Towards personalized medicine for the treatment of equine asthma.

Although horses with asthma share similar clinical signs, the heterogeneity of the disease in terms of severity, triggering factors, inflammatory profile, and pathological features has hindered our ability to define biologically distinct subgroups. The recognition of phenotypes and endotypes could enable the development of precision medicine, including personalized, targeted therapy, to benefit affected horses. While in its infancy in horses, this review outlines the phenotypes of equine asthma and discusses how knowledge gained from targeted therapy in human medicine can be applied to evaluate the potential opportunities for personalized medicine in equine asthma and to suggest avenues for research to advance this emerging field.

Current Understanding of Equine Gut Dysbiosis and Microbiota Manipulation Techniques: Comparison with Current Knowledge in Other Species.

Understanding the importance of intestinal microbiota in horses and the factors influencing its composition have been the focus of many studies over the past few years. Factors such as age, diet, antibiotic administration, and geographic location can affect the gut microbiota. The intra- and inter-individual variability of fecal microbiota in horses complicates its interpretation and has hindered the establishment of a clear definition for dysbiosis. Although a definitive causal relationship between gut dysbiosis in horses and diseases has not been clearly identified, recent research suggests that dysbiosis may play a role in the pathogenesis of various conditions, such as colitis and asthma. Prebiotics, probiotics, and fecal microbiota transplantation to modulate the horse's gastrointestinal tract may eventually be considered a valuable tool for preventing or treating diseases, such as antibiotic-induced colitis. This article aims to summarize the current knowledge on the importance of intestinal microbiota in horses and factors influencing its composition, and also to review the published literature on methods for detecting dysbiosis while discussing the efficacy of gut microbiota manipulation in horses.

The Microbiota and Equine Asthma: An Integrative View of the Gut-Lung Axis.

Both microbe-microbe and host-microbe interactions can have effects beyond the local environment and influence immunological responses in remote organs such as the lungs. The crosstalk between the gut and the lungs, which is supported by complex connections and intricate pathways, is defined as the gut-lung axis. This review aimed to report on the potential role of the gut-lung gut-lung axis in the development and persistence of equine asthma. We summarized significant determinants in the development of asthma in horses and humans. The article discusses the gut-lung axis and proposes an integrative view of the relationship between gut microbiota and asthma. It also explores therapies for modulating the gut microbiota in horses with asthma. Improving our understanding of the horse gut-lung axis could lead to the development of techniques such as fecal microbiota transplants, probiotics, or prebiotics to manipulate the gut microbiota specifically for improving the management of asthma in horses.

Effects of soaked hay on lung function and airway inflammation in horses with severe asthma.

BACKGROUND: Reducing inhaled dust particles improves lung function in horses with severe asthma. Soaked hay is commonly used by owners, but its efficacy in improving lung function and inflammation has not been documented. OBJECTIVES: To measure the effects of soaked hay and alfalfa pellets in horses with severe asthma. ANIMALS: Ten adult horses with severe asthma from a research colony. METHODS: Prospective controlled trial. Horses in clinical exacerbation were housed indoors and allocated to be fed either soaked hay (n = 5) or alfalfa pellets (n = 5) for 6 weeks. Soaked hay was immersed for 45 minutes and dried out hay was discarded between meals. Pulmonary function and clinical scores were measured before and after 2, 4, and 6 weeks. Tracheal mucus scores and bronchoalveolar lavages were performed before and after 6 weeks. Lung function was analyzed with a linear mixed model using log-transformed data. RESULTS: Lung resistance decreased from (median (range)) 2.47 (1.54-3.95) to 1.59 (0.52-2.10) cmH2 O/L/s in the pellets group and from 1.89 (1.2-3.54) to 0.61 (0.42-2.08) cmH2 O/L/s in the soaked hay group over the 6-week period for an average difference of 1.06 cmH2 O/L/s for pellets (95% confidence interval [95% CI]: 0.09-2.04, P = .03, not significant after correction) and 1.31 cmH2 O/L/s for soaked hay (95% CI: -0.23 to 2.85, P < .001, significant). CONCLUSION AND CLINICAL IMPORTANCE: Soaked hay can control airway obstruction in horses with severe asthma. The strict protocol for soaking and discarding dried-out hay in this study could however be considered too great of an inconvenience by owners.

