Evaluation of the airway mechanics of modified toggle laryngoplasty constructs using a vacuum chamber airflow model.

OBJECTIVE: To evaluate the airway mechanics of modified toggle LP constructs in an airflow chamber model and compare these to the airway mechanics of standard LP constructs. STUDY DESIGN: Ex-vivo experimental study. SAMPLE POPULATION: Fifty-one equine cadaveric larynges. METHODS: Bilateral LP constructs were performed using a modified toggle (n = 23) or a standard (n = 21) LP technique. Constructs were tested in an airflow model before and after cyclic loading which was designed to mimic postoperative swallowing. The cross-sectional area (CSA), peak translaryngeal airflow (L/s), and impedance (cmH2 0/L/s) were determined and compared between LP constructs before and after cycling. RESULTS: The mean CSA of the rima glottidis of the modified toggle LP constructs was 15.2 ± 2.6 cm2 before and 14.7 ± 2.6 cm2 after cyclic loading, and the mean CSA of the rima glottidis of the standard LP constructs was 16.4 ± 2.9 cm2 before and 15.7 ± 2.8 cm2 after cyclic loading. The modified toggle LP constructs had similar peak translaryngeal impedance before and after cyclic loading (p = .13); however, the standard LP constructs had higher peak translaryngeal impedance after cyclic loading (p = .02). CONCLUSION: The modified toggle and standard LP constructs had comparable airway mechanics in an ex-vivo model. CLINICAL SIGNIFICANCE: Further investigation is warranted to determine the extent to which the modified toggle LP technique restores normal airway function in horses with RLN.

Airway Diagnostics: Bronchoalveolar Lavage, Tracheal Wash, and Pleural Fluid.

Indications for bronchoalveolar lavage, tracheal wash, and thoracocentesis for the diagnosis of respiratory diseases are discussed. Each technique is described in detail and illustrated by videos. Sample handling, preparation and evaluation are reviewed. The advantages and limitations of bronchoalveolar lavage and tracheal wash procedures as well as a critical comparison between the 2 techniques for equine asthma diagnosis are presented. Finally, validated cut-off values for equine asthma diagnosis are reviewed.

The influence of hay steaming on clinical signs and airway immune response in severe asthmatic horses.

BACKGROUND: Avoidance of antigenic stimuli was found to significantly reverse airway obstruction of horses with severe equine asthma (sEA). To date, no published study investigated the influence of steaming hay on lower airway condition of sEA-affected horses. The objectives were to determine the clinical, cytological and cytokine respiratory responses of both sEA and control (CTL) horses experimentally exposed to steamed or dry hay. RESULTS: A cohort of 6 sEA horses and 6 CTL horses was involved in this field study. On day 0, both groups were fed with steamed hay for 5 consecutive days, followed by a wash-out period of 26 days prior to be fed with dry hay for 5 consecutive days. Investigations performed 2 days prior to and 5 days after each challenge included clinical score, tracheal mucus accumulation, and bronchoalveolar lavage fluid (BALF) cytology and cytokine mRNA expression. Feeding steamed hay significantly decreased its mould content (P < 0.001). Mucus score significantly increased when feeding dry hay (P = 0.01). No significant influence of challenge type was found on clinical score. Percentages of neutrophils (P < 0.001) as well as mRNA expression of IL-1ß (P = 0.024), IL-6R (P = 0.021), IL-18 (P = 0.009) and IL-23 (P = 0.036) in BALF of sEA affected horses were significantly increased after both (steamed and dry hay) challenges. Relative mRNA expression of IL-1ß, IL-6R and IL-23 in BALF were also significantly correlated to neutrophil percentages and both clinical and tracheal mucus score. CONCLUSIONS: Steaming significantly decreased mould content but inconsistently influenced the respiratory response of sEA affected horses when fed hay. Based on BALF cytology and cytokine profiles, its relevance might be controversial as a non-medicinal therapy for sEA-affected horses.

Microarray molecular mapping of horses with severe asthma.

