Plasma relaxin, estrogens and progestogens concentration in coldblooded and hot-blood mares in the second half of gestation.

Monitoring estrogens, progestogens, and relaxin plasma concentration can be important for determining pregnancies at risk of failure in mares. However, significant breed-related differences in values of reproductive hormones are known, and data concerning coldblooded (CB) mares are missing. The aim of the study was to determine and compare serum relaxin, estrone sulphate (E1S), estradiol-17ß (E2), and progestogens concentration in pregnant CB and hot-blood (HB) mares during the second half of gestation. Blood samples obtained from 94 healthy mares in the second half of gestation with known mating dates were analyzed. Samples were collected from 36 CB mares aged from 3 to 16 years and 58 HB mares aged from 4 to 17 years. The hormone concentrations were measured using immunoenzymatic equine species-specific commercial kits. The serum relaxin concentration was almost three times lower in CB, whereas E1S and E2 almost twice as low, as compared to HB mares (P < 0.05). In conclusion, the extremely low concentration of relaxin in CB mares could be one reason for the increased amounts of difficult parturition requiring veterinary intervention noted in these breeds.

The Influence of Selected Factors on the Nutritional Value of the Milk of Cold-Blooded Mares: The Example of the Sokólski Breed.

This study assessed the effect of access to pasture, lactation number, and foals' sex on the nutritional value of milk (79 samples) from nine mares. The following were analysed: content of dry matter, protein, fat, lactose, and ash; percentages of α-lactalbumin (α-La), ß-lactoglobulin (ß-Lg), serum albumin (SA), immunoglobulins (Ig), lactoferrin (Lf), and lysozyme (Lz) in the total protein; and the fatty acid profile. Mares without access to pastures were shown to produce milk with a higher dry matter content, including fat, lactose, and ash; higher percentages of ß-Lg, α-La, Ig, and Lf; and a better fatty acid profile. The milk from mares with access to pasture contained more protein, including higher percentages of SA and Lz. Milk from mares in lactations 4-6 had the highest fat and protein concentrations and the lowest lactose concentration. The α-La level was highest in lactation 1, Lf in lactations 2-3, and Lz in lactations 4-6. Milk from mares in lactations 4-6 had the best fatty acid profile (the lowest concentration of saturated fatty acids (SFAs) and the highest concentration of monounsaturated fatty acids(MUFA) and polyunsaturated fatty acids (PUFA)). Milk from mothers of female offspring had higher dry matter, fat, and protein concentrations, a higher share of lysozyme, and a better fatty acid profile.

Variation of Physiological and Behavioural Parameters during the Oestrous Cycle in Mares.

The behaviour of oestrous mares is well-known in horse breeding. However, alterations in their physiological and behavioural indices during the whole oestrous cycle are scarcely known. The objective of the study was to analyse changes in cardiac activity variables, rectal and superficial temperatures, behaviour towards humans and conspecifics, and the time of standing and locomotor activity in mares during their oestrous cycle. Fifteen adult mares in oestrus were examined in the morning and evening (six successive days) and in dioestrus (five days-once every third day). The oscillation of physiological and behavioural variables accompanies changes in mares' sexual behaviour. Most physiological variables studied in oestrus indicate the elevated activity of the adrenergic nervous system and, opposite to that, both behaviour towards humans and conspecifics and the time of standing relate to a relaxed state. The end of oestrus, manifested by a rapid decrease in most of the physiological variables studied, is followed by changes of behavioural variables at the beginning of dioestrus. The time of locomotor activity arises at the end of oestrus. The outcomes may contribute to the knowledge of, among others, mare owners who evaluate the oestrus by mares' sexual behaviours without regarding other rhythmically changing variables.

Can Sound Alone Act as a Virtual Barrier for Horses? A Preliminary Study.

