The Use of Bi-Nasal Prongs for Delivery of Non-Invasive Ventilation to Foals.

Non-invasive ventilation (NIV) is a method of providing respiratory support without the need for airway intubation. The current study was undertaken to assess tolerance to bi-nasal prongs and NIV in healthy, standing, lightly sedated foals. Bi-nasal prongs were well tolerated by foals, remaining in place for the allocated five minutes in four of six unsedated foals and, subsequently, in five of six lightly sedated foals. All foals tolerated NIV through bi-nasal prongs, although increasing airway pressures were associated with increases in inspiratory volume, duration of inspiration and air leakage in most foals. These changes preceded discontinuation/intolerance of NIV on the basis of behaviour changes consistent with discomfort. Increased circuit leakage was associated with reduced return of expired air to the ventilator and increasing disparity between inspiratory and expiratory times and tidal volumes. The study results suggest that bi-nasal prongs might be suitable for NIV but that design or fitting requires further optimization and that behaviour and ventilator variables should be monitored to assess patient tolerance of the procedure.

Wound healing of experimental equine skin wounds and concurrent microbiota in wound dressings following topical propylene glycol gel treatment.

Introduction: Topical wound treatments rely on carrier formulations with little to no biological impact. The potential for a common vehicle, a propylene glycol (PG) gel, to affect wound healing measures including microbiota is not known. Microbiome characterization, based on next generation sequencing methods is typically performed on tissue or directly obtained wound fluid samples. The utility for primary wound dressings to characterize equine wound microbiota in the context of topical treatments is currently unknown. This investigation reports the topical effect of an 80% PG based gel on wound healing and microbiota in wound dressings. Methods: Experiments were performed in six mature horses utilizing a surgical, distal limb wound model, histology of sequential wound biopsies, photographic wound measurements and microbiota profiling via 16s rRNA sequencing of wound dressing samples. Experimental wounds were surveyed for 42 days and either treated (Day 7, 14, 21 and 28; at 0.03 ml/cm2) or unexposed to the PG gel. Wound surface area, relative and absolute microbial abundances, diversity indices and histologic parameters were analyzed in the context of the experimental group (treatment; control) using qualitative or quantitative methods depending on data characteristics. Results: Compared to controls, treatment slowed the wound healing rate (17.17 ± 4.27 vs. 18.56 ± 6.3 mm2/day), delayed the temporal decline of polymorphonucleated cells in wound beds and operational taxonomic units (OTU) in wound dressings and lowered alpha-diversity indices for microbiota in primary wound dressing. Relative abundances of OTUs were in line with those previously reported for equine wounds. Clinical outcomes 42 days post wounding were considered similar irrespective of PG gel exposure. Discussion: Results highlight the potential for vehicle exposure to alter relevant wound outcome measures, imposing the need for stringent experimental control measures. Primary wound dressings may represent an alternate sample source for characterization of the wound microbiome alleviating the need for additional interventions. Further studies are warranted to contrast the microbiome in wound dressings against that present on wound surfaces to conclude on the validity of this approach.

Visualizing neutrophil extracellular traps in septic equine synovial and peritoneal fluid samples using immunofluorescence microscopy.

Septic synovitis and peritonitis are routinely diagnosed in horses based on clinical examination findings and laboratory assessment of synoviocentesis and abdominocentesis samples, respectively. Diagnosis is difficult in some cases because of an overlap in laboratory results for septic and non-septic inflammation. Neutrophil extracellular trap (NET) formation is part of the innate immune response against pathogens. Identifying and quantifying NETs, which have not been explored in clinical samples from horses with septic synovitis and peritonitis, to our knowledge, may be helpful in detecting infectious processes. Our main objective was to determine whether NETs could be visualized in septic equine synovial and peritoneal fluid cytology samples using immunofluorescence with antibodies against citrullinated histone H3 (Cit-H3) and myeloperoxidase (MPO). We analyzed 9 synovial and 4 peritoneal fluid samples. NET percentages were quantified using a simple counting technique, which is suitable for high-quality, well-preserved, and stained cytospin smears. NETs were evident in all septic samples and were absent in a non-septic sample; NETs were better visualized with Cit-H3 than with MPO immunolabeling. Overall, we believe that there is the potential for NETs and associated markers to be used to investigate and understand septic inflammation in horses.

