Leukocyte Esterase and Nitrite Urine Reagent Strip Utility Under Altered Assay Conditions in Dogs.

The study evaluated leukocyte esterase (LE) and nitrite reagent pad usefulness in canine urine using multiple time endpoints and decision thresholds. Whole and sedimented urine samples from 116 client-owned dogs were analyzed with reagent strips from four manufacturers. Results for LE and nitrite were recorded every 30 s up to 150 s using "trace" and "+" positive thresholds and compared to microscopic urine sediment analysis and culture results. Sedimented urine assessed at 150 s with a "trace" positive threshold for LE had significantly or trending higher sensitivity (brand dependent) for pyuria detection compared with manufacturer recommendations. Using these parameters, LE sensitivity and negative likelihood ratio were 66.67-89.47%, and 0.13-0.37, respectively. Following manufacturer recommendations, LE specificity and positive likelihood ratio were 96.91-100% and 17.02-30.63, respectively. Nitrite performance for pyuria and bacteriuria detection was poor for all conditions. LE reagent pads are a useful screening test for pyuria detection using sedimented urine and a "trace" positive threshold at 150 s, whereas LE use with traditional manufacturer recommendations is a good confirmatory test for pyuria detection. Nitrite reagent pads are not useful for pyuria or bacteriuria detection.

Synovial Cysts and Myxomas in 16 Cats.

This report describes the clinical and pathologic characteristics of cystic and myxomatous lesions of synovial joints in 16 cats. The average age was 13.4 years. The elbow was most commonly affected (12/16), and all lesions were unilateral. Degenerative joint disease was a frequent concurrent but bilateral condition. The lesions consisted of fluid-filled cysts lined by synoviocytes (3 cases), solid foci of stellate cells in a myxomatous matrix (2 cases), or a combination of the two (11 cases). In some cases there were areas of transition between the cystic and myxomatous lesions. Mitoses and other features of malignancy were rare to nonexistent. In the 13 cats with follow-up information, the lesion gradually increased in size over a period of years. None of the cats died or were euthanized because of this lesion. We propose that some cats with degenerative joint disease develop synovial cysts, which have the potential to transform to a synovial myxoma.

Laboratory diagnostics, phylogenetic analysis and clinical outcome of a subcutaneous Mycoleptodiscus indicus infection in an immunocompetent cat.

BACKGROUND: Mycoleptodiscus indicus is a dematiaceous hyphomycete fungus found on plant leaves. It has been rarely reported as a cause of human or animal disease, possibly because it is difficult to culture and identify from clinical specimens. Infections are presumably acquired by traumatic implantation. CASE PRESENTATION: An 8-year-old non-immunosuppressed cat from Georgia, USA, presented with a left front leg swelling without lameness. Cytology from a fine needle aspirate revealed pyogranulomatous inflammation with both cytoplasmic and extracellular fungal elements. There were septate hyphae with irregularly sized segments, non-staining uneven walls, and rounded yeast-like forms from which longer hyphae arose in a hub-and-spoke pattern. A mold was isolated on agar from a fine needle aspirate collected 1 week later and identified as M. indicus by morphology, DNA sequencing and phylogenetic analysis. The cat recovered completely and uneventfully with antifungal treatment. CONCLUSIONS: We report a previously undescribed presentation of M. indicus causing a subcutaneous infection in a cat with successful antifungal treatment. In this study we highlight the potential of M. indicus to infect immunocompetent animals, and the veterinary medical community should be aware of its unusual but characteristic clinical, microbiological and cytologic presentation.

Impact of photographer experience and number of images on telecytology accuracy.

