Serum vitamin D concentrations at dry-off and close-up predict increased postpartum urine ketone concentrations in dairy cattle.

Vitamin D is commonly supplemented to dairy cows as vitamin D3 to support calcium homeostasis and in times of low sunlight exposure. Vitamin D has beneficial immunomodulatory and anti-inflammatory properties. Serum 25-hydroxyvitamin D [25(OH)D] concentrations fluctuated during lactation, with the lowest concentrations measured in healthy cows within 7 d of calving. However, it is unknown if serum 25(OH)D concentrations measured during the previous lactation are associated with transition diseases or health risk factors in dairy cattle. We collected serum samples from 279 dairy cattle from 5 commercial dairy herds in Michigan at dry-off, close-up, and 2-10 d in milk (DIM). Vitamin D concentrations were determined by measuring serum 25(OH)D by radioimmunoassay. Total serum calcium was measured by colorimetric methods. Body condition scores (BCS) were assigned at the time of blood collection. Clinical disease incidence was monitored until 30 d postparturition. Separate bivariable logistic regression analyses were used to determine if serum 25(OH)D at dry-off, close-up, and 2-10 DIM was associated with various clinical diseases including mastitis, lameness, and uterine disorders (classified as metritis, retained placenta, or both) and increased urine ketone concentrations at P < 0.05. Among all significant bivariable analyses, multivariable logistic regression analyses were built to adjust for potential confounding variables including parity, BCS, season, and calcium. Receiver operator characteristic (ROC) curve analyses were used to determine optimal concentrations of serum 25(OH)D. We found that higher serum 25(OH)D concentrations at dry-off and close-up predicted increased urine ketone concentrations in early lactation, even after adjustment for confounders. Alternatively, we found that lower serum 25(OH)D at 2-10 DIM was associated with uterine diseases. Optimal concentrations for serum 25(OH)D at dry-off and close-up for lower risk of increased urine ketone concentrations were below 103.4 and 91.1 ng/mL, respectively. The optimal concentration for serum 25(OH)D at 2-10 DIM for uterine diseases was above 71.4 ng/mL. These results indicate that serum 25(OH)D at dry-off and close-up may be a novel predictive biomarker for increased urine ketone concentrations during early lactation. Increased urine ketone concentrations are not necessarily harmful or diagnostic for ketosis but do indicate development of negative energy balance, metabolic stress, and increased risk of early lactation diseases. Predicting that dairy cattle are at increased risk of disease facilitates implementation of intervention strategies that may lower disease incidence. Future studies should confirm these findings and determine the utility of serum 25(OH)D concentrations as a predictive biomarker for clinical and subclinical ketosis.

Reduced serum vitamin D concentrations in healthy early-lactation dairy cattle.

Cattle obtain vitamin D by ingestion or cutaneous exposure to UV light. Dairy cattle diets are frequently supplemented with vitamin D to compensate for limited sun exposure or during times of increased metabolic demands, such as the periparturient period, to maintain calcium homeostasis. Whether housing and supplemental vitamin D practices supply adequate amounts of vitamin D to optimally support the transition from gestation to lactation in dairy cattle is unknown. Our objective was to determine how serum vitamin D concentrations of dairy cows change with season, age, parity, and stage of lactation. Clinically healthy cows (n = 183) from 5 commercial dairies were enrolled in the study. Serum samples were collected at dry off, within 7 d of entering the close-up group, and within 7 d after calving (calving+7). Vitamin D status was determined by measuring serum 25-hydroxyvitamin D [25(OH)D] by radioimmunoassay. We performed repeated-measures mixed-effects linear regression to determine the effects of season, age, parity, and lactation stage (dry off, close-up, and calving+7) on 25(OH)D concentrations in serum. Bivariable analysis indicated that parity, age, and season were not associated with serum 25(OH)D concentrations. Sample period affected 25(OH)D concentrations, with the highest 25(OH)D levels at dry off (99.7 ± 1.9 ng/mL) followed by close up (93.8 ± 2.1 ng/mL), with the lowest levels at calving+7 (82.6 ± 1.7 ng/mL). These data showed a large depletion of 25(OH)D in dairy cattle postpartum compared with late prepartum, although the biological significance of this change in these healthy cattle is unclear. Consumption of serum 25(OH)D by immune system functions and calcium homeostasis in early lactation likely caused the reduction in serum 25(OH)D concentrations after calving. These results suggest that determining whether serum 25(OH)D concentrations are associated with the incidence of transition period disease is an appropriate next step. Assessing the effects of enhanced vitamin D supplementation of cows in early lactation on postpartum diseases may be warranted.

