Objective—To determine the effect of short-term hyperinsulinemia on the localization and expression of endothelin receptor (ETR)-A and ETR-B in lamellar tissue of the forelimbs of horses. Samples—Distal portion of 15 cadaveric forelimbs from healthy adult horses (1 limb/horse) obtained immediately after slaughter at an abattoir. Procedures—Each forelimb was assigned to 1 of 3 treatment groups (perfused with autologous blood for 10 hours [control perfusion; n = 5], perfused with an insulin [142 ± 81 μU/mL] perfusate for 10 hours [insulinemic perfusion; 5], or not perfused [unperfused control; 5]). Immunohistochemical evaluation of lamellar tissue was performed to assess localization of ETR-A and ETR-B. Expression of ETR-A and ETR-B was measured semiquantitatively on a scale of 0 to 3 (0 = none, 1 = mild, 2 = moderate, and 3 = high-intensity staining) and quantitatively by means of gray value analysis with imaging software. Results—In all specimens, ETR-A and ETR-B were localized in endothelium, smooth muscle cells, axons, and keratinocytes. Quantitative expression of ETR-A in the midportion of the primary epidermal lamellae for the insulinemic perfusion group (149 ± 16) was lower than that for the control perfusion group (158 ± 15). Expression of ETR-B in the primary epidermal lamellae tips for the insulinemic perfusion group (140 ± 29) was higher than that for the control perfusion group (114 ± 8). Conclusions and Clinical Relevance—Hyperinsulinemia caused significant changes in endothelin receptor expression, which suggested that ETR antagonists might be beneficial for treatment of laminitis in horses.

Persistence of the oxytocin receptor (OTR) in the bovine uterus during the first 7 d after calving was investigated by means of immunohistochemical staining. Immunoreactive OTRs were present in different locations in the uterus on almost all days except day 2, when staining was seen only in the endothelium of blood vessels in the endometrium and myometrium. This finding supports the hypothesis that oxytocin may be ecbolic in cows through the 1st week post partum, but further studies are required to assess the receptor functionality during this period.

Objective—To determine the effects of protamine sulfate on clot formation time and clot strength thromboelastography variables for canine whole blood samples. Animals—Blood samples obtained from 11 healthy dogs. Procedures—Blood samples were collected from jugular veins of dogs into syringes with 3.2% sodium citrate (blood to citrate ratio, 9:1). Blood samples were divided into aliquots, and protamine sulfate was added to various concentrations (0 [control], 22, 44, and 66 μg/mL). Prepared samples were activated with kaolin (n = 8) or not activated (8), CaCl2 was added, and thromboelastography was performed. Reaction time (R), clot formation time (K), rate of clot formation (α angle), and maximum amplitude (MA) were measured. Results—For kaolin-activated and nonactivated blood samples, protamine (66 μg/mL) significantly increased R and K and decreased α angle and MA, compared with values for control samples. Also, protamine (44 μg/mL) decreased MA in nonactivated blood samples and increased K and decreased α angle in kaolin-activated samples, compared with values for control samples. Conclusions and Clinical Relevance—Results indicated protamine prolonged clot formation time and decreased overall clot strength in a dose-dependent manner; such effects may contribute to a hypocoagulable state in dogs. Kaolin-activated and nonactivated blood samples were appropriate for measurement of the effects of protamine on coagulation. Administration of protamine to reverse the effects of heparin should be performed with caution.

