Dorothy E.F. McKeegan

Remote monitoring of electroencephalogram, electrocardiogram, and behavior during controlled atmosphere stunning in broilers: Implications for welfare

This study examined the welfare implications of euthanizing broilers with 3 gas mixtures relevant to the commercial application of controlled atmosphere stunning (CAS). Birds were implanted/equipped with electrodes to measure brain activity (electroencephalogram, EEG) and heart rate. These signals were recorded using a purpose-built telemetry-logging system, small enough to be worn by each bird in a spandex backpack. The birds were euthanized in a scaled-down CAS apparatus consisting of a conveyor belt passing through 2 compartments. Three gas environments were applied (8 birds per treatment): 1) anoxia (N(2) with <2% residual O(2), in both compartments), 2) hypercapnic anoxia (N(2) with 30% CO(2) and <2% residual O(2), in both compartments), and 3) a 2-phase approach with a hypercapnic hyperoxygenated anesthetic phase (40% CO(2), 30% O(2), and 30% N(2), in the first compartment, 80 s) followed by a second euthanasia phase (80% CO(2) in air, in the second compartment). All 3 CAS approaches effectively achieved nonrecovery states, and time to loss of consciousness for each bird was determined by visual determination of isoelectric EEG and by calculation of the correlation dimension of the EEG. Hypercapnic anoxia resulted in rapid unconsciousness and death; both anoxic treatments were associated with early onset prolonged wing flapping and sustained tonic convulsions as displayed in the electrophysiological recordings. These responses were seen in the period when consciousness remained a possibility. Hypercapnic hyperoxygenation (the 2-phase approach) was associated with respiratory disruption, but this treatment eliminated initial clonic convulsions in the stunning process, and tonic convulsions were not seen. These results suggest that the presence of O(2) in the first stage of CAS is associated with an absence of potentially distressing behavioral responses. The respiratory discomfort associated with hypercapnic hyperoxygenation is an issue. We propose that this may be compensated by a more gradual induction to unconsciousness, which eliminates the impact of other potentially negative experiences.

Spontaneous and odour evoked activity in single avian olfactory bulb neurones.

Extracellular single unit recordings were made from various depths in the left olfactory bulb of ten anaesthetised, freely breathing adult hens (Gallus domesticus) using glass insulated tungsten microelectrodes. The 66 spontaneously active neurons recorded had widely variable firing rates (mean 4.9 spikes/s, range 0.1-32.4 spikes/s) and variable temporal firing patterns (regular, bursting and random discharge). Interspike interval histograms were constructed for each unit and tested for goodness of fit to theoretical distributions. The activity of 23 units fitted gamma distributions, six units fitted lognormal distributions, three units fitted Weibull distributions, one unit fitted an exponential distribution and 11 units had bimodal distributions. Responses of some units to odour stimuli (Clove oil, Geraniol, Limonene and Ammonia all at 10% vapour saturation) delivered directly to the olfactory epithelium were examined. Odour stimulation modified spontaneous activity in 29 of the 44 units tested, causing either inhibition (56%) or excitation (44%) of firing. Of the 35 units stimulated with more than one odour, 15 responded to multiple stimuli, showing excitation or inhibition only or both inhibition and excitation, depending on the odour applied. Ammonia most commonly elicited a response (70% of applications) and all the odours were capable of inducing both inhibition and excitation of spontaneous firing. Examination of transverse sections of adult hen olfactory bulb in the region corresponding to the recording sites allowed unit recording depth to be related to cellular layers. There was some evidence of different neurone properties in each bulb layer. The spontaneous activity and odour responses observed resemble those seen in other vertebrate groups and the firing rates observed are between those previously reported for mammals and reptiles. This study provides detailed physiological evidence for a well-developed functional olfactory system in an avian species.

Survey of the frequency and perceived stressfulness of ethical dilemmas encountered in UK veterinary practice.

