimination diet for 21 days. Helicobacter spp. status in endoscopic (3 dogs, 1 cat) or surgical biopsies (1 cat) of gastric mucosa was compared pre- and posttreatment in each animal by histology, FISH analysis, and polymerase chain reaction (PCR). RESULTS: Gastritis of varying severity with intraglandular spiral bacteria was observed in all animals. Pretreatment diagnostic tests confirmed the presence of mucosal Helicobacter spp. in all animals by FISH and histopathology and in 4/5 animals by PCR. Rapid resolution of vomiting episodes was observed in all animals. Gastric biopsies performed after triple therapy revealed clearance of visible Helicobacter spp. by histopathology and negative FISH analysis, as well as PCR in all animals. CONCLUSIONS AND CLINICAL IMPORTANCE: Application of FISH to routine biopsy specimens enabled rapid and specific identification of Helicobacter spp. within the gastric mucosa of dogs and cats. Although medical therapy was useful in resolution of clinical signs and clearance of visible Helicobacter spp. in gastric biopsies, gastric inflammation persisted.

Remission of histiocytic ulcerative colitis in Boxer dogs correlates with eradication of invasive intramucosal Escherichia coli.

BACKGROUND: Historically, histiocytic ulcerative (HUC) (or granulomatous) colitis of Boxer dogs was considered an idiopathic immune-mediated disease with a poor prognosis. Recent reports of dramatic responses to enrofloxacin and the discovery of invasive Escherichia coli within the colonic mucosa of affected Boxer dogs support an infectious etiology. HYPOTHESIS: Invasive E. coli is associated with colonic inflammation in Boxer dogs with HUC, and eradication of intramucosal E. coli correlates with clinical and histologic remission. ANIMALS: Seven Boxer dogs with HUC. METHODS: Prospective case series. Colonic biopsies were obtained at initial evaluation in 7 dogs, and in 5 dogs after treatment with enrofloxacin. Biopsies were evaluated by standardized histopathology, and fluorescence in situ hybridization (FISH) with probes to eubacteria and E. coli. RESULTS: Intramucosal E. coli was present in colonic biopsies of 7/7 Boxers with HUC. Clinical response was noted in all dogs within 2 weeks of enrofloxacin (7 + or - 3.06 mg/kg q24 h, for 9.5 + or - 3.98 weeks) and was sustained in 6 dogs (median disease-free interval to date of 47 months, range 17-62). FISH was negative for E. coli in 4/5 dogs after enrofloxacin. E. coli resistant to enrofloxacin were present in the FISH-positive dog that relapsed. CONCLUSIONS AND CLINICAL RELEVANCE: The correlation between clinical remission and the eradication of mucosally invasive E. coli during treatment with enrofloxacin supports the causal involvement of E. coli in the development of HUC in susceptible Boxer dogs. A poor response to enrofloxacin treatment might be due to colonization with enrofloxacin-resistant E. coli.

Eubacterial fluorescence in situ hybridisation and histologic features in 25 dogs with gallbladder mucocele.

OBJECTIVES: To detect and localise bacteria in gallbladder mucoceles using fluorescence in situ hybridisation (FISH). To report clinical signs, clinicopathologic abnormalities, sonographic findings and histopathological findings in FISH+ and FISH- dogs with gallbladder mucoceles. MATERIALS AND METHODS: Retrospective review of signalment, clinical signs, clinicopathologic and sonographic findings of 25 cases of histopathologically confirmed gallbladder mucocele. Histopathological sections of gallbladder mucocele were evaluated for cystic mucinous hyperplasia, cystic mucinous hyperplasia with cholecystitis and rupture. The number and spatial distribution of bacteria was determined by eubacterial FISH. Gallbladder contents were cultured in 21 dogs. RESULTS: Bacteria were detected within or adherent to the gallbladder wall in eight of 25 (32%) cases. Bacterial culture was positive in one dog. Cystic mucinous hyperplasia with concurrent cholecystitis was found in 17 of 25 (68%) of dogs with gallbladder mucocele. CLINICAL SIGNIFICANCE: FISH was more sensitive for detection of bacteria in gallbladder mucoceles when compared to bacterial culture of bile. Cholecystitis was common in dogs with gallbladder mucocele. Further study is required to elucidate the relationship of cystic mucinous hyperplasia, bacteria and cholecystitis in the aetiopathogenesis and progression of gallbladder mucocele.

