Post-exercise endoscopic and cytologic diagnosis of equine asthma syndrome in asymptomatic Brazilian pacers

The objectives were: 1. to investigate the occurrence of equine asthma syndrome (EAS) in a group of Campolina and Mangalarga Marchador horses through post-exercise respiratory endoscopy and cytology; 2. compare the efficiency of different diagnostic criteria; and 3. compare the incidence of EAS by breed, age group and sex. A standardised exercise test was conducted in the field, followed by respiratory endoscopy and bronchoalveolar lavage and fluid collection 30 min post-exercise. Clinical evaluation pre- and post-exercise, respiratory endoscopy post-exercise and bronchoalveolar lavage, also post-exercise, were conducted in 30 horses (15 Campolina and 15 Mangalarga Marchador) aged 3–18 years, without a history of respiratory illness and without clinical evidence of active respiratory disease. All horses were in active training and were considered fit to participate in equestrian competition characteristic of these breeds. Post-collection, the horses were subdivided into healthy or EAS-positive, depending on several diagnostic criteria. Statistical comparisons between breeds, age brackets (up to 7 years of age and above 7 years of age) and sex were conducted using appropriate statistical tests. According to bronchoalveolar lavage fluid (BALF) cytology, 56.7% of the horses showed at least one inclusion criteria that contributed to classification as EAS-positive. Of those, only one was classified as moderate EAS, while the others were classified as mild. Total cell count in the BALF above 530 cells/µL was the single most common diagnostic criteria (100%), followed by increased tracheal mucus (53%) at endoscopy, eosinophilia (47%) and neutrophilia (35%) in BALF. Significant differences were found between macrophage and lymphocyte numbers according to breed, but no other differences between breeds, age brackets, sex or status (healthy vs. EAS-positive). A high occurrence of EAS was found in competition-ready Mangalarga and Campolina pacer horses in this study.     

Effects of a MAPK p38 inhibitor on lung function and airway inflammation in equine recurrent airway obstruction

Reasons for performing study: It has been suggested that many of the beneficial effects of corticosteroids are mediated through mitogen‐activated protein kinase (MAPK) p38 inhibition. Objective: To investigate the efficacy of the MAPK p38 inhibitor compound MRL‐EQ1 to either prevent (Phase 1) or treat (Phase 2) recurrent airway obstruction (RAO) in horses. Methods: MRL‐EQ1 was administered i.v. at a dosage of 0.75‐1.5 mg/kg bwt q. 12 h. In Phase 1, susceptible horses in clinical remission were divided into 2 groups (n = 5/group), based on historical values of respiratory mechanics. All horses were entered in the study in pairs (one control, one treated horse) and exposed to the same environmental challenge (stabling, mouldy hay and dusty conditions). The treatment group received MRL‐EQ1 for 14 days while the control horses were untreated during the same period. In Phase 2, affected horses were ranked by severity of respiratory dysfunction and split randomly into either dexamethasone or MRL‐EQ1 treatment groups (n = 5/group). Bronchoalveolar lavage fluid, respiratory mechanic measurements, MRL‐EQ1 plasma concentration and tumour necrosis factor (TNF) whole blood activity were evaluated sequentially. Results: In Phase 1, MRL‐EQ1 did not prevent the occurrence of clinical signs and pulmonary inflammation. However, treatment was associated with a reduction in severity and a delay in the onset of signs and a reduction in pulmonary neutrophilia. In Phase 2, plasma concentrations achieved resulted in ex vivo suppression of lipopolysaccharide‐induced TNF production in equine blood. MRL‐EQ1 did not improve airway inflammation or lung function and was associated in a dose dependent manner with behavioural (depression, excitability) and blood changes (neutrophilia, increased serum muscle enzyme concentrations). Conclusions: Inhibition of p38 in the horse was partially effective in reducing clinical signs and airway inflammation when administered prior to, but not during clinical exacerbation in RAO. Potential relevance: Inhibitors of p38 MAPK with a better toxicity profile may be effective in the prevention or treatment of RAO.     

