Abstract: Conservation medicine is a discipline in which researchers and conservationists study and respond to the dynamic interplay between animals, humans, and the environment. From a wildlife perspective, animal species are encountering stressors from numerous sources. With the rapidly increasing human population, a corresponding increased demand for food, fuel, and shelter; habitat destruction; and increased competition for natural resources, the health and well-being of wild animal populations is increasingly at risk of disease and endangerment. Scientific data are needed to measure the impact that human encroachment is having on wildlife. Nonbiased biometric data provide a means to measure the amount of stress being imposed on animals from humans, the environment, and other animals. The stress response in animals functions via glucocorticoid metabolism and is regulated by the hypothalamic-pituitary-adrenal axis. Fecal glucocorticoids, in particular, may be an extremely useful biometric test, since sample collection is noninvasive to subjects and, therefore, does not introduce other variables that may alter assay results. For this reason, many researchers and conservationists have begun to use fecal glucocorticoids as a means to measure stress in various animal species. This review article summarizes the literature on many studies in which fecal glucocorticoids and their metabolites have been used to assess stress levels in various mammalian species. Variations between studies are the main focus of this review. Collection methods, storage conditions, shipping procedures, and laboratory techniques utilized by different researchers are discussed.

Abstract: Resistance in the horse to trailer loading is a common source of stress and injury to horses and their handlers. The objective of this study was to determine whether nonaversive training based on the Tellington-Touch Equine Awareness Method (TTEAM; Tellington-Jones &Bruns, 1988) would decrease loading time and reduce stress during loading for horses with a history of reluctance to load. Ten horses described by their owners as “problem loaders” were subjected to pretraining and posttraining assessments of loading. Each assessment involved two 7-min loading attempts during which heart rate and saliva cortisol were measured. The training consisted of six 30-min sessions over a 2-week period during which the horse and owner participated in basic leading exercises with obstacles simulating aspects of trailering. Assessment showed heart rate and saliva cortisol increased significantly during loading as compared to baseline (p <.001 and p <.05, respectively). Reassessment after training showed a decrease in loading time (p <.02), reduced heart rate during loading (p <.002), and reduced saliva cortisol as compared to pretraining assessments. Seven “good loaders” also were subject to loading assessment for physiological comparison. Increases in heart rate during loading were significantly higher in the good loaders (p <.001). Nonaversive training simulating aspects of loading may effectively reduce loading time and stress during loading for horses with a history of resistance to trailer loading.

Abstract: REASONS FOR PERFORMING STUDY: Earlier studies have developed a clinical tool to evaluate objectively the function of the equine back. The ability to differentiate horses with back pain from asymptomatic, fully functioning horses using kinematic measures from this tool has not been evaluated. OBJECTIVES: To compare the kinematics of the back at walk and trot in riding horses with back dysfunction to the same parameters in asymptomatic sport horses. METHODS: The kinematics of the back in 12 horses with impaired performance and back pain were studied at walk and trot on a treadmill. Data were captured for 10 sees at 240 Hz. Range of movement (ROM) and intravertebral pattern symmetry of movement for flexion and extension (FE), lateral bending (LB) and axial rotation (AR) were derived from angular motion pattern data and the results compared to an earlier established database on asymptomatic riding horses. RESULTS: At walk, horses with back dysfunction had a ROM smaller for dorsoventral FE in the caudal thoracic region (T13 = 7.50 degrees, T17 = 7.71 degrees; P<0.05), greater for LB at T13 (8.13 degrees; P<0.001) and smaller for AR of the pelvis (10.97 degrees; P<0.05) compared to asymptomatic horses (FE-T13 = 8.28 degrees, FE-T17 = 8.49 degrees, LB-T13 = 6.34 degrees, AR-pelvis = 12.77 degrees). At trot, dysfunctional horses had a smaller (P<0.05) ROM for FE at the thoracic lumbar junction (T17 = 2.46 degrees, L1 = 2.60 degrees) compared to asymptomatic horses (FE-T17 = 3.07 degrees, FE-L1 = 3.12 degrees). CONCLUSIONS: The objective measurement technique can detect differences between back kinematics in riding horses with signs of back dysfunction and asymptomatic horses. The clinical manifestation of back pain results in diminished flexion/extension movement at or near the thoracic lumbar junction. However, before applying the method more extensively in practice it is necessary to evaluate it further, including measurements of patients whose diagnoses can be confirmed and long-term follow-ups of back patients after treatment. POTENTIAL RELEVANCE: Since the objective measurement technique can detect small movement differences in back kinematics, it should help to clinically describe and, importantly, objectively detect horses with back pain and dysfunction.

