Research Note: Comparison of chicken blood chemistry and electrolyte parameters between the portable i-STAT1 clinical analyzer and VetScan VS2 serum biochemistry panel using Hy-Line commercial white-egg laying hens.

The i-STAT1 clinical analyzer has become an increasingly popular tool in clinical production animal medicine as it can provide pen-side results in a cost effective and timely manner when compared to standard benchtop serum biochemistry blood gas and chemistry analyses. This study compares the results of the portable Abbott i-STAT1 analyzer and the Abaxis VetScan VS2 for glucose (Glu, mg/dL), ionized Ca (mmol/L), Na (mmol/L), and K (mmol/L) values. Three genetically distinct commercial varieties (CV) of Hy-Line white-egg laying hens are used in this study (Hy-Line W-36, Hy-Line W-80, and Hy-Line W-80+). Thirty blood samples (n = 10 per CV) were obtained in the production house from the brachial vein and concurrently analyzed by the i-STAT1 portable device. Serum from 22 of these same samples was analyzed via VetScan VS2, a benchtop serum clinical biochemistry analyzer, using VetScan Avian/Reptilian Profile Plus reagent rotors. A paired T-test was used to test for statistical differences in means between the 2 instruments for each of the parameters. Parameters with significant mean differences were then subject to correlation and regression analysis to further evaluate relationships between the results from the 2 methods. Significant differences between means were found for Glu, Na, and K levels. Ca levels were found to be not directly comparable by the 2 analysis instruments. This comparison elucidates the importance of clinical analyzer validations when applying different strategies of diagnostic medicine in poultry.

Clinical presentation, cardiovascular findings, etiology, and outcome of myocarditis in dogs: 64 cases with presumptive antemortem diagnosis (26 confirmed postmortem) and 137 cases with postmortem diagnosis only (2004-2017).

INTRODUCTION: This study describes presentation, cardiovascular abnormalities, etiology, and outcome of canine myocarditis in geographic areas not endemic for Trypanosoma or Leishmania. ANIMALS: Sixty-four (presumed antemortem diagnosis) and 137 (postmortem diagnosis only) client-owned dogs at two tertiary care facilities were included. MATERIALS AND METHODS: Medical records of dogs with clinical or histopathological diagnosis of myocarditis were reviewed retrospectively. RESULTS: Common examination findings in dogs with a presumed antemortem diagnosis included fever (21%) and heart murmur (19%). Median cardiac troponin I was 12.2 ng/mL (range: 0.2-808.0 ng/mL), and troponin exceeded 1.0 ng/mL in 26 of 29 (90%) dogs. Ventricular ectopy was the most common arrhythmia (54%), whereas decreased left ventricular systolic function was the most common echocardiographic abnormality (56%). An infectious etiology was diagnosed in 35 of 64 (55%) dogs. Confirmed infectious etiologies included bacterial sepsis (n = 9) or extension of endocarditis (3), toxoplasmosis or neosporosis (3), parvovirus (2), and one case each of bartonellosis, trypanosomiasis, leptospirosis, and dirofilariasis. Median survival time was 4 days (range: 0-828 days) for all dogs vs. 82 days for dogs who survived at least 2 weeks after diagnosis. Presence of pericardial effusion or azotemia was a significant predictor of non-survival. The most common inflammatory infiltrate on histopathology was neutrophilic (47%), and 20 of 137 (14.5%) dogs had concurrent bacterial endocarditis on postmortem. CONCLUSIONS: Bacterial infection was the most common confirmed etiology of myocarditis in this study. Prognosis for canine myocarditis is guarded and similar to that reported for infective endocarditis. Criteria for the antemortem diagnosis of canine myocarditis are suggested.

The association between sympatho-vagal balance and central blood pressures.

