Plasma and urinary concentrations of trimetoquinol by LC-MS-MS following intravenous and intra-tracheal administration to horses with heaves.

Trimetoquinol (TMQ) is a very potent and fast acting bronchodilator in horses with heaves. This study assessed the plasma and urinary concentrations of TMQ in horses with heaves following administration via the intravenous (IV, 0.2 microg/kg) and intra-tracheal (IT, 2 microg/kg) routes. TMQ was administered to six horses affected with heaves (RAO - Recurrent Airway Obstruction, used interchangeably) by the above routes and plasma and urine samples collected and stored at -20 degrees C until analyzed. Solid Phase Extraction (SPE) of TMQ was followed by highly sensitive ESI(+)-LC-MS-MS (ElectroSpray Ionization, positive mode - Liquid Chromatography - Mass Spectrometry - Mass Spectrometry); with a Limit of Detection (LOD) estimated at 1 pg/mL. Following IV administration, TMQ plasma levels peaked at 1 min at 707 pg/mL, and at 9 min at 306 pg/mL following IT administration. Our results show that TMQ plasma concentrations decline rapidly following IV administration, which is consistent with the fast onset and short duration of TMQ effect that was observed in our previous studies. On the other hand, IT administration showed a very unique plasma concentration pattern. From a regulatory standpoint, the current available TMQ ELISA kit was also used in an attempt to detect TMQ from the plasma and urine samples. We report that the ELISA kit was unable to detect TMQ from any of the samples generated in these studies.

Pulmonary response to airway instillation of autologous blood in horses.

REASONS FOR PERFORMING STUDY: Exercise-induced pulmonary haemorrhage (EIPH) occurs in the majority of horses performing strenuous exercise. Associated pulmonary lesions include alveolar and airway wall fibrosis, which may enhance the severity of EIPH. Further work is required to understand the pulmonary response to blood in the equine airways. OBJECTIVES: To confirm that a single instillation of autologous blood into horse airways is associated with alveolar wall fibrosis, and to determine if blood in the airways is also associated with peribronchiolar fibrosis. METHODS: Paired regions of each lung were inoculated with blood or saline at 14 and 7 days, and 48, 24 and 6 h before euthanasia. Resulting lesions were described histologically and alveolar and airway wall collagen was quantified. RESULTS: The main lesion observed on histology was hypertrophy and hyperplasia of type II pneumocytes at 7 days after blood instillation. This lesion was no longer present at 14 days. There were no significant effects of lung region, treatment (saline or autologous blood instillation), nor significant treatment-time interactions in the amount of collagen in the interstitium or in the peribronchial regions. CONCLUSION: A single instillation of autologous blood in lung regions is not associated with pulmonary fibrosis. POTENTIAL RELEVANCE: Pulmonary fibrosis and lung remodelling, characteristic of EIPH, are important because these lesions may enhance the severity of bleeding during exercise. A single instillation of autologous blood in the airspaces of the lung is not associated with pulmonary fibrosis. Therefore the pulmonary fibrosis described in EIPH must have other causes, such as repetitive bleeds, or the presence of blood in the pulmonary interstitium in addition to the airspaces. Prevention of pulmonary fibrosis through therapeutic intervention requires a better understanding of these mechanisms.

Trimetoquinol: bronchodilator effects in horses with heaves following aerosolised and oral administration.

REASON FOR PERFORMING STUDY: The bronchodilator effects of trimetoquinol (TMQ) have been studied when administered i.v. or intratracheally, but not in an aerosolised form. OBJECTIVES: To define the relationship between the therapeutic and adverse responses (therapeutic index) of TMQ when administered as an aerosol or by the oral route. METHODS: Increasing doses of TMQ were administered to horses with heaves as an aerosol and by the oral route. Dose ranged 100-1000 microg/horse for aerosolised TMQ and from 6-60 microg/kg bwt for the oral route. Airway and cardiac effects were assessed by measurement of maximal change in pleural pressure (deltaPplmax) and heart rate (HR), respectively. Side effects of sweating, agitation and muscle trembling were scored subjectively. Duration of action of aerosolised (1000 pg/horse) and oral (6-60 microg/kg bwt) TMQ was evaluated over 6 h. RESULTS: Aerosol administration of TMQ caused dose-dependent bronchodilation but did not change HR or cause other observable side effects. When 1000 microg/horse was administered via aerosol, TMQ produced a 2-phase bronchodilation; an immediate effect lasting up to 30 min and a second phase between 2 and 4 h. Oral TMQ was therapeutically ineffective. CONCLUSION: Aerosol administration of TMQ is a safe and effective method of producing bronchodilation in horses.

