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Keel bone damage (KBD) in laying hens is an important welfare problem in both conventional and organic egg production systems. We aimed to identify possible risk factors for KBD in organic hens by analysing cross-sectional data of 107 flocks assessed in eight European countries. Due to partly missing data, the final multiple regression model was based on data from 50 flocks. Keel bone damage included fractures and/or deviations, and was recorded, alongside with other animal based measures, by palpation and visual inspection of at least 50 randomly collected hens per flock between 52 and 73 weeks of age. Management and housing data were obtained by interviews, inspection and by feed analysis. Keel bone damage flock prevalences ranged from 3% to 88%. Compiled on the basis of literature and practical experience, 26 potential associative factors of KBD went into an univariable selection by Spearman correlation analysis or Mann–Whitney U test (with P

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.

Most observers in behaviour studies are aware of relevant information about the animals being observed. We investigated whether observer expectations influence subjective scoring methods during a class practicum. Veterinary students were trained in recording negative and positive interactions between pigs, in scoring the degree of panting in cattle and in applying qualitative behaviour assessment (QBA) using a fixed set of terms for assessing hens' behaviour. The students applied these methods in three trials in which they were shown duplicated video recordings of the same animals: the original and a slightly modified version (to prevent recognition at second viewing). When scoring the duplicated recordings they were told either correct or false information about the conditions in which the animals had been filmed. The false information reflected plausible study scenarios in ethology and was used to create expectations about the outcome. As in reality the students scored the identical behaviour twice, the difference in the scores for the original and modified recordings reflects expectation bias due to providing different contextual information. In all trials there was evidence of expectation bias: students scored the ratio of positive to negative interactions higher when told that the observed pigs had been selected for high social breeding value, they scored cattle panting higher when told that the ambient temperature was 5 °C higher than in reality, and in the QBA they indicated more positive and fewer negative emotions when told that the hens were from an organic instead of a conventional farm. The magnitude of the bias in the QBA trial was related to the opinion of the students about hen welfare in organic versus conventional farms. Although veterinary students may not be representative of practising ethologists, these findings do indicate that observer bias could influence subjective scores of animal behaviour and welfare.

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.

Aviary systems for laying hens offer space and opportunities to perform natural behaviors. However, hen welfare can be impaired due to increased risk for keel bone and foot pad disorders in those systems. This cross-sectional study (N = 47 flocks) aimed to assess prevalences of keel bone and foot pad disorders in laying hens housed in aviaries in Belgium to identify risk factors for these disorders and their relation to egg production. Information on housing characteristics and egg production were obtained through questionnaire-based interviews, farm records, and measurements in the henhouse. Keel bone (wounds, hematomas, fractures, deviations) and foot pad disorders (dermatitis, hyperkeratosis) were assessed in 50 randomly selected 60-week-old laying hens per flock. A linear model with stepwise selection procedure was used to investigate associations between risk factors, production parameters, and the keel bone and foot pad disorders. The flock mean prevalences were: hematomas 41.2%, wounds 17.6%, fractures 82.5%, deviations 58.9%, hyperkeratosis 42.0%, dermatitis 27.6%, and bumble foot 1.2%. Identified risk factors for keel bone disorders were aviary type (row vs. portal), tier flooring material (wire mesh vs. plastic slats), corridor width, nest box perch, and hybrid. Identified risk factors for foot pad disorders were aviary type (row vs. portal), free-range, and hybrid. Percentage of second-quality eggs was negatively associated with keel bone deviations (P = 0.029) at the flock level. Keel bone and foot pad disorders were alarmingly high in aviary housing. The identification of various risk factors suggests improvements to aviary systems may lead to better welfare of laying hens.

Helminths are associated with health- and welfare problems in organic laying hens. The present observational cross-sectional study therefore aimed to estimate the prevalence and worm burdens of intestinal helminths in organic flocks of laying hens in 8 European countries, and to identify management factors that might be associated with helminth infections, with emphasis on Ascaridia galli. Data on flock-level management factors (e.g. nutritional factors, litter quality, housing system, opening- and closing hours of popholes, pasture rotation and provision of occupational materials) were collected during a farm visit when the hens were on average 62 weeks old. Worm counts were performed for 892 hens from 55 flocks and the number of ascarid (presumably primarily A. galli) eggs per g faeces (EPG) for 881 hens from 54 flocks. The association between parasitological parameters (prevalence, worm burden and EPG) and the management factors were analysed by multivariate models. Results showed that A. galli was highly prevalent across Europe with an overall mean prevalence of 69.5% and mean worm burden of 10 worms per hen. The overall mean prevalence and worm burden for Heterakis spp. were 29.0% and 16 worms per hen, respectively, with a large variation between countries. On average, the hens excreted 576 ascarid EPG. The mean prevalence of Raillietina spp. was 13.6%. A positive correlation was found between mean A. galli worm burden and ascarid EPG. Of the analysed management factors, only pasture access time had a significant negative association with A. galli worm burden which was in contrast to the general belief that outdoor access may increase the risk of helminth infections in production animals. In conclusion, the complexity of on-farm transmission dynamics is thus a challenge when evaluating the relative importance of management factors in relation to helminth infections.

