Table of Contents and Abstracts

Neonatal foals may require prolonged sedation to permit ventilatory support in the first few days of life. The objective of this study was to evaluate and compare the cardiopulmonary effects and clinical recovery characteristics of 2 sedative/analgesia protocols in healthy foals receiving assisted ventilation. Foals were randomized to receive dexmedetomidine, butorphanol, and propofol (DBP) or midazolam, butorphanol, and propofol (MBP) during a 24-hour period. Infusion rates of dexmedetomidine, midazolam, and propofol were adjusted and propofol boluses administered according to set protocols to maintain optimal sedation and muscle relaxation. Ventilatory support variables were adjusted to preset targets. Physiologic variables were recorded, cardiac output (CO) measured (thermodilution), and arterial and mixed venous blood collected for gas analysis at intervals up to 24 hours. Foals in group DBP received dexmedetomidine [2.4 ± 0.5 µg/kg body weight (BW) per hour], butorphanol (13 µg/kg BW per hour), and propofol (6.97 ± 0.86 mg/kg BW per hour), whereas foals in group MBP received midazolam (0.14 ± 0.04 mg/kg BW per hour), butorphanol (13 µg/kg BW per hour), and propofol (5.98 ± 1.33 mg/kg BW per hour). Foals in the DBP group received significantly more propofol boluses (9.0 ± 3.0) than those in the MBP group (4.0 ± 2.0). Although physiologic variables remained within acceptable limits, heart rate (HR), mean arterial pressure (MAP), and cardiac index (CI) were lower in foals in the DBP group than in the MBP group. Times to sternal recumbency, standing, and nursing were significantly shorter in the DBP than MBP group. We found that MBP and DBP protocols are suitable to assist ventilatory support in neonatal foals, although MBP results in a prolonged recovery compared to DBP.

This study evaluated changes in electrocardiographic (ECG) parameters according to the stage of myxomatous mitral valve disease (MMVD) in dogs, as well as the utility of ECG parameters as prognostic indicators for congestive heart failure (CHF). Medical records of dogs with MMVD were retrospectively searched. Dogs with MMVD (N = 101) were classified into stages B [B1 (n = 52) and B2 (n = 23)] and C (n = 26) according to the American College of Veterinary Internal Medicine guidelines. Baseline variables were collected; these included signalment, radiographic, echocardiographic, and ECG parameters. Corrected QT intervals (QTc) were calculated using the logarithmic (QTc1) and Fridericia (QTc2) formulas. The P wave duration, QTc1, and QTc2 were significantly longer in stage C than in stage B. The P wave duration cutoff of 43.5 ms had a diagnostic accuracy of 65% for differentiating CHF, with a sensitivity of 63% and a specificity of 90%. A cutoff value of 307.8 ms for QTc1 yielded a sensitivity of 62%, a specificity of 76%, and a diagnostic accuracy of 78%, and a cutoff value of 239.2 ms for QTc2 yielded a sensitivity of 62%, a specificity of 83%, and a diagnostic accuracy of 77% for diagnosing CHF. Therefore, prolonged P wave and QTc in dogs with MMVD may facilitate the prediction of CHF. Electrocardiography could provide clinicians with a readily available and cost-effective screening tool for predicting CHF, if the usefulness of ECG parameters can be verified.

This study aimed to identify potential biomarkers of canine pyometra and their correlations with clinical parameters. First, 90 dogs with pyometra and 26 healthy female dogs were compared. Then, paired samples (before and after ovariohysterectomy) from 22 dogs with pyometra and 9 healthy controls from the initial cohort were compared.

Concentrations of acute inflammatory proteins, C-reactive protein (CRP) and serum amyloid A (SAA), and cell-free DNA (cfDNA), were significantly higher in dogs with pyometra than in clinically healthy dogs. Cell-free DNA was the most sensitive biomarker for systemic inflammation, based on the receiver operating characteristic curve analysis (area under the curve = 0.959). In addition, cfDNA and CRP were significantly associated with inflammation and organ injury-related clinical parameters.

