Effects of external ambient temperature at loading, journey duration and flock characteristics on the dead-on-arrival rate in broiler chickens transported to slaughter in Great Britain.

Broiler mortality during transport and lairage, prior to slaughter, has negative welfare and economic implications. Knowledge of the factors affecting the dead-on-arrival (DOA) rate can help identify risk-mitigating strategies. The objectives of this study were to determine the DOA rate in broiler chickens transported to slaughter in Great Britain and associated risk factors. Requested data for all loads of broilers transported to slaughter by 5 large British commercial companies on 57 randomly-selected dates in 2019 were obtained and combined with weather data extracted from the Met Office MIDAS Open database. The DOA rate was described overall and per load using summary descriptive statistics. Mixed-effects Poisson regression was used to evaluate considered flock-, journey- and weather-related risk factors. Results were reported as incidence rate ratios (IRR) and 95% confidence intervals (CI). On the selected dates, 25,476 loads transported 146,219,189 broilers to slaughter. The overall mean DOA rate was 0.08%. The median DOA rate per load was 0.06% (interquartile range 0.03-0.09%; range 0.00-17.39%). Multiple risk factors were identified including loading temperature and catch method. At relative humidity ≤80%, the DOA rate was 16.89 (95% CI 15.25-18.70, P < 0.001) times higher for loads loaded in external ambient temperatures >30.0°C compared to those loaded in temperatures between 10.1°C and 15.0°C. When relative humidity was >80%, there was a 43% increase in DOA rate for loads loaded in temperatures below freezing compared to those loaded in temperatures between 10.1°C and 15.0°C (IRR 1.43, 95% CI 1.35-1.52, P < 0.001). The DOA rate was 32% higher for loads caught mechanically compared to those caught manually (IRR 1.32, 95% CI 1.23-1.42, P < 0.001). The overall DOA rate was lower than that previously reported in Great Britain and for other European countries. Most identified risk factors had a marginal effect, however, loading temperatures >30°C substantially increased DOA rate. Internal thermal environmental conditions were not evaluated. Avoidance of loading during periods of hot weather would improve the welfare of, and reduce economic losses in, broiler chickens.

Associations between behaviour and health outcomes in conventional and slow-growing breeds of broiler chicken.

Broiler chickens are prone to a range of complex health and welfare issues. To support informed selection of welfare traits whilst minimising impact on production efficiency and to address a major gap in understanding, we systematically explored associations between health and behavioural indicators of broiler welfare. One conventional (CNV, n = 350) and two slow-growing broiler breeds (SGH and SGN, respectively n = 400) were reared from hatch in pens of 50 birds. Birds were assessed for health (gait, plumage cover and dirtiness, pododermatitis, hockburn, and leg deviations) at 2.2 kg liveweight according to the Royal Society for the Prevention of Cruelty to Animals Broiler Breed Welfare Assessment Protocol. Behaviour and resource-use of 10% of birds per pen, on days 29 (all breeds) and 43 (SGH and SGN), was (i) scan sampled every 60 min between three to six and between twelve to fifteen hours after photoperiod onset; and (ii) continuously sampled sequentially from focal birds for 3 min each in a random order, during 15 min observation periods at three and twelve hours after photoperiod onset. Binary logistic generalized linear models were used, to assess respective associations between pen prevalence of each health outcome and (i) pen mean percentage scans of behaviour, and (ii) pen mean frequency and duration per 3 min focal observation of behaviour. Better growth rate and feed conversion but poorer health outcomes (mortality, gait, pododermatitis, feather cover) were more prevalent in CNV. Strong associations between behaviour and several heath indicators revealed, (i) increases in side-lying inactive, sitting inactive, and use of the litter relative to other resources, as primary and general indicators of poorer health, and (ii) increases in standing inactive, perch use, walking, Comfort, High Energy and Exploratory behaviour as primary and general indicators of better health. Of these, changes in side-lying, standing inactive, walking, Comfort and High Energy behaviour were particularly sensitive to small differences in health outcomes important for breed acceptance in high-welfare schemes. Crucially these behavioural measures additionally represent motivational and affective aspects of welfare not captured by health measures and allow opportunity for earlier intervention. Thus, to provide a comprehensive assessment of broiler experience, behaviour should be incorporated into broiler welfare assessments.