BACKGROUND: Severe asthma (SA) in horses, resembling human asthma, is a prevalent, debilitating allergic respiratory condition marked by elevated allergen-specific immunoglobulin E (IgE) against environmental proteins; however, research exploring the exposome's influence on IgE profiles is currently limited but holds paramount significance for diagnostic and therapeutic developments. ANIMALS: Thirty-five sports horses were analyzed, consisting of environmentally matched samples from France (5 SA; 6 control), the United States (6 SA; 6 control), and Canada (6 SEA; 6 control). METHODS: This intentional cross-sectional study investigated the sensitization profiles of SA-affected and healthy horses via serological antigen microarray profiling. Partial least square-discriminant analysis (PLS-DA) was used to identify and rank the importance of allergens for class separation (ie, affected/non-affected) as variable influence of projection (VIP), and allergen with commonality internationally established via frequency analysis. RESULTS: PLS-DA models showed high discriminatory power in predicting SA in horses from Canada (area under the curve [AUC] 0.995) and France (AUC 0.867) but poor discriminatory power in horses from the United States (AUC 0.38). Hev b 5.0101, Cyn D, Der p 2, and Rum cr were the only shared allergens across all geographical groups. CONCLUSIONS AND CLINICAL IMPORTANCE: Microarray profiling can identify specific allergenic components associated with SA in horses, while mathematical modeling of this data can be used for disease classification, highlighting the variability of sensitization profiles between geographical locations and emphasizing the importance of local exposure to the prevalence of different allergens. Frequency scoring analysis can identify important variables that contribute to the classification of SA across different geographical regions.

Effects of low-dust forages on dust exposure, airway cytology, and plasma omega-3 concentrations in Thoroughbred racehorses: A randomized clinical trial.

BACKGROUND: Racehorses commonly develop evidence of mild asthma in response to dust exposure. Diets deficient in omega-3 polyunsaturated fatty acids (Ω-3) might exacerbate this response. HYPOTHESIS: To compare dust exposure, bronchoalveolar lavage fluid (BALF) cytology, and plasma Ω-3 and specialized pro-resolving mediators (SPM) concentrations amongst racehorses fed dry hay, steamed hay, and haylage. ANIMALS: Forty-three Thoroughbred racehorses. METHODS: Prospective clinical trial. Horses were randomly assigned to be fed dry hay, steamed hay, or haylage for 6 weeks. Measures of exposure to dust in the breathing zone were obtained twice. At baseline, week-3, and week-6, BALF cytology was examined. Plasma lipid profiles and plasma SPM concentrations were examined at baseline and week 6. Generalized linear mixed models examined the effect of forage upon dust exposure, BALF cytology, Ω-3, and SPM concentrations. RESULTS: Respirable dust was significantly higher for horses fed hay (least-square mean ± s.e.m. 0.081 ± 0.007 mg/m3 ) when compared with steamed hay (0.056 ± 0.005 mg/m3 , P = .01) or haylage (0.053 ± 0.005 mg/m3 , P < .01). At week 6, BALF neutrophil proportions in horses eating haylage (3.0% ± 0.6%) were significantly lower compared with baseline (5.1 ± 0.7, P = .04) and horses eating hay (6.3% ± 0.8%, P < .01). Plasma eicosapentaenoic acid to arachidonic acid ratios were higher in horses eating haylage for 6 weeks (0.51 ± 0.07) when compared with baseline (0.34 ± 0.05, P < .01) and horses eating steamed (0.24 ± 0.02, P < .01) or dry hay (0.25 ± 0.03, P < .01). CONCLUSIONS AND CLINICAL IMPORTANCE: Steamed hay and haylage reduce dust exposure compared with dry hay, but only haylage increased the ratio of anti-inflammatory to pro-inflammatory lipids while reducing BAL neutrophil proportions within 6 weeks.

Effect of Lavage Solution Type on Bronchoalveolar Lavage Fluid Cytology in Clinically Healthy Horses.

