An exploratory study of competition scores and salivary cortisol concentrations in Warmblood horses.

The main objective of this explorative study was to describe the relationship between competition scores and salivary cortisol concentrations in young horses during dressage and showjumping competitions. The study also investigated whether the diurnal rhythm of salivary cortisol concentrations was affected by competition over consecutive days compared with the home environment. Saliva samples were collected from 126 dressage horses and showjumpers in their home environment and at 3 different events. The relationship between scores given by judges at the competition and cortisol concentrations at the event was assessed. The results demonstrated that competition scores correlated positively to baseline cortisol concentrations at one of 3 events (r = 0.53, P < 0.001). Salivary cortisol concentrations followed a diurnal rhythm with the highest concentrations measured in the morning and the lowest in the evening, both at home and in the competition environment (P < 0.05). Salivary cortisol concentrations were greater during the competitions than at home (P < 0.05) except at one event where showjumpers did not increase between home and competition. Dressage horses had the highest baseline cortisol concentrations at competition, and exercise caused cortisol concentrations to increase in both showjumpers and dressage horses (P < 0.001). In conclusion, the diurnal rhythm in salivary cortisol concentrations was maintained in the novel environment. Dressage horses demonstrated greater baseline cortisol concentrations at competition than showjumpers, suggesting that they may perceive the novel environment as more stressful. Furthermore, there was no consistent relationship between baseline salivary cortisol concentrations and competition scores across the events.

A Nordic survey of management practices and owners' attitudes towards keeping horses in groups.

Keeping horses in groups is widely recommended but limited information is available about how this is implemented in practice. The aim of this survey was to describe how horses are kept in the Nordic countries in relation to sex, age, breed, and equestrian discipline and to assess owners' attitudes toward keeping horses in groups. Horse owners in Denmark, Finland, Norway, and Sweden were approached using a web-based questionnaire, which was translated into 4 languages and distributed online via equestrian forums, organizations, and social media. The number of respondents was 3,229, taking care of 17,248 horses. Only 8% of horses were never kept in groups, 47% were permanently grouped for 24 h/d, and 45% were stabled singly but grouped during turnout. Yearlings were most often permanently kept in groups (75%), mares and geldings more commonly during parts of the day (50 and 51%, respectively), and stallions were often kept alone (38%). Icelandic horses were more likely to be permanently kept in groups (36%) than warmbloods (16%) and ponies (15%). Twice as many competition horses (51%) were never grouped compared with horses used for breeding (20%) or leisure purposes (15%). The majority of respondents (86%) strongly agreed that group housing benefits horse welfare and that it is important for horses to have the company of conspecifics (92%). Nevertheless, not all horses were kept in groups, showing that attitudes toward group housing may not necessarily reflect current management. The risk of injury was a concern of many respondents (45%), as was introducing unfamiliar horses into already established groups (40%) and challenges in relation to feeding in groups (44%). Safety of people (23%) and difficulties handling group-kept horses (19%) were regarded as less problematic. Results suggest that the majority of horses have the possibility to freely interact with other horses, either as fulltime members of a group during 24 h/d or during turnout. Future research should address the extent to which being a part-time member of a group affects horse welfare. For permanent group housing to become more widespread, such as it is the case for most farm animals, future research could focus on solving some of the reoccurring problems perceived with keeping horses in groups. The dissemination of evidence-based information on all aspects around keeping horses in groups can ultimately stimulate further positive changes in the management of group-kept horses.

Two- and three-dimensional transvaginal ultrasound with power Doppler angiography and gel infusion sonography for diagnosis of endometrial malignancy.

