Manuela Wulf

Physiological and behavioural responses of young horses to hot iron branding and microchip implantation.

Branding is the traditional and well-established method used to mark horses, but recently microchip transponders for implantation have become available. In this study, behaviour, physiological stress variables and skin temperature in foals were determined in response to hot-iron branding (n=7) and microchip implantation (n=7). Salivary cortisol concentrations increased in response to branding (1.8 ± 0.2 ng/mL) and microchip implantation (1.4 ± 0.1ng/mL), but cortisol release over time did not differ. In response to both manipulations there was a transient increase in heart rate (P<0.001) and heart rate variability (P<0.01). Branding and microchip implantation induced a comparable aversive behaviour (branding, score 3.86 ± 0.85; microchip, score 4.00 ± 0.82). Both techniques thus caused similar physiological and behavioural changes indicative of stress. Acutely, implantation of a microchip was as stressful as branding in foals. Branding caused a necrotising skin burn lasting at least 7 days. Moreover branding, but not microchip implantation (P<0.001), was accompanied by a generalized increase in skin temperature which was comparable to low degree post-burn hypermetabolism in humans.

Cortisol and progestin release, heart rate and heart rate variability in the pregnant and postpartum mare, fetus and newborn foal

The mechanisms leading to parturition in the horse in many aspects differ from those in other species. Pregnancy is maintained not by progesterone but by 5α-pregnanes and the progestin precursor pregnenolone originates from the fetus. As parturition approaches, the fetal adrenal switches from pregnenolone to cortisol synthesis but it is not known whether cortisol crosses the placenta. We hypothesized that in parallel to fetal cortisol release, cortisol in the maternal circulation increases before foaling and this increase can be determined in both saliva and plasma. In addition, maternal, fetal and neonatal heart rate and heart rate variability were measured. In 25 pregnant mares, saliva for cortisol analysis was collected 4 times daily from 15 days before to 5 days after foaling. In 13 mares, in addition, fetomaternal electrocardiogram (ECG) recordings were made and blood samples for progestin and cortisol analysis were collected once daily. Heart rate (HR) was recorded until 5 days after foaling. The heart rate variability (HRV) variables standard deviation of the beat-to-beat (RR) interval (SDRR) and root mean square of successive RR differences (RMSSD) were calculated. From Days 15 to 4 before parturition, progestin concentration increased (peak 267 ± 42 ng/mL) and decreased thereafter (P < 0.05, day of foaling 113 ± 18 ng/mL). A prepartum increase in maternal cortisol concentrations was evident in blood (P < 0.05) and saliva (P < 0.05) and paralleled the decrease in progestin concentrations. In mares, HR remained constant during the last days of pregnancy but decreased within one day after parturition (P < 0.05) while maternal HRV did not change. In the fetus and neonate, HR increased from before to after birth (P < 0.05) indicating increasing demands on the cardiovascular system with adaptation to extrauterine life.

Behavioral and physiological responses of young horses to different weaning protocols: a pilot study.

In this study, effects of weaning on behavioral and physiological stress parameters in young horses (foals) were determined. Foals were weaned either simultaneously without the presence of adult horses (group A, n = 6), or in the presence of two adult females familiar but unrelated to the foals (group B, n = 5), or weaned consecutively by removing two mother horses per day (group C, n = 6). Behavior, locomotion, salivary cortisol concentration, beat-to-beat (RR) interval, heart rate variability (HRV) and weight were determined. Group A foals lost weight for 2 days (mean ± SEM) − 8.3 ± 1.6 kg, p < 0.05. Weaning was followed by increased vocalization which was least pronounced in foals of group B (p < 0.05). Locomotion was most pronounced on weaning day in foals of group A and lowest in group B (p < 0.05). Weaning increased salivary cortisol concentration on the day of weaning in groups A and B and for 2 days in group C (p < 0.05). The RR interval decreased most pronouncedly in group A foals (p < 0.05). There were no consistent changes in HRV. Based on cortisol release and behavior, weaning is associated with stress but this was least pronounced in foals weaned in the presence of two familiar but unrelated adult female horses.

