Establishing proper policies regarding the recognition and prevention of equine heat stress becomes increasingly important, especially in the face of global warming. To assist this, a detailed view of the variability of equine thermoregulation during field exercise and recovery is essential. 13 endurance horses and 12 trotter horses were equipped with continuous monitoring devices [gastrointestinal (GI) pill, heartrate (HR) monitor, and global positioning system] and monitored under cool weather conditions during four endurance rides over a total of 80 km (40 km loops) and intense trotter track-based exercise over 1,540 m. Recordings included GI temperature (Tc), speed, HR and pre- and post-exercise blood values. A temperature time profile curve of Tc was constructed, and a net area under the curve was calculated using the trapezoidal method. Metabolic heat production and oxygen cost of transport were also calculated in endurance horses. Maximum Tc was compared using an independent samples t-test. Endurance horses (mean speed 14.1 ± 1.7 km h–1) reached mean maximum Tc (39.0 ± 0.4°C; 2 × 40 km in 8 horses) during exercise at 75% of completion of Tc exercise and Tc returned to baseline within 60 min into recovery. However, the mean Tc was still 38.8 ± 0.4°C at a HR of 60 bpm which currently governs “fit to continue” competition decisions. Trotters (40.0 ± 2.9 km h–1) reached a comparable mean max Tc (38.8 ± 0.5°C; 12 horses) always during recovery. In 30% of trotters, Tc was still >39°C at the end of recovery (40 ± 32 min). The study shows that horses are individuals and thermoregulation monitoring should reflect this, no matter what type of exercise is performed. Caution is advised when using HR cut-off values to monitor thermal welfare in horses since we have demonstrated how Tc can peak quite some time after finishing exercise. These findings have implications for training and management of performance horses to safeguard equine welfare and to maximize performance.

Aleurone, a layer of the bran fraction, is deemed to be responsible for the positive health effects associated with the consumption of whole-grain products. Studies on rodents, pigs, and humans report beneficial effects of aleurone in five main areas: the reduction of oxidative stress, immunomodulatory effects, modulation of energy management, digestive health, and the storage of vitamins and minerals. Our study is the first aleurone supplementation study performed in horses. The aim of this study was to investigate the effect of an increase in the dose levels of aleurone on the postprandial glucose-insulin metabolism and the gut microbiome in untrained healthy horses. Seven adult Standardbred horses were supplemented with four different dose levels of aleurone (50, 100, 200, and 400 g/day for 1 week) by using a Latin square model with a 1-week wash out in between doses. On day 7 of each supplementation week, postprandial blood glucose-insulin was measured and fecal samples were collected. 16S ribosomal RNA (rRNA) gene sequencing was performed and QIIME2 software was used for microbiome analysis. Microbial community function was assessed by using the predictive metagenome analysis tool Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) and using the Metacyc database of metabolic pathways. The relative abundancies of a pathway were analyzed by using analysis of composition of microbiomes (ANCOM) in R. There was a significant dose-dependent increase in the postprandial time to peak of glucose (p = 0.030), a significant delay in the time to peak of insulin (p = 0.025), and a significant decrease in both the insulin peak level (p = 0.049) and insulin area under the curve (AUC) (p = 0.019) with increasing dose levels of aleurone, with a consideration of 200 g being the lowest significant dose. Alpha diversity and beta diversity of the fecal microbiome showed no significant changes. Aleurone significantly decreased the relative abundance of the genera Roseburia, Shuttleworthia, Anaerostipes, Faecalibacter, and Succinovibrionaceae. The most pronounced changes in the relative abundance at phyla level were seen in Firmicutes and Verrucomicrobia (downregulation) and Bacteroidetes and Spirochaetes (upregulation). The PICRUSt analysis shows that aleurone induces a downregulation of the degradation of L-glutamate and taurine and an upregulation of the three consecutive pathways of the phospholipid membrane synthesis of the Archaea domain. The results of this study suggest a multimodal effect of aleurone on glucose-insulin metabolism, which is most likely to be caused by its effect on feed texture and subsequent digestive processing; and a synergistic effect of individual aleurone components on the glucose-insulin metabolism and microbiome composition and function.

