Evaluation of a prototype dynamic laryngoplasty system in vitro with an equine vacuum airflow system.

OBJECTIVE: To evaluate a prototype dynamic laryngoplasty system (DLPS) in a static airflow model. STUDY DESIGN: Experimental. SAMPLE POPULATION: Ten equine larynges. METHODS: The right arytenoid was fixed in abduction in all specimens. A left-sided laryngoplasty was performed with No. 2 Fiberwire and a FASTakII anchor. Each larynx was tested in a static airflow model. The system was adjusted to a flow rate of 55 L/s and prelaryngeal pressure of 12 mm Hg prior to testing in maximal arytenoid abduction. In phase 1, the left suture was loosened, shortened, and tested in 3-mm steps from 0 to 30 mm. In phase 2, the suture was tied with the DLPS in position at a target left-to-right quotient angle (LRQ) of 0.5. The DLPS was activated to target psi of 0, 25, and 50 for testing. Translaryngeal impedance (TLI), LRQ, cross-sectional areas (CSA), and resultant change in LRQ and CSA between, before, and during airflow testing were calculated. RESULTS: In phase 1, TLI was reduced by suture shortening up to 6 mm (P = .001) but not by additional shortening (P > .05). In phase 2, activation of the DLPS reduced the TLI from 0 psi (0.43 ± 0.08 mm Hg/L/s) to 25 psi (0.16 ± 0.04 mm Hg/L/s, P < .001), but no further reduction was detected at maximal psi (P = .10). CONCLUSION: Activation of the DLPS effectively reduced TLI. CLINICAL SIGNIFICANCE: These results justify further investigation of the DLPS to assess its clinical applicability.

In vitro evaluation of the effect of a prototype dynamic laryngoplasty system on arytenoid abduction.

OBJECTIVE: To determine the effect of a prototype dynamic laryngoplasty system (DLPS) on arytenoid abduction. STUDY DESIGN: In vitro experimental. STUDY POPULATION: Ten equine larynges. METHODS: Dissected larynges were mounted, and the right arytenoid was maximally abducted for testing. A left-sided laryngoplasty (LP) was performed by using a strand of No. 2 FiberWire and a FASTakII anchor. Phase 1 involved tightening the suture, without the DLPS device in place, in 1-mm increments and acquiring a digital image of the rima glottidis at each increment. Phase 2 involved tying the suture with the DLPS in place at a left to right quotient (LRQ) of 0.7. Digital images were subsequently taken at 3 stages of DLPS activation (0, 25, and 50 or maximal psi) and analysed to calculate LRQ. RESULTS: All tests were completed for 9 larynges. In phase 1, a total shortening of 25.89 ± 1.27 mm was possible, which increased the LRQ from 0.59 ± 0.02 to 1.07 ± 0.12. In phase 2, activation of the DLPS increased the LRQ from 0.70 ± 0.05 to 0.97 ± 0.09. This change in LRQ equated to 18.7 mm of shortening on the basis of phase 1 results. The maximum psi of the DLPS achieved was 37.33 ± 5.96. CONCLUSION: The DLPS increased the degree of arytenoid abduction in vitro. This change in LRQ equated to 18.7 mm of shortening of the LP suture based on phase 1 results. CLINICAL IMPACT: These results support further evaluation of the DLPS to determine the effect of changes in DLPS on airway resistance.

In vitro comparison of 3 techniques of prosthesis attachment to the muscular process of the equine arytenoid cartilage.

OBJECTIVE: To determine the degree and ease of arytenoid abduction achieved with abaxial placement of a FASTak II suture anchor compared to 2 suture patterns with different directions of insertion-caudomedial to craniolateral and medial to lateral. STUDY DESIGN: Ex vivo experimental. STUDY POPULATION: Cadaveric larynges from 10 Standardbred racehorses. METHODS: Each larynx was sequentially instrumented with all 3 arytenoid suture attachment in random order: (1) abaxial placement of a FASTak II suture, (2) caudomedial to craniolateral suture, and (3) medial to lateral suture placement. Each construct was abducted at 5N increments from 0 to 25N and the left to right quotient angle ratio (LRQ) measured from digital pictures acquired at each sequential increment. RESULTS: Arytenoid abduction (higher LRQ) was greater with FASTak II construct than either of the suture patterns. The largest difference occurred at 5N. Approximately 50% less force was required to achieve an LRQ of 1.0 with the FASTak II anchor compared to the suture patterns. No difference was detected between the 2 suture constructs throughout the study. CONCLUSION: Use of the FASTak II suture anchor improved arytenoid abduction compared to 2 suture patterns and minimized the suture loads required to achieve maximum arytenoid abduction. CLINICAL RELEVANCE: Use of the FASTak II anchor may decrease the suture load required to achieve arytenoid abduction in clinical cases. This may reduce the load placed on the laryngoplasty, thereby, minimizing postoperative loss of abduction.

