Untargeted metabolomics reveals the effects of pre-analytic storage on serum metabolite profiles from healthy cats.

Untargeted metabolomics investigations have characterized metabolic disturbances associated with various diseases in domestic cats. However, the pre-analytic stability of serum metabolites in the species is unknown. Our objective was to compare serum metabolomes from healthy cats stored at -20°C for up to 12 months to samples stored at -80°C. Serum samples from 8 adult, healthy cats were stored at -20°C for 6 months, -20°C for 12 months, or -80°C for 12 months. Untargeted liquid chromatography-mass spectrometry was used to generate serum metabolite profiles containing relative abundances of 733 serum metabolites that were compared among storage conditions. Unsupervised analysis with principal component analysis and hierarchical clustering of Euclidian distances revealed separation of samples from individual cats regardless of storage condition. Linear mixed-effects models identified 75 metabolites that differed significantly among storage conditions. Intraclass correlation analysis (ICC) classified most serum metabolites as having excellent (ICC ≥ 0.9; 33%) or moderate (ICC 0.75-0.89; 33%) stability, whereas 13% had poor stability (ICC < 0.5). Biochemicals that varied significantly among storage conditions and classified with poor stability included glutathione metabolites, amino acids, gamma-glutamyl amino acids, and polyunsaturated fatty acids. The benzoate; glycine, serine and threonine; tryptophan; chemical (xenobiotics); acetylated peptide, and primary bile acid sub pathways were enriched among highly stable metabolites, whereas the monohydroxy fatty acid, polyunsaturated fatty, and monoacylglycerol sub-pathways were enriched among unstable metabolites. Our findings suggest that serum metabolome profiles are representative of the cat of origin, regardless of storage condition. However, changes in specific serum metabolites, especially glutathione, gamma-glutamyl amino acid, and fatty acid metabolites were consistent with increased sample oxidation during storage at -20°C compared with -80°C. By investigating the pre-analytic stability of serum metabolites, this investigation provides valuable insights that could aid other investigators in planning and interpreting studies of serum metabolomes in cats.

Untargeted Analysis of Serum Metabolomes in Dogs with Exocrine Pancreatic Insufficiency.

Exocrine pancreatic insufficiency (EPI) is a malabsorptive syndrome resulting from insufficient secretion of pancreatic digestive enzymes. EPI is treated with pancreatic enzyme replacement therapy (PERT), but the persistence of clinical signs, especially diarrhea, is common after treatment. We used untargeted metabolomics of serum to identify metabolic disturbances associated with EPI and generate novel hypotheses related to its pathophysiology. Fasted serum samples were collected from dogs with EPI (n = 20) and healthy controls (n = 10), all receiving PERT. Serum metabolomes were generated using UPLC-MS/MS, and differences in relative metabolite abundances were compared between the groups. Of the 759 serum metabolites detected, 114 varied significantly (p < 0.05, q < 0.2) between dogs with EPI and healthy controls. Differences in amino acids (arginate, homoarginine, 2-oxoarginine, N-acetyl-cadaverine, and α-ketoglutaramate) and lipids (free fatty acids and docosahexaenoylcarnitine) were consistent with increased proteolysis and lipolysis, indicating a persistent catabolic state in dogs with EPI. Relative abundances of gut microbial metabolites (phenyllactate, 4-hydroxyphenylacetate, phenylacetyl-amino acids, catechol sulfates, and o-cresol-sulfate) were altered in dogs with EPI, consistent with disruptions in gut microbial communities. Increased kynurenine is consistent with the presence of intestinal inflammation in dogs with EPI. Whether these metabolic disturbances participate in the pathophysiology of EPI or contribute to the persistence of clinical signs after treatment is unknown, but they are targets for future investigations.

Association between Mycoplasma-specific polymerase chain reaction assay results and oral bacterial contamination of bronchoalveolar lavage fluid samples from dogs with respiratory tract disease: 121 cases (2005-2012).

OBJECTIVE: To determine whether an association exists between oral bacterial contamination of bronchoalveolar lavage fluid (BALF) and positive PCR assay results for the detection of Mycoplasma spp in BALF samples of dogs with lower respiratory tract (LRT; portion from the trachea to the lungs) disease. DESIGN: Retrospective case series. ANIMALS: 121 dogs with LRT disease. Procedures-Medical records from January 2005 to April 2012 were reviewed. Dogs with LRT disease that had BALF samples evaluated by use of Mycoplasma-specific PCR assay, bacterial culture, and cytologic examination were included. Information on signalment, final diagnoses, and BALF testing results was extracted. RESULTS: 83 (68.6%) dogs had BALF samples with negative PCR assay results for Mycoplasma spp, and 38 (31.4%) had positive results. The BALF samples with cytologic evidence of oral bacterial contamination were 5.1 times as likely to have positive Mycoplasma-specific PCR assay results as were noncontaminated samples. Compared with hound or herding dogs, other breeds were 13.6 times as likely to have positive PCR assay results. Dogs with bronchitis were less likely than dogs with other LRT diseases to have positive Mycoplasma-specific PCR assay results. No significant association was found between Mycoplasma-specific PCR assay results and bacterial culture results. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs with LRT disease, Mycoplasma-specific PCR assay results for BALF samples should be interpreted in terms of possible oral bacterial contamination. Mycoplasma-specific PCR assay of BALF samples from herding dogs, hound dogs, and dogs with bronchitis may be less rewarding than for other dogs with LRT disease.

Thyrotoxicosis in a dog induced by the consumption of feces from a levothyroxine-supplemented housemate.

A 9-year-old golden retriever dog was evaluated for polyuria, polydipsia, weight loss, and elevated serum thyroxine. Targeted questioning revealed that the dog was coprophagic and routinely ingested the feces of a dog that was treated with twice-daily levothyroxine. Clinical signs resolved and serum thyroxine decreased to normal levels in the affected dog with prevention of coprophagy.

