In vitro evaluation of the effect of the opioid antagonist N-methylnaltrexone on motility of the equine jejunum and pelvic flexure.

REASONS FOR PERFORMING STUDY: Although potent analgesics, opioids decrease intestinal activity, leading to ileus in many species. N-methylnaltrexone (MNTX), an opioid antagonist which does not cross the blood-brain barrier and antagonises the morphine effect on the intestine, directly stimulates motility and restores function without affecting analgesic properties. While its use has been reported in human subjects, there is no information with regard to its usage in the horse. OBJECTIVES: To determine whether MNTX has an effect on contractile activity of the equine jejunum and pelvic flexure. METHODS: Using circular smooth muscle strips obtained from 8 mature horses, increasing concentrations of MNTX were added to tissue baths in the range of 1 x 10(-9) to 1 x 10(-5) mol/l, and contractile responses were recorded for 3 mins. Data were analysed using a repeated measures ANOVA to determine whether there was a significant drug effect compared to baseline activity. Data were analysed between the jejunum and pelvic flexure using a Mann-Whitney U test. Statistical significance was established as P < 0.05. RESULTS: The administration of MNTX significantly increased the contractile frequency and amplitude at all concentrations relative to baseline (P < 0.0001) for the jejunum. The response was greatest at 1 x 10(-7) mol/l (P = 0.0005), with a mean difference from baseline of 115.12 g/cm2. The highest concentration evaluated (1 x 10(-5) mol/l) had a mean contractile strength of 69.76 g/cm2, which was significantly greater than baseline activity (P = 0.04). A significant increase in contractile activity for the colon was detected at 3 x 10(-7) mol/l and all subsequent concentrations (P < 0.04). Unlike the jejunum, the contractile activity of the pelvic flexure increased progressively with the addition of each subsequent concentration. CONCLUSIONS: N-methylnaltrexone has a direct effect on circular smooth muscle of the equine jejunum and pelvic flexure resulting in an increase in contractile activity. POTENTIAL RELEVANCE: N-methylnaltrexone could potentially be used in conjunction with morphine to provide potent and effective analgesia without compromising intestinal function. Further in vivo investigations are required to determine whether this agent antagonises morphine's effect on motility.

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.

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.

In vitro evaluation of a customized solution for use in attenuating effects of ischemia and reperfusion in the equine small intestine.

OBJECTIVE: To determine whether a customized solution could attenuate the effects of low-flow ischemia and reperfusion injury of the equine jejunum. SAMPLE POPULATION: A segment of jejunum obtained from 21 healthy adult horses. PROCEDURE: A segment of jejunum was maintained in an isolated extracorporeal circuit, and arterial flow was reduced to 20% of baseline for 40 minutes (ischemia) followed by 60 minutes of reperfusion. In 1 group, a customized solution was infused at a rate of 1 ml/min during low-flow ischemia and 3 ml/min during reperfusion. In a second group, the solution was infused at the same rate during low-flow ischemia, but it was infused at a rate of 7 ml/min during reperfusion. Control groups received lactated Ringer's solution administered at the same rates as for the customized solution. Various metabolic, hemodynamic, histologic, and permeability variables were recorded. RESULTS: A lower flow rate during reperfusion (3 ml/min) had a beneficial effect, compared with lactated Ringer's solution or the higher flow rate (7 ml/min). Use of the solution at this rate resulted in less histomorphologic injury and reduced mucosal permeability to albumin. CONCLUSIONS AND CLINICAL RELEVANCE: Use of a customized solution at a lower flow rate during repurfusion appeared to have a protective effect on equine jejunum when administered IV during low-flow ischemia and reperfusion.

The role of thermography in the management of equine lameness.

Equine thermography has increased in popularity recently because of improvements in thermal cameras and advances in image-processing software. The basic principle of thermography involves the transformation of surface heat from an object into a pictorial representation. The colour gradients generated reflect differences in the emitted heat. Variations from normal can be used to detect lameness or regions of inflammation in horses. Units can be so sensitive that flexor tendon injuries can be detected before the horse develops clinical lameness. Thermography has been used to evaluate several different clinical syndromes not only in the diagnosis of inflammation but also to monitor the progression of healing. Thermography has important applications in research for the detection of illegal performance-enhancing procedures at athletic events.

Effect of a leukocyte-depleting filter in an extracorporeal circuit used for low-flow ischemia and reperfusion of equine jejunum.

OBJECTIVE: To determine effect of leukocyte depletion on hematologic, morphologic, and metabolic variables of equine jejunum after induction of arterial low-flow ischemia and reperfusion by use of an extracorporeal circuit. ANIMALS: 14 healthy adult horses. PROCEDURE: A segment of jejunum was surgically removed and maintained in an isolated circuit for 3 hours (control group), arterial flow was reduced to 20% of baseline for 40 minutes followed by 1 hour of reperfusion (low-flow group), or leukocyte depletion was filter-induced, and low-flow ischemia and reperfusion were conducted as in the low-flow control group (filter-treated group). Various metabolic, hemodynamic, and histomorphologic variables were evaluated, including effects of electrical field stimulation and L-N-nitro-arginine-methyl-ester (L-NAME) on contractile activity. RESULTS: The extracorporeal circuit appeared to maintain the jejunum within physiologic limits for an extended period. Low-flow ischemia with reperfusion induced significant differences in various measurements, compared with control specimens. Significant differences were not detected between the low-flow and filter-treated groups. Myeloperoxidase activity was greater in the low-flow group than the control group, whereas a difference was not detected between control and filter-treated groups. CONCLUSIONS AND CLINICAL RELEVANCE: The extracorporeal circuit maintained intestine for 3 hours in a physiologic state and may be used for simulation of tissue injury. Leukocyte depletion generally did not attenuate the effects of low-flow ischemia and reperfusion on equine small intestine.

