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.     

Increased adenosine concentration in bronchoalveolar lavage fluid of horses with lower airway inflammation

Several reports have suggested a role for adenosine in the pathogenesis of chronic airway conditions and this has led to new therapeutic strategies to limit airway inflammation. In this study, detectable levels of adenosine in bronchoalveolar lavage (BAL) samples from 11 horses with non-infectious lower-airway inflammation and 14 healthy controls are reported, with significantly higher values in horses with airway inflammation. Although these increased levels did not correlate with changes in neutrophil percentage in BAL, a positive association between adenosine levels and signs of lower airway inflammation (clinical score) was observed. These novel findings support the hypothesis that adenosine may contribute to bronchoconstriction and also act as a pro-inflammatory mediator in the bronchoalveolar milieu of horses with airway inflammation. Further investigation of this axis could lead to new approaches for the treatment of highly prevalent lower airway inflammatory conditions in the horse.     

Efficacy of oral prednisolone and dexamethasone in horses with recurrent airway obstruction in the presence of continuous antigen exposure

Reasons for performing study: Orally administered prednisolone and dexamethasone are used commonly in the treatment of recurrent airway obstruction (RAO) in horses. However, the efficacy of prednisolone in improving pulmonary function during continuous antigen exposure has not been evaluated critically and there is little evidence supporting the efficacy of low‐dose oral dexamethasone in the same conditions. Hypothesis: Oral prednisolone and dexamethasone improve pulmonary function in RAO under conditions of continuous antigen exposure, and dexamethasone is more effective than prednisolone at commonly used dosages. Methods: Using a randomised crossover design, prednisolone (2 mg/kg bwt) and dexamethasone (0.05 mg/kg bwt) were administered per os, s.i.d. for 7 days, to 7 horses during clinical exacerbation of the disease. Maximal difference in transpulmonary pressure (ΔP_L), lung resistance (R_L) and elastance (E_L) were measured before and after 3 and 7 days of treatment. Results: Prednisolone and dexamethasone improved pulmonary function significantly. However, the improvement was of greater magnitude after 3 and 7 days of treatment with dexamethasone compared to prednisolone. Also, after 7 days of treatment with dexamethasone, ΔP_L and R_L were not different from values obtained when horses were on pasture, while all 3 pulmonary function parameters remained different from pasture values after prednisolone treatment. Conclusions: Both corticosteroids improve pulmonary function, in spite of continuous antigen exposure. However, oral dexamethasone at 0.05 mg/kg bwt is more effective than prednisolone at 2 mg/kg bwt in the treatment of RAO. Potential relevance: Prednisolone was shown, for the first time, to our knowledge, to improve the pulmonary function of horses with RAO in the presence of continuous antigen exposure. This study also demonstrates the efficacy of low‐dose oral dexamethasone in reversing airway obstruction in these conditions.     

Plasma interleukin-6 response is predictive for severity and mortality in canine systemic inflammatory response syndrome and sepsis

BACKGROUND: Sepsis is still a major cause of death in both human and veterinary medicine. Early diagnosis is essential for appropriate treatment. Identification of patients at risk for developing sepsis is already possible in human medicine through the measurement of plasma interleukin-6 (IL-6) levels. In veterinary medicine, however, this has been investigated only in canine experimental models. OBJECTIVES: The purpose of this study was to measure IL-6 plasma levels in dogs with naturally occurring systemic inflammatory response syndrome (SIRS) and sepsis and to analyze the value of IL-6 as a predictive parameter for severity and mortality. METHODS: Included in the study were 79 dogs that had been admitted to the small animal clinics of Munich and Berlin from July 2004 to July 2005 and that satisfied the diagnostic criteria for SIRS and sepsis as defined using established parameters. Measurement of plasma IL-6 levels on days 0, 1, and 2 was performed by the use of a colorimetric bioassay based on IL-6-dependent cell growth. RESULTS: Septic foci were identified in 43 patients (septic group), and 36 patients were enrolled in the SIRS group. The frequency of positive blood cultures was 11%. The overall mortality rate was 48%. Higher plasma IL-6 levels on the day of admission were significantly correlated with a more severe degree of disease, increased mortality rate, and earlier fatality. CONCLUSIONS: Plasma IL-6 concentration is predictive of outcome in canine SIRS and sepsis and may be a valuable laboratory parameter for assessing critically ill dogs.     

