Fecal Cortisol Concentrations Research Articles

Is the judgment bias test a good tool to assess the quality of horse management?

Animal welfare should include the possibility of animals experiencing positive emotions. Emotions influence the cognitive process, and judgment bias tests (JBTs) are employed in different species, to assess the optimistic or pessimistic expectation of an individual and its affective state. Only recently the JBTs have been applied to horses. This research aims to investigate the relationship between a spatial JBT and hypothalamic-pituitary-adrenal axis (HPA) chronic and acute activation in forty-one animals hosted in different kinds of environments: traditional stables (TS), natural boarding (NB), and ethological stable (ES). Fecal (FC) and horsehair (HC) cortisol concentrations were quantified for each subject through Radio-Immuno-Assay (RIA). Body condition score (BCS), as an indirect index of animal motivation towards food, and personality traits were measured to explore their possible influence on JBT results. Horses had to distinguish a positive position (P), where a bucket full of food was positioned, from a negative one (N), with an empty bucket. Then, 3 intermediate positions (Near Negative-NN, Near positive-NP, and Medium-M) with an empty bucket were presented to the subject one at a time. Only 20 subjects out of 41 completed the JBT and were included in the statistical analysis, and both BCS and P position, whether at the right or the left of the subject, seems to have influenced the inclusion rate. Only the ES group registered a significantly lower score in NN, suggesting a more optimistic affective state, whereas NB and TS did not significantly differ in their responses. Despite this, horses from NB recorded higher FC concentration than TS subjects during all the phases of the test, but lower HC levels, which could suggest a generally lower level of chronic stress but its interpretation presents several confounding factors. These results put into question whether JBT is indeed a good test to monitor the quality of the management, as it does not seem to reflect the chronic physiological state of the animals and could be influenced by a state of acute stress, caused by the test procedure. Due to these confounding factors, this procedure should be accompanied by other indicators. Finally, to include more animals and exclude possible biases, the structure of the JBT and the employment of food as a reward should be evaluated considering the peculiarities of the species and individual motivations.

Effects of different weaning times on the stress response and the intestinal microbiota composition of female forest musk deer (Moschus berezovskii) and their fawns.

The effects of mother-infant separation (i.e., weaning) on the physiology, psychology and nutrition of mammalian infants have attracted much attention. Forest musk deer (FMD) is a first-class protected species in China and listed endangered in the IUCN Red List. The captive breeding population is not only an important source for restocking of wild resources, but also a necessary way to supply the market with legal musk. So far, there is no scientific basis for the appropriate separation time of FMD females and their infants. Therefore, we used metagenome sequencing and enzyme-linked immunosorbent assays to study changes in the fecal cortisol concentration, as well as the intestinal microbiome composition and function of females and fawns at three different separation times, i.e., after 80 days, 90 days and 100 days. The results showed that the increment of the cortisol concentration in female FMD increased with increasing lactation time. The increment of cortisol concentration in infant FMD was highest in the 80 days weaning group, but there was no significant difference between the 90 days and the 100 days separation time. Based on the annotation results of COG, KEGG and CAZy databases, the abundance of different functions annotated by the intestinal microbiome of mothers and fawns of the 90 days weaning group changed slightly after separation. Based on the above results, the separation of mother and infant FMD is recommended after 90 days, i.e., the separation time that triggered the lowest rate of weaning stress and that supported a relatively stable gastro-intestinal physiology.

Evaluation of Stress Accompanying Immunocontraceptive Vaccination in Donkeys.