Preliminary safety and biological efficacy studies of ethyl pyruvate in normal mature horses.

REASONS FOR PERFORMING THE STUDY: Endotoxaemia causes substantial morbidity and mortality in horses with colic and sepsis. Ethyl pyruvate is a novel anti-inflammatory medication that improved survival in preclinical models of severe sepsis endotoxaemia and intestinal ischaemia and reperfusion in rodents, swine, sheep and dogs and may be a useful medication in horses. HYPOTHESIS: Ethyl pyruvate has no adverse effects in normal horses and is biologically active based on suppression of proinflammatory gene expression in endotoxin stimulated whole blood, in vitro. METHODS: Physical and neurological examinations, behaviour scores, electrocardiograms and clinicopathological tests were performed on 5 normal healthy horses receiving 4 different doses of ethyl pyruvate. Doses included 0, 50, 100 and 150 mg/kg bwt administered in a randomised crossover design with a 2 week washout period between doses. Biological efficacy was assessed by stimulating whole blood with endotoxin from the horses that received ethyl pyruvate prior to and 1 and 6 h after drug infusion. Gene expression for TNFα, IL-1ß and IL-6 was assessed. RESULTS: There were no effects of drug or dose (0, 50, 100 or 150 mg/kg bwt) on any of the physical or neurological examination, behaviour factors, electrocardiogram or clinical pathological results collected from any of the horses. All parameters measured remained within the normal reference range. There was a significant reduction in TNFα, IL-1ß and IL-6 gene expression in endotoxin stimulated whole blood from horses 6 h after receiving 150 mg/kg bwt ethyl pyruvate. There were no detectable effects on gene expression of any of the other doses of ethyl pyruvate tested. CONCLUSION: We were unable to detect any detrimental effects of ethyl pyruvate administration in normal horses. Ethyl pyruvate significantly decreased proinflammatory gene expression in endotoxin stimulated blood 6 h after drug administration. CLINICAL RELEVANCE: Ethyl pyruvate may be a safe, effective medication in endotoxaemic horses.

Ethyl pyruvate diminishes the endotoxin-induced inflammatory response of bovine mammary endothelial cells.

The endotoxin-induced inflammatory response during coliform mastitis is difficult to control with the currently available therapeutics. Endothelial cells are among the first cell type to be engaged in the inflammatory response and can modulate the severity of inflammation by producing proinflammatory mediators upon endotoxin exposure. Ethyl pyruvate, an ethyl ester of pyruvic acid, can ameliorate endotoxin-induced inflammatory responses by inhibiting the production of proinflammatory mediators in several in vitro and in vivo endotoxemia models. The objective of this study was to determine the effect of ethyl pyruvate on the production of vascular proinflammatory mediators that are associated with the pathogenesis of coliform mastitis. The ability of ethyl pyruvate to reduce the expression of proinflammatory mediators was evaluated in cultured bovine mammary endothelial cells (BMEC) stimulated with endotoxin. Treatment of endotoxin-stimulated BMEC with ethyl pyruvate significantly reduced gene expression of IL-6, IL-8, and intercellular adhesion molecule 1 as well as expression of eicosanoid-producing enzymes, including cyclooxygenase 2 and 15-lipoxygenase 1. This is the first time that the effect of ethyl pyruvate was evaluated in an in vitro BMEC model of coliform mastitis. The ability of ethyl pyruvate to effectively inhibit gene and protein expression of potent vascular proinflammatory mediators in vitro warrants further investigations to assess in vivo efficacy. Ethyl pyruvate is safe for human consumption, and it may be an attractive candidate as a therapeutic in ameliorating the severe pathogenesis associated with coliform mastitis.

Effect of poll flexion and dynamic laryngeal collapse on tracheal pressure in Norwegian Coldblooded Trotter racehorses.