Effects of 2 modes of positive pressure ventilation on respiratory mechanics and gas exchange in foals.

BACKGROUND: Continuous positive airway pressure (CPAP) and pressure support ventilation (PSV) can improve respiratory mechanics and gas exchange, but different airway pressures have not been compared in foals. HYPOTHESIS/OBJECTIVES: Assess the effect of different airway pressures during CPAP and PSV have on respiratory function in healthy foals with pharmacologically induced respiratory insufficiency. We hypothesized that increased airway pressures would improve respiratory mechanics and increased positive end-expiratory pressure (PEEP) would be associated with hypercapnia. ANIMALS: Six healthy foals from a university teaching herd. METHODS: A prospective, 2-phase, 2-treatment, randomized cross-over study design was used to evaluate sequential interventions in sedated foals using 2 protocols (CPAP and PSV). Outcome measures included arterial blood gases, spirometry, volumetric capnography, lung volume and aeration assessed using computed tomography (CT). RESULTS: Sedation and dorsal recumbency were associated with significant reductions in arterial oxygen pressure (PaO2 ), respiratory rate, and tidal volume. Continuous positive airway pressure was associated with improved PaO2 , without concurrent hypercapnia. Volumetric capnography identified improved ventilation:perfusion (V/Q) matching and increased carbon dioxide elimination during ventilation, and spirometry identified decreased respiratory rate and increased tidal volume. Peak inspiratory pressure was moderately associated with PaO2 and lung volume. Improved pulmonary aeration was evident in CT images, and lung volume was increased, particularly during CPAP. CONCLUSIONS AND CLINICAL IMPORTANCE: Both CPAP and PSV improved lung mechanics and gas exchange in healthy foals with induced respiratory insufficiency.

Effects of Transport and Feeding Strategies Before Transportation on Redox Homeostasis and Gastric Ulceration in Horses.

Transportation may lead to oxidative stress (OS) and gastric ulceration in horses, and optimal feed management before, or during, transportation is unclear. This study aimed to evaluate the effects of transportation after three different feeding strategies on OS and to explore possible associations between OS and equine gastric ulcer syndrome (EGUS). Twenty-six mares were transported by truck for 12 hours without food or water. Horses were randomly divided into 3 groups; (1) fed 1 hour before departure (BD), (2) fed 6 hours BD, (3) fed 12 hours BD. Clinical examinations and blood collections were performed at approximately 4 hours BD (T0), at unloading (T1), 8 hours (T2) and 60 hours (T3) after unloading. Gastroscopy was conducted prior to departure, and at T1 and T3. Although OS parameters remained in the normal range, transportation was associated with increased reactive oxygen metabolites (ROMS) at unloading (P=0.004), with differences between horses fed 1 hour and 12 hours BD (P < .05). The level of total antioxidant (PTAS) was affected by both transportation and feeding strategy (P = 0.019), with horses fed 1 hour BD demonstrating greater PTAS at T = 0, and a different response in comparison with the other groups and the literature. Nine horses demonstrated clinically significant ulceration of the squamous mucosa at T1 but, although weak correlations were evident between OS parameters and ulcer scores, univariate logistic regression showed no associations. This study suggests that feed management prior to a long journey (12 hours) may affect oxidative balance. Further studies are needed to understand the nexus between feed management before and during transport, transport-related OS and EGUS.

Impact of sedation, body position change and continuous positive airway pressure on distribution of ventilation in healthy foals.