Thyrotoxicose chez un chien induite par la consommation de fèces d'un compagnon prenant des suppléments de lévothyroxine. Un chien Golden retriever âgé de 9 ans a été évalué pour la polyurie, la polydipsie, la perte de poids et la thyroxine sérique élevée. Des questions ciblées ont révélé que le chien était coprophage et ingérait régulièrement les fèces d'un chien qui était traité deux fois par jour à la lévothyroxine. Les signes cliniques se sont résorbés et la thyroxine sérique a baissé à des niveaux normaux chez le chien affecté avec la prévention de la coprophagie.(Traduit par Isabelle Vallières).

Coercive control and abused women's decisions about their pets when seeking shelter.

The importance of pets in families, especially during major life stressors, is well documented. Research suggests links between pet ownership and intimate partner violence (IPV). This study explored abused women's decisions about pets when seeking help from a shelter. Interviews were conducted with 19 women who were pet owners. Using grounded theory methods, two patterns emerged surrounding abusers' treatment of pets, bonds to pets, women's decisions about pets upon seeking shelter, and future plans for pets. The presence of coercive control was central to these patterns. Women also discussed their experiences with and needs from shelter professionals and veterinarians with implications for practice.

Incidence of bacteremia following upper gastrointestinal endoscopy and biopsy in healthy dogs before, during, and after treatment with omeprazole.

OBJECTIVE: To determine the incidence of bacteremia, as detected by routine methods for bacterial culture of blood samples, following routine endoscopic biopsy of the stomach and duodenum in healthy research dogs and to determine whether treatment with omeprazole administration affected the incidence of bacteremia. ANIMALS: 8 healthy purpose-bred research dogs. PROCEDURES: All dogs underwent gastroduodenoscopy with biopsy at 4 points: twice prior to treatment with omeprazole, once following 15 days of omeprazole treatment (20 mg, PO, q 12 h), and once 14 days after treatment ceased. Dogs had a mean ± SD body weight of 18.6 ± 2.0 kg. Blood samples were aseptically obtained at 3 points during each procedure (before, immediately following, and 24 hours after endoscopy), and routine aerobic and anaerobic bacterial culture of blood was performed. RESULTS: 96 cultures were attempted for each culture method, yielding positive results of aerobic culture for 2 dogs at separate time points and no positive results of anaerobic culture. CONCLUSIONS AND CLINICAL RELEVANCE: Routine gastrointestinal endoscopy with biopsy in healthy dogs did not result in a detectable bacteremia in most dogs. Treatment with the gastric acid-suppressing medication omeprazole did not affect the incidence of bacteremia as detected via standard techniques.

An unusual clinical presentation of a dog with gastrinoma.

Gastrinoma is a rare malignant neuroendocrine neoplasia that results in autonomous gastrin secretion that stimulates hypersecretion of gastric acid, resulting in severe gastric and proximal small intestinal ulcerations. The principal clinical manifestation of gastrinoma is persistent vomiting. This report describes an uncommon manifestation of pancreatic gastrinoma in a dog.

Blastomyces dermatitidis prostatic and testicular infection in eight dogs (1992-2005).

This was a retrospective case study of eight dogs diagnosed with prostatic or testicular B. dermatitidis infection. Signalment, clinical presentation, diagnostic procedures, and treatment options were evaluated. Review of medical records of dogs diagnosed with blastomycosis at the University of Illinois Veterinary Teaching Hospital from 1992 to 2005 yielded four dogs with prostatic blastomycosis (PB) and four dogs with testicular blastomycosis (TB). Three of the four dogs with PB and all four dogs with TB had evidence of urogenital disease. Three dogs with PB had an elevated body temperature and all had systemic disease. All dogs with TB had a normal body temperature, and three had systemic disease and one had clinical signs limited to testicular disease. Cytology or histopathology was used to diagnose PB or TB. Treatment included itraconazole or fluconazole with or without nonsteroidal anti-inflammatory drugs. PB and TB are infrequently recognized and may be under diagnosed due to failure to specifically evaluate these tissues. PB or TB should be considered in the evaluation and staging of male dogs with blastomycosis. Male dogs with urogenital signs should be evaluated via prostatic or testicular cytology or histopathology since proper identification and management of PB or TB may improve overall treatment success.

DIC and granulomatous vasculitis in a dog with disseminated histoplasmosis.

Disseminated histoplasmosis and disseminated intravascular coagulopathy were diagnosed in a 7 mo old, female spayed mixed-breed dog. The dog improved transiently with supportive care, but deteriorated shortly after initiation of antifungal therapy. The dog was subsequently euthanized. At necropsy, marked granulomatous vasculitis was identified in all affected organs. The tunicae and laminae of the arteries and arterioles were obscured by epithelioid macrophages and multinucleated giant cells admixed with necrotic material. Intracytoplasmic yeast were present within some of these macrophages. To the authors' knowledge, this is the first reported case of granulomatous vasculitis associated with Histoplasma capsulatum in a dog.

Detection of occult urinary tract infections in dogs with diabetes mellitus.

Dogs with diabetes mellitus may develop occult urinary tract infections. In this study, diabetic dogs with negative and positive bacterial urine cultures were compared. Records from 51 dogs with diabetes mellitus were reviewed at the University of Illinois. No difference was identified between the groups in urine specific gravity, pH, glucose, ketones, protein, red blood cells, white blood cells, or epithelial cells. Dogs with occult urinary tract infection did have an increased incidence of bacteriuria, but this was not a consistent finding. Therefore, the urine on all diabetic dogs should be cultured to accurately identify the presence or absence of bacterial urinary tract infections.