Cortisol release, heart rate and heart rate variability, and superficial body temperature, in horses lunged either with hyperflexion of the neck or with an extended head and neck position

Bringing the head and neck of ridden horses into a position of hyperflexion is widely used in equestrian sports. In our study, the hypothesis was tested that hyperflexion is an acute stressor for horses. Salivary cortisol concentrations, heart rate, heart rate variability (HRV) and superficial body temperature were determined in horses (n = 16) lunged on two subsequent days. The head and neck of the horse was fixed with side reins in a position allowing forward extension on day A and fixed in hyperflexion on day B. The order of treatments alternated between horses. In response to lunging, cortisol concentration increased (day A from 0.73 ± 0.06 to 1.41 ± 0.13 ng/ml, p < 0.001; day B from 0.68 ± 0.07 to 1.38 ± 0.13 ng/ml, p < 0.001) but did not differ between days A and B. Beat-to-beat (RR) interval decreased in response to lunging on both days. HRV variables standard deviation of RR interval (SDRR) and RMSSD (root mean square of successive RR differences) decreased (p < 0.001) but did not differ between days. In the cranial region of the neck, the difference between maximum and minimum temperature was increased in hyperflexion (p < 0.01). In conclusion, physiological parameters do not indicate an acute stress response to hyperflexion of the head alone in horses lunged at moderate speed and not touched with the whip. However, if hyperflexion is combined with active intervention of a rider, a stressful experience for the horse cannot be excluded.

Effects of season, age, sex, and housing on salivary cortisol concentrations in horses

Analysis of salivary cortisol is increasingly used to assess stress responses in horses. Because spontaneous or experimentally induced increases in cortisol concentrations are often relatively small for stress studies, proper controls are needed. This requires an understanding of the factors affecting salivary cortisol over longer times. In this study, we have analyzed salivary cortisol concentration for 6 mo in horses (n = 94) differing in age, sex, reproductive state, and housing. Salivary cortisol followed a diurnal rhythm with the highest concentrations in the morning and a decrease throughout the day (P < 0.001). This rhythm was disrupted in individual groups on individual days; however, alterations remained within the range of diurnal changes. Comparison between months showed highest cortisol concentrations in December (P < 0.001). Cortisol concentrations increased in breeding stallions during the breeding season (P < 0.001). No differences in salivary cortisol concentrations between nonpregnant mares with and without a corpus luteum existed. In stallions, mean daily salivary cortisol and plasma testosterone concentrations were weakly correlated (r = 0.251, P < 0.01). No differences in salivary cortisol between female and male young horses and no consistent differences between horses of different age existed. Group housing and individual stabling did not affect salivary cortisol. In conclusion, salivary cortisol concentrations in horses follow a diurnal rhythm and are increased in active breeding sires. Time of the day and reproductive state of the horses are thus important for experiments that include analysis of cortisol in saliva.

Effects of the level of experience of horses and their riders on cortisol release, heart rate and heart-rate variability during a jumping course

Equestrian sports require the co-operation of two species, horses and humans, but it is unknown to what extent stress responses in the rider affect the horse. In this study, the stress response of experienced and less-experienced horses and riders at showjumping was analysed. Sixteen sport horses were divided into two groups (n = 8 each) by experience and were ridden by highly experienced professionals (n = 8) and less-experienced riders (n = 8). Riders jumped a course of obstacles with an experienced and a less-experienced horse and horses took part with an experienced and less-experienced rider. Salivary cortisol, heart rate and heart-rate variability (HRV) variables, standard deviation of RR interval (SDRR) and root mean square of successive RR differences (RMSSD) were analysed. Cortisol and heart rate increased and HRV decreased in all riders and horses. In less-experienced riders, cortisol release was higher on a lessexperienced versus an experienced horse but the horses’ cortisol release was not affected by experience of their riders. Heart rate did not differ between groups of horses and was not affected by experience of the rider but was higher in less-experienced versus experienced riders. The HRV decreased in horses and riders and SDRR was lower in less-experienced versus experienced riders. Thus, lower experience of riders appears not to affect physiological stress parameters in their horses during a showjumping course.