The scale of the ethical challenges faced by veterinary surgeons and their perceived stressful consequences were investigated via a short questionnaire, completed by 58 practising veterinary surgeons. Respondents were asked to report how frequently they faced ethical dilemmas, and to rate on a simple numerical scale (zero to 10) how stressful they found three common scenarios. Fifty seven per cent of respondents reported that they faced one to two dilemmas per week, while 34 per cent stated they typically faced three to five dilemmas per week. The three scenarios provided were all rated as highly stressful with ‘client wishing to continue treatment despite poor animal welfare’ rated as the most stressful (median 9). The female veterinary surgeons gave two of the scenarios significantly higher stress ratings than the male veterinary surgeons. Stress ratings were not influenced by number of years in practice (which ranged from one to more than 25 years). The results show that veterinary surgeons regularly face ethical dilemmas and that they find these stressful. This has implications for the wellbeing of veterinary surgeons and supports the case for increased provision of training and support, especially for those who entered the profession before undergraduate ethics teaching was widely available.

Controlled atmosphere stunning of broiler chickens. I. Effects on behaviour, physiology and meat quality in a pilot scale system at a processing plant

1. The effects of controlled atmosphere stunning on the behaviour, physiology and carcase and meat quality of broiler chickens were studied experimentally in a pilot scale plant. 2. Gas mixtures tested were: single phase anoxic mixture (90% Ar in air, <2% O2); single phase hypercapnic anoxic mixture (60% Ar, 30% CO2 in air, <2% O2); and biphasic hypercapnic hyperoxygenation mixture (anaesthetic phase, 40% CO2, 30% O2, 30% N2; euthanasia phase, 80% CO2, 5% O2, 15% N2). 3. Anoxic stunning resulted in the least respiratory disruption, mandibulation and motionlessness, but most head shaking, leg paddling and twitching. Loss of posture occurred soonest with hypercapnic anoxia with the earliest and most twitching and wing flapping in individuals and earliest leg paddling. Biphasic birds were most alert, exhibited most respiratory disruption and mandibulation, and had the latest loss of posture and fewest, but longest bouts of wing flapping and least leg paddling and twitching. 4. Significant and sudden bradycardia and arrhythmia were evident with all gas mixtures and were not related solely to anoxia or hypercapnia. Birds stunned by Ar anoxia showed a slightly more gradual decline from baseline rates, compared with hypercapnic mixtures. 5. Few differences were found between gas mixes in terms of carcase and meat quality. Initial bleeding rate was slowest in biphasic-stunned birds, but total blood loss was not affected. Acceleration of post-mortem metabolism in anoxic-stunned birds was not sufficient to allow de-boning within 5 h without the risk of tough meat. 6. On welfare grounds and taking into account other laboratory and field studies, a biphasic method (using consecutive phases of anaesthesia and euthanasia) of controlled atmosphere stunning of broilers is potentially more humane than anoxic or hypercapnic anoxic methods using argon or nitrogen.

Controlled atmosphere stunning of broiler chickens. II. Effects on behaviour, physiology and meat quality in a commercial processing plant

1. The effects of controlled atmosphere stunning on behavioural and physiological responses, and carcase and meat quality of broiler chickens were studied experimentally in a full scale processing plant. 2. The gas mixtures tested were a single phase hypercapnic anoxic mixture of 60% Ar and 30% CO2 in air with <2% O2, and a biphasic hypercapnic hyperoxygenation mixture, comprising an anaesthetic phase, 40% CO2, 30% O2, 30% N2, followed by an euthanasia phase, 80% CO2, 5% O2, 15% N2. 3. Birds stunned with Ar + CO2 were more often observed to flap their wings earlier, jump, paddle their legs, twitch and lie dorsally (rather than ventrally) than those stunned with CO2 + O2. These behaviours indicate a more agitated response with more severe convulsions during hypercapnic anoxia, thereby introducing greater potential for injury. 4. Heart rate during the first 100 s of gas stunning was similar for both gases, after which it remained constant at ≈230 beats/min for CO2 + O2 birds whereas it declined gently for Ar + CO2 birds. 5. In terms of carcase and meat quality, there appeared to be clear advantages to the processor in using CO2 + O2 rather than Ar + CO2 to stun broiler chickens, for example, a much smaller number of fractured wings (1·6 vs. 6·8%) with fewer haemorrhages of the fillet. 6. This study supports the conclusions of both laboratory and pilot scale experiments that controlled atmosphere stunning of broiler chickens based upon a biphasic hypercapnic hyperoxygenation approach has advantages, in terms of welfare and carcase and meat quality, over a single phase hypercapnic anoxic approach employing 60% Ar and 30% CO2 in air with <2% O2.