Light-based monitoring devices to assess range use by laying hens.

Access to an outdoor range has many potential benefits for laying hens but range use can be poor due to factors only partly understood. Techniques to monitor individual range use within commercial flocks are crucial to increase our understanding of these factors. Direct observation of individual range use is difficult and time-consuming, and automatic monitoring currently relies on equipment that is difficult to use in an on-farm setting without itself influencing range use. We evaluated the performance of a novel small, light and readily portable light-based monitoring system by validating its output against direct observations. Six commercial houses (2000 hens/house) and their adjacent ranges were used, three of which were equipped with more structures on the range than the others (to determine whether cover would influence monitoring accuracy). In each house, 14 hens were equipped with light monitoring devices for 5 discrete monitoring cycles of 7 to 8 consecutive days (at 20, 26, 32, 36 and 41 weeks of age). Light levels were determined each minute: if the reading on the hen-mounted device exceeded indoor light levels, the hen was classified as outside. Focal hens were observed directly for 5 min/hen per week. Accuracy (% of samples where monitoring and direct observations were in agreement) was high both for ranges with more and with fewer structures, although slightly better for the latter (92% v. 96% ± 1 SEM, F1,19 = 5.2, P = 0.034). Furthermore, accuracy increased over time (89%, 94%, 95%, 98% ± 1 SEM for observations at 26, 32, 36 and 41 weeks, respectively, F3,19 = 3.2, P = 0.047), probably due to progressively reduced indoor light levels resulting from partial closing of ventilation openings to sustain indoor temperature. Light-based monitoring was sufficiently accurate to indicate a tendency for a greater percentage of monitored time spent outside when more range structures were provided (more: 67%, fewer: 56%, SEM: 4, $\chi_1^2 = 2.9$, P = 0.089). Furthermore, clear and relatively consistent individual differences were detected. Individuals that were caught outside at the start of the experiment ranged more throughout its duration (caught outside: 72%, caught inside 51%, SEM: 4, $\chi_1^2 = 10.0$, P = 0.002), and individual range use was correlated between monitoring cycles (for adjacent monitoring cycles: $r_s^2 = 0.5-0.7$, P < 0.0001). This emphasizes the importance of studying range use on an individual level. In conclusion, our light-based monitoring system can assess individual range use accurately (although accuracy was affected by house characteristics to some extent) and was used to show that both cover availability and individual characteristics affected range use.

A PKA/cdc42 Signaling Axis Restricts Angiogenic Sprouting by Regulating Podosome Rosette Biogenesis and Matrix Remodeling.

Angiogenic sprouting can contribute adaptively, or mal-adaptively, to a myriad of conditions including ischemic heart disease and cancer. While the cellular and molecular systems that regulate tip versus stalk endothelial cell (EC) specification during angiogenesis are known, those systems that regulate their distinct actions remain poorly understood. Pre-clinical and clinical findings support sustained adrenergic signaling in promoting angiogenesis, but links between adrenergic signaling and angiogenesis are lacking; importantly, adrenergic agents alter the activation status of the cAMP signaling system. Here, we show that the cAMP effector, PKA, acts in a cell autonomous fashion to constitutively reduce the in vitro and ex vivo angiogenic sprouting capacity of ECs. At a cellular level, we observed that silencing or inhibiting PKA in human ECs increased their invasive capacity, their generation of podosome rosettes and, consequently, their ability to degrade a collagen matrix. While inhibition of either Src-family kinases or of cdc42 reduced these events in control ECs, only cdc42 inhibition, or silencing, significantly impacted them in PKA(Cα)-silenced ECs. Consistent with these findings, cell-based measurements of cdc42 activity revealed that PKA activation inhibits EC cdc42 activity, at least in part, by promoting its interaction with the inhibitory regulator, guanine nucleotide dissociation inhibitor-α (RhoGDIα).

A LATS biosensor screen identifies VEGFR as a regulator of the Hippo pathway in angiogenesis.

