Evaluation of seasonal influences on adrenocorticotropic hormone response to the thyrotropin-releasing hormone stimulation test and its accuracy for diagnosis of pituitary pars intermedia dysfunction.

Pituitary pars intermedia dysfunction (PPID) is an age-related neurodegenerative disorder, affecting >20 % of older horses. There is a need for improved endocrine tests for early disease detection, and the thyrotropin-releasing hormone (TRH) stimulation test has been recommended for diagnosis of early or mild cases. However, it is currently not recommended for year-round use due to marked seasonal variability. The aims of this cohort study were to evaluate effects of month and season on adrenocorticotropic hormone (ACTH) responses to TRH stimulation and to derive monthly cut-offs for PPID diagnosis. Sixty-three horses were assigned to control (n = 17), subclinical PPID (n = 21) and clinical PPID (n = 25) groups, based on a composite reference standard that combined clinical history and examination findings with endocrine test results. TRH stimulation tests were performed monthly for a 12-month period. Circannual changes were evaluated with one- and two-way repeated-measures analysis of variance and receiver operating characteristic curve analysis was used to derive cut-off values for basal and TRH-stimulated ACTH. TRH-stimulated ACTH concentrations were lowest in February-May and highest in August-October. Specificity of both basal and 30 min post-TRH ACTH was generally higher than sensitivity, and TRH stimulation had improved diagnostic accuracy compared to basal ACTH, although its sensitivity was not significantly greater year-round. TRH stimulation tests yielded considerably more positive results than basal ACTH in the subclinical group, but few additional positive results in clinical PPID cases. There were large differences between cut-offs that maximised sensitivity or specificity for TRH-stimulated ACTH, highlighting the importance of considering clinical presentation alongside test results in diagnostic decision-making.

The lipidome of Thoroughbred racehorses before and after supramaximal exercise.

BACKGROUND: A comprehensive study of the effect of supramaximal exercise in lipid homeostasis of Thoroughbreds provides the basis for future research on the role of lipids on energy metabolism in racehorses. OBJECTIVE: To compare the plasma lipidome of Thoroughbreds before and after supramaximal exercise using an untargeted lipidomics approach. STUDY DESIGN: Pilot experimental study. METHODS: Four Thoroughbred horses were used. The maximal oxygen consumption (VO2 max ) was calculated for each horse. Horses then underwent treadmill exercise at the speed for which the oxygen requirements had been calculated to be 115% VO2 max . Plasma samples were obtained before (T0) and immediately (T1), 15 (T2) and 30 (T3) minutes post-exercise, and evaluated using liquid chromatography/mass spectrometry. Data analysis consisted of principal component analysis and one-way repeated measures analysis of variance. RESULTS: A total of 933 plasma lipids were detected. Supramaximal exercise-induced significant changes in the signal intensity of 13 lipids; all ubiquitous in the organism as major components of biological membranes or energy substrates. MAIN LIMITATIONS: A treadmill was used to replicate track conditions. Also, sample size involved only four horses and the statistical analyses failed to achieve the desired power of 80%. CONCLUSIONS: The findings in this pilot study suggest that supramaximal exercise induces changes in specific plasma lipids in Thoroughbred racehorses. While the biological significance of these findings remains to be determined, these results provide baseline information for future studies in lipidomics applied to equine exercise physiology. Further research is warranted to better understand the role of lipids on energy metabolism in Thoroughbred racehorses.

Rhodococcus equi hyperimmune plasma decreases pneumonia severity after a randomised experimental challenge of neonatal foals.

Since a vaccine is not available against Rhodococcus equi, R equi-specific hyperimmune plasma (HIP) is commonly used, although its efficacy remains controversial. The objective of this study was to evaluate the ability of a commercially available HIP to prevent clinical rhodococcal pneumonia in neonatal foals after experimental challenge.

Validation and evaluation of VapA-specific IgG and IgG subclass enzyme-linked immunosorbent assays (ELISAs) to identify foals with Rhodococcus equi pneumonia.

