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Hothersall, B., Harris, P., Sörtoft, L., & Nicol, C. J. (2010). Discrimination between conspecific odour samples in the horse (Equus caballus). Appl. Anim. Behav. Sci., 126(1-2), 37–44.
Abstract: Behavioural observations suggest that smell is important in social discriminations between horses but balanced studies of this capacity are lacking. We used a habituation-discrimination procedure to investigate the ability of horses to distinguish between pairs of odour samples from different individuals. In Study 1, separate tests were conducted for urine, faeces or fleece fabric previously rubbed on the coat (to pick up body odour samples (BOS)) and donor pairs differed in sex, and age. 10 pregnant mares each underwent three tests, one per sample type. A test consisted of three successive 2-min presentations of a sample from Individual A with a simultaneous presentation of a sample from Individual B during the final presentation. Doubly repeated measures ANOVA indicated a main effect of sample type on investigative response (df = 2, f = 7.98, P = 0.004): durations were longer for BOS than for urine or faeces but habituation across trials was most consistent for urine. In the final presentation, mares demonstrated discrimination by investigating the novel urine sample (B) more than the repeated sample (novel: median 8.0 s, IQR = 10; repeated: median 2.5 s, IQR = 6; z = -2.558, P = 0.008). In Study 2, urine samples from castrated male donors were used and neither mares nor their 4-month-old foals discriminated between samples from different individuals in the final presentation. The findings suggest that urine odour may contain some information that horses can use to discriminate between conspecifics. This may be limited to the level of broad categories such as sex or reproductive status; further investigation is needed to reveal what functional information can be transmitted and what compounds are involved.
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Lamoot, I., Callebaut, J., Degezelle, T., Demeulenaere, E., Laquiere, J., Vandenberghe, C., et al. (2004). Eliminative behaviour of free-ranging horses: do they show latrine behaviour or do they defecate where they graze? Appl. Anim. Behav. Sci., 86(1-2), 105–121.
Abstract: In contrast to horses in pastures, it is thought that free-ranging horses do not perform latrine behaviour, i.e. a behavioural pattern whereby the animals graze and defecate in separate areas. However, few studies deal with this particular subject, reporting contrasting conclusions. We hypothesize that horses free-ranging in large heterogeneous areas do not perform latrine behaviour. Thus, we believe that grazing and elimination behaviour are spatially related: where horses graze, they will also defecate. Behavioural data were collected from Konik horses, Haflinger horses, Shetland ponies and donkeys, grazing in different nature reserves (54-80 ha). Data for the different equids were analyzed separately, as well as data for mares and stallions (Konik and donkey stallions only). We investigated the proportion of the number of defecations/urinations while grazing on the total number of defecations/urinations; furthermore, we searched for the sequence of behaviours representing latrine behaviour in the strict sense. Additionally, we analyzed the correlation between grazing behaviour and eliminative behaviour on both vegetation type level and patch level. All the female equids often continued grazing while defecating. During urination, grazing ceases in the majority of instances. Cases where a mare terminated grazing in a certain vegetation type and sward height to eliminate in another vegetation type or in another sward height within the same vegetation type were rarely observed. On the vegetation type level as well as on the patch level, there was a highly significant (P<0.001) positive correlation between grazing time and number of eliminations (or eliminating time). The high values of the correlation coefficients (in case of the defecation variables r ranges between 0.553 and 0.955; in case of the urination variables r ranges between 0.370 and 0.839) illustrate that the spatial distribution of the eliminative behaviour can be explained to a high degree by the spatial distribution of the grazing behaviour. Results in the case of the stallions are preliminary, but indicate the same pattern. Horses, free-ranging in large heterogeneous areas, do not perform latrine behaviour, but defecate where they graze. Possibly, animal density is of major importance to explain this behavioural difference with horses in pastures. We suggest that also spatial vegetation heterogeneity and plant productivity of the grazed area, as well as parasite status of the grazing animals could play a role.
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Palme, R. (2012). Monitoring stress hormone metabolites as a useful, non-invasive tool for welfare assessment in farm animals. Animal Welfare, 21(3), 331–337.
Abstract: A multitude of endocrine mechanisms are involved in coping with challenges. Glucocorticoids, secreted by the adrenal glands, are in the front line of the battle to overcome stressful situations. They are usually measured in plasma samples as parameters of adrenal activity and thus of disturbance. Unfortunately, collecting blood samples itself can disturb an animal. Thus, non-invasive methods for the determination of glucocorticoids or their metabolites have become increasingly popular. The pros and cons of various non-invasive sample materials (saliva, excreta, milk, hair/feathers and eggs) for glucocorticoid determination are given. Above all, faecal samples offer the
advantage that they can be collected easily. In faecal samples, circulating hormone levels are integrated over a certain period of time and represent the cumulative secretion of hormones. Thus, the levels are less affected by short fluctuations or the pulse-like nature of hormone secretion. However, using this technique to assess an animal’s adrenocortical activity is not especially simple. Whether frequent sampling is necessary or single samples will suffice depends upon the study’s aim (whether one is examining the impact of acute or chronic stressors). Background knowledge of the metabolism and excretion of cortisol/corticosterone metabolites is required and a careful validation for each species and sex investigated is obligatory. The present review also addresses analytical issues regarding sample storage, extraction procedures and immunoassays and includes a comprehensive list of published studies (up to 2011) describing the use of such methods in farmed animals. Applied properly, non-invasive techniques to monitor glucocorticoid metabolites in faecal samples of various species are a useful tool for welfare assessment, especially as they are easily applied at farm or group level.
