Mild-moderate equine asthma: A scoping review of evidence supporting the consensus definition.

Current consensus defines mild-moderate equine asthma (mEA; previously inflammatory airway disease) by a hierarchy of indicators of lung pathology: cough, poor performance, increased tracheobronchial mucus, inflammatory bronchoalveolar lavage (BAL) cytology and pulmonary dysfunction. Exclusion criteria include fever, systemic disease, or increased resting respiratory effort. The aim of this review was to inform future research by identifying gaps, strengths and weaknesses in the current body of evidence supporting this consensus-proposed definition. Objectives were to critique evidence supporting the inclusion of each diagnostic indicator in the case definition, by summarising and evaluating evidence for its association with higher-level indicators of lung inflammation. Searches of three databases identified 2275 articles relating to mEA or its diagnostic indicators, from which 298 full-text articles were screened and 45 reviewed in full. Studies (n = 44) had been performed worldwide in clinics, hospitals, racetracks, yards or research herds, in 6092 horses. Studies were predominantly opportunistic observational (n = 13/44: 29.5%) or cross-sectional (n = 11/44; 25%). The median number of horses per study was 74. Where breed and use were reported most were Thoroughbreds (58.2%; 2730/4688) and racehorses (72.8%; n = 3960/5439). Domains rated as high risk of bias in almost 50% of articles were 'study power' and 'masking'. Heterogeneity in clinical and laboratory measures precluded meta-analysis. Evidence was more consistent for certain pairwise relationships (e.g., between cough and tracheobronchial mucus) than others (e.g., BAL cytology and lung function). Findings highlight the need for increased standardisation of diagnostic methods and reporting to facilitate future systematic review and meta-analysis.

Dust exposure and pulmonary inflammation in Standardbred racehorses fed dry hay or haylage: A pilot study.

Respirable dust exposure is linked to airway inflammation in racehorses. Feeding haylage may reduce dust exposure by 60-70%. The objective of this study was to compare dust exposure, airway cytology, and inflammatory cytokine concentrations between horses fed haylage or hay over 6 weeks while in training. Seven healthy Standardbred horses were randomly assigned to be fed alfalfa hay (n = 3) or grass-alfalfa mix haylage (n = 4) for six weeks while training on a treadmill. Dust exposure was measured gravimetrically at the breathing zone. Endotoxin and ß-glucan concentrations in respirable dust were measured. Bronchoalveolar lavage fluid (BALF) cytology was determined at baseline and after 2, 4, and 6 weeks. Cytokine concentrations (interferon-γ, tumor necrosis factor-α, and interleukin-4) were measured in BALF at baseline and week 6. The effect of forage on exposure, airway cytology and cytokines were evaluated using generalized linear mixed models. Respirable dust and ß-glucan exposures were lower in horses fed haylage than hay (0.02 ± 0.001 mg/m3 vs. 0.06 ± 0.01 mg/m3; P = 0.03, and 69 ± 18 pg/m3 vs. 160 ± 21 pg/m3; P = 0.02, respectively). In horses eating haylage, BALF neutrophil proportion decreased between baseline (2.2 ± 0.5%), week 2 (0.8 ± 0.3%; P = 0.01) and week 6 (0.7 ± 0.2%; P = 0.03). By week 6, horses fed haylage had lower BALF neutrophilia than horses fed hay (4.0 ± 0.7 %; P = 0.0004). Interleukin-4 concentration in BALF was higher at week 6 (14.4 ± 4.6 pg/mL) in horses fed hay compared to baseline (2.9 ± 4.6 pg/mL; P = 0.007). In conclusion, feeding haylage instead of hay to horses in training can reduce exposure to respirable irritants and mitigate airway neutrophilia.

Antigen array for serological diagnosis and novel allergen identification in severe equine asthma.