Standing left recurrent laryngeal neurectomy for prospective evaluation of laryngeal hemiplegia evaluated by a high-speed treadmill test.

OBJECTIVE: To describe left recurrent laryngeal neurectomy (LRLn) performed under standing sedation and evaluate the effect of LRLn on upper respiratory tract function using a high-speed treadmill test (HST). We hypothesized that (1) unilateral LRLn could be performed in standing horses, resulting in ipsilateral arytenoid cartilage collapse (ACC); and (2) HST after LRLn would be associated with alterations in upper respiratory function consistent with dynamic ACC. ANIMALS: 6 Thoroughbred horses. METHODS: The horses were trained and underwent a baseline HST up to 14 m/s at 5% incline until fatigue. Evaluation included; airflow, pharyngeal and tracheal pressures, and dynamic upper respiratory tract endoscopy. Trans-laryngeal impedance (TLI) and left-to-right quotient angle ratio (LRQ) were calculated after testing. The following day, standing LRLn was performed in the mid-cervical region. A HST was repeated within 4 days after surgery. RESULTS: Standing LRLn was performed without complication resulting in Havemayer grade 4 ACC at rest (complete paralysis) and Rakestraw grade C or D ACC (collapse up to or beyond rima glottis midline) during exercise. Increasing treadmill speed from 11 to 14 m/s increased TLI (P < .001) and reduced LRQ (P < .001). Neurectomy resulted in an increase in TLI (P = .021) and a reduction in LRQ (P < .001). CLINICAL RELEVANCE: Standing LRLn induces laryngeal hemiplegia that can be evaluated using a HST closely after neurectomy. Standing LRLn may be useful for future prospective evaluations of surgical interventions for laryngeal hemiplegia.

Heritability of sudden cardiac death in Thoroughbred racehorses.

BACKGROUND: Sudden cardiac death (SCD) is an important cause of exercise-associated fatalities in Thoroughbred racehorses. Equine deaths share similarities with fatalities in human athletes that result from inherited cardiac disease. Whilst genetic causes have been postulated in horses, these have not been confirmed and heritability of SCD has not previously been estimated in Thoroughbred racehorses. OBJECTIVES: To determine the heritability of SCD in a sample population of Thoroughbred racehorses. STUDY DESIGN: Retrospective case-control study. METHODS: Steward and post-mortem reports of Thoroughbred racehorses in Australia between 2007 and 2020 were reviewed to identify horses with SCD. Control horses were randomly selected from races in which SCD occurred or from races on the date of the case fatality. A five-generation integrated pedigree chart was collected for each horse. Estimates of heritability were obtained using an animal model in the ASReml-R program with variance components estimated assuming SCD was normally distributed, and on the logit transformed scale. Inbreeding coefficients were calculated and the risk of producing SCD-affected progeny was calculated for stallions that sired ≥5 individuals in the case-control population. RESULTS: Ninety-three horses with SCD and 465 control horses were identified. Heritability on the underlying scale was 0.15 ± 0.09 (logit animal) and 0.24 ± 0.12 (normal animal). Inbreeding coefficients were not significantly different between groups. Of the 16 first generation sires that appeared ≥5 times in the case-control data set, two sires more frequently produced affected progeny (OR 7.95-10.41). MAIN LIMITATIONS: Challenges in definitively confirming SCD may lead to misclassification of some cases. Some control horses may have not been exposed to environmental influences of SCD. Case numbers are low and the studied population may not represent the entire Thoroughbred genetic pool. CONCLUSION: The heritability of SCD in this population was relatively low. However, individual stallions appear more likely to produce affected progeny. Further studies are required to understand the genetic and environmental influences that contribute to disease expression.

Yearling laryngeal function in Thoroughbreds that underwent a laryngoplasty differs from controls.