Severe asthma in horses is associated with increased airway innervation.

BACKGROUND: Altered innervation structure and function contribute to airway hyperresponsiveness in human asthma, yet the role of innervation in airflow limitation in asthma in horses remains unknown. HYPOTHESIS: To characterize peribronchial innervation in horses with asthma. We hypothesized that airway innervation increases in horses with asthma compared with controls. ANIMALS: Formalin-fixed lung samples from 8 horses with severe asthma and 8 healthy horses from the Equine Respiratory Tissue Biobank. Ante-mortem lung function was recorded. METHODS: Blinded case-control study. Immunohistochemistry was performed using rabbit anti-s100 antibody as a neuronal marker for myelinating and non-myelinating Schwann cells. The number and cumulative area of nerves in the peribronchial region and associated with airway smooth muscle were recorded using histomorphometry and corrected for airway size. RESULTS: Both the number (median [IQR]: 1.87 × 10-5 nerves/µm2 [1.28 × 10-5 ]) and the cumulative nerve area (CNA; 1.03 × 10-3 CNA/µm2 [1.57 × 10-3 ]) were higher in the peribronchial region of horses with asthma compared with controls (5.17 × 10-6 nerves/µm2 [3.76 × 10-6 ], 4.14 × 10-4 CNA/µm2 [2.54 × 10-4 ], Mann-Whitney, P = .01). The number of nerves within or lining airway smooth muscle was significantly higher in horses with asthma (4.47 × 10-6 nerves/µm2 [5.75 × 10-6 ]) compared with controls (2.26 × 10-6 nerves/µm2 [1.16 × 10-6 ], Mann-Whitney, P = .03). CONCLUSIONS AND CLINICAL IMPORTANCE: Asthma in horses is associated with greater airway innervation, possibly contributing to airway smooth muscle remodeling and exacerbating severity of the disease.

Effects of concentrated fecal microbiota transplant on the equine fecal microbiota after antibiotic-induced dysbiosis.

Bacterial imbalances are observed in intestinal diseases and fecal microbiota transplantation (FMT) has been used to restore the intestinal microbiota of horses. However, there is evidence that the current methods proposed for FMT in horses have limited efficacy. The objective of this study was to concentrate the bacteria present in the donor stool by centrifugation, and to test the effect in horses with antibiotic-induced dysbiosis. One healthy 11-year-old horse was selected as a fecal donor and 9 horses were given trimethoprim sulfadiazine (TMS) for 5 days to induce dysbiosis. Horses received either a concentrated FMT (cFMT, n = 3), fresh unconcentrated FMT (fFMT, n = 3), or 10% glycerol solution (vehicle, VEH, n = 3) by nasogastric tube for 3 days. Fecal samples were collected on Days 0, 4, 9, 11, and 21 for microbiota analysis (Illumina sequencing). The TMS significantly changed the bacterial composition of horses' feces (D0 versus D4). The composition of the cFMT and fFMT recipient horses was significantly different after transplantation compared to after antibiotic-induced dysbiosis (D4 versus D11), whereas the microbiota of the vehicle recipients was not, indicating that both protocols induced transient changes. However, preparation of FMT solutions markedly changed the original composition present in the donor's feces, with significant enrichment of Escherichia genus in the cFMT. Individual susceptibility to restoration of the microbiota was observed in horses, similar to what is known for other species. Our results suggest that concentrating bacteria should not be recommended in preparation of FMT solutions and that further research is required to improve current methods recommended to perform FMT in horses.

