May Salivary Chromogranin A Act as a Physiological Index of Stress in Transported Donkeys? A Pilot Study

Road transport is known to be a stressful animal husbandry procedure as it induces the activation of two main physiological stress-related pathways: the hypothalamic-pituitary-adrenal cortex axis and the sympathetic-adrenal medulla axis. This preliminary study aimed to investigate whether salivary chromogranin A (CgA) concentration, known as a biomarker of the sympathetic activity system during psychological stress, may represent a novel physiological index of transportation-induced stress in donkeys. Nineteen Romagnolo donkeys, raised in groups on paddocks, were subject to two transportations, following the farm's routine procedures, for a mean duration of 64 min each on two consecutive days. Salivary samples were gently collected 15 min before and 15 min after each transportation. Salivary CgA was measured by a commercially available enzyme-linked immunosorbent assay test. Results showed that CgA salivary levels significantly decreased after both transportations. The physiological mechanisms underlying this result may be related to catestatin activity, a bioactive product of the proteolytic cleavage of CgA, that acts as an inhibitor of catecholamine release. This hypothesis requires further investigation, particularly considering the limited number of subjects involved in this preliminary study. The identification of a reliable and non-invasive stress-marker would represent a useful tool for improving farm animals' welfare in transport conditions

Habituation to transport helps reducing stress-related behavior in donkeys during loading

Adopting proper animal management strategies, including training, might reduce to a substantial extent the adverse effects of transport-related stress in animals. The aim of this study was to evaluate the effect of habituation to transport on stress-related behaviors and physiological indicators during loading and unloading in donkeys. Fourteen donkeys were recruited and divided in two treatment groups: Habituation (H; M = 5, F = 2) and Control (C; M = 5, F = 2). H donkeys were gradually habituated to be transported, traveling together with their mothers and other adult donkeys well-accustomed to transport, while C donkeys had never been transported before. Loading and unloading phases were video recorded and behavior was analyzed. Saliva samples for cortisol concentration determination were collected at rest and after unloading. Latency time to load was significantly shorter for H donkeys than C donkeys (Mann-Whitney; p = 0.004). C donkeys also showed significantly more stress-related behaviors (Mann-Whitney; p = 0.026) and required a higher but not statistically significant number of human interventions to load. Cortisol concentration increased in both groups, but no differences were found between them (Mann-Whitney; p > 0.05). These results suggest that habituation to transport could mitigate stress during loading procedures in donkeys reducing loading time, frequency of stress-related behaviors and diminishing the need of human intervention

The detection of malingering amnesia: an approach involving multiple strategies in a mock crime

The nature of amnesia in the context of crime has been the subject of a prolonged debate. It is not uncommon that after committing a violent crime, the offender either does not have any memory of the event or recalls it with some gaps in its recollection. A number of studies have been conducted in order to differentiate between simulated and genuine amnesia. The recognition of probable malingering requires several inferential methods. For instance, it typically involves the defendant\u2019s medical records, self-reports, the observed behavior, and the results of a comprehensive neuropsychological examination. In addition, a variety of procedures that may detect very specific malingered amnesia in crime have been developed. In this paper, we investigated the efficacy of three techniques, facial thermography, kinematic analysis, and symptom validity testing in detecting malingering of amnesia in crime. Participants were randomly assigned to two different experimental conditions: a group was instructed to simulate amnesia after a mock homicide, and a second group was simply asked to behave honestly after committing the mock homicide. The outcomes show that kinematic analysis and symptom validity testing achieve significant accuracy in detecting feigned amnesia, while thermal imaging does not provide converging evidence. Results are encouraging and may provide a first step towards the application of these procedures in a multimethod approach on crime-specific cases of amnesia

Substrate choice by turkeys differing in 3-point gait-scoring system in a mobility test

Assessment of pain in poultry flocks\u2019 production is one of the biggest welfare concerns. The aim of this study was to evaluate the walking ability and preferences for three types of substrates by turkeys with different degrees of lameness, in order to verify if birds with higher gait score experience pain and how their choices differ from healthy birds. An experimental test was conducted with 16 male-turkeys 116-day-old (B.U.T.6). Nine birds (56.25%) were considered normal (score 0) whereas 7 (43.75%) were evaluated as having mild to severe lameness (scores 1 or 2) according to 3-point gait-score assessment. A familiar (A: wet litter) and unfamiliar (B: plastic slat) substrate were distributed into three horizontal lines (L1, L2, and L3; each one measuring 150cm length x 60cm width) separated by three concrete empty areas of the same measures. L1 was divided into 75cm of A substrate and the same dimensions of B; L2 had 100cm of A and 50cm of B, while L3 had 50cm of A and 100cm of B. The turkeys\u2019 motivation to pass through the lines was triggered by the presence of conspecifics located at the end of the experimental area. Data were analyzed using ANOVA. There were no significant differences between normal and lame turkeys to the total time spent on three lines (215.00\ub161.59s), (233.57\ub174.81s);the total time spent on the empty area (385.22\ub183.70s), (557.71\ub1124.04s) and the total time to cross the testing areas (581.3\ub191.72s), (791.2\ub1154.44s),respectively. Substrate A was chosen by 71.4% of lame birds even in L1 as L2, while 55.6% of normal turkeys preferred substrate B in L1 and 77.8% chose substrate A in L2. All birds walked on substrate B in L3. According to these preliminary results, it seems that lame birds have no difference in substrate choice, or on the time spent in each part of the experiment