Virtual fencing is an innovative alternative to conventional fences. Different systems have been studied, including electric-impulse-free systems. We tested the potential of self-applied acoustic stimulus in deterring the horses from further movement. Thirty warmblood horses were individually introduced to a designated corridor leading toward a food reward (variant F) or a familiar horse (variant S). As the subject reached a distance of 30, 15 or 5 m from a finish line, an acute alarming sound was played. Generally, a sudden and unknown sound was perceived by horses as a threat causing an increase in vigilance and sympathetic activation. Horses' behaviour and barrier effectiveness (80% for F vs. 20% for S) depended on motivator (F/S), while the cardiac response indicating some level of stress was similar. The motivation for social interactions was too strong to stop the horses from crossing a designated boundary. Conversely, the sound exposure distance did not vary the barrier effectiveness, but it differentiated HRV responses, with the strongest sympathetic activation noted at a distance of 5 m. Thus, the moment of a sound playback has important welfare implications. Due to the limited potential of sound as a virtual barrier, auditory cues cannot be used as an alternative for conventional fencing.

Minimizing the Effects of Social Isolation of Horses by Contact with Animals of a Different Species: The Domestic Goat as an Example.

This study aimed to perform a comparative analysis of the horses' heart rate parameters and locomotor activity in a herd or isolation, with or without the company of goats. Twenty horses were tested in a paddock, accompanied (or not) by three goats. The experiment comprised four tests (a control test of a herd of horses without goats, a horse isolation test without goats, a test of a herd of horses with goats and a test of an isolated horse with goats). The horse's locomotor behavior, and the HR, RR, rMSSD, LF, HF, and LF/HF were recorded. The data analysis included a 15-min rest, procedural and recovery HR/HRV periods, and a 5-min period at the beginning of the test. The duration of the horses standing in the company of goats increased significantly. The rMSSD parameter was the significantly lowest in the test of a herd of horses with goats. The company of goats in a paddock does not eliminate the emotional effects of the phenomenon. However, the locomotor behavior decreases. Goats in a paddock can provide a positive distraction for horses in a herd as a decrease in emotional excitability can be regarded as having a relaxing impact on a different animal species.

Correction: Wisniewska et al. Heterospecific Fear and Avoidance Behaviour in Domestic Horses (Equus caballus). Animals 2021, 11, 3081.

The authors wish to make the following correction to this paper [...].

Horses' Response to a Novel Diet: Different Herbs Added to Dry, Wet or Wet-Sweetened Oats.

The commercial horse feed industry uses palatants to mask undesirable tastes of feeds and enhance product acceptance. However, an unknown odour or taste may also hinder feed intake, due to, inter alia, novelty. The objective of the study was to assess the horses' response to novel diet: five different herbs added alternately to dry, wet or wet-sweetened oats. Twenty adult horses were given different diet combinations of a feed presentation and a herb: field mint, common yarrow, common chamomile, common sage and common nettle, consecutively, once daily. The response to novelty was assessed regarding traits showing the willingness to consume: times of olfaction and consumption, times and numbers of intervals in consumption and drinking water, and the mass of leftovers. The results show that properties of the herbs studied did not hinder the consumption and only the odour of the dry common sage delayed the intake. Wetting or wetting and sweetening the diet accelerated the intake. In conclusion, herbs in small amounts do not significantly affect the willingness to consume feed. Although wet and wet-sweetened diet presentations may be novel to horses, they increase the feed palatability and can be suggested for use when preparing horse diets.

Behavioural responses of Konik Polski horses to natural, familiar sound of thunderstorm, and unfamiliar similar-sounding sounds of volcanic eruption and sea storms.