Des déséquilibres bactériens sont observés dans les maladies intestinales et la transplantation de microbiote fécal (FMT) a été utilisée pour la restaurer le microbiote intestinal des chevaux. Cependant, que les méthodes actuelles proposées pour FMT chez les chevaux ont une efficacité limitée. L'objectif de cette étude était de concentrer les bactéries présentes dans les selles du donneur par centrifugation, et de tester leur effet chez des chevaux atteints de dysbiose induite par les antibiotiques. Un cheval sain de 11 ans a été sélectionné comme donneur fécal et 9 chevaux ont reçu du triméthoprime sulfadiazine (TMS) pendant cinq jours pour induire une dysbiose. Les chevaux ont reçu soit une FMT concentrée (cFMT, n = 3), une FMT fraîche non concentrée (fFMT, n = 3) ou une solution de glycérol à 10 % (véhicule, VEH, n = 3) par sonde naso-gastrique pendant 3 jours. Des échantillons fécaux ont été prélevés aux jours 0, 4, 9, 11 et 21 pour analyse du microbiote (séquençage Illumina). Le TMS a significativement modifié la composition bactérienne des matières fécales des chevaux (D0 versus D4). La composition des chevaux receveurs cFMT et fFMT était significativement différente après la transplantation par rapport à la dysbiose induite par les antibiotiques (D4 versus D11), alors que le microbiote des receveurs de véhicules ne l'était pas, indiquant que les deux protocoles induisaient des changements transitoires. Cependant, la préparation des solutions FMT a considérablement modifié la composition originale présente dans les matières fécales du donneur, avec un enrichissement significatif du genre Escherichia dans le cFMT. Une susceptibilité individuelle à la restauration du microbiote a été observée chez les chevaux, à l'instar de ce qui est connu chez d'autres espèces. Nos résultats suggèrent que la concentration des bactéries ne devrait pas être recommandée dans la préparation des solutions FMT et que des recherches supplémentaires sont nécessaires pour améliorer les méthodes actuelles recommandées pour effectuer la FMT chez les chevaux.(Traduit par les auteurs).

Acute diarrhea in horses: A multicenter Canadian retrospective study (2015 to 2019).

This study describes the clinicopathological findings, diagnostic approach, treatment, and factors associated with non-survival of diarrheic horses admitted to 4 Canadian university teaching hospitals between 2015 and 2019. A total of 300 horses, ≥1-year-old, with acute diarrhea were included and represented 1.6% (300/18 481; range: 0.7 to 3%) of admissions during that period, 70% of the horses survived to discharge. Testing for enteropathogens was limited to a single fecal culture for Salmonella spp. in most cases. An enteropathogen was identified in 14% (42/300) of the horses, but in the hospital with higher testing rates enteropathogens were detected in 29% (16/55) of cases. Neorickettsia risticii was the pathogen most frequently detected (31%, 32/102). Antimicrobial drugs and plasma were administered to 57 and 8% of the cases, respectively. Laminitis occurred in 24/298 (8%) of the horses. A multivariable regression model identified an association between non-survival of diarrheic horses and colic signs, increased heart rate, packed cell volume, creatinine concentration, and decreased total protein concentration. A standardized approach for pathogen detection in diarrheic horses is not consistent among Canadian veterinary teaching hospitals, and testing for known pathogens is limited. Signs of colic, severe dehydration, endotoxemia, and hypoproteinemia are associated with non-survival of diarrheic horses.

Diarrhée aiguë chez le cheval : une étude rétrospective canadienne multicentrique (2015 à 2019). Cette étude décrit les résultats clinicopathologiques, l'approche diagnostique, le traitement et les facteurs associés à la non-survie de chevaux diarrhéiques admis dans quatre hôpitaux universitaires canadiens entre 2015 et 2019. Un total de 300 chevaux, ≥1 an, atteints de diarrhée aiguë ont été inclus et représentaient 1,6 % (300/18 481; intervalle : 0,7 à 3 %) des admissions au cours de cette période. Soixante-dix pourcents des chevaux ont survécu jusqu'à leur congé. La recherche d'agents entéropathogènes était limitée à une seule culture fécale pour Salmonella spp. dans la plupart des cas. Un agent entéropathogène a été identifié chez 14 % (42/300) des chevaux, mais à l'hôpital avec des taux de dépistage plus élevés, des agents entéropathogènes ont été détectés dans 29 % (16/55) des cas. Neorickettsia risticii était l'agent pathogène le plus fréquemment détecté (31 %, 32/102). Des médicaments antimicrobiens et du plasma ont été administrés respectivement à 57 et 8 % des cas. Une fourbure est survenue chez 24/298 (8 %) des chevaux. Un modèle de régression multivarié a identifié une association entre la non-survie des chevaux diarrhéiques et les signes de coliques, l'augmentation de la fréquence cardiaque, l'hématocrite, la concentration de créatinine et la diminution de la concentration totale de protéines. Une approche normalisée pour la détection des agents pathogènes chez les chevaux diarrhéiques n'est pas uniforme dans les hôpitaux d'enseignement vétérinaires canadiens, et les tests pour les agents pathogènes connus sont limités. Des signes de coliques, de déshydratation sévère, d'endotoxémie et d'hypoprotéinémie sont associés à la non-survie des chevaux diarrhéiques.(Traduit par Dr Serge Messier).

