Objective-To evaluate the antinociceptive effects and duration of action of nalbuphine HCl administered IM on thermal thresholds in Hispaniolan Amazon parrots (Amazona ventralis). Animals-14 healthy adult Hispaniolan Amazon parrots of unknown sex. Procedures-3 doses of nalbuphine (12.5, 25, and 50 mg/kg, IM) and saline (0.9% NaCl) solution (control treatment) were evaluated in a blinded complete crossover experimental design by use of foot withdrawal threshold to a noxious thermal stimulus. Baseline data on thermal threshold were generated 1 hour before administration of nalbuphine or saline solution; thermal threshold measurements were obtained 0.5, 1.5, 3, and 6 hours after administration. Results-Nalbuphine administered IM at 12.5 mg/kg significantly increased the thermal threshold (mean change, 2.4 degrees C), compared with results for the control treatment, and significantly changed thermal threshold for up to 3 hours, compared with baseline results (mean change, 2.6 degrees to 3.8 degrees C). Higher doses of nalbuphine did not significantly change thermal thresholds, compared with results for the control treatment, but had a significant effect, compared with baseline results, for up to 3 and 1.5 hours after administration, respectively. Conclusions and Clinical Relevance-Nalbuphine administered IM at 12.5 mg/kg significantly increased the foot withdrawal threshold to a thermal noxious stimulus in Hispaniolan Amazon parrots. Higher doses of nalbuphine did not result in significantly increased thermal thresholds or a longer duration of action and would be expected to result in less analgesic effect than lower doses. Further studies are needed to fully evaluate the analgesic effects of nalbuphine in psittacine species.

Objective—To compare secretory responses to prostaglandin (PG) E2 in mucosa obtained from the proximal and distal portions of the colon of dogs. Sample—Colonic mucosa from cadavers of 18 clinically normal adult dogs. Procedures—Short-circuit current (ISC) and maximum change in ISC (ΔIsc) in response to administration of 1μM PGE2 were measured across mucosa obtained from the proximal and distal portions of the colon. Responses were evaluated in mucosa (n = 6 dogs) incubated in Ussing chambers with or without 1 mM amiloride or without chloride in the Ringer's bathing solution. Responses were also evaluated in mucosa (n = 9 dogs) incubated with or without pretreatment with 1 μM indomethacin, with or without amiloride in the subsequent bathing solution. Histologic changes in mucosa from 3 dogs were assessed over time. Results—ISC and ΔISC were significantly reduced when chloride was removed from, but not when amiloride was added to, the bathing solution and were significantly reduced after pretreatment with indomethacin. The ΔISC was significantly greater in mucosa from the distal portion of the colon than in the proximal portion of the colon. Histologic changes after incubation for 3 hours were minimal. Conclusions and Clinical Relevance—ISC and ΔISC resulted from electrogenic chloride secretion. Chloride secretion was reduced when release of PGs was prevented by indomethacin and was induced by administration of PGE2. Chloride secretion in response to PGE2 was greater in mucosa from the distal portion of the colon than in mucosa from the proximal portion of the colon.

Objective—To characterize the electroencephalogram (EEG) in young cats. Animals—23 clinically normal cats. Procedures—Cats were sedated with medetomidine hydrochloride and butorphanol tartrate at 2, 4, 6, 8, 12, 16, 20, and 24 weeks of age, and an EEG was recorded at each time point. Recordings were visually inspected for electrical continuity, interhemispheric synchrony, amplitude and frequency of background electrical activity, and frequency of transient activity. Computer-aided analysis was used to perform frequency spectral analysis and to calculate absolute and relative power of the background activity at each age. Results—Electrical continuity was evident in cats ≥ 4 weeks old, and interhemispheric synchrony was evident in cats at all ages evaluated. Analysis of amplitude of background activity and absolute power revealed significant elevations in 6-week-old cats, compared with results for 2-, 20-, and 24-week-old cats. No association between age and relative power or frequency was identified. Transient activity, which consisted of sleep spindles and K complexes, was evident at all ages, but spike and spike-and-wave discharges were observed in cats at 2 weeks of age. Conclusions and Clinical Relevance—Medetomidine and butorphanol were administered in accordance with a sedation protocol that allowed investigators to repeatedly obtain EEG data from cats. Age was an important consideration when interpreting EEG data. These data on EEG development in clinically normal cats may be used for comparison in future studies conducted to examine EEGs in young cats with diseases that affect the cerebral cortex.