Objective-To develop a method for experimental induction of equine rhinitis A virus (ERAV) infection in equids and to determine the clinical characteristics of such infection. Animals-8 ponies (age, 8 to 12 months) seronegative for antibodies against ERAV. Procedures-Nebulization was used to administer ERAV (strain ERAV/ON/05; n = 4 ponies) or cell culture medium (control ponies; 4) into airways of ponies; 4 previously ERAV-inoculated ponies were reinoculated 1 year later. Physical examinations and pulmonary function testing were performed at various times for 21 days after ERAV or mock inoculation. Various types of samples were obtained for virus isolation, blood samples were obtained for serologic testing, and clinical scores were determined for various variables. Results-ERAV-inoculated ponies developed respiratory tract disease characterized by pyrexia, nasal discharge, adventitious lung sounds, and enlarged mandibular lymph nodes. Additionally, these animals had purulent mucus in lower airways up to the last evaluation time 21 days after inoculation (detected endoscopically). The virus was isolated from various samples obtained from lower and upper airways of ERAV-inoculated ponies up to 7 days after exposure; this time corresponded with an increase in serum titers of neutralizing antibodies against ERAV. None of the ponies developed clinical signs of disease after reinoculation 1 year later. Conclusions and Clinical Relevance-Results of this study indicated ERAV induced respiratory tract disease in seronegative ponies. However, ponies with neutralizing antibodies against ERAV did not develop clinical signs of disease when reinoculated with the virus. Therefore, immunization of ponies against ERAV could prevent respiratory tract disease attributable to that virus in such animals.

Objective- To evaluate the effects of tylosin on C-reactive protein concentration, carriage of Salmonella enterica, and antimicrobial resistance genes in commercial pigs. Animals-120 pigs on 2 commercial farms. Procedures- A cohort of sixty 10-week-old pigs in 4 pens/farm (15 pigs/pen) was randomly selected. Equal numbers of pigs were given feed containing tylosin (40 μg/g of feed) for 0, 6, or 12 weeks. C-reactive protein concentrations were measured, microbial culture for S enterica in feces was performed, and antimicrobial resistance genes in feces were quantified. Results- No significant associations were detected between C-reactive protein concentration or S enterica status and tylosin treatment. During the 12 weeks of tylosin administration, increased levels of 6 antimicrobial resistance genes did not occur. Conclusions and Clinical Relevance-Treatment of pigs with tylosin did not affect C-reactive protein concentration or reduce carriage or load of S enterica. There was no evidence that pigs receiving tylosin had increased carriage of the 6 antimicrobial resistance genes measured. Impact for Human Medicine-S enterica is a public health concern. Use of the antimicrobial growth promoter tylosin did not pose a public health risk by means of increased carriage of S enterica.

Objective- To assess the accuracy of an ultrasound velocity dilution cardiac output (UDCO) method, compared with that of the lithium dilution cardiac output (LiDCO) method, for determination of cardiac output (CO) in juvenile horses with experimentally induced hypovolemia. Animals- 12 anesthetized 2- to 6-month-old horses. Procedures- For each anesthetized horse, CO was determined by the LiDCO and UDCO methods prior to any intervention (baseline state), after withdrawal of approximately 40% of the horse's blood volume (low CO state), after maintenance of hypovolemia and infusion of norepinephrine until mean arterial blood pressure was equal to baseline value (high CO state), and after further infusion of norepinephrine and back-transfusion of withdrawn blood (posttransfusion state). For each of the 4 hemodynamic situations, CO and calculated cardiac index (CI) values were obtained by each method in duplicate (8 pairs of measurements/horse); mean values for each horse and overall mean values across all horses were calculated. Agreement between CI determined by each method (96 paired values) was assessed by Bland-Altman analysis. Results- For the UDCO method–derived CI measurements among the 12 horses, mean ± SD bias was −4 ± 11.3 mL/kg/min (95% limits of agreement, −26.1 to 18.2 mL/kg/min) and mean relative bias was −10.4 ± 21.5% (95% limits of agreement, −52.6% to 31.8%). Conclusions and Clinical Relevance- Results indicated that, compared with the LiDCO method, the UDCO method has acceptable clinical usefulness for determination of CO in foals.