Real-Time PCR Differential Detection of Neorickettsia findlayensis and N. risticii in Cases of Potomac Horse Fever.

Potomac horse fever (PHF) is an acute and potentially fatal enterotyphlocolitis of horses with clinical signs that include anorexia, fever, diarrhea, and laminitis. Its incidence is increasing despite a commercially available vaccine. PHF is caused by Neorickettsia risticii, and the recently rediscovered and classified N. findlayensis. PHF diagnosis is currently accomplished using serology or nested PCR. However, both methods cannot distinguish the two Neorickettsia species that cause PHF. Further, the current N. risticii real-time PCR test fails to detect N. findlayensis. Thus, in this study, two Neorickettsia species-specific real-time PCR assays based on Neorickettsia ssa2 and a Neorickettsia genus-specific real-time PCR assay based on Neorickettsia 16S rRNA gene were developed. The ssa2 real-time PCR tests differentiated N. findlayensis from N. risticii in the field samples for which infection with either species had been verified using multiple other molecular tests and culture isolation, and the 16S rRNA gene real-time PCR detected both Neorickettsia species in the samples. These tests were applied to new field culture isolates from three Canadian provinces (Alberta, Quebec, Ontario) and Ohio as well as archival DNA samples from suspected PHF cases to estimate the prevalence of N. findlayensis in different geographic regions. The results suggest that N. findlayensis frequently causes PHF in horses in Alberta and Quebec. The development of these tests will allow rapid, sensitive, and specific diagnosis of horses presenting with clinical signs of PHF. These tests will also enable rapid and targeted treatment and help develop broad-spectrum vaccines for PHF.

Characterization of fungal exposure and dectin-1 expression in healthy horses and horses with severe asthma.

OBJECTIVE: To quantify dectin-1 expression in bronchoalveolar lavage fluid (BALF), create polyclonal antibodies against equine dectin-1 and localize it in tissues, and quantify fungal exposure in pastured and stabled asthmatic and nonasthmatic horses. SAMPLES: BALF samples from 6 controls and 6 horses with severe asthma. Stored lung and nasal wash samples. PROCEDURES: Dectin-1 expression was quantified by quantitative PCR (qPCR). Purified peptide from equine dectin-1 was used to generate polyclonal antibodies and was confirmed with immunological testing. Fungal exposure was quantified in BALF samples by counting fungal-like intracellular particles in phagocytic cells, by qPCR quantification of the "universal" 18S rRNA fungal gene, and by quantifying 36 specific fungi in equine and dust samples using qPCR assays. RESULTS: Equine dectin-1 was localized in tissues and cells, and functional isoforms were upregulated significantly in BALF after stabling. Pastured horses from both groups had low levels of fungi in BALF, and there was a significant increase in some specific fungi, most notably for Eurotium amstelodami, Wallemia sebi, and Aspergillus niger after stabling. However, stabled asthmatic horses had fewer phagocytized particles, less 18S rRNA signal, and fewer specific fungi compared to nonasthmatic horses. CLINICAL RELEVANCE: Stabling increases exposure to fungi, but asthmatic horses had fewer fungi reaching their lower airways, presumably resulting from congestion and narrowing of the airways. Exposure to fungi could contribute to airway inflammation by increasing dectin-1 functional isoforms, and exposure to indoor molds should be avoided.

Bilateral thoracic radiographs increase lesion detection in horses with pneumonia or pulmonary neoplasia but do not bring any additional benefit for inflammatory or diffuse pulmonary disease.