Background: This study aimed to compare the distribution of ventilation measured by electrical impedance tomography (EIT), in foals under varying clinical conditions of sedation, postural changes, and continuous positive airway pressure (CPAP). To support the interpretation of EIT variables, specific spirometry data and F-shunt calculation were also assessed. Materials and methods: Six healthy Thoroughbred foals were recruited for this sequential experimental study. EIT and spirometry data was recorded: (1) before and after diazepam-sedation, (2) after moving from standing to right lateral recumbency, (3) in dorsal recumbency during no CPAP (CPAP0) and increasing levels of CPAP of 4, 7, and 10 cmH2O (CPAP4, 7, 10, respectively). Ventral to dorsal (COVVD) and right to left (COVRL) center of ventilation, silent spaces, tidal impedance variation, regional ventilation distribution variables and right to left lung ventilation ratio (R:L) were extracted. Minute ventilation was calculated from tidal volume (VT) and respiratory rate. F-Shunt was calculated from results of arterial blood gas analysis. Statistical analysis was performed using linear mixed effects models (significance determined at p < 0.05). Results: (1) Respiratory rate was lower after sedation (p = 0.0004). (2) In right lateral recumbency (compared to standing), the COVVD (p = 0.0012), COVRL (p = 0.0057), left centro-dorsal (p = 0.0071) and dorsal (p < 0.0001) regional ventilation were higher, while the right ventral (p = 0.0016) and dorsal (p = 0.0145) regional ventilation, and R:L (p = 0.0017) were lower. (3) Data of two foals for CPAP10 was excluded from statistical analysis due to prolonged apnea. Stepwise increase of CPAP led to increases of COVVD (p = 0.0028) and VT (p = 0.0011). A reduction of respiratory rate was detected with increasing CPAP levels (p < 0.0001). Conclusions: (1) In healthy foals, diazepam administration did not alter distribution of ventilation or minute ventilation, (2) lateral recumbency results in collapse of dependent areas of the lung, and (3) the use of CPAP in dorsal recumbency at increasing pressures improves ventilation in dependent regions, suggesting improvement of ventilation-perfusion mismatch.

Bi-Level Positive Airway Pressure for Non-invasive Respiratory Support of Foals.

Respiratory insufficiency and pulmonary health are important considerations in equine neonatal care. As the majority of foals are bred for athletic pursuits, strategies for respiratory support of compromised foals are of particular importance. The administration of supplementary oxygen is readily implemented in equine practice settings, but does not address respiratory insufficiency due to inadequate ventilation and is no longer considered optimal care for hypoxia in critical care settings. Non-invasive ventilatory strategies including continuous or bi-level positive airway pressure are effective in human and veterinary studies, and may offer improved respiratory support in equine clinical practice. The current study was conducted to investigate the use of a commercial bi-level positive airway pressure (BiPAP) ventilator, designed for home care of people with obstructive respiratory conditions, for respiratory support of healthy foals with pharmacologically induced respiratory insufficiency. A two sequence (administration of supplementary oxygen with, or without, BiPAP), two phase, cross-over experimental design was used in a prospective study with six foals. Gas exchange and mechanics of breathing (increased tidal volume, decreased respiratory rate and increased peak inspiratory flow) were improved during BiPAP relative to administration of supplementary oxygen alone or prior studies using continuous positive airway pressure, but modest hypercapnia was observed. Clinical observations, pulse oximetry and monitoring of expired carbon dioxide was of limited benefit in identification of foals responding inappropriately to BiPAP, and improved methods to assess and monitor respiratory function are required in foals.

Decreased Virus-Neutralizing Antibodies Against Equine Herpesvirus type 1 In Nasal Secretions of Horses After 12-hour Transportation.

This study evaluated the effects of 12-hour transportation on immune responses to equine herpesvirus type 1 (EHV-1) and type 4 (EHV-4). Possible replication of EHV-1 and EHV-4 was monitored by real-time PCR of nasal swabs and peripheral blood mononuclear cells (PBMCs), and changes in systemic and mucosal antibodies were investigated. Six healthy Thoroughbreds with transport experience were transported in commercial trucks, repeating the same three-hour route four times. Blood samples for cortisol measurement were taken before departure and every three hours. Nasal swabs, PBMCs, nasal wash and serum samples were collected before departure, at unloading, two and six days after arrival. Cortisol concentration increased significantly after three and six hours of transport (P < 0.05), confirming acute transport stress. However, no evidence of viral replication or lytic infection was observed, and serum virus neutralization (VN) titers for EHV-1 and EHV-4 were unchanged, except for one horse that showed a four-fold decrease in titer against EHV-1 after transportation. Urea and total IgA concentration in nasal washes increased significantly after transportation (P < 0.05), while total IgA/protein ratio was unchanged. A transient, ≥4-fold decrease in VN titers for EHV-1 in nasal wash concentrates was observed in four out of six horses after transportation (geometric mean titer declined from 202 to 57, P < 0.05), suggesting suppression of VN capacity in the nasal mucosa may contribute to susceptibility to EHV-1 after transportation. VN antibodies against EHV-4 in nasal secretion were not detected at any timepoint.