Severe equine asthma (sEA), which closely resembles human asthma, is a debilitating and performance-limiting allergic respiratory disorder which affects 14% of horses in the Northern Hemisphere and is associated with increased allergen-specific immunoglobulin E (IgE) against a range of environmental proteins. A comprehensive microarray platform was developed to enable the simultaneous detection of allergen-specific equine IgE in serum against a wide range of putative allergenic proteins. The microarray revealed a plethora of novel pollen, bacteria, mould and arthropod proteins significant in the aetiology of sEA. Moreover, the analyses revealed an association between sEA-affected horses and IgE antibodies specific for proteins derived from latex, which has traditionally been ubiquitous to the horse's environment in the form of riding surfaces and race tracks. Further work is required to establish the involvement of latex proteins in sEA as a potential risk factor. This work demonstrates a novel and rapid approach to sEA diagnosis, providing a platform for tailored management and the development of allergen-specific immunotherapy.

Efficacy of Inhaled Levalbuterol Compared to Albuterol in Horses with Recurrent Airway Obstruction.

BACKGROUND: The (R)-enantiomer of racemic albuterol (levalbuterol) has bronchodilatory properties whereas the (S)-enantiomer causes adverse effects in human airways, animal models, and isolated equine bronchi. Levalbuterol is commercially available and improves pulmonary function of asthmatic patients with a longer duration of effect than albuterol. OBJECTIVE: To determine the dose at which inhaled levalbuterol produces maximal bronchodilatory effect (EDmax) and determine its duration of action in recurrent airway obstruction (RAO)-affected horses in comparison to racemic albuterol. ANIMALS: Nine horses with inducible and reversible RAO. METHODS: Randomized, crossover trial. Horses were challenged with moldy hay to induce airway obstruction. Horses were treated with nebulized albuterol or levalbuterol chosen randomly. Pulmonary function testing (PFT) was measured before and for up to 3 hours after bronchodilatation challenge. Maximum change in transpulmonary pressure (DPmax ) was measured to assess the dose effect and duration of action of each drug. After a 24 hours washout period, the bronchodilatation challenge was repeated with the second bronchodilator. RESULTS: The duration of effect was 60 minutes for albuterol and 120 minutes for levalbuterol. The dose of bronchodilator EDmax was not significantly different between albuterol and levalbuterol (EDmax = 125.0 [125-125 µg] and EDmax = 188 [125-188 µg] respectively; P = .068). The magnitude of bronchodilatation was not significantly different between the 2 treatments (61.1 and 59.9% decrease in DPmax for albuterol and levalbuterol respectively; P = .86). CONCLUSIONS AND CLINICAL IMPORTANCE: Levalbuterol is as effective a bronchodilator as albuterol; although levalbuterol lasts twice as long as albuterol, its duration of action is still too short to make it practical for RAO treatment.

Inflammatory Airway Disease of Horses--Revised Consensus Statement.

The purpose of this manuscript is to revise and update the previous consensus statement on inflammatory airway disease (IAD) in horses. Since 2007, a large number of scientific articles have been published on the topic and these new findings have led to a significant evolution of our understanding of IAD.

Secretoglobin and Transferrin Expression in Bronchoalveolar Lavage Fluid of Horses with Chronic Respiratory Disease.

BACKGROUND: Lower expression of secretoglobin and transferrin has been found in the bronchoalveolar lavage fluid (BALF) of a small number of horses with experimentally induced signs of recurrent airway obstruction (RAO) compared to healthy controls. HYPOTHESIS/OBJECTIVES: Secretoglobin and transferrin BALF expression will be similarly decreased in horses with naturally occurring clinical signs of RAO and in horses with experimentally induced clinical signs of RAO as compared to healthy controls and intermediate in horses with inflammatory airway disease (IAD). ANIMALS: Recurrent airway obstruction-affected and control horses were subjected to an experimental hay exposure trial to induce signs of RAO. Client-owned horses with a presumptive diagnosis of RAO and controls from the same stable environments were recruited. METHODS: Pulmonary function and BALF were evaluated from control and RAO-affected research horses during an experimental hay exposure trial (n = 5 in each group) and from client-owned horses (RAO-affected horses, n = 17; IAD-affected horses, n = 19; healthy controls, n = 5). The concentrations of secretoglobin and transferrin in BALF were assessed using Western blots. RESULTS: Naturally occurring and experimentally induced RAO horses had similar decreases in BALF transferrin expression, but secretoglobin expression was most decreased in naturally occurring RAO. Secretoglobin and transferrin expression were both lower in BALF of RAO-affected horses than in IAD-affected and control horses. CONCLUSIONS AND CLINICAL IMPORTANCE: Secretoglobin and transferrin expression is decreased in BALF of RAO-affected horses after both experimental and natural exposure. Secretoglobin and transferrin likely play clinically relevant roles in the pathophysiology of RAO, and may thus be used as biomarkers of the disease.