REASON FOR PERFORMING STUDY: Dynamic laryngeal collapse (DLC) associated with poll flexion is a newly diagnosed upper respiratory tract obstructive disorder that causes poor racing performance. OBJECTIVES: To determine if Norwegian Coldblooded Trotters (NCTs) affected with DLC associated with poll flexion differ from normal, elite NCTs based on simple airway mechanics measurements. METHODS: Five normal elite NCTs and 6 NCTs diagnosed previously with DLC underwent treadmill videoendoscopy while tracheal pressures were measured continuously. Alternating head positions were used such that horses were exercised with free head carriage and induced poll flexion at heart rates >200 beats/min. RESULTS: Peak inspiratory tracheal pressures were significantly more negative for horses with DLC compared to the elite horses. This difference was only significant during the exercise phases when the poll region was flexed, P = 0.0015. Head position significantly affected peak inspiratory pressure for both elite and affected horses, P < 0.0001. CONCLUSIONS AND CLINICAL RELEVANCE: Induced poll flexion significantly affected peak inspiratory pressure (PIP) in all horses; however, PIPs were significantly more negative in those affected with DLC. Based upon the tracheal pressure measurements recorded in this study, DLC in NCTs is a severe obstructive upper respiratory tract disorder that is induced by poll flexion.

Incidence of transfusion reactions and retention of procoagulant and anticoagulant factor activities in equine plasma.

BACKGROUND: The extent of preservation of clotting factors and incidence of transfusion reactions to noncommercial equine plasma is not documented. HYPOTHESIS: Equine frozen plasma would retain its coagulation factor activity within the reference range and the incidence of transfusion reactions would be low. ANIMALS: Ten plasma donor horses. Fifty clinically ill hospitalized horses receiving plasma were reviewed to determine the incidence of reactions. METHODS: In vitro study and retrospective case review. Plasma was prepared by gravity sedimentation from whole blood refrigerated for 48 hours. The activities of factors VII through XII, antithrombin (AT), and Protein C were measured. Factor activities were compared for plasma samples obtained before blood collection (S0), after 48 hours of gravity sedimentation at 5 degrees C and after plasma separation (S1), and after 90 days of storage at -20 degrees C (S90). The medical records of 50 consecutive clinically ill horses receiving frozen plasma were reviewed to determine the incidence of transfusion reactions. RESULTS: The combined effect of plasma harvest, gravity sedimentation, decantation, and freezing caused significant reductions in factors IX, (43%P= .0013), X, (33%P= .0001), XI, (48%P= .0008), AT, (10%P= .02), and Protein C (26%P= .0001). Activities for all factors analyzed, except factor X, remained within the reference ranges. Transfusion reactions were recorded for 5/50 horses. CONCLUSIONS AND CLINICAL RELEVANCE: Clotting factors, AT, and Protein C were well preserved. The incidence of reactions to frozen plasma was 10%.

Surgical treatment of colic in the miniature horse: a retrospective study of 57 cases (1993-2006).

REASONS FOR PERFORMING STUDY: It has been suggested that the rate of post operative abdominal adhesions in miniature horses is higher than that for other breeds. However, few reports exist in the veterinary literature describing complications and long-term survival following surgical treatment of colic in these horses. OBJECTIVES: To determine the prevalence of surgical lesions in miniature horses with acute abdominal disease in terms of clinical signs, surgical management, post operative treatment and complications, as well as short- and long-term survival. METHODS: Medical records of 57 American Miniature Horses undergoing surgical treatment for acute abdominal pain at the Michigan State University Large Animal Veterinary Teaching Hospital 1993-2006 were evaluated for clinical information. Owners and trainers were contacted to gain information regarding long-term survival. RESULTS: The most common surgical lesion was a faecalith (38/57 cases) located primarily within the descending colon and most frequently diagnosed in horses age <6 months (19/38 cases). Short-term survival to hospital discharge for horses recovered from anaesthesia was 98% (55/56) with the most common post operative complications being diarrhoea and inappetance. Intra-abdominal adhesions were identified in 2/8 horses requiring a second celiotomy. Long-term follow-up was available for 45 horses and 87% (39/45) were alive at least 12 months after surgery. CONCLUSIONS: As previously reported, faecalith obstruction is a frequent surgical lesion in the miniature horse and is most common in miniature horses age <6 months. The incidence of adhesion formation may be lower than previously reported.

Reproductive careers of Thoroughbred broodmares before and after surgical correction of ≥360 degree large colon volvulus.

