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Background Donkeys are routinely vaccinated with protocols developed for horses, yet species‐specific data on their immune responses are limited. Hypothesis/Objectives We hypothesized that donkeys exhibit robust T‐cell‐mediated immunity and regulatory adaptation after vaccination, comparable to horses. Animals Thirty‐six healthy, seronegative donkeys (34 mares, 2 stallions), aged 0.5–23 years (median 8 years), from two farms with similar housing and management conditions. Methods Prospective study. Animals were selected based on clinical health assessment and confirmed seronegativity for tetanus and equine influenza. All received a multivalent vaccine containing tetanus toxoid and equine influenza antigens. Blood samples were collected at baseline, 1 month, and 2 months after vaccination. Flow cytometry assessed CD4+, CD8+, and CD4 + FoxP3+ T cells (primary outcomes), and monocyte subsets and B lymphocytes (PanB/CD21+) with intracellular IL‐10, IL‐17, and Ki67 (secondary outcomes). ANOVA with Bonferroni correction (p < 0.05) was used for statistical analysis. Results CD4+ T cells increased from 25.1% ± 1.4% to 37.3% ± 0.7% at month 1, CD8+ from 20.6% ± 1.5% to 32.2% ± 0.9% at month 2 (p < 0.001). CD4 + FoxP3+ peaked at 11.7% ± 0.6% at month 1 (baseline 6.8% ± 0.8%), then returned to baseline. CD14 + MHCII+ and CD14 + MHCII− monocytes declined; CD14 − MHCII+ increased (p < 0.01). PanB/CD21+ cells decreased from 41.5% ± 1.8% to 29.0% ± 1.0%, with significant reductions in IL‐10+, IL‐17+, and Ki67+ subsets (p < 0.001). Conclusions and Clinical Importance Donkeys exhibit strong T‐cell and regulatory immune responses after vaccination, supporting the clinical relevance of applying equine vaccination protocols to donkeys.

Green coffee extract is currently of great interest to researchers due to its high concentration of chlorogenic acid (CGA) and its potential health benefits. CGA constitutes 6 to 10% of the dry weight of the extract and, due to its anti-inflammatory properties, is a promising natural supplement and agent with therapeutic applications. The purpose of our study was to discover the effects of CGA on peripheral blood mononuclear cell proliferation, and the production of pro- and anti-inflammatory cytokines as well as reactive oxidative species (ROS) in horses during exercise. According to the findings, CGA can affect the proliferation of T helper cells. In addition, at a dose of 50 g/mL, CGA increased the activation of CD4+FoxP3+ and CD8+FoxP3+ regulatory cells. Physical activity decreases ROS production in CD5+ monocytes, but this effect depends on the concentration of CGA, and the effect of exercise on oxidative stress was lower in CD14+ than in CD5+ cells. Regardless of CGA content, CGA significantly increased the release of the anti-inflammatory cytokine IL-10. Moreover, the production of IL-17 was greater in cells treated with 50 g/mL of CGA from beginners compared to the control and advanced groups of horses. Our findings suggest that CGA may have immune-enhancing properties. This opens new avenues of research into the mechanisms of action of CGA and possible applications in prevention and health promotion in sport animals.

In a recent study, in which more than a thousand racehorses were evaluated, three potential blood markers were selected. It was concluded that insulin-like growth factor 1 (IGF1), interleukin-1 receptor antagonist (IL-1ra), and matrix metalloproteinase 2 (MMP2) may enable the early detection of horses at risk of injuries. However, in other studies, it was suggested that cytokine concentrations indicate the athlete’s status better than mRNA expression in blood cells. Thus, the purpose of this study was to evaluate changes in IGF1, MMP-2, and novel markers such as interleukin-13 (IL-13) after exercise in horses at different fitness levels as well as after different intensities of exercise. ELISA tests were performed on thirty-one racehorses [n = 31], who were divided into an inexperienced [beginner] group [n = 20] and an experienced [advanced] group [n = 6]. In addition, differences between race and training session were evaluated to see the influence of different intensities of exercise. Blood samples were taken before and after exercise. The basal IGF1 concentration was lower in an inexperienced group (p < 0.01) as well as IL-13 (p < 0.05) in comparison to the experienced group. There were no differences between pre- and post-exercise samples, changes in multiples or between training, and racing exercises. In conclusion, the basal values of some cytokines may appear to be more beneficial in forecasting horse fitness level.

Horses working with humans for recreational purposes are subjected to a variety of external factors that can have a negative impact on their well-being. There is an urgent need for unequivocal evidence from scientific studies to unify methods of welfare verification of working animals. The testosterone/cortisol ratio has recently been proposed as a marker of the propensity for social aggression as one of the stress reactions. In this study, we analyzed testosterone and cortisol blood concentration and ratio to evaluate the stress susceptibility of horses used for recreational purposes. The blood samples were collected from eleven (n = 11) standardbred horses (age 6–10; geldings–mares = 6:5) during the intense leisure exploitation and after the rest season. The cortisol concentration remained unchanged, whereas, despite the small study population, we observed higher testosterone levels during the horses’ intensive exploitation compared to the resting season (p > 0.09). Thus, the testosterone/cortisol ratio was increased during intensive exploitation. We conclude that recreational horseback riding is not an overly stressful activity for horses; however, it may lead to some behavioral abnormalities connected with high testosterone levels. However, more research is needed.

