Jacqueline C. Whittemore

Bioequivalence of Orally Administered Generic, Compounded, and Innovator‐Formulated Itraconazole in Healthy Dogs

Background Itraconazole is commonly used to treat systemic fungal infections in dogs, but problems exist with absorption and cost. Objective To determine oral bioequivalence of generic and compounded itraconazole compared to original innovator (brand name) itraconazole in healthy dogs. Animals Nine healthy, adult research Beagle dogs. Methods A randomized, 3‐way, 3‐period, crossover design with an 8‐day washout period. After a 12‐hour fast, each dog received 100 mg (average: 10.5 mg/kg) of either innovator itraconazole, an approved human generic capsule, or compounded itraconazole (compounded using a commercially available compounding vehicle) with a small meal. Plasma was collected at predetermined intervals for high pressure liquid chromatography analysis. Concentration data were analyzed using noncompartmental pharmacokinetics to determine area under the curve (AUC), peak concentration (CMAX), and terminal half‐life. Bioequivalence tests compared generic and compounded itraconazole to the reference formulation. Results Average ratios of compounded and generic formulations to the reference formulation of itraconazole for AUC were 5.52% and 104.2%, respectively, and for CMAX were 4.14% and 86.34%, respectively. A test of bioequivalence using 2 one‐sided tests and 90% confidence intervals did not meet bioequivalence criteria for either formulation. Conclusion and Clinical Importance Neither generic nor compounded itraconazole is bioequivalent to the reference formulation in dogs. However, pharmacokinetic data for generic formulation were similar enough that therapeutic concentrations could be achieved. Compounded itraconazole produced such low plasma concentrations, it is unlikely to be effective; therefore, compounded itraconazole should not be used in dogs.

Comparison of Signalment, Clinicopathologic Findings, Histologic Diagnosis, and Prognosis in Dogs with Glomerular Disease with or without Nephrotic Syndrome

BACKGROUND Nephrotic syndrome (NS) develops most commonly in people with glomerular diseases associated with marked albuminuria. Hypernatremia, hypertension, and progressive renal failure are more prevalent in nephrotic than nonnephrotic human patients. HYPOTHESIS/OBJECTIVES Dogs with NS have higher serum cholesterol, triglyceride, and sodium concentrations, higher urine protein:creatinine ratios (UPC) and systolic blood pressure, and lower serum albumin concentrations than dogs with nonnephrotic glomerular disease (NNGD). NS is associated with membranous glomerulopathy and amyloidosis. Affected dogs are more likely to be azotemic and have shorter survival times. ANIMALS Two hundred and thirty-four pet dogs (78 NS dogs, 156 NNGD dogs). METHODS Multicenter retrospective case-control study comparing time-matched NS and NNGD dogs. NS was defined as the concurrent presence of hypoalbuminemia, hypercholesterolemia, proteinuria, and extravascular fluid accumulation. Signalment, clinicopathologic variables, histopathologic diagnoses, and survival time were compared between groups. RESULTS Age, serum albumin, chloride, calcium, phosphate, creatinine, and cholesterol concentrations, and UPC differed significantly between NS and NNGD dogs. Both groups were equally likely to be azotemic at time of diagnosis, and NS was not associated with histologic diagnosis. Median survival was significantly shorter for NS (12.5 days) versus NNGD dogs (104.5 days). When subgrouped based on serum creatinine (< or ≥1.5 mg/dL), survival of NS versus NNGD dogs was only significantly different in nonazotemic dogs (51 versus 605 days, respectively). CONCLUSIONS AND CLINICAL IMPORTANCE Presence of NS is associated with poorer prognosis in dogs with nonazotemic glomerular disease. Preventing development of NS is warranted; however, specific interventions were not evaluated in this study.

Associations among albuminuria, C-reactive protein concentrations, survival predictor index scores, and survival in 78 critically ill dogs.

