Dermatitis associated with microfilariae (Filarioidea) in 10 dogs

Dermatitis associated with microfilariae of a nematode of the superfamily Filarioidea was diagnosed in 10 dogs from the western United States. Clinically, lesions were single or multiple papules and plaques with alopecia, scarring, erythema, ulceration, or crusting. Eight dogs had lesional pruritus. Microscopically, there was perivascular, periglandular, to interstitial inflammation, with many eosinophils and/or plasma cells, and scarring affecting the dermis and subcutis. Microgranulomas containing microfilaria were seen in six dogs. Microfilariae were noted in microgranulomas or free in the dermis or subcutis, but not in vessels. In one case, an adult female nematode emerged from a biopsy sample that was placed in physiologic saline. Study of this nematode revealed that it was a filarioid of the family Onchocercidae; it was identified as Acanthocheilonema sp. (syn: Dipetalonema ). Antigen tests of five dogs were negative for Dirofilaria immitis . The Knott tests and/or filter tests of nine dogs were negative for microfilariae. An indirect fluorescence antibody test of one dog was also negative for D. immitis . One dog was not evaluated for microfilariae.     

Quantification of normetanephrine in canine urine using ELISA: evaluation of factors affecting results

Catecholamine release increases in dogs with pheochromocytomas and in situations of stress. Although plasma catecholamines degrade rapidly, their metabolites, normetanephrine (NME) and metanephrine (ME), are stable in acidified urine. Our aim was to verify a human urine ELISA kit for the quantification of NME and ME in canine urine and to determine the effects on metabolite stability of sampling time (morning or midday) and day (ordinary or day spent in a clinic). We analyzed 179 urine samples from 17 healthy dogs. For NME, the mean intra-assay CV was 6.0% for all samples and 4.3% for the canine control; inter-assay CVs were 3.3, 3.8, and 12% for high and low concentration human urine positive controls supplied in the ELISA kit and a positive canine control, respectively; spike-recovery was 90–101%. For ME, mean intra-assay CV was 6.5% for samples and 9.0% for the canine control; inter-assay CVs were 12.7, 7.2, and 22.5% for high and low concentration human urine positive controls supplied in the ELISA kit and a positive canine control, respectively; spike-recovery was 85–89%. Dilution recovery was unsatisfactory for both metabolites. Based on our verification results, NME was selected for remaining analyses. We found no effect on NME concentrations of acidification or room temperature storage for up to 24 h. The NME:creatinine ratio was higher after the first of 3 clinic days compared to the same morning (111.2 ± 5.5 vs. 82.9 ± 5.3; p < 0.0001), but not on the other days. NME verification results were generally superior to ME. Dilution studies were unsatisfactory for both metabolites. Given that NME was stable without acidification at room temperature, urine samples can be collected at home. The clinic environment can cause higher NME:creatinine ratios, especially in unaccustomed dogs.     

Comparison of glucose concentrations in canine whole blood,plasma, and serum measured with a veterinary point-of-care glucometer

Previous studies have determined that, compared to whole blood, serum or plasma used in a portable blood glucometer (PBG) may provide more accurate results. We investigated the accuracy of a veterinary PBG (AlphaTRAK 2; Zoetis) for the measurement of glucose concentrations in serum, plasma, and whole blood compared to plasma glucose concentration measured by a biochemical analyzer. Blood samples from 53 client-owned dogs were collected. Lin concordance correlation coefficient (ρ_c) and Bland–Altman plots were used to determine correlation and agreement between the results obtained for the different sample types. Glucose concentration in whole blood measured by the veterinary PBG was more strongly correlated with the glucose concentration measured by the biochemical analyzer (ρ_c = 0.92) compared to plasma and serum glucose concentrations (ρ_c = 0.59 and 0.57, respectively). The mean differences between the glucose concentrations in whole blood, plasma, and serum measured by the veterinary PBG and the glucose concentration determined by the biochemical analyzer were 1.0, 6.3, and 6.7 mmol/L (18, 113, and 121 mg/dL), respectively. Our findings suggest that, when using this veterinary PBG, the accuracy of a glucose measurement obtained is higher when using whole blood compared to plasma or serum. Use of whole blood allows for more correct assessment and diagnosis, which are necessary for appropriate therapeutic intervention.     