Intravenous and intratracheal administration of trimetoquinol, a fast-acting short-lived bronchodilator in horses with 'heaves'.

REASON FOR PERFORMING STUDY: Trimetoquinol (TMQ) is a potent beta-adrenoceptor agonist bronchodilator used in human medicine but has not been evaluated for potential use as a therapeutic agent for horses with 'heaves'. OBJECTIVES: To assess the pharmacodynamics of TMQ in horses with 'heaves' to determine potential therapeutic effects. METHODS: Increasing doses of TMQ were administered to horses with 'heaves' by i.v. and intratracheal (i.t.) routes. Doses ranged 0.001-0.2 microg/kg bwt i.v. and 0.01-2 microg/kg bwt i.t. Cardiac and airways effects were assessed by measurement of heart rate (HR) and maximal change in pleural pressure (deltaPplmax), respectively. Side effects of sweating, agitation and muscle trembling were scored subjectively. Duration of action to i.v. (0.2 microg/kg bwt) and i.t. (2 microg/kg bwt) TMQ was evaluated over 6 h. RESULTS: Intravenous TMQ was an exceptionally potent cardiac stimulant. Heart rate increased at 0.01 microg/kg bwt, and was still increasing after administration of highest dose, 0.2 microg/kg bwt. Airway bronchodilation, measured as a decrease in deltaPplmax, also commenced at 0.01 microg/kg bwt. By the i.t. route, TMQ was 50-100-fold less potent than by i.v. Side effects included sweating, agitation and muscle trembling. Overall, the onset of HR and bronchodilator effects was rapid, within about 3 min, but effects were over at 2 h. CONCLUSION: When administered i.v. and i.t., TMQ is a highly potent cardiac stimulant and a modest bronchodilator. It may not be an appropriate pharmacological agent by i.v. and i.t. routes for the alleviation of signs in horses with 'heaves'. Further studies of TMQ by oral and aerosol routes are necessary. POTENTIAL RELEVANCE: In horses, TMQ is a fast-acting bronchodilator with a short duration of action. It could be used as a rescue agent during an episode of 'heaves'. The i.v. and i.t. administration of TMQ is associated with side effects, similar to those reported for all other beta-agonists. However, other routes, such as aerosol and oral, may prove useful and safe for the alleviation of bronchoconstriction typical of 'heaves'.

Effect of tracheal mucus and tracheal cytology on racing performance in Thoroughbred racehorses.

REASON FOR PERFORMING STUDY: Accumulations of mucus within the trachea are often found during endoscopic examinations of the airways of poorly performing racehorses, but the clinical importance of this finding is unknown. OBJECTIVES: To determine the effect of tracheal mucus, pharyngeal lymphoid hyperplasia (PLH) and cytological indices of tracheal aspirate on racing performance in Thoroughbred horses assessed by race place and whether the horse was raced. METHODS: Endoscopic examination of the nasopharynx, larynx and trachea was performed, and a tracheal aspirate obtained monthly at Thistledown racetrack from April to December, 2002 and 2003. Horses received a score of 0-4 for the degree of PLH and 0-4 for the amount of mucus visible in the trachea. The tracheal aspirate was assessed for turbidity, and total and differential cell counts. Generalised estimating equations models were used as repeated measures models for each risk factor and the level of association assessed through the risk factor's P value in the model. RESULTS: Moderate to severe tracheal mucus (2-4) was a risk factor for poor racing performance. There was no association between degree of PLH, cell counts or turbidity of tracheal wash fluid and racing performance. However, horses that raced had higher total neutrophil counts in tracheal wash aspirates than horses that did not race. CONCLUSIONS: Grades 2-4 tracheal mucus should be considered a potential cause of poor racing performance in Thoroughbred horses. CLINICAL RELEVANCE: Because moderate to severe tracheal mucus accumulation, and not increased tracheal neutrophils, was a risk factor for poor racing performance, functionally significant airway inflammation may best be confirmed by the presence of mucus rather than increased number of neutrophils in the trachea.