Abstract: Ground reaction force (GRF) measurements are often normalised to body mass to facilitate inter-individual comparisons. The objective of this study was to explore the effect of a rider on the GRFs and fetlock joint kinematics of trotting horses. The subjects were 5 dressage-trained horses and 3 experienced dressage riders. Ground reaction force measurements and sagittal view videotapes were recorded as the horses trotted at the same velocity in hand (3.49 +/- 0.52 m/s) and with a rider (3.49 +/- 0.46 m/s). Data were time-normalised to stance duration. Ground reaction force measurements were expressed in absolute terms and normalised to the system mass (horse or horse plus rider). All the horses showed changes in the same direction when comparing the ridden condition with the in-hand condition. There was an increase in the absolute peak vertical GRFs of the fore- and hindlimbs with a rider. However, the mass-normalised peak vertical GRFs were lower for the ridden condition, with the peak occurring later in the forelimbs and earlier in the hindlimbs compared with the inhand condition. Maximal fetlock angle and its time of occurrence were similar for the 2 conditions, but the fore fetlock joint was more extended during the later part of the stance phase in ridden horses. The presence of a rider appeared to affect the GRFs and fetlock joint kinematics differently in the fore- and hindlimbs, and the ridden horse did not seem to be equivalent to a proportionately larger horse. This should be considered when normalising for body mass in studies comparing horses in hand and ridden horses.

Abstract: Non-invasive techniques to monitor stress hormones in small animals like mice offer several advantages and are highly demanded in laboratory as well as in field research. Since knowledge about the species-specific metabolism and excretion of glucocorticoids is essential to develop such a technique, we conducted radiometabolism studies in mice (Mus musculus f. domesticus, strain C57BL/6J). Each mouse was injected intraperitoneally with 740 kBq of 3H-labelled corticosterone and all voided urine and fecal samples were collected for five days. In a first experiment 16 animals (eight of each sex) received the injection at 9 a.m., while eight mice (four of each sex) were injected at 9 p.m. in a second experiment. In both experiments radioactive metabolites were recovered predominantly in the feces, although males excreted significantly higher proportions via the feces (about 73%) than females (about 53%). Peak radioactivity in the urine was detected within about 2h after injection, while in the feces peak concentrations were observed later (depending on the time of injection: about 10h postinjection in experiment 1 and about 4h postinjection in experiment 2, thus proving an effect of the time of day). The number and relative abundance of fecal [3H]corticosterone metabolites was determined by high performance liquid chromatography (HPLC). The HPLC separations revealed that corticosterone was extensively metabolized mainly to more polar substances. Regarding the types of metabolites formed, significant differences were found between males and females, but not between the experiments. Additionally, the immunoreactivity of these metabolites was assessed by screening the HPLC fractions with four enzyme immunoassays (EIA). However, only a newly established EIA for 5alpha-pregnane-3beta,11beta,21-triol-20-one (measuring corticosterone metabolites with a 5alpha-3beta,11beta-diol structure) detected several peaks of radioactive metabolites with high intensity in both sexes, while the other EIAs showed only minor immunoreactivity. Thus, our study for the first time provides substantial information about metabolism and excretion of corticosterone in urine and feces of mice and is the first demonstrating a significant impact of the animals' sex and the time of day. Based on these data it should be possible to monitor adrenocortical activity non-invasively in this species by measuring fecal corticosterone metabolites with the newly developed EIA. Since mice are extensively used in research world-wide, this could open new perspectives in various fields from ecology to behavioral endocrinology.