Simultaneous Flow Cytometric Analysis of Phagocytosis and Oxidative Burst Activity in Equine Leukocytes

This paper describes a method for simultaneously measuring phagocytosis and oxidative burst activity in equine peripheral blood leukocytes by flow cytometry. Opsonized propidium iodide-labelled Staphylococcus aureus (PI-Sa) was used to measure the uptake of bacteria by equine phacocytes and the oxidative burst activity by oxidation of dihydrorhodamine 123. The requirements to achieve optimal activity of phagocytosis and oxidative burst are described. The advantage of the simultaneous technique is that it provides both independent and comparative values for phagocytosis and the oxidative burst, for the detection of impaired mechanisms of microbial destruction. Furthermore, the technique allows evaluation of opsonization activity in this context.     

Prognosis following surgical excision of canine cutaneous mast cell tumors with histopathologically tumor-free versus nontumor-free margins: a retrospective study of 31 cases

The purpose of this study was to determine if the presence of histopathologically tumor-free versus nontumor-free margins was prognostic for relapse or tumor-related death in dogs following surgical excision of single or multiple cutaneous mast cell tumors confined to the skin without evidence of metastasis to lymph nodes or other noncutaneous sites. Differences in tumor-related death or frequency of relapse between the two groups were not significant. Failure to achieve histopathological tumor-free margins frequently did not lead to local relapse. All tumor-related deaths occurred following local relapse. The lack of statistical support for an association between prognosis and histopathological tumor-free versus nontumor-free margins may be a result of small sample size.     

Metastatic immune infiltrates correlate with those of the primary tumour in canine osteosarcoma

Our lack of understanding of the immune microenvironment in canine osteosarcoma (cOSA) has limited the identification of potential immunotherapeutic targets. In particular, our ability to utilize readily available tissue from a dog's primary tumour to predict the type and extent of immune response in their pulmonary metastatic lesions is unknown. We, therefore, collected 21 matched pairs of primary tumours and pulmonary metastatic lesions from dogs with OSA and performed immunohistochemistry to quantify T‐lymphocyte (CD3), FOXP3+ cell, B‐lymphocyte (Pax‐5), and CD204+ macrophage infiltration. We found that T‐lymphocytes and FOXP3+ infiltrates in primary tumours positively correlated with that of metastatic lesions (ρ = 0.512, P = 0.038 and ρ = 0.698, P = 0.007, respectively), while a strong trend existed for CD204+ infiltrates (ρ = 0.404, P = 0.087). We also observed T‐ and B‐lymphocytes, and CD204+ macrophages to be significantly higher in a dog's pulmonary metastasis compared to their primary tumour (P = 0.018, P = 0.018, P = 0.016, respectively), while FOXP3+ cells were only significantly higher in metastases when all primary tumour and metastasis lesions were compared without pairing (P = 0.036). Together, these findings suggest that the metastatic immune microenvironment may be influenced by that of the primary cOSA, and that primary tumour immune biomarkers could potentially be applied to predict immunotherapeutic responses in gross metastatic disease. We, therefore, provide a rationale for the treatment of cOSA pulmonary metastases with immunotherapeutics that enhance the anti‐tumour activity of these immune cells, particularly in dogs with moderate to high immune cell infiltration in their primary tumours.     

Effects of endurance training on standard and signal-averaged electrocardiograms of sled dogs