BACKGROUND: Limited data exist describing broodmare longevity and reproductive efficiency after surgical correction of ≥360 degree large colon volvulus (° LCV). OBJECTIVES: Compare career duration and foals delivered for broodmares before and after ≥360° LCV surgery. STUDY DESIGN: Retrospective case series. METHODS: Broodmares registered with The Jockey Club that had surgical correction of ≥360° LCV and survived to hospital discharge at Rood and Riddle Equine Hospital from 1 January 2000 to 31 December 2015 were included. Information was collected from the hospital's medical data base and The Jockey Club produce records about the mares' reproductive careers. Data were evaluated using parametric and nonparametric tests, P≤0.05. RESULTS: Mares that were bred but never foaled prior to surgery (n = 19) had shorter careers (mean ± standard deviation [s.d.]), 4.4 ± 4.5 years, and fewer foals, 3.1 ± 3.3, compared with mares that delivered ≥1 foal before surgery (n = 565), 10.4 ± 4.5 years and 7.4 ± 3.4 foals, respectively, P<0.001. Broodmares that delivered foals before surgery produced more foals in the years before surgery, 4.8 ± 3.0, than after surgery, 2.6 ± 2.4, P<0.001, and had longer breeding careers, 5.9 ± 3.8 vs. 4.5 ± 3.3 years before compared with after surgery, P<0.001. No significant differences in career length or number of foals delivered were detected for mares with a single compared with multiple LCV surgeries. Mares that were 3-11 years old at the time of surgery had significantly more foals after surgery compared with mares ≥12 years old, P<0.001, as expected. MAIN LIMITATIONS: Retrospective collection of data. CONCLUSIONS: Broodmares had productive careers following surgery for ≥360° LCV that were largely influenced by the mares' age at the time of surgery.

Electromyographic activity of the palatinus and palatopharyngeus muscles in exercising horses.

REASONS FOR PERFORMING STUDY: Determining the respiratory related activity of the palatinus and palatopharyngeus muscles in exercising horses is relevant because dysfunction of these muscles has been implicated in the pathogenesis of dorsal displacement of the soft palate. OBJECTIVE: To determine if the palatinus and palatopharyngeus muscles have respiratory activity that increases with intensity in exercising horses. METHODS: Electromyographic activity was measured in the palatinus and palatopharyngeus muscles using bipolar fine-wire electrodes while the horses completed an incremental exercise treadmill protocol. RESULTS: Both muscles displayed synchronous expiratory activity that increased significantly (P < 0.05) with exercise intensity. Phasic expiratory activity of the palatinus increased 390 +/- 98%, whereas phasic expiratory activity of the palatopharyngeus increased by 198 +/- 30% as the treadmill speed increased from 6 to 12 m/s. CONCLUSIONS: The palatinus and palatopharyngeus muscles may be important respiratory muscles, functioning to stabilise the position of the soft palate during intense exercise. CLINICAL RELEVANCE: The predominant expiratory activity of these muscles may be associated with specific muscle function related to exercise or distinct upper airway phenomena of an obligate nasal breather, such as the horse.

The effect of uterine torsion on mare and foal survival: a retrospective study, 1985--2005.

REASONS FOR PERFORMING STUDY: Anecdotal speculation suggests that prognosis for survival of mares and foals following correction of uterine torsion has improved over the past 30 years. OBJECTIVES: To determine statistically the outcome of uterine torsion according to duration of clinical signs, stage of gestation, parity, physical examination findings, method of correction, prognosis for survival and reproductive health of the mare, and prospects for the foal within the neonatal period. METHODS: This retrospective study combined cases from 4 equine referral hospitals. RESULTS: The stage of gestation at which uterine torsion occurred was a risk factor for survival of mare and foal. Overall mare survival was 53/63 (84%); when uterine torsion occurred at < 320 days gestation, 36/37 (97%) of mares survived compared to 17/26 (65%) survival rate when uterine torsion occurred at > or = 320 days gestation. Overall foal survival was 54% (29/54). When uterine torsion occurred at < 320 days gestation, 21/29 (72%) foals survived compared to 8/25 (32%) when uterine torsion occurred at > or = 320 days gestation. Thirty mares were discharged from the hospital carrying a viable fetus following uterine torsion correction and 25/30 (83%) of these mares delivered live foals that survived beyond the neonatal period. CONCLUSIONS: Prognosis for survival for mares and foals following uterine torsion is good and improves if torsion occurs < 320 days compared to > or = 320 days gestation. CLINICAL RELEVANCE: Gestational timing of uterine torsion should be considered when advising clients about the prognosis for survival of the mare and foal. The prognosis for a mare delivering a live foal is good if the mare is discharged from the hospital following uterine torsion correction with a viable fetus.