The Hippo pathway is a central regulator of tissue development and homeostasis, and has been reported to have a role during vascular development. Here we develop a bioluminescence-based biosensor that monitors the activity of the Hippo core component LATS kinase. Using this biosensor and a library of small molecule kinase inhibitors, we perform a screen for kinases modulating LATS activity and identify VEGFR as an upstream regulator of the Hippo pathway. We find that VEGFR activation by VEGF triggers PI3K/MAPK signaling, which subsequently inhibits LATS and activates the Hippo effectors YAP and TAZ. We further show that the Hippo pathway is a critical mediator of VEGF-induced angiogenesis and tumor vasculogenic mimicry. Thus, our work offers a biosensor tool for the study of the Hippo pathway and suggests a role for Hippo signaling in regulating blood vessel formation in physiological and pathological settings.

Internal roosting location is associated with differential use of the outdoor range by free-range laying hens.

1. In commercial free-range systems for laying hens, popholes to the outdoor range are often installed on one side of the house only. In multi-tier systems, it is possible that some individuals fail to access the range due to internal barriers to movement. 2. Five commercial multi-tier flocks from different units were studied. For each flock, two different colour markers were used to distinguish 200 birds roosting near the popholes (NP-Roost) and 200 birds roosting far from the popholes (FP-Roost) at night. The following day, counts of marked birds on the range and inside the house were performed. 3. Significantly more NP-Roost birds were observed in all areas of the outdoor range than FP-Roost birds the next day. Distance of FP area from the popholes was very strongly positively correlated with effect size in the adjacent range area. 4. Additionally, in the indoor area far from the popholes (FP) more FP-Roost birds were observed the next day than NP-Roost birds. In the indoor area near to the popholes (NP) more NP-Roost birds were observed the next day than FP-Roost birds. 5. These results suggest that roosting location is associated with differential range use when popholes are only available on one side of the shed as birds that roosted far from the popholes used the range less.

Short communication: Detection of lameness in dairy cows using a grooming device.

Lameness in dairy cattle is a common welfare problem with significant economic implications. All too often, appropriate treatment is delayed or neglected due to insufficient detection of lame cows. Brush usage is considered a low-resilience activity; that is, one that typically decreases when energy resources are limited or when the cost involved in the activity increases, such as during sickness and stress. The aim of this study was to determine the association between brush usage and different degrees of lameness. Locomotion scores of 209 lactating Holstein dairy cows were collected individually once a week for 14 consecutive weeks, using a 5-point visual assessment scoring system (1 = nonlame, 2 = uneven gait, 3 = mild lameness, 4 = lameness, 5 = severe lameness). Daily brush usage was collected automatically from 3 cowsheds of similar size and structure located on a commercial dairy farm. In each of the 3 cowsheds, 2 brushes were installed, one next to the feed bunk, and the other away from the feed bunk (on the opposite side of the cowshed). Linear and generalized linear mixed-effects models were used to evaluate the association between locomotion scores and daily measures of brush usage. We found a significant interaction between locomotion score and brush location (near to/distant from feed bunk) on the daily proportion of cows using the brush at least once and on daily duration of brush usage. Specifically, we showed that lame and severely lame cows did not use brushes that were installed away from the feed bunk but continued to use brushes that were installed next to the feed bunk. Brush usage by cows with uneven gait (locomotion score 2) or with mild lameness (locomotion score 3) did not differ from that of nonlame cows (locomotion score 1). The results of this study suggest that monitoring of daily usage of brushes located away from the feed bunk could be a useful method for detecting lameness and severe lameness in dairy cows. However, the use of this method to detect mild lameness or cases of abnormal gait is, at this stage, less promising.

Short communication: Detection and monitoring of metritis in dairy cows using an automated grooming device.