Published studies describing the effects of bilateral radiographic projections on the detection of equine pulmonary lesions are currently lacking. The objectives of this retrospective, single center, observational study were to compare unilateral and bilateral thoracic radiographic projections for the detection of pulmonary lesions in a group of horses. Based on their clinical diagnosis, 167 adults and foals with bilateral thoracic radiographs were classified as having pneumonia (n = 88), inflammatory or diffuse pulmonary disease (n = 72), and pulmonary masses (n = 7). After an initial interrater repeatability test, right-to-left and left-to-right projections were anonymized and independently interpreted by a radiologist blinded to the clinical diagnosis. Scores were attributed for each pattern/lesion (alveolar, interstitial, bronchial, nodules/masses, cavitary lesions) and each quadrant. Agreement between scores from each projection was evaluated with Bland-Altman plots. Lesions identified on one side but not on the contralateral projection were considered discordant. There was no preferential lateralization of pulmonary lesions. The prevalence of discordance was 14.4%, 9.0%, and 4.2% for alveolar pattern, nodules/masses, and cavitary lesions, respectively. Up to nine horses (10.2%) with pneumonia could have been misdiagnosed. A pulmonary mass would have been missed in one case. For inflammatory or diffuse disease, discordance was slight, and the addition of contralateral projections had no impact on radiographic interpretation. In conclusion, in horses with pneumonia or neoplasia, bilateral projections, or adding at least one contralateral caudoventral view, increased the probability of identifying pulmonary lesions. In horses with inflammatory or diffuse disease, bilateral thoracic radiography provided no additional benefit.

Assessment of thrombin generation in horses using a calibrated automated thrombogram.

BACKGROUND: The amount of thrombin generated reflects the endogenous thrombin potential (ETP), which depends on the balance of pro- and anticoagulant factors. The calibrated automated thrombogram (CAT) allows for the direct measurement of thrombin generation during the clotting process. OBJECTIVES: (1) To describe the results of the CAT assay in horses, (2) to establish intra-assay and intra- and interindividual variation of thrombin generation in healthy horses, and (3) to compare in vitro low-molecular-weight heparin (LMWH) sensitivity between healthy and sick horses. The hypothesis for the last objective is that inhibition of thrombin generation in sick horses requires higher heparin concentrations. METHODS: The plasma of 10 healthy mixed breed horses was used for the determination of normal thrombin generation parameters (lag time, time to peak, peak thrombin concentration, and ETP). Five of the healthy horses were compared with five horses with systemic inflammatory response syndrome (SIRS). In vitro heparin sensitivity was determined using LMWH. RESULTS: The intra-assay variation was small (<5%) for all parameters. Relatively large intra- and interindividual variation were observed in healthy horses. Four of the five sick horses with SIRS had a thrombogram compatible with a hypercoagulable state. The in vitro heparin sensitivity test suggested decreased sensitivity to LMWH in hypercoagulable states. CONCLUSIONS: The CAT assay could detect coagulopathy in horses. In vivo experiments are needed to confirm that it can be used to monitor responses to LMWH therapy.

Species-Level Gut Microbiota Analysis after Antibiotic-Induced Dysbiosis in Horses.

All current studies have used Illumina short-read sequencing to characterize the equine intestinal microbiota. Long-read sequencing can classify bacteria at the species level. The objectives of this study were to characterize the gut microbiota of horses at the species level before and after trimethoprim sulfadiazine (TMS) administration and to compare results with Illumina sequencing. Nine horses received TMS (30 mg/kg) orally for 5 days twice a day to induce dysbiosis. Illumina sequencing of the V4 region or full-length PacBio sequencing of the 16S rRNA gene was performed in fecal samples collected before and after antibiotic administration. The relative abundance and alpha diversity were compared between the two technologies. PacBio failed to classify the equine intestinal microbiota at the species level but confirmed Bacteroidetes as the most abundant bacteria in the feces of the studied horses, followed by Firmicutes and Fibrobacteres. An unknown species of the Bacteroidales order was highly abundant (13%) and deserves further investigation. In conclusion, PacBio was not suitable to classify the equine microbiota species but detected greater richness and less unclassified bacteria. Further efforts in improving current databanks to be used in equine studies are necessary.

Nicoletella semolina in the airways of healthy horses and horses with severe asthma.