SLC5A8 is a protein coded by the SLC5A8 gene, and has been proposed as a tumor suppressor and iodide transporter. Its expression is reduced in papillary thyroid carcinoma (PTC), yet the mechanisms underlying this phenomenon are largely unknown. We hypothesized that SLC5A8 expression in PTC is reduced by microRNAs and can be modulated by their inhibition. We used real-time PCR to analyze the expression of SLC5A8 and the microRNAs of interest in a set of 49 PTC/normal tissue pairs. We used an in silico approach to identify microRNAs upregulated in PTC and putatively binding to the SLC5A8 transcript. Luciferase assays were performed to confirm the direct binding of synthetic microRNAs to the 3′UTR of SLC5A8. Subsequently, using mir-expressing plasmids and microRNA sponges, including a microRNA sponge designed to simultaneously inhibit three selected microRNAs, we checked the impact of the modulation of microRNAs on endogenous SLC5A8. Finally, we investigated if modulation of SLC5A8 induces changes in transcriptomes. We confirmed the downregulation of SLC5A8 in PTC. In silico analysis revealed microRNAs potentially targeting SLC5A8. Luciferase assay confirmed direct binding between the 3′UTR of SLC5A8 and miR-181a-5p, miR-182-5p, and miR-494-3p. MiR-181a-5p and miR-182-5p were upregulated in PTC. In HEK293 cell lines, transfection with mir-181a- and mir-182-expressing plasmids decreased endogenous SLC5A8 mRNA, while silencing of miR-181a-5p, miR-182-5p, miR-494-3p, and all three microRNAs simultaneously increased SLC5A8 expression; however, only simultaneous inhibition was able to induce changes visible for SLC5A8 protein. Changes in SLC5A8 expression did not alter the whole transcriptome significantly. This study shows microRNA-dependent regulation of SLC5A8 expression and underlines the potential effectiveness of simultaneous inhibition of a few microRNAs to derepress their common target.

Domestic horses routinely participate in vigorous and various athletic activities. This enables the horse to serve as a model for studying athletic physiology and immunology in other species, including humans. For instance, as a model of physical efforts, such as endurance rides (long-distance running/aerobic exercise) and races (anaerobic exercise), the horse can be useful in evaluating post-exercise response. Currently, there has been significant interest in finding biomarkers, which characterize the advancement of training and adaptation to physical exercise in the horse. The parallels in cellular responses to physical exercises, such as changes in receptor expression and blood cell activity, improve our understanding of the mechanisms involved in the body’s response to intense physical activity. This study focuses on the changes in levels of the pro- and anti-inflammatory cytokines and cellular response in the context of post-exercise immune response. Both the direction of changes in cytokine levels and cellular responses of the body, such as proliferation and expression of surface markers on lymphocytes, monocytes and neutrophils, show cross-functional similarities. This review reveals that horses are robust research models for studying the immune response to physical exercise in human athletes.

Increased training loads in both human and equine athletes are generally implemented to improve their physical performance. These loads are tolerated only within appropriate training periodization with regard to recovery time. Otherwise, training overload causes failure in the systemic adaptation, which at first leads to overreaching, and progressively to overtraining syndrome (OTS). Exercise endocrinology, and anabolic/catabolic balance as an indicator of athlete performance status and OTS has continued to attract attention. In human medicine, changes in testosterone and cortisol levels, as well as the testosterone to cortisol ratio (T/C; anabolic index), are suggested to be sensitive stress markers. However, there is a lack of research investigating these parameters for use in equine sports medicine. The aim of the study was to investigate the differences in testosterone, cortisol, and T/C in response to a single training session in two types of equine sports: endurance and race, together with serum amyloid A (SAA), the main acute phase response indicator of physical effort, and the overall health status in horses. Two groups of horses were enrolled in the study: endurance ( n  = 12) and racehorses ( n  = 32) of different fitness level. Blood samples were obtained before and after the exercise. On average, T increased 2.5 times after the race training in experienced racehorses and dropped in endurance horses regardless the fitness level ( p  < 0.05). In endurance horses, a decrease in T/C occurred after training in inexperienced horses ( p  < 0.05). In racehorses, a T/C decrease occurred in the inexperienced group ( p  < 0.05) and an increase in the experienced ( p  < 0.01). In conclusion, T/C ratio was found to be a potentially reliable indicator of fitness status especially in racing horses. These findings provide insight into the physiological response of the horses to different types of exercise and the potential use of hormone levels as markers of performance and adaptation.

Domestic horses routinely participate in vigorous and various athletic activities. This enables the horse to serve as a model for studying athletic physiology and immunology in other species, including humans. For instance, as a model of physical efforts, such as endurance rides (long-distance running/aerobic exercise) and races (anaerobic exercise), the horse can be useful in evaluating post-exercise response. Currently, there has been significant interest in finding biomarkers, which characterize the advancement of training and adaptation to physical exercise in the horse. The parallels in cellular responses to physical exercises, such as changes in receptor expression and blood cell activity, improve our understanding of the mechanisms involved in the body’s response to intense physical activity. This study focuses on the changes in levels of the pro- and anti-inflammatory cytokines and cellular response in the context of post-exercise immune response. Both the direction of changes in cytokine levels and cellular responses of the body, such as proliferation and expression of surface markers on lymphocytes, monocytes and neutrophils, show cross-functional similarities. This review reveals that horses are robust research models for studying the immune response to physical exercise in human athletes.