Pharmacokinetic and pharmacodynamic effects of 2 registered omeprazole preparations and varying dose rates in horses.

BACKGROUND: Omeprazole preparations vary in bioavailability in horses. HYPOTHESIS/OBJECTIVES: To characterize the pharmacokinetics and pharmacodynamics of an existing enteric-coated oral omeprazole paste (REF) and a novel, in-feed, enteric-coated dry granule preparation (NOV). ANIMALS: Twelve Standardbred/Thoroughbred mares free from clinical disease. METHODS: A prospective, blinded randomized interventional study was trial, conducted in 3 parts: (a) bioavailability study, (b) dose titration study, and (c) comparative clinical pharmacodynamic study, each using a blocked crossover design. RESULTS: Consistent with the larger dose administered, Cmax (median, 1032 ng/mL; range, 576-1766) and AUC0-24 (median, 63.9 µg/mL*min; range, 42.4-152.4) were greater after single oral administration of NOV than REF (282.7 ng/mL; range, 94.8-390.2, and 319 23.8 µg/mL*min; range, 8.2-42.3, respectively; both P = .004). No differences were observed between products for absolute oral bioavailability (NOV 55% range, 15-88; REF 17% range, 10-77; P = .25). Treatment with both preparations was associated with reduced gastric squamous ulcer scores and increased pH of gastric fluid. Bioequivalence was demonstrated for pharmacodynamic measures with the exception of % time pH <4, despite differences in dose rate and subsequent plasma omeprazole concentrations. CONCLUSIONS AND CLINICAL IMPORTANCE: The findings of this study indicate that the NOV product would be a suitable alternative to the reference product, and confirm that plasma concentrations of omeprazole and omeprazole dose do not predict drug pharmacodynamics in horses.

Interobserver and intraobserver reliability for 2 grading systems for gastric ulcer syndrome in horses.

BACKGROUND: Grading of equine gastric ulcer syndrome (EGUS) is undertaken in clinical and research settings, but the reliability of EGUS grading systems is poorly understood. HYPOTHESIS/OBJECTIVES: Investigate interobserver and intraobserver reliability of an established ordinal grading system and a novel visual analog scale (VAS), and assess the influence of observer experience. ANIMALS: Sixty deidentified gastroscopy videos. METHODS: Six observers (3 specialists and 3 residents) graded videos using the EGUS Council (EGUC) system and VAS. Observers graded the videos three 3 for each system, using a cross-over design with at least 1 week between each phase. The order of videos was randomized for each phase. METHODS: Interobserver and intraobserver reliability were estimated using Gwet's agreement coefficient with ordinal weights applied (AC2) for the EGUC system and the intraclass correlation coefficient (ICC) for the VAS. RESULTS: Using the EGUC system, interobserver reliability was substantial for squamous (AC2 = 0.69; 95% confidence interval [CI], 0.57-0.80) and glandular mucosa (AC2 = 0.72; 95% CI, 0.70-0.75), and intraobserver reliability was substantial for squamous (AC2 = 0.80; 95% CI, 0.71-0.90) and glandular mucosa (AC2 = 0.80; 95% CI, 0.74-0.86). Interobserver reliability using the VAS was moderate for squamous (ICC = 0.64; 95% CI, 0.31-0.96) and poor for glandular mucosa (ICC = 0.35; 95% CI, 0.06-0.64), and intraobserver reliability was moderate for squamous (ICC = 0.74; 95% CI, 0.62-0.86) and glandular mucosa (ICC = 0.56; 95% CI, 0.39-0.72). CONCLUSIONS AND CLINICAL IMPORTANCE: The EGUC system had acceptable intraobserver and interobserver reliability and performed well regardless of observer experience. Familiarity and observer experience improved reliability of the VAS.

Intragastric pH of foals admitted to the intensive care unit.