BACKGROUND Microalbuminuria and C-reactive protein (CRP) are predictors of morbidity and survival in critically ill human patients. HYPOTHESIS/OBJECTIVES To evaluate results of microalbuminuria assays (untimed single-sample urine albumin concentration [U-ALB] and the urine albumin:creatinine ratio [UACR]), serum CRP, and survival predictor index (SPI2) scores as predictors of survival in critically ill dogs. ANIMALS Seventy-eight dogs admitted to intensive care units at University of Tennessee (UT) and Colorado State University (CSU). METHODS Prospective observational study. Critically ill dogs were eligible for enrollment, unless euthanized because of financial constraints. Samples were collected within 3 hours of admission. Spearmans rank-correlation coefficients were determined for U-ALB, UACR, CRP, and SPI2. U-ALB, UACR, CRP, and SPI2 were assessed for associations with 7- and 30-day survival by Mann-Whitney U-tests and receiver operating characteristic (ROC) curves. P-values < .0125 were considered significant. RESULTS UT (n = 49) and CSU (n = 29) patients did not differ significantly. Forty percent (31/78) of dogs died. SPI2 was inversely correlated with U-ALB (r(s) = -0.39, P < .001) and UACR (r(s) = -0.41, P < .001). CRP was not correlated with SPI2 (P = .019), U-ALB (P > .1), or UACR (P > .1). U-ALB and UACR had very high correlation (r(s) = 0.95, P < .001). SPI2, U-ALB, and UACR differed significantly for survivors and nonsurvivors. SPI2, U-ALB, and UACR had areas under the ROC curve (AUC) from 0.68 to 0.74 for survival prediction. CONCLUSIONS AND CLINICAL IMPORTANCE Albuminuria and SPI2, but not CRP, are associated with survival in critically ill dogs. Suboptimal AUCs limit the value of microalbuminuria testing for clinical risk assessment. Additional studies are necessary to determine the usefulness of microalbuminuria testing in patient risk stratification for prospective research.

Antibodies against Crandell Rees Feline Kidney (CRFK) Cell Line Antigens, α‐Enolase, and Annexin A2 in Vaccinated and CRFK Hyperinoculated Cats

BACKGROUND Cats inoculated with feline herpesvirus 1, calicivirus, and panleukopenia (FVRCP) vaccines grown on the Crandell Rees feline kidney (CRFK) cell line have been shown to develop anti-CRFK antibodies. The identities of common CRFK antigens are unknown. HYPOTHESIS Cats inoculated with CRFK lysates and FVRCP vaccines will develop autoantibodies measurable by Western blot immunoassay. Antigens associated with these antibodies can be isolated for further study. ANIMALS One CRFK hyperinoculated rabbit, 44 age-matched unvaccinated kittens purchased from a commercial vendor. METHODS Commonly recognized CRFK antigens were identified by comparison of Western blot immunoassays using sera from a hyperinoculated rabbit and kittens inoculated with CRFK lysate or 1 of 4 commercially available FVRCP vaccines. Antigens were purified from CRFK lysates and sequenced. Antigen recognition was confirmed by Western blot immunoassay and indirect ELISA for 2 proteins using sera from CRFK and FVRCP inoculated kittens. RESULTS CRFK antigens 47, 40, and 38 kD in size were identified. Protein isolation and sequencing identified 3 CRFK proteins as alpha-enolase, annexin A2, and macrophage capping protein (MCP). Sera from FVRCP and CRFK inoculated cats were confirmed to recognize annexin A2 and alpha-enolase by Western blot immunoassay and indirect ELISA. CONCLUSIONS AND CLINICAL RELEVANCE This study validated the use of Western blot immunoassay for detection of antibodies against CRFK proteins and identified 3 CRFK antigens. In humans, alpha-enolase antibodies are nephritogenic; alpha-enolase and annexin A2 antibodies have been associated with autoimmune diseases. Further research will be necessary to determine the clinical relevance of these findings.

Canine Pancreatic-Specific Lipase Concentrations in Clinically Healthy Dogs and Dogs with Naturally Occurring Hyperadrenocorticism

