Faye A. Harmon

Retrospective evaluation of equine prepurchase examinations performed 1991–2000

REASONS FOR PERFORMING STUDY Prepurchase examinations of horses are used increasingly as a means of evaluating future soundness. Data is lacking in the literature of the radiographic findings and results of the lameness examination of comprehensive prepurchase examinations. OBJECTIVE To summarise the clinical and radiographic findings of prepurchase examinations and determine if radiographic findings correlated with the lameness examination and sale outcome. METHODS Records of 510 cases were reviewed, radiographs evaluated and grades assigned the navicular bone, distal phalanx, and tarsus. Follow-up information on the horse status was obtained by telephone interviews for 173 horses. RESULTS Thoroughbred geldings represented the most common breed and sex, mean age 8 years, mean asking price 12,439.40 dollars, and 52.8% were lame. Radiography was the most common diagnostic procedure performed (61.6%), with views of the front feet requested most often (86.6%) followed by the tarsi (68.1%). Grade 1 was most common for the navicular bone while Grade 2 predominated for the distal phalanx. The number of sound horses decreased as grades became more severe. For the tarsi, Grades 0 and 1 were most common for the proximal intertarsal and distal intertarsal/metatarsal joints, respectively. Horses with significant tarsal changes were still able to compete at their expected level. With respect to the radiographic examination, the mean +/- s.d. grade of the horses which were not lame at follow-up was 1.2 +/- 0.9 for the navicular bone and 15 +/- 0.8 for the third phalanx. The mean +/- s.d. grade of sound horses for the distal intertarsal joint was 0.7 +/- 0.6 and 1.14 +/- 0.8 for the tarsometatarsal joint. Horses for which owner follow-up was available and which had a Grade 3 score were also evaluated. For the navicular bone, 17/31 with a Grade 3 remained in active use at follow-up and for the distal phalanx 21/27 were in active use. For the distal intertarsal and tarsometatarsal joints, 20/21 with a Grade 3 were still in active use. CONCLUSIONS Prepurchase examinations can have a significant effect on the outcome of the sale. For the navicular bone and distal phalanx, higher grades were associated with lameness. In contrast, higher grades in the tarsus were less likely to be associated with lameness. Warmbloods tended to have more extensive changes in the navicular bone and distal phalanx relative to Thoroughbreds but were not as lame. POTENTIAL RELEVANCE Radiographic changes detected in the navicular bone, distal phalanx and tarsus should be interpreted with consideration to the clinical examination.

Expression of cyclooxygenase genes in the jejunum of horses during low-flow ischemia and reperfusion.

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.

In vivo and in vitro effects of neostigmine on gastrointestinal tract motility of horses

OBJECTIVE To determine the response to neostigmine of the contractile activity of the jejunum and pelvic flexure and the effects of a continuous rate infusion (CRI) of neostigmine in horses. ANIMALS 7 adult horses and tissue from 12 adult horses. PROCEDURES A CRI of neostigmine (0.008 mg/kg/h) or placebo was administered to 6 horses in a crossover study design. Gastric emptying was evaluated by the acetaminophen test. The frequency of defecation and urination and the consistency and weight of feces were recorded throughout the experiment. The effect of neostigmine on smooth muscle contractile activity was evaluated in tissues from the jejunum and pelvic flexure. The effect of neostigmine and acetylcholine after incubation with muscarinic receptor antagonists (atropine and DAU 5884) and an acetylcholinesterase inhibitor (edrophonium) was also investigated in vitro. RESULTS No difference was observed between neostigmine and placebo for time to reach peak plasma acetaminophen concentration and absorption rate constant. A CRI of neostigmine increased fecal production and frequency of urination. Neostigmine induced a dose-dependent increase of contractile amplitude in jejunum and pelvic flexure muscle strips. Incubation of muscle strips with atropine and DAU 5884 inhibited the response to acetylcholine and neostigmine. Incubation of smooth muscle strips from the jejunum with edrophonium increased the response to acetylcholine and had no effect on the response to neostigmine in vitro. CONCLUSIONS AND CLINICAL RELEVANCE A CRI of neostigmine increased fecal production and urination frequency in horses. A CRI of neostigmine did not decrease gastric emptying. Neostigmine stimulated contractile activity of jejunum and pelvic flexure smooth muscle strips in vitro.

In vitro investigation of the interaction between nitric oxide and cyclo‐oxygenase activity in equine ventral colon smooth muscle

The objective of this study was to determine if a correlation exists between the presence of nitric oxide and prostaglandin release in the equine ventral colon smooth muscle, since this relationship may accentuate the inflammatory process during intestinal injury. Tissue was collected from the ventral colon, cut into muscle strips oriented along the circular, longitudinal and taenial layers, and mounted in a tissue bath system. Samples of the bath fluid were collected before, following electrical field stimulation (EFS), and following EFS in the presence of L-NAME, a nitric oxide synthase inhibitor. Muscle strips were also obtained following systemic administration of a cyclo-oxygnease inhibitor and samples were collected using the previously described protocol. Concentrations of prostaglandins were determined in the fluid samples using an ELISA. Electrical field stimulated release of nitric oxide produced a significant increase in prostaglandin production which did not occur in the presence of L-NAME. Systemic administration of flunixin meglumine reduced prostaglandin levels at all sampling periods, although a small increase was present following EFS. The results of this study support the hypothesis that there is a correlation between the release of nitric oxide and the production of prostaglandins in the smooth muscle of the large colon. This association between nitric oxide and prostaglandin may act as an important regulatory mechanism for various physiological mechanisms, such as vascular smooth muscle tone, and may contribute to amplified tissue injury when the induced forms of both enzymes are activated during an inflammatory insult. This suggests that the use and development of COX2 and iNOS inhibitors may help attenuate the inflammatory response following intestinal injury.