Efficacy of single-dose intravenous phenylbutazone and flunixin meglumine before, during and after exercise in an experimental reversible model of foot lameness in horses.

REASONS FOR PERFORMING STUDY: Objective blinded efficacy data during exercise are lacking on the use of single-dose i.v. nonsteroidal anti-inflammatory drugs (NSAIDs) before, during and after exercise. HYPOTHESIS: Single i.v. doses of either phenylbutazone (PBZ) or flunixin meglumine (FM) would prove more efficacious than negative saline control (SAL) before, during and after exercise in a reversible model of foot lameness. METHODS: Six Quarter Horse mares had lameness induced by tightening a set screw against a heart bar shoe 1 h prior to treatment. Randomised blinded treatments included PBZ (4.4 mg/kg bwt i.v.), FM (1.1 mg/kg bwt i.v.), and SAL (1 ml/45 kg i.v.). Heart rate and lameness score (LS) were recorded at rest; every 20 min after lameness induction for 5 h and at the end of 2 min treadmill workloads of 2 and 4 m/s. Heart rate was also recorded from 0.5-60 min post exercise. Results were compared using RM ANOVA and Student-Newman-Keul's test (HR) and Wilcoxon signed rank test (%ΔLS) with significance set at P < 0.05. RESULTS: Pre-exercise mean HR was decreased for both NSAIDs compared to SAL from 1:20-4 h post treatment (P < 0.05). Pre-exercise mean %ΔLS was decreased for PBZ (1:20-4 h) and FM (1-4 h) compared to SAL (P < 0.01). With exercise, there were no HR differences between treatments (P > 0.05), but mean %ΔLS was decreased for both NSAIDs compared to SAL (P < 0.01). Mean recovery HR was decreased for PBZ and FM from 1-60 min compared to SAL (P < 0.05). CONCLUSIONS: PBZ and FM demonstrated definitive clinical efficacy after single i.v. doses before, during and after exercise. Use of single i.v. doses during competition may mask lameness and may affect the ability of judges in determining the soundness of horses in competition.

Forelimb skeletal scintigraphy responses in previously untrained Thoroughbreds undergoing initial treadmill training.

Scintigraphy has been used in numerous clinical settings to examine horses to determine the origin of lameness problems, but it has not been used previously to monitor prospectively the skeletal responses of a group of similarly-trained racehorses. Our hypothesis was that in naïve Thoroughbred (TB) racehorses, initial treadmill training induces increased radiopharmaceutical uptake in high-motion joints and in the dorsal third metacarpal bone (MC3). Eight previously-untrained TB racehorses underwent sequential skeletal scintigraphic examinations as they exercised daily for 9 weeks on an inclined treadmill. At the end of Weeks 0 (pre-training), 3 (trotting at 4.2 m/s and initial galloping), 6 (galloping at 7.5 m/s), and 9 (sprinting 600 m at 12.5 m/s), horses received 140 mCi 99m Technetium-methylene diphosphonate i.v. followed by a standard skeletal scintigraphic forelimb examination 2 h later. Views were graded for increased radiopharmaceutical uptake by 3 co-investigators who were blinded to horse identification, breed, sex, date, and clinical findings. Results were compared before and after training for each skeletal location using the Mann-Whitney Rank Sum Test with the level of significance set at P<0.05. Initial treadmill training resulted in increased radiopharmaceutical uptake in the carpus (P = 0.031), metacarpophalangeal joint (P = 0.021), proximal phalanx (P = 0.035), and distal phalanx (P = 0.003). Training did not affect dorsal MC3 radiopharmaceutical uptake (P>0.05).

Effects of hypochlorous acid and ascorbic acid on conductance, permeability, and structure of equine colonic mucosa in vitro.