BACKGROUND: Intragastric pH profiles of neonatal foals admitted to the intensive care unit (ICU) remain poorly characterized. HYPOTHESIS/OBJECTIVES: To determine intragastric pH profiles and clinical parameters associated with intragastric pH in foals admitted to the ICU. ANIMALS: Forty-two neonatal foals admitted to the ICU and requiring placement of an indwelling nasogastric tube for nutritional management were included. METHODS: Intragastric pH was measured for 24 hours from the time of admission. Mean pH, % time pH <4, and % time pH >4 were determined for each foal. History, clinical findings, and clinicopathological data recorded at the time of presentation were collected. RESULTS: The mean pH of included foals was 5.5 ± 1.8. The median % time pH <4 was 6.3% (range: 0-99). A history of placentitis was associated with greater mean pH (median 5.3 (range: 0.9-7.8) versus median 7.2 (5.9-11.3); P = .002) and less % time pH <4 (median 13 (0-99.6) versus median 0.1 (0-7.2); P = .01). Foals with diarrhea had a greater % time pH <4 (median 4.6% (0-99) versus median 28.8% (1.4-57.48); P = .02). Foals with a pH >4 for >50% recording time had a lower PaO2 (mean difference 25.0 mm Hg; 95% confidence interval [CI], 14.4-35.6; P = .03) and higher PaCO2 (mean difference 14.9 mm Hg; 95% CI, 4.7-25.2; P = .02). Surviving foals had a lower mean median hourly pH (P = .02). CONCLUSIONS AND CLINICAL IMPORTANCE: Intragastric pH profiles were unpredictable and mostly >4 for >80% of the recording time. This study does not support the indiscriminate administration of acid suppressive treatment.

Effects of transportation on gastric pH and gastric ulceration in mares.

BACKGROUND: Transportation has been suggested as a risk factor for gastric ulceration in horses, but limited evidence supports this assumption. ANIMALS: Twenty-six Standardbred, Thoroughbred, and Warmblood mares from a university teaching herd. METHODS: Twelve mares were confined for 12 hours, overnight, in reproductive stocks with indwelling nasogastric tubes (NGTs) to assess pH of gastric fluid (GF). Gastric ulceration was assessed endoscopically before and after confinement. Subsequently, 26 horses were transported for 12 hours, overnight, in 2 consignments. During transportation, GF was aspirated from indwelling NGT placed in the same 12 mares used in the confinement study, and gastric ulceration was assessed endoscopically before and after transportation in all horses. RESULTS: The median pH of GF in confined horses was 1.70-2.49 at each sampling point, and there was no apparent effect on gastric squamous ulcer scores. The median pH of GF from the same 12 horses at corresponding sampling times during transportation was 6.82-7.22. Transportation was associated with increased gastric squamous ulcer scores, particularly in horses fasted for gastroscopy and NGT placement immediately before departure. Gastric emptying appeared delayed after transportation in horses fed before departure. CONCLUSIONS AND CLINICAL IMPORTANCE: Transportation is associated with increased gastric squamous ulceration and with increased pH of GF. These findings may be a consequence of impaired gastric emptying and reflux of alkaline small intestinal content, with factors such as duodenal bile salts and short-chain fatty acids mediating mucosal injury.

Effects of Transport Conditions on Behavioural and Physiological Responses of Horses.

The regulations for minimal space and direction of travel for land transport in horses vary worldwide and there is currently no definitive guidance to promote equine health and welfare. This study evaluated the effects of bay size and direction of travel (forwards/backwards) in horses by comparing the behavioural, physiological, laboratory and gastroscopy parameters between transported and confined horses. A total of twenty-six mares took part in the study; 12 horses were confined for 12 h, and all mares underwent 12 hours' transportation, travelling in single (n = 18) or wide bays (n = 8), and forward (n = 10) or rear (n = 16) facing. Behaviour was recorded during confinement/transportation and analysed using a behaviour sampling ethogram. Clinical examination, blood samples and gastroscopy were conducted before and after confinement/transportation. The frequency of behaviours relating to stress and balance increased during transport, and horses transported in a rear-facing position and in a wider bay size showed fewer balance-related behaviours. Balance behaviours, particularly loss of balance, were positively associated with the severity of gastric ulceration after transportation and elevated muscle enzymes, while increased stress behaviours correlated with decreased gastrointestinal sounds. Heart rate and rectal temperature after transportation were positively associated with balance and stress behaviours, and with squamous gastric ulcer scores. Transportation was associated with expected increases in cortisol and muscle enzymes, but positioning and space allowance had minimal effects on these analytes. Findings suggest that transportation in a rear-facing position and in wider bays might reduce the impact of transport on horse health and welfare, and monitoring behaviour in transit and physiological measurements after transportation should be recommended. Behavioural and physiological parameters were more sensitive than haematological, biochemical or endocrine analytes to identify horses suffering from transport stress.