Descriptive results from a longitudinal study of airway inflammation in British National Hunt racehorses

Reasons for performing study: Rates of airway inflammation in young racehorses decrease with time but it is not clear whether this is associated with increasing age or time exposed to the training environment. The structure of the British National Hunt (NH) population allowed closer examination of this relationship. Objectives: To compare rates of inflammatory airway disease diagnosed by tracheal sampling (trIAD), and its components, in NH racehorses by age and training history and with published rates in young racehorses. Methods: A prospective, longitudinal study was conducted on 5 NH yards over 2 years. Period sample prevalences of nasal discharge, tracheal mucus, airway neutrophilia and trIAD (defined by a combination of tracheal mucus and airway neutrophilia) were estimated and compared between horses with different ages and time in training. Results: Horses new to training had twice the odds of visible tracheal mucus as ex‐flat trained horses (OR 2.0; 95% CI: 1.4–2.8; P<0.001) but no significantly increased odds of airway neutrophilia (OR 1.3; 95% CI: 0.8–1.9; P = 0.3) and inconclusive evidence of increased odds of trIAD (OR 1.8; CI: 0.9–3.5; P = 0.08). However, a lower median time in training was significantly associated with the presence of visible mucus (P<0.001), increased mucus (P = 0.005) and trIAD (P = 0.03). No disease measure varied significantly with age. Conclusions: Tracheal mucus and trIAD, but not neutrophilia detected in tracheal wash samples, were less prevalent in horses that had been exposed to the training environment for longer, explaining previously reported associations with age. Potential relevance: Neutrophil proportion in tracheal wash samples is not as useful a clinical tool as measures of visible tracheal mucus for identifying horses requiring treatment or changes in management. The inclusion of tracheal wash neutrophils in the assessment of equine airways, or at least their relative weighting in definitions of trIAD, should be re‐evaluated.     

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.     

Exhaled breath condensate hydrogen peroxide and pH for the assessment of lower airway inflammation in the horse

Measurement of hydrogen peroxide (H₂O₂) concentration and pH in exhaled breath condensate (EBC) is useful for detection and monitoring of asthma in humans. In contrast, limited information on the use of these parameters for the investigation of lower airway inflammation (LAI) is available for horses. Aims of the current study were to investigate the intra- and inter-day variations of EBC H₂O₂ concentration and pH in horses and establish any relationship(s) with LAI. Both intra- and inter-day variability of EBC H₂O₂ concentration were large, while those of pH were small. No significant difference in the intra-day or inter-day H₂O₂ concentrations or pH measurements were found in control or LAI horses, except for inter-day H₂O₂ concentration in horses with LAI (p = 0.019). There was no significant difference in EBC pH or H₂O₂ concentration between control and LAI horses, however a trend for a reduced pH in horses with LAI was observed.     

Effect of ventriculectomy versus ventriculocordectomy on upper airway noise in draught horses with recurrent laryngeal neuropathy

Reasons for performing study: Little is known about the efficacy of bilateral ventriculectomy (VE) or bilateral ventriculocordectomy (VCE) in draught horses. Objectives: To compare the effect of VE and VCE on upper airway noise in draught horses with recurrent laryngeal neuropathy (RLN) by use of quantitative sound analysis techniques. Hypothesis: In competitive draught horses with grade 4 RLN, VE and VCE reduce upper airway noise during exercise, but VCE is more effective. Methods: Thirty competitive hitch or pulling draught horses with grade 4 RLN were evaluated for upper airway sound during exercise. Respiratory rate (RR), inspiratory (Ti) and expiratory time (Te), the ratio between Ti and Te (Ti/Te), inspiratory (Sli) and expiratory sound levels (Sle), the ratio between Sli and Sle (Sli/Sle), and peak sound intensity of the second formant (F2) were calculated. Eleven horses were treated with VE and 19 with VCE. After 90 days of voice and physical rest and 30 days of work, the horses returned for post operative upper airway sound evaluation and resting videoendoscopy. Results: VE significantly reduced Ti/Te, Sli, Sli/Sle and the sound intensity of F2. Respiratory rate, Ti, Te and Sle were unaffected by VE. VCE significantly reduced Ti/Te, Ti, Te, Sli, Sli/Sle and the sound intensity of F2, while RR and Sle were unaffected. The reduction in sound intensity of F2 following VCE was significantly greater than following VE. After VE and VCE, 7/11 (64%) and 15/18 (83%) owners, respectively, concluded that the surgery improved upper airway sound in their horses sufficiently for successful competition. Conclusions: VE and VCE significantly reduce upper airway noise and indices of airway obstruction in draught horses with RLN, but VCE is more effective than VE. The procedures have few post operative complications. Potential relevance: VCE is recommended as the preferred treatment for RLN in draught horses. Further studies are required to evaluate the longevity of the procedure's results.     