BACKGROUND: Studies evaluating the potential impact of photographer experience or the number of images evaluated using the "store-and-forward" method of telecytology are not reported. OBJECTIVES: This study aimed to determine the diagnostic sensitivity (Se) and specificity (Sp) of static telecytology when images were taken by experienced and inexperienced cytologists and when the number of images taken varied. Clinical agreement between the diagnoses was compared. METHODS: Fifty archived cytology cases were randomly chosen. A board-certified clinical pathologist and a recent veterinary graduate took five images of each case. A third pathologist made a preliminary diagnosis after reviewing two images, and a final diagnosis after reviewing all images. The gold standard for comparison was the glass slide cytologic diagnosis. RESULTS: Se and Sp were higher for the experienced cytologist and the evaluation of more images, but differences were not statistically significant. Clinical agreement between the image and glass slide diagnoses was significantly higher when images were taken by an experienced rather than inexperienced cytologist after the evaluation of two (P = .007) and five images (P = .008). The telecytology diagnoses agreed with the gold standard diagnoses more frequently after evaluation of five images rather than two when images were captured by both the experienced (P < .001) and inexperienced cytologist (P < .001). CONCLUSIONS: There is more clinical agreement when the photographer has more cytology experience and when more images are provided for interpretation.

Clinicopathologic Features, Diagnosis, and Characterization of the Immune Cell Population in Canine Choroid Plexus Tumors.

The World Health Organization characterizes human choroid plexus tumor (CPT) as papilloma (CPP), atypical CPP (ACPP), and carcinoma (CPC). CPCs can disseminate via cerebrospinal fluid and be mistaken for metastatic carcinoma, creating a diagnostic challenge. Kir7.1 immunohistochemistry (IHC) is a highly reliable tool for diagnostic confirmation of CPTs and their differentiation from metastatic carcinomas in human beings and dogs. This study describes the neuropathology, Kir7.1 staining profile, and the immune cell population within the tumor microenvironment in 11 CPTs in dogs. Archived tissue sections with a diagnosis of CPT were examined and immunolabelled with Kir7.1 for diagnostic confirmation. The number of Ki67-positive neoplastic cells was calculated in 2.4 mm2 (equivalent to 10 FN22/40X fields), and a mean value was generated for each neoplasm. IHC for CD3, CD20, MAC387, and Iba1 was performed for immune cell characterization, and the number of stained cells for each antibody was counted in 2.4 mm2, generating individual cumulative values for each antibody. T-tests with Bonferroni correction evaluated IHC differences between tumor types, and Spearman's rank correlations evaluated relationships among IHC markers. Kir7.1 immunoreactivity was intense at the apical cell membrane in CPPs and ACPPs, and at the apical cell membrane and cytoplasm in CPCs. Ki67 immunoreactivity was detected in all cases. CD3+ and CD20+ lymphocytes trended together (p = 0.005) and were present within and around all CPTs. Five cases had intravascular MAC387+ monocytes. Iba1 immunoreactivity was robust within and around all tumors. Statistical differences in immune cell markers were not found among tumor types. As previously reported, Kir7.1 is a reliable antibody for the diagnosis of canine CPTs. Although immune cells were present in all cases, no significant associations were found between the type of cells and tumor diagnosis. The characterization of the immune cells within CPTs could be useful in future studies involving immunotherapy.

Evaluation of a Home Urinalysis Kit in Dogs.

Dry reagent strip evaluation of urine is a standard screening and diagnostic test used to assess overall health and help detect or rule out specific disease conditions. A commercial at-home urinalysis reagent strip kit using a smartphone app to evaluate free-catch urine is being marketed directly to dog and cat owners. We compared agreement between simultaneous urinalysis using the commercial kit and standard reference methods in 48 canine urines submitted to our referral laboratory. Agreement was defined by analyte based on clinical impact. Sensitivity, specificity, and Cohen's kappa evaluated categorical data, and a paired t test was used for continuous variables (significance P < .05). The commercial kit had ≥1 disagreement with the reference method per sample and produced results if the test strip was absent or reversed. Specific gravity and pH concurred with the reference method in only 31% (P < .011) and 27% (P < .001) of cases, respectively. The sensitivity was low for all analytes except ketones, which had 77% false positives. False-positive nitrites and leukocytes were also frequent (36 and 19%, respectively). False negatives for blood (27%), nitrites (38%), and protein (54%) were common. This kit is inaccurate; its use for clinical decisions is not recommended.

Outcomes of and risk factors for presumed canine H3N2 influenza virus infection in a metropolitan outbreak.