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Palme, R., Fischer, P., Schildorfer, H., & Ismail, M. N. (1996). Excretion of infused 14C-steroid hormones via faeces and urine in domestic livestock. Animal Reproduction Science, 43(1), 43–63.
Abstract: The aim of this comparative study was to gain more information about the excretion of steroid hormones in farm animals. This should help to establish or improve non-invasive steroid monitoring procedures, especially in zoo and wildlife animals. Over a period of 4 h the 14C-steroid hormones (3.7 MBq) progesterone (three females), testosterone (three males), cortisol and oestrone (two males, two females) were infused intravenously in sheep, ponies and pigs. Faeces were collected immediately after defecation. Urine was sampled via a permanent catheter in females and after spontaneous urination in males. A total of 88 +/- 10% (mean +/- SD) of the administered radioactivity was recovered. Considerable interspecies differences were measured both in the amounts of steroid metabolites excreted via faeces or urine and the time course of excretion. Progesterone and oestrone in ewes, and progesterone in mares were excreted mainly in the faeces (over 75%). The primary route of excretion of all other 14C-steroids was via the urine but to a different extent. In general, sheep showed the highest degree of faecal excretion and pigs the least. The highest radioactivity in urine (per mmol creatinine) was observed during the infusion or in one of the next two samples thereafter, whereas in faeces it was measured about 12 h (sheep), 24 h (ponies) or 48 h (pigs) after the end of the infusion. Thereafter the radioactivity declined and reached background levels within 2-3 weeks. In faeces, steroid metabolites were present mainly in an unconjugated form, but in blood and urine as conjugates. Mean retention time of faecal radioactivity suggested that the passage rate of digesta (duodenum to rectum) played an important role in the time course of the excretion of steroids. The information derived from this investigation could improve the precision of sampling as well as the extraction of steroids from the faeces. Furthermore, the study demonstrates that it should be possible to establish methods for measuring faecal androgen and cortisol metabolites for assessing male reproductive endocrinology and stress in animals.
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Palme, R., Touma, C., Arias, N., Dominchin, M.N., & Lepschy, M. (2013). Steroid extraction: Get the best out of faecal samples. Wien Tierärztl Monat – Vet Med Austria, 100, 238–246.
Abstract: Faecal steroid hormone metabolites are becoming increasingly popular as parameters for reproductive functions and stress. The extraction of the steroids from the faecal matrix represents the initial step before quantification can be performed. The steroid metabolites present in the faecal matrix are of varying polarity and composition, so selection of a proper extraction procedure is essential. There have been some studies to address this complex but often neglected point. Radiolabelled steroids (e.g. cortisol or progesterone) have frequently been added to faecal samples to estimate the efficiency of the extraction procedures used. However, native, unmetabolized steroids are normally not present in the faeces and therefore the results are artifi- cial and do not accurately reflect the actual recoveries of the substances of interest. In this respect, recovery experiments based on faecal samples from radiometabolism studies are more informative. In these samples, the metabolite content accurately reflects the mixture of metabolites present in the given species. As a result, it is possible to evaluate different extraction methods for use with faecal samples. We present studies on sheep, horses, pigs, hares and dogs that utilized samples containing naturally metabolized, 14C-labelled steroids. We recommend extracting faecal steroids by simply suspending the faeces in a high percentage of a primary alcohol (for glucocorticoid metabolites 80% aqueous methanol proved best suited for virtually all mammalian species tested so far). Not only does the procedure significantly increase the total amount of recovered radioactivity, it also increases the percentage of unconjugated metabolites, which are more likely to be recognized by the antibodies used in various immunoassays. The advantages of this extraction procedure are clear: it is very easy to use (no evaporation step is needed), it yields high recoveries and variation based on the extraction procedure is reduced to a minimum.
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Pfammatter, M., Huwiler, S., Kägi, F., Kopp, C., Krüger, K., & Herholz, C. (2017). Leistung und Stresslevel bei Maultieren während eines fünftägigen Gotthardtrecks [Performance et niveau de stress chez les mulets durant un trek de cinq jours sur le Gothard][Muli: prestazione e livello di stress durante una traversata del Gottardo di 5 giorni][Performance and stress level in mules during a five days Gotthard trek]. Agrarforschung Schweiz, 8((7-8)), 276–283.
