Richard E. Goldstein

Adherent and invasive Escherichia coli is associated with granulomatous colitis in boxer dogs.

ABSTRACT The mucosa-associated microflora is increasingly considered to play a pivotal role in the pathogenesis of inflammatory bowel disease. This study explored the possibility that an abnormal mucosal flora is involved in the etiopathogenesis of granulomatous colitis of Boxer dogs (GCB). Colonic biopsy samples from affected dogs (n = 13) and controls (n = 38) were examined by fluorescent in situ hybridization (FISH) with a eubacterial 16S rRNA probe. Culture, 16S ribosomal DNA sequencing, and histochemistry were used to guide subsequent FISH. GCB-associated Escherichia coli isolates were evaluated for their ability to invade and persist in cultured epithelial cells and macrophages as well as for serotype, phylogenetic group, genome size, overall genotype, and presence of virulence genes. Intramucosal gram-negative coccobacilli were present in 100% of GCB samples but not controls. Invasive bacteria hybridized with FISH probes to E. coli. Three of four GCB-associated E. coli isolates adhered to, invaded, and replicated within cultured epithelial cells. Invasion triggered a“ splash”-type response, was decreased by cytochalasin D, genistein, colchicine, and wortmannin, and paralleled the behavior of the Crohns disease-associated strain E. coli LF 82. GCB E. coli and LF 82 were diverse in serotype and overall genotype but similar in phylogeny (B2 and D), in virulence gene profiles (fyuA, irp1, irp2, chuA, fepC, ibeA, kpsMII, iss), in having a larger genome size than commensal E. coli, and in the presence of novel multilocus sequence types. We conclude that GCB is associated with selective intramucosal colonization by E. coli. E. coli strains associated with GCB and Crohns disease have an adherent and invasive phenotype and novel multilocus sequence types and resemble E. coli associated with extraintestinal disease in phylogeny and virulence gene profile.

2010 ACVIM Small Animal Consensus Statement on Leptospirosis: Diagnosis, Epidemiology, Treatment, and Prevention

This report offers a consensus opinion on the diagnosis, epidemiology, treatment, and prevention of leptospirosis in dogs, an important zoonosis. Clinical signs of leptospirosis in dogs relate to development of renal disease, hepatic disease, uveitis, and pulmonary hemorrhage. Disease may follow periods of high rainfall, and can occur in dogs roaming in proximity to water sources, farm animals, or wildlife, or dogs residing in suburban environments. Diagnosis is based on acute and convalescent phase antibody titers by the microscopic agglutination test (MAT), with or without use of polymerase chain reaction assays. There is considerable interlaboratory variation in MAT results, and the MAT does not accurately predict the infecting serogroup. The recommended treatment for optimal clearance of the organism from renal tubules is doxycycline, 5 mg/kg PO q12h, for 14 days. Annual vaccination can prevent leptospirosis caused by serovars included in the vaccine and is recommended for dogs at risk of infection.

Clinicopathologic, histologic, and toxicologic findings in 70 cats inadvertently exposed to pet food contaminated with melamine and cyanuric acid

OBJECTIVE To document clinicopathologic, histologic, and toxicologic findings in cats inadvertently exposed to pet food contaminated with melamine and cyanuric acid. DESIGN Case series. ANIMALS 70 cats from a single cattery inadvertently fed contaminated food that was the subject of a March 2007 recall. PROCEDURES Clinical signs, clinicopathologic and histopathologic findings, and results of toxicologic analyses were recorded. RESULTS Clinical signs were identified in 43 cats and included inappetence, vomiting, polyuria, polydipsia, and lethargy. Azotemia was documented in 38 of the 68 cats for which serum biochemical analyses were performed 7 to 11 days after consumption of the contaminated food. One cat died, and 13 were euthanized. Histologic examination of kidney specimens from 13 cats revealed intratubular crystalluria, tubular necrosis with regeneration, and subcapsular perivascular inflammation characterized by perivascular fibroplasia or fibrosis and inflammation with intravascular fibrin thrombi. Toxicologic analyses revealed melamine and cyanuric acid in samples of cat food, vomitus, urine, and kidneys. CONCLUSIONS AND CLINICAL RELEVANCE In cats unintentionally fed pet food contaminated with melamine and cyanuric acid, the most consistent clinical and pathologic abnormalities were associated with the urinary tract, specifically tubular necrosis and crystalluria.

