The primary objective of this study was to investigate whether rider experience influences the assessment and grading of lameness in horses based on under-saddle gait analysis. Thirteen adult sports horses in active training were included in the study. After a baseline lameness and neurologic examination by the principal investigators, horses were videotaped while being ridden by an experienced and a less experienced rider. A 3-minute video was made for each horse and rider and 26 videos were randomly ordered and compiled on a DVD. Veterinarians with different levels of experience in evaluating lameness and veterinary students viewed the DVD and assigned a lameness score to each horse/rider combination. In a model accounting for the expertise of the evaluator, there was no difference in overall lameness scores between experienced and less experienced riders. This result was consistent for both sound and unsound horses. The overall lameness scores reported by specialists and students, however, differed significantly. The lameness score reported by the study participants while the horse was ridden was significantly associated with the subjective baseline lameness assessment reported by the principal investigators for the same limb when the horse was not under saddle. Additional work is necessary to determine whether riders with even lower skill levels would further alter the balance and motion pattern of the horse and have more influence on subjective grading of lameness.

Objective—To ultrasonographically measure the thickness of the individual wall layers of the duodenum, jejunum, and colon of dogs. Animals—85 dogs with no clinical signs or ultrasonographic evidence of gastrointestinal tract disease. Procedures—Total wall thickness and thickness of the mucosa, submucosa, muscularis, and serosa were measured ultrasonographically in the duodenum, jejunum, and colon of each dog. Results—The mucosal layer was the thickest layer of the duodenum and jejunum. There was a significant difference in thickness of the mucosal layer between small and large dogs. Mean ± SD thickness of the mucosal layer of the duodenum for small, medium, and large dogs was 2.4 ± 0.5 mm, 2.6 ± 0.6 mm, and 2.8 ± 0.5 mm, respectively. Mean ± SD thickness of the mucosal layer of the jejunum for small, medium, and large dogs was 1.8 ± 0.4 mm, 2.0 ± 0.4 mm, and 2.2 ± 0.5 mm, respectively. The remaining wall layers of the duodenum and jejunum were similar in thickness, and there were no significant differences among small, medium, and large dogs. All layers contributed equally to the total colonic wall thickness. Mean ± SD thickness of the colonic wall for small, medium, and large dogs was 1.5 ± 0.3 mm, 1.4 ± 0.5 mm, and 1.6 ± 0.4 mm, respectively. Conclusions and Clinical Relevance—Values for thickness of the wall layers of the duodenum, jejunum, and colon of dogs reported here may be useful for assessing gastrointestinal tract diseases primarily targeting a specific wall layer.

Objective-To determine the effects of protease inhibitors and holding times and temperatures before processing on the stability of substance P in bovine blood samples. Samples-Blood samples obtained from a healthy 6-month-old calf. Procedures-Blood samples were dispensed into tubes containing exogenous substance P and 1 of 6 degradative enzyme inhibitor treatments: heparin, EDTA, EDTA with 1 of 2 concentrations of aprotinin, or EDTA with 1 of 2 concentrations of a commercially available protease inhibitor cocktail. Plasma was harvested immediately following collection or after 1, 3, 6, 12, or 24 hours of holding at ambient (20.3° to 25.4°C) or ice bath temperatures. Total substance P immunoreactivity was determined with an ELISA; concentrations of the substance P parent molecule, a metabolite composed of the 9 terminal amino acids, and a metabolite composed of the 5 terminal amino acids were determined with liquid chromatography–tandem mass spectrometry. Results-Regarding blood samples processed immediately, no significant differences in substance P concentrations or immunoreactivity were detected among enzyme inhibitor treatments. In blood samples processed at 1 hour of holding, substance P parent molecule concentration was significantly lower for ambient temperature versus ice bath temperature holding conditions; aprotinin was the most effective inhibitor of substance P degradation at the ice bath temperature. The ELISA substance P immunoreactivity was typically lower for blood samples with heparin versus samples with other inhibitors processed at 1 hour of holding in either temperature condition. Conclusions and Clinical Relevance-Results suggested that blood samples should be chilled and plasma harvested within 1 hour after collection to prevent substance P degradation.