Owner assessment in judging the efficacy of airway disease treatment

Reasons for performing study: Efficacy of medications for recurrent airway obstruction is typically tested using clinical, cytological and lung function examinations of severely affected animals. These trials are technically challenging and may not adequately reflect the spectrum of disease and owner complaints encountered in clinical practice. Objective: To determine if owners of horses with chronic airway disease are better able to detect drug efficacy than a veterinarian who clinically examines horses infrequently. Method: In a double‐blinded randomised controlled trial, owners and a veterinarian compared the efficacy of dexamethasone (0.1 mg/kg bwt per os, q. 24 h, for 3 weeks; n = 9) to placebo (n = 8) in horses with chronic airway disease. Before and after treatment, owners scored performance, breathing effort, coughing and nasal discharge using a visual analogue scale (VAS). The clinician recorded vital parameters, respiratory distress, auscultation findings, cough and nasal discharge, airway mucus score, bronchoalveolar lavage fluid (BALF) cytology and arterial blood gases. Results: The VAS score improved significantly in dexamethasone‐ but not placebo‐treated horses. In contrast, the clinician failed to differentiate between dexamethasone‐ and placebo‐treated animals based on clinical observations, BALF cytology or endoscopic mucus score. Respiratory rate (RR) and arterial oxygen pressure (PaO₂) improved with dexamethasone but not placebo. Conclusions and clinical relevance: In the design of clinical trials of airway disease treatments, more emphasis should be placed on owner‐assessed VAS than on clinical, cytological and endoscopic observations made during brief examinations by a veterinarian. Quantifiable indicators reflecting lung function such as RR and PaO₂ provide a good assessment of drug efficacy.     

Respiratory diseases and their effects on respiratory function and exercise capacity

Given that aerobic metabolism is the predominant energy pathway for most sports, the respiratory system can be a rate‐limiting factor in the exercise capacity of fit and healthy horses. Consequently, respiratory diseases, even in mild forms, are potentially deleterious to any athletic performance. The functional impairment associated with a respiratory condition depends on the degree of severity of the disease and the equestrian discipline involved. Respiratory abnormalities generally result in an increase in respiratory impedance and work of breathing and a reduced level of ventilation that can be detected objectively by deterioration in breathing mechanics and arterial blood gas tensions and/or lactataemia. The overall prevalence of airway diseases is comparatively high in equine athletes and may affect the upper airways, lower airways or both. Diseases of the airways have been associated with a wide variety of anatomical and/or inflammatory conditions. In some instances, the diagnosis is challenging because conditions can be subclinical in horses at rest and become clinically relevant only during exercise. In such cases, an exercise test may be warranted in the evaluation of the patient. The design of the exercise test is critical to inducing the clinical signs of the problem and establishing an accurate diagnosis. Additional diagnostic techniques, such as airway sampling, can be valuable in the diagnosis of subclinical lower airway problems that have the capacity to impair performance. As all these techniques become more widely used in practice, they should inevitably enhance veterinarians' diagnostic capabilities and improve their assessment of treatment effectiveness and the long‐term management of equine athletes.     

Assessment of dynamic upper respiratory tract function in the equine athlete

Dynamic upper respiratory tract (URT) obstructions occur commonly in the equine athlete. During the past 25 years, veterinary diagnostic capabilities have improved greatly with the ability to perform endoscopy during exercise and thus to visualise upper airway function during various types of exercise. After correctly reproducing and diagnosing an URT obstruction, the veterinary surgeon must consider the functional significance of the endoscopic findings for that individual athlete. This is the third paper in this series and focuses on the diagnosis of URT obstructions, their relationship to the exercise test and their functional significance.     

An hypothesis on the etio-pathogenesis of equine inflammatory joint disease

The role of oxygen-derived free radicals is considered critical to the etio-pathogenesis of equine inflammatory joint disease. In vivo, the superoxide radical in the joint may be derived either from activated polymorphonuclear leukocytes or from an ischemia/reperfusion cycle. In the presence of ferrous iron, it may generate the highly reactive hydroxyl radical (OH *). Predisposing factors may include synovitis, exercise-induced ischemia and minor traumatic injury to the joints. Unlike other inflammatory mediators, oxygen-derived free radicals may damage tissue directly and these reactive species could account for the tissue injury and insidious onset of equine exercise-induced degenerative joint disease.     

Viruses associated with outbreaks of equine respiratory disease in New Zealand

AIM: To identify viruses associated with respiratory disease in young horses in New Zealand.

