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Author Carlsson, H.-E.; Lyberg, K.; Royo, F.; Hau, J. url  doi
openurl 
  Title Quantification of stress sensitive markers in single fecal samples do not accurately predict excretion of these in the pig Type Journal Article
  Year 2007 Publication Research in Veterinary Science Abbreviated Journal  
  Volume 82 Issue (down) 3 Pages 423-428  
  Keywords Cortisol; Immunoglobulin A; Stress; Pigs; Feces; Animal welfare  
  Abstract All feces produced during 24 h were collected from five pigs and cortisol and immunoreactive cortisol metabolites (CICM), and IgA were quantified. Within pigs, the concentrations of CICM and IgA varied extensively between random samples obtained from a single fecal dropping, and deviated in most cases significantly from the true concentration measured in total fecal output (CV 6.7–130%). The CICM and IgA contents varied considerably (CV 8.1–114%) within and between individual fecal droppings from the same pig compared to the total fecal excretion. In conclusion, single random samples could not be used to reliably quantify the total fecal concentration or excretion of CICM or IgA in pigs. Analyses of all feces collected during shorter periods than 24 h did not provide an accurate estimate of the daily excretion of CICM. Thus, the concentration of stress sensitive molecules in random single fecal samples as an indicator of animal welfare should be interpreted with prudence.  
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  ISSN 0034-5288 ISBN Medium  
  Area Expedition Conference  
  Notes Approved no  
  Call Number Equine Behaviour @ team @ Serial 5853  
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Author Rivera, E.; Benjamin, S.; Nielsen, B.; Shelle, J.; Zanella, A.J. url  doi
openurl 
  Title Behavioral and physiological responses of horses to initial training: the comparison between pastured versus stalled horses Type Journal Article
  Year 2002 Publication Applied Animal Behaviour Science Abbreviated Journal Appl. Anim. Behav. Sci.  
  Volume 78 Issue (down) 2-4 Pages 235-252  
  Keywords Horses; Housing; Training; Behavior; Stress; Cortisol; Welfare  
  Abstract Horses kept in stalls are deprived of opportunities for social interactions, and the performance of natural behaviors is limited. Inadequate environmental conditions may compromise behavioral development. Initial training is a complex process and it is likely that the responses of horses may be affected by housing conditions. Sixteen 2-year-old Arabian horses were kept on pasture (P) (n=8) or in individual stalls (S) (n=8). Twelve horses (six P and six S) were subjected to a standardized training procedure, carried out by two trainers in a round pen, and 4 horses (two P and two S) were introduced to the round pen but were not trained (C; control). On sample collection day 0, 7, 21 and 28, behavior observations were carried out, blood samples were drawn and heart rates were monitored. Total training time for the stalled horses was significantly higher than total time for the pastured horses (S: 26.4+/-1.5 min; P: 19.7+/-1.1; P=0.032). The stalled group required more time to habituate to the activities occurring from the start of training to mounting (S: 11.4+/-0.96; P: 7.3+/-0.75 min; P=0.007). Frequency of unwanted behavior was higher in the stalled horses (S: 8.0+/-2.0; P: 2.2+/-1.0; P=0.020). Pastured horses tended to have higher basal heart rates on day 0 (S: 74.7+/-4.8; P: 81.8+/-5.3 bpm; P=0.0771). While the physiological data failed to identify differences between housing groups, the behavioral data suggest that pasture-kept horses adapt more easily to training than stalled horses.  
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  Notes Approved no  
  Call Number refbase @ user @ Serial 717  
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Author Simpson, B.S. doi  openurl
  Title Neonatal foal handling Type Journal Article
  Year 2002 Publication Applied Animal Behaviour Science Abbreviated Journal Appl. Anim. Behav. Sci.  