BACKGROUND: Nicoletella semolina was identified in the airways of horses and its low prevalence could be because of its difficult differentiation from other Pasteurellaceae. OBJECTIVES: To develop a molecular method for the identification of N. semolina and to evaluate its prevalence in the mouth and the airways of healthy and severe asthmatic horses. ANIMALS: Six healthy and 6 severely asthmatic horses in phase I, 10 severely asthmatic horses in phase II, and 10 healthy horses in phase III. METHODS: Cohort (phases I and II) and cross-sectional (phase III) studies. Quantitative polymerase chain reaction primers targeting the sodA gene were optimized. N. semolina was quantified in oral and nasal washes and in bronchoalveolar lavage fluid (BALF; phase I, sampled twice), in nasal washes and BALF (phase II, sampled twice), and in nasal washes (phase III). RESULTS: N. semolina was found in the nose of 5, 10, and 9 horses in phases I, II, and III, respectively (first sampling for phases I and II). Six BALF from 5 different horses were positive for N. semolina in phase II. In phase I, there was no significant difference in the nasal loads of healthy horses (median (range): 2.04 × 104 copies/mL (0-2.44 × 105 )) and asthmatic horses in exacerbation (3.75 × 102 (0-4.84 × 106 ); Wilcoxon's rank sum test, P = .57). CONCLUSIONS AND CLINICAL IMPORTANCE: N. semolina is commonly found in the airways of horses. The potential pathogenicity of N. semolina remains to be elucidated, but the molecular technique we developed will facilitate future studies.

Severe asthma is associated with a remodeling of the pulmonary arteries in horses.

Pulmonary hypertension and cor pulmonale are complications of severe equine asthma, as a consequence of pulmonary hypoxic vasoconstriction. However, as pulmonary hypertension is only partially reversible by oxygen administration, other etiological factors are likely involved. In human chronic obstructive pulmonary disease, pulmonary artery remodeling contributes to the development of pulmonary hypertension. In rodent models, pulmonary vascular remodeling is present as a consequence of allergic airway inflammation. The present study investigated the presence of remodeling of the pulmonary arteries in severe equine asthma, its distribution throughout the lungs, and its reversibility following long-term antigen avoidance strategies and inhaled corticosteroid administration. Using histomorphometry, the total wall area of pulmonary arteries from different regions of the lungs of asthmatic horses and controls was measured. The smooth muscle mass of pulmonary arteries was also estimated on lung sections stained for α-smooth muscle actin. Reversibility of vascular changes in asthmatic horses was assessed after 1 year of antigen avoidance alone or treatment with inhaled fluticasone. Pulmonary arteries showed increased wall area in apical and caudodorsal lung regions of asthmatic horses in both exacerbation and remission. The pulmonary arteries smooth muscle mass was similarly increased. Both treatments reversed the increase in wall area. However, a trend for normalization of the vascular smooth muscle mass was observed only after treatment with antigen avoidance, but not with fluticasone. In conclusion, severe equine asthma is associated with remodeling of the pulmonary arteries consisting in an increased smooth muscle mass. The resulting narrowing of the artery lumen could enhance hypoxic vasoconstriction, contributing to pulmonary hypertension. In our study population, the antigen avoidance strategy appeared more promising than inhaled corticosteroids in controlling vascular remodeling. However, further studies are needed to support the reversibility of vascular smooth muscle mass remodeling after asthma treatment.

Fecal microbiota in horses with asthma.

BACKGROUND: Gastrointestinal microbiota can be influenced by several factors, including diet and systemic inflammation, and in turn could act as a modulator of the allergic response. Fecal microbiota of horses with asthma has not been described. HYPOTHESIS/OBJECTIVES: Analyze the bacterial fecal microbiota of horses with and without asthma under different environment and diet conditions, during both remission and exacerbation. METHODS: Prospective observational study. Feces from 6 asthmatic and 6 healthy horses were collected under 3 different conditions: on pasture, housed indoors receiving good quality hay ("good hay"), and housed indoors receiving poor quality hay ("dusty hay"). Sequencing was performed using an Illumina MiSeq platform and data were processed using the software mothur v.1.41.3 and LEfSe. RESULTS: In horses with asthma, low-abundance bacteria were more affected by changes in environment and diet (ie, when horses were experiencing an exacerbation), as shown by changes in membership and results from the LEfSe analysis. There was a significant increase in the relative abundance of Fibrobacter in healthy horses eating hay, a change that was not observed in horses with asthma. CONCLUSIONS AND CLINICAL IMPORTANCE: The intestinal microbiota of horses with asthma does not adapt in the same way to changes in diet and environment compared to the microbiota of healthy horses. Mechanisms explaining how airway obstruction and inflammation could influence the intestinal microbiota and how in turn this microbiota could modulate systemic inflammation in asthmatic horses deserves further investigation.