METHODS: Nasal swabs and blood samples were collected from 45 foals or horses from five separate outbreaks of respiratory disease that occurred in New Zealand in 1996, and from 37 yearlings at the time of the annual yearling sales in January that same year. Virus isolation from nasal swabs and peripheral blood leukocytes (PBL) was undertaken and serum samples were tested for antibodies against equine herpesviruses (EHV-1, EHV-2, EHV-4 and EHV-5), equine rhinitis-A virus (ERAV), equine rhinitis-B virus (ERBV), equine adenovirus 1 (EAdV-1), equine arteritis virus (EAV), reovirus 3 and parainfluenza virus type 3 (PIV3).

RESULTS: Viruses were isolated from 24/94 (26%) nasal swab samples and from 77/80 (96%) PBL samples collected from both healthy horses and horses showing clinical signs of respiratory disease. All isolates were identified as EHV-2, EHV-4, EHV-5 or untyped EHV. Of the horses and foals tested, 59/82 (72%) were positive for EHV-1 and/or EHV-4 serum neutralising (SN) antibody on at least one sampling occasion, 52/82 (63%) for EHV-1-specific antibody tested by enzyme-linked immunosorbent assay (ELISA), 10/80 (13%) for ERAV SN antibody, 60/80 (75%) for ERBV SN antibody, and 42/80 (53%) for haemagglutination inhibition (HI) antibody to EAdV-1. None of the 64 serum samples tested were positive for antibodies to EAV, reovirus 3 or PIV3. Evidence of infection with all viruses tested was detected in both healthy horses and in horses showing clinical signs of respiratory disease. Recent EHV-2 infection was associated with the development of signs of respiratory disease among yearlings [relative risk (RR)=2.67, 95% CI=1.59-4.47, p=0.017].

CONCLUSIONS: Of the equine respiratory viruses detected in horses in New Zealand during this study, EHV-2 was most likely to be associated with respiratory disease. However, factors other than viral infection are probably important in the development of clinical signs of disease.     

Serum concentration of surfactant protein D in horses with lower airway inflammation

Reasons for performing study: Surfactant protein D (SP‐D), mainly synthesised by alveolar type II cells and nonciliated bronchiolar cells, is one important component of innate pulmonary immunity. In man, circulating concentrations of SP‐D are routinely used as biomarkers for pulmonary injury. To date, serum SP‐D levels have only been investigated in horses in an experimental model of bacterial airway infection. Objectives: To compare serum SP‐D concentrations at rest and after exercise in horses with and without inflammatory airway disease (IAD). Methods: Venous blood samples were collected from 42 Standardbred racehorses at rest and 60 min after performing a standardised treadmill exercise test. Tracheal wash and bronchoalveolar lavage fluid (BALF) samples were collected after exercise. Based on BALF cytology, 22 horses were defined as IAD‐affected and 20 classified as controls. Serum SP‐D concentrations were assessed using a commercially available ELISA kit and statistically compared between groups of horses and sampling times. Results: Serum concentrations of SP‐D in IAD‐affected horses were significantly higher than those of control horses, both at rest and after exercise. Within the IAD‐affected group, no significant correlation was found between serum SP‐D concentrations and BALF cytology. Within each group of horses (IAD and control), no significant influence of exercise was found on serum SP‐D levels. Conclusions: This is the first study determining serum SP‐D concentrations in a noninfectious, naturally occurring form of lower airway inflammation in horses. The results highlight that IAD is associated with a detectable, though moderate, increase of circulating SP‐D levels. Potential relevance: Serum concentration of surfactant protein D could represent a potentially valuable and readily accessible blood biomarker of equine lower airway inflammation.     

Bronchoalveolar lavage fluid in Standardbred racehorses: Influence of unilateral/bilateral profiles and cut-off values on lower airway disease diagnosis

The aim of this study was to determine whether the lung side being sampled would significantly influence bronchoalveolar lavage (BAL) cytological profiles and subsequent diagnosis in Standardbred racehorses. One hundred and thirty-eight French Trotters in active training and racing were included in a prospective observational study. BAL was performed using videoendoscopy in both right and left lungs during summer meetings in 2011 (64 horses) and 2012 (74 horses). Cytological data performed 24 h later from right and left lungs were compared and specifically used to classify horses as affected with exercise-induced pulmonary haemorrhage (EIPH), inflammatory airway disease (IAD), or were ‘controls’. For IAD, cytological definition was based on two different cut off values.Neutrophil percentages, haemosiderophage percentages and the haemosiderophage/macrophage (H/M) ratios were significantly higher in the right compared to the left lung. Measures of intra-class correlation coefficients revealed a fair agreement between left and right lungs for percentages of mast cells, eosinophils, and for the H/M ratio, and a moderate agreement for neutrophil percentages. Fair to moderate agreements were observed between left and right lungs for the diagnosis of IAD and/or EIPH based on kappa coefficients. When sampling one lung only, the risk of incorrectly classifying a horse as a ‘control’ increased with the use of the restraint cut-off values for IAD. As BAL from one lung is not representative of the other lung in the same horse, both lungs should be sampled for a better assessment of lung cellularity and for a precise diagnosis of lower airway diseases.     