  Volume 78 Issue (down) 2-4 Pages 303-317  
  Keywords Neonatal foal handling; Imprint training; Learning in foals; Heart rate; Serum cortisol concentrations  
  Abstract Recent interest has focused on the advantage of intensively handling young foals as a means of producing more tractable horses, accustomed to humans and receptive to training. To date, the effect of this intensive handling, dubbed “imprint training” in the popular literature, has not been tested. The present study compares seven foals handled intensively as neonates with eight untreated controls. The handling protocol started from 2-8 h after birth and continued daily for a total of 5 days. The protocol consisted of a series of stimuli and experiences that were each repeated until the foal no longer resisted or reacted negatively. Subsequently, foals were tested before weaning, at 4 months of age. Results indicated that handled foals (HF) ranked higher than control foals (CF) in subjective ratings of calmness (*P<0.0117) and friendliness (*P<0.0001) and in several specific handling tasks (venipuncture *P<0.0220; placing in stock *P<0.0128). Although, in approach tests all foals but one allowed approach of a person to 4 m, significantly more HF approached the person than CF (P<0.0080). In stimulus tests, foals were presented specific stimuli to which they had been tested as neonates. Two of eight CF were too unruly and dangerous to test. Of foals that could be tested, CF required significantly more time to hook-up a heart rate monitor (**P<0.0055). Split-plot analysis indicated that HF had lower heart rates to initial left-sided stimuli, presented first, than CF (*P<0.0421). In response to right-sided stimuli, heart rate scores of CF were not significantly different from HF (P<0.2259), suggesting reduced reactivity over time due to a learning effect. Behavioral responses to specific stimuli did not differ between CF and HF, suggesting that neonatal handling has a general rather than specific effect on subsequent behavior. Cortisol concentrations were measured before and after testing and the difference calculated. All foals had higher post-testing levels than pre-testing levels. There was a significant difference between HF and CF, indicating greater reactivity among the CF (*P<0.050). In general, the results indicated that foals handled as neonates were more tractable and less reactive. Specific neonatal handling tasks, such as sticking a finger up the foal's nose or patting the bottom of the foot, seemed to have no beneficial effect on related tasks such as passing a nasogastric tube or tapping with a farrier's hammer at 4 months of age. Mechanisms for the observed effect of neonatal handling require further investigation.  
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  Notes Approved no  
  Call Number Equine Behaviour @ team @ Serial 3623  
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Author Flauger, B.; Krueger, K.; Gerhards, H.; Möstl, E. url  doi
openurl 
  Title Simplified method to measure glucocorticoid metabolites in faeces of horses Type Journal Article
  Year 2010 Publication Veterinary Research Communications Abbreviated Journal Vet Res Comm  
  Volume 34 Issue (down) 2 Pages 185-195  
  Keywords ACTH challenge; enzyme immunoassay; stress behaviour; cortisol  
  Abstract Glucocorticoids or their metabolites can be measured in several body fluids or excreta, including plasma, saliva, urine and faeces. In recent years the measurement of glucocorticoid metabolites (GCMs) in faeces has gained increasing attention, because of its suitability for wild populations. In horses, however, the group-specific enzyme immunoassay described so far has a limited racticability due to its complex extraction procedure. Therefore, we tested the applicability of

other enzyme immunoassays for glucocorticoid metabolites. The present study clearly proved that an enzyme immunoassay (EIA) for 11-oxoetiocholanolone using 11-oxoetiocholanolone-17-CMO: BSA (3&#945;,11-oxo-A EIA) as antigen showed high amounts of immunoreactive substances. Therefore it was possible to use just a small amount of the supernatant of a methanolic suspension of faeces. The results

correlated well with the already described method for measuring GCMs in horse faeces, i.e. analysing the samples with an EIA after a two step clean up procedure of the samples (Merl et al. 2000). In addition, the 3&#945;,11-oxo-A EIA has the advantage of providing a bigger difference between baseline values and peak values after ACTH stimulation. The new assay increased the accuracy of the test,

lowered the expenses per sample, and storing samples at room temperature after collection was less critical than with other assays investigated in our study. This is a big advantage both in the field of wildlife management of equids and in the field of equestrian sports and it shows the importance of choosing an assay which is in good accordance with the metabolites excreted in a given species.