Simple SummaryThe overpopulation of donkeys is of concern worldwide. Some of the concerns associated with uncontrolled donkey populations are the destruction of habitat, competition for feed resources, and negative human–animal interactions. One of the most effective and humane solutions is the use of immunocontraception. Immunocontraception is the use of an animal’s immune system to prevent it from fertilizing offspring by the administration of a vaccine that targets a specific portion of the reproductive system. This study sought to measure and compare the amount of stress that is imposed when two different immunocontraceptives were administered to female donkeys. The results showed that physical exam parameters (temperature, pulse, and respiration) were not affected significantly but the measurement of stress hormones suggests an acute stress response after the first vaccination that was followed by a chronic stress response after a booster vaccination 35 days later, especially in donkeys that had reactions at the injection site. Further studies would be required to identify and reduce the stress associated with the immunocontraception vaccines that were used.The overpopulation of donkeys is recognized as a problem in many parts of the world. The main concerns with uncontrolled donkey populations are habitat degradation and competition for feed resources between donkeys and other species. One of the most effective and humane solutions is the use of immunocontraception. Therefore, this study sought to evaluate the stress imposed by the use of two formulations of a zona pellucida (ZP) vaccine, a recombinant (reZP) and a native porcine (pZP) vaccine, both formulated with a Freund’s adjuvant. The stress was objectively measured using fecal cortisol concentrations and physical examination parameters at fixed points before and after vaccination. We hypothesized that fewer changes in physical exam parameters and lower fecal cortisol concentrations would be stimulated in jennies treated with the reZP vaccine due to the selection of specific proteins. Twenty-five reproductively sound jennies were randomly assigned to reZP (n = 9), pZP (n = 8) or control (n = 8) groups. The vaccines were administered at five-week intervals. Physical exam parameters and body wall thickness of injection sites were recorded for each jenny for four days post-injections. Fecal samples were obtained every other day from day 0 (first vaccination) through day 6 and on days 35 to 41 after booster. Injection site reactions were common in all groups with the reZP and pZP groups being overrepresented. Lameness was observed in the pZP and reZP groups that were affected by injection site reactions and open abscesses. The present study showed an increase in fecal cortisol concentrations within 4 days after the first vaccination with ZP vaccines and, thereafter, a decrease in cortisol 35 days later after the second vaccination, especially in donkeys with open abscesses. Our results suggest that acute stress (increased cortisol) was induced after the first vaccination, and chronic stress (decreased cortisol) occurred thereafter in association with open abscesses. In conclusion, reZP and pZP formulated with Freund’s adjuvant induced local inflammatory reactions with a differential degree of acute and chronic stress in donkeys.

Annual dynamics between serum cortisol concentration and fecal cortisol concentration in a female captive tiger

Annual dynamics between serum cortisol concentration and fecal cortisol concentration in a female captive tiger

Variations in Behavioral and Physiological Traits in Yearling Tibetan Sheep (Ovis aries)

Simple SummaryTibetan sheep (Ovis aries) are raised in the wild by shepherds on the Tibetan Plateau and domesticated for a long period. Studying variations in the behavioral and physiological traits of sheep during the growing season is important for welfare and productivity. In this study, the behavior and physiology of a Tibetan sheep cohort were repeatedly evaluated in 2-, 5-, and 11-month-old sheep. The mean level of the risk-taking variable increased, while that of the vocalizations variable decreased. The exploration variable was stable over ontogeny. The novelty decreased, and the heart rate increased from 2 to 5 months, but both stabilized at 11 months. The fecal cortisol concentration (CORT) variable was stable at 2 and 5 months but decreased at 11 months of age. Stable correlations were reported for 2- and 5-month-olds and for behavioral variables and heart rate. However, some correlations emerged only at 11 months, whereas others disappeared over ontogeny. Moreover, CORT was independent of temperament and heart rate during the entire period.Temperament is a consistent behavioral difference among individuals over time or in different contexts. A comprehensive understanding of temperament and complex behavioral interactions enhances knowledge on animal evolution, welfare, and productivity. However, reports on the development of behavioral consistency over ontogeny are vague. Here, we tested the ontogeny of the temperament and physiological traits of Tibetan sheep (Ovis aries) in three crucial age stages. The mean level of the risk-taking variable increased, while that of the vocalizations variable decreased. The exploration variable was stable over ontogeny. The novelty decreased and the heart rate increased from the juvenile to the adolescent stage but stabilized at the adult stage. The fecal cortisol concentration (CORT) variable was stable at the juvenile and adolescent stages but decreased at the adult stage. Stable correlations were reported for the juvenile and adolescent stages and for the behavioral variables and heart rate. However, some correlations emerged only after maturation, whereas others disappeared over ontogeny. Moreover, CORT was independent of temperament and heart rate at different ages. These results demonstrate that age affects temperament and physiology and their correlations. Hence, developmental aspects should be incorporated into future temperament studies.