Exercise induced pulmonary hemorrhage in horses: American College of Veterinary Internal Medicine consensus statement.

BACKGROUND: Published studies of exercise-induced pulmonary hemorrhage (EIPH), when assessed individually, often provide equivocal or conflicting results. Systematic reviews aggregate evidence from individual studies to provide a global assessment of the quality of evidence and to inform recommendations. OBJECTIVES: Evaluate evidence to determine: if EIPH adversely affects the health, welfare or both of horses; if EIPH affects the athletic capacity of horses; the efficacy of prophylactic interventions for EIPH; and if furosemide affects the athletic capacity of horses. ANIMALS: None. MATERIALS AND METHODS: Systematic review. A panel of 7 experts was formed to assess evidence in the peer reviewed literature addressing each of the 4 objectives. Methodology followed that of the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE). Publications were assessed for quality of evidence by working groups of the panel, and a summary of findings was presented in tables. Recommendations were based on quality of evidence and were determined by a vote of the panel. RESULTS: Much of the evidence was of low to very low quality. Experimental studies frequently lacked adequate statistical power. There was moderate to high quality evidence that EIPH is progressive, is associated with lung lesions, that it adversely affects racing performance, that severe EIPH (Grade 4) is associated with a shorter career duration, that furosemide is efficacious in decreasing the incidence and severity of EIPH, and that administration of furosemide is associated with superior race performance. CONCLUSIONS AND CLINICAL SIGNIFICANCE: Strong recommendation that EIPH be considered a disease and a weak recommendation for use of furosemide in management of racehorses with EIPH.

Omega-3 fatty acid supplementation provides an additional benefit to a low-dust diet in the management of horses with chronic lower airway inflammatory disease.

BACKGROUND: Omega-3 polyunsaturated fatty acid (PUFA) may benefit humans and animals with chronic inflammatory diseases. HYPOTHESIS: Omega-3 PUFA supplementation improves clinical signs, lung function, and airway inflammation in horses with recurrent airway obstruction (RAO) and inflammatory airway disease (IAD). ANIMALS: Eight research horses and 35 client-owned horses. METHODS: A pilot study examined the dose of PUFA that can alter plasma PUFA composition. Then, a randomized, controlled clinical trial was performed in horses with RAO and IAD. Horses were fed a complete pelleted diet with no hay and randomly assigned to 1 of 3 daily treatments for 2 months: 30 or 60 g of the supplement or 30 g of placebo. Clinical signs, lung function, plasma PUFA composition, and bronchoalveolar lavage fluid (BALF) cytology were evaluated. Data were expressed as median (25-75th percentiles). P < .05 was considered significant. RESULTS: Polyunsaturated fatty acid supplementation resulted in increased plasma docosahexaenoic acid (DHA) that peaked at 4 weeks. Clinical improvement was noted in all horses involved in the clinical trial, but the group that received PUFA had greater improvement in clinical signs (cough score improved 60%), lung function (respiratory effort decreased 48%), and BALF (neutrophils decreased from 23 to 9%) when compared to placebo (cough score improved 33%, respiratory effort decreased 27%, BALF neutrophils increased from 11 to 17%; P < .05). CONCLUSIONS AND CLINICAL IMPORTANCE: Feeding horses with RAO and IAD a PUFA supplement containing 1.5-3 g DHA for 2 months provides an additional benefit to low-dust diet.

Investigating the link between particulate exposure and airway inflammation in the horse.

Inhalant exposure to airborne irritants commonly encountered in horse stables is implicated in the pathogenesis of inflammatory airway disease (IAD) and recurrent airway obstruction (RAO), non-infectious, inflammatory pulmonary disorders that impact the health and performance of horses across all equine disciplines. IAD and RAO have overlapping clinical, cytological, and functional manifestations of the pulmonary response to organic dust and noxious gases encountered in the barn environment. Study of these diseases has provided important but incomplete understanding of the effect of air quality upon the respiratory health of horses. In this review, the principles of particulate exposure assessment, including health-related aerosol size fractions and size-selective sampling, the factors influencing air quality in equine environments, and the effect of air quality on the equine respiratory tract are discussed. The objective of this review is to provide the reader with a summary of the most common chronic inflammatory airway diseases in the horse and the principles of air sampling that are essential to the planning, interpretation, and assessment of equine respiratory health-related exposure studies.