Objective- To describe the optimal thoracoscopic approach to the bovine pleural cavity and evaluate the short-term effects of thoracoscopy on cardiovascular and pulmonary function of healthy cattle. Sample- 6 healthy adult Holstein cows (12 hemithoraxes). Procedures- For each cow, thoracoscopy was performed in both the left and right hemithoraxes with a 24-hour interval between procedures. Cows were sedated and restrained in a standing position for each thoracoscopic examination. Examination of each hemithorax lasted for 30 minutes. Arterial blood gas variables, heart rate, and respiratory rate were assessed at predetermined times before, during, and after the procedures to monitor cardiovascular and pulmonary function. Thoracic ultrasonography was performed immediately and at 24 hours and 1 week after each thorascopic examination to evaluate the extent of residual pneumothorax. Results- Insertion of the laparoscope into the pleural cavity at the ninth intercostal space 15 cm ventral to the transverse processes of the thoracic vertebrae provided optimal visibility of structures in both the left and right hemithoraxes. Most structures of the pleural cavity were equally visible from both sides except the esophagus and the dorsal branch of the vagus nerve, which were best observed in the left hemithorax, and the pericardium, which was best observed in the right hemithorax. Mild increases in heart and respiratory rates and moderate decreases in arterial oxygen saturation and Pao2 were detected during the procedures. Conclusions and Clinical Relevance-Standing thoracoscopy was well tolerated in healthy adult dairy cattle and needs to be evaluated in cattle with pulmonary disease.

Objective-To evaluate the effects of various concentrations of l-lysine on in vitro replication of feline herpesvirus 1 (FHV-1). Sample- Cultures of Crandell-Rees feline kidney (CRFK) cells. Procedures- CRFK cells were inoculated with FHV-1 and maintained in media with 20 combinations of l-arginine and l-lysine concentrations. Changes in cell viability were monitored by continuous measurement of electrical impedance of cultured cells and by observation of viral cytopathic effects. Viral load was determined by use of quantitative PCR assay in supernatants obtained from infected cultures at specified time points. Results- Increases in l-lysine concentration had no effect on the kinetics of cell death in FHV-1-infected cultures. There was also no significant effect (r2 < 0.1) on viral DNA load for l-arginine concentrations ≥ 12 μg/mL There was a significant effect of increases in l-lysine concentration on viral DNA load in media supplemented with 6 μg of l-arginine/mL (mean ± SD slope, −4,641 ± 1,626 units; adjusted r2 = 0.45). However, the difference between the lowest (1 × 10(6.28) copies/μL) and highest (1 × 10(6.86) copies/μL) FHV-1 DNA load in these media was < 1 logarithm. Conclusions and Clinical Relevance-The difference in FHV-1 DNA load was unlikely to be biologically important. Various l-lysine concentrations did not inhibit in vitro replication of FHV-1 at l-arginine concentrations sufficient to maintain cell growth. This conclusion was consistent with results of other studies in which investigators have not detected a consistently beneficial effect when l-lysine is administered to FHV-1-infected cats.

Objective-To determine the usefulness of retina samples for detection of disease-associated prion protein by use of a commercially available enzyme immunoassay (EIA) intended for rapid identification of sheep and cattle with transmissible spongiform encephalopathies (TSEs). Samples-Retina, brainstem at the level of the obex, and retropharyngeal lymph node samples obtained from 15 TSE-inoculated sheep (scrapie [n = 13] or transmissible mink encephalopathy passaged through a bovid [2]); retina and brainstem samples obtained from 11 TSE-inoculated cattle (transmissible mink encephalopathy passaged through a bovid [7] or classical BSE [4]); and negative control tissue samples obtained from 2 sheep and 2 cattle that were not inoculated with TSEs. Procedures-Tissue samples were homogenized and analyzed for detection of abnormally folded disease-associated prion protein with a commercially available EIA and 2 confirmatory assays (western blot analysis or immunohistochemical analysis). Results-Retina sample EIA results were in agreement with results of brainstem sample EIA or confirmatory assay results for negative control animals and TSE-inoculated animals with clinical signs of disease. However, TSE-inoculated animals with positive confirmatory assay results that did not have clinical signs of disease had negative retina sample EIA results. Retina sample EIA results were in agreement with brainstem sample immunohistochemical results for 4 TSE-inoculated sheep with negative retropharyngeal lymph node EIA results. Conclusions and Clinical Relevance-Results of this study suggested that retina samples may be useful for rapid EIA screening of animals with neurologic signs to detect TSEs.