Lower respiratory tract disease in Thoroughbred racehorses: analysis of endoscopic data from a UK training yard

REASON FOR PERFORMING STUDY: There has been much research directed at potential causative agents and the epidemiology of lower respiratory tract disease (LRTD), but few reports of the clinical progression and outcome. OBJECTIVES: To define clinical features of LRTD in racehorses, including association with age and average duration of disease, through the analysis of endoscopic data. METHODS: A retrospective analysis of records from a single training yard of endoscopic examinations undertaken over a 2-year period was performed. Horses were subjected to regular endoscopic examination for a variety of reasons, and all horses placed on antibiotic therapy for lower respiratory tract disease were rescoped following treatment. Data analysed included a 0-8 tracheal mucus score based on visible endoscopic mucus and gross tracheal lavage turbidity, as well as age and treatment duration and type. RESULTS: A total of 522 endoscopic examinations undertaken on 123 horses and tracking 169 episodes of lower respiratory tract disease were recorded. Mean duration of disease episode was 15.5 days (median = 11 days, range = 4-61 days). Horses age 2 years were significantly more likely than those age > or=3 years to have at least one episode of respiratory disease (P<0.001). There was a direct association between initial tracheal mucus score and rescope score after treatment. CONCLUSIONS: Lower respiratory tract disease was more common in 2-year-olds than in older horses. Affected horses had endoscopic evidence of increased tracheal mucus accumulation for an average of 15.5 days per episode, a considerably shorter period than that suggested by previous studies. POTENTIAL RELEVANCE: Endoscopic examinations permit longitudinal tracking of lower respiratory tract disease in Thoroughbred racehorses. A prolonged duration of disease, sometimes extending for months, can be expected in a small proportion of cases, some of which appear to be refractory to treatment. There is a clear need for evidence-based analysis of treatment regimes to assist clinicians in decision making when managing disease in both individual and group situations.     

Inflammatory airway disease of horses

The purpose of this consensus statement is to provide a review of current knowledge and opinions concerning inflammatory airway disease (IAD) and to help practitioners differentiate IAD from heaves (or recurrent airway obstruction; RAO) and other inflammatory respiratory diseases of horses.     

Pulmonary function,airway cytology and bronchoalveolar lavage fluid drug concentration after aerosol administration of cefquinome to horses

The administration of antibiotics by aerosol to horses suffering from respiratory infections may partially circumvent the limitations of antimicrobial therapy, e.g. large injection volumes, low bioavailability and risk of diarrhoea. Only injectable formulations are available currently and usually contain other substances that could irritate the mucosa and induce coughing and bronchospasm. In addition, the quality of the aerosol, particularly in terms of the delivery of antibiotics to the deep parts of the lung, is unknown. Although used under field conditions, cefquinome delivered by aerosol has never been studied in horses. This study examined the safety of cefquinome injectable solution, administered by aerosol at a dose of 225 mg/inhalation to 7 healthy horses, by assessing (1) pulmonary function before and 15 min after a single inhalation, at the first day (Day 1) and the fifth day (Day 5) of a 5 day period treatment; and (2) the inflammatory status of the lung, i.e. percentage neutrophils and myeloperoxidase concentration, based on bronchoalveolar lavage (BAL) at D1 and D5. In addition, cefquinome concentrations were measured in bronchoalveolar lavage fluid after aerosol, intravenous (i.v.) and intramuscular (i.m.) administrations. A single aerosol of cefquinome injectable solution did not induce any immediate nor delayed pulmonary side effects in healthy horses and produced cefquinome concentrations in bronchoalveolar lavage (BAL) within 30 min that were higher than the minimal inhibitory concentration of the main equine respiratory pathogens. These results should stimulate further studies, especially in horses suffering from bronchial hyper‐reactivity. Aerosol delivery of antibiotics may well have a role in equine therapeutics.