Urinary tract infections in cats with hyperthyroidism, diabetes mellitus and chronic kidney disease

The prevalence of urinary tract infections (UTIs) in cats with hyperthyroidism (n=90), diabetes mellitus (DM) (n=57) and chronic kidney disease (CKD) (n=77) was evaluated retrospectively. It was found to be 12% in cats with hyperthyroidism and DM, respectively, and 22% in cats with CKD. Associations between UTIs and clinical signs, biochemical markers in serum and urinalyses were investigated. Many of the cats with UTIs had no clinical signs of lower urinary tract disease or changes in their laboratory values indicative of infection. Therefore, a urinalysis alone should not be used to exclude UTIs in these cats. UTIs are relatively common in cats with hyperthyroidism, DM and CKD, and urine cultures are recommended as part of the basic diagnostic plan for cats suspected of suffering from these conditions.

Outcome of cats with low-grade lymphocytic lymphoma: 41 cases (1995-2005).

OBJECTIVE To evaluate factors associated with response to treatment, remission duration, and survival in cats with low-grade lymphoma affecting various organ systems. DESIGN Retrospective case series. SAMPLE POPULATION 41 cats with histologically confirmed low-grade lymphocytic lymphoma. PROCEDURES Medical records and biopsy specimens of cats with histologically confirmed low-grade lymphocytic lymphoma of various organ systems treated with prednisone and chlorambucil between 1995 and 2005 were reviewed. The Kaplan-Meier method was used to estimate remission duration and survival. Factors potentially associated with prognosis were compared. RESULTS Common clinical signs were weight loss (83%), vomiting (73%), anorexia (66%), and diarrhea (58%). Seventy-eight percent of cats tested had low serum cobalamin concentrations. Lymphoma was confined to the gastrointestinal tract in 68% of cats. Fifty-six percent of cats achieved a complete response to treatment, and 39% achieved a partial response. Five percent of cats had no response. No association was found between any risk factors (including anatomic site) and response to treatment. Partial response was associated with shorter remission duration, compared with complete response; median remission duration was 428 days for cats achieving a partial response, compared with 897 days for cats achieving a complete response. No other factors were associated with remission duration. Overall median survival time was 704 days. No factors were significantly associated with survival time. CONCLUSIONS AND CLINICAL RELEVANCE Most cats with lymphocytic lymphoma responded to treatment with prednisone and chlorambucil, and most factors evaluated were not associated with outcome.

Association of periodontal disease with systemic health indices in dogs and the systemic response to treatment of periodontal disease

OBJECTIVE To determine whether severity of periodontal disease (PD) was associated with systemic health indices in dogs and whether treatment of PD altered systemic health indices. DESIGN Prospective cohort study. ANIMALS 38 dogs. PROCEDURES Healthy dogs with clinical signs of PD were included in the study. Physical examination, serum biochemical analysis, a CBC, urine evaluation, measurement of serum C-reactive protein (CRP) concentration, and a microalbuminuria test were performed prior to treatment of PD. All tooth roots were scored for gingivitis and attachment loss, and appropriate treatment of PD was performed. Laboratory data were obtained 4 weeks after treatment. The Spearman rank correlation and Wilcoxon signed rank test were used for statistical analysis. RESULTS Analyses of the correlation of several variables with attachment loss or gingivitis or of differences before and after treatment revealed significant results for several variables. After applying Bonferroni corrections for family-wise error rate, significant rank correlations were found between attachment loss and platelet number (r = 0.54), creatinine concentration (r = -0.49), and the within-dog difference in CRP concentrations before and after treatment (r = 0.40). The BUN concentration was significantly higher after treatment than before treatment. CONCLUSIONS AND CLINICAL RELEVANCE Increasing severity of attachment loss was associated with changes in systemic inflammatory variables and renal indices. A decrease in CRP concentration after treatment was correlated with the severity of PD. The BUN concentration increased significantly after treatment of PD. There is a need for continued research into the systemic impact of PD.

Dirofilaria repens infection in a dog: diagnosis and treatment with melarsomine and doramectin

Therapy of canine dirofilariois due to Dirofilaria repens is indicated for dogs suffering from clinical signs of this disease, such as dermal swelling, sub-cutaneous nodules and pruritus. It is also important in order to decrease the risk of infection to other dogs and humans in the vicinity of the infected animal when suitable mosquito vectors are present. Combined therapy with the arsenic adulticide melarsomine and the avermectin microfilaricidal doramectin was effective in clearing infection with D. repens in a dog. The number of microfilariae dropped from 17 microl(-1) blood pre-treatment to 7 microl(-1) following the first adulticide injection and reached 0 a day after the microfilaricidal administration. The dog remained negative for D. repens microfilaremia during a follow-up period of 90 days. Euthanasia and necropsy performed 3 months after the initiation of therapy due to a progressive neoplastic disease revealed no evidence of filariae.