Interobserver reliability of canine urine specific gravity assessed by analog or digital refractometers

Refractometry is utilized routinely to evaluate canine urine specific gravity (USG) in veterinary clinical settings. We aimed to determine if the magnitude of interobserver reliability when assessing canine USG via refractometry could impact clinical judgment. USG was determined in 38 dogs by 3 registered veterinary technicians (RVTs) using both an optical analog refractometer and a digital refractometer. Summary statistics were reported, interobserver reliability was assessed via intraclass correlation coefficient (ICC) analysis through a 2-way mixed-effects model, and agreement between RVT pairs was compared through Bland–Altman plots. The median analog refractometer USG measurement was 1.018 (range: 1.004–1.040) and for the digital refractometer was 1.0176 (1.0035–1.0357). The analog refractometer average measure ICC was 0.995 (95% CI: 0.992, 0.997; p < 0.001). The digital refractometer average measure ICC was 0.999 (95% CI: 0.999, 1.000; p < 0.001). Strong agreement between all pairs of RVTs was seen via Bland–Altman plots for both analog and digital refractometers, with 95% CIs spanning no more than 0.002 in either the positive or negative direction for all pairings. The interobserver variability in canine USG measurements by RVTs was trivial and did not impact clinical judgment and decision-making.     

Central nervous system lesions caused by canine distemper virus in 4 vaccinated dogs

We examined the cerebellum and cerebrum of 4 vaccinated dogs, 3–60-mo-old, that displayed clinical signs of canine distemper virus (CDV) infection, and died 7–40 d after developing neurologic signs. The main histologic lesions were demyelination, gliosis, meningitis, perivascular lymphocytic cuffing, and inclusion bodies. These lesions were similar in all 4 cases regardless of the time since vaccination, except that meningoencephalitis and gliosis were subacute in 3 dogs and chronic in 1 dog. However, these differences did not appear to be related to their vaccination status. Immunohistologically, a CDV-positive immunoreaction was seen mainly in astrocytes, neurons and their axons, lymphocytes around and in the blood vessels of the pia mater and choroid plexus, ependymal cells of each ventricle, and the cells of the choroid plexus. The histologic and immunohistologic changes were similar in the cerebellum and cerebrum. The genetic characterization of the virus strains in 2 of these naturally occurring canine distemper cases confirmed that they were South American wild-type strains (Kiki and Uy251) belonging to the EU1/SA1 lineage. These strains are not included in the commercial CDV vaccines available in Uruguay.     

Effects of storage time on chemistry results from canine whole blood,heparinized whole blood,serum and heparinized plasma

The stability and storage characteristics of 24 blood constituents from dogs including nine enzymes (ALP, ALT, amylase, AST, CK, GGT, GLDH, LDH, lipase), 15 metabolites and minerals (albumin, bile acids, bilirubin, calcium, cholesterol, creatinine, fructosamine, glucose, magnesium, phosphate, potassium, protein, sodium, triglycerides, urea) were studied. Conditions studied included storing of nonanticoagulated and heparinized whole blood for 3 days (Part A), and storing of serum and heparinized plasma for 3 days (Part B). The storage temperature for both studies was +4 degrees C from day 0 to day 1, and +20 degrees C, from day 1 to day 2 and 3. Eight of 24 analytes showed no significant differences (p > 0.05) for three days in whole blood. However, the stability of all 24 analytes greatly improved by storing serum or heparinized plasma compared to nonanticoagulated or heparinized whole blood. In stored serum or heparinized plasma, 20 of 24 analytes showed no significant differences (p < 0.05) for 3 days. Nine of 24 analytes showed significant differences (p < 0.05) between serum and heparinized plasma, where CK, LDH, GGT, and potassium showed differences of possible clinical importance. This study strongly supports the practice of separating serum/plasma from clot/cells as promptly as possible to achieve improved stability for most analytes under test.     

Intra‐operative imaging of surgical margins of canine soft tissue sarcoma using optical coherence tomography

Optical coherence tomography (OCT) is a rapid non‐invasive imaging technique that has shown high sensitivity for intra‐operative surgical margin assessment in human breast cancer clinical trials. This promising technology has not been evaluated in veterinary medicine. The objective of this study was to correlate normal and abnormal histological features with OCT images for surgical margins from excised canine soft tissue sarcoma (STS) and to establish image evaluation criteria for identifying positive surgical margins. Fourteen client‐owned dogs underwent surgical resection of a STS and OCT imaging of 2 to 4 areas of interest on the resected specimen were performed. Following imaging these areas were marked with surgical ink and trimmed for histopathology evaluation. Results showed that different tissue types had distinct characteristic appearances on OCT imaging. Adipose tissue exhibited a relatively low scattering and a honey‐comb texture pattern. Skeletal muscle and sarcoma tissue were both dense and highly scattering. While sarcoma tissue was highly scattering, it did not have organized recognizable structure in contrast to muscle which showed clear fibre alignment patterns. In this investigation, we showed different tissue types had different and characteristic scattering and image texture appearances on OCT, which closely correlate with low‐power histology images. Given the differentiation between tissue types the results support that OCT could be used to identify positive surgical margins immediately following resection of STS. Further research is needed to assess the diagnostic accuracy of this method for surgical margin assessment.     