Acute, subacute and chronic sequelae of horses accidentally exposed to monensin-contaminated feed.

BACKGROUND: Monensin is highly toxic to horses and inadvertent ingestion can result in cardiac injury and death. OBJECTIVES: To describe sequelae of monensin ingestion and to determine clinical predictors of outcome. STUDY DESIGN: Observational clinical study. METHODS: Physical examination, electrocardiogram and echocardiography were performed on 76 horses accidentally exposed to monensin-contaminated feed. Four horses were examined within 14 days of exposure (acute period), 29 horses were examined between 15 and 45 days post-exposure (subacute period) and 70 horses were examined 4-10 months after exposure (chronic period). Follow-up information was obtained for 56 horses by telephone interviews approximately 16 months after exposure. RESULTS: Cardiac abnormalities were detected in 4/4, 19/29 and 31/70 horses during the acute, subacute and chronic periods, respectively. Sixteen months post-exposure, 34 of the 64 horses (53%) for which the outcome was known had returned to their previous use, 13 (20%) were reported to be exercise intolerant, three (5%) were retired and 14 (22%) were dead (two deaths, 12 euthanasia). Thinning of the myocardium observed at any point in time was associated with a negative outcome. Heterogeneity of the myocardium observed in the acute/subacute period was associated with a negative outcome while subjective contractile intraventricular dyssynchrony, cardiac chamber dilation, decreased fractional shortening and multiple premature ventricular complexes observed in the chronic period were associated with a negative outcome. Some horses with significant changes associated with a negative outcome in the chronic phase still returned to their previous work. MAIN LIMITATIONS: No control group and only 27 horses were examined more than once. CONCLUSIONS: Clinical outcome of horses exposed to sublethal doses of monensin is highly variable. The presence of heterogeneity and thinning of the myocardium shortly after intoxication were associated with a negative outcome.

Tracheal microbial populations in horses with moderate asthma.

BACKGROUND: There are limited data on potential dysbiosis of the airway microbiota in horses with asthma. HYPOTHESIS/OBJECTIVES: We hypothesized that the respiratory microbiota of horses with moderate asthma is altered. Our objectives were (a) to quantify tracheal bacterial populations using culture and qPCR, (2) to compare aerobic culture and qPCR, and (c) to correlate bacterial populations with bronchoalveolar lavage fluid (BALF) cytology. ANIMALS: Eighteen horses with moderate asthma from a hospital population and 10 controls. METHODS: Prospective case-control study. Aerobic culture was performed on tracheal aspirates, and streptococci, Pasteurella multocida, Chlamydophila spp., Mycoplasma spp., as well as 16S (bacterial) and 18S (fungal) rRNA subunits were quantified by qPCR. RESULTS: Potential pathogens such as Streptococcus spp., Actinobacillus spp., and Pasteurellaceae were isolated from 8, 5, and 6 horses with asthma and 3, 0, and 2 controls, respectively. There was a positive correlation between Streptococcus spp. DNA and 16S rRNA gene (r ≥ 0.7, P ≤ 0.02 in both groups), but the overall bacterial load (16S) was lower in asthma (1.5 ± 1.3 versus 2.5 ± 0.8 × 104 copy/µL, P < 0.05). There was no association between microbial populations and clinical signs, tracheal mucus or BALF inflammation. CONCLUSIONS AND CLINICAL IMPORTANCE: This study does not support that bacterial overgrowth is a common feature of chronic moderate asthma in horses. Lower bacterial load could suggest dysbiosis of the lower airways, either as a consequence of chronic inflammation or previous treatments, or as a perpetuating factor of inflammation.

Cryptococcus gattii pneumonia in an adult horse which had travelled in an endemic area.