Influence of semen collection on salivary cortisol release, heart rate, and heart rate variability in stallions

The aim of this study was to evaluate the stress response of stallions (n = 16) aged 3-13 years with previous sexual experience to semen collection by determination of heart rate, heart rate variability, and cortisol in saliva. Recordings were done on two consecutive days. The time intervals from leaving the box until arrival in the collection barn and from first exposure to the teaser mare until ejaculation as well as the number of mounts until ejaculation were neither affected by collection day nor by age, sexual experience (i.e., the number of breeding seasons the stallion experienced), or sexual workload of the stallion (i.e., the mean number of semen collections per week). Heart rate was continuously determined from 30 minutes before to 30 minutes after ejaculation and significantly increased in response to the semen collection procedure (P < 0.001). Changes in heart rate were significantly influenced by sexual experience (P < 0.01) and sexual workload (P < 0.05) but not by the age of the stallions. Day of semen collection did not have any effects. The heart rate variability variable root mean square of successive RR differences was not affected by semen collection procedures. Cortisol concentration in saliva was determined from 60 minutes before to 120 minutes after ejaculation and did not change significantly. The results indicate that semen collection is perceived as not more than a modest temporary stressor in sexually experienced and well-trained stallions.

Development of the genital microflora in stallions used for artificial insemination throughout the breeding season

An important factor influencing stallion fertility is the microbial contamination of semen. Aims of this study were to investigate changes in the microbiological population of the genital mucosa and semen in artificial insemination stallions (n=16) from before to after one breeding season (February-August). MALDI-TOF-MS (matrix assisted laser desorption/ionization time-of-flight mass spectrometry) was used for identification of microbial agents. For bacteriology, swabs from the urethral opening, urethral fossa and penile sheath as well as semen were collected at 4-week-intervals. For semen motility and percentage of morphologically normal spermatozoa, changes over time (P<0.001) occurred. In 14.3% of genital swabs and 25.0% of ejaculates no microbial growth was found. Intensity of total microbial growth increased throughout the breeding season (P<0.001). From the penile sheath, between 1.4±0.1 microbial species in February and 3.3±0.4 in August were identified. From semen, 1.1±0.3 microbial species in February and 2.9±0.6 in August were obtained. The number of microbial species isolated from the sheath of the penis (2.0±0.1) and urethral fossa (2.0±0.1) was greater (P<0.01) compared with the urethral opening (1.6±0.1) and semen (1.5±0.1). The microbial flora consisted of aerobic and anaerobic bacteria, dominated by coagulase-negative staphylococci, alpha-haemolytic streptococci and coryneforms. Only occasionally potentially pathogenic agents (E. coli, Ps. aeruginosa) were found. The microbial flora was not related to seminal characteristics.

Heart rate and salivary cortisol concentrations in foals at birth.

Heart rate (HR), HR variability (HRV) and salivary cortisol concentrations were determined in foals (n = 13) during the perinatal phase and until 5 months of age. In the fetus, HR decreased from 77 ± 3 beats/min at 120 min before birth to 60 ± 1 beats/min at 5 min before birth (P <0.01). Within 30 min of birth, HR increased to 160 ± 9 beats/min (P <0.01). Salivary cortisol concentrations immediately after birth were 11.9 ± 3.6 ng/mL and within 2 h increased to a maximum of 52.5 ± 12.3 ng/mL (P <0.01). In conclusion, increases in HR and salivary cortisol concentrations in foals are not induced during parturition, but occur immediately after birth.

Physiological stress responses and horse rider interactions in horses ridden by male and female riders

Traditionally, horse riding has been restricted to men but today equestrian sports are dominated by women. We hypothesised that men and women differ with regard to riding and the response they evoke in their horse. Cortisol and heart rate variability (HRV) were studied in male (n=8) and female riders (n=8) and in horses (n=8) ridden by men and women over a jumping course. Saliva for cortisol analysis was collected, cardiac beat to beat (RR) intervals were recorded and heart rate and HRV variables SDRR (standard deviation of RR interval) and RMSSD (root mean square of successive RR differences) calculated. In another experiment, saddle pressure was compared between male and female riders (n=5 each). Cortisol did not differ between male and female riders and increased in horses (P<0.001) irrespective of the sex of the rider. Heart rate in riders increased from walk to jumping (P<0.001) while HRV decreased (P<0.001) to the same extent in men and women. In horses, heart rate increased (P<0.001) and SDRR and R...