The relationship of mucosal bacteria to duodenal histopathology, cytokine mRNA, and clinical disease activity in cats with inflammatory bowel disease.

Feline inflammatory bowel disease (IBD) is the term applied to a group of poorly understood enteropathies that are considered a consequence of uncontrolled intestinal inflammation in response to a combination of elusive environmental, enteric microbial, and immunoregulatory factors in genetically susceptible cats. The present study sought to examine the relationship of mucosal bacteria to intestinal inflammation and clinical disease activity in cats with inflammatory bowel disease. Duodenal biopsies were collected from 27 cats: 17 undergoing diagnostic investigation of signs of gastrointestinal disease, and 10 healthy controls. Subjective duodenal histopathology ranged from normal (10), through mild (6), moderate (8), and severe (3) IBD. The number and spatial distribution of mucosal bacteria was determined by fluorescence in situ hybridization with probes to 16S rDNA. Mucosal inflammation was evaluated by objective histopathology and cytokine profiles of duodenal biopsies. The number of mucosa-associated Enterobacteriaceae was higher in cats with signs of gastrointestinal disease than healthy cats (P<0.001). Total numbers of mucosal bacteria were strongly associated with changes in mucosal architecture (P<0.001) and the density of cellular infiltrates, particularly macrophages (P<0.002) and CD3(+)lymphocytes (P<0.05). The number of Enterobacteriaceae, E. coli, and Clostridium spp. correlated with abnormalities in mucosal architecture (principally atrophy and fusion), upregulation of cytokine mRNA (particularly IL-1, -8 and -12), and the number of clinical signs exhibited by the affected cats. These data establish that the density and composition of the mucosal flora is related to the presence and severity of intestinal inflammation in cats and suggest that mucosal bacteria are involved in the etiopathogenesis of feline IBD.

Search for Borrelia burgdorferi in kidneys of dogs with suspected "Lyme nephritis".

BACKGROUND: "Lyme nephritis" is a poorly characterized condition associated with proteinuria and often fatal renal failure in dogs with serological evidence of infection with Borrelia burgdorferi. OBJECTIVE: The aim of this study was to determine if intact B. burgdorferi organisms were present in the kidneys of serologically Lyme-positive dogs with histopathologic features of Lyme nephritis. ANIMALS: Twenty-six affected and 10 control dogs were identified over an 8-year period (1996-2004) in databases at Cornell University's College of Veterinary Medicine. Case inclusion required serologic evidence of natural exposure to B. burgdorferi and availability of renal tissue (frozen or paraffin embedded) exhibiting pathology consistent with Lyme nephritis. METHODS: Renal tissue samples were assessed using modified Steiner (silver) (MS) staining, immunohistochemistry (IHC), polymerase chain reaction (PCR) using 4 primer sets (eubacterial, B. burgdorferi, Bartonella, and canine genomic DNA), and fluorescence in situ hybridization (FISH) using a 5'-cy3-eubacterial probe for 16S rRNA. RESULTS: MS stain was positive in 1 case; IHC was negative in all cases. None of the B. burgdorferi or Bartonella PCR reactions was positive. Two of the B. burgdorferi FISH analyses were positive. CONCLUSIONS AND CLINICAL IMPORTANCE: Minimal evidence of the presence of intact B. burgdorferi or any other bacterial organism was found in the renal tissue of dogs with suspected Lyme nephritis. Direct renal invasion by B. burgdorferi organisms does not appear to be responsible for this syndrome.

Cardiac troponin I is elevated in dogs and cats with azotaemia renal failure and in dogs with non-cardiac systemic disease.

OBJECTIVE: To determine if dogs and cats with renal failure, or other severe non-cardiac disease, and no antemortem evidence of cardiac disease on basic clinical evaluation, have elevated levels of cardiac troponin I (cTnI). DESIGN: Cross-sectional study using 56 dogs and 14 cats with primary non-cardiac disease (39 dogs with azotaemic renal failure, 14 cats with azotaemic renal failure, 17 dogs with non-cardiac systemic disease); 7/25 dogs and 6/14 cats had murmurs detected on physical examination. Serum or heparinised plasma was collected and analysed for cTnI. RESULTS: Cardiac troponin I concentrations were elevated above reference intervals in 70% of dogs and 70% of cats with azotaemic renal failure and in 70% of dogs with a variety of systemic non-cardiac diseases. Cardiac troponin I concentrations did not correlate with the degree of azotaemia, presence of murmurs, hypertension or type of non-cardiac illness. CONCLUSIONS: Cardiac troponin I concentration is often elevated in dogs and cats with azotaemic renal failure and in dogs with other systemic non-cardiac illness, suggesting that these conditions often result in clinically inapparent myocardial injury or possibly altered elimination of cTnI.