Response to nasopharyngeal oxygen administration in horses with lung disease.

REASONS FOR PERFORMING STUDY: Guidelines for administration of oxygen to standing horses are unavailable because previous investigations of the efficacy of oxygen administration to increase arterial oxygenation in standing horses have produced equivocal results. OBJECTIVE: To determine the effect of nasal oxygen supplementation on inspired and arterial blood gas tensions in control horses and those with moderate to severe recurrent airway obstruction (RAO). METHODS: Normal horses (n = 6) and horses during an attack of RAO induced by stabling (n = 6) were studied. Oxygen was administered through either one or 2 cannulae, passed via the nares into the nasopharynx to the level of the medial canthus of each eye. Intratracheal inspired oxygen and carbon dioxide concentration and arterial blood gas tensions were measured at baseline and during delivery of 5, 10, 15, 20 and 30 l/min oxygen. RESULTS: Nasal cannulae and all but the highest oxygen flow rates were well tolerated. Fractional inspired oxygen concentration (F(I)O2) increased with flow but was significantly lower at all flow rates in horses with RAO compared with controls. Arterial oxygen tension (PaO2) was significantly increased (P < 0.001) by all flow rates, but was always lower in RAO-affected animals. At 30 l/min, PaO2 increased to 319 +/- 31 mmHg in control horses and 264 +/- 69 mmHg in horses with RAO. Additionally, a large arterial to end-tidal gradient for CO2 in RAO-affected horses was observed, indicating increased alveolar deadspace ventilation in these animals. CONCLUSIONS: The use of nasal cannulae to deliver oxygen effectively increases both F(I)O2 and PaO2 in horses with moderate to severe RAO. Oxygen flow rates up to 20 l/min are well tolerated, but flow rates of 30 l/min produce occasional coughing or gagging. POTENTIAL RELEVANCE: Oxygen therapy delivered by means of an intranasal cannula is a highly effective means of increasing arterial oxygen tension in horses with respiratory disease. Generally, flows of 10-20 l/min should be effective. If higher flows (20-30 l/min) are necessary, they should be delivered by means of 2 cannulae.

Thermographic study of in vivo modulation of vascular responses to phenylephrine and endothelin-1 by dexamethasone in the horse.

REASONS FOR PERFORMING STUDY: In vitro, glucocorticoids potentiate vasoconstriction of equine digital vessels to catecholamines and this has been implicated as a mechanism of glucocorticoid-induced laminitis. This observation has never been confirmed in vivo. OBJECTIVES: To study the effects of glucocorticoid therapy on vasoconstrictor responsiveness in the horse in vivo. METHODS: In a blinded, randomised cross-over experiment, 9 horses were treated with either dexamethasone (0.1 mg/kg bwt i.v. q. 24 h) or saline i.v. for 6 days. The changes in local average skin temperature before (baseline) and after intradermal injections of the alpha1-adrenoceptor agonist phenylephrine (PHE; 10(-4), 10(-5), 10(-6), 10(-7) and 10(-8) mol/l), endothelin-1 (ET-1; 10(-5), 10(-6), 10(-7), 10(-8) and 10(-9) mol/l) or ET-1 plus a blocker (BQ-123 10(-6) mol/l; RES-701 10(-6) mol/l; and L-NAME 10(-4) mol/l) were investigated with a thermograph. RESULTS: Dexamethasone (DEX) decreased baseline skin temperatures, suggesting reduced blood flow as a consequence of an increase in vasomotor tone. This was accompanied by potentiation of the response to PHE as demonstrated by a left shift in the dose-response curve and a decrease in the EC50. Dexamethasone did not potentiate ET-1, but the interplay with the lower baseline temperature resulted in a significantly lower skin temperature for this vasoconstrictor after DEX. The different ET-1 blockers had no effect on ET-1 modulated skin temperatures. CONCLUSIONS: Dexamethasone decreases skin perfusion. This is accompanied by a potentiated alpha1-adrenoceptor agonist response and a greater response to ET-1. POTENTIAL RELEVANCE: Glucocorticoid therapy probably decreases perfusion of the equine hoof. During disease states that already are characterised by hypoperfusion and/or increased levels of circulating catecholamines, glucocorticoid therapy could, according to the vascular model of laminitis, tilt the balance in favour of laminitis.

Drinking salt water enhances rehydration in horses dehydrated by frusemide administration and endurance exercise.