Equine bronchoalveolar lavage (BAL) is usually performed with 250-500 mL of isotonic saline at pH 5.5. The acidic pH of saline may cause an increase in airway neutrophil count 48 h after BAL. Other isotonic solutions such as Ringer's solution, phosphate-buffered saline (PBS) or Plasma-Lyte 148® have a neutral pH of 7.4 and might be a better choice for BAL by not provoking inflammation and the influx of neutrophils into airways. BAL was performed in four healthy horses in four different lung lobes using four different solutions in a randomized crossover design. In each lobe, BAL was performed twice with a 48 h interval using 250 mL of solution. Automated total nucleated cell counts (TNCs) were recorded, and differential cell counts in lavage fluid were determined by two investigators blinded to treatments. The mean volume of BAL fluid retrieved was 51 ± 14%. The mean neutrophil percentage (%N) increased from 1.5 ± 0.9% to 14.7 ± 9.6% at 48 h (p < 0.001) but was not significantly affected by the solution used or the lung lobe sampled. In conclusion, in this study, the influx of neutrophils into airways after BAL was independent of the type of isotonic solution used and the lung lobe sampled. Saline remains an appropriate solution for BAL in horses.

A randomised, controlled trial to determine the effect of levothyroxine on Standardbred racehorses.

BACKGROUND: The use of thyroid supplement is pervasive in athletic horses although its effects on measures of performance are not known. OBJECTIVES: One purpose of this study was to determine whether supra-physiologic doses of levothyroxine affect the velocities at which blood lactate was greater than 4 mmol/L (VLa4 ) and heart rate was over 150 (V150 ) and 200 (V200 ) beats per minute respectively. Additionally, a survey of post-race blood samples was also conducted to determine whether high thyroxine concentrations were common in racehorses. STUDY DESIGN: A randomised, crossover, trial was performed in six healthy Standardbred racehorses. METHODS: Study 1: T4 was determined in 50 post-race samples from a single Standardbred meet. Study 2: Research horses were trained to fitness and then randomised to one of three treatments: carrier, 0.1 mg/kg thyroxine or 0.25 mg/kg thyroxine for 2 weeks. Horses completed a standardised exercise treadmill test (SET) to fatigue on the last day of treatment. Serum free and total thyroxine and triiodothyronine were determined on the day of SET testing. Blood lactate and ECG data were collected during the SET at 6, 8, 10, 11, and 12 m/s and during recovery. The effect of treatment and SET on heart rate and blood lactate was examined using generalised linear mixed models. Post hoc analysis was adjusted for multiple comparisons using Tukey's Test. Data were expressed as mean ± standard deviation and P < .05 was considered significant. RESULTS: Study 1: The median T4 value in this population of horses was 2.00 µg/dL (laboratory's normal range 1.5-4.5 µg/dL) and 3 of 50 racehorses had values above the laboratory reference range. Study 2: Levothyroxine at 0.25 mg/kg resulted in higher heart rates during SET (199 ± 30, 223 ± 17 and 239 ± 9 bpm at 6, 8 and 10 m/s respectively) and recovery (144 ± 20 and 119 ± 15 at 5 and 15 min) as compared to placebo (176 ± 18, 203 ± 10 and 219 ± 6 bpm at 6, 8, and10 m/s and 126 ± 5, 102 ± 11 at 5-15 minutes respectively). Three of six horses developed cardiac arrhythmias including atrial fibrillation. MAIN LIMITATIONS: A relatively small number of animals were used and a SET is not identical to actual racing conditions. CONCLUSIONS: Supra-physiologic thyroxine supplementation caused a decreased V200 during a standard exercise test and may result in cardiac arrhythmias.

A Programmable Dual-Regime Spray for Large-Scale and Custom-Designed Electronic Textiles.