Metritis, a prevalent disease on dairy farms, is negatively associated with reproduction, milk production, and the welfare of cows. The objective of this study was to evaluate the efficacy of monitoring low-resilience activities (i.e., behaviors that typically decrease when energy resources are limited or when the cost involved in the activity increases; e.g., brush usage) in the early detection of metritis. Data on daily brush usage (i.e., proportion of cows using the brush and the duration of usage) were collected from 28 metritic and 60 control cows 28 d postpartum using an automated monitoring system developed for the purpose of this study. During the first week following partum (before clinical diagnosis), we found no differences in brush usage between sick and control cows. However, 8 to 21 d postpartum (the week of clinical diagnosis and the first week of medical treatment), a lower proportion of metritic cows used the brush compared with control cows (0.49 compared with 0.64, respectively, at brushes installed away from the feed bunk). In addition, the daily duration of brush usage was 50% lower among cows diagnosed with metritis compared with control cows 8 to 28 d postpartum (44 s/d compared with 88 s/d, respectively). The results of this study suggest that on-farm monitoring of low-resilience behaviors, combined with existing systems that monitor core behaviors (e.g., activity and rumination), may serve as an improved method for detecting events that compromise the welfare of animals. The slow recovery of low-resilience behaviors following medical treatment (wk 4) might serve as a particularly useful indicator of progress of recovery from disease.

Invited review: Environmental enrichment of dairy cows and calves in indoor housing.

In recent years, an increasing number of farmers are choosing to keep their cows indoors throughout the year. Indoor housing of cows allows farmers to provide high-yielding individuals with a nutritionally balanced diet fit for their needs, and it has important welfare benefits for both cows and their calves, such as protection from predators, parasites, and exposure to extreme weather conditions. However, it also confronts cows and calves with a wide range of environmental challenges. These include abiotic environmental sources of stress (e.g., exposure to loud and aversive sound) and confinement-specific stressors (e.g., restricted movement and maintenance in abnormal social groups). Cows and calves that live indoors are also faced with the challenge of occupying long periods with a limited range of possible behavioral patterns. Environmental enrichment can improve biological functioning (measured as increased lifetime reproductive success, increased inclusive fitness, or a correlate of these such as improved health), help animals to cope with stressors in their surroundings, reduce frustration, increase the fulfillment of behavioral needs, and promote more positive affective states. Here, we review recent findings on the effect of social, occupational, physical, sensory, and nutritional enrichment on dairy cows and calves, and we assess the appropriateness and practicality of implementing different enrichment practices on commercial dairy farms. Some of the enrichment methods reviewed here may also be applied to those more extensive cattle-raising systems, where similar challenges occur.

Elevated arousal at time of decision-making is not the arbiter of risk avoidance in chickens.

The somatic marker hypothesis proposes that humans recall previously experienced physiological responses to aid decision-making under uncertainty. However, little is known about the mechanisms used by non-human animals to integrate risk perception with predicted gains and losses. We monitored the behaviour and physiology of chickens when the choice between a high-gain (large food quantity), high-risk (1 in 4 probability of receiving an air-puff) option (HGRAP) or a low-gain (small food quantity), no-risk (of an air-puff) (LGNAP) option. We assessed when arousal increased by considering different stages of the decision-making process (baseline, viewing, anticipation, reward periods) and investigated whether autonomic responses influenced choice outcome both immediately and in the subsequent trial. Chickens were faster to choose and their heart-rate significantly increased between the viewing and anticipation (post-decision, pre-outcome) periods when selecting the HGRAP option. This suggests that they responded physiologically to the impending risk. Additionally, arousal was greater following a HGRAP choice that resulted in an air-puff, but this did not deter chickens from subsequently choosing HGRAP. In contrast to human studies, we did not find evidence that somatic markers were activated during the viewing period, suggesting that arousal is not a good measure of avoidance in non-human animals.

Factors affecting ranging behaviour in young and adult laying hens.