Hindlimb laminar inflammatory response is similar to that present in forelimbs after carbohydrate overload in horses

Reasons for performing study: A significant proinflammatory response is known to occur in the forelimb lamina after carbohydrate administration. As the hindlimbs are often less affected by laminitis compared with the forelimbs, we assessed hindlimb inflammatory response in the early stages of carbohydrate‐induced laminitis to determine whether differences in the response existed. Objective: To determine whether a similar proinflammatory response occurs in the hindlimb laminae to that previously reported for the forelimb. Methods: Archived laminar samples from 12 horses administered 17.6 g of starch (85% corn starch, 15% wood flour)/kg bwt via nasogastric tube that were anaesthetised either after developing a temperature >38.9°C (DEV; n = 6) or at the onset of Obel grade 1 lameness (OG1; n = 6) were used in addition to 6 control horses (CON) that were anaesthetised 24 h after administration of water. Real‐time quantitative polymerase chain reaction for selected proinflammatory mediators and MAC387 immunohistochemistry were performed. The data were analysed nonparametrically to compare groups. Results: Increases in laminar MAC387‐positive leucocytes and laminar messenger ribonucleic acid (mRNA) concentrations (P<0.05) for interleukin‐1β, interleukin‐6, cyclo‐oxygenase‐2, chemokine (C‐X‐C motif)ligand (CXCL)1 and CXCL8 were present in both fore‐ and hindlimb laminae from horses with OG1 lameness. Both CXCL1 and CXCL8 were also increased in forelimb and hindlimb laminae in the DEV horses. Conclusions: Administration of carbohydrate resulted in a similar inflammatory response in the hindlimb laminae to that previously reported for the forelimb laminae. These findings suggest that other factors, such as weightbearing, may play an important role in the development of laminitis after a systemic inflammatory condition develops. Potential relevance: Evidence of inflammation in the hindlimb laminae suggests that the hindfeet should be addressed in the septic horse at risk for laminitis; however, laminitis is often less severe in the hindlimbs due to other factors, such as weightbearing and hoof angle.     

Comparison of post-operative inflammatory response in horses undergoing elective castration treated preoperatively with ceftiofur crystalline free acid or procaine penicillin G

Castration is among the most common surgical procedures performed in the horse (Equus Caballus) and a variety of post-operative complications can occur. This study aims to determine if a single dose of long-acting ceftiofur crystalline free acid (CCFA) used as a preoperative antimicrobial in equine field castrations offers any reduction in post-operative inflammatory markers when compared to procaine penicillin G (PPG). Sixty-five horses aged 8 months to 2 years were randomly assigned to the CCFA (n = 33) or PPG (n = 32) treatment groups. Horses were castrated under general anaesthesia using a closed castration technique with removal of the median raphe. Quantitative and qualitative inflammatory markers were measured and short-term complications were recorded post-operatively on Days 3, 8 and 14. No clinically significant difference in any post-operative inflammatory markers between the CCFA and PPG group was detected. In the CCFA group, 48% of horses experienced short-term post-operative complications compared to 31% in the PPG group. Regardless of the preoperative treatment, castration induced significant elevation in serum amyloid A (P<0.0001), preputial oedema (P<0.0001) and scrotal oedema (P<0.0001) at Day 3. These values returned to baseline levels by Day 8. Horses with grade 3 or above preputial oedema had elevated serum amyloid A values (P<0.001). The data from this study indicate CCFA used as a preoperative antibiotic for routine castration offers no advantages over PPG. The difference in complication rate between groups is likely of minimal clinical importance, as all complications were mild and self-limiting.     

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.     

Laser vocal cordectomy fails to effectively reduce respiratory noise in horses with laryngeal hemiplegia

OBJECTIVE: To report the effect of unilateral laser vocal cordectomy on respiratory noise and airway function in horses with experimentally induced laryngeal hemiplegia (LH). STUDY DESIGN: Experimental study. ANIMALS: Six Standardbred horses without upper airway abnormalities at rest or during high-speed treadmill exercise. METHODS: Respiratory sounds and inspiratory trans-upper airway pressure (P(Ui)) were measured before (baseline) and 14 days after induction of LH by left recurrent laryngeal neurectomy, and again 30, 60, 90, and 120 days after endoscopically assisted laser cordectomy of the left vocal cord. Data were collected with the horses exercising on a treadmill at a speed producing maximum heart rate (HR(max)). RESULTS: In horses exercising at HR(max), induction of LH caused a significant increase in P(Ui), sound level (SL), and the sound intensity of formant 2 (F(2)) and 3 (F(3)). The sound intensity of formant 1 (F(1)) was unaffected by induction of LH. Laser vocal cordectomy had no effect on SL, or on the sound intensity of F(1) and F(3). At 30, 60, 90, and 120 days after surgery, P(Ui) and the sound intensity of F(2) were significantly reduced, but these variables remained significantly different from baseline values. CONCLUSIONS: Unilateral laser vocal cordectomy did not effectively improve upper airway noise in horses with LH. The procedure decreased upper airway obstruction to the same degree as bilateral ventriculocordectomy. CLINICAL RELEVANCE: Currently, laser vocal cordectomy cannot be recommended for the treatment of upper airway noise in horses with LH.