OBJECTIVES: To study effects of hypochlorous acid (HOCl) on equine colonic mucosa in vitro, and determine whether addition of ascorbic acid protects against the effects. ANIMALS: 6 healthy horses and ponies. PROCEDURE: Short-circuit current was measured in mucosa mounted in Ussing chambers. Incubation conditions were: control (no additions); 5 mM HOCl; 1 mM HOCl; same and 5 mM ascorbic acid; 3 mM HOCl; 3 mM HOCl and 5 mM ascorbic acid; 7 mM HOCl; and 7 mM HOCl plus 5 mM ascorbic acid. Permeability was measured with [3H]mannitol and, at the conclusion of each experiment, tissues were examined microscopically to assess the effects of HOCl and ascorbic acid, alone or in combination. RESULTS: Short-circuit current and conductance increased transiently in response to 1 mM HOCl. Tissues had mild surface epithelial damage, as evident by swelling and separation of isolated cells. These changes were abolished when tissues were coincubated with 5 mM ascorbic acid and 1 mM HOCl. At 3 and 7 mM concentrations, HOCl caused marked increase in tissue conductance, short-circuit current, and permeability to mannitol; these changes were associated with histologic damage. Again, coincubation with 5 mM ascorbic acid protected against these changes. Additional studies indicated that the effects of HOCl and the protective effects of ascorbic acid were not mediated through changes in pH. CONCLUSIONS: HOCl in low concentrations is capable or increasing the short-circuit current in equine colon, possibly by increasing secretions; however, higher concentrations can cause tissue damage. The addition of 5 mM ascorbic acid blocks these changes. CLINICAL RELEVANCE: The concentration of HOCl produced by activated neutrophils could damage equine colonic mucosa and potentially contribute to or cause reperfusion injury. The ability of ascorbic acid to ameliorate this injury in an in vitro setting offers a potential method for pharmacologic evaluation of this injury and for treatment.

Effects of flunixin meglumine on short circuit current in equine colonic mucosa in vitro.

OBJECTIVES: To study the effect of flunixin meglumine on short circuit current (Isc) in equine right ventral colon in vitro. SAMPLES: Intestinal mucosa from healthy horses and ponies. PROCEDURE: Isc was measured in mucosa from the right ventral colon mounted in Ussing chambers. In experiment 1, collection and incubation solutions were: control (no additions); flunixin meglumine, 4 micrograms/ml; indomethacin, 10(-6) M; and flunixin meglumine (4 micrograms/ml) with 10(-6) M prostaglandin E2. In experiment 2, incubation conditions were: control [plain Krebs-Ringer bicarbonate [KRB] solution]; flunixin meglumine, 4 micrograms/ml in KRB; chloride-free buffer solution; flunixin meglumine (4 micrograms/ml) in a chloride-free buffer solution; and plain KRB with 10(-6) M prostaglandin E2. In experiment 3, tissue from 3 groups (n = 6 each) of animals: controls, physiologic saline solution given IV at 10 minutes before euthanasia; flunixin meglumine (1.1 mg/kg of body weight, IV) given at 10 minutes before euthanasia; and treatment similar to controls, except that tissues were incubated with 8 micrograms of flunixin meglumine/ml of bathing medium. RESULTS: Flunixin meglumine and indomethacin reduced Isc to approximately a third of control current (P < 0.05), but coincubation with flunixin meglumine and 10(-5) M prostaglandin E2 restored Isc close to the control value. Incubation with 10(-6) M prostaglandin E2 alone did not change Isc. When chloride was substituted with isethionate, flunixin meglumine had no effect on Isc. Flunixin meglumine given before euthanasia or included at a concentration of 8 micrograms/ml in all tissue preparation and incubation solutions reduced Isc (P < 0.05). CONCLUSIONS: Flunixin meglumine given IV or added to bathing solutions decreased Isc in equine right ventral colon by a mechanism that appeared to involve prostaglandin-mediated chloride secretion. CLINICAL RELEVANCE: Our results suggest that flunixin meglumine given IV to horses at recommended doses could alter putative effects of colonic prostaglandins.

Transcriptional organization of the Escherichia coli pilus adhesin K99.

The production of the Escherichia coli pilus adhesin K99 requires the expression of eight unique proteins: FanA-H. The transcriptional organization of the K99 operon was investigated by Northern blot analysis. Four RNAs of 0.54, 1.4, 2.5 and 3.5 kb were detected. When a fanC probe was used all four RNAs were detected while the use of fanD, fanF and fanG probes detected two RNAs each. Using several deletion and TnphoA insertion mutants it was concluded that there were seven unique K99-specific transcripts, several of which were of the same approximate sizes (1.4 and 2.5 kb). It also was concluded that K99 was comprised of at least three complementation groups, two of which were regulated by catabolite repression.