REASONS FOR PERFORMING STUDY: Rhodococcus equi (Rhodococcus hoagii/Prescottella equi) is a common cause of foal pneumonia, but its diagnosis remains a challenge for equine veterinarians. While the VapA-specific (virulence-associated protein A) immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) has low sensitivity and specificity for detecting pneumonic foals, little is known about VapA-specific IgG subclasses. OBJECTIVES: To evaluate the performance of VapA-specific ELISA for IgG and its subclasses IgGa, IgGb and IgG(T) in the early diagnosis of pneumonia caused by R. equi. STUDY DESIGN: Assay validation followed by assessment of diagnostic performance using archived samples from animals of known status. METHODS: Serum samples from exposed (n = 125) and nonexposed adult horses (n = 10) and from experimentally challenged and naturally infected foals were used for ELISA validation. Post mortem and tissue culture records of the last 24 years from the Institute for Experimental Pathology at the University of Iceland in Keldur, Iceland laboratory were evaluated to confirm the absence of R. equi cases in Iceland. The diagnostic performance of VapA-specific IgG and its subclasses was evaluated using banked serum samples from pneumonic (n = 21) and healthy foals (n = 80). To evaluate each IgG assay, a cut-off value was selected based on receiver operating characteristic curve analysis and used to calculate sensitivity and specificity. The intra- and interassay coefficients of variation were calculated for each ELISA. RESULTS: Using sera from Iceland, where R. equi infection has not been reported, the VapA-specific IgG ELISA differentiated exposed from nonexposed horses. When used to identify infected foals, VapA-specific IgG, IgGa and IgGb had no diagnostic value. In contrast, IgG(T) had high sensitivity and specificity. CONCLUSIONS: Horses from Iceland are not exposed to VapA(+) R. equi and can serve as negative controls. VapA-specific IgG subclasses, with the exception of IgG(T), are poor predictors of disease. Further investigation on the use of IgG(T) as a diagnostic tool in field conditions is needed.

VapA-specific IgG and IgG subclasses responses after natural infection and experimental challenge of foals with Rhodococcus equi.

Rhodococcus equi is a common cause of pneumonia in young foals worldwide and has considerable economic effects on the global equine industry. Despite ongoing efforts, no vaccine is currently available to prevent rhodococaal pneumonia. This is due, in part, to an incomplete understanding of the protective immune response to this bacterium. While antibodies to VapA, a lipoprotein produced by virulent R. equi, are useful in differentiating antibody production in response to pathogenic versus non-pathogenic strains, the significance of the humoral response of foals to this lipoprotein remains poorly defined. The objectives of this study were to evaluate changes in VapA-specific IgG and IgG subclasses after exposure and infection of neonatal foals. Experimental foals included those challenged with R. equi at 1 (n=18), 2 (n=4) and 3 (n=6) weeks of age. Confirmed naturally infected (n=7) and not infected (n=3) foals were also included. All foals were bled 24h after birth and weekly thereafter for a period of 8 weeks. Antibody changes over time were evaluated. Following birth, VapA-specific IgGs significantly (p<0.05) decreased over time in all foals as a result of normal decay of passively transferred antibodies. Both VapA-specific IgGa and IgG(T) significantly increased (p<0.05) after experimental challenge, however, the rise in IgG(T) occurred earlier. Only a significant (p<0.05) increase in VapA-specific IgG(T) over time was seen after natural infection. Whether VapA-specific IgG(T) can be used to differentiate rhodococcal from other pneumonias requires further investigation under field conditions.

Administration of commercial Rhodococcus equi specific hyperimmune plasma results in variable amounts of IgG against pathogenic bacteria in foals.

Rhodococcus equi is the most common cause of pneumonia in young foals. A vaccine is not available and the use of R equi-specific hyperimmune plasma (HIP) is common. Despite its widespread use, the efficacy of HIP in preventing disease remains controversial. The objectives of this study were (1) to evaluate the virulence associate protein A (VapA)-specific IgG and IgG subclasses in commercially available R equi HIP and (2) to evaluate serum VapA-specific IgG and IgG subclasses in foals following administration of commercial R equi HIP. Three different lots from four commercial R equi HIP were sampled. VapA-specific IgG and IgG subclasses were evaluated in all samples using an ELISA. Serum was collected from newborn foals either after commercial R equi HIP was administered (n=97) or not (n=70). Serum was also collected from each mare. Administration of HIP significantly (P<0.001) increased VapA-specific IgGs in recipient foals, however, there was a marked variation in VapA-specific IgGs in foals receiving the same product. VapA-specific IgGs were significantly different (P<0.001) between products and varied between lots, with coefficients of variation ranging from 17 to 123 per cent. These results may explain previously reported disparities in HIP efficacy.

Efficacy of a pectin-lecithin complex for treatment and prevention of gastric ulcers in horses.