OBJECTIVES: To evaluate the diagnostic efficiency of two-dimensional (2D) and three-dimensional (3D) transvaginal ultrasonography, power Doppler angiography (PDA) and gel infusion sonography (GIS) at offline analysis for recognition of malignant endometrium compared with real-time evaluation during scanning, and to determine optimal image parameters at 3D analysis. METHODS: One hundred and sixty-nine consecutive women with postmenopausal bleeding and endometrial thickness ≥ 5 mm underwent systematic evaluation of endometrial pattern on 2D imaging, and 2D videoclips and 3D volumes were later analyzed offline. Histopathological findings at hysteroscopy or hysterectomy were used as the reference standard. The efficiency of the different techniques for diagnosis of malignancy was calculated and compared. 3D image parameters, endometrial volume and 3D vascular indices were assessed. Optimal 3D image parameters were transformed by logistic regression into a risk of endometrial cancer (REC) score, including scores for body mass index, endometrial thickness and endometrial morphology at gray-scale and PDA and GIS. RESULTS: Offline 2D and 3D analysis were equivalent, but had lower diagnostic performance compared with real-time evaluation during scanning. Their diagnostic performance was not markedly improved by the addition of PDA or GIS, but their efficiency was comparable with that of real-time 2D-GIS in offline examinations of good image quality. On logistic regression, the 3D parameters from the REC-score system had the highest diagnostic efficiency. The area under the curve of the REC-score system at 3D-GIS (0.89) was not improved by inclusion of vascular indices or endometrial volume calculations. CONCLUSION: Real-time evaluation during scanning is most efficient, but offline 2D and 3D analysis is useful for prediction of endometrial cancer when good image quality can be obtained. The diagnostic efficiency at 3D analysis may be improved by use of REC-scoring systems, without the need for calculation of vascular indices or endometrial volume. The optimal imaging modality appears to be real-time 2D-GIS.

Effects of handling on fear reactions in young Icelandic horses.

REASONS FOR PERFORMING THE STUDY: Inclusion of objective temperament tests at practical horse breeding evaluations is of increased interest. It has been debated whether such tests may involve human handling, since there may be considerable differences in horses' handling experience. OBJECTIVES: To investigate the effect of a short-term standardised handling procedure on reactions of young horses in 2 types of fear tests (including and excluding human handling). STUDY DESIGN: An experimental study with 3-year-old Icelandic horses (n = 24). METHODS: Handled horses (n = 12) were trained according to a standardised handling procedure whereas controls (n = 12) remained untrained. Behavioural and heart rate responses in a novel object test and 2 handling fear tests (HFTs) were measured. The HFTs were conducted with both an unknown (HFT-unknown) and a known handler (HFT-known). RESULTS: There was no effect of the handling procedure on the horses' behavioural and heart rate responses in the novel object test, nor in the HFT-unknown. In the HFT-known, however, handled horses showed a significantly shorter duration of reluctance behaviour compared with controls, whereas heart rates did not differ. Heart rates correlated significantly between tests. CONCLUSIONS: Previous handling may affect the behavioural fear response of horses when handled by their usual handler, whereas this effect did not apply to an unknown handler. Heart rates appeared unaffected by handling and may be a more reliable indicator of fearfulness. Known handlers may 'mask' behavioural responses of horses in fear tests and thus handling by a known handler during testing may not be appropriate for objective evaluation of fearfulness in a practical situation.

Effects of hyperflexion on acute stress responses in ridden dressage horses.

The effects of hyperflexion on the welfare of dressage horses have been debated. This study aimed to investigate acute stress responses of dressage horses ridden in three different Head-and-Neck-positions (HNPs). Fifteen dressage horses were ridden by their usual rider in a standardised 10-min dressage programme in either the competition frame (CF), hyperflexion ("Low-Deep-and-Round"; LDR) or a looser frame (LF) in a balanced order on three separate test days. Heart rate (HR), heart rate variability parameters (HRV), behaviour and rein tension were recorded during the test. Salivary cortisol concentrations were measured 60 min before and 0, 5, 15 and 30 min after the test. Rein tension was significantly lower in LF and did not differ between CF and LDR; however approx. 15% of recordings in CF and LDR were above the sensor detection limit of 5 kg. The horses had significantly higher cortisol concentrations directly after LDR compared to LF. In addition, the horses showed more distinctive head movements, including head waving, during LDR. There were no significant treatment effects on HR and HRV. In conclusion, the results indicate that LDR may be more stressful to these horses during riding.

Object habituation in horses: the effect of voluntary versus negatively reinforced approach to frightening stimuli.