輸送距離の違いが黒毛和種肥育素牛の被毛と糞便中コルチゾル含有量および血液成分に及ぼす影響

輸送距離の違いが黒毛和種肥育素牛の被毛と糞便中コルチゾル含有量および血液成分に及ぼす影響

Effects of scent enrichment on behavioral and physiological indicators of stress in zoo primates.

Captive breeding is vital for primate conservation, with modern zoos serving a crucial role in breeding populations of threatened species and educating the general public. However, captive populations can experience welfare issues that may also undermine their reproductive success. To enhance the wellbeing of endangered zoo primates, we conducted a study to assess the effects of a new scent enrichment program on captive red-ruffed lemurs (Varecia rubra), black howler monkeys (Alouatta caraya), siamangs (Symphalangus syndactylus), lar gibbons (Hylobates lar) and orangutans (Pongo pygmaeus pygmaeus). We combined behavioral observations and fecal endocrinology analyses to evaluate the effects of a series of essential oils (benzoin, lavender, lemongrass) on five captive troops (N = 19) housed at Dudley Zoo & Castle and Twycross Zoo (UK). We recorded observations of natural species-specific and abnormal stress-related behaviors for 480 h using instantaneous scan sampling. We collected 189 fecal samples and measured the fecal cortisol concentrations using radioimmunoassay. We found a significant effect of the scent enrichment on behaviors, with red-ruffed lemurs and black howler monkeys reducing their social interactions, as well as red-ruffed lemurs and lar gibbons decreasing their stress-related behaviors after they were exposed to the series of essential oils. We also found that red-ruffed lemurs displayed a significant increase in fecal glucocorticoids following exposure to essential oils. Our contradictory findings suggest that the effects of this series of essential oils may change depending on the species-specific social lives and olfactory repertoires of primates. In conclusion, we cannot recommend using these essential oils widely with zoo primates without additional evaluation.

The effects of a training program using a phantom to accustom heifers to the automatic milking system

The introduction of heifers into the automatic milking system (AMS) can be associated with considerable stress for both animals and farm employees, as completely inexperienced heifers initially do not independently enter the unknown milking robot. This study investigated whether training heifers on an AMS phantom provides the possibility of preparing heifers for the following lactation at the AMS. For this purpose, 77 Holstein-Friesian heifers were randomly assigned to one of 2 experimental groups: control (CON) or phantom (PHAN). Four weeks before calving, the PHAN group was given free access to the phantom, which was similar to the actual milking robot, so that they could explore it and be positively conditioned by feeding concentrate in the phantom. The heifers of the CON group had no contact with the phantom or the AMS before the first milking at the AMS. The milking frequency per animal per day was recorded, and the proportion of animals that had to be fetched for milking was determined, to evaluate how the animals accepted the AMS after calving. To assess the stress level of the animals before and after introduction into the AMS, fecal cortisol concentrations and rumination times of the animals were measured. Additionally, lactation performance characteristics (milk yield, milk flow, electrical conductivity of milk, and milk composition) were recorded for 77 animals. The animals trained on the phantom showed a higher milking frequency (DIM 7: 2.70 ± 0.14 visits/d) than the control animals (DIM 7: 2.41 ± 0.14 visits/d) between the 4th and 10th day of lactation. In addition, between d 1 and d 5, the proportion of animals that had to be fetched for milking was lower in PHAN (DIM 1: 35.18 ± 4.16%) than in CON (DIM 1: 48.03 ± 4.46%). The PHAN heifers had unexpectedly high fecal cortisol levels (1 wk prepartum: 43.50 ± 0.93 ng/g of feces), although not considerably elevated compared with CON (1 wk prepartum: 40.76 ± 1.05 ng/g of feces). Training on the phantom had no appreciable influence on rumination time and lactation performance parameters. The increased number of milking visits and the reduced proportion of animals that had to be fetched into the AMS for milking indicate that training on the phantom prepares the animals well for being milked in the AMS. Therefore, training heifers on the phantom offers the possibility to facilitate the start into early lactation for the animals, providing a valuable contribution to improvement of animal welfare.