Because the primary stimulus for thirst is an increase in plasma tonicity, we hypothesised that dehydrated horses would drink a greater total volume of fluid voluntarily during the first hour of recovery when they were initially offered salt water. To test this hypothesis, bodyweight (bwt), fluid intake (FI) and [Na+] were measured in 6 Arabian horses offered 3 rehydration solutions. After dehydration was induced by frusemide administration (1 mg/kg bwt, i.v.) followed by 45 km treadmill exercise, water (W), 0.45% NaCl and 0.9% NaCl were offered, in a randomised order, during the initial 5 min after completing exercise. Horses were subsequently placed in a stall and further intake of plain water during the first hour of recovery was measured. By the end of exercise, horses lost 5.2 +/- 0.2, 5.6 +/- 0.3 and 5.7 +/- 0.2% (P>0.05) bwt and FI during the first 5 min of recovery was 10.5 +/- 0.7, 11.6 +/- 0.8 and 11.6 +/- 1.5 l (P>0.05) for W, 0.45% NaCl and 0.9% NaCl, respectively. After 20 min of recovery, [Na+] had decreased with W but remained unchanged from the end exercise values for both saline solutions. During the initial hour of recovery, further water intake was 0.9 +/- 0.4, 5.0 +/- 0.5 and 6.9 +/- 0.7 l (P<0.05) for W, 0.45% NaCl and 0.9% NaCl, respectively. Therefore, total FI was 11.4 +/- 0.5, 16.6 +/- 0.7 and 18.5 +/- 1.7 l (P<0.05) for W, 0.45% NaCl and 0.9% NaCl, respectively, and persisting bwt loss after 60 min of recovery was greater (P<0.05) for W (3.5%) than for the 2 saline solutions (24% for 0.45% NaCl and 1.9% for 0.9% NaCl). In conclusion, providing salt water as the initial rehydration fluid maintained an elevated [Na+] and resulted in greater total FI and recovery of bwt loss during the first hour of recovery, in comparison to offering only plain water.

Potassium and lactate uptake by noncontracting tissue during strenuous exercise.

Efflux of potassium (K+) and lactate (lac-) from active skeletal muscle during high intensity exercise leads to increased plasma [K+] and [lac-] in venous and arterial blood. The exercise-induced increases in these ions in human athletes is ameliorated, in part, by uptake by noncontracting tissue, reflected by an increase in the arteriovenous difference across these tissues from rest to exercise. To test the hypothesis that these ions are taken up by noncontracting tissue in strenuously exercising horses, plasma [K+] and [lac-] were measured in blood samples collected simultaneously from the carotid artery (CA), femoral vein (FV), and transverse facial vein (TFV) in 5 unconditioned Standardbred horses performing 4.5 min of strenuous treadmill exercise. Further, the arteriovenous differences ((a-v)diff) in [K+] and [lac-] were calculated across active tissue (CA-FV) and noncontracting tissue (CA-TFV). After 3 min of strenuous exercise, plasma [K+] in FV blood (mean +/- s.e. 7.1 +/- 0.2 mmol/l) was greater (P<0.05) than in CA blood (6.8 +/- 0.2 mmol/l) and both of these concentrations were greater (P<0.05) than a value of 5.7 +/- 0.1 mmol/l in TFV blood. Similarly, plasma [lac-] increased to 20.6 +/- 0.8 and 21.1 +/- 0.8 mmol/l in CA and FV blood, respectively, and these concentrations were greater (P<0.05) than a value of 19.1 +/- 0.8 mmol/l in TFV blood. The magnitude of the (a-v)diff for both [K+] and [lac-] increased (P<0.05) across active and noncontracting tissue during high-speed exercise but the directions were opposite, supporting release by active tissue and uptake by noncontracting tissue. These results confirm that uptake of K+ and lac- by noncontracting tissue contributes to regulation of the plasma concentrations of these ions in strenuously exercising horses.

Endoscopic assessment of airway function as a predictor of racing performance in Thoroughbred yearlings: 427 cases (1997-2000).