OBJECTIVE To determine clinical signs, case fatality rate, and factors associated with positive results of PCR testing for canine influenza virus (CIV) in dogs during an H3N2 CIV outbreak in the Atlanta area. DESIGN Cross-sectional study. ANIMALS 220 dogs with a nasal swab specimen submitted to an Atlanta-area diagnostic laboratory between May 1 and July 2, 2015, for PCR assay detection of CIV specifically or CIV and 5 other respiratory pathogens. PROCEDURES Veterinarians of tested dogs were surveyed by various means to collect information regarding clinical signs, survival status at the time of survey completion, vaccination history (≤ 12 months prior to testing), and travel history (≤ 2 months prior to testing). Data were compared between CIV-positive and CIV-negative dogs. RESULTS Surveys for 120 (55%) dogs were completed. Forty (33%) of these dogs had positive results of CIV testing. No significant differences were identified between CIV-positive and CIV-negative dogs regarding breed, sex, reproductive status, duration of clinical signs prior to testing, other dogs in the household, or travel history. When other factors were controlled for, CIV-positive dogs were more likely to be adult (> 1 year of age) than juvenile (≤ 1 year of age) and to be inappetent. Only 1 (3%) CIV-positive dog died during the study period (shortly after it was evaluated because of respiratory signs). CONCLUSIONS AND CLINICAL RELEVANCE From May 1 to July 2, 2015, the reported clinical signs of dogs tested during the Georgia H3N2 CIV outbreak were similar to those reported for dogs with H3N8 CIV infection, and the case fatality rate was low.

Demonstration of the ability of a canine Lyme vaccine to reduce the incidence of histological synovial lesions following experimentally-induced canine Lyme borreliosis.

Lyme disease in dogs can be effectively prevented by vaccination against antigens expressed by the spirochete Borrelia burgdorferi during transmission by the tick vector Ixodes sp. Lyme vaccine efficacy has traditionally been based on indicators of infection following wild-caught tick challenge whereas most other types of vaccine are required to demonstrate protection from clinical signs of disease. In this vaccination-challenge study we sought to demonstrate the ability of a nonadjuvanted, outer surface protein A (OspA) vaccine to protect from infection and to prevent synovial lesions consistent with Borreliosis. Thirty, purpose-bred beagles were randomly divided into vaccinated and unvaccinated groups. The vaccinated group was administered two subcutaneous doses of a nonadjuvanted, purified, Borrelia burgdorferi OspA vaccine at a 21- day interval. Dogs were challenged by wild-caught, B. burgdorferi-infected ticks (Ixodes scapularis). Clinical signs, serology, Borrelia isolation and PCR evaluated antemortem vaccine efficacy. Postmortem histopathological analysis of synovial tissue was compared to antemortem infection status. Borreliosis was demonstrated by Borrelia isolation from skin biopsies in 13 out of 15 unvaccinated dogs. All unvaccinated dogs' Western blot profiles were consistent with infection. Two of 15 vaccinated dogs had at least one positive spirochete culture which cleared 91days post-challenge, and Western blot profiles were consistent with vaccination alone. No dogs, vaccinated or unvaccinated, exhibited clinical signs consistent with borreliosis. Based on a histopathological cumulative joint scoring system (CJS), all unvaccinated dogs had synovial lesions indicative of Lyme disease. Only one of the vaccinated dogs had a CJS that was greater than the statistical cut off score for the absence of synovial lesions. There was high correlation between clinical histopathology and spirochete isolation. Infection with B burgdorferi may produce inconsistent clinical signs of lameness. Histopathological changes in joints from infected dogs are reliable indicators of borreliosis and correlate well with other indicators of infection. This model provides support that vaccination with a nonadjuvanted, purified OspA vaccine offers protection from Borrelia infection and the resulting synovial lesions that can lead to clinical signs of lameness.

Evaluation of cytospin precision in low cellularity canine cerebrospinal fluid.