On 1 January 2012, conventional cages for laying hens were banned in the European Union (EU); all egg farmers must now use alternative hen housing systems. In total, 218 Flemish egg farmers were surveyed in 2013 to 2014 regarding which housing systems they currently use, their degree of satisfaction with the system, and how they experienced the transition from conventional cages to an alternative system. The response rate was 58.3% (127 respondents). Of these, 43 (33.9%) were no longer active as an egg farmer, mainly due to the ban on conventional cages. The respondents who were active as egg farmers both before and after the transition (84, 66.1%) mainly judged the ban as negative for their own finances and for the competitive position of the Belgian egg industry, but were neutral or positive regarding the general consequences for their own business. Most respondents' hens were housed in either aviary systems (47.7%) or in alternative cage systems (38.2%). When choosing a new system, the fit into the farm and consumer demand were the most important factors. Consumer demand was the main reason for choosing a system with free-range access. In general, egg farmers were satisfied with the system they chose, although this differs between systems. When asked to compare the alternative systems to conventional cages, alternatives were judged to be better for hen welfare and consumer demand, but similar or worse for all other aspects, especially labor. Egg farmers previously using conventional cages judged alternative systems more negatively than those who had no prior experience with conventional cages. Farmers who had experience with free-range systems judged these more positively than those without this experience, e.g., for egg consumer demand, profitability, and hen welfare. These results can possibly be extrapolated to other EU countries in which conventional cages were the most common housing system until 2012, and lessons can be drawn from the farmers' experiences when implementing other animal welfare legislation that may require similar far-reaching adaptations for primary production.

Feather pecking and high mortality levels are significant welfare problems in non-cage housing systems for laying hens. The aim of this study was to identify husbandry-related risk factors for feather damage, mortality, and egg laying performance in laying hens housed in the multi-tier non-cage housing systems known as aviaries. Factors tested included type of system flooring, degree of red mite infestation, and access to free-range areas. Information on housing characteristics, management, and performance in Belgian aviaries (N = 47 flocks) were obtained from a questionnaire, farm records, and farm visits. Plumage condition and pecking wounds were scored in 50 randomly selected 60-week-old hens per flock. Associations between plumage condition, wounds, performance, mortality, and possible risk factors were investigated using a linear model with a stepwise model selection procedure. Many flocks exhibited a poor plumage condition and a high prevalence of wounds, with considerable variation between flocks. Better plumage condition was found in wire mesh aviaries (P

In this study, the effect of human erythropoietin Δ (Epo) on smooth muscle cell (SMC)–rich lesions was evaluated. Mice, of which the left carotid artery was ligated, were treated with suberythropoietic as well as erythropoietic doses of Epo and both doses of Epo enhanced SMC-rich lesion formation. No association was observed between hemoglobin levels and lesion size. Moreover, endothelial progenitor cell (EPC) numbers in the peripheral blood increased only in the erythropoietic dosing group, indicating that EPC numbers did not correlate with lesion size. Immunohistochemical analysis revealed that Epo-mediated enhancement of lesion formation correlates with increased signal transducer and activator of transcription 5 (Stat5) phosphorylation in the vessel wall. Experiments performed in cultured vascular cells demonstrated that Epo robustly induced phosphorylation of Stat5 in human umbilical vein endothelial cells (HUVECs), but only very weakly in SMCs. In tumor necrosis factor-α (TNFα)–activated HUVECS, Epo induced expression of platelet-derived growth factor B (PDGF-B), which was at least partially responsible for the induction of Stat5 phosphorylation in SMCs by HUVEC-conditioned medium. In conclusion, in mice Epo accelerates SMC-rich neointima formation, which correlates with increased Stat5 phosphorylation in the vessel wall but is independent of erythrocyte and EPC numbers.

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.

Simple Summary Until 2012, laying hens in the EU were often housed in conventional cages that offered limited space and few opportunities to perform highly motivated behaviors. Conventional cages are now banned in the EU in order to improve animal welfare. In this study, egg farmers were surveyed (winter 2013–2014) to assess whether they perceived any changes in animal welfare since changing housing systems, what role hen welfare played in choosing a new housing system, and which aspects of hen welfare they find most important. The data show that the answers differ depending on which housing system the farmers currently use and whether they had used conventional cages in the past. Abstract As of 2012, the EU has banned the use of conventional cages (CC) for laying hens, causing a shift in housing systems. This study’s aim was to gain insight into farmers’ opinions on hen health and welfare in their current housing systems. A survey was sent to 218 Belgian egg farmers, of which 127 (58.3%) responded, with 84 still active as egg farmer. Hen welfare tended to be less important in choosing the housing system for farmers with cage than with non-cage systems. Respondents currently using cage systems were more satisfied with hen health than respondents with non-cage systems. Reported mortality increased with farm size and was higher in furnished cages than in floor housing. Feather pecking, cannibalism, smothering and mortality were perceived to be higher in current housing systems than in CC, but only by respondents who shifted to non-cage systems from previously having had CC. Health- and production-related parameters were scored to be more important for hen welfare as compared to behavior-related parameters. Those without CC in the past rated factors relating to natural behavior to be more important for welfare than those with CC. This difference in opinion based on farmer backgrounds should be taken into account in future research.