Increasing demand for wearable healthcare synergistically advances the field of electronic textiles, or e-textiles, allowing for ambulatory monitoring of vital health signals. Despite great promise, the pragmatic deployment of e-textiles in clinical practice remains challenged due to the lack of a method in producing custom-designed e-textiles at high spatial resolution across a large area. To this end, a programmable dual-regime spray that enables the direct custom writing of functional nanoparticles into arbitrary fabrics at sub-millimeter resolution over meter scale is employed. The resulting e-textiles retain the intrinsic fabric properties in terms of mechanical flexibility, water-vapor permeability, and comfort against multiple uses and laundry cycles. The e-textiles tightly fit various body sizes and shapes to support the high-fidelity recording of physiological and electrophysiological signals on the skin under ambulatory conditions. Pilot field tests in a remote health-monitoring setting with a large animal, such as a horse, demonstrate the scalability and utility of the e-textiles beyond conventional devices. This approach will be suitable for the rapid prototyping of custom e-textiles tailored to meet various clinical needs.

Lipidomic analysis of surfactant and plasma from horses with asthma and age-matched healthy horses.

OBJECTIVE: To perform lipidomic analysis of surfactant and plasma from asthmatic and healthy horses. ANIMALS: 30 horses with clinical signs of asthma and 30 age-matched control horses. PROCEDURES: Detailed history, physical examination, CBC, and bronchoalveolar lavage fluid (BALF) cytologies were obtained. Asthmatic horses were grouped based on their BALF inflammatory profile: severe equine asthma (SEA), mild equine asthma with neutrophilic airway inflammation (MEA-N), or mild equine asthma with eosinophilic airway inflammation (MEA-E). Each asthma group was assigned its own age-matched control group. Lipidomic analysis was completed on surfactant and plasma. Surfactant protein D (SP-D) concentrations were measured in serum and BALF. RESULTS: SEA surfactant was characterized by a phospholipid deficit and altered composition (increased ceramides, decreased phosphatidylglycerol, and increased cyclic phosphatidic acid [cPA]). In comparison, MEA-N surfactant only had a decrease in select phosphatidylglycerol species and increased cPA levels. The plasma lipidomic profile was significantly different in all asthma groups compared to controls. Specifically, all groups had increased plasma phytoceramide. SEA horses had increased plasma cPA and diacylglycerol whereas MEA-N horses only had increased cPA. MEA-E horses had increases in select ceramides and dihydrocermides. Only SEA horses had significantly increased serum SP-D concentrations. CLINICAL RELEVANCE: The most significant surfactant alterations were present in SEA (altered phospholipid content and composition); only mild changes were observed in MEA-N horses. The plasma lipidomic profile was significantly altered in all groups of asthmatic horses and differed among groups. Data from a larger population of asthmatic horses are needed to assess implications for diagnosis, prognosis, and treatment.

Comparison of genomic and proteomic data in recurrent airway obstruction affected horses using Ingenuity Pathway Analysis®.

BACKGROUND: Recurrent airway obstruction (RAO) is a severe chronic respiratory disease affecting horses worldwide, though mostly in the Northern hemisphere. Environmental as well as genetic factors strongly influence the course and prognosis of the disease. Research has been focused on characterization of immunologic factors contributing to inflammatory responses, on genetic linkage analysis, and, more recently, on proteomic analysis of airway secretions from affected horses. The goal of this study was to investigate the interactions between eight candidate genes previously identified in a genetic linkage study and proteins expressed in bronchoalveolar lavage fluid (BALF) collected from healthy and RAO-affected horses. The analysis was carried out with Ingenuity Pathway Analysis® bioinformatics software. RESULTS: The gene with the greatest number of indirect interactions with the set of proteins identified is Interleukin 4 Receptor (IL-4R), whose protein has also been detected in BALF. Interleukin 21 receptor and chemokine (C-C motif) ligand 24 also showed a large number of interactions with the group of detected proteins. Protein products of other genes like that of SOCS5, revealed direct interactions with the IL-4R protein. The interacting proteins NOD2, RPS6KA5 and FOXP3 found in several pathways are reported regulators of the NFκB pathway. CONCLUSIONS: The pathways generated with IL-4R highlight possible important intracellular signaling cascades implicating, for instance, NFκB. Furthermore, the proposed interaction between SOCS5 and IL-4R could explain how different genes can lead to identical clinical RAO phenotypes, as observed in two Swiss Warmblood half sibling families because these proteins interact upstream of an important cascade where they may act as a functional unit.