The cell count and differential of cerebrospinal fluid (CSF) cytologic examination classify CSF as inflammatory or not. The cytospin cell yield is related to cell count, but to our knowledge a relationship has not been characterized and cytospin precision is undocumented in any species. The objective of our study was to calculate intra-assay precision of cellular yield and differential on cytocentrifuged canine CSF, determine the factors that may affect precision, and predict the number of cytospins necessary to confirm mild neutrophilic pleocytosis. Ten concurrent replicate cytospins were created from nonhemorrhagic CSF, obtained from 60 dogs in other terminal studies, with either a manual or calibrated pipetting technique. Up to 500 cells per cytospin were counted and classified on each slide. Coefficient of variation (CV), multiple regression, and probabilities were calculated for relationships between cell yield and independent factors including technique, total nucleated cell count, cell differential, and total protein. Manual and calibrated pipetting had similar CVs (average 31%) for total cell yield, but the calibrated technique had fewer foamy macrophages. CV for neutrophil percentage among low cellularity samples with any neutrophils was 146%. Probability based on linear regression showed that 1 cytospin is sufficient to identify samples with >3% neutrophils. Occasional neutrophils, eosinophils, mitotic figures, phagocytic cells, and ependymal cells were seen in many low cellularity canine CSF samples. Canine CSF cytospin cell yield and differential evaluations are imprecise. Calibrated rather than manual pipetting is recommended.

Factors influencing survivorship of rehabilitating green sea turtles (Chelonia mydas) with fibropapillomatosis.

Marine turtle fibropapillomatosis (FP) is a debilitating, infectious neoplastic disease that has reached epizootic proportions in several tropical and subtropical populations of green turtles (Chelonia mydas). FP represents an important health concern in sea turtle rehabilitation facilities. The objectives of this study were to describe the observed epidemiology, biology, and survival rates of turtles affected by FP (FP+ turtles) in a rehabilitation environment; to evaluate clinical parameters as predictors of survival in affected rehabilitating turtles; and to provide information about case progression scenarios and potential outcomes for FP+ sea turtle patients. A retrospective case series analysis was performed using the medical records of the Georgia Sea Turtle Center (GSTC), Jekyll Island, Georgia, USA, during 2009-2013. Information evaluated included signalment, morphometrics, presenting complaint, time to FP onset, tumor score (0-3), co-morbid conditions, diagnostic test results, therapeutic interventions, and case outcomes. Overall, FP was present in 27/362 (7.5%) of all sea turtles admitted to the GSTC for rehabilitation, either upon admittance or during their rehabilitation. Of these, 25 were green and 2 were Kemp's ridley turtles. Of 10 turtles that had only plaque-like FP lesions, 60% had natural tumor regression, all were released, and they were significantly more likely to survive than those with classic FP (P = 0.02 [0.27-0.75, 95% CI]). Turtles without ocular FP were eight times more likely to survive than those with ocular FP (odds ratio = 8.75, P = 0.032 [1.21-63.43, 95% CI]). Laser-mediated tumor removal surgery is the treatment of choice for FP+ patients at the GSTC; number of surgeries was not significantly related to case outcome.

Spontaneous chronic T-cell leukemia in a male rhesus macaque ( Macaca mulatta).

Blood smears from a 24-year-old male rhesus macaque ( Macaca mulatta) used for cognitive function studies were evaluated. The macaque had an 8-month history of gradual weight loss and increasing lymphocytosis. Most of the lymphocytes present were small to medium and had a mature morphology. Based on the degree and duration of the lymphocytosis, and the appearance of the lymphocytes, a diagnosis of chronic lymphocytic leukemia was made. The animal tested negative for 4 viral diseases that are commonly associated with lymphoproliferative disorders in Old World monkeys. Over the course of 12 months, the lymphocytosis progressed from 18.4 to 384 × 103 lymphocytes/µl (reference range: 0.8-17 × 103 cells/µl), and euthanasia was elected. On histologic examination, cluster of differentiation (CD)3- and CD8-positive, CD79-negative neoplastic cells comprised 40-60% of the bone marrow, diffusely obscured the normal splenic architecture, and were present in the vascular channels in other organs. Findings were characteristic of T-cell lymphocytic leukemia. Naturally occurring T-cell lymphocytic leukemia has been rarely reported in rhesus macaques and, to the authors' knowledge, never in males. A persistent lymphocytosis characterized by a monomorphic population of CD3- and CD8-positive cytotoxic T-lymphocytes and the presence of neoplastic cells in the bone marrow led to a diagnosis in the current case.