There is a great need for objective external training load prescription and performance capacity evaluation in equestrian disciplines. Therefore, reliable standardised exercise tests (SETs) are needed. Classic SETs require maximum intensities with associated risks to deduce training loads from pre-described cut-off values. The lactate minimum speed (LMS) test could be a valuable alternative. Our aim was to compare new performance parameters of a modified LMS-test with those of an incremental SET, to assess the effect of training on LMS-test parameters and curve-shape, and to identify the optimal mathematical approach for LMS-curve parameters. Six untrained standardbred mares (3–4 years) performed a SET and LMS-test at the start and end of the 8-week harness training. The SET-protocol contains 5 increments (4 km/h; 3 min/step). The LMS-test started with a 3-min trot at 36–40 km/h [until blood lactate (BL) > 5 mmol/L] followed by 8 incremental steps (2 km/h; 3 min/step). The maximum lactate steady state estimation (MLSS) entailed >10 km run at the LMS and 110% LMS. The GPS, heartrate (Polar®), and blood lactate (BL) were monitored and plotted. Curve-parameters (R core team, 3.6.0) were (SET) VLa1.5/2/4 and (LMS-test) area under the curve (AUC>/<LMS), LMS and Aerobic Window (AW) via angular vs. threshold method. Statistics for comparison: a paired t-test was applied, except for LMS: paired Wilcoxon test; (p < 0.05). The Pearson correlation (r > 0.80), Bland-Altman method, and ordinary least products (OLP) regression analyses were determined for test-correlation and concordance. Training induced a significant increase in VLa1.5/2/4. The width of the AW increased significantly while the AUC</>LMS and LMS decreased post-training (flattening U-curve). The LMS BL steady-state is reached earlier and maintained longer after training. BLmax was significantly lower for LMS vs. SET. The 40° angular method is the optimal approach. The correlation between LMS and VMLSS was significantly better compared to the SET. The VLa4 is unreliable for equine aerobic capacity assessment. The LMS-test allows more reliable individual performance capacity assessment at lower speed and BL compared to SETs. The LMS-test protocol can be further adapted, especially post-training; however, inducing modest hyperlactatemia prior to the incremental LMS-stages and omitting inclusion of a per-test recovery contributes to its robustness. This LMS-test is a promising tool for the development of tailored training programmes based on the AW, respecting animal welfare.

Training-induced follow-up of multiple muscle plasticity parameters in postural stability vs. locomotion muscles provides an integrative physiological view on shifts in the muscular metabolic machinery. It can be expected that not all muscle plasticity parameters show the same expression time profile across muscles. This knowledge is important to underpin results of metabolomic studies. Twelve non-competing Standardbred mares were subjected to standardized harness training. Muscle biopsies were taken on a non-training day before and after 8 weeks. Shifts in muscle fiber type composition and muscle fiber cross-sectional area (CSA) were compared in the m. pectoralis, the m. vastus lateralis, and the m. semitendinosus. In the m. vastus lateralis, which showed most pronounced training-induced plasticity, two additional muscle plasticity parameters (capillarization and mitochondrial density) were assessed. In the m. semitendinosus, additionally the mean minimum Feret's diameter was assessed. There was a significant difference in baseline profiles. The m. semitendinosus contained less type I and more type IIX fibers compatible with the most pronounced anaerobic profile. Though no baseline fiber type-specific and overall mean CSA differences could be detected, there was a clear post-training decrease in fiber type specific CSA, most pronounced for the m. vastus lateralis, and this was accompanied by a clear increase in capillary supply. No shifts in mitochondrial density were detected. The m. semitendinosus showed a decrease in fiber type specific CSA of type IIAX fibers and a decrease of type I fiber Feret's diameter as well as mean minimum Feret's diameter. The training-induced increased capillary supply in conjunction with a significant decrease in muscle fiber CSA suggests that the muscular machinery models itself toward an optimal smaller individual muscle fiber structure to receive and process fuels that can be swiftly delivered by the circulatory system. These results are interesting in view of the recently identified important fuel candidates such as branched-chain amino acids, aromatic amino acids, and gut microbiome-related xenobiotics, which need a rapid gut–muscle gateway to reach these fibers and are less challenging for the mitochondrial system. More research is needed with that respect. Results also show important differences between muscle groups with respect to baseline and training-specific modulation.

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