Objective—To assess validity and reliability for a visual analogue scale (VAS) used by owners to measure chronic pain in their osteoarthritic dogs. Sample—68, 61, and 34 owners who completed a questionnaire. Procedures—Owners answered questionnaires at 5 time points. Criterion validity of the VAS was evaluated for all dogs in the intended-to-treat population by correlating scores for the VAS with scores for the validated Helsinki Chronic Pain Index (HCPI) and a relative quality-of-life scale. Intraclass correlation was used to assess repeatability of the pain VAS at 2 baseline evaluations. To determine sensitivity to change and face validity of the VAS, 2 blinded, randomized control groups (17 dogs receiving carprofen and 17 receiving a placebo) were analyzed over time. Results—Significant correlations existed between the VAS score and the quality-of-life scale and HCPI scores. Intraclass coefficient (r = 0.72; 95% confidence interval, 0.57 to 0.82) for the VAS indicated good repeatability. In the carprofen and placebo groups, there was poor correlation between the 2 pain evaluation methods (VAS and HCPI items) at the baseline evaluation, but the correlation improved in the carprofen group over time. No correlation was detected for the placebo group over time. Conclusions and Clinical Relevance—Although valid and reliable, the pain VAS was a poor tool for untrained owners because of poor face validity (ie, owners could not recognize their dogs' behavior as signs of pain). Only after owners had seen pain diminish and then return (after starting and discontinuing NSAID use) did the VAS have face validity.

Objective—To develop an antibody-based flow cytometric assay to detect coated platelets in dogs and to characterize the interaction of recombinant human coagulation factor VIIa with activated platelets from dogs with hemophilia A. Sample—Platelets from 4 dogs with hemophilia A, 4 dogs with hemophilia B, 4 dogs with von Willebrand disease, and 6 hemostatically normal dogs. Procedures—Freshly isolated platelets were activated with thrombin, convulxin, or a thrombin-convulxin combination. Resulting platelet phenotypes were resolved on the basis of P-selectin and fibrinogen expression, and binding of recombinant human coagulation factor VIIa to these distinct platelet subpopulations was measured by use of a flow cytometric assay. Results—Coated platelets were identified on the basis of expression of α-granule fibrinogen and were generated in response to stimulation with the thrombin-convulxin combination but not to stimulation with either agonist alone. Approximately 70% of the platelets from dogs with hemophilia A, hemophilia B, and von Willebrand disease and from the control dogs had the coated platelet phenotype. Recombinant human coagulation factor VIIa bound preferentially to coated platelets with a mean ± SD binding equilibrium constant of 2.6 ± 0.5μM. Conclusions and Clinical Relevance—Formation of coated platelets in dogs was similar to that in humans. Recombinant human coagulation factor VIIa bound preferentially to coated platelets from dogs. Impact for Human Medicine—A similar mechanism of action for recombinant human coagulation factor VIIa may exist in dogs and humans. The potential for use of dogs in the study of bleeding disorders in humans was strengthened.

This study evaluated the analgesia effects of the epidural administration of 0.1 mg/kg bodyweight (BW) of morphine or 5 micrograms/kg BW of buprenorphine in ponies with radiocarpal joint synovitis. Six ponies were submitted to 3 epidural treatments: the control group (C) received 0.15 mL/kg BW of a 0.9% sodium chloride (NaCl) solution; group M was administered 0.1 mg/kg BW of morphine; and group B was administered 5 micrograms/kg BW of buprenorphine, both diluted in 0.9% NaCl to a total volume of 0.15 mL/kg BW administered epidurally at 10 s/mL. The synovitis model was induced by injecting 0.5 ng of lipopolysaccharide (LPS) in the left or right radiocarpal joint. An epidural catheter was later introduced in the lumbosacral space and advanced up to the thoracolumbar level. The treatment started 6 h after synovitis induction. Lameness, maximum angle of carpal flexion, heart rate, systolic arterial pressure, respiratory rate, temperature, and intestinal motility were evaluated before LPS injection (baseline), 6 h after LPS injection (time 0), and 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, and 24 h after treatments. Although the model of synovitis produced clear clinical signs of inflammation, the lameness scores in group C were different from the baseline for only up to 12 h. Both morphine and buprenorphine showed a reduction in the degree of lameness starting at 0.5 and 6 h, respectively. Reduced intestinal motility was observed at 0.5 h in group M and at 0.5 to 1 h in group B. Epidural morphine was a more effective analgesic that lasted for more than 12 h and without side effects. It was concluded that morphine would be a valuable analgesic option to alleviate joint pain in the thoracic limbs in ponies.

Objective—To develop and determine the feasibility of a novel minimally invasive technique for percutaneous catheterization and embolization of the thoracic duct (PCETD) in dogs and to determine thoricic duct TD pressure at rest and during short-term balloon occlusion of the cranial vena cava (CrVC). Animals—Fifteen 7- to 11-month-old healthy mixed-breed dogs. Procedures—Efferent intestinal lymphangiography was performed, and the cisterna chyli was punctured with a trochar needle percutaneously under fluoroscopic guidance. When access was successful, a guide wire was directed into the TD through the needle and a vascular access sheath was advanced over the guide wire. Thoracic duct pressure was measured at rest and during acute balloon occlusion of the CrVC. The TD was then embolized cranial to the diaphragm with a combination of microcoils and cyanoacrylate or ethylene vinyl alcohol. Results—Successful puncture of the cisterna chyli with advancement of a wire into the TD was possible in 9 of 15 dogs, but successful catheterization was possible in only 5 of 9 dogs. Acute balloon occlusion of the CrVC led to a substantial TD pressure increase in 4 of 4 dogs, and embolization of the TD was successful in 4 of 4 dogs. Conclusions and Clinical Relevance—PCETD can successfully be performed in healthy dogs; however, this minimally invasive technique cannot currently be recommended for routine treatment of chylothorax, in part because of the technically demanding nature of the procedure. An increase in jugular venous pressure led to an increase in TD pressure, potentially predisposing some dogs to developing chylothorax.