BACKGROUND: It is not clear, if modern Konik Polski horses have retained the ability to identify sounds in terms of danger. The aim of the study was to identify differences in their behaviour in response to the reproduction of volcanic eruption and sea storm sounds, assumed to be unfamiliar to these horses, as compared to their response to a thunderclap sound, considered by the horses as potentially dangerous. The study included 13 adult mares of the Konik Polski breed, kept under a free-range system. Their behavioural responses to the reproduction of the three natural sounds with an intensity of over 50 dB, were registered. They were analysed distance of each horse to the central point of the pasture and to the exit from the enclosure, and time and/or frequencies of elements of behaviour categorised as: increased anxiety (walking, trotting and cantering), vigilance (snoring, vocalisation, high head position, high tail position, sticking together), foraging (time of grazing), comfort (playing, examining the surroundings, sniffing), maintenance of hygiene (rubbing against objects, auto- or allogrooming, rolling) and resting. The obtained data were analysed by the Dwass, Steel and Critchlow-Fligner method using the SAS program. RESULTS: Most of analysed elements increased in response to reproduced sounds and decreased after sounds were stop playing (p < 0.05), however, they were no significant differences in general response to each studied sound. CONCLUSIONS: The responses of horses to similar sounds of both known and unknown origins, i.e. the sound of a thunderstorm, sea storm and volcanic eruption, are similar. The sound stimuli applied were not too stressful for the horses.

Effect of Different Water Cooling Treatments on Changes in Rectal and Surface Body Temperature in Leisure Horses after Medium-Intensity Effort.

Cooling a horse after intensive exercise under hot conditions is commonly recommended. The study aimed to analyze changes in the rectal and surface temperature of the horses subjected to various water cooling treatments. This followed medium-intensity exercise performed by leisure horses under moderate air temperature. The experiment involved a control group without water application, and three variants of water cooling applied to 19 warmblood geldings after medium-intensity effort. Cooling of lower, upper, and lower and upper body parts was performed. In each variant, the rectal and body surface temperatures were measured five times: before; immediately after; and 10, 20, and 30 min after effort. Using water cooling under the studied conditions did not influence a post-exercise decrease in the rectal temperature. The decrease in body surface temperature depended on the used variant of cooling down the horse. Cooling the limbs by pouring water several times changed the surface body temperature from 34.2 ± 0.37 °C to 32.0 ± 0.32 °C and was more efficient than the repeated application of cool water on both the upper and lower body parts, leading to a temperature change from 34.6 ± 0.26 °C to 33.2 ± 0.36 °C. Thus, the application of cold water on the limbs only is sufficient for cooling the horse after medium-intensity exercise under moderate air temperature (about 24 °C).

Heterospecific Fear and Avoidance Behaviour in Domestic Horses (Equus caballus).

Ridden horses have been reported to be fearful of cows. We tested whether cows could provoke behavioural and cardiac fear responses in horses, and whether these responses differ in magnitude to those shown to other potential dangers. Twenty horses were exposed to cow, a mobile object or no object. The time spent at different distances from the stimulus was measured. In a separate test, heart rate (HR), root mean square of successive differences between heartbeats (RMSSD) and the horses' perceived fear were assessed at various distances from the stimuli. The horses avoided the area nearest to all stimuli. During hand-leading, the cow elicited the highest HR and lowest RMSSD. Led horses' responses to the cow and box were rated as more fearful as the distance to the stimulus decreased. Mares had a higher HR than geldings across all tests. HR positively correlated with the fearfulness rating at the furthest distance from the cow and box, and RMSSD negatively correlated with this rating in cow and control conditions. Our results show that these horses' avoidance response to cows was similar or higher to that shown towards a novel moving object, demonstrating that potentially, both neophobia and heterospecific communication play a role in this reaction.

Behavioural and Physiological Changes in a Herd of Arabian Mares after the Separation of Individuals Differently Ranked within the Dominance Hierarchy.

Horses in a herd develop and maintain a dominance hierarchy between all individuals. There are many situations in riding facilities and studs in which horses have to be separated out of a group. The aim of the study was to determine the rate of behaviours, level of locomotor activity and cardiac activity variables in a herd of horses during a short social separation of individuals differently ranked in the dominance hierarchy. Twelve adult Arabian mares were involved. A behavioural test had been performed before the main experiment to determine the rank order of the mares in this social herd. Three tests were performed when a dominant, mixed and submissive three-member group of mares was separated for 10 min. The response of the remaining herd was determined by a rate of behaviours, time of locomotor activity and cardiac parameters. The results of the experiment reveal evident changes towards emotional arousal in the social herd elicited by a short separation of some conspecifics. The herd created by humans preserves the sensitivity to a temporary loss of its members. The response of the remaining herd does not depend strictly on the composition of the separated mares regarding their rank in the dominance hierarchy.