Environmental exposures and airway inflammation in young thoroughbred horses.

BACKGROUND: Inflammatory airway disease (IAD) in horses is a widespread, performance-limiting syndrome believed to develop in response to inhaled irritants in the barn environment. OBJECTIVES: To evaluate changes in bronchoalveolar lavage fluid (BALF) cytology and exposure to particulates, endotoxin, and ammonia during horses' first month in training. ANIMALS: Forty-nine client-owned 12- to 36-month-old Thoroughbred horses entering race training. METHODS: In this prospective cohort study, a convenience sample of horses was assigned to be fed hay from a net (n = 16), whereas the remaining horses were fed hay from the ground (n = 33). BALF was collected at enrollment and after 14 and 28 days in training. Respirable particulate, inhalable particulate, respirable endotoxin, and ammonia concentrations were measured at the breathing zone of each horse weekly. RESULTS: Median respirable particulates were significantly higher when horses were fed from hay nets than when fed hay from the ground (hay net 0.28 mg/m(3) , no hay net 0.055 mg/m(3) , P < .001). Likewise, inhalable particulate (hay net 8.3 mg/m(3) , no hay net 3.3 mg/m(3) , P = .0064) and respirable endotoxin (hay net 173.4 EU/m(3) , no hay net 59.2 EU/m(3) , P = .018) exposures were significantly higher when horses were fed from hay nets. Feeding hay from a net resulted in significantly higher BALF eosinophil proportions over time (P < .001). BALF eosinophils were significantly related to respirable particulate exposure (14 days in training rs = 0.37, P = .012, 28 days in training, rs = 0.38, P = .017). CONCLUSIONS AND CLINICAL IMPORTANCE: Pulmonary eosinophilic inflammation develops in response to respirable particulate exposure in young Thoroughbreds, indicating a potential hypersensitivity to inhaled particulate allergens.

Maintenance of arytenoid abduction following carbon dioxide laser debridement of the articular cartilage and joint capsule of the cricoarytenoid joint combined with prosthetic laryngoplasty in horses: an in vivo and in vitro study.

The objective was to evaluate CO2 laser debridement of the cricoarytenoid joint (CAJ) combined with prosthetic laryngoplasty to prevent post-operative loss of arytenoid abduction in seven horses. Horses were assigned to either laser debridement of the left CAJ and laryngoplasty (laser treated, n=5) or control laryngoplasty (sham, n=2), and were evaluated with endoscopic examinations and measurement of right to left angle quotients (RLQ) to assess maintenance of arytenoid abduction. The animals were euthanased at intervals after surgery and larynges were harvested for post-mortem testing, including determination of translaryngeal flow, pressure, impedance and RLQ. Measurements were obtained under increasing vacuum-generated negative pressure with laryngoplasty sutures intact and with the knot/crimp of the laryngoplasty sutures removed. Following post-mortem testing the cricoarytenoid joints were examined histologically. Post-operative endoscopic examinations revealed no significant differences between RLQ measurements calculated for day 1 following surgery to the termination date of the study for the seven horses. Post-mortem RLQ at airflows of 10 and 60 L/s was significantly higher in sham than in laser treated horses both before and after knot/crimp removal. Translaryngeal impedance at 10 and 60 L/s was not statistically different between groups. Histopathology revealed necrosis and loss of articular cartilage in the laser treated horses. The lymphoid cell infiltration subsided but joint capsule and periarticular fibrosis increased over the course of the study. Post-operative loss of arytenoid abduction after laryngoplasty can be minimized with CO2 laser debridement of the CAJ joint.

Effects of N-butylscopolammonium bromide on lung function in horses with recurrent airway obstruction.