Lung volume reduction by bronchoscopic administration of steam.

RATIONALE: At present, bronchoscopic approaches to lung volume reduction (LVR) create airway obstruction to achieve parenchyma collapse, avoiding many risks of surgical LVR. However, LVR by these methods is limited by temporary or incomplete collapse and/or residual atelectatic and scarred tissue volumes. Heat-induced ablation of lung tissue is currently under investigation as an alternative LVR methodology. OBJECTIVES: We hypothesized that bronchoscopic steam injection can produce safe and effective LVR, and explored potential mechanisms for the effects. METHODS: Steam treatments were applied bilaterally to six cranial lobe segments of large dogs. For series 1, 14 dogs received one of three target heat dose levels (1, 4, or 8 cal · ml⁻¹ segment volume), and then 3 months of follow-up including pulmonary function testing and monitoring for complications. For series 2, 12 dogs received a single target dose (4 cal · ml⁻¹) or sham, similar follow-up, and then assessment of lobar mass, volume, and blood flow. Vapor content of series 2 steam was 40% greater than for series 1 (similar heat dose) to attempt more peripheral heat delivery. MEASUREMENTS AND MAIN RESULTS: Nineteen of 20 treatment animals survived with minimal evidence of serious risks or reduced pulmonary function testing volumes, but 1 died from pneumothorax 5 days post-treatment. Postmortem processing of animals that survived as planned revealed obvious dose-dependent lobe reductions, additional evidence of risks, and blood flow reduction that occurred immediately post-treatment. CONCLUSIONS: Bronchoscopic administration of steam is a potentially safe means to achieve LVR, but substantial risks are present and further research is recommended.

Equine respiratory viruses, airway inflammation and performance in thoroughbred racehorses.

Equine asthma is a common cause of poor performance in racehorses but it is unclear if respiratory viruses contribute to its etiology. The objective of the study was to determine if respiratory viruses were associated with clinical signs and bronchoalveolar lavage fluid (BALF) cytology in Thoroughbred racehorses. Equine herpesviruses (EHV-1, 2, 4, 5) and equine rhinitis A and B viruses (ERBV, ERAV) genomes were quantified by qPCR in nasopharyngeal, tracheal, and BALF samples collected after racing. The relationships between virus detection and load and clinical signs, performance, BALF cytology, and environmental exposures were examined with generalized linear mixed models. Ninety-two samples were collected from 31 horses. EHV-1 and ERAV were not found; EHV-4 was detected in only one sample. EHV-2, EHV-5 and ERBV were more likely to be detected in upper airway samples than in BALF (P < 0.0001). Neither respiratory virus detection nor load was associated with clinical signs or performance. Nasopharyngeal detection and load of ERBV and tracheal detection and load of EHV-5 were associated with increased proportions of neutrophils in BALF (P < 0.003). However, nasopharyngeal detection and load of EHV-5 was not (P = 0.11). Nasopharyngeal detection and load of EHV-2 were associated with decreased BALF mast cell proportions. Respirable dust exposures were significantly higher in horses with detection of ERBV when compared to horses with no detectable ERBV (P < 0.001). Our results suggest that ERBV, EHV-2 and EHV-5 are commonly present in upper airways of healthy racehorses; however, the role they play in the etiology of equine asthma remains unclear.

Effects of forages, dust exposure and proresolving lipids on airway inflammation in horses.

