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Vetvik, H., Grewal, H. M. S., Haugen, I. L., Åhrén, C., & Haneberg, B. (1998). Mucosal antibodies can be measured in air-dried samples of saliva and feces. Journal of Immunological Methods, 215(1–2), 163–172.
Abstract: IgA antibodies reflecting airways or intestinal mucosal immune responses can be found in saliva and feces, respectively, and IgG antibodies reflecting serum antibodies can be found in saliva. In this study, antibodies were detected in samples of saliva and feces which had been air-dried at room temperature (+20°C) or +37°C, and stored at these temperatures for up to 6 months. In saliva the antibody levels increased, while the antibodies in feces decreased upon storage. The individual IgA antibody concentrations which were adjusted by using the ratios of specific IgA/total IgA were relatively stable in both saliva and feces, and correlated with corresponding antibody levels in samples which had been stored at -20°C. The results indicate that air-dried saliva and feces can be used for semiquantitative measurements of mucosal antibodies, even after prolonged storage at high temperatures and lack of refrigeration.
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Sheriff, M. J., Dantzer, B., Delehanty, B., Palme, R., & Boonstra, R. (2011). Measuring stress in wildlife: techniques for quantifying glucocorticoids. Oecologia, 166(4), 869–887.
Abstract: Stress responses play a key role in allowing animals to cope with change and challenge in the face of both environmental certainty and uncertainty. Measurement of glucocorticoid levels, key elements in the neuroendocrine stress axis, can give insight into an animal’s well-being and can aid understanding ecological and evolutionary processes as well as conservation and management issues. We give an overview of the four main biological samples that have been utilized [blood, saliva, excreta (feces and urine), and integumentary structures (hair and feathers)], their advantages and disadvantages for use with wildlife, and some of the background and pitfalls that users must consider in interpreting their results. The matrix of choice will depend on the nature of the study and of the species, on whether one is examining the impact of acute versus chronic stressors, and on the degree of invasiveness that is possible or desirable. In some cases, more than one matrix can be measured to achieve the same ends. All require a significant degree of expertise, sometimes in obtaining the sample and always in extracting and analyzing the glucocorticoid or its metabolites. Glucocorticoid measurement is proving to be a powerful integrator of environmental stressors and of an animal’s condition.
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Mejdell, C. M., Buvik, T., Jørgensen, G. H. M., & Bøe, K. E. (2016). Horses can learn to use symbols to communicate their preferences. Appl. Anim. Behav. Sci., 184, 66–73.
Abstract: Abstract This paper describes a method in which horses learn to communicate by touching different neutral visual symbols, in order to tell the handler whether they want to have a blanket on or not. Horses were trained for 10–15 min per day, following a training program comprising ten steps in a strategic order. Reward based operant conditioning was used to teach horses to approach and touch a board, and to understand the meaning of three different symbols. Heat and cold challenges were performed to help learning and to check level of understanding. At certain stages, a learning criterion of correct responses for 8–14 successive trials had to be achieved before proceeding. After introducing the free choice situation, on average at training day 11, the horse could choose between a “no change” symbol and the symbol for either “blanket on” or “blanket off” depending on whether the horse already wore a blanket or not. A cut off point for performance or non-performance was set to day 14, and 23/23 horses successfully learned the task within this limit. Horses of warm-blood type needed fewer training days to reach criterion than cold-bloods (P < 0.05). Horses were then tested under differing weather conditions. Results show that choices made, i.e. the symbol touched, was not random but dependent on weather. Horses chose to stay without a blanket in nice weather, and they chose to have a blanket on when the weather was wet, windy and cold (χ2 = 36.67, P < 0.005). This indicates that horses both had an understanding of the consequence of their choice on own thermal comfort, and that they successfully had learned to communicate their preference by using the symbols. The method represents a novel tool for studying preferences in horses.
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Schuetz, A., Farmer, K., & Krueger, K. (2017). Social learning across species: horses (Equus caballus) learn from humans by observation. Anim. Cogn., 20(3), 567–573.
