Up to 60% of cases of equine colitis have no known cause. To improve understanding of the causes of acute colitis in horses, we hypothesized that Clostridium perfringens producing enterotoxin (CPE) and/or beta2 toxin (CPB2) are common and important causes of severe colitis in horses and/or that C. perfringens producing an as-yet-undescribed cytotoxin may also cause colitis in horses. Fecal samples from 55 horses (43 adults, 12 foals) with clinical evidence of colitis were evaluated by culture for the presence of Clostridium difficile, C. perfringens, and Salmonella. Feces were also examined by enzyme-linked immunosorbent assay (ELISA) for C. difficile A/B toxins and C. perfringens alpha toxin (CPA), beta2 toxin (CPB2), and enterotoxin (CPE). Five C. perfringens isolates per sample were genotyped for the following genes: cpa, cpb, cpb2 consensus, cpb2 atypical, cpe (enterotoxin), etx (epsilon toxin), itx (iota toxin), netB (necrotic enteritis toxin B), and tpeL (large C. perfringens cytotoxin). The supernatants of these isolates were also evaluated for toxicity for an equine cell line. All fecal samples were negative for Salmonella. Clostridium perfringens and C. difficile were isolated from 40% and 5.4% of samples, respectively. All fecal samples were negative for CPE. Clostridium perfringens CPA and CPB2 toxins were detected in 14.5% and 7.2% of fecal samples, respectively, all of which were culture-positive for C. perfringens. No isolates were cpe, etx, netB, or tpeL gene-positive. Atypical cpb2 and consensus cpb2 genes were identified in 15 (13.6%) and 4 (3.6%) of 110 isolates, respectively. All equine C. perfringens isolates showed far milder cytotoxicity effects than a CPB-producing positive control, although cpb2-positive isolates were slightly but significantly more cytotoxic than negative isolates. Based on this studied population, we were unable to confirm our hypothesis that CPE and CPB2-producing C. perfringens are common in horses with colitis in Ontario and we failed to identify cytotoxic activity in vitro in the type A isolates recovered.

Objective—To compare repeatability and equivalency of measures of femoral trochlea depth and trochlear angle in red foxes (Vulpes vulpes) determined by use of radiography, ultrasonography, and digital photography of cadaver limbs. Sample—24 pelvic limbs from 12 red fox cadavers. Procedures—Cranioproximal-craniodistal oblique (skyline) and lateromedial radiographic views of the stifle joint and ultrasonographic images at 5 locations along the femoral trochlea were used in the study. Spacing of the 5 locations was determined on the basis of patellar position with the stifle joint at various caudal angles ranging from 96° to maximal extension (approx 170°). Ultrasonographic measurements were compared with those obtained at matched locations on photographs of anatomic preparations. Trochlear depth was assessed with all 3 image formats, and trochlear angle (measured between the trochlear ridges and sulcus) was assessed on radiographs and ultrasonographic images. Patellar thickness was measured on radiographs. Values obtained were compared by means of ANOVA, modified Bland-Altman plots, and repeatability testing. Results—Depth measurement repeatability was considered good for all modalities. Small but significant differences between mean ultrasonographic trochlear depth and anatomic (photographic) measurements were found at 3 locations; 95% limits of agreement for paired anatomic and ultrasonographic measurements were wide. The ratio of trochlear depth to radiographic patellar thickness was approximately 30% for all modalities. Trochlear angle measurements were more variable than trochlear depth measurements, especially in the distal aspect of the trochlea. Conclusions and Clinical Relevance—Paired anatomic and ultrasonographic measurements did not appear equivalent in this study, possibly attributable to imprecise probe location, which could limit quantitative use of ultrasonography in assessing proximal trochlear depth in a clinical setting.