OBJECTIVE: To investigate the role of omega-3 polyunsaturated fatty acids (Ω-3)-derived proresolving lipid mediators (PRLM) in the resolution of mild airway inflammation in horses. ANIMALS: 20 horses with mild airway inflammation. PROCEDURES: Horses previously eating hay were fed hay pellets (low Ω-3 content; n = 10) or haylage (high Ω-3 content; 9) for 6 weeks. Dust exposure was measured in the breathing zone with a real-time particulate monitor. Bronchoalveolar lavage (BAL) was performed at baseline, week 3, and week 6. The effect of PRLM on neutrophil apoptosis and efferocytosis was examined in vitro. BAL fluid inflammatory cell proportions, apoptosis of circulating neutrophils, efferocytosis displayed by alveolar macrophages, and plasma lipid concentrations were compared between groups fed low and high amounts of Ω-3 by use of repeated measures of generalized linear models. RESULTS: Dust exposure was significantly higher with hay feeding, compared to haylage and pellets, and equivalent between haylage and pellets. BAL fluid neutrophil proportions decreased significantly in horses fed haylage (baseline, 11.8 ± 2.4%; week 6, 2.5 ± 1.1%) but not pellets (baseline, 12.1 ± 2.3%; week 6, 8.5% ± 1.7%). At week 6, horses eating haylage had significantly lower BAL neutrophil proportions than those eating pellets, and a significantly lower concentration of stearic acid than at baseline. PRLM treatments did not affect neutrophil apoptosis or efferocytosis. CLINICAL RELEVANCE: Despite similar reduction in dust exposure, horses fed haylage displayed greater resolution of airway inflammation than those fed pellets. This improvement was not associated with increased plasma Ω-3 concentrations. Feeding haylage improves airway inflammation beyond that due to reduced dust exposure, though the mechanism remains unclear.

The effect of lower airway inflammation on inflammatory cytokine gene expression in bronchoalveolar lavage fluid and whole blood in racing Thoroughbreds.

BACKGROUND: Immunological mechanisms involved in the pathogenesis of mild to moderate equine asthma (MEA) are not completely understood. There are limited data on bronchoalveolar lavage fluid (BALF) and blood inflammatory cytokine profiles in racehorses with MEA, and the effect of racing on inflammatory cytokines is unknown. HYPOTHESIS/OBJECTIVES: We hypothesized that inflammatory cytokine gene expression in BALF and resting blood would be higher in racehorses with lower airway inflammation compared to healthy controls, and that gene expression in blood collected immediately post-race would be increased compared to resting blood in racehorses with lower airway inflammation. ANIMALS: 38 racing Thoroughbreds (samples: 30 resting blood, 22 post-race BALF, 41 post-race blood). METHODS: Prospective observational study. Inflammatory cytokine gene expression was determined in resting blood, post-race BALF and post-race blood from racehorses with lower airway inflammation and controls. RESULTS: Lower airway inflammation was diagnosed in 79 % of racehorses (23 % neutrophilic, 67 % mastocytic, and 10 % mixed). There was no difference in gene expression in BALF or resting blood between racehorses with lower airway inflammation and controls. IL-8 gene expression was higher in post-race blood compared to resting peripheral blood, regardless of disease (p = 0052). BALF neutrophil proportions increased with increasing IL-1ß gene expression in all sample types (p = 0.0025). BALF mast cell proportions increased with increasing TNF-α gene expression in post-race blood (p = 0.015). CONCLUSIONS AND CLINICAL IMPORTANCE: Lower airway inflammation was common in a population of racehorses without respiratory signs or exercise intolerance. Exercise alone increased peripheral blood IL-8 gene expression. Inflammatory cytokine gene expression was not increased in BALF or resting blood in horses with subclinical lower airway inflammation, precluding its diagnostic utility in clinical practice.

Evaluation of the effects of transendoscopic diode laser palatoplasty on clinical, histologic, magnetic resonance imaging, and biomechanical findings in horses.