Abstract: This study examines whether horses can learn by observing humans, given that they identify individual humans and orientate on the focus of human attention. We tested 24 horses aged between 3 and 12. Twelve horses were tested on whether they would learn to open a feeding apparatus by observing a familiar person. The other 12 were controls and received exactly the same experimental procedure, but without a demonstration of how to operate the apparatus. More horses from the group with demonstration (8/12) reached the learning criterion of opening the feeder twenty times consecutively than horses from the control group (2/12), and younger horses seemed to reach the criterion more quickly. Horses not reaching the learning criteria approached the human experimenters more often than those that did. The results demonstrate that horses learn socially across species, in this case from humans.
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Karenina, K., Giljov, A., Ingram, J., Rowntree, V. J., & Malashichev, Y. (2017). Lateralization of mother�infant interactions in a diverse range of mammal species. Nat Ecol Evol, 1, 0030 Ep -.
Abstract: Left-cradling bias is a distinctive feature of maternal behaviour in humans and great apes, but its evolutionary origin remains unknown. In 11 species of marine and terrestrial mammal, we demonstrate consistent patterns of lateralization in mother�infant interactions, indicating right hemisphere dominance for social processing. In providing clear evidence that lateralized positioning is beneficial in mother�infant interactions, our results illustrate a significant impact of lateralization on individual fitness.
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M, E., östl,., Messmann, S., Bagu, E., Robia, C., & Palme, R. (1999). Measurement of Glucocorticoid Metabolite Concentrations in Faeces of Domestic Livestock. J. Vet. Med. A, 46(10), 621–631.
Abstract: After 14C-labelled cortisol infusion in ponies and pigs, faecal samples were collected. Extraction of 0.5 g faeces with 5 ml 80–90 % methanol yielded the highest radioactivity in the supernatant. Most of the metabolites were ether soluble. After high performance liquid chromatography (HPLC), the presence of immunoreactive metabolites was demonstrated by measuring each HPLC fraction using enzyme immunoassays for cortisol, corticosterone and 11-oxoaetiocholanolone. Only the assay for 11-oxoaetiocholanolone revealed peaks with co-eluting radioactivity. For biological validation of the test system, adrenocorticotrophic hormone (ACTH) and dexamethasone were injected intravenously successively in both species (n = 6). Cortisol concentration in blood and the 11-oxoaetiocholanolone immunoreactive substances in faeces were determined. In horse faeces, basal values of 2.3–35.2 nmol/kg were measured. After ACTH administration, an increase (more than 200 % above basal values) of these metabolites was seen about 1 day after ACTH administration. After dexamethasone injection the levels decreased, reaching minimum concentrations 2 days after administration. In pigs, an increase in these metabolites was measured in only three animals after ACTH; dexamethasone did not cause a decrease. The stability of the samples after defecation was tested by storing samples from cows, horses and pigs at room temperature. It was shown that there was a significant increase in the concentration of measured cortisol metabolites in bovine, equine and porcine faeces after storage for 1 h, 4 h and 24 h, respectively. In frozen samples this effect was diminished after thawing samples at 40°C; thawing the samples at 95°C prevented an increase in immunoreactive substances.
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Palme, R., Touma, C., Arias, N., Dominchin, M. F., & Lepschy, M. (2012). Steroid extraction: Get the best out of faecal samples. Vet. Med. Austria, 100, 238–246.
Abstract: Faecal steroid hormone metabolites are becoming increasingly popular as parameters for reproductive functions and stress. Theextraction 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 artificial 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.
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Weber-Mzell, D., Kotanko, P., Hauer, A. C., Goriup, U., Haas, J., Lanner, N., et al. (2004). Gender, age and seasonal effects on IgA deficiency: a study of 7293 Caucasians. European Journal of Clinical Investigation, 34(3), 224–228.