PURPOSE: High central blood pressure is more predictive of cardiovascular disease (CVD) versus high peripheral blood pressure. Measures of central pressures (CPs) include, central systolic blood pressure (CSBP) and central diastolic blood pressure. Measures of central pressures augmentation (CPsA) include augmentation pressure (AP) and the augmentation index @ 75 beats·min-1 (AIx@75). Increased sympathetic tone (ST) is also associated with CVD. The low to high frequency ratio (LF/HF) is often used to determine sympatho-vagal balance. Given the association between ST, CPs, CPsA and CVD there is a need to understand the association between these predictors of CVD. The aims of this study were to examine the association between the LF/HF ratio, CPs, and CPsA in men and women collectively and based on gender. METHODS: We measured the LF/HF ratio, CSBP, AP, and AIx@75 in 102 participants (41F/61M). The LF/HF ratio was determined via power spectral density analysis. CSBP, AP, and AIx@75 were determined via applanation tonometry. RESULTS: The LF/HF ratio was inversely associated with AP (r = -0.26) and AIx @75 (r = -0.29) in the combined group of men and women. The LF/HF ratio was inversely associated with CSBP (r = -0.27), AP (r = -0.28), and AIx@75 (r = -0.32) in men, but not in women. CONCLUSION: There is an inverse association between the LF/HF ratio, AP, and AIx@75 in men and women combined. The association between the LF/HF ratio, CSBP, AP, and AIx@75, differs based on gender.

Establishment of Hy-Line commercial laying hen whole blood gas and biochemistry reference intervals utilizing portable i-STAT1 clinical analyzer.

Blood gas and biochemistry reference intervals were established for 3 genetically distinct commercial varieties (CVs) of Hy-Line laying hens: 2 brown-egg layers (Hy-Line Brown, Hy-Line Silver Brown) and a tint-egg layer (Hy-Line Sonia) utilizing the i-STAT1 analyzer. Each respective variety of laying hen was sampled on a replicate cycle of 2 wk for a total of 6 replicates (35 to 46 wk of age). Blood samples were obtained in the production house from the brachial vein, and subsequently analyzed by the i-STAT1 portable device. i-STAT1 clinical analyzer reports blood gas and biochemistry values for the following parameters: pH, partial pressure of carbon dioxide (pvCO2, mm Hg), partial pressure of oxygen (pvO2, mm Hg), bicarbonate (HCO3, mmol/L), base excess (BE, mmol/L), saturation of oxygen on hemoglobin (sO2%), glucose (Glu, mg/dL), sodium (Na, mmol/L), potassium (K, mmol/L), total concentration of carbon dioxide (TCO2, mmol/L), ionized calcium (iCa, mmol/L), hematocrit (Hct % packed cell volume [PCV]), hemoglobin (Hb, g/dL). A total of 1,800 individual hen i-STAT1 records were utilized in the establishment of reference interval values for the 13 parameters between the 3 CVs. Statistical analysis via ANOVA and Tukey's test revealed significant line differences for all 13 blood gas and chemistry parameters measured, with particularly interesting results in iCa. The blood gas and chemistry parameters collected in this study will serve as reference intervals to set the framework for potential future correlations to genetic markers, physiological abnormalities, and production performance.

Evaluation of compounded aqueous milbemycin oxime: issues with formulation potency and reproducibility.

OBJECTIVES: To determine the potency and reproducibility of milbemycin oxime when compounded as an aqueous suspension (20 mg/mL). MATERIALS AND METHODS: Preparation choice reflected current prescribing practices. Samples were acquired by prescription from two national veterinary compounding pharmacies at three time points. Two different storage conditions were evaluated and sampled at four time points from the order date (day 7, 14, 21 and 28). Milbemycin oxime recovery was performed by solid-phase extraction and concentration strength measured via high-performance liquid chromatography. RESULTS: The average concentration on day 7 for Pharmacy A samples was 16.29 mg/mL [confidence interval (CI): 15.66 to 16.92] with a coefficient of variation (CV) = 11%, while for Pharmacy B it was 20.46 mg/mL (CI: 19.83 to 21.08) with CV = 22%. The mean decrease in concentration over 28 days for Pharmacy A was 22% (CI: 9% to 34%) while Pharmacy B was 18% (CI: 2% to 35%). CLINICAL SIGNIFICANCE: The compounded milbemycin oxime suspensions evaluated in this study deviated by more than 10% from their labelled strength in five of the six lots. Clinical efficacy of compounded milbemycin oxime suspensions remains unknown and the use of these products should be discouraged at this time.

Preliminary evaluation of serum total cholesterol concentrations in dogs with osteosarcoma.

