Objective: To obtain quantitative variables of the abdominal aorta and both kidneys on the basis of time-attenuation curves (TACs) and to measure glomerular filtration rate (GFR) for each kidney and the global GFR in clinically normal cats by use of dynamic computed tomography (CT) and Patlak analysis. Animals: 9 healthy cats. Procedures: All the cats were anesthetized with propofol. Anesthesia was maintained by administration of isoflurane, and CT examination was performed in the anesthetized cats. The TACs and renal volume were measured by use of the baseline precontrast and single-slice dynamic scans. The CT-GFR of each kidney and the global CT-GFRs were calculated via Patlak plot analysis. Results: CT-GFR results from 7 cats were valid. Peak aortic enhancement was detected between 9.0 and 14.0 seconds after iohexol injection, and the initial peak time of renal parenchymal enhancement was 15 to 24 seconds after iohexol injection. Mean ± SD global GFR was 2.06 ± 0.62 mL/min/kg. Mean ± SD CT-GFR of the right and left kidneys was 0.97 ± 0.32 mL/min/kg and 1.05 ± 0.31 mL/min/kg, respectively. Conclusions and Clinical Relevance: The CT-GFR method can be rapidly and conveniently performed in clinically normal cats. This combined structural-functional approach provided physiologic and morphological information on the kidneys of cats.

Objective—To compare the use of a semi-invasive vascular access port (VAP) device or noninvasive oscillometry versus invasive telemetry for blood pressure measurements in cats. Animals—6 healthy cats. Procedures—30 days before the study, all cats received an implanted telemeter and a VAP device. During normotension and experimentally induced hypertension, blood pressure was measured with the implanted devices and with noninvasive oscillometry at 4 time points. Results—Compared with invasive telemetry, VAP had a correlation coefficient from 0.8487 to 0.9972, and noninvasive oscillometry had a correlation coefficient from 0.7478 to 0.9689. Conclusions and Clinical Relevance—Use of the VAP device and noninvasive oscillometry had a high degree of correlation with invasive telemetry as the gold standard for blood pressure measurement. Use of a VAP device resulted in a slightly higher degree of correlation, compared with noninvasive oscillometry.

Objective—To compare the toxic effects of a Delphinium occidentale chemotype containing -(methylsuccinimido) anthranoyllycoctonine (MSAL)—type alkaloids and a D occidentale chemotype lacking MSAL-type alkaloids in mice and cattle. Animals—225 male Swiss Webster mice and 11 Black Angus steers. Procedures—4 collections of larkspur containing MSAL-type alkaloids and 4 collections of larkspur lacking MSAL-type alkaloids were used. From each collection, total alkaloid extracts (0.05 to 0.20 mL) were administered via tail-vein injection in 27 to 29 mice. Dried, finely ground plant material from 1 collection with and 1 collection without MSAL-type alkaloids (doses equivalent to 37.6 mg of total alkaloids/kg) were each administered to 8 cattle via oral gavage in a crossover experiment; 3 cattle received a single dose equivalent to 150.4 mg of total alkaloids/kg (no MSAL-type alkaloids). In mice, clinical effects were monitored; in cattle, heart rate was monitored before (baseline) and 24 hours after treatment. At the 24-hour time point, cattle were exercised as a measure of muscle weakness. Results—In mice, mean LD50 associated with alkaloid extracts prepared from plants that did or did not contain MSAL-type alkaloids was 2.3 and 54.2 mg/kg, respectively. In cattle at 24 hours after treatment, plant material containing MSAL-type alkaloids significantly increased heart rate from baseline and was associated with exercise-induced collapse; plant material lacking MSAL-type alkaloids had no similar effects. Conclusions and Clinical Relevance—Taxonomic classification of D occidentale alone was not a good indicator of the toxic risk to grazing cattle.

Objective—To evaluate whether an equine-derived canine H3N8 influenza A virus was capable of infecting and transmitting disease to ponies. Animals—20 influenza virus-seronegative 12- to 24-month-old ponies. Procedures—5 ponies were inoculated via aerosol exposure with 10(7) TCID50 of A/Canine/Wyoming/86033/07 virus (Ca/WY)/pony. A second group of 5 ponies (positive control group) was inoculated via aerosol exposure with a contemporary A/Eq/Colorado/10/07 virus (Eq/CO), and 4 sham-inoculated ponies served as a negative control group. To evaluate the potential for virus transmission, ponies (3/inoculation group) were introduced 2 days after aerosol exposure and housed with Ca/WY- and Eq/CO-inoculated ponies to serve as sentinel animals. Clinical signs, nasal virus shedding, and serologic responses to inoculation were monitored in all ponies for up to 21 days after viral inoculation. Growth and infection characteristics of viruses were examined by use of Madin-Darby canine kidney cells and primary equine and canine respiratory epithelial cells. Results—Ponies inoculated with Ca/WY had mild changes in clinical appearance, compared with results for Eq/CO-inoculated ponies. Additionally, Ca/WY inoculation induced significantly lower numbers for copies of the matrix gene in nasal secretions and lower systemic antibody responses in ponies than did Eq/CO inoculation. The Ca/WY isolate was not transmitted to sentinel ponies. Conclusions and Clinical Relevance—Inoculation of ponies with the canine H3N8 isolate resulted in mild clinical disease, minimal nasal virus shedding, and weak systemic antibody responses, compared with responses after inoculation with the equine H3N8 influenza isolate. These results suggested that Ca/WY has not maintained infectivity for ponies.

