Abstract: Reasons for performing study: Little information exists on the immunological effects of transport or the use of supplements to minimise transport stress. Objectives: To establish baseline ranges and evaluate immunophenotypic and functional changes associated with transport and a nutritional ‘adaptogen’ supplement. Methods: Horses received either supplement (n = 10) or placebos (n = 9) during the 30 day study. After 28 days in stalls, 12 horses (6 supplement; 6 placebo) were transported for 24 h, then unloaded and recovered. Venous blood samples were collected on Days 1, 14 and 28 to establish baselines, and on Days 28, 29 and 30 to examine changes during transport and recovery. Results: Transport prompted elevations (P<0.05) in cortisol concentration, neutrophil count and white blood cell counts, while lymphocyte subpopulation counts (CD3+, CD4+, CD8+, CD21+) decreased (P<0.05). Normal phenotypic lymphocyte profiles returned within 24 h of recovery. Supplement effects on immunophenotype (CD21+ and CD8+) were observed in stabled horses (P<0.05), but not in transported horses. Conclusions: These results provide insights into the immunological mechanisms associated with long-term transport. Potential relevance: The existence of a small window of immunological uncertainty follows long-term transportation, enhancing the potential risk of infectious disease in susceptible individuals.

Abstract: In order to improve the housing conditions of stallions in individual boxes by offering a possibility to have more social contact, the Swiss national stud farm tested a new box system for horses, allowing increased physical contact with the neighbouring stallion. The aim of this part of the study was to investigate whether this type of housing system (named “social box”) potentially induces a change in stress reactions in stallions compared to conventional boxes. Therefore faecal cortisol metabolite (FCM) concentration was measured as a non-invasive parameter to assess endocrine responses related to this new environment.
Four groups each consisting of eight adult Freiberger breeding stallions were included in the test design. Every stallion spent three weeks in a conventional box and in a social box respectively (cross-over design). The conventional box consisted of a separation wall with a lower opaque part and an upper part with vertical barriers (5 cm between barriers), allowing visual and olfactory contact but strongly limiting tactile contact. The separating wall of the social box consisted of two lateral sections, one part being opaque to the ceiling and the second part consisting of vertical barriers (30 cm between barriers), allowing the horse to have physical contact with its neighbour or to avoid it.
In horses, FCM concentration reflects an average level of circulating cortisol over a period of approximatively 24h. Faecal samples were collected the day following integration in social / conventional boxes, reflecting the potential stress induced by increased social interactions during the integration. In order to asses potential chronical stress, faeces samples were also collected in week one, two and three after the integration into the social / conventional box (in total: 4 samples per horse and housing system). The samples were immediately stored at -20°C until they were analysed. The samples were not analysed in the laboratory until the end of the experiment, therefore the duration of conservation in the freezer varied from 40 to 429 days.
A considerable percentage of data from groups 1 and 2 was below the detection limit (<0.8 ng/g) (Tab. 1). Thus the statistical analysis was conducted with the FCM concentration from groups 3 and 4 (n horses = 16) which contained no values below the detection limit.
Tab. 1: Details about FCM values and storage time for the 4 groups of stallions
Group Storage duration [d] Proportion of data below the detection limit (<0.8 ng/g) Mean [ng/g] Median [ng/g]
Group 1 384-429 55.6 % 2.2 0
Group 2 315-360 25.5 % 5.8 6.3
Group 3 41-79 0.0 % 8.7 8.0
Group 4 40-85 0.0 % 5.8 5.4
Despite the impressive social interactions observed between the stallions directly after being introduced into the social boxes, we did not find any differences in FCM concentration between the stallions being introduced into the conventional box and the social box on the day of integration (social box: n samples = 16, mean±SD: 6.9±4.7 ng/g; conventional box: n samples = 16, mean±SD: 9.0±11.2 ng/g; Wilcoxon signed rank test V = 70, p = 0.94).
Overall the samples taken during integration and in week one, two and three did not show evidence of changes in FCM concentration in either housing system over a longer period of time (social box: n samples = 64, mean±SD: 7.9±6.2 ng/g; conventional box: n samples = 64, mean±SD: 6.6±3.4 ng/g; Linear mixed model (LMM), p = 0.56).
