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There is increasing evidence that the field of applied ethology is prone to expectation biases invalidating research outcomes. Nevertheless, outcome assessors are rarely blinded. We surveyed delegates of the International Society for Applied Ethology (ISAE) 2014 congress shortly before (n = 39 respondents) and after (n = 51 respondents) a combined congress plenary and workshop on expectation bias in applied ethology. The aims were to evaluate the effect of the plenary and workshop on the opinion of applied ethologists in order to better comprehend why blinding outcome assessors seems so rarely practiced as a debiasing technique in this field of research. The results suggest that a moderate awareness about expectancy effects among ethologists and the logistic constraints of blinded observations rather than a perceived low susceptibility of the research field is the larger part of the explanation. Although awareness about expectancy effects and debiasing techniques was higher immediately after than before the congress plenary and workshop, a more sustained and concerted effort is needed throughout all stages of the research process to avoid expectation bias invalidating research finding and to improve the scientific credibility of the field of applied ethology.

Non-cage systems provide laying hens with considerable space allowance, perches and access to litter, thereby offering opportunities for natural species-specific behaviors. Conversely, these typical characteristics of non-cage systems also increase the risk of keel bone and foot pad disorders. The aim of this study was twofold: 1) to investigate if providing ramps between perches (housing factor) reduces keel bone and foot pad disorders and 2) to test for genetic predisposition by comparing 2 different layer hybrids. In a 2 × 2 design, 16 pens were equipped either with or without ramps between perches and nest boxes (8 pens/treatment), and housed with either 25 ISA Brown or Dekalb White birds per pen (in total 200 birds/hybrid). Keel bone injuries and foot health were repeatedly measured via palpation and visual assessment between 17 and 52 wk of age and daily egg production was recorded. The relationships between the dependent response variables (keel bone and footpad disorders, egg production) and independent factors (age, ramps, hybrid) were analyzed using generalized linear mixed models and corrected for repeated measures. Ramps reduced keel bone fractures (F1,950 = 45.80, P

Keel bone damage (KBD) is a critical issue facing the laying hen industry today as a result of the likely pain leading to compromised welfare and the potential for reduced productivity. Recent reports suggest that damage, while highly variable and likely dependent on a host of factors, extends to all systems (including battery cages, furnished cages, and non-cage systems), genetic lines, and management styles. Despite the extent of the problem, the research community remains uncertain as to the causes and influencing factors of KBD. Although progress has been made investigating these factors, the overall effort is hindered by several issues related to the assessment of KBD, including quality and variation in the methods used between research groups. These issues prevent effective comparison of studies, as well as difficulties in identifying the presence of damage leading to poor accuracy and reliability. The current manuscript seeks to resolve these issues by offering precise definitions for types of KBD, reviewing methods for assessment, and providing recommendations that can improve the accuracy and reliability of those assessments.

Feather pecking and cannibalism in chickens can lead to injury or to death of the vic- tims, and is thus a serious welfare and economic issue in modern poultry farming. A sib selection scheme has been initiated to genetically select a low mortality line (LML), which shows decreased mortality due to cannibalism compared to a control line (CL). To determine whether undesired behavioural effects such as impaired cognition, increased fearfulness, or increased anxiety might have inadvertently been co-selected, we tested hens from the fourth generation of selection (n = 9 per selection line) in several behavioural tasks. When tested in a 122 cm × 122 cm open field at age 7 days, the lines showed no differences in locomotion or vocalization. In a T-maze test in which the chickens could navigate to find conspecifics, testing sociality and fearfulness, 12 to 16-day-old CL chickens showed a lack of exploratory behaviour; they did not leave the start box. In contrast, most LML chickens negotiated the maze, and approximately half of them found and stayed close to their con- specifics. This difference points toward higher levels of fearfulness in the CL than LML. In a voluntary approach test assessing fearfulness for humans, conducted when the chicks were 26 days old, the LML approached a familiar human faster, thus displaying lower levels of fearfulness. The same birds were tested in a holeboard test at an age of 25-65 days, the first time this test of spatial memory has been used in an avian species. Our results demonstrated high levels of working memory performance and low levels of reference memory perfor- mance in both lines, with no differences between the lines. Overall, the present results indicate that unwanted behavioural effects were not co-selected with selection on low mortality, and support the feasibility of the use of the LML in farming practice. © 2011 Elsevier B.V. All rights reserved.

Exposure of rats to unpredictable loud sound pulses increases activity of the rate-limiting enzyme for serotonin synthesis, tryptophan hydroxylase (TPH), in the median raphe nucleus (MnR) and a mesolimbocortical serotonergic system. Corticotropin-releasing factor (CRF)-induced activation of a subset of serotonergic neurons in the caudal dorsal raphe nucleus (DR) may underlie stress-related increases in TPH activity in the MnR and a mesolimbocortical serotonergic system. An in vivo acoustic stimulation paradigm and an in vitro brain slice preparation were designed to test the hypothesis that stress-related stimuli and CRF receptor activation have convergent actions on TPH activity in the caudal DR (DRC). We measured 5-hydroxytryptophan (5-HTP) accumulation as an index of TPH activity following inhibition of aromatic amino acid decarboxylase (using NSD-1015). To examine effects of acoustic stimulation on TPH activity, male Wistar rats, pretreated with NSD-1015, were exposed to a 30 min sham, predictable or unpredictable acoustic stimulation paradigm; brains were frozen and microdissected for analyses of tissue 5-HTP concentrations in subregions of the DR. To examine the effect of CRF receptor activation on TPH activity, freshly prepared brain slices were exposed to CRF (0-2000 nM) for 10 min in the presence of NSD-1015, then frozen and microdissected for analysis of tissue 5-HTP concentrations. Increases in TPH activity in the DRC, but not other subregions, were observed in both paradigms. These findings are consistent with the hypothesis that stress-related increases in TPH activity are mediated via effects of CRF or CRF-related neuropeptides on a mesolimbocortical serotonergic system originating in the DRC.

Depressed suicide patients have elevated expression of neuronal tryptophan hydroxylase 2 (TPH2) mRNA and protein in midbrain serotonergic neurons, as well as increases in brain serotonin turnover. The mechanisms underlying these changes are uncertain, but increased TPH2 expression and serotonin turnover could result from genetic influences, adverse early life experiences, or acute stressful life events, all of which can alter serotonergic neurotransmission and have been implicated in determining vulnerability to major depression. Emerging evidence suggests that there are several different stress-related subsets of serotonergic neurons, each with a unique role in the integrated stress response. Here we review our current understanding of how genetic and environmental factors may influence TPH2 mRNA expression and serotonergic neurotransmission, focusing in particular on the dorsomedial part of the dorsal raphe nucleus. This subdivision of the dorsal raphe nucleus is selectively innervated by key forebrain structures implicated in regulation of anxiety states, it gives rise to projections to a distributed neural system mediating anxiety states, and serotonergic neurons within this subdivision are selectively activated by a number of stress- and anxiety-related stimuli. A better understanding of the anatomical and functional properties of specific stress- or anxiety-related serotonergic systems should aid our understanding of the neural mechanisms underlying the etiology of anxiety and affective disorders.