BACKGROUND: Yearling laryngeal function (YLF) is frequently assessed at the time of sale and the outcomes of these assessments can have significant economic implications. The YLF of horses that subsequently underwent a prosthetic laryngoplasty (PL) is unknown. OBJECTIVES: We hypothesised horses with YLF ≥grade II.2 would be at increased risk of requiring PL, compared with YLF

Implementation of a prototype dynamic laryngoplasty system in standing sedated horses provided arytenoid abduction control at seven days postoperatively.

OBJECTIVE: Evaluate a prototype dynamic laryngoplasty system (DLPS) in horses; a feasibility study. ANIMALS: 7 healthy Standardbred adult horses. METHODS: This was an in vivo experimental study. Horses had a standing surgical procedure to induce complete laryngeal hemiplegia, which was subsequently treated using the dynamic laryngoplasty system (DLPS). Activation of the DLPS was achieved using an injection port exiting through the skin (n = 2) or a subcutaneous injection port (n = 5). For each horse, endoscopic examinations of the upper respiratory tract were performed preoperatively, intraoperatively, and 7 days postoperatively. Left-to-right quotient ratios calculated during inactivated and activated states were obtained from still images of the rima glottidis acquired during day 7. In 3 horses, the device was intentionally overinflated to evaluate for device failure, and postmortem examinations were performed on day 7. For the remaining 4 horses, upper respiratory tract endoscopy was repeated at 1 month postoperatively, with no subsequent postmortem exam. RESULTS: No perioperative complications occurred, and the DLPS was effectively delivered in all horses under standing sedation. The left-to-right quotient ratio at day 7 postoperatively could be altered from a resting position of 0.76 (± 0.06) to a maximum of 0.97 (± 0.06; P < .05). The degree of arytenoid abduction could not be significantly altered after 1 month of device implantation, suspected to be due to peri-implant fibrosis. No coughing nor tracheal contamination was observed at all time points or during inflation. CLINICAL RELEVANCE: The ability to alter the degree of abduction at 7 days postoperatively with the DLPS may be beneficial in selective cases.

Assessment of P Wave Indices in Healthy Standardbred Horses.

P wave indices are used as non-invasive electrocardiographic markers of atrial remodelling in humans. Few studies have investigated their use in animals. The aim of this study was to measure P wave duration and P wave dispersion (Pd) in healthy standardbred horses and investigate variables that might influence these measurements. A 12-lead electrocardiogram was recorded at rest and P wave indices were calculated in 53 horses. A general linear model was used to investigate the main effects: age, bodyweight, sex, resting heart rate, presence of a murmur, exercise status and the number of years raced. There were significant associations with exercise status for both the maximum P wave duration and Pd, with both values being increased in strenuously exercising versus non-active horses. Furthermore, a significant moderate positive correlation was identified between the duration of exercise (number of years raced) and both Pmax and Pd. No other significant associations were identified. These findings are similar to those reported in elite human athletes versus sedentary individuals. The increases in these P wave indices most likely occur due to prolongation and heterogeneity in atrial conduction time, which are associated with structural and electrical remodelling, and may explain the increased risk of atrial fibrillation in athletic horses.

Heart rate variability during high-speed treadmill exercise and recovery in Thoroughbred racehorses presented for poor performance.

BACKGROUND: Heart rate variability (HRV) analysis measures the inter-beat interval variation of successive cardiac cycles. Measurement of these indices has been used to assess cardiac autonomic modulation and for arrhythmia identification in exercising horses. OBJECTIVES: To report HRV indices during submaximal exercise, strenuous exercise and recovery, and explore relationships with clinical conditions (arrhythmias, lameness, equine gastric ulcer syndrome [EGUS], lower airway inflammation and upper respiratory tract obstructions [URTOs]) in Thoroughbred racehorses. STUDY DESIGN: Retrospective, observational cross-sectional study. METHODS: One hundred and eighty Thoroughbred horses underwent a treadmill exercise test with simultaneous electrocardiographic recording. Time-domain HRV indices (standard deviation of the R-R interval [SDRR]; root mean square of successive differences [RMSSD]) were derived for submaximal and strenuous exercise and recovery segments. Clinical conditions (arrhythmia [during each phase of exercise], lameness, EGUS, lower airway inflammation and URTO) were assigned to binary categories for statistical analysis. Relationships between selected HRV indices and the clinical conditions were explored using linear regression models. RESULTS: During submaximal exercise, lameness was associated with decreased logRMSSD (B = -0.19 95% confidence interval [CI] -0.31 to -0.06, p = 0.006) and arrhythmia was associated with increased logRMSSD (B = 0.31 95% CI 0.01-0.608, p = 0.04). During strenuous exercise, arrhythmia was associated with increased HRV indices (logSDRR B = 0.51 95% CI 0.40-0.62, p < 0.001; RMSSD B = 0.60 95% CI 0.49-0.72, p < 0.001). During recovery, arrhythmia was associated with increased HRV indices (logSDRR B = 0.51 95% CI 0.40-0.62, p < 0.001, logRMSSD B = 0.60 95% CI 0.49-0.72, p < 0.001). MAIN LIMITATIONS: The main limitations of this retrospective study were that not every horse had the full range of clinical testing, therefore some horses may have had undetected abnormalities. CONCLUSIONS: The presence of arrhythmia increased HRV in both phases of exercise and recovery. Lameness decreased HRV during submaximal exercise.