Background Specificity of canine pancreatic lipase immunoreactivity (cPLI) assays in dogs with hyperadrenocorticism (HAC) is unknown. Hypothesis Results of cPLI assays differ for clinically healthy dogs and dogs with HAC. Animals Seventeen healthy dogs and 20 dogs with HAC diagnosed by ACTH stimulation test results without evidence of clinical pancreatitis. Methods Dogs were enrolled between December 2009 and November 2010. Serum cPLI concentrations were determined by quantitative (Spec cPL test, SPEC) and semiquantitative (SNAP cPL test, SNAP) assays. Results were categorized as normal, equivocal, or abnormal (SPEC) or negative or positive (SNAP). Associations between group and cPLI were assessed using Fishers exact test or the Mann–Whitney U‐test. Spearman rank correlation coefficients (ρ) were determined for SNAP and SPEC results. Significance was set at P < .05. Results Spec cPL test concentrations were significantly (P < .001) higher in dogs with HAC (491.1 μg/L) than in healthy dogs (75.2 μg/L), with more abnormal SPEC results in HAC dogs (P < .001). There were more (P = .002) positive SNAP results in dogs with HAC (55%) than in healthy dogs (6%). SNAP and SPEC results were highly correlated (ρ = 0.85; P < .001). Conclusions and Clinical Importance Dogs with HAC had higher SPEC concentrations and more positive SNAP results than clinically healthy dogs with normal ACTH stimulation test results. Specificity of SPEC and SNAP assays in HAC dogs without clinical pancreatitis were 65 and 45%, respectively. Pending further study, SNAP and SPEC results should be interpreted cautiously in dogs with HAC to avoid false diagnosis of concurrent pancreatitis.

LAPAROSCOPIC-ASSISTED OVARIECTOMY OF TIGERS (PANTHERA TIGRIS) WITH THE USE OF THE LIGASURE™ DEVICE

Laparoscopic ovariectomy was performed in seven tigers with the use of a vessel-sealing device and a three-port technique. A comparison group of seven tigers that underwent traditional ovariohysterectomy was assembled with the use of a medical records search. Mean operative times for laparoscopic ovariectomy were compared to standard ovariohysterectomy, and mean combined laparoscopic incision length compared to standard ovariohysterectomy incision lengths. Significance was set at P < or = 0.05. Mean surgical time for laparoscopic ovariectomy (82 min, range 71-126 min) was significantly shorter than standard ovariohysterectomy surgical time (129 min, range 80-165 min, P = 0.007). Mean combined laparoscopic incision length (8.07 cm, range 3.80-9.50 cm) was significantly shorter than the mean incision length for standard ovariohysterectomy (13.57 cm, range 12.00-20.00 cm, P = 0.009). There were no clinically important complications observed in either group. Laparoscopic ovariectomy has a significantly shorter surgical time and combined incision length compared to standard ovariohysterectomy in tigers, and appears to be a safe and rapid sterilization method for tigers. Equipment cost and the necessity for advanced training may limit its use in some institutions. Further prospective evaluation is warranted to determine whether it is associated with decreased morbidity, mortality, or cost.

Bartonella species antibodies and hyperglobulinemia in privately owned cats.

BACKGROUND Bartonella species are zoonotic agents and primary pathogens in cats. Hyperglobulinemia has been associated with bartonellosis in humans and cats. HYPOTHESIS/OBJECTIVES To evaluate for associations between Bartonella species immunoglobulin G (IgG) antibodies and serum biochemistry panel results in privately owned cats. ANIMALS 1,477 privately owned cats. METHODS Residual sera were collected after biochemical evaluation for this prospective, cross-sectional serosurvey. Bartonella species IgG ELISA was performed with a cutoff value of ≥ 1 : 64. Stepwise logistic regression analysis was performed with the endpoint titer as the outcome variable. The final statistical model included age, albumin, ALP activity, ALT activity, bilirubin, creatinine, glucose, and globulin as covariates. Serum protein electrophoresis was performed with serum from 50 cats with and without antibodies to Bartonella species and hyperglobulinemia. Sera from cats seropositive to Bartonella species and with hyperglobulinemia were assessed for evidence of exposure to other infectious agents associated with hyperglobulinemia. RESULTS Risk of seropositivity to Bartonella species was positively associated with the natural log of globulin concentration (OR = 11.90, 95% CI 6.15-23.02, P < .0001), and inversely associated with the natural log of glucose concentration (OR = 0.66, 95% CI 0.50-0.87, P = .004). Another explanation for hyperglobulinemia was not identified for most cats with Bartonella species antibodies. Hyperglobulinemia was primarily caused by polyclonal gammopathy in cats that were seronegative and seropositive for Bartonella species. CONCLUSIONS AND CLINICAL IMPORTANCE Hyperglobulinemia was significantly associated with seropositivity to Bartonella species. Testing for bartonellosis is warranted in cats with unexplained hyperglobulinemia and clinical or laboratory findings suggestive of bartonellosis.

