Objective: To identify suitable reference genes for normalization of real-time quantitative PCR (RT-qPCR) assay data for common tumors of dogs. Sample: Malignant lymph node (n = 8), appendicular osteosarcoma (9), and histiocytic sarcoma (12) samples and control samples of various nonneoplastic canine tissues. Procedures: Array-based comparative genomic hybridization (aCGH) data were used to guide selection of 9 candidate reference genes. Expression stability of candidate reference genes and 4 commonly used reference genes was determined for tumor samples with RT-qPCR assays and 3 software programs. Results: LOC611555 was the candidate reference gene with the highest expression stability among the 3 tumor types. Of the commonly used reference genes, expression stability of HPRT was high in histiocytic sarcoma samples, and expression stability of Ubi and RPL32 was high in osteosarcoma samples. Some of the candidate reference genes had higher expression stability than did the commonly used reference genes. Conclusions and Clinical Relevance: Data for constitutively expressed genes with high expression stability are required for normalization of RT-qPCR assay results. Without such data, accurate quantification of gene expression in tumor tissue samples is difficult. Results of the present study indicated LOC611555 may be a useful RT-qPCR assay reference gene for multiple tissue types. Some commonly used reference genes may be suitable for normalization of gene expression data for tumors of dogs, such as lymphomas, osteosarcomas, or histiocytic sarcomas.

Objective: To determine various measurements of medial retropharyngeal lymph nodes (MRPLNs) in healthy cats via ultrasonography and CT. Animals: 45 cats (age range, 2 to 8 years). Procedures: Cats underwent CT of the head and ultrasonography of the cervical region. Various measurements of MRPLNs were obtained, and parenchymal heterogeneity, presence of a hilus, appearance of margins, and attenuation of MRPLNs were determined. Results: Data for 7 cats were excluded because they did not meet inclusion criteria; data for 38 cats were evaluated. Measurements of left and right MRPLNs were not significantly different. Mean length × rostral height × rostral width dimensions of MRPLNs were 20.7 × 12.4 × 3.7 mm and 20.7 × 13.1 × 4.7 mm in ultrasonographic and CT images, respectively. Maximum MRPLN dimensions were approximately 32 × 20 × 7 mm. Mean attenuation of MRPLNs was 40.2 Hounsfield units. Parenchyma of MRPLNs was mildly (via CT) to moderately (via ultrasonography) heterogeneous. A hilus was identified in 95% (via ultrasonography) and 24% or 92% (via CT [depending on criteria used to define a hilus]) of MPRLNs. Lymph node margins were smooth in CT images and mildly irregular in ultrasonographic images. A negative linear correlation was detected between age of cat and MRPLN volume. Conclusions and Clinical Relevance: MRPLNs in cats were easily imaged via ultrasonography and CT. Left and right MRPLNs were symmetric, and MRPLNs were larger in young adult cats versus old cats. Data were intended to serve as references for evaluation of MRPLNs in healthy cats.

Objective: To determine the influence of transportation by road and air on heart rate (HR) and HR variability (HRV) in horses. Animals: 6 healthy horses. Procedures: ECG recordings were obtained from horses before (quarantine with stall rest [Q]; 24 hours) and during a journey that included transportation by road (RT; 4.5 hours), waiting on the ground in an air stall (W; 5.5 hours), and transportation by air (AT; 11 hours); HR was determined, and HRV indices of autonomic nervous activity (low-frequency [LF; 0.01 to 0.07 Hz] and high-frequency [HF; 0.07 to 0.6 Hz] power) were calculated. Results: Mean ± SD HRs during Q, RT, W, and AT were 38.9 ± 1.5 beats/min, 41.7 ± 5.6 beats/min, 41.5 ± 4.3 beats/min, and 48.8 ± 5.6 beats/min, respectively; HR during AT was significantly higher than HR during Q. The LF power was significantly higher during Q (3,454 ± 1,087 milliseconds2) and AT (3,101 ± 567 milliseconds2) than it was during RT (1,824 ± 432 milliseconds2) and W (2,072 ± 616 milliseconds2). During Q, RT, W, and AT, neither HF powers (range, 509 to 927 milliseconds2) nor LF:HF ratios (range, 4.1 to 6.2) differed significantly. The HR during RT was highly correlated with LF power (R2 = 0.979), and HR during AT was moderately correlated with the LF:HF ratio (R2 = 0.477). Conclusions and Clinical Relevance: In horses, HR and HRV indices during RT and AT differed, suggesting that exposure to different stressors results in different autonomic nervous influences on HR.