OBJECTIVE: To compare endoscopic findings of the upper portion of the respiratory tract in Thoroughbred yearlings with their subsequent race records to determine whether subjective assessment of airway function may be used as a predictor of future racing performance. DESIGN: Retrospective study. ANIMALS: 427 Thoroughbred yearlings. PROCEDURES: Endoscopic examination findings were obtained from the medical records and the videoendoscopic repository of the Keeneland 1996 September yearling sales. Racing records were requested for the yearlings through the end of their 4-year-old racing season (1997-2000). Twenty-nine measures of racing performance were correlated with endoscopic findings. Subjective arytenoid cartilage movement grades were determined, using a 4-point grading scale (grade 1 = symmetrical synchronous abduction of the arytenoid cartilages; grade 4 = no substantial movement of the left arytenoid cartilage). RESULTS: Of the 427 Thoroughbred yearlings included in this study, 364 established race records, and 63 did not. Opinions regarding suitability for purchase, meeting conditions of the sale, and the presence of epiglottic abnormalities had no significant association with racing performance. Arytenoid cartilage movement grades were significantly associated with many of the dependent variables. However, palatine abnormalities were not predictive of inferior racing performance. CONCLUSIONS AND CLINICAL RELEVANCE: Thoroughbred yearlings with grade-1 and -2 arytenoid cartilage movements had significantly better racing performance as adults, compared with yearlings with grade-3 arytenoid cartilage movements. In contrast, epiglottic and palatine abnormalities were not predictive of inferior racing performance. Therefore, evaluation of laryngeal function, but not epiglottic or palatine abnormalities, using the 4-point grading system, should be the major factor in developing recommendations for prospective buyers.

Identification of sources of Salmonella organisms in a veterinary teaching hospital and evaluation of the effects of disinfectants on detection of Salmonella organisms on surface materials.

OBJECTIVE: To determine sources of Salmonella organisms in a veterinary teaching hospital, compare bacterial culture with polymerase chain reaction (PCR) testing for detection of Salmonella organisms in environmental samples, and evaluate the effects of various disinfectants on detection of Salmonella organisms on surface materials. DESIGN: Prospective study. SAMPLE POPULATION: Fecal samples from 638 hospitalized horses and 783 environmental samples. PROCEDURE: Standard bacterial culture techniques were used; the PCR test amplified a segment of the Salmonella DNA. Five disinfectants were mixed with Salmonella suspensions, and bacterial culture was performed. Swab samples were collected from 7 surface materials after inoculation of the surfaces with Salmonella Typhimurium, with or without addition of a disinfectant, and submitted for bacterial culture and PCR testing. RESULTS: Salmonella organisms were detected in fecal samples from 35 (5.5%) horses. For environmental samples, the proportion of positive bacterial culture results (1/783) was significantly less than the proportion of positive PCR test results (110/783), probably because of detection of nonviable DNA by the PCR test. Detection of Salmonella organisms varied with the surface material tested, the method of detection (bacterial culture vs PCR testing), and the presence and type of disinfectant. CONCLUSIONS AND CLINICAL RELEVANCE: Results of the present study suggested that Salmonella organisms can be isolated from feces of hospitalized horses and a variety of environmental surfaces in a large animal hospital. Although recovery of Salmonella organisms was affected by surface material and disinfectant, bleach was the most effective disinfectant on the largest number of surfaces tested.

An outbreak of salmonellosis among horses at a veterinary teaching hospital.

Between May 1996 and February 1997, 27 horses and a veterinary student at a veterinary teaching hospital developed apparent nosocomial Salmonella Typhimurium infection. The source of the multiple-drug resistant Salmonella Typhimurium was a neonatal foal admitted for treatment of septicemia. A high infection rate (approx 13% of hospitalized horses) coupled with a high case fatality rate (44%) for the initial 18 horses affected led to a decision to close the hospital for extensive cleaning and disinfection. Despite this effort and modification of hospital policies for infection control, 9 additional horses developed nosocomial Salmonella Typhimurium infection during the 6 months after the hospital reopened. Polymerase chain reaction testing of environmental samples was useful in identifying a potential reservoir of the organism in drains in the isolation facility. Coupled with clinical data, comparison of antimicrobial resistance patterns of Salmonella Typhimurium isolates provided a rapid initial means to support or refute nosocomial infection. Although minor changes in the genome of these isolates developed over the course of the outbreak, pulsed-field gel electrophoresis testing further supported that salmonellosis was nosocomial in all 27 horses.

Glycerol hyperhydration in resting horses.