Prevalence of canine cataract: preliminary results of a cross-sectional study

Objective In this study 2000 dogs were examined ophthalmoscopically to determine presence of cataract. Materials and methods The dogs examined were predominantly from veterinary hospital populations but also from the Waltham Center For Pet Nutrition, rehoming charities and breeding kennels. Prevalence of cataract was thus determined for different age groups (year cohorts). The age at which prevalence of cataract was 50% (C₅₀) was determined indirectly from a fitted prevalence curve. Results The mean ± standard deviation of C₅₀ for all dogs in the study was 9.4 ± 3.3 years. All dogs over 13.5 years were affected by some degree of lens opacity. C₅₀ was determined for animals of different genders and different breeds. For dogs of six breeds sufficient data were available for calculation of breed‐specific C₅₀. In these dogs C₅₀ was positively correlated with longevity with a least squares correlation coefficient of 0.74. Conclusion The study yields novel findings regarding the prevalence and incidence of cataract in the dog and forms the basis for considerable further work on the epidemiology and pathophysiology of age‐related cataract in the dog.     

Tibial tuberosity advancement in 92 canine stifles: initial results,clinical outcome and owner evaluation

Objective To investigate the clinical outcomes, complications and owners' evaluation of the tibial tuberosity advancement (TTA) procedure in canine stifles. Methods A retrospective study of hospital records was performed to identify dogs diagnosed with partial or complete cranial cruciate ligament (CCL) rupture that had undergone TTA repair. Information obtained included signalment, period of lameness, surgical report, evidence of meniscal injury, postoperative recovery and peri-operative complications. Owners were asked to assess the long-term outcome. Results In a total of 72 dogs (median age, 6 years; median body weight, 34.8 kg), TTA was performed in 92 stifles. Twenty breeds were represented, with Labrador Retrievers and Rottweilers the most common. The period of lameness ranged from 3 days to 24 months. The median pre-operative lameness score was 3/4 and meniscal injury was present in 51 stifles. Minor complications occurred in 29% of cases. Major complications occurred in 6.5% of cases and consisted of meniscal injury and two tibial tuberosity fractures. All were successfully managed, with good limb function when subsequently assessed. In the owner evaluation, 96% reported moderate to great improvement postoperatively, with no lameness at rest and mild to no lameness after vigorous exercise. Conclusion Clinical outcome and owner evaluations in this case series indicate favourable results can be expected when CCL-deficient stifles are treated with TTA.     

Identification of neoplastic cells in blood using the Sysmex XT‐2000iV: a preliminary step in the diagnosis of canine leukemia

Background: Classification of leukemias requires specialized diagnostic techniques. Automated preliminary indicators of neoplastic cells in blood would expedite selection of appropriate tests. Objective: The objective of this study was to assess the capacity of the Sysmex XT‐2000iV hematology analyzer to identify neoplastic cells in canine blood samples. Methods: Blood samples (n=160) were grouped into 5 categories: acute leukemia (n=30), chronic leukemia (n=15), neoplasia without blood involvement (n=41), non‐neoplastic reactive conditions (n=31), and healthy dogs (n=43). WBC counts, WBC flags, scattergrams, percentages of cells with high fluorescence intensity, and percentages of cells in the lysis‐resistant region were evaluated alone or in combination to establish a “leukemic flag.” Sensitivity, specificity, negative (LR?) and positive (LR+) likelihood ratios, and the number of false‐negative (FN) and false‐positive (FP) results were calculated, and receiver operating characteristic curves were designed for numerical values. Results: Among single measurements and parameters, only the evaluation of scattergrams minimized FN and FP results (sensitivity 100%, specificity 94.8%, LR+ 19.17, and LR? 0.00), although their interpretation was subjective. The more objective approach based on the generation of a “leukemic flag” had a sensitivity of 100%, specificity of 87.0%, LR? of 0.00, and LR+ of 7.67. Conclusion: Using a novel gating strategy the Sysmex XT‐2000iV may be used effectively to screen canine blood for hematopoietic neoplasia.     