Diagnostic Accuracy of Tissue Impedance Measurement Interpretation for Correct Veress Needle Placement in Canine Cadavers

OBJECTIVE To establish the diagnostic accuracy of tissue impedance measurement interpretation (TIMI) for determining correct versus incorrect Veress needle placement. STUDY DESIGN Prospective, randomized, blinded trial. STUDY POPULATION Canine cadavers (n = 20). METHODS Two laparoscopists (1 experienced, 1 novice), blinded to tissue impedance measurement results, placed reusable Veress needles in canine cadavers in a randomized order. A 3rd individual interpreted impedance measurements as consistent with correct or incorrect Veress needle placement. Veress needle tip locations were marked by injecting contrasting colors of India ink. Tissue dissection was performed to localize ink. Sensitivity, specificity, accuracy, precision, and kappa statistics for TIMI for placements by the experienced and novice laparoscopist were determined. P < .05 was considered significant. RESULTS TIMI identified 29/33 correct and 7/7 incorrect placements, respectively. Impedance measurement interpretation had 87.9% sensitivity, 100% specificity, 90% accuracy, and 100% precision overall. Agreement between TIMI and Veress needle location was moderate (kappa 0.50, P = .01) for placements by the experienced laparoscopist and very high (kappa 0.88, P < .01) for the novice laparoscopist. CONCLUSIONS Diagnostic accuracy of TIMI for Veress needle placement was higher than has been reported for other tests, and TIMI had a shallow learning curve. TIMI successfully detected all incorrect Veress needle placements. Further prospective evaluation of Veress needle placement with and without TIMI is warranted to determine if its use increases operator detection of inappropriate Veress needle placements or decreases installment phase complication rates.

Actinomyces endogenous endophthalmitis in a cat following multiple dental extractions

An 8-year-old, brachycephalic, mixed breed cat underwent full mouth tooth extractions for the treatment of tooth root abscessation. Subsequently, the cat developed anterior uveitis refractory to topical therapy that eventually necessitated enucleation. Actinomyces species were isolated from both the tooth root abscesses and the anterior chamber after enucleation. Histopathology of the enucleated eye revealed panophthalmitis with abundant intralesional bacteria morphologically consistent with Actinomyces. Between the time of tooth root extraction and enucleation (20 weeks), the cat was diagnosed with hyperthyroidism and treated with oral steroids for inflammatory bowel syndrome. We believe this report represents a rare case of endogenous endophthalmitis secondary to dental disease, possibly precipitated by concurrent immunosuppression.

Effects of various factors on Doppler ultrasonographic measurements of radial and coccygeal arterial blood pressure in privately owned, conscious cats

OBJECTIVE To assess the effects of age, body condition score (BCS), and muscle condition score (MCS) on radial and coccygeal systolic arterial blood pressure (SAP) in cats. DESIGN Prospective randomized trial. ANIMALS 66 privately owned cats enrolled between May and December 2010. PROCEDURES BCS and MCS of cats were assessed by 2 investigators; SAP was measured via Doppler ultrasonic flow detector, with cats positioned in right lateral or sternal recumbency for measurements at the radial or coccygeal artery, respectively, with order of site randomized. Associations among variables were assessed through correlation coefficients, partial correlation coefficients, and ANCOVA. RESULTS Interrater reliability for BCS and MCS assessment was high (correlation coefficients, 0.95 and 0.83, respectively). No significant effect was identified for order of SAP measurement sites. Coccygeal and radial SAP were positively correlated (ρ = 0.45). The median difference in coccygeal versus radial SAP was 19 mm Hg, but differences were not consistently positive or negative. Radial SAP was positively correlated with age (ρ = 0.48) and negatively correlated with MCS (ρ = -0.30). On the basis of the correlation analysis, the association between radial SAP and MCS reflected the confounding influence of age. Coccygeal SAP was not significantly correlated with age, BCS, or MCS. CONCLUSIONS AND CLINICAL RELEVANCE Use of the coccygeal artery is recommended to reduce the confounding effects of age and sarcopenia on Doppler ultrasonographic SAP measurements in cats. Additionally, monitoring for changes in MCS is recommended for cats undergoing serial SAP measurement.