A questionnaire study of parasite control in Thoroughbred and Standardbred horses in Australia.

BACKGROUND: Information regarding parasite control practices currently used on Thoroughbred and Standardbred studs in Australia is lacking. Anthelmintic resistance (AR) is a global problem which has implications for equine health and welfare. OBJECTIVES: To identify parasite control practices currently used on horse studs in Australia and investigate the frequency of use of management factors that have been associated with the likelihood of promoting or delaying AR. STUDY DESIGN: Questionnaire study of equine parasite control on Thoroughbred and Standardbred studs in Australia. METHODS: An online questionnaire was emailed to 300 studs in Australia. Information obtained included property details, grazing management, anthelmintic use, non-chemotherapeutic parasite control practices, use of faecal egg counts (FECs) and perceptions of AR. RESULTS: Seventy-five completed questionnaires were received (25% response rate). Macrocyclic lactones (MLs) were the most commonly administered anthelmintics in mares and foals and less than 5% of respondents used targeted treatment regimens. The implementation of pasture hygiene practices was variable. The majority of respondents (97%) considered AR to be important; however, few respondents were aware of the use of FEC reduction tests for monitoring of drug efficacy. MAIN LIMITATIONS: The potential for nonresponse bias was the main limitation of this study. CONCLUSIONS: Parasite control strategies on Australian stud farms remain over-reliant on anthelmintic use. The frequent use of MLs is of concern for the increased selection pressure for AR. There is a lack of awareness of the importance of non-chemotherapeutic strategies in integrated approaches to parasite control aimed at delaying the development of AR. This study highlights the need for greater veterinary involvement in the implementation of more sustainable parasite control practices with greater emphasis placed on surveillance through FEC testing.

Effects of continuous positive airway pressure on respiratory function in sedated foals.

OBJECTIVE: To characterize the effects of continuous positive airway pressure (CPAP) delivered by a commercial human CPAP device on respiratory function in foals with pharmacologically induced respiratory suppression. DESIGN: Prospective randomized, cross-over study comparing CPAP with spontaneous respiration and oxygen insufflation. SETTING: University veterinary teaching hospital. ANIMALS: Twelve foals born in consecutive seasons from a university teaching herd. INTERVENTIONS: Foals were randomized to receive 10 minutes of respiratory support by mask oxygen supplementation or CPAP as a first treatment after induction of respiratory depression by intravenous administration of xylazine and fentanyl. Each foal received the alternate treatment after 10 minutes of breathing ambient air, and the procedure was repeated after 48 hours with treatment order reversed. MEASUREMENTS AND MAIN RESULTS: The administration of xylazine and fentanyl by bolus or continuous infusion reliably induced reversible respiratory suppression and recumbency. CPAP was associated with comparable increase in PaO2 relative to mask oxygen supplementation, but with lower respiratory rate, increased oxygen extraction and increased carbon dioxide elimination. Mild increase in PaCO2 was observed during CPAP and O2 supplementation. Expiratory time increased and peak expiratory flow decreased during CPAP. CONCLUSIONS: Findings of the study suggest that CPAP might represent a method for improved respiratory support compared to O2 insufflation due to increased respiratory efficiency. Care must be taken in extrapolation of these findings from foals with pharmacologically induced respiratory compromise to foals with clinical respiratory disease, and further investigation is required to better characterize the cause and impact of marginal hypercapnia observed in these studies.

Equine Transport and Changes in Equid Herpesvirus' Status.