OBJECTIVES: To determine if total serum cholesterol concentrations were altered in dogs with osteosarcoma. To evaluate association of total serum cholesterol concentration with clinical outcomes in dogs with appendicular osteosarcoma. MATERIALS AND METHODS: Retrospective, multi-institutional study on 64 dogs with osteosarcoma. Control population consisted of dogs with traumatic bone fractures (n=30) and healthy patients of similar age and weight as those of the osteosarcoma cases (n=31). Survival analysis was done on 35 appendicular osteosarcoma patients that received the current standard of care. Statistical associations were assessed by univariable and multi-variable analysis. Information about age, sex, primary tumour location, total cholesterol concentration, monocytes and lymphocyte counts and alkaline phosphatase were also included. RESULTS: Total cholesterol was elevated above the reference interval (3·89 to 7·12 mmol/L) (150 to 275 mg/dL) in 29 of 64 (45·3%) osteosarcoma-bearing dogs, whereas similar elevations were found in only 3 of 30 (10%) fracture controls (P<0·0001) and 2 of 31 (6·5%) similar age/weight controls (P=0·0002). Elevated total cholesterol was significantly associated with a reduced hazard ratio (0·27, P=0·008) for overall mortality in dogs with osteosarcoma. CLINICAL SIGNIFICANCE: These results suggest that elevated total cholesterol is associated with canine osteosarcoma and may have prognostic significance.

Use of a Granulocyte Immunofluorescence Assay Designed for Humans for Detection of Antineutrophil Cytoplasmic Antibodies in Dogs with Chronic Enteropathies.

BACKGROUND: Perinuclear antineutrophil cytoplasmic antibodies (pANCA) previously have been shown to be serum markers in dogs with chronic enteropathies, with dogs that have food-responsive disease (FRD) having higher frequencies of seropositivity than dogs with steroid-responsive disease (SRD). The indirect immunofluorescence (IIF) assay used in previous publications is time-consuming to perform, with low interobserver agreement. HYPOTHESIS/OBJECTIVES: We hypothesized that a commercially available granulocyte IIF assay designed for humans could be used to detect perinuclear antineutrophil cytoplasmic antibodies in dogs. ANIMALS: Forty-four dogs with FRD, 20 dogs with SRD, 20 control dogs, and 38 soft-coated wheaten terrier (SCWT) or SCWT-cross dogs. METHODS: A granulocyte assay designed for humans was used to detect pANCA, cANCA, and antinuclear antibodies (ANA), as well as antibodies against proteinase-3 protein (PR-3) and myeloperoxidase protein (MPO) in archived serum samples. RESULTS: Sensitivity of the granulocyte assay to predict FRD in dogs was 0.61 (95% confidence interval (CI), 0.45, 0.75), and specificity was 1.00 (95% CI, 0.91, 1.00). A significant association was identified between positive pANCA or cANCA result and diagnosis of FRD (P < 0.0001). Agreement between the two assays to detect ANCA in the same serum samples from SCWT with protein-losing enteropathy/protein-losing nephropathy (PLE/PLN) was substantial (kappa, 0.77; 95% CI, 0.53, 1.00). Eight ANCA-positive cases were positive for MPO or PR-3 antibodies. CONCLUSIONS AND CLINICAL IMPORTANCE: The granulocyte immunofluorescence assay used in our pilot study was easy and quick to perform. Agreement with the previously published method was good.

Antagonism of serotonin receptor 1B decreases viability and promotes apoptosis in the COS canine osteosarcoma cell line.

Serotonin receptor 1B (5HTR1B) traditionally exhibits anti-proliferative activity in osteoblasts. We examined the expression and function of 5HTR1B in the COS canine osteosarcoma cell line and normal canine osteoblasts. Equal levels of 5HTR1B gene and protein expression were found between normal and malignant osteoblasts. Treatment with serotonin enhanced viability of osteosarcoma cells but not normal osteoblasts. Challenge with the 5HTR1B agonist anpirtoline caused no change in cell viability. Rather incubation with the specific receptor antagonist SB224289 caused reduction in osteoblast viability, with this effect more substantial in osteosarcoma cells. Investigation of this inhibitory activity showed 5HTR1B antagonism induces apoptosis in malignant cells. Evaluation of phosphorylated levels of CREB and ERK, transcriptional regulators associated with serotonin receptor signalling in osteoblasts, revealed aberrant 5HTR1B signalling in COS. Our results confirm the presence of 5HTR1B in a canine osteosarcoma cell line and highlight this receptor as a possible novel therapeutic target.