Objective--To establish a computed tomography (CT)-angiography protocol and measure the diameters of major arteries in parrots. Animals--13 Hispaniolan Amazon parrots (Amazona ventralis). Procedures--16-slice CT scanning was used to measure the apparent diameter of the ascending aorta, abdominal aorta, pulmonary arteries, and brachiocephalic trunk. Before scanning, all birds underwent ECG and echocardiographic assessment and were considered free of detectable cardiovascular diseases. Each bird was anesthetized, and a precontrast helical CT scan was performed. Peak aortic enhancement was established with a test bolus technique via dynamic axial CT scan over a predetermined single slice. An additional bolus of contrast medium was then injected, and a helical CT-angiography scan was performed immediately afterward. Arterial diameter measurements were obtained by 2 observers via various windows before and after injection, and intra- and interobserver agreement was assessed. Results--Reference limits were determined for arterial diameter measurements before and after contrast medium administration in pulmonary, mediastinal, and manual angiography windows. Ratios of vertebral body diameter to keel length were also calculated. Intraobserver agreement was high (concordance correlation coefficients ≥ 0.95); interobserver agreement was medium to high (intraclass correlation coefficients ≥ 0.65). Conclusions and Clinical Relevance--CT-angiography was safe and is of potential diagnostic value in parrots. We recommend performing the angiography immediately after IV injection of 3 mL of iohexol/kg. Arterial diameter measurements at the described locations were reliable.

Objective--To investigate the roles of transforming growth factor-β (TGF-β) isoforms and matrix metalloproteinases (MMPs) in development of chronic mitral valvular disease (CMVD) in dogs. Sample Population--12 mitral valve leaflets collected from cadavers of 5 clinically normal dogs and from 7 dogs with CMVD. Procedures--Expression of TGF-β isoforms 1, 2, and 3; MMPs 1, 2, 3, and 9; TGF-β receptor II (TβR-II); and α smooth muscle actin (αSMA) in mitral valves of dogs with CMVD was compared with that in mitral valves from clinically normal dogs. Additionally, responses of valvular interstitial cells (VICs) to TGF-β3, MMP-3, and angiotensin-converting enzyme inhibitor (ACEI) as a suppressor of TGF-β3 were examined in vitro. Results--Expression of TGF-β3, TβR-II, αSMA, and MMP-3 was only detected in mitral valves of dogs with CMVD. Concentrations of αSMA and proteoglycans in cultured VICs were significantly increased following incubation with TGF-β3; treatment with MMP-3 resulted in increased amounts of active and total TGF-β3, and total TGF-β3 in VICs was significantly decreased by incubation with ACEI. Conclusions and Clinical Relevance--Findings suggested that increased TGF-β3 and MMP-3 contribute to the pathogenesis of valvular degeneration associated with CMVD. In addition, it is possible that the use of ACEI could effectively block pathological alterations in VICs associated with CMVD in vitro. Impact on Human Medicine--CMVD is associated with primary mitral valve prolapse and Marfan syndrome in humans. Results of the study reported here will help to elucidate the molecular mechanisms of CMVD in dogs and humans.

Objective-To evaluate effects of extracorporeal shock wave therapy (ESWT) and polysulfated glycosaminoglycan treatment (PSGAGT) on subchondral bone (SCB), serum biomarkers, and synovial fluid biomarkers in horses with induced osteoarthritis. Animals-24 healthy 2- to 3-year-old horses. Procedures-An osteochondral fragment was created on the distal aspect of the radial carpal bone in 1 middle carpal joint of each horse. Horses were randomly allocated to receive local application of ESWT (days 14 and 28; n = 8), PSGAGT (IM, q 4 d for 28 days; 8), or a sham ESWT probe (placebo; days 14 and 28; 8). Serum biomarkers were measured every 7 days, and synovial fluid biomarkers were measured every 14 days. Bone density was measured by use of computed tomography on days 0 and 70, and microdamage and bone formation variables were compared among groups at the end of the study (day 70). Results-There was no significant effect of ESWT or PSGAGT on any bone variable. Serum osteocalcin concentration was significantly greater in horses that received ESWT, compared with placebo-treated horses, and serum concentration of the C-terminal telopeptide of type I collagen was significantly higher in horses that received ESWT, compared with placebo- and PSGAG-treated horses. Concentrations of the synovial fluid epitope CS846 were significantly higher in joints with osteoarthritis treated with ESWT Conclusions and Clinical Relevance-Treatment of osteoarthritis with ESWT had no effect on SCB but did induce increases in serum biomarkers indicative of bone remodeling. Treatment of osteoarthritis with PSGAG had no effect on SCB or biomarkers.