Objective—To image the spatial distribution of pulmonary blood flow by means of scintigraphy, evaluate ventilation-perfusion (VA/Q) matching and pulmonary blood shunting (Qs/Qt) by means of the multiple inert gas elimination technique (MIGET), and measure arterial oxygenation and plasma endothelin-1 concentrations before, during, and after pulse-delivered inhaled nitric oxide (PiNO) administration to isoflurane-anesthetized horses in dorsal recumbency. Animals—3 healthy adult Standardbreds. Procedures—Nitric oxide was pulsed into the inspired gases in dorsally recumbent isoflurane-anesthetized horses. Assessment of VA/Q matching, Qs/Qt, and Pao2 content was performed by use of the MIGET, and spatial distribution of pulmonary blood flow was measured by perfusion scintigraphy following IV injection of technetium Tc 99m–labeled macroaggregated human albumin before, during, and 30 minutes after cessation of PiNO administration. Results—During PiNO administration, significant redistribution of blood flow from the dependent regions to the nondependent regions of the lungs was found and was reflected by improvements in VA/Q matching, decreases in Qs/Qt, and increases in Pao2 content, all of which reverted to baseline values at 30 minutes after PiNO administration. Conclusions and Clinical Relevance—Administration of PiNO in anesthetized dorsally recumbent horses resulted in redistribution of pulmonary blood flow from dependent atelectatic lung regions to nondependent aerated lung regions. Because hypoxemia is commonly the result of atelectasis in anesthetized dorsally recumbent horses, the addition of nitric oxide to inhaled gases could be used clinically to alleviate hypoxemia in horses during anesthesia.

Ovine pulmonary adenocarcinoma (OPA) is a transmissible lung cancer caused by Jaggsiekte sheep retrovirus (JSRV). It is difficult to identify animals infected with JSRV but are clinically healthy. The virus does not induce a specific antibody response and, although proviral DNA sequences of JSRV can be found in mononuclear blood cells, the detection is inconsistent. The aim of this study was to investigate the presence of JSRV in the bone marrow of infected sheep and develop a more consistent screening method. Immunohistochemical examination of bone marrow samples from 8 asymptomatic JSRV-infected sheep revealed the presence of positively labelled cells. However, JSRV could not be detected by a highly sensitive polymerase chain reaction (PCR) in bone marrow aspirates periodically collected from these animals. Results suggest that JSRV-infected cells may be present in the bone marrow of symptomless animals, but the number is below the detectable level for PCR. Therefore, this technique does not seem to be helpful for preclinical diagnosis of OPA.

Objective—To evaluate immunomodulatory properties of all-trans retinoic acid and a fully oxidized β-carotene dietary product in calves with Mannheimia haemolytica–induced pneumonia. Animals—Twenty-five 6- to 10-week-old male Holstein calves for experimental inoculations and three 8- to 30-week-old Angus heifers for blood donations. Procedures—In vitro, neutrophils and monocyte-derived macrophages isolated from blood of healthy Angus heifers were treated with all-trans retinoic acid (1μM) or fully oxidized β-carotene (8.3 μg/mL) for various times and assessed for markers of cellular death, antimicrobial function, and production of proinflammatory leukotriene B4. Following 28 days of dietary supplementation with fully oxidized β-carotene, Holstein calves were experimentally inoculated with M haemolytica. Bronchoalveolar lavage fluid was collected at 3 and 24 hours after challenge inoculation and analyzed for markers of apoptosis. Results—In vitro, all-trans retinoic acid and fully oxidized β-carotene induced cell-selective, caspase-3–dependent apoptosis in neutrophils, which subsequently enhanced efferocytosis in macrophages. Conversely, neither treatment altered phorbol 12-myristate 13-acetate–induced oxidative burst, phagocytosis of nonopsonized zymosan (complement or antibody independent), or M haemolytica–induced leukotriene B4 production in bovine neutrophils. In vivo, fully oxidized β-carotene enhanced leukocyte apoptosis in bronchoalveolar lavage fluid as well as subsequent efferocytosis by macrophages without altering numbers of circulating leukocytes. Conclusions and Clinical Relevance—Neutrophil apoptosis and subsequent efferocytosis by macrophages are key mechanisms in the resolution of inflammation. Findings for the present study indicated that all-trans retinoic acid and fully oxidized β-carotene could be novel nutraceutical strategies that may confer anti-inflammatory benefits for cattle with respiratory tract disease.