Association of Helicobacter with cholangiohepatitis in cats.

Infection with Helicobacter spp. is increasingly linked with hepatobiliary inflammation and neoplasia in people and in a variety of animals. We sought to determine if Helicobacter species infection is associated with cholangiohepatitis in cats. Deoxyribonucleic acid was extracted from tissue blocks from cats with cholangiohepatitis (32), noninflammatory liver disease (13), and cats with normal liver histology (4). Deoxyribonucleic acid was polymerase chain reaction-amplified with 2 sets of Helicobacter genus-specific primers, gel purified, and sequenced. Polymerase chain reaction-positive hepatic tissue was further examined with Steiner's stain, immunocytochemistry for Helicobacter species, and eubacterial fluorescent in situ hybridization. Gastric tissues of cats with known Helicobacter infection status served as controls for deoxyribonucleic acid extraction and sequence comparison. Helicobacter species were detected in 2/32 cats with cholangiohepatitis, and 1/17 controls. Sequences had 100% identity with Helicobacter species liver, Helicobacter pylori, and Helicobacter fenelliae/cinaedii in a cat with suppurative cholangitis, Helicobacter species liver, Helicobacter pylori, and Helicobacter nemistrineae in a cat with mild lymphocytic portal hepatitis, and Helicobacter bilis in a cat with portosystemic vascular anomaly. In contrast, sequences from gastric biopsies showed highest homology (99-100%) to "Helicobacter heilmannii," Helicobacter bizzozeronii, Helicobacter felis, and Helicobacter salomonis. Fluorescent in situ hybridization revealed a semicurved bacterium, with Helicobacter-like morphology, in an intrahepatic bile duct of the cat with suppurative cholangitis. This study has identified Helicobacter deoxyribonucleic acid in 2/32 cats with cholangiohepatitis and 1/13 cats with noninflammatory liver disease. Deoxyribonucleic acid sequences of hepatic Helicobacter species were distinct from those found in the stomach and are broadly consistent with those identified in cat intestine and bile, and hepatobiliary disease in people and rodents.

Evaluation of capsule endoscopy to detect mucosal lesions associated with gastrointestinal bleeding in dogs.

OBJECTIVE: The objective of this study was to examine the utility of capsule endoscopy to detect mucosal abnormalities in dogs with gastrointestinal haemorrhage. METHODS: Capsules were administered to 2 healthy controls and 8 patients with gastrointestinal haemorrhage. Images were evaluated for quality, gastric emptying time, small intestinal transit time and presence of lesions. RESULTS: There were no adverse effects of capsule endoscopy in dogs weighing from 7·7 to 58 kg. The capsule traversed the entire gastrointestinal tract in 5 of 8 patients, with high quality images obtained in the stomach and small intestine. Gastric emptying time and small intestinal transit time ranged from 1 to 270 and 15 to 180 minutes, respectively. In 3 of 8 patients, the capsule remained in the stomach despite pro-kinetics. Gastric lesions included mild haemorrhage and pinpoint erosion (4 of 8), a mass (1) and thickened bleeding pyloric mucosa (2). Two of 3 dogs with capsule retention had gastric lesions. Intestinal lesions included a healing duodenal ulcer, abnormal villi, ileal ulceration and colonic bleeding. Lesions identified by capsule endoscopy were considered a significant source of haemorrhage in 4 of 7 dogs with active bleeding. The relevance of pinpoint gastric mucosal erosions to blood loss is unclear. CLINICAL SIGNIFICANCE: Capsule endoscopy can enable the non-invasive detection of gastric and small intestinal mucosal lesions in dogs presenting for evaluation of gastrointestinal bleeding.

Focal intestinal lipogranulomatous lymphangitis in 10 dogs.