Cortisol-Related Signatures of Stress in the Fish Microbiome.

Exposure to environmental stressors can compromise fish health and fitness. Little is known about how stress-induced microbiome disruption may contribute to these adverse health effects, including how cortisol influences fish microbial communities. We exposed juvenile Atlantic salmon to a mild confinement stressor for two weeks. We then measured cortisol in the plasma, skin-mucus, and feces, and characterized the skin and fecal microbiome. Fecal and skin cortisol concentrations increased in fish exposed to confinement stress, and were positively correlated with plasma cortisol. Elevated fecal cortisol was associated with pronounced changes in the diversity and structure of the fecal microbiome. In particular, we identified a marked decline in the lactic acid bacteria Carnobacterium sp. and an increase in the abundance of operational taxonomic units within the classes Clostridia and Gammaproteobacteria. In contrast, cortisol concentrations in skin-mucus were lower than in the feces, and were not related to any detectable changes in the skin microbiome. Our results demonstrate that stressor-induced cortisol production is associated with disruption of the gut microbiome, which may, in turn, contribute to the adverse effects of stress on fish health. They also highlight the value of using non-invasive fecal samples to monitor stress, including simultaneous determination of cortisol and stress-responsive bacteria.

Roe deer stress response to a wind farms: Methodological and practical implications

There is an insufficient number of field studies on the impact of wind farms on ungulates despite the rapid expansion of world wind energy. The results of previous studies suggests a possible impact of power plants, but they are divergent and do not take into account the size of wind farms. We examined the impact of wind farms on the stress levels in roe deer based on seven wind farms of various sizes (of 12–27 turbines) in eastern Poland. Fecal cortisol concentration was assessed with the ELISA method in droppings collected during the winter period. We found that the roe deer exhibited an elevated stress level in the area of larger farms, but such response was not found in the case of smaller wind farms. The roe deer were also characterized by a higher stress level in areas of wolf predation, but this factor was less important. Both the area of the farm and the number of turbines explained the phenomenon of the increased stress in the roe deer. We estimated 824 ha or 18 turbines as a threshold level of the impact of wind farms on the cortisol concentration in the roe deer. We conclude, that turbines should be concentrated in the smallest possible area. In such conditions, roe deer will probably be able to find appropriate refuges. We recommend further studies on wind farms older than 4 years and the distribution of turbines.

Fecal Cortisol and Progesterone Concentrations in Post Partus of Etawah Crossbreed Goat

Progesterone (P4) is a dominant hormone during pregnancy. In the later stage of pregnancy, the stress hormone particularly cortisol (C) may increase for initiating the parturition process as a consequence of fetal stress. This study was a preliminary study to compare the concentration of P4 and C in feces of Etawah Crossbreed Goat during their last stage of pregnancy and post partus. This study used 5 pregnant Etawah Crossbreed Goats (t 20th weeks) of pregnancy. Fecal samples were collected in the 20th week of pregnancy to 2 weeks of postpartum. All fecal samples were then dried using a freeze dryer (Labfreez FD10-MR) for 7 days at -80°C. Afterward, dried feces were pulverized and extracted by using 3ml of methanol 80%. The fecal extract was then analyzed the P4 and C concentrations using the enzyme-linked immunosorbent assay method. Concentrations of P4 and C metabolites in the last stage of pregnancy were 5,506.18 3,396.72 ng/g dry feces and 136,625.83 42,479.22 ng/g feces, respectively. Concentrations of P4 and C metabolites in the 2 weeks postpartum decreased at 669.38 P 643.9 ng/g feces and 110,295 / 14,378, 8 ng/g feces, respectively. It canbe concluded that there was a difference in the fecal progesterone and cortisol concentrations between the last phase of pregnancy and the postpartum phase.