Abstract: Während einer fünftägigen Gotthardüberquerung im Sommer 2016 haben drei Maultiere als Tragtiere mit einer Gepäcklast von je 80 kg rund 94,46 Kilometer und 3�364 Höhenmeter bewältigt. Die Leistungsanforderung wurde anhand der Erholungswerte der Vitalparameter Herzfrequenz, Atemfrequenz und Körpertemperatur, sowie durch kontinuierliche Herzfrequenzmessung während der Belastung evaluiert. Die Bestimmung der Glucokortikoid- Metabolit Konzentration im Kot diente zur Einschätzung des Stresslevels der drei Maultiere. Die Erholungswerte der Herzfrequenzen der drei Maultiere lagen während allen Trekkingetappen in einem Bereich, der nicht auf eine Leistungsüberforderung schliessen liess. Anhand der kontinuierlichen Herzfrequenzaufzeichnung bei einem der Maultiere konnte gezeigt werden, dass die physische Leistungsanforderung im Ausdauerbereich lag. Wie als normale physiologische Reaktion des Körpers nach einer fünftägigen körperlichen Belastung erwartet, stieg Stresslevel gemessen an den Glucokortikoid- Metaboliten im Kot gegen Ende des Trecks bei allen Maultieren an. In der vorliegenden Studie konnte gezeigt werden, dass die Maultiere während des Gotthardtrecks ausdauernd belastbar waren, ohne durch die Anstrengung beeinträchtigt zu sein, die schon historisch von Maultieren abverlangt wurde.[
Durant une traversée du Gothard de cinq jours pendant l�été 2016, trois mulets ont été employés comme animaux de bât pour porter une charge de 80 kg sur 94,46 km, avec un dénivelé de 3364 m. Les exigences de performance ont été évaluées à partir des valeurs de récupération des paramètres vitaux (fréquence cardiaque, fréquence respiratoire et température corporelle) et de la fréquence cardiaque durant l�effort. La concentration en métabolites glucocorticoïdes dans le crottin a permis d�estimer le niveau de stress des trois animaux. Durant toute la durée du trek, les valeurs de récupération de la fréquence cardiaque des trois mulets étaient comprises dans un intervalle n�indiquant aucun effort excessif. La mesure constante des pulsations cardiaques a montré que l�intensité des performances physiques de ces animaux ne sortait pas de leur zone d�endurance. Comme on pouvait s�y attendre, le niveau de stress mesuré par le biais des métabolites glucocorticoïdes a augmenté chez tous les mulets à la fin du trek, réaction physiologique normale du corps après un effort de cinq jours. La présente étude a montré que durant un trek sur le Gothard, les mulets ont fait preuve d�endurance et de résistance, sans altération due à l�effort que l�on exige traditionnellement de leur part.][Nell�estate del 2016 tre muli impiegati come animali da soma hanno attraversato il Gottardo portando un carico di 80 kg ciascuno e percorrendo in cinque giorni un percorso di circa 94,46 chilometri con un dislivello di 3364 metri. La prestazione è stata valutata sulla base dei valori di recupero dei parametri vitali (frequenza cardiaca, frequenza respiratoria, temperatura corporea) e misurando costantemente la frequenza cardiaca sotto sforzo. Il livello di stress dei tre muli è stato valutato in funzione della concentrazione di metaboliti glicocorticoidi riscontrata nelle feci. In tutte le tappe del trekking i valori di recupero della frequenza cardiaca registrati nei muli rientravano in un intervallo che non lasciava supporre un sovraccarico. Sulla base della registrazione costante della frequenza cardiaca di uno dei tre muli è stato possibile dimostrare che l�intensità dello sforzo fisico non superava il loro intervallo di resistenza. Come previsto, verso la fine del trekking la concentrazione di metaboliti glicocorticoidi nelle feci di tutti e tre i muli indicava un aumento del livello di stress; si tratta di una reazione fisiologica normale dopo uno sforzo fisico di cinque giorni. Questo studio ha dimostrato che lo sforzo sopportato dai muli per portare un carico attraverso il Gottardo, un compito da secoli richiesto a questi animali, rientrava nel loro intervallo di resistenza e non ha avuto ripercussioni negative sul loro fisico.][In the summer of 2016, three pack mules, each carrying a load weighing 80kg, accompanied a 94.46km trek across the Gotthard Pass with a total altitude difference of 3,364m. The mules� performances were evaluated by measuring vital recovery parameters such as heart- and respiratory rates and body temperature, and by continuous heartrate monitoring during the trek. The stress levels of the animals were estimated by determining glucocorticoid metabolite levels in their faeces. Throughout the trekking days, recovery heartrates lay within a range which indicated that the animals were not being overworked. The continuous heartrate monitoring of one of the mules showed that its physical performance lay within the endurance zone. As expected, glucocorticoid metabolite levels were elevated in the faeces of all the mules � a normal physiological response after five consecutive days of exercise. The study shows the mules as being capable of performing at an endurance level during the Gotthard trek with no adverse affects on their health � a performance which was historically expected of the animals.]
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