Identification and kinetics of microsomal and recombinant equine liver cytochrome P450 enzymes responsible for in vitro metabolism of omeprazole.

In humans, omeprazole is metabolised by cytochrome P450 (CYP450) CYP2C19 and CYP3A4 with differences in CYP2C19 genotypes leading to variable response to therapy. Despite a wide use of omeprazole in horses with evidence of variable therapeutic efficiency, information regarding enzymatic metabolism is not currently available. This study aims to describe the in vitro kinetics of omeprazole metabolism and determine which enzyme(s) are responsible for omeprazole metabolism in horses. Omeprazole (0-800 uM) was incubated with liver microsomes and a panel of equine recombinant CYP450s (eq-rCYP). Metabolite concentrations were quantified by LC-MS and the kinetics of metabolites' formation were calculated by non-linear regression analysis. The in vitro liver microsomes formed three metabolites (5-hydroxy-omeprazole, 5-O-desmethyl-omeprazole and omeprazole-sulfone). The 5-O-desmesthyl-omeprazole formation was best fitted to a two enzyme Michaelis-Menten (MM) model with the high affinity site Clint double that of the low affinity site. For 5-hydroxy-omeprazole the best fit was to a 1 enzyme MM model with a Clint higher than for 5-O-desmesthyl-omeprazole (0.12 vs 0.09 pmol/min/pmol P450). The formation of omeprazole-sulfone was negligible. Recombinant CYP3A89 and CYP3A97 produced substantial amounts of 5-hydroxy-omeprazole (1551.72 ng/mL and 1665.33 ng/mL, respectively), while 5-O-desmethyl-omeprazole and omeprazole-sulfone were formed to a much lesser extent by multiple eq-rCYP from the CYP2C and CYP3A family. In vitro metabolism of omeprazole in horses is different to that in humans, with major metabolites produced by the CYP3A family. The current study provides the basis for further investigations of CYP450 single nucleotide polymorphisms that could affect omeprazole metabolism and therapeutic efficacy.

Associations between postrace atrial fibrillation and measures of performance, racing history and airway disease in horses.

BACKGROUND: Atrial fibrillation (AF) is the most common performance limiting arrhythmia in racehorses. High dose exercise and airway disease promote AF in humans. Few studies have investigated epidemiological factors associated with AF in horses. OBJECTIVES: Explore variables relating to performance, exercise volume and postrace endoscopic findings in horses with AF. ANIMALS: A total of 164 horses with poor race performance and postrace AF were compared to 321 horses with poor performance without AF (PP) and 314 horses performing to expectation (TE). METHODS: Horse-level and race-level variables for horses racing in Australia and Hong Kong from 2009 to 2021 were compared using univariable and multivariable logistic regression. Postrace endoscopic exercise-induced pulmonary hemorrhage (EIPH) and tracheal mucus accumulation (TMA) grades for AF and PP horses were compared using chi-squared analysis. RESULTS: Variables that were significant in the multivariable model of AF compared to TE were distance (lengths) behind the winner, (odds ratio [OR]; 95% confidence interval [95% CI], 1.41 [1.32-1.51], P < .0001), cumulative prize money per start before the event (OR [95% CI] 1.02 [1.01-1.03], P = .01) and age (OR [95% CI] 0.72 [0.55-0.92], P = .01). More AF horses had EIPH grade ≥ 3 (23/109; 21.1%) than PP horses (7/213; 3.3%; OR [95%CI] 7.9 [3.3-20.2], P < .0001). CONCLUSIONS AND CLINICAL IMPORTANCE: Acute race performance was substantially impaired by AF but career earnings before the event were not inferior. Exercise volume did not promote AF. Higher grades of EIPH found in AF horses suggests a mechanistic relationship between these conditions.