Following the surgical removal of the inflamed uterus, interleukin-6 (IL-6), high-mobility group box 1 (HMGB1), and procalcitonin (PCT) significantly decreased, whereas changes in CRP, SAA, and cfDNA were not significant. These findings indicate that cfDNA, CRP, and SAA are potential clinical biomarkers of systemic inflammation in dogs with pyometra and PCT, IL-6, and HMGB1 are potential biomarkers of clinical recovery.

The objective of this retrospective study was to evaluate the expression of receptor tyrosine kinases (RTKs) in canine adrenal tumors and correlate this expression with features of tumor aggressiveness and survival in dogs undergoing adrenalectomy.

Forty-three canine adrenal tumors were evaluated for expression of c-kit, fms-like tyrosine kinase 3 (flt-3), platelet-derived growth factor receptor-β (PDGFR-β), and vascular endothelial growth factor receptor 2 (VEGFR2) using immunohistochemistry. Tumor RTK staining characteristics were compared to normal adrenals. Medical records were reviewed for data regarding patient outcome and tumor characteristics. Expression of c-kit, flt-3, PDGFR-β, and VEGFR2 was detected in 26.9%, 92.3%, 96.2%, and 61.5% of cortical tumors and 0%, 63.2%, 47.4%, and 15.8% of pheochromocytomas, respectively. Expression of RTKs was not significantly increased when compared to normal adrenals and did not correlate with survival after adrenalectomy. Receptor tyrosine kinases are not overexpressed in canine adrenal tumors compared to normal adrenal tissue. Therapeutic inhibition of these receptors may still represent an effective approach in cases where receptor activation is present.

C-reactive protein (CRP) is an acute phase protein, which is used to evaluate and monitor the response of the innate immune system to a variety of inflammatory processes in the dog. The purpose of this study was to analytically validate a point-of-care assay (IDEXX Catalyst CRP Test) and an immunoturbidimetric assay (Gentian Canine CRP Immunoassay) for the measurement of serum CRP concentrations in dogs. These 2 assays (Catalyst, Gentian) were compared to a previously validated enzyme-linked immunosorbent assay (Tridelta Development EIA Canine CRP Assay). Linearity, precision, reproducibility, and accuracy were assessed using leftover serum samples. Agreement between assays was assessed using leftover serum samples and serum from clinically healthy dogs. Observed to expected ratios (O/E) for dilutional parallelism were 83.9 to 163.1% and 108.3 to 160.6% for the Catalyst and the Gentian assays, respectively. Coefficients of variation for intra-assay variability ranged from 6.4 to 9.5% for the Catalyst assay and 1.5 to 2.6% for the Gentian assay. Coefficients of variation for inter-assay variability ranged from 3.8 to 18.2% for the Catalyst assay and 4.5 to 5.8% for the Gentian assay. The mean O/E for recovery were 97.9% and 98.5% for the Catalyst and Gentian assays, respectively. Correlations between assays were as follows: Catalyst and Tridelta (R2 = 0.76), Gentian and Tridelta (R2 = 0.79), and Catalyst and Gentian (R2 = 0.98). The Catalyst and Gentian assays are both acceptable for measuring CRP in dog serum, but their results are not directly comparable with the Tridelta assay.

Bone repair in horses implies invasive surgeries and increased cost. Research on musculoskeletal disorders therapy in horses includes cell-based therapy with mesenchymal stromal cells (MSCs). Mesenchymal stromal cells can be obtained from bone marrow (BMMSCs). Unfortunately, BMMSCs have limited cell replication in vitro. The objective of this study was to develop a biologically immortalized equine stem cell line derived from bone marrow, with unlimited in-vitro proliferation and the ability to differentiate into bone cells. Equine BMMSCs were transfected and immortalized with human telomerase reverse transcriptase (hTERT) gene. Cell passages from equine immortal BMMSCs were characterized by the presence of stemness CD markers and expression of multi-potent differentiation genes (OCT-4, SOX2, and NANOG). Equine immortal BMMSCs were incubated in osteogenic medium and bone cell differentiation was determined by alkaline phosphatase and von Kossa staining, and osteogenic gene expression (osteocalcin, Runx2, and osterix). Telomerase activity was determined by telomeric repeat amplification technique. Results showed that equine immortal BMMSCs were able to replicate in-vitro up to passage 50 and maintain stem cell characteristics by the presence of CD90 and expression of multi-potent genes. Equine immortal BMMSCs were able to differentiate into bone cells, which was confirmed by the positive osteogenic staining and gene expression. Equine BMMSCs were successfully immortalized and maintained characteristics of stem cells and readily differentiated into osteogenic cells. Extending the life span of equine BMMSCs by transfection of the hTERT gene will revolutionize the clinical use of MSCs by making them available to orthopedic surgeons “off the shelf.”