Thermographic Analysis of the Metacarpal and Metatarsal Areas in Jumping Sport Horses and Leisure Horses in Response to Warm-Up Duration.

This study aimed to assess the impact of various types of warm-up on the metacarpal and metatarsal surface temperature in jumping sport horses in comparison to leisure horses, which work usually less intensively. Six clinically healthy sport geldings, contestants in showjumping competitions, and six geldings used for leisure riding were included in the study. The experiment was conducted for four consecutive days, during which the horses were warmed up by walking and trotting for various durations. Images were taken with a FLUKE Ti9 thermal imager to determine the resting, post-effort, and recovery temperature of the dorsal and plantar surface of the metacarpus and metatarsus of the four limbs. The obtained data were analysed with SmartView 4.1. software. The increase of measured rectal and surface temperatures was proportional to the warm-up duration. The surface temperature increase in the distal limb parts in jumping sport horses was greater than in horses used for leisure. The plantar surface was also warmer than the dorsal surface of the metacarpal/metatarsal areas, with a forelimb being warmer than a hind limb. Elevated temperatures after warm-up persist for 30 min in the recovery period, especially in jumping sport horses compared to leisure horses. Thus, the warming up effect is achieved earlier and lasts longer in heavily trained horses than in non-performance horses.

Plasma Activin A and Decorin in Exercised Purebred Arabian Horses - Preliminary Study.

Development of the musculoskeletal system occurs, in part, through an impact on regulatory proteins, such as activin A and decorin. Activin A induces atrophic effects on skeletal muscle, and decorin regulates the formation of connective tissue proteins like collagen and elastin in tendons and ligaments. The aim of this study was to evaluate the influence of different intensities of exercise on blood plasma activin A and decorin concentrations in horses. Ten young purebred Arabian horses were subjected to routine training sessions on a racetrack, and another 11 adult horses of the same breed participated in endurance rides. Race horses were examined during gallop tests over a distance of 1200 m and endurance horses at distances from 20 to 80 km. Blood samples were collected at rest and after exercise to determine plasma concentrations of activin A, decorin and cortisol. Despite differences in the intensity of exercise, the plasma decorin concentration remained unchanged, and activin A tended to decrease in endurance horses only. The exercise-induced changes in plasma activin A concentrations correlated with the covered distance (r = -0.43; P < .05), but not with the changes in cortisol values. Further studies are needed to confirm the usefulness of plasma activin A as a potential indicator of a horse's endurance performance.

Exploratory Behaviours of Primitive Horses Based on Konik: A Preliminary Study.

This study aimed at assessing the behaviour of Konik geldings and mares, kept in a stable and in a free-range system, during behavioural tests regarded as a determinant of the exploration urge. A total of 19 Konik horses kept in individual stables and in a free-range system were included in the study. The experiment was conducted in five phases separated by five-day breaks. A one-stage passive human test was performed during the first phase, a three-stage active human test-2nd phase, a three-stage unknown object test-3rd phase, a two-stage unknown surface test-4th phase, and a one-stage test of social isolation-5th phase. Ten attributes were analysed, including the horse sex and the keeping system. The results were also correlated with one another. Konik horses were found to show the urge to explore, although their behavioural responses are individual and stimulus dependent. In many cases, the horse sex and the keeping system influence the exploratory behaviour, although it is manifested by a greater intensity in geldings than in mares, and in free-range horses than in those kept in a stable. The study is regarded as preliminary due to the small number of horses in the study groups.

Horse Preferences for Insolation, Shade or Mist Curtain in the Paddock under Heat Conditions: Cardiac and Behavioural Response Analysis.