Our results suggest that the possibility of having physical contact with a conspecific does not induce changes in FCM concentration in breeding stallions. The considerable percentage of values below the detection limit in groups 1 and 2 seemed to correlate with the increasing duration of storage before analysis. During the IESM Network Meeting 2015, we would like to discuss possible methodological issues and the possibilities to correctly integrate these low values in the statistical analysis.

Abstract: According to Interpersonal Theory every interaction is motivated by efforts to achieve and maintain self-esteem and to avoid anxiety. People"s characteristic ways of accomplishing these ends are called interpersonal reflexes. Those interpersonal reflexes are evident in interaction with animals, since they are determined by the interpersonal traits of personality. We wanted to catch the typical interpersonal reflexes in between humans and horses compared to pet animals.
We used the self rating assessment instrument “Inventory of Problematic Interactions with Animals” (IPI – Animals), which bases on a Interpersonal Circumplex Model (Human Animal Circumplex; HAC) and was constructed to catch specific dispositions of distress caused by animals using two dimensions (too dominant vrs too submissive and too warm versus too cold). Data of 233 male adolescents (93 of them actual pet owners, from that 12 horse owners) were collected.
We found that different pet preferences holds distinct locations in the HAC. Horse persons differ from dog and cat persons within the dimension dominance (dog: chi2(df126) =161.54 p= .018; cat: chi2(df126) =199.95 p= .045). Persons, who own a horse or would wish to own one, describe themselves as dominant, but warm interactors. They report that they want horses to notice them. They tend do too much for them and behave very effusively with them. On the other hand they feel that the animal takes too much advantage of the relationship.

Abstract: Domestic horses are kept in so-called “fate societies” where they have to deal with frequent mixing. Several studies have evaluated and discussed the aggression level and injury risk during the introduction of horses into new groups, but nothing is known about the endocrine responses and thus if horses experience stress during introduction.
In this study we analysed the efficiency of four approved introduction techniques and evaluated the introduction of 30 horses into 11 different groups. Horses were introduced: 1) immediately, 2) after observing the new group for several days, 3) together with an “integration horse” after several days of observation, or 4) with a mixed strategy. Aggressive as well as positive social behaviour between the introduced horses and the group members were analysed the two hours following the introduction event. In addition, we focussed on the glucocorticoid production of the newcomer horses by measuring faecal cortisol metabolites (FCM) on the day of the introduction as well as the following three days.
For the four introduction techniques we found significant differences in the horses’ aggressive and submissive behaviour as well as in their total interactions. The introduction together with an integration horse led to significantly lower levels of aggression and less total interactions than the immediate introduction of single horses.
Horses which were introduced immediately or after an observation period showed significantly elevated levels of FCM on the first, second and third day after the introduction. For horses introduced together with an integration horse FCM were already significantly higher on the day of the introduction, indicating a stressful event before the introduction itself. In contrast, FCM levels were always very low when using the mixed technique.
In sum, horses have the ability to deal with conflict when they are introduced to new group members. The introduction event itself appears not to be as stressful as previously assumed. Standing together with an “integration horse” on a separate paddock and not being able to integrate immediately into a new group appears to be stressful for the newcomer. Based on the findings of our study we suggest to introduce new horses in group management together with a new group mate, a so-called “integration horse”. This would reduce the number of total social interactions as well as the aggression level. While this technique may be stressful for the newcomer, it lowers aggressive behaviour between the introduced horse and the group members and consequently reduces injury risks.

Abstract: In dieser Untersuchung sollte überprüft werden, ob die für die Stressmessung beim Menschen angewandte „Chronopsychobiologische Regulationsdiagnostik“ (CRD) mithilfe des smardwatch®-Messsystems auch beim Pferd psychische Belastungssituationen aussagekräftig widerspiegelt. Menschen können anhand variierender physiologischer und ethologischer Reaktionen auf Stressoren aus der Umwelt in Stresstypen eingeteilt werden. Auch Pferde reagieren in identischen Situationen ganz unterschiedlich. Es stellte sich daher die Frage, ob Pferde ebenfalls bestimmten Stresstypen zugeordnet werden können.