OBJECTIVE: To determine the effects of diode laser palatoplasty on the soft palate in horses. ANIMALS: 6 clinically normal horses and 6 euthanized horses from another study. PROCEDURES: 6 horses underwent diode laser palatoplasty (treated horses); 3 received low-dose laser treatment (1,209 to 1,224 J), and 3 received high-dose treatment (2,302 to 2,420 J). Six other horses received no treatment (control horses). The upper respiratory tracts of all treated horses were evaluated immediately following surgery (day 0) and on days 2, 7, 14, 21, 30, and 45. Horses were euthanized on day 45, and magnetic resonance imaging (MRI) of the head was performed. The soft palate was removed from treated and control horses, evaluated grossly, and scored for edema, inflammation, and scarring. Soft palates from all horses were sectioned for histologic and biomechanical evaluations. RESULTS: Endoscopic examination revealed a significant increase in soft palate scarring and decrease in edema and inflammation in treated horses by day 7. Gross postmortem findings corresponded with MRI findings. Gross and histologic examination revealed a significant increase in scarring, edema, and inflammation at day 45. Histologic evaluation of palatal tissue from high-dose-treated horses revealed full-thickness injury of skeletal muscle, with atrophy of muscle fibers; findings in low-dose-treated horses indicated superficial injury to skeletal muscle. After surgery, treated horses had a significant decrease in soft palate elastic modulus, compared with control horses. CONCLUSIONS AND CLINICAL RELEVANCE: Laser palatoplasty resulted in soft palate fibrosis and skeletal muscle loss; however, the fibrosis did not result in an increase in soft palate elastic modulus.

Equine Asthma: Current Understanding and Future Directions.

The 2019 Havemeyer Workshop brought together researchers and clinicians to discuss the latest information on Equine Asthma and provide future research directions. Current clinical and molecular asthma phenotypes and endotypes in humans were discussed and compared to asthma phenotypes in horses. The role of infectious and non-infectious causes of equine asthma, genetic factors and proposed disease pathophysiology were reviewed. Diagnostic limitations were evident by the limited number of tests and biomarkers available to field practitioners. The participants emphasized the need for more accessible, standardized diagnostics that would help identify specific phenotypes and endotypes in order to create more targeted treatments or management strategies. One important outcome of the workshop was the creation of the Equine Asthma Group that will facilitate communication between veterinary practice and research communities through published and easily accessible guidelines and foster research collaboration.

Effect of furosemide and furosemide-carbazochrome combination on exercise-induced pulmonary hemorrhage in Standardbred racehorses.

The objective was to quantify the effect of furosemide and carbazochrome on exercise-induced pulmonary hemorrhage (EIPH) in Standardbred horses using red blood cell count and hemoglobin concentration in bronchoalveolar lavage (BAL) fluid. Six healthy Standardbred horses with prior evidence of EIPH performed a standardized treadmill test 4 h after administration of placebo, furosemide, or furosemide-carbazochrome combination. Red blood cell (RBC) counts and hemoglobin concentrations were determined on the BAL fluid. The RBC count in BAL ranges were (2903-26,025 cells/microL), (45-24,060 cells/microL), and (905-3045 cells/microL) for placebo, furosemide, and furosemide-carbazochrome, respectively. Hemoglobin concentration ranges were (0.03-0.59 mg/mL), (0.01-0.55 mg/mL), and (0.007-0.16 mg/mL) for placebo, furosemide, and furosemide-carbazochrome groups, respectively. No significant differences were detected among treatments. However, there was great variability among horses, suggesting that a larger sample size or better selection of horses was needed.

Development of a comprehensive protein microarray for immunoglobulin E profiling in horses with severe asthma.