REASONS FOR PERFORMING THE STUDY: The ability of horses to habituate to novel objects influences safety in the horse-human relationship. However, the effectiveness of different habituation techniques has not been investigated in detail. OBJECTIVES: 1) To investigate whether horses show increased stress responses when negatively reinforced to approach novel objects, compared with horses allowed to voluntarily explore the objects and 2) whether a negatively reinforced approach facilitates object habituation. METHODS: Twenty-two 2-3-year-old Danish Warmblood geldings were included. Half of the horses (NR group) were negatively reinforced by a familiar human handler to approach a collection of novel objects in a test arena. The other half were individually released in the arena and were free to explore the objects (VOL group). On the next day, the horses were exposed to the objects again without a human handler, to investigate the rate of habituation. Behavioural and heart rate responses were recorded. RESULTS: All VOL horses initially avoided the unknown objects, whereas the handler was able to get all NR horses to approach and stand next to the objects within the first 2 min session. The NR horses had a significantly longer duration of alertness and a higher max heart rate in the first session. On the next day, however, NR horses spent significantly less time investigating the objects and had a shorter latency to approach a feed container, placed next to the objects, indicating increased habituation. CONCLUSION: A negatively reinforced approach to novel objects increases stress responses during the initial exposure but facilitates habituation in young horses. POTENTIAL RELEVANCE: Although a negatively reinforced approach appears beneficial for habituation, the procedure should be carefully managed due to increased stress responses in the horse, which may constitute a safety risk. Further experiments should aim to investigate differences in stimulus intensity.

Rein tension acceptance in young horses in a voluntary test situation.

REASONS FOR PERFORMING THE STUDY: During riding, horses are frequently exposed to pressure from the rider, e.g. through the bit and reins, but few studies have investigated at which point rein tension becomes uncomfortable for the horse. OBJECTIVES: To investigate how much rein tension young inexperienced horses are willing to accept in order to obtain a food reward; whether the tension acceptance changes during 3 consecutive test days; and whether rein tension correlates with the expression of conflict behaviour and heart rate. HYPOTHESES: Pressure-naïve horses will apply only little rein tension in the first voluntary trial, but their acceptance will gradually increase. High levels of rein tension will lead to expression of conflict behaviour and increases in heart rate. METHODS: Fifteen 2-year-old, bridle-naïve mares were encouraged to stretch their head forward (across a 0.95 m high metal bar) to obtain a food reward in a voluntary test situation. On each test day, each horse was exposed to 2 control sessions (loose reins), an intermediate and a short rein session (1 min/session). Rein tension, heart rate and behaviour were recorded. RESULTS: The horses applied significantly more tension on the first day (mean rein tension: 10.2 N), compared to the second and third test day (Day 2: 6.0 and Day 3: 5.7 N). The horses showed significantly more conflict behaviour in the short rein treatment. There was no treatment effect on heart rate. CONCLUSIONS AND POTENTIAL RELEVANCE: The horses applied the highest rein tension on the first day, and apparently learned to avoid the tension, rather than habituate to it. Rein tension correlated with expression of conflict behaviour, indicating that the horses found the tension aversive. Further studies should focus on the correlation between rein tension and conflict behaviour in ridden horses.

Training young horses to social separation: effect of a companion horse on training efficiency.

REASONS FOR PERFORMING STUDY: The intensity with which a horse responds to separation from its group and subsequently to being alone is relevant for both horse and handler safety. Identification of training methods that may reduce responses to separation would be useful in practice. OBJECTIVES: To investigate whether the initial presence of a familiar companion horse modifies responses to separation from the group, lowers stress levels (as measured by heart rate) and increases training efficiency. HYPOTHESIS: Habituation to separation proceeds more quickly if the horse is first trained with a companion, and heart rate is lower when the horse is subsequently trained alone, compared to control horses trained individually from the start. METHODS: Young mares (n = 32), kept in groups of 4 were exposed to social separation: 2 horses of the group were trained singly (S1, n = 16) and the remaining 2 horses (n = 16) were trained first with a companion (P2) and then alone (P1). The training comprised 3 steps whereby distance from the group was gradually increased. The final learning criterion was met when a horse fed calmly alone inside a test arena (Step 3). Horses that were trained in a pair had to succeed in Step 3 together before they repeated the steps alone. Feeding behaviour and heart rate were recorded. RESULTS: There were no significant differences between S1 and P1 horses, indicating that the initial pair-training did not reduce responses in P1 horses. However, heart rate was significantly lower when horses were trained in pairs (P2) compared to when the same horses were subsequently trained alone (P1). CONCLUSIONS AND POTENTIAL RELEVANCE: It may not be efficient to habituate naïve young horses to social separation initially with a partner as these horses appear to have to relearn being in the test situation alone when switching to the individual training.

Effects of a calm companion on fear reactions in naive test horses.