SUMMARY: To determine whether administration of glycerol-containing solutions induces a state of transient hyperhydration in resting euhydrated horses, changes in plasma and urine constituents were measured in four horses for 1 h before and 5 h after nasogastric administration of each of four treatments (Experiment 1). Treatments were applied in a randomized fashion and included: (1) 1.0 g.kg(-)(1)glycerol in 8 L of water (G); (2) 8 L of water (W); (3) 8 L of 0.9% NaCl solution (S); and (4) 1.0 g.kg(-)(1)glycerol in 8 L of 0.9% NaCl solution (GS). In a subsequent study, voluntary water intake was measured hourly for 5 h after nasogastric administration of each treatment (Experiment 2). All treatments produced mild plasma volume expansion ranging from 3.2 to 5.8% in Experiment 1. Administration of glycerol containing solutions increased serum glycerol concentration approximately 100-fold and plasma osmolality (P(osm)) by approximately 10 mOsm/kg and resulted in a tendency towards increased renal water conservation despite increased osmole excretion. In contrast, W treatment decreased plasma and urine osmolality and was accompanied by increased urine production and decreased renal water conservation. Plasma and urine osmolality, as well as renal osmole and water excretion, were unchanged after S administration. In Experiment 2, horses treated with GS voluntarily drank an additional 5.2 +/- 0.9 L of water during the initial hour following nasogastric administration of 8 L of solution. Voluntary water intake with the other treatments was less than 1.0 L for the entire 5 h observation period. Collectively, the results of both experiments suggest that administration of glycerol in saline would produce transient hyperhydration in resting euhydrated horses by enhancing renal water conservation and stimulating voluntary water intake.

Effects of electrolyte and glycerol supplementation on recovery from endurance exercise.

Incomplete recovery from endurance exercise after an overnight rest period is reflected by persisting weight loss and an elevated plasma aldosterone concentration, even in successful competitors. To determine whether supplementation with high doses of electrolytes, with or without glycerol, enhances recovery, the following were measured in 6 Arabian horses before and after completion of a 60 km treadmill exercise test simulating an endurance ride and after 12, 24, 48, and 72 h of recovery: bodyweight; plasma osmolality; plasma concentrations of protein, electrolytes, aldosterone and cortisol; and urine and faecal electrolyte concentrations. Before and during the exercise test, horses were supplemented with a total of 2.4 ml/kg bwt of water (W); 0.2 g/kg bwt KCl and 0.4 g/kg bwt NaCl in 2.4 ml/kg bwt of water (E); or 0.2 g/kg bwt KCl and 0.4 g/kg bwt NaCl in 2.4 ml/kg bwt (3 g/kg bwt) of glycerol (GE). Although weight loss after completion of the simulated ride was greater (P < 0.01) for W (3.2%) than for E and GE (1.0 and 0.9%, respectively), horses supplemented with E or GE experienced further weight loss by 24 h after the simulated ride (2.2 and 2.1% for E and GE, respectively) while bodyweight with W remained unchanged (3.0%) from the finish value. After 48 h of recovery, bodyweight was not different from the starting values with E and GE but remained decreased (P < 0.01) with W throughout the recovery period (2.2% persisting weight loss after 72 h of recovery). Plasma osmolality and plasma Na+ and Cl- concentrations increased (P < 0.01) and plasma protein concentration decreased (P < 0.01) after the exercise test with E and GE but were unchanged with W. Plasma osmolality and protein and electrolyte concentrations returned to pre-exercise values within 12 h of recovery with the exception of a persistent increase in plasma Na+ concentration with GE. The greatest plasma aldosterone concentration was measured after 12 h of recovery with W (1357 pmol/l) and was greater (P < 0.02) than that with E and GE (24 and 304 pmol/l, respectively). Urine production during the simulated ride increased (P < 0.01) with GE and resulted in loss of approximately 20% and essentially 100% of supplemented Na+ and K+, respectively. In contrast, electrolyte losses in faeces were unaffected by electrolyte or glycerol supplementation. In conclusion, supplementation with high doses of electrolytes as hypertonic oral pastes attenuated weight loss during a simulated endurance ride (by enhancing voluntary water intake); however, it did not prevent development of significant weight loss during the initial 24 h of recovery. Glycerol administration resulted in no benefits, and actually increased urine electrolyte losses, in comparison to supplementation with electrolytes alone.