Raging elephants: effects of human disturbance on physiological stress and reproductive potential in wild Asian elephants.

Human disturbance has become a widespread threat to wildlife viability. The Asian elephant (Elephas maximus), an endangered and disturbance-prone species, is under severe threat from habitat loss and fragmentation, human-elephant conflict and poaching. Establishing connections between human disturbance, stress responses and reproduction is crucial for assessing the long-term survivability of a species and will provide critical information for conservation management. The current study investigated the effects of human disturbance on population-level stress responses and stress-related effects on reproductive potential of wild Asian elephants in Xishuangbanna Dai Autonomous Prefecture, China. We used a radioimmunoassay to measure the concentration of fecal cortisol and estradiol in 257 samples collected from five local populations at 15 sites over 4years. Human disturbance in Xishuangbanna was quantified based on the Ecological-Niche Factor Analysis model. We found that fecal cortisol concentrations were strongly positively correlated with the degree of human disturbance and increased markedly with the expansion of tea plantations. Percentage of non-stressed individuals in a population was higher depending on the extend of undisturbed area in their home ranges. Fecal estradiol concentrations decreased significantly with increasing stress levels. Our results suggest that human disturbance poses environmental challenges to wild Asian elephant populations, and chronic exposure to human disturbance could lead to population decline. The study demonstrates the efficacy of non-invasive endocrine monitoring for further informing management decisions and developing conservation strategies.

Optimal foraging and physiological responses to the risk of predation: how fecal cortisol concentrations from trapped Allenby’s gerbil (Gerbillus andersoni allenbyi) relate to foraging under the risk of predation

We studied the influence of manipulating predation risk on Allenby’s gerbil ( Gerbillus andersoni allenbyi) held in a large, outdoor enclosure. We measured giving up densities (GUDs), apprehension, time allocation to foraging, harvest strategy (grab and go (GAG) vs. eat at tray (EAT)), and fecal cortisol concentration. First we established the time necessary for cortisol and corticosterone concentrations to change significantly from baseline after a stressful experience. To do this we collected feces from gerbils 2, 4, 6, or 8 hours after being handled (treatment) or not (control). After 8 h, fecal cortisol, but not corticosterone, concentration was significantly higher in treatment animals. We used the results from the hormone time course experiment to design the predation experiment. We used a dog, trained to harass gerbils, to increase predation risk for the gerbils. We predicted that fecal cortisol concentrations would increase directly in the face of predation risk, or indirectly, due to reduced foraging time because of perceived predation risk that, in turn, leads to increased hunger levels. As predicted, in the presence of a predator, GUDs were higher, time allocation lower, and GAG foraging was used more in treatment animals than in controls, but we found no change in apprehension. There was no difference in cortisol concentration between predator present and no-predator treatments. However, individuals that tended to have higher average fecal cortisol concentrations also tended, on average, to spend more time foraging. This indicates a relationship between stress hormones and optimal foraging. This relationship is potentially causal. While nightly changes in behavior may not be related to stress hormones, over course time scales, stress hormones may be driving gerbils to forage more.

Assessing the utility of urinary and fecal cortisol as an indicator of stress ingolden snub-nosed monkeys (Rhinopithecus roxellana).