1. A study was conducted to investigate the effect of environment on ranging in 33 flocks reared with (16) or without (17) range access. Ranging was observed at 8, 16 and 35 weeks. Information on house layout, weather conditions and range characteristics was used to create models predicting the percentage of the flock out on the range and the percentage of ranging birds observed away from the house. 2. Three flocks had range access at 8 weeks. The percentage of birds ranging averaged 28%, with 22% of these ranging away from the house. For the 13 flocks with range access at 16 weeks, the percentage of pullets on the range was 12%, with 29% of these ranging away from the house. At 35 weeks, all flocks had range access and the average percentage of birds out on the range was 13%, with 42% of these ranging away from the house. 3. The percentage of birds seen using the range was higher with reduced flock size and stocking density, increased pop hole availability (cm/bird) and light intensity inside the house. More birds ranged on cooler days and on farms located in areas with fewer days of rain per year and lower average rainfall. The percentage of birds ranging varied with season and was lowest in May. More birds ranged away from the house when cover and more artificial structures were present on the range. The proportion of ranging birds located away from the house increased with lower outdoor humidity levels, higher air pressure, and on warmer days. Lastly, birds ranged away from the house more as they got older.

Behavioural and physiological expression of arousal during decision-making in laying hens.

Human studies suggest that prior emotional responses are stored within the brain as associations called somatic markers and are recalled to inform rapid decision-making. Consequently, behavioural and physiological indicators of arousal are detectable in humans when making decisions, and influence decision outcomes. Here we provide the first evidence of anticipatory arousal around the time of decision-making in non-human animals. Chickens were subjected to five experimental conditions, which varied in the number (one versus two), type (mealworms or empty bowl) and choice (same or different) of T-maze goals. As indicators of arousal, heart-rate and head movements were measured when goals were visible but not accessible; latency to reach the goal indicated motivation. We found a greater increase in heart-rate from baseline to the goal-viewing period, more head movements and shorter latencies in all conditions including mealworms compared to those with empty bowls. More head movements when two mealworm bowls were available compared to just one, and prior to occasions when hens accessed an empty bowl rather than declining to move, showed that arousal preceded and influenced decision-making. Our results provide an important foundation for investigating arousal during animal decision-making and suggest that the somatic-marker hypothesis might not only apply to humans.

The effect of food location, heat load, and intrusive medical procedures on brushing activity in dairy cows.

Animals allocate time and effort to a range of core (e.g., sleeping, feeding, drinking) and "luxury" (e.g., playing, exploring) activities. A luxury activity is characterized by low resilience and, as such, will be reduced when time or energy resources are limited, including under conditions of stress or discomfort. One seemingly luxurious activity available to cows on an increasing number of dairy farms is rubbing against an automated brush. The current study examined the effect of distance from food, heat load, and an intrusive medical procedure (i.e., artificial insemination and transrectal pregnancy examination) on the resilience of brush usage. The probability of using the brush decreased significantly when food was located distantly from the brush (mean=0.53) compared with days when food was located closer to the brush (mean=0.81). Brush usage also decreased at high temperature and humidity levels, with an average decrease of 0.062 brushing events for an increase of 1 temperature-humidity index unit (95% confidence interval=-0.93-0.030). In addition, a significant reduction of approximately 50% in brushing activity was observed on days of artificial insemination compared with the preceding 3d and the following 3d. These findings show that brush usage is a low resilience activity that reduces under a range of conditions. It may thus have the potential to be used as an indicator of a range of health and welfare problems in cows. Further research should be conducted to assess the sensitivity and specificity of this suggested tool and its possible contribution to the early detection of morbidity.

Surface temperature changes in response to handling in domestic chickens.

Stress-induced hyperthermia (SIH) occurs in numerous species and is characterised by an increase in core body temperature, and a decrease in surface temperature, of between 0.5 and 1.5°C within 10 to 15 min of the onset of "emotional stress". The aim of the current study was to ascertain whether the husbandry-relevant procedure of handling resulted in measurable changes in surface body temperature in chickens, as measured using infrared thermography. Baseline temperatures for 19 domestic hens were compared to temperatures immediately, and up to 20 min following handling (catching and brief restraint by a human). Surface head, eye and comb temperatures were plotted to investigate the pattern of temperature change. In response to handling, comb temperature decreased significantly, showing a rapid 2°C drop. Eye temperature showed an initial decrease then rose to levels significantly higher than handling. Head temperature increased over the 20 min post-handling period, to reach levels significantly higher than baseline. It can be concluded that surface temperature changes assessed using infrared thermography, in particular of the hen's comb, are sensitive to husbandry procedures such as handling and represent a potentially useful method for assessing stress-induced hyperthermia in chickens.