The objective of this study was to evaluate the effect of a commercial feed supplement containing pectin-lecithin on squamous mucosa ulceration in horses exposed to an experimental ulceration model. Five mares were treated while five mares were controls for this crossover, blinded study. The mares were fed concentrates and hay and were stabled with a two-hour turn out per day for a period of four weeks. The pectin-lecithin complex was fed for the duration of the study on the treated group. At the end of a four-week period, all mares underwent a seven-day alternating feed deprivation (week 5). The study was repeated again after a four-week washout period. Gastroscopy was performed on days 1, 28 and 35 of the study and was digitally recorded. Independent evaluation of the recordings and scoring of the lesions using the Equine Gastric Ulcer Syndrome (EGUS), severity and number scores were performed by three experienced gastroscopists. The prevalence and severity of squamous ulcers significantly increased after intermittent feed deprivation (P<0.001). No significant effect of the treatment was observed (P>0.05). In this study, the addition of a commercially available pectin-lecithin complex to the feed of horses for five weeks did not prevent or minimise the risk for gastric ulceration of the squamous mucosa.

Use of near-infrared spectroscopy to identify trends in regional cerebral oxygen saturation in horses.

REASONS FOR PERFORMING STUDY: Alterations in cerebral haemodynamics may contribute to perianaesthetic complications in horses. Near-infrared spectroscopy (NIRS) is frequently used intraoperatively in man to provide information regarding cerebral perfusion. OBJECTIVES: To determine whether NIRS can identify trends in regional cerebral oxygen saturation (rSO2) in horses and whether there is a correlation between rSO2 and venous oxygen tensions. METHODS: A cerebral oximeter sensor recorded rSO2 from the dorsal sagittal sinus of 6 healthy horses. Values for rSO2, arterial and venous oxygen and carbon dioxide tensions (PaO2, PvO2, PaCO2 and PvCO2 respectively), along with arteriovenous oxygen saturations (SavO2) were recorded in unsedated (recording period [RP] 1), sedated (RP2) and anaesthetised horses (RP3-5) and during recovery (RP6-8). During anaesthesia, horses were ventilated to achieve states of normo- (RP3), hyper- (RP4) and hypocapnoea (RP5). Data were evaluated descriptively and analysed using linear mixed-effects models and Pearson's correlation coefficient. RESULTS: Overall mean ± s.d. values for rSO2, PaO2, PvO2, PaCO2, SavO2 and mean arterial pressure varied significantly by RP (P<0.001). Significant decreases in rSO2 were identified between RP1 and the post anaesthetic periods (P<0.001). No significant differences in rSO2 values were identified between RP1 and the intra-anaesthesia periods or between RP3, RP4 and RP5. Significant correlations were identified between rSO2 and PaO2 (r = 0.448, P<0.001), rSO2 and PvO2 (r = 0.512, P<0.001) and rSO2 and SavO2 (r = 0.469, P<0.001). CONCLUSIONS: This is the first study to identify trends in rSO2 in horses using NIRS. A positive correlation was identified between rSO2 and PvO2, suggesting that alterations in cerebral oxygenation may be reflected in PvO2 . POTENTIAL RELEVANCE: Near-infrared spectroscopy may be used to monitor trends in rSO2 during equine anaesthesia. Decreasing rSO2 values may act as an early warning signal, alerting clinicians to potential cerebral desaturation events and indicating a need for intervention.

Effects of repeated intra-articular administration of amikacin on serum amyloid A, total protein and nucleated cell count in synovial fluid from healthy horses.

REASONS FOR PERFORMING STUDY: Serum amyloid A (SAA) in synovial fluid has recently been used as a marker for septic arthritis in horses but the effects of repeated intra-articular (IA) administration of amikacin on synovial SAA concentrations are unknown. OBJECTIVES: To report the effect of repeated IA administration of amikacin on SAA, total protein (TP), nucleated cell count (NCC) and differential NCC in synovial fluid of healthy equine joints. METHODS: A controlled, 2 period crossover study was performed on 5 clinically healthy horses. Each intercarpal joint received one of 2 treatments every 48 h for 5 consecutive times: arthrocentesis alone (control group) or arthrocentesis combined with IA administration of 500 mg of amikacin (treatment group). Clinical and lameness examinations were performed daily. Serum SAA and synovial SAA, TP, NCC and differential NCC were measured and statistically compared. Significance level was set at P < 0.05. RESULTS: Horses remained healthy and nonlame throughout the study. Baseline values for all variables were not significantly different between groups. Values for TP in the treatment group were significantly higher than in the control group after the first sample (P < 0.05). In both groups NCC increased significantly (P < 0.05) after the first sample. No significant changes were identified in differential NCC. In both groups, all synovial and most serum SAA concentrations remained below the lower limit of quantification. CONCLUSIONS: Repeated IA administration of amikacin caused increased values of TP and NCC in synovial fluid, with some TP concentrations falling within the range reported for septic arthritis. In contrast, synovial SAA concentrations did not increase in either group. POTENTIAL RELEVANCE: Synovial SAA could serve as a more reliable marker than TP and NCC when evaluating a joint previously sampled or treated with amikacin.