The horse's welfare and, consequently, the emotional arousal may be connected with stressful environmental conditions. This study aimed to determine whether horses show behavioural or physiological symptoms of thermal discomfort and if their behaviour and cardiac parameters are related to freely chosen insolated (IS), shaded (SH), or water sprayed (with a mist curtain (MC)) areas in a paddock under heat conditions (29-32 °C, 42.0 ± 1.5% humidity). Twelve adult horses freely moving in the paddock were studied during a 45 min solitary turnout. Six cardiac variables, locomotor, and non-locomotor activities as well as rectal temperature before and after the test were monitored with regard to the area of staying. Horses did not show clear preferences regarding the time spent in IS, SH, and MC, although preferences of particular horses differed considerably. When staying under IS and MC conditions, the horses showed a higher level of relaxation compared to SH. Horses did not exhibit symptoms of thermal discomfort while staying in the sun. Free choice between the three areas differing in environmental conditions could be a crucial factor in maintaining body temperature as well as emotional arousal at similar levels. Thus, the provision of a shade and mist curtain in paddocks seems to be reasonable.

Social Behaviour of Horses in Response to Vocalisations of Predators.

We tested the hypothesis that social defensive responses to the vocalisation of a predator still exist in horses. The recordings of a grey wolf, an Arabian leopard and a golden jackal were played to 20 Konik polski and Arabian mares. Durations of grazing, standing still, standing alert and the number of steps in walk and trot/canter were measured. In one-minute scans, the distances of the focal horse from the reference horse (DIST-RH) and from the nearest loudspeaker (DIST-LS) were approximated. The vocalisation of a leopard aroused the Arabians more than the Koniks (less grazing, stand-still and walk, more stand-alert and trotting/cantering). Koniks showed more relaxed behaviours to the leopard vocalisation (more grazing, stand-still and walk), but high alertness to the wolf playback (stand-alert, trotting/cantering). Spatial formation of the herd of Koniks showed tight grouping (lower DIST-RH) and maintaining distance from the potential threat (DIST-LS) in response to the wolf howling, while the Arabians approached the loudspeakers in linear herd formation when the leopard growls were played. Adult horses responded to potential predation by changing spatial group formations. This ability to apply a social strategy may be one of the explanations for the least number of horses among all hunted farm animal species.

Concentrations of Circulating Irisin and Myostatin in Race and Endurace Purebred Arabian Horses-Preliminary Study.

Skeletal muscle is considered to be the largest endocrine organ determining the maintenance of energy homeostasis. Adaptive changes in skeletal muscles in response to physical exercise influence the production as well as secretion of myokines, which are bioactive factors that play a crucial role in energy expenditure processes. The aim of the study was to investigate the impact of two different types of exercise on the circulating level of two of these, myostatin and irisin, in trained horses. Twenty purebred Arabian horses were involved in the study: 10 three-year-old horses trained on the racetrack and 10 endurance horses aged 7.4 ± 1.9 years. The horses from both groups were regularly trained throughout the entire season, during which they also participated in Polish National competitions. To assess the influence of the training sessions on plasma myostatin and irisin concentrations, blood samples taken at rest and 30 min after the end of exercise were analyzed. In the studied horses, the single bout of exercise did not influence plasma irisin but induced an increase in plasma myostatin concentration. In racehorses, plasma irisin concentration decreased with the length of the training season. Plasma myostatin was higher in endurance horses than in three-year-old racehorses. Lack of exercise-induced fluctuation in circulating irisin in studied horses suggests that myostatin released in response to exercise provides a negative feedback signal to irisin release.

The intensity of physiological and behavioral responses of horses to predator vocalizations.