Cortisol concentration (CC) is often used as a stress indicator in animals, as high CC is associated with elevated stress levels. During field research, non-invasive methods of measuring CC, such as collection of urine and feces, are superior to using blood samples when monitoring free-ranging animals’ stress levels. However, due to different metabolic pathways, whether CC can be detected in urine and feces to reliably assess stress varies across species. Therefore, it is important to ascertain whether urine and fecal samples are a reliable source for determining CCs and to determine a suitable sampling regime. In this study, we subjected three captive adult golden snub-nosed monkeys (Rhinopithecus roxellana) to a high-stress situation (capture and injection). Urine and feces were collected for four days before and for four days after the manipulations for laboratory analysis. Immunoreactive CC was detected with a commercial enzyme immunoassay (EIA) kit and showed distinct rises. Peak CC values in urine were detected within 5 h, while peak fecal CC ranged between 5 and 24 hours post-interference. These results provide evidence that CC in urine and feces can be used to assess stress levels in the golden snub-nosed monkey. The optimal time frame to collect urinary and fecal samples for CC analysis is within one day of a potential stressful event.

Fecal cortisol radioimmunoassay to monitor adrenal gland activity in the bottlenose dolphin (Tursiops truncatus) under human care

AbstractFecal glucocorticoid measurement is an important noninvasive tool to monitor animal health. A radioimmunoassay (RIA) method was developed to measure fecal cortisol in bottlenose dolphins under human care. The method was used to measure baseline hormone levels and evaluate the adrenal response to environmental challenges in a small number of individual dolphins. The method was validated by precision and accuracy tests and by comparison with liquid chromatography‐mass spectrometry (LC‐MS). The parallelism test suggested few matrix interferences. The assay showed a good degree of precision within assay (CV = 5.4%) and between assays (CV = 4.1%). The RIA significantly correlated with the LC‐MS method (r = 0.838, P < 0.01). The recovery test and the comparison between RIA and LC‐MS suggested that the RIA slightly underestimates fecal cortisol concentrations, although the degree of accuracy was good. This study established that bottlenose dolphins excrete appreciable amounts of fecal cortisol (healthy subjects: 0.2–9.5 ng/g). Therefore, chronic HPA axis activation may be monitored in fecal samples by immunoassays after validating a suitable extraction protocol. The RIA could discriminate conditions of stimulation (pregnancy, parturition, isolation, transportation) and inhibition (diazepam administration) of the HPA axis and may, therefore, be useful for monitoring dolphin well‐being.

Influencia del cautiverio en el comportamiento y en los niveles de cortisol del mono titi gris (Saguinus leucopus)

La especie Saguinus leucopus está catalogada como especie en peligro de extinción debido a su escasa distribución y a la destrucción de su hábitat. Su situación de peligro sirvió de justificación para determinar la influencia del cautiverio en los patrones comportamentales y en los niveles de cortisol. Es pertinente el análisis de cortisol y el comportamental, ya que permiten establecer las conductas propias de la especie en situación de estrés y posibilitan la implementación de estrategias favoreciendo el bienestar de los individuos, su reproducción en cautiverio y reintroducción a la vida natural. Metodológicamente se seleccionaron dos encierros el zoológico de Santacruz (con siete individuos en un núcleo familiar) y el zoológico de Piscilago (con dos individuos reunidos con anterioridad); se realizaron etogramas durante 19 días, contabilizando las frecuencias y secuencias comportamentales. Simultáneamente se estimaron las concentraciones de cortisol fecal por medio del Ensayo por Inmunoabsorción Ligado a Enzimas (ELISA), con el fin de hallar alguna posible asociación con el comportamiento en cuanto a la presencia de conductas repetitivas y el aumento del cortisol como posible indicador de estrés. Nuestros resultados muestran que las conductas estereotipadas fueron exploración y/o reconocimiento y se hallaron los estresores en el comportamiento de locomoción. Finalmente, los valores de cortisol fueron mayores en Piscilago (215,5 ng/g) que en Santacruz (145 ng/g), concluyéndose que las estereotipias están asociadas a los niveles de cortisol, como se muestra en la unidad comportamental Lleva (patrón de locomoción), que mostró una dependencia frente a la hormona y unos altos niveles de ésta.