Airway inflammation in Michigan pleasure horses: prevalence and risk factors.

REASONS FOR PERFORMING STUDY: Although subclinical airway inflammation is thought to be common in horses, there is little information on its prevalence and none on risk factors. OBJECTIVE: To determine the prevalence and risk factors for an increased number of inflammatory cells and for mucus accumulation in the trachea of pleasure horses. METHODS: Horses (n = 266) in stables (n = 21) in Michigan were examined endoscopically, once in winter and once in summer 2004. Visible tracheal mucoid secretions were graded 0-5 and inflammatory cell numbers counted in a tracheal lavage sample. Information collected about each horse included age, gender, presence of cough, percent time indoors and source of roughage. The repeated measures were analysed by generalised estimating equations and linear mixed models. RESULTS: Horses eating hay, especially from round bales, had the most neutrophils, whereas horses feeding from pasture had the fewest. Being female and being outdoors in winter were associated with increased numbers of inflammatory cells. Older horses had fewer macrophages than young horses. More than 70% of horses had >20% neutrophils in tracheal lavage. Twenty percent of horses had a mucus accumulation score >1; 17% had both a mucus score >1 and >20% neutrophils. The significant risk factors for mucus accumulation >1 were age >15 years, feeding on hay as compared to pasture, and being outdoors for more than 80% time in winter. Even though mucus accumulation score >1 was a risk factor for cough, only half of such horses coughed. Cough and mucus accumulation were associated with increased number of neutrophils. CONCLUSIONS: In comparison to pasture feeding, hay feeding, particularly from round bales, was associated with an increased number of neutrophils in the airway. Being outdoors in winter was associated with increased numbers of inflammatory cells and with mucus accumulation. Because 70% of horses have >20% neutrophils, this value should not be used as the sole indicator of airway inflammation. POTENTIAL RELEVANCE: The study reinforces the importance of hay feeding and older age as risk factors for inflammatory airway disease. Horses that do not have 'heaves' may be best kept indoors when winters are cold.

Effect of tracheal mucus and tracheal cytology on racing performance in Thoroughbred racehorses.

REASON FOR PERFORMING STUDY: Accumulations of mucus within the trachea are often found during endoscopic examinations of the airways of poorly performing racehorses, but the clinical importance of this finding is unknown. OBJECTIVES: To determine the effect of tracheal mucus, pharyngeal lymphoid hyperplasia (PLH) and cytological indices of tracheal aspirate on racing performance in Thoroughbred horses assessed by race place and whether the horse was raced. METHODS: Endoscopic examination of the nasopharynx, larynx and trachea was performed, and a tracheal aspirate obtained monthly at Thistledown racetrack from April to December, 2002 and 2003. Horses received a score of 0-4 for the degree of PLH and 0-4 for the amount of mucus visible in the trachea. The tracheal aspirate was assessed for turbidity, and total and differential cell counts. Generalised estimating equations models were used as repeated measures models for each risk factor and the level of association assessed through the risk factor's P value in the model. RESULTS: Moderate to severe tracheal mucus (2-4) was a risk factor for poor racing performance. There was no association between degree of PLH, cell counts or turbidity of tracheal wash fluid and racing performance. However, horses that raced had higher total neutrophil counts in tracheal wash aspirates than horses that did not race. CONCLUSIONS: Grades 2-4 tracheal mucus should be considered a potential cause of poor racing performance in Thoroughbred horses. CLINICAL RELEVANCE: Because moderate to severe tracheal mucus accumulation, and not increased tracheal neutrophils, was a risk factor for poor racing performance, functionally significant airway inflammation may best be confirmed by the presence of mucus rather than increased number of neutrophils in the trachea.

Increased serum nonesterified fatty acid and low ionised calcium concentrations are associated with post partum colic in mares.