OBJECTIVE: To determine the effect of endurance training on QRS duration, QRS-wave amplitude, and QT interval. ANIMALS: 100 sled dogs in Alaska. PROCEDURE: Dogs were examined in early September (before training) and late March (after training). During the interim, dogs trained by pulling a sled with a musher (mean, 20 km/d). Standard and signal-averaged ECG were obtained before and after training. RESULTS: Endurance training significantly increased mean QRS duration by 4.4 milliseconds for standard ECG (mean +/- SEM; 62.3 +/- 0.7 to 66.7 +/- 0.6 milliseconds) and 4.3 milliseconds for signal-averaged ECG (51.5 +/- 0.7 to 55.8 +/- 0.6 milliseconds) without changing body weight. Increase in QRS duration corresponded to a calculated increase in heart weight (standard ECG, 23%; signal-averaged ECG, 27%). Signal-averaged QRS duration was correlated with echocardiographically determined left ventricular diastolic diameter for the X orthogonal lead (r = +0.41), Y orthogonal lead (r = +0.33), and vector (r = +0.35). Training also increased QT interval (234 +/- 2 to 249 +/- 2 milliseconds) and R-wave amplitude in leads II and rV2, increased peak-to-peak voltage and S-wave amplitude in the Y orthogonal lead, and decreased Q-wave amplitude in the Y orthogonal lead. CONCLUSIONS AND CLINICAL RELEVANCE: Electrocardiographic changes reflected physiologic cardiac hypertrophy in these canine athletes in response to repetitive endurance exercise. The QRS duration increases in response to endurance exercise training and, therefore, may be of use in predicting performance in endurance activities.     

The pathology of spontaneous paratuberculosis in the North American bison (Bison bison)

Gross and histopathologic examinations were performed on 70 North American bison (Bison bison) from a Mycobacterium avium paratuberculosis culture-positive herd. The bison examined were part of a breeding herd totaling 2,800 animals. Eight of 70 (11%) animals had gross findings of intestinal mucosal thickening, and 16 of 70 (23%) of the animals had enlarged mesenteric lymph nodes. Histologic lesions compatible with Johne's disease were diagnosed in 30 of 70 (43%) bison on the basis of the demonstration of noncaseating granulomatous inflammatory infiltrates and of one or more acid-fast bacilli characteristic of Mycobacterium avium paratuberculosis. A suspicious diagnosis of Johne's disease was obtained in 11 of 70 (16%) bison on the basis of the observation of noncaseating granulomatous inflammatory infiltrates without demonstrable acid-fast bacteria. Twenty-nine of 70 (41%) animals were assessed as histologically paratuberculosis free. Histologic results were compared to Johne's disease tests such as culture, serology, and polymerase chain reaction, which were performed on some of the cohort animals.     

Plasma glucose-insulin relationship in chicken lines selected for high or low fasting glycaemia

1. Selected Fat or Lean chickens differ in their plasma glucose insulin relationship: in the fed or fasted state, Fat chickens have a lower glycaemia associated with normal or higher insulinaemia, depending upon the difference in glycaemia. 2. Conversely, chickens selected for low fasting glycaemia (LG) are fattier than their counterparts selected for high fasting glycaemia (HG), although the divergence in fat content is lower than in the Fat-Lean model. 3. The plasma glucose insulin relationship has been investigated in males of the HG and LG lines in the F4 and F5 generations. 4. A difference in glycaemia is suggested during embryonic development and was present at hatching and later on in the fasted or the fed state; insulinaemia did not differ. 5. During refeeding after an overnight fast, glycaemia differed between lines (except at intermediate times); cumulative food intake and insulinaemia were similar. 6. During a glucose tolerance test, glucose disposal rate and insulinaemia were rather similar. 7. Exogenous insulin exerted a very similar hypoglycaemic effect in both lines. 8. Other variables (body temperature, plasma concentrations of potassium and alpha NH2-non protein nitrogen) did not differ between HG and LG chickens. 9. In conclusion, HG and LG chickens do not exhibit any differences in glucose disposal rate, insulinaemia (in various nutritional conditions) or sensitivity to exogenous insulin, which contrasts with Fat or Lean chickens and may explain why HG and LG chickens have diverged to a lesser extent in fat content.     