 
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  Notes Approved no  
  Call Number Equine Behaviour @ team @ Serial 5073  
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Author Schmidt, A.; Möstl, E.; Wehnert, C.; Aurich, J.; Müller, J.; Aurich, C. url  doi
openurl 
  Title Cortisol release and heart rate variability in horses during road transport Type Journal Article
  Year 2010 Publication Hormones and Behavior Abbreviated Journal Horm. Behav.  
  Volume 57 Issue (down) 2 Pages 209-215  
  Keywords Horse; Transport; Cortisol; Heart rate variability  
  Abstract Based on plasma cortisol concentrations it is widely accepted that transport is stressful to horses. So far, cortisol release during transport has not been evaluated in depth by non-invasive techniques such as analysis of salivary cortisol and faecal cortisol metabolites. Transport also causes changes in heart rate and heart rate variability (HRV). In this study, salivary cortisol, faecal cortisol metabolites, heart rate and HRV in horses transported by road for short (one and 3.5 h) and medium duration (8 h) were determined. With the onset of transport, salivary cortisol increased immediately but highest concentrations were measured towards the end of transport (4.1 ± 1.6, 4.5 ± 2.6, 6.5 ± 1.8 ng/ml in horses transported for one, 3.5 and 8 h, respectively). Faecal cortisol metabolite concentrations did not change during transport, but 1 day after transport for 3.5 and 8 h had increased significantly (p < 0.01), reflecting intestinal passage time. Compared to salivary cortisol, changes in faecal cortisol metabolites were less pronounced. Heart rate increased and beat-to-beat (RR) interval decreased (p < 0.05) with the onset of transport. Standard deviation of heart rate increased while root mean square of successive RR differences (RMSSD) decreased in horses transported for 3.5 (from 74 ± 5 to 45 ± 6 ms) and 8 h (from 89.7 ± 7 to 59 ± 7 ms), indicating a reduction in vagal tone. In conclusion, transport of horses over short and medium distances leads to increased cortisol release and changes in heart rate and HRV indicative of stress. The degree of these changes is related to the duration of transport. Salivary cortisol is a sensitive parameter to detect transient changes in cortisol release.  
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  Series Volume Series Issue Edition  
  ISSN 0018-506x ISBN Medium  
  Area Expedition Conference  
  Notes Approved no  
  Call Number Equine Behaviour @ team @ Serial 5387  
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Author Tiefenbacher, S.; Lee, B.; Meyer, J.S.; Spealman, R.D. url  doi
openurl 
  Title Noninvasive technique for the repeated sampling of salivary free cortisol in awake, unrestrained squirrel monkeys Type Journal Article
  Year 2003 Publication American Journal of Primatology Abbreviated Journal Am. J. Primatol.  
  Volume 60 Issue (down) 2 Pages 69-75  
  Keywords saliva; cortisol; squirrel monkey; sampling technique; Hpa  
  Abstract The use of noninvasive measures of hypothalamic-pituitary-adrenal (HPA) axis function is of growing interest among preclinical and clinical investigators. This report describes a method for the repeated assessment of salivary free cortisol in awake, unrestrained squirrel monkeys (Saimiri sciureus) based on a saliva sampling technique previously developed for rhesus monkeys. Individually housed adult male squirrel monkeys were trained to chew on dental rope attached to a pole, from which saliva was extracted by centrifugation and analyzed for cortisol by radioimmunoassay (RIA). Eight of nine monkeys readily acquired the task, reliably providing adequate saliva samples for the assay. Salivary free cortisol levels were examined in these subjects under basal conditions and in response to two types of neuroendocrine challenge. Levels of salivary free cortisol showed relatively low intra- and interindividual variability, with mean individual morning levels ranging between 17.1 and 37.9 µg/dl. Squirrel monkeys demonstrated a consistent daily rhythm in salivary free cortisol ranging from a high of 27.4 ± 5.2 µg/dl (mean ± SEM) at 12 P.M. to a low of 7.5 ± 1.6 µg/dl at 6 P.M.. Intravenous (IV) challenges with 1 µg/kg ACTH, or 10 and 50 µg/kg CRF resulted in significant increases in salivary free cortisol. The described sampling technique provides a reliable and sensitive means for repeated measurement of HPA activity in unrestrained, awake squirrel monkeys. In addition, our findings illustrate several features of HPA system rhythmicity and reactivity using salivary cortisol instead of blood plasma or serum. Am. J. Primatol. 60:69–75, 2003. © 2003 Wiley-Liss, Inc.  