Objective- To evaluate physical methods for inducing death during the slaughter of American alligators (Alligator mississippiensis). Animals- 24 captive hatched-and-reared American alligators. Procedures- Baseline electroencephalograms (EEGs) were obtained for awake and anesthetized alligators. Corneal reflex, spontaneous blinking, and EEGs were evaluated after severance of the spinal cord, severance of the spinal cord followed by pithing of the brain, application of a penetrating captive bolt, or application of a nonpenetrating captive bolt (6 alligators/group). Results- Overall, alligators subjected to spinal cord severance alone differed from those subjected to the other techniques. Spinal cord severance alone resulted in postprocedure EEG power values greater than those in anesthetized alligators, whereas the postprocedure EEG power values were isoelectric for the other 3 techniques. Corneal reflex and spontaneous blinking were absent in all alligators immediately after application of a penetrating or nonpenetrating captive bolt. One of 6 alligators had a corneal reflex up to 1 minute after pithing, but all others within that group had immediate cessation of reflexes after pithing. Mean time to loss of spontaneous blinking and corneal reflex for alligators subjected to spinal cord severance alone was 18 minutes (range, 2 to 37 minutes) and 54 minutes (range, 34 to 99 minutes), respectively. Conclusions and Clinical Relevance- Spinal cord severance followed by pithing of the brain and application of a penetrating or nonpenetrating captive bolt appeared to be humane and effective techniques for inducing death in American alligators, whereas spinal cord severance alone was not found to be an appropriate method.

Objective-To determine the prevalence of circumcaval ureters and other caudal vena cava variations in cats and determine whether circumcaval ureters were associated with macroscopic evidence of ureteral obstruction. Sample-301 domestic cat cadavers obtained from an animal shelter. Procedures-All cat cadavers were examined, and anatomic variations of the ureters and caudal vena cava were recorded. In cadavers with a circumcaval ureter, kidney length, width, and height were measured, and the ureters were examined macroscopically to determine whether there was gross evidence of ureteral obstruction in cats with circumcaval ureters. Results-At least 1 circumcaval ureter was present in 106 of the 301 (35.2%) cats, with a right circumcaval ureter identified in 92 (30.6%) cats, a left circumcaval ureter identified in 4 (1.3%), and bilateral circumcaval ureters identified in 10 (3.3%). Twenty-one (7.0%) cats had a double caudal vena cava, including 2 cats in which the double caudal vena cava was the only anatomic abnormality identified. No sex predilection for anatomic abnormalities was found. Mean right kidney length was significantly greater than mean left kidney length in cats with a right circumcaval ureter. Conclusions and Clinical Relevance-Circumcaval ureter was present in approximately a third of cats in this study. Variation in the development of the caudal vena cava is the proposed cause. The clinical relevance of this variation is unknown.

Objective—To investigate clinical use of proton magnetic resonance spectroscopy (1H MRS) and to compare metabolic brain bioprofiles of dogs with and without hepatic encephalopathy. Animals—6 dogs with hepatic encephalopathy and 12 control dogs. Procedures—Conventional MRI and single-voxel 1H MRS were performed with a 3-T magnet. Images for routine MRI planes and sequences were obtained. Single-voxel 1H MRS was performed with a point-resolved sequence with a short echo time (35 milliseconds) and voxel of interest placement at the level of the basal ganglia. Metabolites of interest included the glutamine-glutamate complex (sum quantification of glutamate and glutamine), myoinositol, N-acetyl aspartate, total choline, and creatine. Data were analyzed with postprocessing fitting algorithm software, and metabolite concentration relative to water and ratios with creatine as the reference metabolite were calculated. Results—Compared with control dogs, dogs with hepatic encephalopathy had specific changes, which included significantly higher concentration relative to water of the glutamine-glutamate complex and significantly lower concentration of myoinositol. Choline and N-acetyl aspartate concentrations were also slightly lower in dogs with hepatic encephalopathy than in control dogs. No differences in creatine concentration were detected between groups. Conclusions and Clinical Relevance—1H MRS aided in the diagnosis of hepatic encephalopathy in dogs, and findings supported the assumption that ammonia is a neurotoxin that manifests via glutamine-glutamate complex derangements. Use of 1H MRS may provide clinically relevant information in patients with subclinical hepatic encephalopathy, equivocal results of bile acids tests, and equivocal ammonia concentrations or may be helpful in monitoring efficacy of medical management.