REASONS FOR PERFORMING STUDY: Increased serum nonesterified fatty acids (NEFA) and decreased serum electrolytes are linked to abdomasal displacements in post partum dairy cattle. Post partum colic in mares may be associated with metabolic changes specific to pregnancy and the periparturient period. OBJECTIVES: To determine if fluctuations in serum NEFA, ionised calcium (iCa) and magnesium (iMg) occurred in periparturient mares and if these alterations were associated with post partum colic. STUDY DESIGN: Longitudinal observational study. METHODS: Mares from 3 farms in central Kentucky were enrolled. Blood samples were collected 14 days prior to the estimated foaling date, within 4 days post parturition, and 14 and 28 days after foaling for batch analysis of serum NEFA, iCa and iMg. Health information was provided by farm managers and veterinarians. Data were analysed using Kruskal-Wallis χ(2) statistic for nonparametric data and a matched case/control approach. Repeated measures logistic regression models were developed. RESULTS: Serum NEFAs were higher at 14-1 day before foaling (mean ± s.d., mmol/l), 0.28 ± 0.12, P = 0.04 and from foaling to 4 days after foaling, 0.29 ± 0.20 (P = 0.05) in mares that developed colic compared with those that did not colic, 0.19 ± 0.05 and 0.21 ± 0.14, respectively. Ionised calcium was lower at 15-28 days post foaling in mares that showed colic, 1.50 ± 0.17 compared to mares that did not colic, 1.60 ± 0.12, P = 0.02. Risk of colic in post partum mares increased 38% for each 0.1 mmol/l increase in serum NEFA (odds ratio = 1.38, 95% confidence interval 1.06-1.81, P = 0.02). CONCLUSIONS: Mares with post partum colic had significantly higher serum NEFA and lower iCa prior to the colic episode compared with mares that did not develop colic. Monitoring these metabolic alterations may lead to predictive and preventive colic strategies for post partum mares.

Laryngeal advancement surgery improves swallowing function in a reversible equine dysphagia model.

REASONS FOR PERFORMING STUDY: Pharyngeal dysphagia is a debilitating, sometimes fatal condition in horses, with multiple aetiologies. The pathophysiology is complex and not fully understood. Treatment is largely supportive. Laryngeal advancement surgery may diminish symptoms of dysphagia and improve swallowing in affected horses. OBJECTIVES: 1) to induce reversible moderate and marked pharyngeal dysphagia by regional anaesthesia of branches of the glossopharyngeal (IX), vagus (X) and hypoglossal (XII) nerves; 2) to characterise the dysphagia produced by each model; and 3) to determine whether laryngeal advancement surgery improves swallowing in these models. STUDY DESIGN: Experimental design using 6 adult horses. METHODS: Two dysphagia models were produced by blocking IX, the pharyngeal branch of X and XII unilaterally (moderate model) and only the pharyngeal branch of X bilaterally (marked model) within the guttural pouches. Both models were performed on each horse before and after surgery in order to assess the effectiveness of the surgical procedure as a potential treatment for pharyngeal dysphagia. Dysphagia was scored by partly blinded observers on a scale of 0-12 based on observations of eating (nonblinded), endoscopic examinations and fluoroscopic swallowing (blinded), where 0 = normal swallow and 12 = severe dysphagia with tracheal aspiration. Data were analysed by 3-factor ANOVA, with significance taken as P<0.05. RESULTS: Dysphagia models were reversible, and horses swallowed normally within 3 h of model induction. The marked dysphagia model impaired movement of feed from the base of the tongue to the oesophagus and caused severe airway contamination. The average dysphagia score (mean ± s.d.) for the marked dysphagia model was 10.6 ± 1.1 before surgery and 6.1 ± 4.3 after surgery (P = 0.007). Laryngeal advancement surgery did not significantly improve the dysphagia scores in the moderate model (P = 0.5). CONCLUSIONS: Laryngeal advancement surgery may improve swallowing and reduce aspiration in horses affected with diseases that cause pharyngeal dysphagia.

Electromyographic activity of the stylopharyngeus muscle in exercising horses.

