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Chronic hepatic disease can present a diagnostic challenge with different etiologies being associated with similar clinical and laboratory findings. The histopathological assessment of a liver biopsy specimen is usually required in order to make a definitive diagnosis and the availability of non-invasive prognostic biomarkers is limited. The emerging science of metabolomics is used to detect changes in endogenous low molecular weight metabolites in biological samples and offers the possibility of identifying noninvasive markers of disease. The objective of this study was to investigate differences in the urine metabolome between healthy dogs, dogs with chronic hepatitis, dogs with hepatocellular carcinoma, and dogs with a congenital portosystemic shunt. Stored urine samples from 10 healthy dogs, 10 dogs with chronic hepatitis, 6 dogs with hepatocellular carcinoma, and 5 dogs with a congenital portosystemic shunt were analyzed. The urine metabolome was analyzed by gas chromatography–quadrupole time of flight mass spectrometry and 220 known metabolites were identified. Principal component analysis and heat dendrogram plots of the metabolomics data showed clustering between groups. Random forest analysis showed differences in the abundance of various metabolites including putrescine, gluconic acid, sorbitol, and valine. Based on univariate statistics, 37 metabolites were significantly different between groups. In, conclusion, the urine metabolome varies between healthy dogs, dogs with chronic hepatitis, dogs with hepatocellular carcinoma, and dogs with a congenital portosystemic shunt. Further targeted assessment of these metabolites is needed to assess their diagnostic utility. [ABSTRACT FROM AUTHOR]

OBJECTIVE To develop and analytically validate a liquid chromatography-tandem mass spectrometry method for measurement of endogenous trans-4-hydroxy-L-proline concentrations in canine serum and to assess serum trans-4-hydroxy-L-proline concentrations in dogs with chronic hepatitis. SAMPLE Serum samples obtained from 20 dogs with histopathologically confirmed chronic hepatitis and 20 healthy control dogs. PROCEDURES A liquid chromatography-tandem mass spectrometry method for quantification of trans-4-hydroxy-L-proline concentration was developed and assessed for analytic sensitivity, linearity, accuracy, precision, and reproducibility. Serum concentration of trans-4-hydroxy-L-proline in dogs with chronic hepatitis and healthy control dogs was measured. RESULTS Observed-to-expected ratios for dilutional parallelism ranged from 72.7% to 111.5% (mean ± SD, 91.3 ± 19.6%). Intra-assay and interassay coefficients of variation ranged from 2.1% to 3.0% and 3.2% to 5.3%, respectively. Relative erro r ranged from -2.3% to 7.8%. Trans-4-hydroxy-L-proline concentrations were significantly lower in serum obtained from dogs with chronic hepatitis (median, 0.24 ng/mL; range, 0.06 to 1.84 ng/mL) than in serum obtained from healthy control dogs (median, 0.78 ng/mL; range, 0.14 to 4.90 ng/mL). CONCLUSIONS AND CLINICAL RELEVANCE The method described here for the quantification of trans-4-hydroxy-L-proline concentration in canine serum was found to be sensitive, specific, precise, accurate, and reproducible. Dogs with chronic hepatitis had significantly lower serum trans-4-hydroxy-L-proline concentrations than did healthy control dogs, possibly as a result of altered hepatic metabolism of amino acids. [ABSTRACT FROM AUTHOR]

Background: Chronic hepatopathies present a diagnostic challenge, with different diseases being associated with similar clinical and laboratory findings. Characterization of dogs with chronic hepatopathies can be difficult and require costly diagnostic procedures such as acquisition of a liver biopsy specimen. Noninvasive and inexpensive biomarkers that reliably characterize chronic hepatopathies such as chronic hepatitis or a congenital portosystemic vascular anomaly may decrease the need for costly or invasive diagnostic testing and guide novel therapeutic interventions. Objective: To investigate differences in the serum metabolome among healthy dogs, dogs with congenital portosystemic shunts, and dogs with chronic hepatitis. Animals: Stored serum samples from 12 healthy dogs, 10 dogs with congenital portosystemic shunts, and 6 dogs with chronic hepatitis were analyzed. Methods: The serum metabolome was analyzed with an untargeted metabolomics approach using gas chromatography–quadrupole time of flight mass spectrometry. Results: Principal component analysis and heat dendrogram plots of the metabolomics data showed clustering among individuals in each group. Random forest analysis showed differences in the abundance of various metabolites including increased aromatic amino acids and xylitol in dogs with congenital portosystemic shunts. Based on the univariate statistics, 50 metabolites were significantly different among groups. Conclusions and Clinical Importance: The serum metabolome varies among healthy dogs, dogs with congenital portosystemic shunts, and dogs with chronic hepatitis. Statistical analysis identified several metabolites that differentiated healthy dogs from dogs with vascular or parenchymal liver disease. Further targeted assessment of these metabolites is needed to confirm their diagnostic reliability. [ABSTRACT FROM AUTHOR]