Sonoclot evaluation of whole blood coagulation in healthy adult dogs.

OBJECTIVE: To establish a standard protocol for analysis of canine whole blood and generate reference intervals for healthy dogs using the Sonoclot analyzer, and to compare Sonoclot values to standard and viscoelastic coagulation tests. DESIGN: Prospective study. SETTING: Veterinary University research facility and teaching hospital. ANIMALS: Twelve healthy random source dogs and 52 healthy dogs from the general veterinary school population. INTERVENTIONS: Blood sampling for viscoelastic coagulation testing. MEASUREMENTS AND MAIN RESULTS: Blood was collected from 12 healthy adult dogs by jugular venipuncture. After a rest period at room temperature of 30, 60, or 120 minutes, 340 µL of citrated blood was added to 20 µL of 0.2 M CaCl(2) in 1 of 2 cuvette types warmed to 37° C. Cuvettes contained a magnetic stir-bar with glass beads (gbACT+) or only a magnetic stir-bar (nonACT). Reference interval samples were collected from 52 healthy adult dogs and analyzed in duplicate. The ACT, CR, and PF were not affected by duration of rest period for either cuvette type. ACT variability was decreased when using gbACT+ cuvettes (P < 0.05). In normal dogs reference intervals (mean ± 2 SD) using gbACT+ cuvettes were: ACT 56.0-154.0 seconds, CR 14.85-46.0, and PF 2.1-4.05. ACT correlated to TEG R-time, K-time, and angle, while CR correlated with all TEG parameters. Fibrinogen correlated with ACT, CR, and PF. Sonoclot did not correlate with other common coagulation tests. CONCLUSIONS: Sonoclot provides viscoelastic evaluation of canine whole blood coagulation and correlated to several TEG parameters and fibrinogen. A standard protocol and reference intervals were established.

Identification of altered MicroRNA expression in canine lymphoid cell lines and cases of B- and T-Cell lymphomas.

Canine lymphoma is a common spontaneous tumor with many similarities to human lymphoma, and thus has potential to be an important animal model of lymphomagenesis. This study determined that microRNA (miRNA) expression in canine tumors can be assessed using a commercially available human cancer miRNA qPCR array. miRNA expression in six different canine lymphoid cell lines and in naturally occurring canine B- and T-cell lymphomas was compared using RNA harvested from normal canine peripheral blood mononuclear cells (PBMC) and normal lymph nodes (LN) as controls. We found that false discovery rate (FDR) correction for multiple testing after quantile normalization controlled for variation across arrays and that they were the best methods for normalization and statistical analysis. Increases in miRNAs known to upregulate oncogenes (miR19a+b, miR17-5p) and decreased expression of miRNAs with tumor suppressor functions (miR-203, miR-218, and miR-181a) also seen in human lymphoid malignancies were observed. However, there were few similarities between canine groups. The results of this study indicate that the use of both PBMC and LN cells as controls provides different, but potentially equally important targets for further analysis. Our findings of miRNA dysregulation in canine lymphoid cell lines and clinical cases of lymphoma emphasize the potential of canine lymphoma as an important spontaneous, large animal model of human B- and T-cell lymphomas.

Molecular and pathological investigations of the central nervous system in Borrelia burgdorferi-infected dogs.