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  Publisher Wiley Subscription Services, Inc., A Wiley Company Place of Publication Editor  
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  Series Editor Series Title Abbreviated Series Title  
  Series Volume Series Issue Edition  
  ISSN 1098-2345 ISBN Medium  
  Area Expedition Conference  
  Notes Approved no  
  Call Number Equine Behaviour @ team @ Serial 5573  
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Author Young, T.; Creighton, E.; Smith, T.; Hosie, C. url  doi
openurl 
  Title A novel scale of behavioural indicators of stress for use with domestic horses Type Journal Article
  Year 2012 Publication Applied Animal Behaviour Science Abbreviated Journal  
  Volume 140 Issue (down) 1–2 Pages 33-43  
  Keywords Horse; Behaviour scores; Cortisol; Saliva; Welfare assessment; Non-invasive  
  Abstract Behaviour scores (BS) offer non-invasive, objective and easy to use ways of assessing welfare in animals. Their development has, however, largely focused on behavioural reactions to stressful events (often induced), and little use of physiological measures has been made to underpin and validate the behavioural measures. This study aimed to develop a physiologically validated scale of behavioural indicators of stress for the purpose of welfare assessment in stabled domestic horses. To achieve this, behavioural and physiological data were collected from 32 horses that underwent routine husbandry procedures. Principal component analysis (PCA) of the behavioural and physiological data revealed three meaningful components that were used as the basis of the scale. Analysis of video clips of the horses’ responses to the husbandry procedures was undertaken by a panel of equestrian industry professionals using a free choice profiling (FCP) methodology. These results were added to the scale along with key definitions from relevant literature. Salivary cortisol levels were significantly correlated with the BS confirming the scale was meaningful and reflected physiological stress. The scale offers an easy to use ‘tool’ for rapid, reliable non-invasive welfare assessment in horses, and reduces the need for potentially invasive physiological measures.  
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  ISSN 0168-1591 ISBN Medium  
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  Notes Approved no  
  Call Number Equine Behaviour @ team @ Serial 5637  
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Author Cerasoli, F.; Podaliri Vulpiani, M.; Saluti, G.; Conte, A.; Ricci, M.; Savini, G.; D'Alterio, N. doi  openurl
  Title Assessment of Welfare in Groups of Horses with Different Management, Environments and Activities by Measuring Cortisol in Horsehair, Using Liquid Chromatography Coupled to Hybrid Orbitrap High-Resolution Mass Spectrometry Type
  Year 2022 Publication Animals Abbreviated Journal Animals  
  Volume 12 Issue (down) 14 Pages  
  Keywords cortisol; animal welfare; horse; Lc-Hrms/Ms  
  Abstract Horses have always been animals used for companionship, work, transportation, and performance purposes over the history of humanity; there are different ways of managing horses, but studies on how horse welfare is influenced by different activities and managements are scanty. Understanding how the management, the environment, and the different uses of horses can affect the level of stress and well-being is important not only for people associated with horses. Three groups of horses with different management, environments, and activities were selected: (1) stabled horses ridden frequently, (2) horses that perform public order service under the Italian state police, and (3) free-ranging horses. Cortisol analysis was carried out on horsehair samples using liquid chromatography coupled to hybrid orbitrap high-resolution mass spectrometry (LC-HRMS/MS), a laboratory technique used for the first time to quantify horsehair cortisol. The selection of horses to be included in the three groups was carried out by including only subjects with positive welfare assessment in accordance with the horse welfare assessment protocol (AWIN). These analyses demonstrated that the cortisol levels detected in the horsehair of free-ranging animals were significantly higher compared to those detected in stabled and working horses. These results may have been a consequence of complex environmental, managerial, and behavioral factors, which should be worth further investigation  