Mit 26 dreijährigen Hengsten des Landgestüts Celle wurde ein Verhaltenstest durchgeführt. Auf der Grundlage wissenschaftlich dokumentierter und modifizierter Testsituationen (Open-Field-Test, Novel-Object-Test, Startling-Test Objekt/Geräusch, Mensch-Pferd-Interaktion, Ressourcenkontrolle) wurde das Erkundungs- und Fluchtverhalten untersucht (Goslar 2011). Während dieser Belastungssituationen wurden mit dem smardwatch®-Messgerät die elektrophysiologischen Parameter Hautwiderstand, Hautpotential und Muskelaktivität erfasst. Durch eine Zeitreihenmessung dieser physiologischen Parameter konnten die Reaktionen des übergeordneten Reglers (vegetatives Nervensystem) dargestellt werden. Der Hautwiderstand spiegelt die vegetativ-emotionalen Reaktionen wider, das Hautpotential die vegetativ-nervalen und somit laut Balzer (2009) die kognitiven Verarbeitungsweisen, und durch das Elektromyogramm werden die typischen muskulär-motorischen Reaktionen aufgezeichnet. Die vorliegenden Datenzeitreihen wurden mit Hilfe einer biorhythmometrischen Zeitreihenanalyse nach Balzer und Hecht (Hecht 2001, Balzer 2009) ausgewertet. Als Ergebnis wurden chronobiologische Regulationszustände definiert, die dann gemäß dem „Periodischen System der Regulationszustände“ (PSR) (Balzer 2000) eine Beurteilung der psychischen Belastungssituation erlauben. Zur Bestimmung von Stresstypen wurde die Untersuchung nach dem Reiz-Reaktions-Prinzip in drei Phasen unterteilt: Ruhephase, Stressreizphase und Stressverarbeitungsphase. Der Verhaltenstest wurde nach einer Woche zur gleichen Tageszeit mit jedem Hengst einmal wiederholt. Als physiologische Vergleichsparameter wurden vor, während und nach der Belastung insgesamt zehn Speichelproben zur Cortisolbestimmung entnommen.
Die smardwatch®-Messtechnik ist prinzipiell geeignet die physiologischen Parameter Hautwiderstand, Hautpotential und Muskelaktivität auf der Haut des Pferdes aufzunehmen. Durch die biorhythmometrische Zeitreihenanalyse konnten chronobiologische Regulationszustände definiert werden.
Der Verhaltenstest führte bei allen Pferden zu einem signifikanten Anstieg der Cortisolwerte. Ein ebenfalls statistisch gesicherter Unterschied der aktivierten Cortisolwerte zwischen der ersten und zweiten Testwoche zeigt eine geringere Stressreaktion und weist auf einen Lerneffekt der Pferde hin:
Entsprechend der relativen Instabilitäten der Regulation der Körperfunktionen in Messphase 2 und 3 konnten die Pferde in vier Stressregulationstypen (nach Balzer u. Hecht 1996) eingeteilt werden: Stressbeherrscher, -bewältiger und kompensierer sowie Stressnichtbewältiger.
Anhand der Aktivierung von Sympathikus und Parasympathikus konnten zusätzlich vier verschiedene Vegetative Stresstypen unterschieden werden: Sympathikotoniker, Amphotoniker, Indifferenter Typ und Vagotoniker.
Die Cortisolreaktion der Hengste war individuell sehr unterschiedlich. Um die maximalen Anstiege vergleichen zu können, wurde der Trend der jeweiligen Cortisolverlaufskurve bestimmt. Anhand der Trend-korrigierten Cortisolkurven konnten einerseits die relativen Maxima der Cortisolreaktion besser verglichen werden, anderseits konnten die Pferde mittels des unterschiedlichen Trendes in drei Cortisolverlaufsgruppen eingeteilt werden: Tendenz fallend, gleichbleibend oder steigend.
Ein statistischer Nachweis für eine Abhängigkeit zwischen den verschiedenen Stresstypenklassifizierungen konnte anhand der geringen Stichprobenzahl nicht erbracht werden. Jedoch lassen sich Pferde, wie Menschen, unterschiedlichen Stresstypen zuordnen. Schwierig bleibt aber die objektive Beurteilung von Befindlichkeiten, da Empfindungen und Gefühle nur subjektiv wahrnehmbare Qualitäten sind, die von einer Reihe innerer und äußerer Faktoren abhängen. Die Verhaltenszuordnung emotionaler Zustände durch die „Chronopsychobiologische Regulationsdiagnostik“ (CRD) kann nicht ohne die Basis weiterer vergleichender Studien vom Mensch auf das Pferd übertragen werden.
