Organic dust exposure increases mast cell tryptase in bronchoalveolar lavage fluid and airway epithelium of heaves horses.

BACKGROUND: Mast cell degranulation is believed to act as a key event in initiating and maintaining airway response to allergen challenge in human asthma. It is hypothesized that the mast cell may play a similar role in equine heaves, which shares many similarities with occupational dust-induced asthma. OBJECTIVE: The aim of this study was to quantify the mast cell proteinase tryptase in bronchoalveolar lavage fluid (BALF) from control and heaves-susceptible horses and to investigate tryptase mRNA and protein expression in pulmonary mast cells. METHODS: Equine BALF tryptase concentrations were determined by ELISA from control and heaves-susceptible horses pre and post 24 h hay/straw challenge (HSC). Tryptase mRNA and protein expression were investigated by quantitative PCR and immunohistochemistry in bronchial and bronchiolar tissue samples of control and heaves-susceptible horses. RESULTS: Both control and heaves-susceptible horses had significantly increased BALF tryptase concentrations following HSC (P=0.003 and 0.034, respectively). Increased numbers of tryptase-expressing intra-epithelial mast cells were demonstrated in heaves horses, but not controls, following challenge (P=0.02). Bronchiolar tissue from heaves horses removed from challenge contained significantly lower tryptase transcripts than that from control horses (P=0.02). CONCLUSION: Mast cell degranulation and tryptase release into the airways occur following HSC of control and heaves-susceptible horses. The greater number of mast cells available in the bronchiolar epithelium of heaves horses may be clinically significant in the pulmonary inflammatory response of heaves.

Early onset airway obstruction in response to organic dust in the horse.

Equine recurrent airway obstruction (RAO) has been used as a naturally occurring model of human asthma. However, it is unknown whether there is an early-phase response in RAO. The aim of this study was to determine whether exposure to organic dust induces immediate changes in lung function in RAO-affected horses, which could be mediated by airway mast cells. Six RAO-affected horses in remission and six control horses were challenged with hay-straw dust suspension by nebulization. Total respiratory resistance at 1 Hz, measured by forced oscillation, was increased from 0.62 +/- 0.09 cmH(2)O.l(-1).s (mean +/- SE) to 1.23 +/- 0.20 cmH(2)O.l(-1).s 15 min after nebulization in control horses (P = 0.023) but did not change significantly in the RAO group. Total respiratory reactance at 1 Hz (P = 0.005) was significantly lower in the control horses (-0.77 +/- 0.07 cmH(2)O.l(-1).s) than in the RAO group (-0.49 +/- 0.04 cmH(2)O.l(-1).s) 15 min after nebulization. Bronchoalveolar lavage fluid (BALF) histamine concentration was significantly elevated 10 and 20 min postnebulization in control horses but not in RAO horses. Minimum reactance at 1 Hz in the early postnebulization period significantly correlated with both prechallenge BALF mast cell numbers (r = -0.65, P = 0.02) and peak BALF histamine concentration postnebulization (r = -0.61, P = 0.04). In conclusion, RAO horses, unlike human asthmatic patients, do not exhibit an early-phase response. However, healthy control horses do demonstrate a mild but significant early (<20 min) phase response to inhaled organic dust. This response may serve to decrease the subsequent dose of dust inhaled and as such provide a protective mechanism, which may be compromised in RAO horses.

cDNA cloning and substrate specificity of equine tryptase, a possible mediator in equine heaves.

BACKGROUND: Mast cell mediators are believed to play a central role in inflammatory lung disorders such as human allergic and occupational asthma. Equine heaves is characterized by reversible neutrophilic airway inflammation and airway obstruction, primarily due to bronchospasm and mucus hypersecretion, following exposure of susceptible horses to organic stable dusts. As such, heaves shares many similarities with human occupational dust-induced asthma and therefore it is proposed that mast cells may also be implicated in the pathogenesis of heaves. Tryptase, a mast cell-specific proteinase, can be used as an indicator of biological mast cell activity. OBJECTIVE: The aim of this study was to determine the cDNA sequence of equine tryptase and to investigate its substrate specificity in order to rationalize its enzymatic activity. METHODS: RT-PCR cloning was used to sequence equine tryptase. Substrate specificity of equine tryptase was investigated using arginine and lysine containing substrates. RESULTS: The cDNA and deduced amino acid (Aa) sequences for equine tryptase shared strong identity with other tryptases. Unusually for a trypsin-like proteinase however, equine tryptase has alanine at residue 216, rather than glycine, which confers increased arginine substrate specificity in vitro and may restrict fibrinogenolysis in vivo. CONCLUSION: Cloning and sequencing of the mast cell proteinase equine tryptase will allow molecular probing of its expression in the lung of control and heaves-affected horses. Further work is warranted to determine the biological relevance of the unique alanine 216 substitution in the molecular sequence of the equine tryptase substrate-binding pocket.