Validation of a fear test in sport horses using infrared thermography

The aims of the present study were to assess feasibility and validity of a fear test in adult sport horses and to investigate whether the exposure to a fearful stimulus induces a change in eye temperature. Fifty horses, aged 14 \ub1 6 years, of different breed and gender, entered the study. For each horse, a caretaker was asked to fill in a validated temperament questionnaire. A novel object fear test (NOT), has been selected from literature to examine fearfulness. Temperature of the lacrimal caruncle was measured before the test and after the test on 22 horses, representative of the whole sample. To assess discriminant validity of the NOT, 3 human-animal relationship tests were performed on the same horses. Data were analyzed with descriptive, nonparametric, and multivariate statistic methods. No significant differences were found between females and geldings for any of the measured variables. Horses that were described by caretakers as more prone to panic, vigilant, excitable, skittish, and nervous (P < 0.001) needed significantly longer time to reapproach the novel object (P < 0.01). Eye temperature was significantly higher after the NOT compared to basal (P < 0.01), with subjects who did not reapproach the novel object tending to present larger increases (P < 0.10). Horses showing more fear-related responses to the NOT did not show more negative reactions to humans during the human-animal tests. These results suggest that, to some extent, the NOT predicts horses' behavior in real on-farm situations. Our findings reject the hypothesis that reactivity to humans and general fearfulness belong to the same basic feature of temperament. Importantly, infrared thermography proved to be useful in assessing physiological reactions of fear in horses

Validation of a fear test in sport horses using infrared thermography

The aim of our research was to assess feasibility and validity of a fear test in adult sport horses and to investigate if the exposure to a fearful stimulus induces a change in eye temperature. Fifty horses aged 14\ub16 years of different breed and gender entered the study. A novel object fear test (NOT), modified from G\uf3recka-Bruzda (G\uf3recka-Bruzda et al. 2011, Appl Anim Behav Sci 133:207\u201315), has been selected to examine fearfulness. For each horse, a caretaker was asked to fill in a validated temperament questionnaire. In order to assess discriminant validity of the NOT three human-animal relationship tests were performed on the same horses. Temperature of the lacrimal caruncle was measured pre-test and post-test on 22 horses, representative of the whole sample. Data were analysed with descriptive, non-parametric and multivariate statistic methods. No significant differences were found between female and geldings for any of the measured variables. Horses that were described by caretakers as more prone to panic, vigilant, excitable, skittish and nervous (p < 0.001), needed significantly longer time to approach the novel object (p < 0.01). Eye temperature was significantly higher after the NOT compared to basal (p < 0.01), with subjects who did not re-approach the novel object tending to present larger increases (p < 0.1). Horses showing more fear related responses to the NOT did not show more negative reactions to humans during the human-animal tests. These results suggest that, to some extent, the NOT predicts horses\u2019 behaviour in real on-farm situations. Our findings reject the hypothesis that reactivity to humans and general fearfulness belong to the same basic feature of temperament. Importantly, infrared thermography proved to be useful to assess physiological reactions of fear in horses

Habituation to Transport Helps Reducing Stress-Related Behavior in Donkeys During Loading

Adopting proper animal management strategies, including training, might reduce to a substantial extent the adverse effects of transport-related stress in animals. The aim of this study was to evaluate the effect of habituation to transport on stress-related behaviors and physiological indicators during loading and unloading in donkeys. Fourteen donkeys were recruited and divided in two treatment groups: Habituation (H; M = 5, F = 2) and Control (C; M = 5, F = 2). H donkeys were gradually habituated to be transported, traveling together with their mothers and other adult donkeys well-accustomed to transport, while C donkeys had never been transported before. Loading and unloading phases were video recorded and behavior was analyzed. Saliva samples for cortisol concentration determination were collected at rest and after unloading. Latency time to load was significantly shorter for H donkeys than C donkeys (Mann-Whitney; p = 0.004). C donkeys also showed significantly more stress-related behaviors (Mann-Whitney; p = 0.026) and required a higher but not statistically significant number of human interventions to load. Cortisol concentration increased in both groups, but no differences were found between them (Mann-Whitney; p &gt; 0.05). These results suggest that habituation to transport could mitigate stress during loading procedures in donkeys reducing loading time, frequency of stress-related behaviors and diminishing the need of human intervention