BACKGROUND: Equine recurrent airway obstruction (RAO) is characterized by airway inflammation, bronchoconstriction, and increased mucus production in the airways. Anticholinergic drugs like atropine induce bronchodilatation and rapid improvement in lung function. N-butylscopolammonium bromide (NBB) is an anticholinergic drug used to relieve spasmodic colic in horses, but its effect on airway smooth muscle is unknown. OBJECTIVE: To evaluate the effect of NBB on clinical signs and lung function of RAO-affected horses. ANIMALS: Nine horses diagnosed with RAO. METHODS: Double-blind, placebo-controlled, randomized crossover trial. Horses were challenged with moldy hay until a maximum change in transpulmonary pressure (∆P(L) (max) ) > 15 cm H(2) O was achieved. NBB (0.3 mg/kg) or placebo (0.9% saline) was administered IV. Clinical scores and lung function were recorded at baseline and then periodically after treatment administration. Horses were allowed a 6-week washout before administration of opposite treatments. RESULTS: Clinical score at 10 and 30 minutes (8.7 ± 2.8 and 8.7 ± 3.2, respectively) after NBB administration was significantly lower than baseline (10.8 ± 2.4). NBB administration resulted in a significant decrease in ∆P(L) (max) (baseline: 35.1 ± 6.9 cm H(2) O) starting 2 minutes after administration (16.3 ± 6.6 cmH(2) O) with a maximum decline observed at 10 minutes (13.5 ± 7.1 cm H(2) O). ∆P(L) (max) values between 60 and 120 minutes after NBB administration were not different from placebo. CONCLUSION AND CLINICAL IMPORTANCE: N-butylscopolammonium bromide is a potent bronchodilator, reaching maximum effect 10 minutes after intravenous administration. This effect dissipates within 1 hour of drug administration.

Effect of sample handling and storage time on the stability of total CO2 in equine plasma.

REASONS FOR PERFORMING STUDY: The stability of total CO2 concentration (ctCO2) in plasma is influenced by storage temperature and handling during sample processing. Conflicting information exists regarding the stability of ctCO2 in equine plasma over time, and the effect of centrifugation on the measured value for plasma ctCO2 is unclear. OBJECTIVES: To determine plasma ctCO2 stability over 5 days when equine blood is collected into Vacutainer tubes, centrifuged within 30 min of collection, and stored at 4 degrees C; and to determine whether a delay in centrifugation increases the rate at which plasma ctCO2 decreases over time. METHODS: Blood was collected from 6 adult horses into 3 ml plastic Vacutainer tubes and randomly assigned to be centrifuged immediately, or after, storage. Plasma ctCO2 was measured in triplicate at 0, 24, 48, 72 and 96 h after collection using a NOVA-4 analyser Data were analysed using multivariable linear regression, with P < 0.05 being defined as significant. RESULTS: Plasma ctCO2 decreased linearly over time during storage at 4 degrees C. The measured value for ctCO2 decreased at a faster rate (-0.28 mmol/l/day; P < 0.0001) when centrifugation was delayed, compared with immediate centrifugation (-0.10 mmol/l/day). There was a significant effect of sequence of sample analysis on the ctCO2 value when measured in triplicate: the second and third measurements were 0.31 and 0.41 mmol/l lower than the first measurement, respectively. CONCLUSIONS: Blood samples collected from horses into Vacutainer tubes should be centrifuged immediately after collection and analysed as soon as possible to ensure accurate values for plasma ctCO2. POTENTIAL RELEVANCE: Failure to centrifuge, or excessive delay in measuring ctCO2 after centrifugation, produces values significantly lower than the true value.

Comparative efficacy of inhaled albuterol between two hand-held delivery devices in horses with recurrent airway obstruction.