Die CRD-Methode könnte allerdings einen interdisziplinären Ansatz ermöglichen und zukünftig neben den klassischen deskriptiven Verhaltensbeobachtungen bei der Beurteilung von Haltungs- und Umgangssituationen von Tieren wertvolle Aufschlüsse über die Fähigkeit zur Stressbewältigung und deren Konsequenzen für das Wohlbefinden der Tiere geben.
In this study we examined, whether the “Chronopsychobiological regulation diagnosis” (CRD) with the smardwatch®-system which is used to assess specific strain in humans, is also able to reflect convincingly specific strain in horses. Humans can be categorized into so-called stress types, because they react differently in physiology and behaviour to environmental stimuli. Concerning horses, it is also known that individuals react differently in identical situations. The question to be answered is, if it is possible to categorize horses into certain stress types as well.
We carried out a behavioural test with 26 three-year-old stallions of the State Stud of Celle, involving different test situations. On the basis of test situations, well known in scientific literature including slight modifications (open-field-test, novel-object-test, startling-test object/sound, human-horse-interaction, resource control) the explorative and flight behaviour of horses were examined (Goslar 2011). During these situations of strain the system smardwatch® measured the electro-physiological parameters skin resistance, skin potential and muscle activity. With time series analyses of these physiological parameters the reactions of the vegetative nervous system as superior control could be shown. The skin resistance reflects the vegetative-emotional, the skin potential the vegetative-nervous hence according to Balzer (2009) cognitive reactions and the electromyogram shows the motorized reactions. The time series of measured data was analyzed using the biorhythmometrical time series analysis of Balzer and Hecht (Hecht 2001, Balzer 2009). As a result of this, typical states of chronobiological regulation were defined. With the help of the “periodic system of regulatory states” (PSR) (Balzer 2000) these led to a classification of mental stress situations. To categorize horses into stress types the study was devided into three phases according to the stimulus-response principle: phase 1 (relaxing), phase 2 (situation of strain) and phase 3 (stimulus processing phase). The behavioural test was repeated once with each stallion exactly one week later. In addition ten samples of saliva were taken before, during and after the situations of strain from which we determined the cortisol concentration to be compared with the chronopsychobiological parameters.
The smardwatch®-measurement technology is fundamentally suitable to measure the physiological parameters skin resistance, skin potential and electrical muscle activity of horses. Using the biorhythmometrical time series analysis, chronobiological regulatory states could be defined for horses as well.
For each horse the cortisol value increased significantly during the behavioural test. A remarkable difference of the cortisol values assessed in the first and second test, indicates a learning effect: The behavioural test led to a significantly lower stress reaction in the second week.
Depending on the proportions of unstable regulation processes during and after the situations of strain, four types of regulation (Balzer u. Hecht 1996) can be defined: the Control-, Cope-, Compensate- and Non-cope-type.
Using the activation of the sympathetic and parasympathetic nervous system four vegetative stress types could be determined: Sympathicotonic, Amphotonic, Indifferent Type and Vagotonic.
The stress reaction of the stallions in terms of cortisol level was quite individual. To compare the maximum increase the trend of each cortisol trajectory was analyzed. On the one hand correcting for the trend allowed a comparison of relative maxima, on the other hand the horses could be categorized into groups, according to the evolution of their cortisol level (cortisol progression groups): Tendency falling, stable or rising.
The number of samples was not considered to be sufficient in order to statistically assess some dependence among the different classifications of stress types. But to our point of view horses can be assigned to different stress types like humans. But it’s still difficult to judge the emotions of animals, because emotions are subjectively perceptible qualities dependent on many internal and external factors. The assignment of emotional behaviour to the chronopsychobiological regulation diagnosis (CRD) can’t be transferred from human to horse without further studies.
However, the CRD method could enable an interdisciplinary approach. Besides classic descriptive observations of behavior, the CRD could give further information about the coping capacity and the consequences for animal welfare in the assessment of stressful situations.