The effect of keel fractures on egg production, feed and water consumption in individual laying hens.

The impact of keel bone fractures on egg production, egg weight and feed and water consumption in individual laying hens. A total of 165 Lohmann brown laying hens were obtained from a commercial farm that consisted of 105 with keel fractures and 60 without keel fractures. 2. After a 4-d period of acclimatisation, hens were individually housed and provided with ad libitum food and water for a 24-h period. The number of eggs laid, egg weight, feed and water consumption during this period were recorded. Keel bone strength was also assessed. 3. Hens free from keel fractures laid more eggs (91.7% vs. 84.9%) of significantly heavier weight (61.9 g vs. 60.2 g), ate less feed (139 g vs. 151 g) and drank less water (212 ml vs. 237 ml) than hens with fractures. 4. There was a significant positive association between keel fracture severity and water consumption, and a significant negative association between keel fracture severity and egg weight and keel bone strength. 5. This small-scale study on individual birds shows that keel bone fractures may have an impact on the economics of egg production.

A bespoke management package can reduce levels of injurious pecking in loose-housed laying hen flocks.

This study investigated the protective effects of an on-farm management package designed to reduce injurious pecking (IP) in loose-housed laying hens. A systematic review of scientific literature generated 46 potentially protective management strategies. Bespoke management packages were designed for treatment flocks (TF) using these management strategies. IP in 53 TFs was compared with IP in 47 control flocks (CF) where the management package was not employed. Scoring of plumage damage (PD) and observations of gentle and severe feather pecking (GFP; SFP), and vent and cannibalistic pecking (VP) were completed, and management strategy use was recorded, at 20, 30 and 40 weeks of age. Differences between treatment and CF were examined using multilevel modelling. Compared with CF, TF employed more management strategies (P<0.001), had lower PD (P=0.003) and SFP (P=0.019). Regardless of treatment or control flock status, the more of the 46 management strategies that were employed the lower was the PD (P=0.004), GFP (P=0.021), SFP (P=0.043), mortality at 40 weeks (P=0.025), and the likelihood of VP (P=0.021). Therefore, the provision of a bespoke management package was protective against the majority of forms of IP in commercial laying hen flocks.

Lame broiler chickens respond to non-steroidal anti-inflammatory drugs with objective changes in gait function: a controlled clinical trial.

Leg health of intensively reared broiler chickens is a significant problem, yet little is known regarding the nature of lameness-associated pain. Kinematic changes in gait have been reported in naturally lame broilers following subcutaneous non-steroidal anti-inflammatory drug (NSAID) administration, compared to a placebo group. In the current study, an extensive range of gait parameters were defined using a commercial motion-capturing system to record three-dimensional temporospatial information from walking broilers pre- and post-treatment. Data analysis, performed using multi-level models, revealed gait modifications in broilers receiving NSAID, but not in those receiving saline. The effect of walking velocity was accounted for retrospectively. An increase in velocity following NSAID treatment (carprofen: P<0.001; meloxicam: P=0.044) indicated improved walking ability. For several measures, the polarity of the treatment effect depended upon walking speed. At slow speeds certain parameters become more like those of non-lame broilers, which may have been indicative of improved stability: stride length and duration (both NSAIDs), transverse back displacement (meloxicam), and vertical leg displacement (carprofen). However, these same parameters also revealed that NSAID treatment caused imbalance at faster speeds, which may have signified an excessive dosage. Although doses employed were not conclusively effective, evidence was provided that factors besides body conformation influenced mobility in the test cohort. The study showed that the model would be useful in future studies to increase our understanding of pain associated with specific lameness types in broiler chickens.

Pop hole use by hens with different keel fracture status monitored throughout the laying period.