The risk of respiratory disease in the transported horse can increase as a consequence of immunosuppression and stress associated primarily with opportunistic bacterial proliferation and viral reactivation. This study examines the ecology of equid herpesviruses (EHV) in these horses, exploring reactivation and changes in infection and shedding associated with transport, and any potential contributions to transport-related respiratory disease. Twelve horses were subjected to an 8-h road-transport event. Antibodies to EHV-1 and EHV-4 were detected by ELISA in serum collected prior to, immediately after and 2 weeks post transport. Respiratory tract endoscopy and tracheal washes were collected prior to and 5 days after transportation. Nasal swabs collected prior to, immediately after, 1 and 5 days following transport were screened for EHV-1,-2,-4,-5 using qPCR. Six horses had persistent neutrophilic airway infiltrates post transportation, indicative of subclinical respiratory disease. No horses were qPCR positive for either of the alphaherpesviruses (i.e., EHV-1/-4) nor did any seroconvert to either virus. Four out of nine horses positive for either EHV-2 or EHV-5 on qPCR prior to transport developed neutrophilic airway inflammation. Five horses showed increasingly positive readings on qPCR (i.e., reduced Cq) for EHV-2 after transportation and seven out of eleven horses positive for EHV-2 after transport shared strains of high sequence similarity with other horses in the study. One EHV-2 virus detected in one horse after transport was genetically different which may be due to reactivation. The clinical significance of EHV-2 and EHV-5 remains in question. However these results indicate that transportation may lead to increased shedding, transmission and reactivation of EHV-2 and EHV-5 but not EHV-1/-4. Unlike previous work focusing on the role of alphaherpesviruses, this research suggests that investigation of the gammaherpesviruses (i.e., EHV-2/-5) in transport-related disease should not be dismissed, particularly given that these viruses can encode suppressive immunomodulators that may affect host health.

Behaviour during transportation predicts stress response and lower airway contamination in horses.

This study aimed to document the effects of an eight hour journey on behavioural, clinical, haematological, environmental and respiratory parameters, and to identify possible associations between factors. Twelve horses underwent clinical examination, respiratory endoscopy with tracheal wash (TW) aspiration, and collection of venous and arterial blood before (BJ) and after the journey (AJ). TW were submitted for conventional quantitative bacteriological evaluation and genetic microbiome analyses. Behaviour was assessed in stables prior to transportation and throughout the journey. Transportation caused mild, but significant, effects on fluid and electrolyte balance and an acute phase response, characterized by neutrophilia, hyperfibrinogenaemia and hyperglobulinaemia. The proportion of neutrophils in TW, tracheal mucus and TW bacterial concentration was increased AJ, with preferential replication of Pasteurellaceae. Horse behaviour en route predicted clinical and respiratory outcomes. The frequency of stress related behaviours was greatest in the first hour of the journey, and balance-related behaviours were most common in the final hour of the journey. Horses which lowered their heads less frequently en route and showed more stress-related behaviours had higher physiological stress (serum cortisol and heart rate on arrival), increased tracheal mucus and inflammation scores, and higher TW bacterial concentration AJ (P<0.05). Six horses with abnormal lung auscultation AJ proved to have had higher tracheal inflammation scores at preloading (P = 0.017), an overall higher concentration of bacteria in their TW (P = 0.013), and an increased percentage of neutrophils in TW at five days AJ (P = 0.003) in comparison to the other horses. While transport-related health problems are multifactorial, clinical examination, including auscultation and endoscopic inspection of the lower respiratory tract before and after journey, and behavioural observation en route may identify animals at increased risk of transport associated respiratory disease.

Salmeterol undergoes enantioselective bronchopulmonary distribution with receptor localisation a likely determinant of duration of action.