Abstract: Background The frequency of serum IgA deficiency (SIgAD) differs between populations. We examined the prevalence of SIgAD in healthy Caucasians. Materials and methods Serum immunoglobulin A (SIgA) was measured in 7293 volunteers (2264 women, 5029 men) aged 30 ± 14·2 years (mean ± SD; range: 12–66). Serum immunoglobulin A and subnormal SIgA levels were defined by a SIgA level < 0·07 g L-1, and between 0·07 and 0·7 g L-1, respectively. Means were compared by analysis of variance (anova) and analysis of covariance (ancova); frequencies by the χ2 test. Results Fifteen subjects (0·21%; one woman, 14 men) had SIgAD. Subnormal SIgA levels were found in 155 persons (2·13%): 21 females (0·93% of the females) and 134 males (2·66% of the males; difference: 1·74%; 95% CI: 1·12–2·33%; P < 0·001). Males were more likely to have subnormal SIgA levels or SIgAD (odds ratio 3·09, 95% CI: 1·97–4·85). The prevalence of SIgAD and subnormal SIgA was lowest in winter (χ2 = 14·8; P = 0·002; 3 d.f.; and χ2 = 43·2; P < 0·001; 3 d.f., respectively). Serum immunoglobulin A concentrations were significantly higher during winter. Serum immunoglobulin A levels increased with age on average by 0·2 ± 0·06 g L-1 per decade of life (P < 0·001). Taking into account the influence of age, SIgA concentration was lower in females as compared with males. Conclusion The prevalence of SIgAD and subnormal SIgA levels is increased in males. There exists a significant influence of gender, age and seasons on SIgA levels.
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Pinho, G. M., Gonçalves da Silva, A., Hrbek, T., Venticinque, E. M., & Farias, I. P. (2014). Kinship and Social Behavior of Lowland Tapirs (Tapirus terrestris) in a Central Amazon Landscape. Plos One, 9(3), e92507.
Abstract: We tested the hypothesis that tapirs tolerate individuals from adjacent and overlapping home ranges if they are related. We obtained genetic data from fecal samples collected in the Balbina reservoir landscape, central Amazon. Samples were genotyped at 14 microsatellite loci, of which five produced high quality informative genotypes. Based on an analysis of 32 individuals, we inferred a single panmictic population with high levels of heterozygosity. Kinship analysis identified 10 pairs of full siblings or parent-offspring, 10 pairs of half siblings and 25 unrelated pairs. In 10 cases, the related individuals were situated on opposite margins of the reservoir, suggesting that tapirs are capable of crossing the main river, even after damming. The polygamous model was the most likely mating system for Tapirus terrestris. Moran's I index of allele sharing between pairs of individuals geographically close (<3 km) was similar to that observed between individual pairs at larger distances (>3 km). Confirming this result, the related individuals were not geographically closer than unrelated ones (W = 188.5; p = 0.339). Thus, we found no evidence of a preference for being close to relatives and observed a tendency for dispersal. The small importance of relatedness in determining spatial distribution of individuals is unusual in mammals, but not unheard of. Finally, non-invasive sampling allowed efficient access to the genetic data, despite the warm and humid climate of the Amazon, which accelerates DNA degradation.
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Harewood, E. J., & McGowan, C. M. (2005). Behavioral and physiological responses to stabling in naive horses. J. Equine Vet. Sci., 25(4), 164–170.
Abstract: The purpose of this study was to investigate the response of horses to confinement and isolation in a stable (indoor individual housing) for the first time using behavioral indices, heart rate, and salivary cortisol concentration. Six naive 2-year-old Australian Stock Horse fillies were examined at 4-hour intervals over 24 hours in an outdoor group paddock followed by 24 hours in indoor individual housing. Behavioral observations and scores and heart rates were recorded and saliva samples were taken at each interval. During stabling, all horses became agitated and demonstrated increased vocalization and movement. Behavioral scores were significantly higher in the indoor individual housing (P < .001). No significant difference in heart rates between the two environments was detected. Mean salivary cortisol did not increase significantly (2 ng/mL ± 1.4 ng/mL in outdoor group paddock vs 2.5 mL ± 1.2 ng/mL in indoor individual housing). No diurnal rhythm in salivary cortisol was evident in either the outdoor group paddock or indoor individual housing. The results of this study highlight that a combination of behavioral and physiological measures allow better understanding of stress, where one measurement may be misleading. First time stabling of horses elicited marked behavioral responses indicative of stress that were not reflected in increased heart rates or salivary cortisol concentrations. The lack of a diurnal cortisol rhythm and the comparatively high basal cortisol concentrations found in the outdoor group paddock environment may imply that the fillies were already stressed; therefore, stabling did not cause further aberrations detectable by salivary cortisol analysis.
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