BackgroundMultiple cytological patterns occur in bronchoalveolar lavage fluid (BALF) of horses with inflammatory airway disease (IAD). Only few data on BALF cytokine profiles are available for horses with IAD, and are limited to mRNA expression.Hypothesis/ObjectiveCytological profiles of IAD are associated with different BALF immunological pathways. To investigate BALF cytokine concentrations in a large number of horses with neutrophilic IAD.AnimalsOne hundred and thirty‐eight client‐owned Standardbred racehorses in active training.MethodsProspective observational study. BALF samples were obtained from left and right lungs. Interleukin (IL)‐4, interferon (IFN)‐γ, and tumor necrosis factor (TNF)‐α concentrations were determined by ELISA.ResultsFourteen horses had normal BALF cytological profiles and 56 exhibited evidence of bilateral neutrophilic IAD. Twenty‐four horses showed BALF with, respectively, IAD‐ and CTL consistent cytology and were excluded; as were 44 horses because of evidence of pulmonary hemorrhage. TNF‐α (56 ± 115 pg/mL; P = .034) and IFN‐γ concentrations (104 ± 247 pg/mL; P = .044) were significantly higher for IAD horses, compared with controls (respectively 19 ± 41 and 80 ± 116 pg/mL). Horses with ‘neutrophil’ subtype had significantly higher IFN‐γ concentrations (110 ± 154 pg/mL), than ‘neutrophil/metachromatic’ (56 ± 54 pg/mL; P = .028) and ‘neutrophil/metachromatic/eosinophil’ subtypes (44 ± 23 pg/mL; P = .012).Conclusions and Clinical ImportanceCytokine concentrations in BALF suggested that neutrophilic IAD is associated with activation of the innate immune system and a possible T‐helper (Th)‐1 polarized response. This study also suggested that immunological pathways vary according to cytological IAD subtypes.
The atlanto-occipital site (AO) is convenient for retrieving an adequate volume and quality of cerebrospinal fluid (CSF) in the diagnosis of neurological disease in horses. However, general anaesthesia is not always possible for horses displaying severe neurological signs, or for economical reasons. The objectives of the present work were to determine the feasibility and safety of ultrasound-guided CSF puncture at the AO site on the standing horse. Seven horses (six healthy and one mildly ataxic) were sedated with acepromazine (0.02 mg/kg bodyweight intravenously or 0.04 mg/kg bodyweight intramuscularly) and detomidine (0.01 mg/kg bodyweight intravenously), and placed in stocks or in a recovery stall with the head kept on a headstand. Puncture was performed by ultrasonographic guidance with a parasagittal technique, as previously described, using a 20 g, 3.5 inch spinal needle. In all horses, no adverse reaction was observed when crossing the dura mater and 20 ml of CSF was rapidly retrieved without any blood contamination. Ultrasound-guided CSF puncture can be performed easily at the AO site on a healthy standing horse. Regarding the potential risk of this procedure, safety measures and close observation are essential. Further studies on a larger amount of ataxic horses are also required before considering this technique as an alternative option for CSF puncture.
Mild to moderate equine asthma is prevalent in young racehorses, particularly early in their training period. Although the precise aetiopathogenesis remains undetermined, it is possible that the susceptibility of this population might partly reflect an exercise-associated immune derangement at the level of the airway. We performed a genome-wide basal gene expression scan on alveolar macrophages (AMs) isolated from Standardbred racehorses prior to and after commencement of competition race training with a view to identifying any exerciseassociated gene expression modulation consistent with functional alterations which might reflect training-associated immunological derangement. Microarray technology was used to analyse the basal gene expression profiles of bronchoalveolar fluid-derived AMs, harvested from six systemically healthy Standardbred racehorses prior to (T0) and following (T1) entry into training. Additionally, AM LPS-induced TNF-α and IL-10 release at T0 and T1 was assessed. Although the data revealed significant inter-horse heterogeneity in relation to the magnitude of individual gene expression at each time-point, within each horse, several inflammatory related genes (e.g. chemokine ligands, interferons and NFKB) declined in expression from T0 to T1. Entry into training did not significantly alter AM LPS-induced TNF-α or IL-10 release. The data support a direct effect of training on AM basal gene expression, particularly with respect to immune-related genes. The pattern of trainingassociated differential gene expression may indicate relative downregulation of inflammatory-related genes, consistent with an immunosuppressive effect of training and an increased susceptibility to opportunistic pathogens.
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