Objective—To determine the differentiation of canine adipose tissue–derived mesenchymal stem cells (ASCs) into endothelial progenitor cells (EPCs). Animals—3 healthy adult Beagles. Procedures—Canine ASCs were isolated and cultured from adipose tissue, and endothelial differentiation was induced by culturing ASCs in differentiation medium. Morphological and immunophenotypic changes were monitored. Expression of endothelial-specific markers was analyzed by conventional and real-time RT-PCR assay. The in vitro and in vivo functional characteristics of the endothelial-like cells induced from canine ASCs were evaluated by use of an in vitro solubilized basement membrane tube assay, low-density lipoprotein uptake assay, and in vivo solubilized basement membrane plug assay. Results—After differentiation culture, the cells developed morphological changes, expressed EPC markers such as CD34 and vascular endothelial growth factor receptor 2, and revealed functional characteristics in vitro. Furthermore, the induced cells allowed vessel formation in solubilized basement membrane plugs in vivo. Conclusion and Clinical Relevance—Results indicated that canine ASCs developed EPC characteristics when stimulated by differentiation medium with growth factors. Thus, differentiated canine ASC-EPCs may have the potential to provide vascularization for constructs used in regenerative medicine strategies.

Specific contrast ultrasound is widely applied in diagnostic procedures on humans but remains underused in veterinary medicine. The objective of this study was to evaluate the use of microbubble-based contrast for rapid ultrasonographic diagnosis of thrombosis in small animals, using male New Zealand white rabbits (average weight about 3.5 kg) as a model. It was hypothesized that the use of microbubble-based contrast agents will result in a faster and more precise diagnosis in our model of thrombosis. A pro-coagulant environment had been previously established by combining endothelial denudation and external vessel wall damage. Visualization of thrombi was achieved by application of contrast microbubbles [sterically stabilized, phospholipid-based microbubbles filled with sulfur hexafluoride (SF6) gas] and ultrasonography. As a result, rapid and clear diagnosis of thrombi in aorta abdominalis was achieved within 10 to 30 s (mean: 17.3 s) by applying microbubbles as an ultrasound contrast medium. In the control group, diagnosis was not possible or took 90 to 180 s. Therefore, sterically stabilized microbubbles were found to be a suitable contrast agent for the rapid diagnosis of thrombi in an experimental model in rabbits. This contrast agent could be of practical importance in small animal practice for rapid diagnosis of thrombosis.

The objective of this study was to compare the bursting strength (BS) and mode of failure (MF) of ventral midline (VM) celiotomies closed with USP 7 polydioxanone (7PD) in 1 or 2 simple continuous sections. A bursting strength model, consisting of inserting and inflating a 200-L polyurethane bladder through a 25-cm VM celiotomy, was used on 15 fresh equine cadavers. Celiotomies were closed using 7PD in 2 separate sections (4 knots), 2 continuous sections (3 knots), or a single section (2 knots) using a simple continuous pattern. The horses’ signalment, body weight, number of total knots, MF, and BS were recorded and analyzed statistically for interactions. No difference was found between the BS of VM celiotomies closure types (P = 0.4). All celiotomy/suture constructs failed at the abdominal wall. The celiotomy closure types evaluated in this study provided a secure method of closure in VM celiotomies in vivo.

Objective—To evaluate the effects of clopidogrel on clinical and clinicopathologic variables in healthy horses with experimentally induced endotoxemia. Animals—12 adult mares. Procedures—Horses were assigned with a randomization procedure to receive clopidogrel (4 mg/kg, once, then 2 mg/kg, q 24 h; n = 6) or a placebo (6) through a nasogastric tube. After 72 hours of treatment, horses received lipopolysaccharide (LPS; 30 ng/kg, IV). Heart rate, respiratory rate, rectal temperature, CBC variables, plasma fibrinogen concentration, serum tumor necrosis factor-α concentration, plasma von Willebrand factor concentration, and measures of platelet activation (including ADP- and collagen-induced platelet aggregation and closure times, thrombelastography variables, and results of flow cytometric detection of platelet membrane P-selectin, phosphatidylserine, and microparticles) were determined at various times before and after LPS administration by investigators unaware of the treatment groups. Statistical analyses were performed with repeated-measures ANOVA. Results—4 of 6 clopidogrel-treated horses had significant decreases in ADP-induced platelet aggregation before and after LPS administration. Heart rate increased significantly after LPS administration only for the placebo group. No significant differences were detected between groups for CBC variables, closure time, and plasma concentration of fibrinogen or serum concentration of tumor necrosis factor-α, and no clinically relevant differences were detected for other hemostatic variables. Conclusions and Clinical Relevance—In this study, administration of LPS did not induce platelet hyperreactivity in horses on the basis of measures of platelet adhesion, aggregation, degranulation, and procoagulant activity. Administration of clopidogrel was associated with variable platelet antiaggregatory activity and attenuated some clinical signs of endotoxemia.