Evaluation of recurrent laryngeal neuropathy in domestic and feral horse populations in Australia using histologic and immunohistochemical analysis: A pilot study.

BACKGROUND: Little is known about potential differences in the left recurrent laryngeal nerve (Lrln) and left cricoarytenoideus dorsalis (LCAD) muscle between domestic and feral horse populations. If a difference exists, feral horses may provide a useful control population for research related to recurrent laryngeal neuropathy (RLN) and increase our understanding of potential population pressures influencing the incidence RLN. OBJECTIVES: The objective of this study was to compare the Lrln and LCAD of domestic and feral horses using histological and immunohistochemical techniques (IHC). METHODS: Sixteen horses, domestic (n = 8) and feral (n = 8), without clinical or ancillary examinations that were processed at an abattoir had the Lrln and LCAD muscle harvested immediately following death. Carcass weights were recorded. Subjective and morphometric histologic assessment were performed on Lrln sections. The LCAD was assessed for myosin heavy chain (fibre type proportion, diameter and grouping using IHC. RESULTS: Fibre-type grouping consistent with RLN was seen in both groups. Regenerating fibre clusters were more common in domestic compared to feral horses (p = 0.04). No other histologic differences occurred between groups. Muscle fibre typing demonstrated a lower mean percentage of type IIX fibres in the feral group compared to the domestic group (p = 0.03). There was no difference in type I or IIA proportions or mean diameter of any fibre type between the groups. CONCLUSIONS: The domestic population showed evidence of nerve regeneration suggesting RLN in this group, yet this was not supported by the higher proportion of type IIX muscle fibres compared to the feral population. Further evaluation to clarify the significance and wider occurrence of the differences is indicated.

Contribution of the Oral and Gastrointestinal Microbiomes to Bloodstream Infections in Leukemia Patients.

Bloodstream infections (BSIs) pose a significant mortality risk for acute myeloid leukemia (AML) patients. It has been previously reported that intestinal domination (>30% relative abundance [RA] attributed to a single taxon) with the infecting taxa often precedes BSI in stem cell transplant patients. Using 16S rRNA amplicon sequencing, we analyzed oral and stool samples from 63 AML patients with BSIs to determine the correlation between the infectious agent and microbiome composition. Whole-genome sequencing and antimicrobial susceptibilities were performed on all BSI isolates. Species-level detection of the infectious agent and presence of antibiotic resistance determinants in the stool (blaCTX-M-15, blaCTX-M-14, cfrA, and vanA) were confirmed via digital droplet PCR (ddPCR). Individuals with Escherichia coli (stool P < 0.001), Pseudomonas aeruginosa (oral P = 0.004, stool P < 0.001), and viridans group streptococci (VGS) (oral P = 0.001) bacteremia had a significantly higher relative abundance of those respective genera than other BSI patients, which appeared to be site specific. Although 78% of patients showed presence of the infectious genera in the stool and/or saliva, only 7 exhibited microbiome domination. ddPCR confirmed species specificity of the 16S data and detected the antibiotic resistance determinants found in the BSI isolates within concurrent stools. Although gastrointestinal (GI) domination by an infecting organism was not present at the time of most BSIs in AML, the pathogens, along with AMR elements, were detectable in the majority of patients. Thus, rapid genetic assessment of oral and stool samples for the presence of potential pathogens and AMR determinants might inform personalized therapeutic approaches in immunocompromised patients with suspected infection. IMPORTANCE A major cause of mortality in hematologic malignancy patients is BSI. Previous studies have demonstrated that bacterial translocation from the GI microbiome is a major source of BSIs and is often preceded by increased levels of the infectious taxa in the GI (>30% abundance by 16S rRNA sequencing). In this study, we sought to better understand how domination and abundance levels of the oral and gut microbiome relate to bacteremia occurrence in acute myeloid leukemia patients. We conclude that analyses of both oral and stool samples can help identify BSI and antimicrobial resistance determinants, thus potentially improving the timing and tailoring of antibiotic treatment strategies for high-risk patients.