REASONS FOR PERFORMING STUDY: There is a need to understand the process which leads to failure of recruitment of the stylopharyngeus muscle in clinical cases of nasopharygeal collapse. We therefore studied the timing and intensity of stylopharyngeus muscle activity during exercise in horses. OBJECTIVE: To measure the electromyographic (EMG) activity of the stylopharyngeus muscle in exercising horses and correlate it with the breathing pattern. METHODS: Five horses were equipped with a bipolar fine wire electrode placed on the stylopharyngeus muscle and a pharyngeal catheter. The horses exercised on a treadmill at speeds corresponding to 50 (HRmax50), 75 and 100% of maximum heart rate, and EMG activity of the stylopharyngeus muscle and upper airway pressures were recorded. The EMG activity of the stylopharyngeus muscle was then correlated to the breathing pattern and the activity quantified and reported as a percentage of the baseline activity measured at HRmax50. RESULTS: There was ongoing activity of the stylopharyngeus muscle throughout the breathing cycle; however, activity increased towards the end of expiration and peaked early during inspiration. Tonic activity was present during expiration. Peak, mean electrical and tonic EMG activity increased significantly (P<0.05) with exercise intensity. CONCLUSION: The stylopharyngeus muscle has inspiratory-related activity and tonic activity that increases with speed. POTENTIAL RELEVANCE: The stylopharyngeus muscle is one of a group of upper airway muscles that function to support and maintain the patency of the nasopharynx during inspiration. Failure of recruitment of the stylopharyngeus muscle during exercise is a potential explanation for clinical cases of dorsal pharyngeal collapse, but further work investigating the activity of the stylopharyngeus muscle in horses affected by this disease is needed.

The effect of bilateral glossopharyngeal nerve anaesthesia on swallowing in horses.

REASONS FOR PERFORMING STUDY: Dysfunction of the glossopharyngeal nerve has been implicated as a cause of dysphagia in horses. However, recent studies have indicated that this is not the case. OBJECTIVES: To determine whether bilateral glossopharyngeal nerve anaesthesia would cause dysphagia in horses or result in measurable alterations in the timing, function, or sequence of swallowing. METHODS: Swallowing was evaluated in 6 normal horses with and without bilateral glossopharyngeal nerve anaesthesia. Swallowing dynamics were assessed subjectively and objectively based on time from prehension of food until swallowing, number of tongue movements until initiation of swallowing, depth of bolus at the base of the tongue prior to initiation of swallow and evidence of tracheal aspiration using fluoroscopy and endoscopy. RESULTS: There was no evidence of aspiration or dysphagia in horses before or after bilateral glossopharyngeal nerve block. No observed or measured differences in swallowing sequence or function could be detected in blocked compared to unblocked horses. However, there was a trend in blocked horses for the number of tongue pushes and the time to swallowing to be increased. CONCLUSIONS: Glossopharyngeal nerve function may not be essential for normal swallowing function in otherwise healthy horses. POTENTIAL RELEVANCE: Clinically, normal swallowing is not an appropriate test of glossopharyngeal nerve function and dysphagic horses should not be assumed to have glossopharyngeal nerve dysfunction.

Changes in the faecal microbiota of mares precede the development of post partum colic.

REASONS FOR PERFORMING STUDY: Disruptions in the gastrointestinal microbiota may trigger development of post partum colic. OBJECTIVES: To determine the effects of the periparturient period on the faecal microbiome and identify associations between the faecal microbiota and post partum colic. STUDY DESIGN: Longitudinal case-control study. METHODS: Pre- and post partum faecal samples were collected from mares on 3 farms in central Kentucky. Next generation sequencing of the V4 region of the 16S rRNA gene was performed on samples from 13 mares that developed colic, 13 mares that did not display colic and 5 nonpregnant controls. RESULTS: There were 4,523,727 sequences from 85 samples evaluated (mean ± s.d. 53,220 ± 29,160, range 8442-122,535). Twenty-five phyla were identified, although only Firmicutes, Verrucomicrobia, Actinobacteria and Proteobacteria were present at a relative abundance of 1% or greater. The faecal microbiota of late-term mares differed from nonpregnant mares, with differences in microbial community membership and structure but not the relative abundance of major phyla. There was limited impact of foaling and the post partum period on the faecal microbiome. Faecal samples obtained from mares prior to episodes of colic had significantly higher relative abundance of Proteobacteria (8.2%, P = 0.0006) compared with samples from mares that did not display colic (3.7%). All samples with a relative abundance of Firmicutes of ≤50% preceded colic, as did 6/7 (86%) samples with >4% Proteobacteria. Differences in microbiota membership and structure were also present between mares that developed large colon volvulus and matched controls that did not have colic. Sixty-one indicator operational taxon units were identified for the control (vs. volvulus) samples, and these were dominated by Lachnospiraceae (n = 38) and Ruminococcaceae (n = 8). CONCLUSIONS: Foaling had minimal effects on the mares' faecal microbiota. Numerous differences in the faecal microbiota preceded colic. Associations between Firmicutes (particularly Lachnospiraceae and Ruminococcaceae) and Proteobacteria and development of colic could lead to measures to predict and prevent colic. The Summary is available in Chinese - see Supporting information.