OBJECTIVES: To describe the clinical and pathological features of canine focal lipogranulomatous lymphangitis, to evaluate its underlying infectious cause and to compare it with human Crohn's disease. METHODS: Retrospective review of case records with a histopathological diagnosis of focal lipogranulomatous lymphangitis. Bacterial and fungal colonisation was evaluated using fluorescence in situ hybridisation and histochemical staining, respectively. A comparison with Crohn's disease was performed by a human pathologist. RESULTS: Ten dogs were evaluated. The historical complaints were predominantly chronic diarrhoea (10/10) and vomiting (5/10). The biochemical abnormalities included hypoalbuminaemia (6/10) and hypocobalaminaemia (4/6). Abdominal sonography revealed a thickened distal ileum±ileocolic junction. Colonoscopy showed a swollen caecal ostium and oedematous caecum in 7/10 dogs. A stenotic ileo-colic opening prevented endoscopic intubation in all dogs. Histology from the resected lesions revealed granulomatous inflammation involving the muscularis and serosa. Fluorescence in situ hybridization demonstrated invasive bacteria in 2/10 dogs. Post-resection, all dogs received metronidazole and tapering immunosuppressive doses of prednisolone. Remission (median 17 months) was achieved in 8/10 dogs. CLINICAL SIGNIFICANCE: Focal lipogranulomatous lymphangitis is a rare and severe form of canine inflammatory bowel disease with preferential localisation to the ileum and the ileocolic junction. An underlying infectious aetiology was not identified.

Pancreatitis and triaditis in cats: causes and treatment.

Pancreatitis in cats is frequently accompanied by concurrent disease in other organ systems. Co-morbidities include hepatic lipidosis, inflammatory liver disease, bile duct obstruction, diabetes mellitus, inflammatory bowel disease, vitamin deficiency (B12/cobalamin, folate or K), intestinal lymphoma, nephritis, pulmonary thromboembolism and pleural and peritoneal effusions. "Triaditis" is the term used to describe concurrent inflammation of the pancreas, liver and small intestines. Triaditis has been reported in 50 to 56% of cats diagnosed with pancreatitis and 32 to 50% of those with cholangitis/inflammatory liver disease. A definitive diagnosis of triaditis is based on the histopathological evaluation of each organ. However, the specific conditions of each organ that constitute a diagnosis of triaditis remains to be defined. While the aetiopathogenesis of pancreatitis and its relationship to inflammation in other organ systems is unclear, preliminary studies point to a heterogeneous group of conditions with differential involvement of host inflammatory and immune responses and enteric bacteria. Comprehensive, prospective studies that simultaneously evaluate the presence of predefined clinical, clinicopathological and histopathological abnormalities, coupled with high-resolution evaluation of pancreaticobiliary morphology, immunological profiling and screening for bacterial colonisation are required to advance diagnosis and therapy.

Helicobacter spp. infection in cats: evaluation of the humoral immune response and prevalence of gastric Helicobacter spp.

The principal aims of this study were to evaluate the humoral immune response (IgG) of cats with gastric Helicobacter spp. infection, and to determine the prevalence of different types of Helicobacter spp. in the stomachs of cats. The Helicobacter infection status of 45 cats (12 healthy spay/neuter cats, 9 sick cats, 24 colony cats) was determined by evaluating endoscopic gastric biopsies for urease activity, presence of Helicobacter-like organisms (HLO) on histopathology, and genus and species-specific PCR. Serum samples were evaluated with a kinetic enzyme linked immunosorbent assay (ELISA) utilizing the high molecular cell-associated protein (HM-CAP) fraction of H. felis ATCC 49179.Seventeen of 45 cats were infected with Helicobacter spp.: "H. heilmannii" 9/17, H. felis 4/17, mixed "H. heilmannii" and H. felis 3/17, unclassified-Helicobacter spp. 7/17. H. pylori was not detected in any cat. Kinetic ELISA results were significantly higher for infected cats, than for uninfected cats. Cats infected with different Helicobacter spp. showed similar distribution of OD/min values. There were no effects of age or clinical signs on the results of kinetic ELISA. No correlation between colonization density and seroconversion was observed. There were statistically significant, but weak correlations between the degree of seroconversion and the degree of inflammation, and the number of lymphoid follicles. Infected cats had more severe inflammation in the pylorus and fundus than uninfected cats. Infected sick cats had a higher degree of pyloric, but not fundic inflammation, than healthy infected cats and uninfected sick cats. The results indicate that naturally acquired infection with gastric Helicobacter spp. is associated with seroconversion (IgG) in cats. The similar ELISA values in cats infected with a variety of Helicobacter spp. suggests substantial antigenic homology between different Helicobacter spp. The higher degree of inflammation in infected than uninfected cats, supports a role for Helicobacter as a cause of gastritis in cats.