Horse Grimace Scale Does Not Detect Pain in Horses with Equine Gastric Ulcer Syndrome.

Equine gastric ulcer syndrome (EGUS) is a highly prevalent and presumptively painful condition, although the amount of pain horses might experience is currently unknown. The aims of this study were to determine if the Horse Grimace Scale (HGS) could identify pain behaviours in horses with and without EGUS and if severity would be positively associated with the HGS score. Horse grimace scale scores were assessed blindly using facial photographs by seven observers and involved evaluation of 6 facial action units as 0 (not present), 1 (moderately present) and 2 (obviously present). Lameness examination, serum amyloid A (SAA) measurement and gastroscopy evaluation were performed on all horses. Horses (n = 61) were divided into two and three groups based on the presence (yes, no) and severity (none, mild, moderate-severe) of EGUS, respectively. Presence of lameness and elevated SAA (≥50 µg/mL) were used as exclusion criteria. Inter-observer reliability was analyzed by intra-class correlation coefficients (ICC). HGS scores between groups were compared using Welch's and Brown Forsythe tests (p < 0.05). Overall, HGS ICC was "excellent" (0.75). No significant differences (p = 0.566) were observed in HGS scores between horses with and without gastric lesions (mean, 95% CI; 3.36, 2.76-3.95 and 3, 1.79-4.20, respectively). HGS was not influenced by the presence or severity of EGUS in this current study. Further studies investigating the use of different pain scales in horses with EGUS are needed.

Risk Factors for Exercise-Associated Sudden Cardiac Death in Thoroughbred Racehorses.

Cardiac arrhythmias resulting in sudden cardiac death (SCD) are an important cause of racehorse fatalities. The objective of this study was to determine risk factors for SCD in Thoroughbreds by evaluating a sample with a policy of mandatory post-mortem following racing or training fatalities. Risk factors were compared between case horses with SCD (n = 57) and control horses with other fatal injury (OFI, n = 188) by univariable and multivariable logistic regression. Survival in years for horses with SCD was compared to OFI using the Kaplan−Meier method with log rank test. The following variables were most important in the multiple logistic model: Horses with SCD were more likely to die during training than during racing, SCD (42/57, 74%) vs. OFI (82/188, 44%; odds ratio [OR], 95% confidence interval [CI], 2.5, 1.2−5.4; p = 0.01), had fewer lifetime starts, median (interquartile range [IQR]), SCD (3.0 [0.0−9.0]) vs. OFI (9.0 [0.0−22.8]; OR, 95% CI, 0.96, 0.9−1.0; p = 0.02 and were less likely to be entire (uncastrated) males, SCD 9/57 (16%) vs. OFI (46/188, 25%; OR, 95% CI, 0.47, 0.1−0.9; p = 0.03). Survival in years (median (IQR)) for horses with SCD was 3.6 (3.1−4.4), which was shorter than OFI (4.5 [3.1−6.0], hazard ratio, 95%CI, 1.6,1.2−2.3; p < 0.001). SCD occurs more commonly in training than racing, which suggests exercise intensity is less important in precipitating this fatality. In this study, SCD occurred early in the careers of affected horses.

The Neuroimmune Interface and Chronic Pain Through the Lens of Production Animals.

Communication between the central nervous system (CNS) and the immune system has gained much attention for its fundamental role in the development of chronic and pathological pain in humans and rodent models. Following peripheral nerve injury, neuroimmune signaling within the CNS plays an important role in the pathophysiological changes in pain sensitivity that lead to chronic pain. In production animals, routine husbandry procedures such as tail docking and castration, often involve some degree of inflammation and peripheral nerve injury and consequently may lead to chronic pain. Our understanding of chronic pain in animals is limited by the difficulty in measuring this pathological pain state. In light of this, we have reviewed the current understanding of chronic pain in production animals. We discuss our ability to measure pain and the implications this has on animal welfare and production outcomes. Further research into the neuroimmune interface in production animals will improve our fundamental understanding of chronic pain and better inform human clinical pain management and animal husbandry practices and interventions.

Validation of an equine fitness tracker: ECG quality and arrhythmia detection.