A two-stage method to approach weaning stress in horses using a physical barrier to prevent nursing

Many domestic horses are weaned through abrupt physical separation of the foal and mare. Deviations from normal behaviours testify that this abrupt method has an adverse effect on the well-being of both mares and foals. To reduce negative changes in behaviour and physiology induced by weaning, a two-stage approach was tested: the first stage achieved nutritional separation by means of an udder cover to prevent nursing, and the second stage of physical separation occurred four days later. Behaviours (standing, walking, running, eating, drinking, nursing, aggressing, vocalizing) of mares and foals weaned either by abrupt physical separation (control; n=8), or two-stage (TS; n=7) were recorded in real time for 8h/d, and triaxial accelerometers recorded lying time of mares and foals 24h/d. Heart rates and global position were recorded on foals during live observations, and fecal samples from mares and foals were collected for cortisol analysis. Behavioural and physiological data were analyzed using a mixed model for repeated measures. No treatment effect was observed in foals, however physical separation induced an increase in vocalizations (0.3±0.13 versus 56.9±39.3 vocalizations/foal/d±SEM; p<0.0001), observations of running (0.31±0.08 versus 3.2±0.25 observations/foal/d±SEM; p<0.0001), observations of aggression (2.4±0.27 versus 7.6±0.52 observations/foal/d±SEM; p<0.0001), and distance travelled (1275±150 versus 4679±497m/d±SEM; p<0.0001). Prior to treatment, foals lay 34.91±12.4min/bout in 8±3.4 bouts/d. During nutritional separation, control foals lay down more than TS foals (p<0.0031). There was no treatment effect on lying duration (p>0.75) or number of lying bouts (p>0.17) upon physical separation, but lying time was significantly less in all foals (p<0.01). Prior to treatment, mares lay an average of 14.16±11.21min/bout in 2±1.7 bouts/d. Mares displayed no difference in lying duration or number of lying bouts between treatments (p>0.99) or upon physical separation (p>0.37). Two-stage mares vocalized less frequently than control mares upon physical separation (7.6±0.6 versus 12.6±0.9 vocalizations/mare/d±SEM; p<0.0001). Fecal cortisol concentrations increased in both mares and foals upon physical separation (146±13.5 versus 193±21.9ng/ml±SEM; p<0.0032). Although some minor reductions in stress response were achieved by use of a two-stage approach to weaning, these results more clearly show the effect on mares and foals to physical separation. Large variation in both behavioural and physiological responses to stress may indicate a role of personality and attachment theory in effective approaches to weaning.

TESTOSTERONE AND CORTISOL LEVEL IN FECAL JAVA DEER (Cervus russa timorensis Mul. &amp; Schl 1844) MALE IN CAPTIVITY BUNDER GUNUNG KIDUL YOGYAKARTA

&lt;p&gt;Common reasons for monitoring hormone concentrations in domestic or wild animals, include breeding management, pregnancy diagnosis, stress assessment, and to diagnosed of endocrine illness. The research goal was to develop non invasive analytical techniques of hormone in Java deer (Cervus russa timorensis Mul. &amp;amp; Schl 1844). The non-invasive methods involve the collection of feces, urine, or even saliva in order to measure the metabolites hormones of interest. Samples obtained from either method are most often analyzed with radioimmunoassay (RIA) or enzyme immunoassay (EIA) technology. Both collection and assay methods have advantages and disadvantages. Choice of method depends on the tractability of the species or individual animal in question and the type of question that the data is expected to answer. We are researching basic questions regarding reproductive endocrinology of male Cervus russa timorensis Mul. &amp;amp; Schl 1844. Understanding C timorensis reproductive physiology will help us better address reproductive challenges in both captive and free-ranging populations. Fecal testosterone and cortisol concentrations were measured in captive male C timorensis. As much as five adult male deer were separated into separate cages. Fecal samples were collected in the morning and evening for 30 days. Sample processed through lyophilization stage, pulverization, solvent extraction with methanol and centrifugation to obtain the supernatant for EIA analysis. EIA test results indicated that content of both testosterone and cortisol could be detected, although the levels were still low. This data show that there is a metabolite in-activism in the fecal. With these results through fecal hormone measurement method (non-invasive) it is possible to be developed for observation and research on the reproductive status of the deer.&lt;/p&gt;&lt;p&gt;&lt;br /&gt;&lt;strong&gt;Keywords&lt;/strong&gt;: Non-invasive, Testosterone, Cortisol, Java deer.&lt;/p&gt;