Validation study of canine urine cortisol measurement with the Immulite 2000 Xpi cortisol immunoassay

We report here validation of the Immulite 2000 Xpi cortisol immunoassay (Siemens; with kit lot numbers <550) for measurement of urine cortisol in dogs, with characterization of the precision (CV), accuracy (spiking-recovery [SR] bias), and observed total error (TEo = bias + 2CV) across the reportable range. Linearity assessed by simple linear regression was excellent. Imprecision, SR bias, and TEo increased markedly with decreasing urine cortisol concentration. Interlaboratory comparison studies determined range-based (RB) bias and average bias (AB). The 3 biases (SR, RB, and AB) and resulting TEo differed markedly. At 38.6 and 552 nmol/L (1.4 and 20 μg/dL), between-run CVs were 10% and 4.5%, respectively, and TEo_RB were ~30% and 20%, respectively, similar to observations in serum in another validation study. These analytical performance parameters should be considered for urine cortisol:creatinine ratio (UCCR) result interpretation, given that, for any hypothetical errorless urine creatinine measurement, the error % on UCCR mirrors the error % on urine cortisol. Importantly, there is no commonly used interpretation threshold for UCCR, given that UCCR varies greatly depending on measurement methods and threshold computation. To date, there is no manufacturer-provided quality control material (QCM) with target values for urine cortisol with an Immulite; for Liquicheck QCM (Bio-Rad), between-run imprecision was ~5% for both QCM levels. Acceptable QC rules are heavily dependent on the desired total allowable error (TEa) for the QCM system, itself limited by the desired clinical TEa.     

Single‐voxel and multi‐voxel spectroscopy yield comparable results in the normal juvenile canine brain when using 3 Tesla magnetic resonance imaging

Conventional magnetic resonance imaging (MRI) characteristics of canine brain diseases are often nonspecific. Single‐ and multi‐voxel spectroscopy techniques allow quantification of chemical biomarkers for tissues of interest and may help to improve diagnostic specificity. However, published information is currently lacking for the in vivo performance of these two techniques in dogs. The aim of this prospective, methods comparison study was to compare the performance of single‐ and multi‐voxel spectroscopy in the brains of eight healthy, juvenile dogs using 3 Tesla MRI. Ipsilateral regions of single‐ and multi‐voxel spectroscopy were performed in symmetric regions of interest of each brain in the parietal (n = 3), thalamic (n = 2), and piriform lobes (n = 3). In vivo single‐voxel spectroscopy and multi‐voxel spectroscopy metabolite ratios from the same size and multi‐voxel spectroscopy ratios from different sized regions of interest were compared. No significant difference was seen between single‐voxel spectroscopy and multi‐voxel spectroscopy metabolite ratios for any lobe when regions of interest were similar in size and shape. Significant lobar single‐voxel spectroscopy and multi‐voxel spectroscopy differences were seen between the parietal lobe and thalamus (P = 0.047) for the choline to N‐acetyl aspartase ratios when large multi‐voxel spectroscopy regions of interest were compared to very small multi‐voxel spectroscopy regions of interest within the same lobe; and for the N‐acetyl aspartase to creatine ratios in all lobes when single‐voxel spectroscopy was compared to combined (pooled) multi‐voxel spectroscopy datasets. Findings from this preliminary study indicated that single‐ and multi‐voxel spectroscopy techniques using 3T MRI yield comparable results for similar sized regions of interest in the normal canine brain. Findings also supported using the contralateral side as an internal control for dogs with brain lesions.     

Cardiac‐gated,phase contrast magnetic resonance angiography is a reliable and reproducible technique for quantifying blood flow in canine major cranial abdominal vessels

Blood flow changes in cranial abdominal vessels are important contributing factors for canine hepatic disease. This prospective, experimental, pilot study aimed to evaluate cardiac‐gated, phase contrast magnetic resonance angiography (PCMRA) as a method for characterizing blood flow in canine major cranial abdominal vessels. Eleven, healthy, adult beagle dogs were sampled. Cardiac‐gated, phase contrast magnetic resonance angiography of the cranial abdomen was performed in each dog and blood flow was independently measured in each of the major cranial abdominal vessels by three observers, with two observers recording blood flow values once and one observer recording blood flow values three times. Each dog then underwent ultrasonographic examination of the liver with fine needle aspirations and biopsies submitted to cytologic and histologic examination. The mean absolute stroke volume and velocity were respectively 9.6 ± 1.9 ml and –11.1 ± 1.1 cm/s for the cranial abdominal aorta, 2.1 ± 0.6 ml and –6.6 ± 1.9 cm/s for the celiac artery, and 2.3 ± 1.0 ml and –7.9 ± 3.1 cm/s for the cranial mesenteric artery. The mean absolute stroke volume and velocity were respectively 6.7 ± 1.3 ml and 3.9 ± 0.9 cm/s for the caudal vena cava and 2.6 ± 0.9 ml and 3.2 ± 1.2 cm/s for the portal vein. Intraobserver reliability was excellent (intraclass correlation coefficient > 0.9). Interobserver reproducibility was also excellent (intraclass correlation coefficient 0.89–0.99). Results of liver ultrasonography, cytology, and histopathology were unremarkable. Findings indicated that cardiac‐gated, phase contrast magnetic resonance angiography is a feasible technique for quantifying blood blow in canine major cranial abdominal vessels. Blood flow values from this sample of healthy beagles can be used as background for future studies on canine hepatic disease.