BACKGROUND: Severe asthma in horses, known as severe equine asthma (SEA), is a prevalent, performance-limiting disease associated with increased allergen-specific immunoglobulin E (IgE) against a range of environmental aeroallergens. OBJECTIVE: To develop a protein microarray platform to profile IgE against a range of proven and novel environmental proteins in SEA-affected horses. ANIMALS: Six SEA-affected and 6 clinically healthy Warmblood performance horses. METHODS: Developed a protein microarray (n = 384) using protein extracts and purified proteins from a large number of families including pollen, bacteria, fungi, and arthropods associated with the horses, environment. Conditions were optimized and assessed for printing, incubation, immunolabeling, biological fluid source, concentration techniques, reproducibility, and specificity. RESULTS: This method identified a number of novel allergens, while also identifying an association between SEA and pollen sensitization. Immunolabeling methods confirmed the accuracy of a commercially available mouse anti-horse IgE 3H10 source (R2 = 0.91). Biological fluid source evaluation indicated that sera and bronchoalveolar lavage fluid (BALF) yielded the same specific IgE profile (average R2 = 0.75). Amicon centrifugal filters were found to be the most efficient technique for concentrating BALF for IgE analysis at 40-fold. Overnight incubation maintained the same sensitization profile while increasing sensitivity. Reproducibility was demonstrated (R2 = 0.97), as was specificity using protein inhibition assays. Arthropods, fungi, and pollens showed the greatest discrimination for SEA. CONCLUSIONS AND CLINICAL IMPORTANCE: We have established that protein microarrays can be used for large-scale IgE mapping of allergens associated with the environment of horses. This technology provides a sound platform for specific diagnosis, management, and treatment of SEA.

Epithelial expression of mRNA and protein for IL-6, IL-10 and TNF-alpha in endobronchial biopsies in horses with recurrent airway obstruction.

BACKGROUND: The aim of this study was to evaluate the contribution of bronchial epithelium to airway inflammation, with focus on mRNA and protein expression of cytokines of innate immunity IL-6, IL-10 and TNF-alpha, in horses with Recurrent Airway Obstruction (RAO) during exacerbation and in remission. RESULTS: Despite marked clinical and physiologic alterations between exacerbation and after remission in the RAO horses no differences were detected in either cytokine mRNA or protein levels. Moreover, the expression of investigated cytokines in RAO horses on pasture did not differ from controls. In comparing real-time PCR analysis to results of immunohistochemistry only IL-10 mRNA and protein levels in RAO horses on pasture were significantly correlated (rs = 0.893, p = 0.007). Curiously, in controls examined on pasture the TNF-alpha protein level was positively correlated to IL-10 mRNA expression (rs = 0.967, p = 0.007) and negatively correlated to IL-6 mRNA expression (rs = -0.971, p = 0.001). CONCLUSION: Given the complementary relationship of assessing cytokines directly by immunohistochemistry, or indirectly by PCR to mRNA, the lack of significant changes in either mRNA or protein levels of IL-6, IL-10 or TNF-alpha mRNA in RAO horses in exacerbation suggests that these particular cytokines in bronchial tissue may not play a substantive role in the active inflammation of this disease. To support this contention further studies examining time dependency of expression of IL-6, IL-10 or TNF-alpha are needed, as is expansion of the range of cytokines to include other key regulators of airway inflammation.

Cholangiectasis and duodenal diverticulosis in a farm-raised elk (Cervus elaphus).

A 6-year-old male elk presented in August with a 4-month history of weight loss and a 1-day history of being recumbent and unable to rise. The elk's body condition score was 1.5/5. Hematology and blood chemistry showed an inflammatory leukogram, mild electrolyte abnormalities, and elevated liver enzyme activities. Because of poor prognosis, the owner elected euthanasia. On necropsy, 8 diverticula were in the duodenum distal to the common bile duct, which was dilated and filled with feed material. Many intrahepatic bile ducts were dilated up to 5 cm, were filled with ingesta and gritty material, and had walls thickened up to 1.5 mm. On histologic examination, mural thickening of dilated bile ducts was because of increased fibrous connective tissue with inflammatory cell infiltration and hyperplastic and hypertrophic epithelial lining. Portal tracts diffusely contained hyperplastic bile ducts and had increased amounts of fibrous connective tissue. Abscesses that contained bacteria were scattered throughout the liver. Duodenal diverticula lacked the tunica muscularis, and the muscularis mucosa was hypertrophied. Caudal to some diverticula, the duodenal lumen was narrowed. An additional finding was embolic pneumonia with fungi morphologically and immunohistochemically consistent with Aspergillus spp.