Effect of sternothyrohyoid myectomy on upper airway mechanics in normal horses.

The effect of transection of the sternothyroideus and sternohyoideus muscles on upper airway mechanics was investigated in exercising horses. Upper airway mechanics of six Standardbred horses were measured at rest and during exercise before and 24 h and 2 wk after sternothyrohyoid myectomy. Transnasal tracheal and pharyngeal catheters connected to differential pressure transducers were used to measure tracheal and pharyngeal pressures. A pneumotachograph mounted on the rostral end of an airtight face mask was used to measure airflow. Horses ran at 50, 75, and 100% of maximal O2 consumption on a treadmill. Twenty-four hours after sternothyrohyoid myectomy, no significant difference was detected in tracheal, pharyngeal, or translaryngeal inspiratory and expiratory pressures and impedances, inspiratory and expiratory flows, and respiratory frequency. Two weeks after sternothyrohyoid myectomy, there was a statistically significant increase in translaryngeal inspiratory pressure (P = 0.035) and tracheal inspiratory pressure (P = 0.032) compared with preoperative measurements. Two weeks after sternothyrohyoid myectomy, there was a statistically significant increase in translaryngeal inspiratory resistance (P = 0.017) and tracheal inspiratory resistance (P = 0.023) compared with preoperative values. Increased translaryngeal and tracheal inspiratory pressures and resistances after sternothyrohyoid myectomy suggest that the sternothyroideus and sternohyoideus muscles act to increase or maintain upper airway patency and stability in normal horses.

Effects of stylopharyngeus muscle dysfunction on the nasopharynx in exercising horses.

REASONS FOR PERFORMING STUDY: Nasopharyngeal collapse has been observed in horses as a potential cause of exercise intolerance and upper respiratory noise. No treatment is currently available and affected horses are often retired from performance. OBJECTIVE: To determine the effect of bilateral glossopharyngeal nerve block and stylopharyngeus muscle dysfunction on nasopharyngeal function and airway pressures in exercising horses. METHODS: Endoscopic examinations were performed on horses at rest and while running on a treadmill at speeds corresponding to HRmax50, HRmax75 and HRmax, with upper airway pressures measured with and without bilateral glossopharyngeal nerve block. RESULTS: Bilateral glossopharyngeal nerve block caused stylopharyngeus muscle dysfunction and dorsal nasopharyngeal collapse in all horses. Peak inspiratory upper airway pressure was significantly (P = 0.0069) more negative at all speeds and respiratory frequency was lower (P = 0.017) in horses with bilateral glossopharyngeal nerve block and stylopharyngeus muscle dysfunction compared to control values. CONCLUSIONS: Bilateral glossopharyngeal nerve anaesthesia produced stylopharyngeus muscle dysfunction, dorsal pharyngeal collapse and airway obstruction in all horses. POTENTIAL RELEVANCE: The stylopharyngeus muscle is probably an important nasopharyngeal dilating muscle in horses and dysfunction of this muscle may be implicated in clinical cases of dorsal nasopharyngeal collapse. Before this information can be clinically useful, further research on the possible aetiology of stylopharyngeus dysfunction and dysfunction of other muscles that dilate the dorsal and lateral walls of the nasopharynx in horses is needed.

Stabling is associated with airway inflammation in young Arabian horses.

We examined the effect of stabling on upper and lower airway inflammation in 14 yearling Arabian horses that had been at pasture since birth. Horses were divided into 2 groups of 7. One group was stabled for 3 months and the other remained at pasture. The groups were then switched over for another 3 months. The nasopharynx, guttural pouches and trachea were examined endoscopically and bronchoalveolar lavage performed every month. An upper airway inflammation score was devised based on the magnitude of pharyngeal lymphoid hyperplasia and guttural pouch inflammation. During stabling this score remained constant, whereas it decreased during the 3 months at pasture. Stabling was also associated with a higher number and percentage of neutrophils in bronchoalveolar lavage fluid and with a smaller percentage of lymphocytes. There was no correlation between upper airway inflammation score and bronchoalveolar lavage cytology. During a nasal occlusion test, dorsal displacement of the soft palate occurred more times in stabled than in pastured horses, but this was heavily biased by the results from one animal. We conclude that stabling is associated with inflammation of both the upper and lower airway of young horses.