Although neurological signs have been reported sporadically in dogs with systemic Lyme disease, it is unknown if neuroborreliosis occurs in dogs. The current study systematically evaluates canine brains for evidence of Borrelia burgdorferi infection. Twelve Beagles were experimentally challenged with B. burgdorferi-infected ticks at 18 weeks of age, and 2 unexposed dogs served as controls. One of the uninfected dogs and 6 infected dogs were each given 5 daily immunosuppressive doses of dexamethasone starting at 153 days post-infection. Eleven dogs were confirmed as infected by skin punch biopsy polymerase chain reaction (PCR) and serology. Neurological signs were not seen in any dogs through the end of the 190-day study. Whole blood, serum, cerebrospinal fluid (CSF), and brains from all dogs were collected. DNA was extracted from blood, CSF, and brain and evaluated by PCR for B. burgdorferi. Formalin-fixed brain tissue was examined by histopathology, immunohistochemistry, and PCR. Immunohistochemical staining for B. burgdorferi antigen was negative in all cases. The CSF analysis was normal, and PCR was uniformly negative for B. burgdorferi in all dogs. Six of the 11 (45%) infected dogs had mild to moderate lymphoplasmacytic choroid plexitis, which was more pronounced in the immunosuppressed dogs. The lack of B. burgdorferi DNA and immunohistochemical evidence of organisms, including within the choroid plexus lesions, suggests that B. burgdorferi does not have a direct role in the etiopathogenesis of canine central nervous system pathology.

Panniculitis attributable to Mycobacterium goodii in an immunocompetent dog in Georgia.

CASE DESCRIPTION: A 5-year-old 38.3-kg (84.5-lb) mixed-breed dog was examined because of acute onset of lethargy and anorexia. Four days later, a raised, firm, warm 15 × 10-cm lesion was detected in the right caudal paralumbar area. CLINICAL FINDINGS: Cephalexin treatment yielded a poor response. Formalin-fixed tissue and fluid samples from the cystic areas of the lesion were submitted for cytologic and histologic examinations, routine bacterial and mycobacterial culture, and genus identification and 16S partial sequencing via PCR assays. Cytologic examination revealed chronic pyogranulomatous inflammation. Histologic examination by use of routine, Giemsa, silver, acid-fast, and modified acid-fast stains revealed multifocal nodular granulomatous panniculitis without identifiable organisms. Mycobacteria were initially identified via PCR assay and mycobacterial culture within 3 days. Mycobaterium goodii was speciated by use of partial 16S RNA sequence analysis. TREATMENT AND OUTCOME: The lesion resolved after long-term treatment with a combination of rifampin and clarithromycin and insertion of a Penrose drain. There has been no recurrence of the condition. CLINICAL RELEVANCE: M goodii is an environmental rapidly growing mycobacterium and is a zoonotic pathogen. Infections have not been previously reported in domestic animals in North America, although there are rare reports of infection in humans associated with surgery, especially surgical implants. Domestic animals are a potential sentinel for this non-tuberculous mycobacterial infection in humans, although lack of speciation in infections of domestic animals likely underestimates the potential public health importance of this pathogenic organism. Current microbiological molecular methods allow for a rapid and inexpensive diagnosis.

Hematology and plasma chemistry reference intervals for mature laboratory pine voles (Microtus pinetorum) as determined by using the nonparametric rank percentile method.

Plasma biochemical and hematologic values are important parameters for assessing animal health and experimental results. Although normal reference values for many rodent species have been published, there is a dearth of similar information for the genus Microtus. In addition, most studies use a mean and standard deviation to establish reference intervals, but doing so is not the recommendation of the Clinical and Laboratory Standards Institute (formerly the National Committee on Clinical Laboratory Standards) or the International Federation of Clinical Chemistry and Laboratory Medicine. The purpose of this study was to establish normal reference parameters for plasma biochemistry and hematology in mature pine voles (Microtus pinetorum) by using the nonparametric rank percentile method as recommended by the 2 laboratory medicine organizations mentioned. Samples of cardiac blood from a closed colony of pine voles were collected at euthanasia and evaluated under rodent settings on 2 automated hematology analyzers from 2 different manufacturers and on the same type of automated biochemistry analyzer. There were no sex-associated clinically significant differences between the sexes; younger animals had a lower hematocrit, higher mean corpuscular volume, and lower mean corpuscular hemoglobin concentration than did older animals. Only platelet counts differed when comparing hematologic values from different analyzers. Relative to rats and mice, pine voles have a lower mean corpuscular volume and higher red blood cell count, higher blood urea nitrogen, much higher alanine aminotransferase, and lower glucose and phosphorous concentrations. Hematology and plasma biochemical results obtained in this study are considered representative for healthy adult laboratory pine voles under similar environmental conditions.