REASONS FOR PERFORMING STUDY: Studies investigating the clinical efficacy of albuterol administered with the same propellant and commercially available delivery devices in horses with recurrent airway obstruction (RAO) are not currently available. OBJECTIVES: To determine the efficacy of aerosolised albuterol administered to horses with RAO by means of 2 commercially available, hand-held delivery devices. METHODS: Ten horses with RAO were kept in a dusty environment and fed mouldy hay to induce airway obstruction. Lung mechanics were measured before and after the procedure. ΔP(max) was measured 5 min after administration of 180 µg of albuterol from a pressurised metered dose inhaler, using an aerosol delivery device chosen randomly. This process was repeated every 5 min until maximal bronchodilation was achieved. After a 24 h washout period, lung mechanics data were again collected using the other aerosol delivery device. RESULTS: Aerosolised albuterol induced a significant and rapid bronchodilation in the horses using both aerosol delivery devices. No statistically significant difference in pulmonary function was observed in response to albuterol therapy between the 2 devices. The dose required to achieve 50% of maximal bronchodilation was not statistically different between the 2 devices (173.35 ± 78.35 µg with Device 1 and 228.49 ± 144.99 µg with Device 2, P = 0.26). The decrease in lung resistance tended to be more pronounced after albuterol administration with Device 1 (P = 0.066). CONCLUSIONS: Aerosolised albuterol is an effective bronchodilator in horses with recurrent airway obstruction. There is no statistically significant difference between the 2 commercially available aerosol delivery devices in terms of efficacy. POTENTIAL RELEVANCE: Aerosolised albuterol is effectively delivered using currently available devices leading to maximal bronchodilation in horses with RAO at an average dose of 540 µg.

A multicenter retrospective study of 151 renal biopsies in horses.

BACKGROUND: Renal biopsies are uncommonly performed in horses and little is known about their diagnostic utility and associated complication rate. OBJECTIVE: To describe the techniques, the complication rate, risk factors, and histopathology results; as well as evaluate the safety and diagnostic utility of renal biopsy in the horse. ANIMALS: One hundred and forty-six horses from which 151 renal biopsies were obtained. Animals ranged in age from 48 hours to 30 years. METHODS: Multicenter retrospective study, with participation of 14 institutions (1983-2009). RESULTS: Renal biopsy in horses was associated with a similar rate of complications (11.3%) to that occurring in humans and companion animals. Complications were generally associated with hemorrhage or signs of colic, and required treatment in 3% of cases. Fatality rate was low (1/151; 0.7%). Biopsy specimens yielded sufficient tissue for a histopathologic diagnosis in most cases (94%) but diagnoses had only fair (72%) agreement with postmortem findings. Risk factors for complications included biopsy specimens of the left kidney (P = .030), a diagnosis of neoplasia (P = .004), and low urine specific gravity (P = .030). No association with complications was found for age, sex, breed, institution, presenting complaint, other initial clinicopathologic data, biopsy instrument, needle size, or use of ultrasonographic guidance. CONCLUSIONS AND CLINICAL IMPORTANCE: Renal biopsy in horses has low morbidity and results in a morphological histopathologic diagnosis in 94% of cases. However, this procedure might result in serious complications and should only be used when information obtained would be likely to impact decisions regarding patient management and prognosis.

Efficacy and safety of sound wave treatment of recurrent airway obstruction in horses.

BACKGROUND: One proposed nonmedical therapy for recurrent airway obstruction (RAO) in horses is a handheld acoustic device that propels sound waves from the nose down the tracheobronchial tree where it is intended to dislodge mucous and relax bronchospasm, permitting clearance of mucoid secretions. OBJECTIVE: To evaluate the effectiveness and safety of this device when used as per the manufacturer's recommendations as a treatment for RAO. ANIMALS: Nine adult horses previously diagnosed with RAO. METHODS: Prospective, cross-over clinical trial. Horses were exposed to a dusty environment until airway obstruction developed as defined by standard lung mechanics (SLM). Horses were randomly assigned to receive either acoustic therapy or a sham treatment for 4 weeks while being maintained in this environment. Horses were evaluated by clinical scores, SLM, and forced expiration regularly for 4 weeks. The opposite treatment was administered after a washout period. RESULTS: Seven horses received the treatment; 9 received the sham. There were no changes (P>.05) in clinical score, maximal change in transpulmonary pressure (ΔPLmax), lung resistance (RL), or the forced expiratory flow rate averaged over the last 75-95% of expiration (FEF75-95%) over the study period. The device was determined to be safe, although several minor adverse effects were noted, including head tossing, coughing, and chewing during treatment. CONCLUSIONS AND CLINICAL IMPORTANCE: Treatment with this device did not improve clinical signs or lung function in horses with RAO kept in a dusty environment. Currently accepted treatments, including environmental management and medical therapy, should be recommended.

Effects of collecting blood into plastic heparinised vacutainer tubes and storage conditions on blood gas analysis values in horses.