BACKGROUND: Predatory attacks on horses can become a problem in some parts of the world, particularly when considering the recovering gray wolf populations. The issue studied was whether horses transformed by humans and placed in stable-pasture environments had retained their natural abilities to respond to predation risk. The objective of the study was to determine the changes in cardiac activity, cortisol concentrations, and behavior of horses in response to the vocalizations of two predators: the gray wolf (Canis lupus), which the horses of the breed studied had coevolved with but not been exposed to recently, and Arabian leopard (Panthera pardus nimr), from which the horses had been mostly isolated. In addition, we hypothesized that a higher proportion of Thoroughbred (TB) horse ancestry in the pedigree would result in higher emotional excitability in response to predator vocalizations. Nineteen horses were divided into groups of 75%, 50% and 25% TB ancestry. The auditory test conducted in a paddock comprised a 10-min prestimulus period, a 5-min stimulus period when one of the predators was heard, and a 10-min poststimulus period without any experimental stimuli. RESULTS: The increase in heart rate and saliva cortisol concentration in response to predator vocalizations indicated some level of stress in the horses. The lowered beat-to-beat intervals revealed a decrease in parasympathetic nervous system activity. The behavioral responses were less distinct than the physiological changes. The responses were more pronounced with leopard vocalizations than wolf vocalizations. CONCLUSIONS: The horses responded with weak signs of anxiety when exposed to predator vocalizations. A tendency towards a stronger internal reaction to predators in horses with a higher proportion of TB genes suggested that the response intensity was partly innate. The more pronounced response to leopard than wolf may indicate that horses are more frightened of a threatening sound from an unknown predator than one known by their ancestors. The differing response can be also due to differences in the characteristic of the predators' vocalizations. Our findings suggested that the present-day horses' abilities to coexist with predators are weak. Hence, humans should protect horses against predation, especially when introducing them into seminatural locations.

Bee Pollen Supplementation to Aged Horses Influences Several Blood Parameters.

In humans and animals, aging leads to a decrease in immune function and an increased susceptibility to infection. Decreased immunity and an increase in the incidence of infectious diseases are particularly notable during the autumn. Bee pollen supplementation improves immunity and antioxidant enzyme activity, as well as general performance. The aim of this study was to evaluate the effects of bee pollen supplementation during the autumn on blood parameters in aged horses. The study was performed on 16 warmblood horses aged 15-26 years. Half of this group received 60 g of bee pollen (soaked in water) daily for 30 days during the autumn season. Blood samples were taken from all horses before and after the supplementation period. Numerous hematological and plasma biochemical parameters including indicators of oxidative stress were determined. The data collected after the supplementation were compared with data collected before the experiment using one-way analysis of variance and paired Student's t-test. In the control group, there was a decline in the total number of red blood cells, hemoglobin, and hematocrit and an increase in some lipid parameters, urea, total plasma proteins, and sulfhydryl groups. Supplementation with bee pollen prevented the variation of these parameters, except for low-density lipoprotein cholesterol. We believe that bee pollen supplementation for aged horses during autumn has beneficial effects because it inhibited some of the adverse changes observed in the control horses during this season.

Distribution of Superficial Body Temperature in Horses Ridden by Two Riders with Varied Body Weights.

It was assumed that a horse with its rider body weight found in the upper limit may negatively impact the horse's welfare. The objective of this paper was to analyze the differences in body temperature and selected heart rate parameters in horses in response to physical exercise accompanied by various rider's body weight loads. The study was carried out on 12 leisure, 10-15-year-old warmblood geldings. The horses were ridden by two equally qualified riders whose body weights were about 20% and 10% of the average body weight (BW) of the animals (about 470 kg). Each rider rode each of the 12 horses for 13 min walking and 20 min of trotting. Images of the horse at rest, after physical exercise directly after unsaddling, and during the recovery phase (10 min after unsaddling) were taken with an infrared thermography camera. For analysis, the temperatures of selected body parts were measured on the surface of the head, neck, front, middle, and back (croup) parts of the trunk, forelimb, and hind limb. Immediately after the infrared thermography images were taken, the rectal temperature of the horse was measured. The heart rate parameters were measured at rest for 10 min directly before, during, and 10 min following the end of a training session. A multivariate analysis of variance (ANOVA) for repeated measurements was performed. Statistical significance was accepted for p < 0.05. A rider BW load on a horse of approximately 20% of the horse's BW led to a substantial increase in the superficial temperatures of the neck, front, middle, and back parts of the trunk in relation to these body parts' average temperatures when the load was about 10% BW. The head and limb average temperatures were not significantly affected by the load of the exercised horse. A horse's load above 20% of his body weight, even with little effort, affects changes in surface temperature and the activity of the autonomic nervous system.