Objective-To evaluate 2 commercially available transfection reagents for transfection efficiency and distribution of small interfering RNA (siRNA) molecules to chondrocytes in monolayer cultures and full-thickness cartilage explants from guinea pigs and horses. Sample-Cartilage explants from 5 one-month-old and 3 adult guinea pigs and 5 adult clinically normal horses. Procedures-Monolayer chondrocytes and uniform cartilage explants were exposed to 1 of 2 siRNA transfection complexes according to manufacturers' protocols (1micromolar [1×]). Additionally, monolayer chondrocytes were exposed to 2× the suggested amount of a proprietary siRNA molecule. Full-thickness cartilage explants were treated with 1× (1micromolar), 2× (2micromolar), and 4× (4micromolar) or 1× (0.13micromolar), 4× (0.52micromolar), and 8× (1.04micromolar) the recommended concentrations of the proprietary siRNA and the cationic liposome siRNA, respectively, in equivalent media volumes. Use of fluorescent siRNA duplexes allowed quantification of transfected cells via flow cytometry and direct visualization of the depth and distribution of in situ transfection via fluorescent microscopy. Results-With both transfection reagents, > 90% of monolayer chondrocytes were transfected. In explants, only use of the proprietary molecule achieved > 50% transfection efficiency, whereas use of the cationic liposome achieved < 20%. Only the proprietary molecule-treated cartilage consistently contained fluorescent cells throughout all zones; the cationic liposome-transfected chondrocytes were restricted to explant surfaces. Conclusions and Clinical RelevanceRobust transfection of chondrocytes in monolayer was achieved with both reagents, but only use of the proprietary molecule attained effective full-thickness transfection of explants that may allow relevant transcript reduction via RNAi.

Objective—To evaluate cartilage thickness of the talus (especially at sites predisposed to osteochondrosis dissecans [OCD]) in growing and adult dogs not affected with OCD. Sample—Tarsocrural joints from cadavers of 34 juvenile (approx 3 months old) and 10 adult dogs. Procedures—Tarsal cartilage thickness was examined via a stereophotography microscopic system. Articular cartilage thickness was determined at 11 locations on longitudinal slices of the trochlear ridges and the sulcus between the ridges and at 2 locations in the cochlea tibiae. Cartilage thickness was measured at the proximal, proximodorsal, dorsal, and distal aspects of the trochlear ridges; proximodorsal, dorsal, and distal aspects of the trochlear sulcus; and craniolateral and caudomedial aspects of the cochlea tibiae. Differences within a joint and between sexes were evaluated. Results—Mean cartilage thickness decreased from proximal to distal in juvenile (lateral trochlear ridge, 1.52 to 0.41 mm; medial trochlear ridge, 1.10 to 0.40 mm) and from proximal to dorsal in adult (lateral trochlear ridge, 0.41 to 0.34 mm; medial trochlear ridge, 0.33 to 0.23 mm) dogs. Cartilage was thickest at the proximal aspect of the lateral trochlear ridge in both groups. Differences in proximodorsal, dorsal, and distal aspects of the ridges were not evident. Conclusions and Clinical Relevance—Healthy tarsocrural joints did not have thicker cartilage in sites predisposed to development of OCD. Evaluation of affected tarsocrural joints is necessary to exclude influences of cartilage thickness. These data are useful as a reference for distribution of cartilage thickness of the trochlea of the talus in dogs.

Objective—To develop a parsimonious statistical model to predict incidence of lameness in the subsequent lactation by use of data collected at cessation of lactation in dairy cows. Animals—574 cows. Procedures—At cessation of lactation during hoof trimming, body condition score (BCS), visual locomotion score, digital cushion thickness (DCT), and digital lesions were assessed. Results—140 (24%) cows were treated for claw horn disruption lesions (CHDLs) at cessation of lactation (114 with sole ulcers [pododermatitis circumscripta] and 26 with white line disease). The BCS was highly associated with DCT. Cows with CHDLs at cessation of lactation had significantly lower DCT, compared with other cows. All 3 logistic regression models predicted the incidence of CHDLs in the subsequent lactation with good accuracy; the area under the receiver operating characteristic curves was 0.76, 0.76, and 0.77 for the first, second, and third logistic regression models, respectively. Conclusion and Clinical Relevance—Evaluation of 3 logistic regression models indicated that lameness could be predicted with good accuracy by use of all 3. The ability to predict lameness will facilitate the implementation of lameness prevention strategies by targeting specific cows.