Changes in selected physiological and laboratory measurements in elite horses competing in a 160 km endurance ride.

REASON FOR PERFORMING STUDY: Limited information exists about the physiological changes and clinical problems that occur in elite horses competing in high-speed 160 km endurance races. OBJECTIVES: To provide initial data describing changes in physiological and laboratory measurements in horses competing in a high-speed, 160 km endurance race under temperate conditions and to compare data between horses that successfully completed the race and those that failed to finish. METHODS: Body mass (BM) was measured, blood samples were collected, and veterinary examinations performed on horses before, during, and at the finish of a CEI*** 160 km endurance race. RESULTS: Of 36 horses participating in the study, 22 (61%) completed the race. Twelve horses were eliminated for lameness and 2 for persistent heart rate elevation. Mean speed of finishers was 15.2 km/h. Mean +/- s.d. BM loss of finishers at the end of the race (5.7 +/- 2.6%) was not different (P = 0.58) from BM loss of nonfinishers at elimination (6.7 +/- 34%). Similarly, there were no significant differences in heart rate or veterinary assessment of hydration at the race end for finishers as compared to the elimination point for nonfinishers. PCV increased while sodium, chloride and potassium concentrations decreased with exercise but differences between finishers and nonfinishers were not detected. In contrast, both total and ionised calcium concentrations decreased in successful horses but remained unchanged in nonfinishers. CONCLUSIONS: Elite endurance horses are more likely to be eliminated from competition for lameness than metabolic problems; however, it remains unclear whether these conditions are entirely distinct. The magnitude of the decrease in sodium concentration in both finishers and nonfinishers was greater than in previous reports of 160 km rides. POTENTIAL RELEVANCE: These data should be of use for both organisers and participants in elite 160 km endurance races. The tendency toward hyponatraemia as well as the difference in calcium concentrations between finishers and nonfinishers warrant further study.

Standing oral extraction of cheek teeth in 100 horses (1998--2003).

REASONS FOR PERFORMING STUDY: Extraction of cheek teeth (CT) by the conventional repulsion technique requires general anaesthesia and carries a high rate of post operative complications. Consequently, an alternative method of extraction, i.e. orally in standing horses, was evaluated. HYPOTHESIS: The need for and risks of general anaesthesia could be avoided and post extraction sequelae reduced by performing extractions orally in standing horses. METHODS: One hundred mainly younger horses (median age 8, range 2-18 years) with firmly attached CT that required extraction because of apical infections, displacements, diastemata, idiopathic fractures and the presence of supernumerary CT had the affected teeth (n = 111) extracted orally under standing sedation. Follow-up information was obtained for all cases, a median of 16 months later. RESULTS: Oral extraction was successful in 89 horses and unsuccessful in 11 due to damage to the CT clinical crown (n = 9) during extraction, for behavioural reasons (n = 1) and because the apex of a partly extracted CT fell back into the alveolus following sectioning (n = 1). Predispositions to extraction-related CT fractures were present in 5 of the 9 cases, i.e. advanced dental caries (n = 2) and pre-existing 'idiopathic' fractures (n = 3). The iatrogenically fractured CT were later repulsed under standing sedation (n = 3) and under general anaesthesia (n = 6). Eighty-one of the remaining 89 horses had successful oral CT extraction with no or minimal intra- or post operative complications occurring. Post operative complications in the other 8 cases included post extraction alveolar sequestration (n = 3), alveolar sequestration and localised osteomyelitis (n = 1), localised osteomyelitis (n = 1), incorporation of alveolar packing material into alveolar granulation tissue (n = 1), and nasal discharge due to continued intranasal presence of purulent food material (n = 1) and to ongoing sinusitis (n = 1). The above sequelae were treated successfully in all cases, with general anaesthesia required in just one case. Following oral extraction, significantly (P<0.001) fewer post operative problems developed in 54 horses with apically infected CT in comparison with 71 previous cases that had repulsion of apically infected CT at our clinic. CONCLUSIONS AND POTENTIAL RELEVANCE: Oral extraction of cheek teeth is a successful technique in the majority of younger horses with firmly attached CT and greatly reduces the post operative sequelae, compared with CT repulsion. Additionally, the costs and risks of general anaesthesia are avoided. Further experience and refinement in the described protocol could potentially increase the success of this procedure and also reduce the incidence of post operative sequelae.

Long-term survey of laryngoplasty and ventriculocordectomy in an older, mixed-breed population of 200 horses. Part 1: Maintenance of surgical arytenoid abduction and complications of surgery.