REASON FOR PERFORMING STUDY: In fear-eliciting situations, horses tend to show flight reactions that can be dangerous for both horse and man. Finding appropriate methods for reducing fearfulness in horses has important practical implications. OBJECTIVES: To investigate whether the presence of a calm companion horse influences fear reactions in naive subject horses. HYPOTHESES: The presence of a habituated (calm) companion horse in a fear-eliciting situation can reduce fear reactions in naive subject horses, compared to subject horses with a nonhabituated companion (control). METHODS: Minimally handled (n = 36), 2-year-old stallions were used, 18 as subjects and 18 as companions. Companion horses (n = 9) were habituated to an otherwise frightening, standardised test stimulus (calm companions), whereas the rest (n = 9) of the companion horses remained nonhabituated (control companions). During the test, unique pairs of companion and subject horses were exposed to the test stimulus while heart rate and behavioural responses were registered. Subsequently, subject horses were exposed to the stimulus on their own (post test). RESULTS: Subject horses, paired with a calm companion horse, showed less fear-related behaviour and lower heart rate responses compared to subject horses with control companions. Results from the post test suggest that the difference between treatment groups remained in the subsequent absence of companion horses. CONCLUSIONS AND POTENTIAL RELEVANCE: It appears possible to reduce fear reactions in young, naive horses by allowing them to interact with a calm companion horse in fear-eliciting situations.

Training methods for horses: habituation to a frightening stimulus.

REASONS FOR PERFORMING STUDY: Responses of horses in frightening situations are important for both equine and human safety. Considerable scientific interest has been shown in development of reactivity tests, but little effort has been dedicated to the development of appropriate training methods for reducing fearfulness. OBJECTIVES: To investigate which of 3 different training methods (habituation, desensitisation and counter-conditioning) was most effective in teaching horses to react calmly in a potentially frightening situation. HYPOTHESES: 1) Horses are able to generalise about the test stimulus such that, once familiar with the test stimulus in one situation, it appears less frightening and elicits a reduced response even when the stimulus intensity is increased or the stimulus is presented differently; and 2) alternative methods such as desensitisation and counter-conditioning would be more efficient than a classic habituation approach. METHODS: Twenty-seven naive 2-year-old Danish Warmblood stallions were trained according to 3 different methods, based on classical learning theory: 1) horses (n = 9) were exposed to the full stimulus (a moving, white nylon bag, 1.2 x 0.75 m) in 5 daily training sessions until they met a predefined habituation criterion (habituation); 2) horses (n = 9) were introduced gradually to the stimulus and habituated to each step before the full stimulus was applied (desensitisation); 3) horses (n = 9) were trained to associate the stimulus with a positive reward before being exposed to the full stimulus (counter-conditioning). Each horse received 5 training sessions of 3 min per day. Heart rate and behavioural responses were recorded. RESULTS: Horses trained with the desensitisation method showed fewer flight responses in total and needed fewer training sessions to learn to react calmly to test stimuli. Variations in heart rate persisted even when behavioural responses had ceased. In addition, all horses on the desensitisation method eventually habituated to the test stimulus whereas some horses on the other methods did not. CONCLUSIONS AND POTENTIAL RELEVANCE: Desensitisation appeared to be the most effective training method for horses in frightening situations. Further research is needed in order to investigate the role of positive reinforcement, such as offering food, in the training of horses.

Behavioural responses of farm mink towards familiar and novel food.

The study aimed to (i) describe the response of farmed mink towards familiar and novel food, and (ii) assess the suitability of using novel food in order to measure fear responses in mink. A total of 48 farm mink from two behaviourally selected genetic lines were caged individually with one standardised daily feeding. The experimental feedings were carried out in a balanced order on 2 successive days. Behaviour of the mink was video recorded for 10min after food provision. In general, the mink were reluctant to approach and eat the novel food compared to familiar food, which indicates a neophobic feeding strategy, i.e. increased latency to come close to (P<0.01) and eat (P<0.001) the novel food, and a higher frequency (P<0.001) and a longer duration (P<0.001) of sniffing. Provision of unknown food items may be used to evaluate fear responses in mink since mink respond reluctantly towards novel food. However, high inter-individual variation in the present study obliterated a previously shown difference between confident and fearful mink, suggesting that this method for measuring fear responses in mink is appropriate only on a large animal material.