Effects of Recent Leptospira Vaccination on Whole Blood Real-Time PCR Testing in Healthy Client-Owned Dogs

BACKGROUND Bacterin-based canine Leptospira vaccines could present a challenge for the use of whole blood real-time polymerase chain reaction (PCR) as a diagnostic tool. Recent vaccination could induce positive results if the targeted DNA fragment is present within the vaccine and in the blood of the recently vaccinated dog. OBJECTIVES The objective of this study was to assess whether 2 available 4-serovar vaccines induce a positive real-time PCR reaction in the blood of healthy recently vaccinated dogs. ANIMALS Twenty healthy dogs. METHODS This was a prospective study. Dogs were assigned to 1 of 2 vaccine groups. Both vaccines were culture-based and include Leptospira interrogans serovars Pomona, Canicola, and Icterohaemorrhagiae and Leptospira kirschneri serovar Grippotyphosa. Whole blood for real-time PCR and serum for the microscopic agglutination test (MAT) were collected prior to and 3 and 7 days after vaccination and weekly thereafter for 8 weeks. Two real-time PCR tests targeting 2 different genes were performed independently in a blinded fashion. RESULTS Both Leptospira vaccines produced positive real-time PCR reactions when assayed undiluted or diluted 1 : 100 in canine blood. However, blood samples drawn from all dogs at all time points after vaccination were negative on PCR. All dogs developed MAT titers. CONCLUSIONS AND CLINICAL IMPORTANCE Recent vaccination with 2 commercially available vaccines does not interfere with the use of real-time PCR for the identification of acute Leptospira infection in dogs.

Microalbuminuria and Comparison of Serologic Testing for Exposure to Borrelia Burgdorferi in Nonclinical Labrador and Golden Retrievers

Canine Lyme disease is caused by the spirochete Borrelia burgdorferi after transmission by an Ixodes tick, typically resulting in joint pain, fever and lethargy. Lyme nephritis is a poorly characterized syndrome associated with severe glomerular and tubular renal injury and poor clinical outcome in young to middle-aged dogs positive for exposure to B. burgdorferi. The aims of this study were to identify associations between natural exposure to B. burgdorferi and the presence of microalbuminuria in nonclinical young Labrador and Golden Retrievers and to compare two commonly used serologic tests available to document B. burgdorferi exposure: the Western blot and the commercial point-of-care C6 peptide enzyme-linked immunosorbent assay (ELISA) tests. Microalbuminuria was assessed using a commercial point-of-care ELISA specific for canine albumin. Blood and urine samples from 268 asymptomatic Labrador and Golden Retrievers were included. Of these, 18.7% were positive for B. burgdorferi exposure according to the C6 ELISA; 21.2% were positive for natural exposure to B. burgdorferi and 11.5% for vaccinal antibodies according to the Western blot. The agreement rate was 93% between the two tests (kappa = 0.78, P < 0.0001) for natural exposure. Urine from 6.1% of the dogs was positive for microalbuminuria. There was no association between microalbuminuria and exposure to B. burgdorferi based on results of a Western blot (P = 0.57) or C6 ELISA (P = 0.53). Microalbuminuria is likely not a consequence of B. burgdorferi exposure in young nonclinical Labrador and Golden Retrievers.

Single-dose pharmacokinetics and genotoxicity of metronidazole in cats.

Single-dose pharmacokinetics and genotoxicity of metronidazole in cats were evaluated. Cats received either 5 mg/kg metronidazole intravenously, or 20 mg/kg metronidazole benzoate (12.4 mg/kg metronidazole base) orally in a single dose. Serial plasma samples were collected and assayed for metronidazole using high pressure liquid chromatography (HPLC). Genotoxicity was assessed in vitro in feline peripheral blood mononuclear cells (PBMC) and a feline T-cell lymphoma line incubated with metronidazole, and in vivo in PBMC collected before, during and 7 days after oral metronidazole, by use of the COMET assay. Systemic absorption of metronidazole was variable (mean=65±28%) with a peak of 8.84±5.4 μg/ml at 3.6±2.9 h. The terminal half-life was 5.34 h from the intravenous dose and 5.16 h from the oral dose. Systemic clearance was low (mean=91.57 ml/h/kg [1.53 ml/kg/min]), and the apparent volume of distribution (steady state) was 0.650±0.254 l/kg. Genotoxicity was detected at all concentrations of metronidazole in feline PBMC and the T-cell lymphoma line in vitro. Genotoxicity was also observed in PBMC collected from cats after 7 days of oral metronidazole but resolved within 6 days of discontinuing metronidazole.