Fecal Concentrations of Cortisol, Testosterone, and Progesterone in Cotton-Top Tamarins Housed in Different Zoological Parks: Relationships Among Physiological Data, Environmental Conditions, and Behavioral Patterns

The aim of this investigation was to study the welfare of 3 captive groups of cotton-top tamarins housed in different zoological parks. Ethological observations were conducted during 1 year. In addition, fecal samples were collected and the concentrations of glucocorticoids, androgens, and progestogens were measured. Within each group, no significant differences in fecal cortisol concentrations were found between subjects. The fecal concentrations of testosterone and progesterone significantly differed depending on the sexes and ages of the tamarins. A significant association was found among hormone concentrations, exhibit dimensions, and group composition. A highly significant correlation was found between all hormones considered and the space available for each subject. Significant differences in behavioral patterns were observed among groups, including social–individual, affiliative–aggressive, and anogenital–suprapubic scent marking. Correlations between hormone measurements and behaviors were detected. In conclusion, this study confirmed the associations between some behaviors exhibited by these nonhuman primates and both cortisol and testosterone; these data also highlight the role played by progesterone in these behaviors.

Standardization of a non-invasive methodology to measure cortisol in hair of beef cattle

A study was conducted to (a) determine if beef cattle hair contains cortisol at measurable concentrations, and (b) identify the effect of hair location and collection method on hair cortisol concentrations. Hair samples (0.5g) from the head, neck, shoulder, hip, and switch were collected from twelve Angus cross bulls (313.1±14.7kg BW) using two sampling methods: plucking, to ensure collection of the hair follicles; and clipping, using an electric razor to ensure collection of the hair as close as possible to the skin. After two washings with isopropanol, hair samples were ground with a ball mill for 5min at 22Hz, sonicated with methanol for 30min, and incubated on a shaker for 18h, at 50°C and 100rpm. The supernatant was pipetted off and evaporated in a block heater, at 45°C under a stream of nitrogen. Samples were reconstituted with phosphate buffered saline before quantification of cortisol with a competitive immunoassay. The described method was successful in detecting cortisol in all the hair samples, with concentrations ranging from 0.30 to 5.31pg/mg. The intra-assay coefficient of variation (CV) ranged from 3.6% to 6.0%, while the inter-assay CV ranged from 5.4% to 11.2%. The cortisol concentration was greater (P<0.05) in the hair from the tail (1.99±0.189pg/mg) compared with the head and the shoulder (1.14 and 0.82±0.189pg/mg, respectively), and in the hair from the neck and the hip (1.50 and 1.59±0.189pg/mg, respectively) compared with the shoulder (0.82±0.189pg/mg). Cortisol concentration was greater (P<0.01) in hair samples collected by clipping (2.35±0.176pg/mg) than by plucking (1.75±0.176pg/mg). There was a day×location interaction (P=0.01), where the hair from the head, neck and shoulder had a lower cortisol concentration at d 28 than at d 1 of the experiment. Data show a significant positive association between cortisol concentration in saliva samples and its level in hair from the hip (r=0.52) and the tail (r=0.63). There was also a trend for a positive association between fecal glucocorticoid metabolites and cortisol concentration in the hair from the neck and the tail (r=0.46 and 0.47, respectively). Results indicate that hair can be used as matrix to measure cortisol levels in beef cattle. Clipping hair from the tail seems to be the most suitable way for measuring cortisol concentration in hair.