BACKGROUND: Cardiac arrhythmias in exercising horses are the focus of much interest, both in terms of what is considered normal and potential associations with poor performance and sudden cardiac death. One barrier to performing large-scale studies is the lack of an easily applicable device, to allow recording of large numbers of high-quality exercising electrocardiograms (ECGs). The EquimetreTM is a new wearable device which records a single lead ECG, amongst other parameters. Validation of such wearable devices is essential before further studies are undertaken. OBJECTIVES: To evaluate the quality of ECG using the EquimetreTM and compare arrhythmia detection during exercise with the reference TelevetTM system. STUDY DESIGN: Prospective blinded clinical study. METHODS: Simultaneous ECGs were recorded with both systems in 49 healthy horses during exercise. High intensity exercise (>40km/h) was performed by 29 racehorses, and lower intensity exercise for the remainder of the racehorses and show jumpers. Tracings were excluded if >10% artefact was present (duration of artefact relative to duration of exercise). For included ECGs, the duration of artefact was recorded and compared. ECGs were evaluated using Kubios premium software. Arrhythmia detection (yes/no) and arrhythmia classification (sinus arrhythmia, narrow complex of similar morphology to the sinus complexes or wide complex with a different morphology) were compared using Cohen's Kappa coefficient. RESULTS: Nine TelevetTM ECGs and 3 EquimetreTM ECGs were excluded due to artefact >10%. TelevetTM ECGs included significantly more artefact during exercise than EquimetreTM ECGs (5% vs. 0.25% p<0.001). Arrhythmia analysis was performed on 38 horses' paired ECGs. The Kappa coefficient was excellent for arrhythmia detection (K=0.97) and arrhythmia classification (K=0.93). MAIN LIMITATIONS: Relatively low numbers of horses with arrhythmias (n=21) were included. The ECG recordings only provided one lead, making arrhythmia classification challenging in some cases. CONCLUSIONS: The Equimetre device provides a reliable ECG for arrhythmia detection during exercise. This system may be useful clinically and for future large-scale investigations into the occurrence and significance of exercising arrhythmias.

Oral and Stool Microbiome Coalescence and Its Association With Antibiotic Exposure in Acute Leukemia Patients.

Failure to maintain segregation of oral and gut microbial communities has been linked to several diseases. We sought to characterize oral-fecal microbiome community coalescence, ectopic extension of oral bacteria, clinical variables contributing to this phenomenon, and associated infectious consequences by analyzing the 16S rRNA V4 sequences of longitudinal fecal (n=551) and oral (n=737) samples from 97 patients with acute myeloid leukemia (AML) receiving induction chemotherapy (IC). Clustering observed in permutation based multivariate analysis of variance (PERMANOVA) of Bray-Curtis dissimilarity and PCoA plot of UniFrac distances between intra-patient longitudinal oral-stool sample pairs suggested potential oral-stool microbial community coalescence. Bray-Curtis dissimilarities and UniFrac distances were used to create an objective definition of microbial community coalescence. We determined that only 23 of the 92 patients exhibited oral-stool community coalescence. This was validated through a linear mixed model which determined that patients who experienced coalescence had an increased proportion of shared to unique OTUs between their oral-stool sample pairs over time compared to non-coalesced patients. Evaluation of longitudinal microbial characteristics revealed that patients who experienced coalescence had increased stool abundance of Streptococcus and Stenotrophomonas compared to non-coalesced patients. When treated as a time-varying covariate, each additional day of linezolid (HR 1.15, 95% CI 1.06 - 1.24, P <0.001), meropenem (HR 1.13, 95% CI 1.05 - 1.21, P = 0.001), metronidazole (HR 1.13, 95% CI 1.05 - 1.21, P = 0.001), and cefepime (HR 1.10, 95% CI 1.01 - 1.18, P = 0.021) increased the hazard of oral-stool microbial community coalescence. Levofloxacin receipt was associated with a lower risk of microbiome community coalescence (HR 0.75, 95% CI 0.61 - 0.93, P = 0.009). By the time of neutrophil recovery, the relative abundance of Bacteroidia (P<0.001), Fusobacteria (P=0.012), and Clostridia (P=0.013) in the stool were significantly lower in patients with oral-gut community coalescence. Exhibiting oral-stool community coalescence was associated with the occurrence of infections prior to neutrophil recovery (P=0.002), as well as infections during the 90 days post neutrophil recovery (P=0.027). This work elucidates specific antimicrobial effects on microbial ecology and furthers the understanding of oral/intestinal microbial biogeography and its implications for adverse clinical outcomes.