Objective: To validate the use of radiography to determine the length of sagittal otoliths in intact bogue (Boops boops; a sparid fish [commonly called sea bream]). Sample: 52 bogue cadavers. Procedures: Weight and standard lengths (from the tip of the snout to the caudal end of the last vertebra) of fish were measured. The radiographic beam was centered over the postorbital area, and images were obtained via ventrodorsal, 30° right dorsal–left ventral oblique, and 30° left dorsal–right ventral oblique projections. Otoliths were removed from the fish; sagittae were measured directly and on radiographic images by use of a vernier caliper. The relationship between direct and radiographic measurements of sagittal otoliths was calculated. Results: Paired sagittal, lapillus, and asteriscus otoliths were identified. Sagittal otoliths were comma-shaped on ventrodorsal projections; in oblique projections, they appeared ovoid and pairs of otoliths were located in the same dorsal plane. Radiographic length of sagittal otoliths was significantly correlated with directly measured length, and radiographic measurements were not significantly different between ventrodorsal and oblique radiographic projections. Conclusions and Clinical Relevance: Radiographic examination of sagittae was useful to determine the length of sagittal otoliths and, consequently, the fish length. In ecological applications, the radiographic measurement of sagittal otolith length may be useful for studies on the aquatic diet of organisms (fish, pinnipeds, and marine birds) because it allows for quick back-calculation to the size of prey.

Objective: To determine the minimum anesthetic concentration (MAC) of sevoflurane in thick-billed parrots (Rhynchopsitta pachyrhyncha) and compare MAC obtained via mechanical and electrical stimulation. Animals: 15 healthy thick-billed parrots. Procedures: Anesthesia was induced in each parrot by administration of sevoflurane in oxygen. An end-tidal sevoflurane concentration of 2.5% was established in the first bird. Fifteen minutes was allowed for equilibration. Then, 2 types of noxious stimulation (mechanical and electrical) were applied; stimuli were separated by 15 minutes. Responses to stimuli were graded as positive or negative. For a positive or negative response to a stimulus, the target end-tidal sevoflurane concentration of the subsequent bird was increased or decreased by 10%, respectively. The MAC was calculated as the mean end-tidal sevoflurane concentration during crossover events, defined as instances in which independent pairs of birds evaluated in succession had opposite responses. A quantal method was used to determine sevoflurane MAC. Physiologic variables and arterial blood gas values were also measured. Results: Via quantal analysis, mean sevoflurane MAC in thick-billed parrots determined with mechanical stimulation was 2.35% (90% fiducial interval, 1.32% to 2.66%), which differed significantly from the mean sevoflurane MAC determined with electrical stimulation, which was 4.24% (90% fiducial interval, 3.61% to 8.71%). Conclusions and Clinical Relevance: Sevoflurane MAC in thick-billed parrots determined by mechanical stimulation was similar to values determined in chickens and mammals. Sevoflurane MAC determined by electrical stimulation was significantly higher, which suggested that the 2 types of stimulation did not induce similar results in thick-billed parrots.

Objective: To evaluate 2 plate designs for pancarpal arthrodesis and their effects on load transfer to the respective bones as well as to develop a computational model with directed input from the biomechanical testing of the 2 constructs. Sample: Both forelimbs from the cadaver of an adult castrated male Golden Retriever. Procedures: CT imaging was performed on the forelimb pair. Each forelimb was subsequently instrumented with a hybrid dynamic compression plate or a castless pancarpal arthrodesis plate. Biomechanical testing was performed. The forelimbs were statically loaded in the elastic range and then cyclically loaded to failure. Finite element (FE) modeling was used to compare the 2 plate designs with respect to bone and implant stress distribution and magnitude when loaded. Results: Cyclic loading to failure elicited failure patterns similar to those observed clinically. The mean ± SD error between computational and experimental strain was < 15% ± 13% at the maximum loads applied during static elastic loading. The highest bone stresses were at the distal extent of the metacarpal bones at the level of the screw holes with both plates; however, the compression plate resulted in slightly greater stresses than did the arthrodesis plate. Both models also revealed an increase in bone stress at the proximal screw position in the radius. The highest plate stress was identified at the level of the radiocarpal bone, and an increased screw stress (junction of screw head with shaft) was identified at both the most proximal and distal ends of the plates. Conclusions and Clinical Relevance: The FE model successfully approximated the biomechanical characteristics of an ex vivo pancarpal plate construct for comparison of the effects of application of different plate designs.

Objective: To determine whether the method of lipopolysaccharide (LPS) administration (intermittent vs continuous) affects the magnitude and duration of the systemic inflammatory response in horses and whether prolonged (48 hours) endotoxemia induces laminitis. Animals: 12 healthy adult horses (10 mares and 2 geldings). Procedures: Horses were randomly assigned to receive LPS (total dose, 80 μg; n = 4) or saline (0.9% NaCl) solution (80 mL/h; 4) via constant rate infusion or 8 bolus IV injections of LPS (10 μg, q 6 h;4) during a 48-hour period. Physical examinations were performed every 4 hours, inflammatory cytokine gene expression was determined for blood samples obtained every 8 hours, and IV glucose tolerance tests were performed. Results: All LPS-treated horses had signs of depression and mild colic; those signs abated as the study progressed. Administration of LPS increased expression of interleukin-1β, interleukin-6, and interleukin-8, but results were not significantly different between LPS treatment groups. Cytokine expression was significantly higher on the first day versus the second day of LPS treatment. Interleukin-1β expression was positively correlated with rectal temperature and expression of other cytokines. Glucose and insulin dynamics for both LPS groups combined did not differ significantly from those of the saline solution group. Signs of laminitis were not detected in any of the horses. Conclusions and Clinical Relevance: Horses developed LPS tolerance within approximately 24 hours after administration was started, and the method of LPS administration did not affect the magnitude or duration of systemic inflammation. Laminitis was not induced in horses.