The influence of level of feeding on growth and serum insulin-like growth factor I and insulin-like growth factor-binding proteins in growing beef cattle supplemented with somatotropin

The objective of this study was to determine the effects of level of feeding on growth, feed efficiency (gain:feed; G:F), body composition (BC), and serum concentrations of somatotropin (ST), IGF-I, and IGF-binding proteins (BP) in growing beef cattle supplemented with bovine (b) ST. In each of two consecutive years, 40 growing beef cattle were blocked by weight (average BW: yr 1 = 316 kg, yr 2 = 305 kg) and used in a 2 x 2 factorial arrangement with main effects of bST (0 or 33 microg x kg BW(-1) x d(-1)) and level of feed intake (ad libitum [AL] or 0.75 AL). Relative to uninjected cattle, treatment with bST increased ADG 9.6% (1.14 vs 1.25 kg/d; P < 0.05) and increased G:F 8.1% (12.3 vs 13.3 gain [g]:feed [kg]; P < 0.05), whereas ADG in AL animals was 39% greater than that in 0.75 AL animals (1.39 vs 1.00 kg/d; P < 0.05). There was a tendency (P = 0.10) for a bST x level of feeding interaction, such that the increase in ADG with bST was greater in AL cattle than in 0.75 AL cattle (10.6 vs 7.8%; P = 0.10). Serum concentrations of ST were greater in 0.75 AL cattle than in AL cattle (13.0 vs 8.6 ng/mL; P < 0.05) and in bST-treated cattle than in uninjected cattle (16.3 vs 5.2 ng/mL; P < 0.05). Due to a bST x level of feeding interaction (P < 0.01), the magnitude of the increase in serum ST to exogenous bST was greater (P < 0.01) in 0.75 AL cattle than in AL cattle. Relative to uninjected cattle, treatment with bST increased (P < 0.05) serum concentrations of IGF-I and IGFBP-3 and reduced (P < 0.05) concentrations of IGFBP-2. Similarly, AL cattle had greater (P < 0.05) serum concentrations of IGF-I and IGFBP-3 and reduced (P < 0.05) IGFBP-2 compared with 0.75 AL cattle. In summary, treatment with bST increased growth rate and G:F and stimulated serum IGF-I and IGFBP-3 while reducing IGFBP-2. Feeding at 0.75 ad libitum intake reduced the magnitude of response for each of these variables. Thus, limit-feeding may reduce the effect of exogenous bST on growth rate by blunting bST-induced increases in IGF-I and IGFBP-3 and bST-induced decreases in IGFBP-2.     

Evaluation of a commercially available immunoassay for assessing adequacy of passive transfer in calves

OBJECTIVE: To evaluate diagnostic utility of a commercially available immunoassay for assessing adequacy of passive transfer of immunity in neonatal calves. DESIGN: Prospective study. ANIMALS: 123 calves. PROCEDURE: Blood and serum samples were obtained from the calves prior to 2 weeks of age. The immunoassay was performed, along with refractometry and an 18% sodium sulfite turbidity test. Serum IgG concentration was determined with a radial immunodiffusion assay. Sensitivity and specificity of the immunoassay, refractometry, and the sodium sulfite test were calculated by comparing results with results of the radial immunodiffusion assay. RESULTS: Sensitivity and specificity of the blood IgG immunoassay were 0.93 and 0.88, respectively, compared with 1.00 and 0.53 for the sodium sulfite test. For refractometry, sensitivity and specificity were 0.71 and 0.83, respectively, when a serum total solids concentration of 5.2 g/dl was used as the cutoff between positive and negative test results. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the immunoassay performs well in detecting calves with inadequate passive transfer of immunity.     

Listeria associated mural and valvular endocarditis in an alpaca

An alpaca presented for emergency evaluation of respiratory distress. Physical examination revealed weakness, pyrexia, and a grade V/VI left apical holosystolic murmur. Initial thoracic examination and radiographs were consistent with left sided congestive heart failure. Blood cultures, echocardiographic evaluation, and necropsy findings established a diagnosis of bacterial endocarditis associated with a Listeria monocytogenes infection. Left ventricular endocarditis is uncommon in ruminants, and to the authors' knowledge there are no other reports of L. monocytogenes being the causative agent in an alpaca.