Although glucocorticoid administration has produced impressive results in treating canine mast cell tumors (MCTs), in some cases, glucocorticoids fail to reduce the tumor volume, leading to tumor relapse even after treatment. To date, mechanisms involved in glucocorticoid resistance in canine MCTs remain poorly defined. The objective of this study was to establish glucocorticoid-resistant canine MCT cell lines derived from glucocorticoid-sensitive cell lines after prolonged treatment with dexamethasone (Dex). Real-time polymerase chain reaction (RT-PCR) revealed that elevation of glucocorticoid receptor (GR)-regulated gene expression was suppressed in Dex-resistant cell lines after Dex stimulation compared with parent Dex-sensitive cell lines. This indicated that GR-regulated transcription was suppressed in Dex-resistant cell lines. Insufficient expression of GRs was not detected in Dex-resistant cell lines. Possible inhibitors of GR-regulated transcription were increased in mRNA expression in Dex-resistant cell lines. In addition, it was determined that mRNA expression of drug efflux pumps and anti-apoptosis factors was higher in Dex-resistant cell lines. In conclusion, glucocorticoid-resistant canine MCT cell lines have been established that are derived from glucocorticoid-sensitive cell lines. These cell lines suggest that multiple mechanisms contribute to glucocorticoid resistance in canine MCT cells. The mechanisms of glucocorticoid resistance after long-term treatment can be further investigated using these cell lines and a novel therapeutic strategy for glucocorticoid-resistant canine MCT cells can be developed.

Scalp electrode impedance measurements recorded by wired and wireless electroencephalography (EEG) machines in 7 healthy dogs were compared. Eight recordings resulted in 80 impedance readings from subdermal wire electrodes (locations F7/F8, F3/F4, T3/T4, C3/C4, Fz, and Cz). Impedance values were measured first from the wired and then the wireless EEG machine. Wireless impedance measurements were higher than the wired EEG machine in 79/80 readings (P ≤ 0.05), being on average 2.83 kΩ [P ≤ 0.05, 95% confidence interval (CI): 2.51 to 3.14, SD = 1.42] higher. Impedances from the wired machine ranged between < 0.5 and 9 kΩ (mean = 3.09, median = 2.00, SD = 2.15), whereas impedances from the wireless machine ranged between 2.69 and 6.07 kΩ (mean = 5.92, median = 5.05, SD = 2.59). Despite these differences in impedance measurements, both machines measured similar impedance patterns. The wireless EEG machine’s impedance measurements, therefore, should be acceptable for veterinary clinical settings.

The cross-pin technique for the treatment of distal femoral physis fractures (specifically, Salter-Harris Type I fractures) was investigated using femurs collected from beagle cadavers. The pin was inserted from the medial surface of the femur at an inclination of approximately 30 to 45° relative to the long axis of the femur in the anteroposterior direction; the pin exit was set proximal to the origin of the long digital extensor tendon. Digital and radiographic images of the femur in the anteroposterior and lateral directions were obtained. In both types of images, the insertion angle of the pin relative to the long axis was measured. Results suggest that when inserting a pin proximal to the fracture line, the ideal position can be achieved by inclining the pin approximately 20° cranially relative to the long axis of the lateral direction of the femur, in addition to the previously described criteria.