A gelding from eastern Canada was presented for cough and exercise intolerance 14 months after it had travelled on Vancouver Island. Cryptococcus gattii pneumonia was diagnosed based on cytology, antigen titers, and polymerase chain reaction (PCR). The horse was treated with fluconazole for 10 months. Delayed C. gattii infection can occur after travel in an endemic area.

Pneumonie à Cryptococcus gattii chez un cheval adulte ayant voyagé dans une région endémique. Un cheval hongre de l'est canadien fut présenté pour de la toux et de l'intolérance à l'exercice 14 mois après avoir voyagé sur l'Île de Vancouver. Une pneumonie à Cryptococcus gattii fut diagnostiquée sur la base de la cytologie, des titres antigéniques, et de la réaction d'amplification en chaîne par la polymérase (PCR). Le cheval fut traité avec du fluconazole pendant 10 mois. Une infection à retardement par C. gattii peut survenir à la suite d'un voyage dans une région endémique.(Traduit par Dr Serge Messier).

Investigation of blood biomarkers for the diagnosis of mild to moderate asthma in horses.

BACKGROUND: Asthma in horses is associated with nonspecific respiratory clinical signs and may be manifested only as exercise intolerance. Its diagnosis relies on bronchoalveolar lavage fluid (BALF) cytology in the presence of compatible clinical signs. The identification of blood biomarkers for this condition would facilitate diagnosis in the field, because there are regional areas where BAL is not routinely performed in clinical practice. OBJECTIVE: Identification of blood biomarkers for the diagnosis of asthma in horses. ANIMALS: Fourteen horses with asthma with increased neutrophil numbers in BALF (neutrophilic asthma), 9 healthy control horses, and 10 horses with other pathologic conditions (pathologic controls). METHODS: Physical examination, clinical score, hematology, and BALF cytology (in a subset of horses) were performed. Serum concentrations of surfactant protein D (SP-D), haptoglobin, and secretoglobin (SCGB) were measured using commercial ELISA assays. RESULTS: Serum concentration of SP-D > 43 ng/mL, serum concentration of haptoglobin >5730 ng/mL, and serum concentration of SCGB <19 ng/mL allowed differentiation of horses with neutrophilic asthma from horses of the control groups (healthy and pathologic) with sensitivity of 55, 95, and 75%, and specificity of 67, 28, and 60%, respectively. Specificity of 100% and sensitivity of 45% were obtained with the combination of SP-D, haptoglobin, and SCGB at the serum concentrations indicated above. Specificity of 95% and sensitivity of 45% were obtained with the combination of SP-D and SCGB serum concentrations. CONCLUSIONS AND CLINICAL IMPORTANCE: Haptoglobin, SCGB, and SP-D may be diagnostic aids in horses with clinical signs of lower airway disease and neutrophilic pulmonary inflammation.

Modified stall-side crossmatch for transfusions in horses.

BACKGROUND: After-hours or out-of-clinic crossmatches are often limited by the lack of access to specialized material and technical expertise. HYPOTHESIS/OBJECTIVES: The goal was to adapt a stall-side crossmatch test for pretransfusion evaluation in horses. ANIMALS: Twelve healthy mares (plasma and blood donors, teaching mares). METHODS: In a prospective study, blood from 12 mares was used to compare the results of 132 crossmatches performed with a rapid gel assay to crossmatches performed with a microgel column assay, and with predicted compatibilities based on blood types and detection of antibodies at a reference laboratory (microplate assay). The rapid gel assay protocol for dogs was adapted to decrease the formation of rouleaux that initially precluded equine erythrocytes migration through the gel. RESULTS: There was a good agreement between the rapid gel assay and the microgel assay as well as with the predicted compatibilities (κ > .6 for both). Agreement was higher between the microgel assay and the predicted compatibilities (κ = .8). The rapid gel assay failed to detect 6 predicted Aa incompatibilities (agglutinins-related), 3 of which were also not detected with the microgel assay. CONCLUSIONS AND CLINICAL IMPORTANCE: Based on these results, the modified rapid gel assay could be useful in settings when access to the microgel assay is not available. Discrepancies between both gel techniques and predicted compatibilities were most often low-grade agglutination, which warrants further investigation to assess their clinical importance.