Electrolyte and glycerol supplementation improve water intake by horses performing a simulated 60 km endurance ride.

To replace electrolytes lost in sweat during endurance competitions, riders frequently supplement horses with hypertonic oral electrolyte pastes. To determine whether this practice and concurrent administration of the so-called hyperhydrating substance glycerol are of benefit, weight loss, voluntary water intake, plasma osmolality, and plasma protein and electrolyte concentrations were measured in 6 Arabian horses supplemented with a total of 2.4 ml/kg bwt of water (W); 0.2 g/kg bwt KCl and 0.4 g/kg bwt NaCl in 2.4 ml/kg bwt of water (E); or 0.2 g/kg bwt KCl and 0.4 g/kg bwt NaCl in 2.4 ml/kg bwt (3 g/kg bwt) of glycerol (GE) before and during a treadmill exercise test simulating a 60 km endurance ride. Weight loss was greater (P < 0.01) with W (3.2%) than with E and GE (1.0% and 0.9%, respectively) and was associated (r = -0.85, P < 0.0001) with less (P < 0.01) water intake with W (12.2 l) than with E or GE (23.5 l and 25.8 l, respectively). Plasma osmolality increased to a greater extent (P < 0.01) with GE than with E and was unchanged with W. In contrast, plasma protein concentration decreased (P < 0.01) in the later stages of the simulated ride with E and GE, reflecting plasma volume expansion, but remained unchanged with W. Plasma Na+ and Cl- concentrations increased (P < 0.01) with E and GE and were greater (P < 0.01) than values for W during the second half of the 60 km simulated ride. Despite administration are large amounts of potassium with E and GE, plasma K+ concentration was decreased (P < 0.01) at the end of the rest breaks during the simulated ride and after 60 min of recovery with all supplements. Adverse effects of administration of hypertonic oral electrolyte pastes were not observed and decreases in plasma protein concentration within 30 min after drinking with all supplements suggested that intestinal absorption was well maintained during the simulated endurance ride. In conclusion, electrolyte supplementation in the form of hypertonic oral pastes improved water intake during endurance exercise but concurrent glycerol administration provided no additional benefits in comparison to supplementation with electrolytes alone.

Efficacy of the intranasal route for administration of anesthetic agents to adult rabbits.

Anesthetic agents were administered to adult rabbits by using the intranasal route. Six sedative or anesthetic protocols were studied as follows: group 1 (n = 12), 2.0 mg midazolam/kg body weight (BW); group 2 (n = 8), 25.0 mg ketamine/kg BW; group 3 (n = 8), 10 mg of combination of tiletamine and zolazepam/kg BW; group 4 (n = 10), 3 mg xylazine/kg BW and 10 mg ketamine/kg BW; group 5 (n = 8), 1.0 mg midazolam/kg BW and 25 mg ketamine/kg BW; and group 6 (n = 6), 0.3 ml of a combination of fentanyl and droperidol/kg BW. All drugs were diluted to a final volume of 0.4 ml/kg BW and an equal volume was administered with a catheter-tipped syringe into each nostril. Time to onset and duration of sedation or anesthesia were recorded. Muscle relaxation was graded as poor, fair, or excellent on the basis of flexibility of limbs. Presence or absence of a toe pinch response was recorded. Heart rate, respiratory rate, and hemoglobin saturation were measured before and at 5-min intervals after drug administration. The mean onset times for groups 1, 2, 3, 4, and 5 were 3.0, 1.2, 2.5, 2.0, and 0.8 min, respectively. The mean duration of action was 24.6, 36.7, 44.4, 35.2, and 52.5 min for midazolam, ketamine, tiletamine/zolazepam, xylazine/ketamine, and midazolam/ketamine, respectively. All protocols resulted in a significant decrease in respiratory rate. Hemoglobin saturation decreased in all groups except group 1.(ABSTRACT TRUNCATED AT 250 WORDS)

Practical considerations for maintaining germplasm in maize.