Chronic hepatitis is the most common hepatic disease in dogs. Copper accumulation is an important cause of chronic hepatitis in dogs; however, the etiology in most dogs cannot be determined. Clinical signs of chronic hepatitis are often non-specific; therefore, this disease is frequently diagnosed in an advanced stage that makes successful intervention less likely. Early diagnosis of chronic hepatitis in dogs would thus be beneficial. The identification of proteins that are differentially expressed in dogs with chronic hepatitis could contribute to the development of novel diagnostic markers for this disease and provide insight into its pathogenesis. The objective of this study was to identify novel proteins that are differentially expressed in the liver of dogs with chronic hepatitis. Hepatic tissue was collected from 8 healthy dogs during ovariohysterectomy and from 8 dogs with histologically confirmed chronic hepatitis. The proteome of the liver samples was extracted by mechanical disruption and detergent-based cell lysis and differentially labeled prior to analysis by 2-dimensional fluorescence difference gel electrophoresis. Spots with an absolute fold change value > 2.0 were selected for further analysis. Protein identification was achieved by nanoflow liquid chromatography tandem mass spectrometry. Differential expression of select proteins was validated by Western blot. Five protein spots were differentially expressed between patients with chronic hepatitis and healthy control dogs. From these 5 protein spots 11 proteins were identified. Differential expression of cytokeratin 18 and annexin 5 were confirmed by Western blot analysis. Differential protein expression was shown between dogs with chronic hepatitis and healthy control dogs. Upregulation of cytokeratin 18 in chronic hepatitis may suggest increased hepatocellular apoptosis and necrosis, whereas upregulation of annexin 5A suggests increased hepatocellular apoptosis. Further studies are needed to determine whether either protein has diagnostic utility. [ABSTRACT FROM AUTHOR]

Objective--To characterize historical, clinicopathologic, ultrasonographic, microbiological, surgical, and histopathologic features of bacterial cholecystitis and bactibilia in dogs and evaluate response to treatment and outcomes in these patients. Design--Retrospective case-control study. Animals--40 client-owned dogs (10 with bacterial cholecystitis on histologic analysis or bactibilia on cytologic examination [case dogs] and 30 without bactibilia [controls]) evaluated at a veterinary teaching hospital between 2010 and 2014. Procedures--Signalment, history, clinicopathologic findings, ultrasonographic features, microbiological results, surgical findings, histopathologic changes, treatments, and outcomes of case dogs were derived from medical records and summarized. Demographic and clinicopathologic data and ultrasonographic findings were compared between case and control dogs. Relationships among prior antimicrobial treatment, sediment formation in the gallbladder, presence of immobile biliary sludge, and presence of bactibilia or bacterial cholecystitis were assessed. Results--No finding was pathognomonic for bactibilia or bacterial cholecystitis in dogs. Case dogs were significantly more likely to have immobile biliary sludge and had a greater degree of biliary sediment formation than did control dogs. All case dogs for which gallbladders were examined histologically (6/6) had bacterial cholecystitis. Five of 10 case dogs were Dachshunds. Medical or surgical treatment resulted in good outcomes. Conclusions and Clinical Relevance--Bactibilia and bacterial cholecystitis were important differential diagnoses in dogs with signs referable to biliary tract disease. Dachshunds were overrepresented, which may suggest a breed predisposition. Cytologic evaluation of bile should be considered in the routine assessment of dogs with hepatobiliary disease if immobile biliary sludge is present. [ABSTRACT FROM AUTHOR]

Background: Mounting evidence from human studies suggests that bile acid dysmetabolism might play a role in various human chronic gastrointestinal diseases. It is unknown whether fecal bile acid dysmetabolism occurs in dogs with chronic inflammatory enteropathy (CE). Objective: To assess microbial dysbiosis, fecal unconjugated bile acids (fUBA), and disease activity in dogs with steroid‐responsive CE. Animals: Twenty‐four healthy control dogs and 23 dogs with steroid‐responsive CE. Methods: In this retrospective study, fUBA were measured and analyzed. Fecal microbiota were assessed using a dysbiosis index. The canine inflammatory bowel disease activity index was used to evaluate remission of clinical signs. This was a multi‐institutional study where dogs with steroid‐responsive CE were evaluated over time. Results: The dysbiosis index was increased in dogs with CE (median, 2.5; range, −6.2 to 6.5) at baseline compared with healthy dogs (median, −4.5; range, −6.5 to −2.6; P = .002) but did not change in dogs with CE over time. Secondary fUBA were decreased in dogs with CE (median, 29%; range, 1%‐99%) compared with healthy dogs (median, 88%; 4%‐96%; P = .049). The percent of secondary fUBA in dogs with CE increased from baseline values (median, 28%; range, 1%‐99%) after 2‐3 months of treatment (median, 94%; range, 1%‐99%; P = 0.0183). Conclusions and Clinical Importance: These findings suggest that corticosteroids regulate fecal bile acids in dogs with CE. Additionally, resolution of clinical activity index in dogs with therapeutically managed CE and bile acid dysmetabolism are likely correlated. However, subclinical disease (i.e., microbial dysbiosis) can persist in dogs with steroid‐responsive CE. [ABSTRACT FROM AUTHOR]