BACKGROUND: Salmeterol (a long acting beta2-agonist) is a chiral molecule. (RR)-salmeterol is responsible for pharmacological effect, but basic knowledge of enantioselective pulmonary pharmacodynamics and pharmacokinetics of salmeterol remains unknown. There are safety concerns with (S)-enantiomers of beta2-agonists, with suggestions that these enantiomers may increase bronchial hyperresponsivneness in asthma patients. METHODOLOGY: Horses (n = 12) received racemic (rac-) salmeterol 250 µg via inhalation. Enantioselective UPLC-MS/MS was used to determine (R)- and (S)-salmeterol concentrations in pulmonary epithelial lining fluid (PELF) sampled 2, 5, 10 and 15 min after administration, in central lung (endoscopic bronchial biopsy) and peripheral lung (percutaneous pulmonary biopsy) tissues (at 20 and 25 min respectively), and in plasma samples. RESULTS: Physiologically relevant tissue concentrations were found for both enantiomers, with median levels greater in central than peripheral lung (equivalent to 32 and 5 mM (R)-salmeterol for central and peripheral lung respectively). Levels in PELF decreased around 50% over 15 min and enantioselective distribution was observed in the central lung with levels of (R)-salmeterol around 30% higher than (S)-salmeterol. CONCLUSION: Salmeterol distribution is enantioselective in the central lung. This suggests duration of action is more likely associated with specific B2ADR localisation effects rather than non-specific physiochemical factors which would not be enantioselective.

Long-Acting ß2-Agonists in Asthma: Enantioselective Safety Studies are Needed.

Long-acting ß2-agonists (LABAs) such as formoterol and salmeterol are used for prolonged bronchodilatation in asthma, usually in combination with inhaled corticosteroids (ICSs). Unexplained paradoxical asthma exacerbations and deaths have been associated with LABAs, particularly when used without ICS. LABAs clearly demonstrate effective bronchodilatation and steroid-sparing activity, but long-term treatment can lead to tolerance of their bronchodilator effects. There are also concerns with regard to the effects of LABAs on bronchial hyperresponsiveness (BHR), where long-term use is associated with increased BHR and loss of bronchoprotection. A complicating factor is that formoterol and salmeterol are both chiral compounds, usually administered as 50:50 racemic (rac-) mixtures of two enantiomers. The chiral nature of these compounds has been largely forgotten in the debate regarding LABA safety and effects on BHR, particularly that (S)-enantiomers of ß2-agonists may be deleterious to asthma control. LABAs display enantioselective pharmacokinetics and pharmacodynamics. Biological plausibility of the deleterious effects of ß2-agonists (S)-enantiomers is provided by in vitro and in vivo studies from the short-acting ß2-agonist (SABA) salbutamol. Supportive clinical findings include the fact that patients in emergency departments who demonstrate a blunted response to salbutamol are more likely to benefit from (R)-salbutamol than rac-salbutamol, and resistance to salbutamol appears to be a contributory mechanism in rapid asthma deaths. More effort should therefore be applied to investigating potential enantiospecific effects of LABAs on safety, specifically bronchoprotection. Safety studies directly assessing the effects of LABA (S)-enantiomers on BHR are long overdue.

Immunological, clinical, haematological and oxidative responses to long distance transportation in horses.

Horses are transported frequently and often over long distances. Transportation may represent a physiological stressor with consequential health and welfare implications. This study reports the effects of a long distance journey on immunological, clinical, haematological, inflammatory and oxidative parameters in an Experimental Group (EG) of ten horses, comparing them with six horses of similar age and breed used as a non-transported Control Group (CG). Clinical examination and blood sampling were performed twice on all horses: immediately after unloading for the EG, and at rest on the same day for the CG (day 1); at rest on the same day one week later for both groups (day 7). On day 1 EG horses showed increased heart and respiratory rates (P<0.01), rectal temperature (P<0.05), capillary refilling time (P<0.01), neutrophil numbers (P<0.01), serum albumin (P<0.01), plasma total antioxidant status (P<0.01), and a lower rate of mitogen induced proliferation of lymphocytes (P<0.05), in comparison with CG. On day 7 only an increase in total serum protein (P<0.05) and serum globulins (P<0.001) was seen in the EG. No difference in serum cortisol concentration was found. Long distance transportation induced an acute phase response impairing the cell-mediated immune response. Clinical examinations, including assessing CRT and body weight loss, and the monitoring of redox balance may be useful in evaluating the impact of extensive transport events on horses. A better understanding of the link between transportation stress, the immune system and the acute phase response is likely to inform strategies for enhancing the welfare of transported horses.