Incidence, recurrence, and outcome of postrace atrial fibrillation in Thoroughbred horses.

BACKGROUND: Atrial fibrillation (AF) impacts performance and horse and jockey safety. Understanding the outcomes of AF identified postrace will better inform regulatory policy. HYPOTHESIS/OBJECTIVES: To investigate the outcomes after episodes of AF identified postrace and determine whether affected horses are at increased risk of additional episodes compared to the general racing population. ANIMALS: Total of 4684 Thoroughbred racehorses. METHODS: Race records for Thoroughbred horses racing in Hong Kong from 2007 to 2017 were reviewed. Horses that performed below expectation were examined by cardiac auscultation and ECG. Incidence and recurrence of AF were compared between horses with and without a history of AF and between horses with paroxysmal and persistent episodes using Fisher's exact test. RESULTS: There were 96 135 race starts during the study. Atrial fibrillation was identified in 4.9% of horses, with an overall incidence of 2.7 episodes per 1000 starts. The incidence of AF in horses after any previous episode (12.8 per 1000 starts) was higher than for horses with no previous episode (2.4 per 1000 starts; odds ratio [OR], 5.3; 95% confidence interval [CI], 3.8-7.6). Recurrence was seen in 64% of horses previously treated for persistent AF, which was higher than recurrence in horses with paroxysmal AF (23%; OR, 5.9; 95% CI, 1.6-21.2). Median duration between episodes was 343 days (range, 34-1065). CONCLUSIONS AND CLINICAL IMPORTANCE: Thoroughbreds are at increased risk of recurrent AF after both paroxysmal and persistent episodes, but the duration of time between episodes varies widely. These findings support a substantial burden of AF among individual Thoroughbred racehorses.

Exercise-associated rhythm disturbances in poorly performing Thoroughbreds: risk factors and association with racing performance.

BACKGROUND: Exercise-associated cardiac rhythm disturbances are common, but there is a lack of evidence-based criteria on which to distinguish clinically relevant rhythm disturbances from those that are not. OBJECTIVES: To describe and characterise rhythm disturbances during clinical exercise testing; to explore potential risk factors for these rhythm disturbances and to determine whether they influenced future racing. STUDY DESIGN: Retrospective cohort using a convenience sample. METHODS: Medical records were reviewed from two clinical services to identify horses with poor performance and/or respiratory noise with both exercise endoscopy and electrocardiography results. Respiratory and ECG findings recorded by the attending clinicians were described, and for polymorphic ventricular rhythms (n = 12), a consensus team agreed the final rhythm characterisation. Several statistical models analysing risk factors were built and racing records were reviewed to compare horses with and without rhythm disturbance. RESULTS: Of 245 racehorses, 87 (35.5%) had no ectopic/re-entrant rhythms, 110 (44.9%) had isolated premature depolarisations during sinus rhythm and 48 (19.6%) horses had complex tachydysrrythmias. Rhythm disturbances were detected during warm-up in 20 horses (8.2%); during gallop in 61 horses (24.9%) and during recovery in 124 horses (50.6%). Most complex rhythm events occurred during recovery, but there was one horse with a single couplet during gallop and another with a triplet during gallop. Fifteen horses (one with frequent isolated premature depolarisations and 14 complex rhythms) were considered by clinicians to be potentially contributing to poor performance. Treadmill exercise tests, the presence of exercise-associated upper respiratory tract obstructions and National Hunt racehorses were associated with rhythm disturbances. The proportion of horses racing again after diagnosis (82%) was similar in all groups and univariable analysis revealed no significant associations between subsequent racing and the presence of any ectopic/re-entrant rhythm, or the various sub-groups based on phase of exercise in which this was detected. MAIN LIMITATIONS: Reliance on retrospective data collection from medical records with no control group. Exercise ECGs were collected using only 1 or 2 leads. Variables examined as risk factors could be considered to be inter-related and our sub-groups were small. CONCLUSIONS: This study confirms a high prevalence of cardiac rhythm disturbances, including complex ectopic/re-entrant rhythms, in poorly performing racehorses. Detection of rhythm disturbances may vary with exercise test conditions and exercise-associated upper respiratory tract obstructions increase the risk of rhythm disturbances.