Objective: To determine the effects of meloxicam on values of hematologic and plasma biochemical analysis variables and results of histologic examination of tissue specimens of Japanese quail (Coturnix japonica). Animals: 30 adult Japanese quail. Procedures: 15 quail underwent laparoscopic examination of the left kidneys, and 15 quail underwent laparoscopic examination and biopsy of the left kidneys. Quail in each of these groups received meloxicam (2.0 mg/kg, IM, q 12 h; n = 10) or a saline (0.9% NaCl) solution (0.05 mL, IM, q 12 h; control birds; 5) for 14 days. A CBC and plasma biochemical analyses were performed at the start of the study and within 3 hours after the last treatment. Birds were euthanized and necropsies were performed. Results: No adverse effects of treatments were observed, and no significant changes in values of hematologic variables were detected during the study. Plasma uric acid concentrations and creatine kinase or aspartate aminotransferase activities were significantly different before versus after treatment for some groups of birds. Gross lesions identified during necropsy included lesions at renal biopsy sites and adjacent air sacs (attributed to the biopsy procedure) and pectoral muscle hemorrhage and discoloration (at sites of injection). Substantial histopathologic lesions were limited to pectoral muscle necrosis, and severity was greater for meloxicam-treated versus control birds. Conclusions and Clinical Relevance: Meloxicam (2.0 mg/kg, IM, q 12 h for 14 days) did not cause substantial alterations in function of or histopathologic findings for the kidneys of Japanese quail but did induce muscle necrosis; repeated IM administration of meloxicam to quail may be contraindicated.

Objective: To describe and measure histologic features of midcarpal joint cartilage defects in Thoroughbreds and evaluate the influence of early conditioning exercise on defect development. Sample: 24 midcarpal joints from twelve 18-month-old Thoroughbreds. Procedures: Midcarpal joints from 12 horses (6 exercised spontaneously at pasture only and 6 given additional conditioning exercise beginning at a mean age of 3 weeks were evaluated. Gross cartilage defects were assessed histologically. Third and radial carpal bones were categorized with regard to the presence or absence of calcified cartilage (CC) abnormalities at the dorsoproximal and dorsodistal articular surfaces, respectively; histomorphometric assessment and statistical analysis were conducted for the third carpal bone. Results: Number and severity of defects did not appear different between exercise groups. Nine third or radial carpal bones had thickened CC with microcracks, matrix and osteochondral junction changes, and increased vascularity, without histologic changes in the hyaline cartilage. Third carpal bones with CC abnormalities had significantly thicker CC (452 vs 228 μm) than did those without CC abnormalities in the evaluated region. However, in the same region, there were no significant differences in hyaline cartilage thickness (681 vs 603 μm), vascular channel area in the subchondral bone (624,894 vs 490,320 μm2), or number of vascular channels (15.9 vs 18.0). Conclusions and Clinical Relevance: Early exercise did not appear to influence the distribution or severity of cartilage defects in the midcarpal joint. Calcified cartilage abnormalities beneath the undisrupted hyaline cartilage in the dorsoproximal aspect of the third carpal bone may represent the first changes in the pathogenesis of midcarpal osteochondral disease.

Objective: To compare estimation of glomerular filtration rate determined via conventional methods (ie, scintigraphy and plasma clearance of technetium Tc 99m pentetate) and dynamic single-slice computed tomography (CT). Animals: 8 healthy adult cats. Procedures: Scintigraphy, plasma clearance testing, and dynamic CT were performed on each cat on the same day; order of examinations was randomized. Separate observers performed GFR calculations for scintigraphy, plasma clearance testing, or dynamic CT. Methods were compared via Bland-Altman plots and considered interchangeable and acceptable when the 95% limits of agreement (mean difference between methods ± 1.96 SD of the differences) were ≤ 0.7 mL/min/kg. Results: Global GFR differed < 0.7 mL/min/kg in 5 of 8 cats when comparing plasma clearance testing and dynamic CT; the limits of agreement were 1.4 and −1.7 mL/min/kg. The mean ± SD difference was −0.2 ± 0.8 mL/min/kg, and the maximum difference was 1.6 mL/min/kg. The mean ± SD difference (absolute value) for percentage filtration by individual kidneys was 2.4 ± 10.5% when comparing scintigraphy and dynamic CT; the maximum difference was 20%, and the limits of agreement were 18% and 23% (absolute value). Conclusions and Clinical Relevance: GFR estimation via dynamic CT exceeded the definition for acceptable clinical use, compared with results for conventional methods, which was likely attributable to sample size and preventable technical complications. Because 5 of 8 cats had comparable values between methods, further investigation of dynamic CT in a larger sample population with a wide range of GFR values should be performed.