Objective—To determine whether stromal cell-derived factor-1 (SDF-1) concentrations in serum, plasma, and synovial fluid differed among untrained, race-trained, and osteochondral-injured Thoroughbred racehorses. Animals—22 racehorses without osteochondral injury and 37 racehorses with osteochondral injury. Procedures—Horses without osteochondral injury were examined before and after 5 to 6 months of race training. Horses with osteochondral injury were undergoing arthroscopic surgery for removal of osteochondral fragments from carpal or metacarpophalangeal or metatarsophalangeal joints (fetlock joints). Serum, plasma, and fetlock or carpal synovial fluid samples were obtained and analyzed for SDF-1 concentration by use of an ELISA. Results—In horses with fetlock or carpal joint injury, mean synovial fluid SDF-1 concentrations were significantly higher, serum SDF-1 concentrations were significantly lower, and synovial fluid-to-serum SDF-1 ratios were significantly higher than in untrained and trained horses. Synovial fluid SDF-1 concentrations were not significantly different between trained and untrained horses. Plasma SDF-1 concentrations were not different among the 3 groups. Results obtained with serum, compared with synovial fluid and plasma, had better sensitivity for differentiating between osteochondral-injured horses and uninjured horses. In horses with fetlock joint osteochondral injury, serum SDF-1 concentrations were correlated with radiographic and arthroscopic inflammation scores, but not arthroscopic cartilage scores. Conclusions and Clinical Relevance—Results suggested that serum SDF-1 concentrations were more sensitive than plasma and synovial fluid concentrations for detection of osteochondral injury in the fetlock or carpal joint of racehorses. Analysis of serum and synovial SDF-1 concentrations in horses with experimentally induced joint injury may help define the onset and progression of post-traumatic osteoarthritis and aid in the evaluation of anti-inflammatory treatments.

Objective—To develop a flow cytometric assay to quantify platelet-derived microparticles (PMPs) in equine whole blood and plasma. Sample—Citrate-anticoagulated whole blood from 30 healthy adult horses. Procedures—Platelet-poor plasma (PPP) was prepared from fresh whole blood by sequential low-speed centrifugation (twice at 2,500 × g). Samples of fresh whole blood and PPP were removed and stored at 4° and 24°C for 24 hours. Platelet-derived microparticles were characterized in fresh and stored samples on the basis of the forward scatter threshold (log forward scatter < 10(1)) and labeling with annexin V (indicating externalized phosphatidylserine) and CD61 (a constitutive platelet receptor). A fluorescent bead–calibrated flow cytometric assay was used to determine microparticle counts. Platelet counts, prothrombin time, and activated partial thromboplastin time were measured in fresh samples. Results—Significantly more PMPs were detected in fresh whole blood (median, 3,062 PMPs/μL; range, 954 to 13,531 PMPs/μL) than in fresh PPP (median, 247 PMPs/μL; range, 104 to 918 PMPs/μL). Storage at either temperature had no significant effect on PMP counts for whole blood or PPP. No significant correlation was observed between PMP counts and platelet counts in fresh whole blood or PPP or between PMP counts and clotting times in fresh PPP. Conclusions and Clinical Relevance—Results indicated that the described PMP protocol can be readily used to quantify PMPs in equine blood and plasma via flow cytometry. Quantification can be performed in fresh PPP or whole blood or samples stored refrigerated or at room temperature for 24 hours.