REASONS FOR PERFORMING STUDY: Plastic heparinised vacutainer tubes are used for blood gas analysis in horses. This collection method may not be ideal because influx of atmospheric O(2) through the permeable plastic wall of the vacutainer tube and loss of CO(2) into the gas phase above the blood sample should increase blood PO(2) and decrease PCO(2), respectively. OBJECTIVES: To determine the effects of collecting blood into plastic vacutainer tubes and storage conditions on blood gas analysis values. METHODS: Blood was obtained from 6 healthy horses and tonometered at 37 °C with 12% O(2) and 5% CO(2). Three ml aliquots of tonometered blood were collected using a glass syringe or vacutainer tube and stored in iced water or at room temperature for 0, 5, 15, 30, 60 and 120 min. Blood samples from vacutainer tubes were collected aerobically (tube opened for 5 s) or anaerobically (tube remained closed). Blood gas analysis was performed in duplicate using a Radiometer ABL5. Data was analysed using repeated measures analysis of variance and P < 0.05 was significant. RESULTS: Compared to the glass syringe, tonometered blood collected in vacutainer tubes had an immediate, significant, sustained and marked increase in PO(2) and an immediate, significant, transient but small decrease in PCO(2). Blood PO(2) and PCO(2) were higher when vacutainer tubes were stored in iced water instead of at room temperature. Measured blood pH and calculated values for plasma bicarbonate and total CO(2) concentration and base excess of extracellular fluid were similar when blood was collected in glass syringes or vacutainer tubes and values were not altered by storage temperature or time. CONCLUSIONS: Plastic heparinised vacutainer tubes should not be used to collect samples for measurement of blood PCO(2) and PO(2). Vacutainer tubes provide an accurate method for measuring plasma bicarbonate concentration, total CO(2) concentration and base excess.

Partial divergence of cytokine mRNA expression in bronchial tissues compared to bronchoalveolar lavage cells in horses with recurrent airway obstruction.

The aim of this study was to investigate mRNA levels of cytokines in bronchial epithelium in horses with recurrent airway obstruction (RAO) during acute crisis and remission. Additionally, cytokine mRNA levels in endobronchial biopsies and bronchoalveolar lavage (BAL) cells were compared. Seven RAO horses were examined while in respiratory crisis following provocation and again while in remission after 2 months on pasture, during which time six healthy horses on pasture were also examined. Quantitative real-time PCR (RT-PCR) was used to assess mRNA expression for cytokines IL-5, IL-6, IL-8, IL-10, IL-17 and transforming growth factor beta1 (TGF-beta1) in endobronchial biopsies and bronchoalveolar lavage. Expression of IL-8 mRNA was significantly upregulated during crisis in both endobronchial biopsies and BAL cells (p=0.036), while there was a similar trend for upregulation of IL-10 mRNA only in BAL cells that approached significance (p=0.059). Moreover, during crisis the expression of IL-8 mRNA in BAL cells was positively correlated to relative IL-6 mRNA expression (r(s)=0.971, p=0.001) and bronchial epithelial expression of IL-10 and TGF-beta1 mRNA were positively correlated (r(s)=0.943, p=0.005). In comparing the relationship of mRNA expression in BAL to biopsy in individual RAO horses, there was a positive correlation with IL-6 to IL-8 mRNA expression in BAL during respiratory crisis (r(s)=0.971, p=0.001) that also correlated positively with IL-8 expression in biopsies on pasture (r(s)=0.986, p<0.0001 for both). Regarding RAO horses at pasture versus controls neither the cytokine mRNA levels in endobronchial biopsy nor in BAL cells differed significantly. These results further support previous findings that IL-8 mRNA in both BAL cells and bronchial epithelium is upregulated in RAO horses during crisis. However, apart from IL-8, it appears that expression of other cytokines, including IL-5, IL-6, IL-10, IL-17 and TGF-beta1 in bronchial epithelium does not necessarily mirror cytokine expression in BAL cells in individual horses with RAO. Accordingly, examination of markers of inflammation in endobronchial tissue provides complementary but not necessarily identical information to that obtained in BAL cells. Given the potential for repeated sampling over time bronchial biopsy can serve as an invaluable additional tool for investigation of time-dependent changes in inflammatory process in this animal model of asthma.