Characterization of microvillar membrane proteins of dog small intestine by two-dimensional electrophoresis.

A method for analysing microgram amounts of microvillar membranes by two-dimensional electrophoresis (protein mapping) is described, and has been used to characterize the microvillar proteins of the small intestine of German shepherd, corgi, and beagle dogs. Detergent-solubilized microvillar membranes were radiolabelled with 14C and separated by isoelectric focussing followed by SDS-PAGE. Proteins were detected fluorographically and glycoproteins by lectin-affinity staining. The microvillar hydrolases alkaline phosphatase and dipeptidyl aminopeptidase IV were identified by active-site labelling and aminopeptidase N by immunoprecipitation. Changes following pancreatic duct diversion were consistent with accumulation of pro-sucrase-isomaltase and diminished expression of the sucrase and isomaltase subunits. Cytoskeletal proteins were concentrated in the core fraction remaining after extraction of microvillar membranes with Triton X-100. There were no consistent differences between dogs of different breed, and the canine protein maps were similar to the human.

Histological and immunohistochemical detection of different Helicobacter species in the gastric mucosa of cats.

Detailed histopathological evaluation of the gastric mucosa of Helicobacter-infected cats is complicated by the difficulty of recognizing Helicobacter organisms on hematoxylin and eosin (HE)-stained sections and the ability of multiple Helicobacter species to infect cats. In this study, the presence and localization of different species of Helicobacter in the stomachs of cats was investigated using silver staining and immunohistochemistry. Five groups containing 5 cats each were established (group 1: urease negative and Helicobacter free; groups 2, 3, 4, and 5: urease positive and infected with Helicobacter heilmannii, unclassified Helicobacter spp., Helicobacter felis, and Helicobacter pylori, respectively). Gastric samples were evaluated by HE and silver staining and by immunohistochemistry with 3 different anti-Helicobacter primary antibodies. Helicobacter were detected by Steiner stain in all infected cats at the mucosal surface, in the lumen of gastric glands, and in the cytoplasm of parietal cells. In silver-stained sections, H. pylori was easily differentiated from H. felis, H. heilmannii, and unclassified Helicobacter spp., which were larger and more tightly coiled. No organisms were seen in uninfected cats. Helicobacter antigen paralleled the distribution of organisms observed in Steiner-stained sections for 2 of the 3 primary antibodies tested. The antisera were not able to discriminate between the different Helicobacter species examined. A small amount of Helicobacter antigen was present in the lamina propria of 3 H. pylori-, 3 H. felis-, and 1 H. heilmannii-infected cat. Minimal mononuclear inflammation was present in uninfected cats and in those infected with unclassified Helicobacter spp. and H. heilmannii cats. In H. felis-infected cats, lymphoid follicular hyperplasia with mild pangastric mononuclear inflammation and eosinophilic infiltrates were present. The H. pylori-infected cats had severe lymphoid follicular hyperplasia and mild to moderate mononuclear inflammation accompanied by the presence of neutrophils and eosinophils. These findings indicate that Steiner staining and immunohistochemistry are useful for detecting Helicobacter infections, particularly when different Helicobacter species can be present. Monoclonal antibodies specific for the different Helicobacter species could be important diagnostic aids. There appear to be differences in the severity of gastritis in cats infected with different Helicobacter species.

Hypercholesterolaemia in briards in the United Kingdom.

Elevated fasting plasma cholesterol concentrations were identified in clinically healthy briards. Biochemical investigations revealed no other major abnormalities. Plasma lipoprotein electrophoresis demonstrated a marked increase in the intensity of the alpha 2 band (compared with control dogs) which was reduced by dextran sulphate-magnesium chloride or sodium phosphotungstate-magnesium chloride precipitation of apo B and apo E containing lipoproteins in the plasma. The study has identified a hyperlipidaemia in briards characterised by increased cholesterol but normal triglyceride concentrations. The absence of obvious metabolic changes associated with secondary hypercholesterolaemia, suggests the breed may have a primary abnormality in cholesterol metabolism. The increased density of the precipitable lipoprotein which migrates to the alpha 2 band suggests that the hypercholesterolaemia may be due to an abnormal accumulation of high density lipoprotein (HDL) possibly HDLc. The possibility that abnormality in lipid metabolism might play a role in the development of retinal pigment epithelial dystrophy in briards is currently being investigated.