Objective—To evaluate effects of black walnut extract (BWE) on equine mononuclear cells and determine whether BWE has direct proinflammatory effects. Sample—Mononuclear cells separated from blood samples from 8 horses. Procedures—Aqueous BWE was prepared and processed to eliminate contamination with particulates and microbes. A Limulus amoebocyte lysate assay was used to detect lipopolysaccharide (LPS) contamination in the BWE. Mononuclear cells were incubated in minimal essential medium with or without the addition of 0.6% to 10% (vol/vol) BWE. These mononuclear cells were assessed for viability, activities of caspases 3 and 7, nitric oxide production, procoagulant activity, and tumor necrosis factor-α production. The effect of LPS on cellular responses induced by BWE was assessed by coincubation with 13 U of polymyxin B/mL; mononuclear cells incubated with LPS were used as a reference. Results—BWE did not cause loss of cell membrane integrity in mononuclear cells but did induce a dose-dependent increase in activities of caspases 3 and 7. Neither BWE nor LPS significantly induced production of nitric oxide. Both BWE and LPS induced comparable amounts of procoagulant activity and tumor necrosis factor-α production; coincubation with polymyxin B reduced the activity for BWE and LPS by 50% and approximately 100%, respectively. Conclusions and Clinical Relevance—Addition of BWE induced inflammatory activation of equine mononuclear cells, a portion of which was independent of the effects of LPS. Furthermore, BWE and LPS may work in concert to induce systemic inflammatory responses that contribute to the development of acute laminitis in horses.

Objective--To evaluate 5 commercially available ELISAs for determination of leptin concentrations in serum samples from dogs. Sample Population--Serum samples from overweight-obese and thin–ideal weight clientowned dogs. Procedures--Serum samples with high and low leptin concentrations (n = 7 samples each) were used for validation of the assays. Intra- and interassay precision, linearity under dilution, spiking recovery, and limit of quantification were determined. In addition, leptin concentrations in thin–ideal weight (n = 8) and overweight-obese (37) dogs were quantified. Results--Use of 2 of the 5 ELISAs (A and B) revealed reactivity with canine leptin. Intra-and interassay coefficients of variation were < 6.1% and 76%, respectively, for assay A and 14.0% and 13.7%, respectively, for assay B. In assays A and B, dilutions of canine serum pools were used to determine linear regression equations. Recoveries were 77% to 101% for assay A and 67% to 125% for assay B. Significant differences in leptin concentrations between thin–ideal weight and overweight-obese dogs were detected only when analyzed with assay A. Conclusions and Clinical Relevance--Among 5 leptin ELISAs evaluated, a canine-specific leptin ELISA had adequate precision, linearity, and ability to discriminate between high and low leptin concentrations corresponding to overweight-obese and thin–ideal weight dogs, respectively.

Objective—To determine expression of cyclooxygenase (COX) genes 1 and 2 (also called prostaglandin-endoperoxide synthases 1 and 2) and stability of housekeeping gene expression during low-flow ischemia and reperfusion in the jejunum of horses. Animals—5 healthy adult horses. Procedures—Horses were anesthetized, and two 30-cm segments of jejunum were surgically exteriorized. Blood flow was maintained at baseline (untreated) values in 1 (control) segment and was decreased to 20% of baseline (low-flow ischemia) for 75 minutes, followed by 75 minutes of reperfusion, in the other (experimental) segment. Biopsy samples were collected from experimental segments at baseline (T0), after 75 minutes of ischemia (T1), and after 75 minutes of reperfusion (T2); samples were collected from control segments at T0 and T2. Horses were euthanized 24 hours after induction of ischemia (T3), and additional samples were collected. Samples were evaluated histologically. Total RNA was extracted; expression of COX genes and stability of 8 housekeeping genes were determined via quantitative real-time PCR assays. Results—COX-1 and COX-2 genes were constitutively expressed in baseline samples. Low-flow ischemia resulted in significant upregulation of COX-2 gene expression at each subsequent time point, compared with baseline values. The most stably expressed reference genes were β-actin and hypoxanthine phosphoribosyltransferase, whereas glyceraldehyde 3-phosphate dehydrogenase and β-2 microglobulin were the least stably expressed. Conclusions and Clinical Relevance—Low-flow ischemia resulted in upregulation of COX-2 gene expression in the jejunum of horses. Housekeeping genes traditionally used as internal standards may not be stable in this tissue during arterial low-flow ischemia and reperfusion.