REASONS FOR PERFORMING STUDY: Laryngoplasty (LP) is currently the most common surgical treatment for equine laryngeal paralysis, however, there have been no reports quantifying the degree of retention of arytenoid abduction following LP. Additionally, the complications of LP have been poorly documented. OBJECTIVES: To record the degree of arytenoid abduction retention following LP and to accurately document all complications of surgery. METHODS: A study (1986-1998) of 200 horses of mixed breed and workload, median age 6 years (prospective 136 cases and retrospective 64 cases) undergoing LP (using 2 stainless steel wires) and combined ventriculocordectomy was undertaken; 198 owners completed questionnaires, a median of 19 months following surgery. The degree of arytenoid abduction achieved was endoscopically, semi-quantitatively evaluated using a 5-grade system, at 1 day, 7 days, and 6 weeks after surgery. RESULTS: On the day following LP, 62% of horses had good (median grade 2) arytenoid abduction, 10% had excessive (grade 1), and 5% had minimal (grade 4) abduction (overall-median grade 2). Due to progressive loss of abduction, moderate (median grade 3, range 1-5) abduction was present overall at 1 and 6 weeks after LP. Further surgery was required to re-tighten prostheses in 10% of cases with excessive loss of abduction, or to loosen prostheses in 7% of horses which had continuing high levels of LP abduction and significant post operative dysphagia. LP wound problems (mainly seromas and suture abscesses) were reported to last < 2 weeks in 9% of cases, < 4 weeks in 4% and > 4 weeks in 4%. The (partially sutured) laryngotomy wounds discharged post operatively for < 2 weeks in 22% of cases, < 4 weeks in 7% and for > 4 weeks in 2%. Coughing occurred at some stage post operatively in 43% of cases and its presence correlated significantly with the degree of surgical arytenoid abduction. This coughing occurred during eating in 24% of cases and was not associated with eating (or dysphagia) in the other 19% of cases. Chronic (> 6 months duration) coughing occurred in 14% of cases, but appeared to be due to intercurrent pulmonary disease in half of these horses. CONCLUSIONS: Suturing the cricotracheal membrane allows most laryngotomy wounds to heal quickly. Laryngoplasty wound problems were of little long-term consequence when stainless steel wire prostheses were used. POTENTIAL RELEVANCE: A significant loss of LP abduction occurs in most horses in the 6 weeks following surgery and efforts should be made to find ways to prevent such loss. However, excessive LP abduction is associated with post operative dysphagia and coughing.

Long-term survey of laryngoplasty and ventriculocordectomy in an older, mixed-breed population of 200 horses. Part 2: Owners' assessment of the value of surgery.

REASONS FOR PERFORMING STUDY: There is continuing debate on the clinical benefit of laryngoplasty (LP) in the treatment of equine laryngeal paralysis. In particular, there is little information available on the clinical value of this surgery in older horses, in non-Thoroughbreds or in sports or pleasure horses; nor on the relationship between the degree of LP abduction achieved and the clinical value of LP. OBJECTIVES: To evaluate the owner's assessment of the value of LP (and combined ventriculocordectomy) in an older, mixed breed and mixed workload population of horses and to also assess the relationship between the degree of LP abduction present and the clinical value of this surgery. METHODS: A study (1986-1998) of 200 horses of mixed breed and workload, median age 6 years (prospective 136 cases and retrospective 64 cases) undergoing LP and ventriculocordectomy was undertaken. The degree of laryngoplasty abduction achieved and maintained was semiquantitatively evaluated using a 5-grade system. RESULTS: A survey of 198 owners, a median of 19 months after LP surgery, showed that 91% of cases had returned to full work and 3% to reduced work, including 95% of horses with good (Grade 2) laryngoplasty abduction at 6 weeks after surgery, 91% with moderate (Grade 3) abduction, 88% of cases with minsimal (Grade 4) abduction and just 25% of cases with total loss of surgical abduction (Grade 5). Once back in work, 73% of cases were reported to make no abnormal exercise-related noises ('noises') at exercise, with reduced 'noises' reported in some of the 21% of horses that still made 'noises'. In the other 6% of cases, owners were unsure if 'noises' were present. The absence or presence of 'noises' once back at work correlated significantly with the degree of surgical arytenoid abduction present at 6 weeks after LP. Once back in work, 'noises' were less commonly detected in sports and pleasure horses (absent in 76% of cases) than in racehorses (absent in 60%) of cases. Exercise performance following surgery was reported to be markedly increased in 75% of cases, with 10% showing no difference in exercise performance, 3% showing worse performance, and owners unsure of any effect on exercise performance in 13% of cases. Improved work was reported in 70% of sports and pleasure horses and in 67% of racehorses. Overall, 86% of owners believed LP was worthwhile, 7% believed it was not worthwhile and 6% were unsure of its value. Surgery was reported to be of most benefit to sports horses (e.g. reported worthwhile for 100% of show jumpers) and of least benefit to National Hunt (long distance) racehorses where 71% of owners believed it to be worthwhile. CONCLUSIONS AND POTENTIAL RELEVANCE: Most horse-owners believe that LP and combined ventriculocordectomy are of clinical value, especially in sports and pleasure horses. Large scale physiological studies on clinical cases, pre- and post operatively, are required to examine more critically the value of such surgery.