The main goals of genetic resource management are to acquire, maintain, distribute, characterize, regenerate, preserve, evaluate, and utilize the genetic diversity of crops and their wild relatives. The objectives of this study for ex-situ conservation of maize (Zea mays L.) are to review and describe: (1) practical regeneration methods that are based on population genetic theory; (2) practical problems encountered in choosing core subsets of a maize collection. Whenever possible, regeneration procedures should control the number of pollen parents (male gametes; through controlled hand pollination) and the number of female parent gametes (by harvesting equal numbers of kernels from each seed plant). When the number of pollen and seed parents are controlled during regeneration, the effective population size (Ne) is twice the size of the original population (N). Examples of practical methods for controlling the number of male and female parents are presented. The procedure involves random-paired plant crosses and taking equal numbers of seeds from each maize ear. To form a core subset, accessions of a maize race are subdivided through a stratified sampling procedure. Delineation of a core subset from a Tuxpeño racial collection is described as an example.

In vitro effects of alpha 2-adrenergic receptor stimulation on cholinergic contractions of equine distal airways.

In horses with noninduced, reversible airway obstruction (heaves), pulmonary function is improved after sedation with the alpha 2-adrenergic agonist xylazine. The mechanism of this effect is undetermined. Because the predominant excitatory innervation of equine airways is cholinergic, the influence of alpha 2-adrenergic receptor stimulation on the response of isolated distal airways to cholinergic stimulation was determined. Distal bronchial segments from 22 healthy horses were suspended in isolated organ baths where their mechanical responses to various stimuli could be studied. Each tissue was incubated with one of several concentrations of clonidine, clonidine vehicle, or clonidine plus tolazoline. Then, the contractile response of the tissues to either cumulative acetylcholine (ACh) addition or cumulative electrical field stimulation (EFS) was recorded. All contractile responses evoked by EFS were mediated through stimulation of cholinergic airway nerves. Clonidine had no effect on the contractile response of distal airway segments to exogenous ACh. However, clonidine (at concentrations > 10(-5) M) significantly (P < 0.05) diminished the contractile response of the distal airway segments to EFS. This inhibitory effect of clonidine was not observed in the presence of tolazoline. Similar results were observed when the less-selective alpha 2-adrenergic agonist xylazine was exposed to the isolated segments instead of clonidine. Because EFS-but not exogenous ACh-induced contractions were inhibited, alpha 2-adrenergic receptor stimulation apparently causes presynaptic inhibition of the cholinergic nerves innervating distal portions of the bronchi of horses.

Cardiovascular responses to exogenous platelet-activating factor (PAF) in anesthetized ponies, and the effects of a PAF antagonist, WEB 2086.

The effects of exogenous platelet-activating factor (PAF) were determined in anesthetized ponies. Administration of PAF induced a decrease in cardiac index that resulted in systemic hypotension. This was followed by tachycardia, hypertension, and a return of cardiac index to baseline. Pulmonary arterial pressure increased markedly because of pulmonary vasoconstriction. Exogenous PAF also caused leukopenia and thrombocytopenia. The specific PAF receptor antagonist (WEB 2086) blocked all PAF-induced changes. Flunixin meglumine, a cyclooxygenase inhibitor, abolished the pulmonary hypertension and tachycardia, and attenuated the systemic hypotension but did not change the PAF-induced peripheral cellular changes. The PAF antagonist also inhibited platelet aggregation induced by PAF in vitro. The PAF-induced changes are similar to those reported after endotoxin exposure in horses.

Statistical genetic considerations for maintaining germ plasm collections.

One objective of the regeneration of genetic populations is to maintain at least one copy of each allele present in the original population. Genetic diversity within populations depends on the number and frequency of alleles across all loci. The objectives of this study on outbreeding crops are: (1) to use probability models to determine optimal sample sizes for the regeneration for a number of alleles at independent loci; and (2) to examine theoretical considerations in choosing core subsets of a collection. If we assume that k-1 alleles occur at an identical low frequency of p0 and that the k(th) allele occurs at a frequency of 1-[(k-1)p0], for loci with two, three, or four alleles, each with a p0 of 0.05, 89-110 additional individuals are required if at least one allele at each of 10 loci is to be retained with a 90% probability; if 100 loci are involved, 134-155 individuals are required. For two, three, or four alleles, when p0 is 0.03 at each of 10 loci, the sample size required to include at least one of the alleles from each class in each locus is 150-186 individuals; if 100 loci are involved, 75 additional individuals are required. Sample sizes of 160-210 plants are required to capture alleles at frequencies of 0.05 or higher in each of 150 loci, with a 90-95% probability. For rare alleles widespread throughout the collection, most alleles with frequencies of 0.03 and 0.05 per locus will be included in a core subset of 25-100 accessions.