Objective—To compare whole-body phenylalanine kinetics and the abundance of factors in signaling pathways associated with skeletal muscle protein synthesis and protein breakdown between horses with pituitary pars intermedia dysfunction (PPID) and age-matched control horses without PPID. Animals—12 aged horses (6 horses with PPID and 6 control horses; mean age, 25.0 and 25.7 years, respectively). Procedures—Plasma glucose, insulin, and amino acids concentrations were determined before and 90 minutes after feeding. Gluteal muscle biopsy samples were obtained from horses 90 minutes after feeding, and the abundance and activation of factors involved in signaling pathways of muscle protein synthesis and breakdown were determined. The next day, horses received a priming dose and 2 hours of a constant rate infusion of 13C sodium bicarbonate followed by a priming dose and 4 hours of a constant rate infusion of 1-13C phenylalanine IV; whole-body protein synthesis was determined. Results—Plasma glucose and insulin concentrations were higher after feeding than they were before feeding for both groups of horses; however, no significant postprandial increase in plasma amino acids concentrations was detected for either group. Phenylalanine flux, oxidation, release from protein breakdown, and nonoxidative disposal were not significantly different between groups. No significant effect of PPID status was detected on the abundance or activation of positive or negative regulators of protein synthesis or positive regulators of protein breakdown. Conclusions and Clinical Relevance—Results of this study suggested that whole-body phenylalanine kinetics and the postprandial activation of signaling pathways that regulate protein synthesis and breakdown in muscles were not affected by PPID status alone in aged horses.

Objective—To determine the effects of UVB radiation produced by artificial lights on serum 25-hydroxyvitamin D concentrations in domestic rabbits (Oryctolagus cuniculi) Animals—9 juvenile domestic rabbits. Procedures—After an acclimation period, rabbits were anesthetized with isoflurane, and an initial blood sample was collected for determination of serum 25-hydroxyvitamin D concentration. Rabbits were randomly assigned to receive 12-hour exposure to UVB radiation produced by 2 compact fluorescent lights daily (n = 5) or no UVB supplementation (4) commencing on day 1. The UVB radiation emitted into the cage was measured at 9 points approximately 34 cm from the surface of the UVB light sources (representing the position of the rabbits in the cage) after 10 hours of exposure on days 1, 8, and 14. On day 14, another blood sample was collected from anesthetized rabbits for determination of serum 25-hydroxyvitamin D concentration. Results—The UVB radiation level was 8.3 to 58.1 μW/cm2 for the exposed rabbits and consistently < 0.001 μW/cm2 for the control rabbits. Mean ± SD serum 25-hydroxyvitamin D concentrations in the rabbits that were or were not provided supplemental UVB radiation for 14 days differed significantly (66.4 ± 14.3 nmol/L and 31.7 ± 9.9 nmol/L, respectively). Conclusions and Clinical Relevance—Exposure to UVB radiation produced by artificial light significantly increased serum 25-hydroxyvitamin D concentration in juvenile rabbits. Because vitamin D is an essential hormone in vertebrates, these findings suggested that the provision of supplemental UVB radiation to captive rabbits may be important.

Objective—To compare effectiveness and complications associated with peribulbar and retrobulbar anesthesia with bupivacaine in cats. Animals—6 healthy adult cats. Procedures—Cats were sedated with dexmedetomidine and received a peribulbar injection of 0.5% bupivacaine (1.5 mL), iopamidol (0.5 mL), and saline (0.9% NaCl) solution (1 mL) or retrobulbar injection of 0.5% bupivacaine (0.75 mL) and iopamidol (0.25 mL) in a crossover study with ≥ 2 weeks between treatments. The contralateral eye was the control. Injectate distribution was evaluated with CT. After atipamezole administration, periocular and corneal sensations, intraocular pressure (IOP), and ocular reflexes and appearance were evaluated for 24 hours. Results—All peribulbar and 3 of 6 retrobulbar injections resulted in CT evidence of intraconal injectate. Corneal sensation and periocular skin sensation were absent or significantly reduced relative to that for control eyes for 3 hours after peribulbar injection. Mean ± SD IOP immediately after injection was significantly higher for eyes with peribulbar injections (33 ± 12 mm Hg) than for control eyes or eyes with retrobulbar injections (both 14 ± 4 mm Hg) but 10 minutes later decreased to 18 ± 3 mm Hg. Exophthalmos, chemosis, and ptosis were evident in most injected eyes, and irritation was evident in 3 of 6 peribulbar-injected and 1 of 6 retrobulbar-injected eyes. All conditions resolved within 14 hours. Conclusions and Clinical Relevance—Peribulbar injection resulted in intraconal deposition of bupivicaine in a higher percentage of cats than did retrobulbar injection and induced notable anesthesia relative to that for the control eye; however, IOP increased temporarily.