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  Language Summary Language Original Title  
  Series Editor Series Title Abbreviated Series Title Animals  
  Series Volume 12 Series Issue 14 Edition  
  ISSN 2076-2615 ISBN Medium  
  Area Expedition Conference  
  Notes Approved no  
  Call Number Equine Behaviour @ team @ Serial 6674  
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Author Gröschl, M.; Wagner, R.; Rauh, M.; Dörr, H.G. url  doi
openurl 
  Title Stability of salivary steroids: the influences of storage, food and dental care Type Journal Article
  Year 2001 Publication Steroids Abbreviated Journal  
  Volume 66 Issue (down) 10 Pages 737-741  
  Keywords Cortisol; 17OH-Progesterone; Progesterone; Saliva; Stability  
  Abstract We studied influences of dental care, food and storage on the reproducibility of salivary steroid levels. Cortisol (F), 17OH-progesterone (17OHP) and Progesterone (P) were measured using adapted commercial radioimmunoassays. Saliva samples of healthy adults (n = 15; m:8; f:7) were collected directly before and after dental care, and directly before and after breakfast with various foodstuffs. A second experiment investigated stability of steroids under different storage conditions. Four series of identical saliva portions (I: Native saliva; II: Centrifuged saliva; III: Saliva with trifluor acetate (TFA); IV: Saliva with 0.5% NaN3) were stored at room temperature and at 4°C for up to three weeks. To demonstrate influences of repeated thawing and re-freezing of saliva on steroid values, saliva samples (n = 15) were divided into identical portions. These portions were frozen and re-thawed up to 5 times before measurement. Neither dental care nor intake of bread or milk effected the reproducibility of F, 170HP, and P. Steroid levels decreased significantly in the course of three weeks under different storage conditions (P < 0.001). This decrease was clinically relevant from the second week onward, with exception of NaN3 treated samples. After repeated freezing and re-thawing 17OHP and P decreased slightly (about 5%). Only F decreased significantly after the third thawing (P < 0.001). The results show the usefulness of standardized handling of saliva samples for improving reproducibility and reliability of salivary steroid measurements.  
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  ISSN 0039-128x ISBN Medium  
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  Notes Approved no  
  Call Number Equine Behaviour @ team @ Serial 5561  
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Author Dalmau, A.; Ferret, A.; Chacon, G.; Manteca, X. doi  openurl
  Title Seasonal Changes in Fecal Cortisol Metabolites in Pyrenean Chamois Type Journal Article
  Year 2007 Publication Journal of Wildlife Management Abbreviated Journal J Wildl Manag  
  Volume 71 Issue (down) 1 Pages 190-194  
  Keywords Cadí-Moixeró, Nature Reserve, chamois, cortisol metabolites, feces, hunting reserve, Pyrenees, Rupicapra pyrenaica pyrenaica, seasonal rhythm, stress  
  Abstract We studied seasonal changes in fecal cortisol metabolites (FCM), which have been widely used as indicators of stress, in a population of Pyrenean chamois (Rupicapra pyrenaica pyrenaica) in the Cad&#237; Range of northeastern Spain. We collected fecal samples from 2001 to 2003 in 3 particular locations with different altitudes and male or female presence, and we analyzed them for FCM and fecal nitrogen as an indicator of diet quality. We observed a clear seasonal pattern, with the highest FCM in winter, and we obtained correlations between FCM and monthly mean minimum temperatures and fecal nitrogen. We observed no effects of tourism presence, trophy hunting, or rut season on FCM. Analysis of cortisol metabolites in feces can be a good measure of winter stress in Pyrenean chamois.  
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  Notes Approved no  
  Call Number Equine Behaviour @ team @ Serial 4254  
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