In vitro investigation of the effect of prostaglandins and nonsteroidal anti-inflammatory drugs on contractile activity of the equine smooth muscle of the dorsal colon, ventral colon, and pelvic flexure.

OBJECTIVES: To determine the in vitro effect of prostaglandin E2 (PGE2), PGF2alpha, PGI2; and nonsteroidal anti-inflammatory drugs (NSAID; ie, flunixin meglumine, ketoprofen, carprofen, and phenylbutazone) on contractile activity of the equine dorsal colon, ventral colon, and pelvic flexure circular and longitudinal smooth muscle. ANIMALS: 26 healthy horses. PROCEDURE: Tissue collected from the ventral colon, dorsal colon, and pelvic flexure was cut into strips and mounted in a tissue bath system where contractile strength was determined. Incremental doses of PGE2, PGF2alpha,, PGI2, flunixin meglumine, carprofen, ketoprofen, and phenylbutazone were added to the baths, and the contractile activity was recorded for each location and orientation of smooth muscle. RESULTS: In substance P-stimulated tissues, PGE2 and PGF2alpha enhanced contractility in the longitudinal smooth muscle with a decrease or no effect on circular smooth muscle activity. Prostaglandin I2 inhibited the circular smooth muscle response with no effect on the longitudinal muscle. The activity of NSAID was predominantly inhibitory regardless of location or muscle orientation. CONCLUSIONS AND CLINICAL RELEVANCE: In the equine large intestine, exogenous prostaglandins had a variable effect on contractile activity, depending on the location in the colon and orientation of the smooth muscle. The administration of NSAID inhibited contractility, with flunixin meglumine generally inducing the most profound inhibition relative to the other NSAID evaluated in substance P-stimulated smooth muscle of the large intestine. The results of this study indicate that prolonged use of NSAID may potentially predispose horses to develop gastrointestinal tract stasis and subsequent impaction.

Use of an extracorporeal circuit to evaluate effects of ischemia and reperfusion of the equine large colon.

OBJECTIVE: To determine efficacy of an extracorporeal circuit to maintain a segment of equine large colon for 3.5 hours and to evaluate the effect of low arterial flow on histologic and metabolic variables. SAMPLE POPULATION: Segments of large colon from 15 healthy adult horses. PROCEDURE: The pelvic flexure was surgically removed and maintained in an isolated circuit. In the control group, tissue was evaluated for 3.5 hours, whereas in the low-flow group, arterial flow was reduced to 20% of baseline for 40 minutes followed by 2 hours of reperfusion. Various metabolic and hemodynamic variables were evaluated at 30-minute intervals. Effects of nitric oxide (NO) and L-N-nitro-arginine-methyl-ester (L-NAME) on contractile activity were determined, and histomorphologic evaluation was performed at the completion of the study. RESULTS: Low-flow ischemia with reperfusion caused significant histomorphologic differences, compared with the control group. In the low-flow group, significant differences included reduction in PaCO2, reduction in bicarbonate concentrations, increase in PaO2, and an increase in base deficit in arterial and venous blood samples. Other significant differences included increases in PCV, protein concentration, total WBC count, and albumin clearance for the low-flow group. Differences were not detected in inhibitory activity of the low-flow group relative to the control tissue with or without addition of NO and L-NAME. CONCLUSION: The extracorporeal circuit maintained a segment of equine intestine for 3.5 hours and can be used to simulate ischemic injury. The extracorporeal circuit provides the potential to investigate pharmaceutic agents that can minimize intestinal injury.

Evaluation of nitric oxide as an inhibitory neurotransmitter in the equine ventral colon.

OBJECTIVE: To determine the role of nitric oxide and an apamin-sensitive nonadrenergic noncholingeric inhibitory transmitter on contractility of the ventral colon of horses. SAMPLE POPULATION: Strips of the circular and longitudinal muscle layers and taenia of the ventral colon from 14 horses. PROCEDURE: Muscle strips were suspended in tissue baths and attached to force transducers. Contractile activity of circular, longitudinal, and taenia muscle strips in response to electrical field stimulation was measured after addition of apamin and a nitric oxide inhibitor, N-nitro-L-arginine methyl ester (L-NAME). RESULTS: Electrical field stimulation reduced contractile activity in the circular muscle layer and taenia but not the longitudinal muscle layer. Addition of L-NAME significantly reduced inhibitory contractile activity at all frequencies for the circular muscle layer, whereas a significant effect was evident for the taenia only at the highest frequency. The combination of L-NAME and apamin resulted in a significant reduction in inhibition of the taenia at all frequencies but for circular muscle only at lower frequencies. CONCLUSIONS AND CLINICAL RELEVANCE: Nitric oxide and an apamin-sensitive neurotransmitter appear to mediate a component of inhibitory transmission in the circular muscle and taenia, but not the longitudinal muscle layer, of the equine ventral colon. Nitric oxide has a role in regulating contractile activity of the equine ventral colon, and nitric oxide synthase inhibitors may be useful in horses with ileus of the large colon.