The aim of this work was to study the effect of keel fractures on the extent to which free-range hens access the range through pop holes. Over two consecutive laying periods (two production years) a total of 1100 individual birds from one half of a house, divided into four separated flocks, were caught at 25, 35, 45, 55 and 65 weeks, palpated to assess the prevalence and severity of keel fractures and tagged with RFID transponders. Their use of pop holes was subsequently monitored in some cases from week 25 to end of lay at 68 to 70 weeks. At regular intervals (every 10 weeks), the tagged birds were re-caught to assess changes in keel fracture prevalence and severity. The average percentage of birds with fractured keels at 25, 35, 45, 55, 65 and at end of lay (68 to 70 weeks of age) was 5.5, 25.5, 49, 63, 66.5 and 78.5, respectively, across both production years. The effect of keel score on pop hole use was modelled statistically, adjusting for weather conditions and age of the birds. There were significant effects of most of the weather variables recorded, as well as age of the bird, on use of pop holes and also a significant effect of keel score. Higher keel scores resulted in a reduction in pop hole use. A significant statistical interaction between keel score and ambient temperature revealed an accelerated reduction in use as the temperature decreased and keel score increased. It is concluded that the occurrence of keel fractures may affect the birds' ability or willingness to utilise the outdoor range provided by free-range housing systems, thereby reducing the potential welfare advantages of this type of housing.

Development of mechanical and thermal nociceptive threshold testing devices in unrestrained birds (broiler chickens).

Behavioural signs of pain are difficult to quantify and interpret in animals. Nociceptive threshold testing is therefore a useful method for examining the perception and processing of noxious stimuli underlying pain states. Devices were developed to measure response thresholds to quantified, ramped mechanical and thermal nociceptive stimuli applied to the leg or keel of unrestrained birds. Up to 9N mechanical force was delivered via a single round-ended 2mm pin using a pneumatic actuator at 0.4Ns(-1). Heat was applied through a small copper element at 0.8°Cs(-1) to a maximum of 50°C. The repeatability and reliability of threshold measures were validated using 10-12 broiler chickens (aged 49-66 days) per site and modality. Mechanical threshold, or skin and threshold temperature, were recorded over three sessions across a 36h period. Both stimulus types elicited clear, reproducible behavioural responses. Mechanical threshold means and 95% confidence intervals were 3.0 (2.8-3.2)N for keel and 2.0 (1.8-2.1)N for leg sites. Keel thermal tests gave a mean skin temperature of 39.3 (39.1-39.5)°C, and threshold of 46.8 (46.6-47.1)°C. Leg skin temperature was 35.7 (35.6-35.9)°C and threshold 42.5 (42.2-42.8)°C. Threshold measures were consistent within and across sessions and birds showed individual repeatability across tests within sessions. Individual birds' mechanical keel thresholds were also repeatable across sessions. The apparatus gave reliable, reproducible measurements of thresholds to noxious mechanical and thermal stimuli. The range recorded was comparable with previously published nociceptor thresholds in dissected chicken nerve filament fibres, and the method appears suitable for studying nociceptive processes in broiler chickens.

Continuous monitoring of pop hole usage by commercially housed free-range hens throughout the production cycle.

Free-range laying hens are able to move between the indoor house and range through exits termed pop holes. The aim of this study was to examine the proportion of the flock that used the pop holes and to identify patterns of movement throughout the flock cycle. Four flocks of free-range hens each of 1500 birds were studied. Ten per cent of each flock were tagged with RFID (radio-frequency identification) transponders and their pop hole activity studied throughout the production cycle. Within two weeks of tagging at 25, 35, 45, 55 and 65 weeks of age, approximately 80 per cent of the tagged birds were seen in the pop holes and 50 per cent of the tagged birds were seen on 80 per cent of the days available to them after tagging. Within the flock, subpopulations of birds could be identified: those that never ventured to the pop holes (approximately 8 per cent), those that used the pop holes very infrequently (approximately 8 per cent), those that sat in the pop holes (approximately 4 per cent), and those that used the pop holes frequently (approximately 80 per cent). There was an effect of age of the birds, time of day and daily mean temperature on pop hole usage. Additional factors affecting activity on particular days were wind speed, rainfall and hours of sunshine. The findings show that a significant proportion of the flock accesses the pop holes on a regular basis with only a very small proportion preferring to stay in the house.