Evaluation of the effects of infrared beak trimming in broiler breeder chicks.

Detailed behavioural observations were made of broiler breeder chicks after their beaks had been trimmed by an automated infrared treatment at one day of age or by the traditional hot-blade method at one day or seven days, or after they had been sham-trimmed or left untreated. Observations took place immediately after the treatments and at regular intervals until six weeks of age; beak length and bodyweight were also measured regularly. There were no significant effects on the behaviour of the chicks in the first hour after trimming or in the subsequent six weeks. Variability in beak length was low within the treatments and there was significant regrowth, but it was least in the birds that had been hot-blade trimmed at seven days. Both beak-trimming methods were associated with small but significant reductions in bodyweight, with the hot-blade-treated birds being more affected.

Stimulus-response functions of single avian olfactory bulb neurones.

This study investigated olfactory processing in a functional context by examining the responses of single avian olfactory bulb neurones to two biologically important gases over relevant concentration ranges. Recordings of extracellular spike activity were made from 80 single units in the left olfactory bulb of 11 anaesthetised, freely breathing adult hens (Gallus domesticus). The units were spontaneously active, exhibiting widely variable firing rates (0.07-47.28 spikes/s) and variable temporal firing patterns. Single units were tested for their response to an ascending concentration series of either ammonia (2.5-100 ppm) or hydrogen sulphide (1-50 ppm), delivered directly to the olfactory epithelium. Stimulation with a calibrated gas delivery system resulted in modification of spontaneous activity causing either inhibition (47% of units) or excitation (53%) of firing. For ammonia, 20 of the 35 units tested exhibited a response, while for hydrogen sulphide, 25 of the 45 units tested were responsive. Approximate response thresholds for ammonia (median threshold 3.75 ppm (range 2.5-60 ppm, n=20)) and hydrogen sulphide (median threshold 1 ppm (range 1-10 ppm, n=25)) were determined with most units exhibiting thresholds near the lower end of these ranges. Stimulus response curves were constructed for 23 units; 16 (the most complete) were subjected to a linear regression analysis to determine whether they were best fitted by a linear, log or power function. No single function provided the best fit for all the curves (seven were linear, eight were log, one was power). These findings show that avian units respond to changes in stimulus concentration in a manner generally consistent with reported responses in mammalian olfactory bulb neurones. However, this study illustrates a level of fine-tuning to small step changes in concentration (<5 ppm) not previously demonstrated in vertebrate single olfactory bulb neurones.

Mechanothermal nociceptors in the scaly skin of the chicken leg

Electrophysiological recordings were made from single sensory mechanothermal nociceptive afferent fibres in dissected nerve filaments of the parafibular nerve innervating the scaly skin of the lower leg of the chicken. Two classes of mechanothermal nociceptors were identified consisting of 34 C fibres (conduction velocities 0.45-1.5 m/s, mean 1.08) and nine A-delta fibres (3-15 m/s, mean 6.34). The C fibre afferents had receptive fields which were circular or elliptical in shape and ranged in size from 1 mm in diameter to 4 x 3 mm. Thresholds to mechanical stimulation in the C fibre afferents ranged from 0.3 to 33 g (median 1.5 g) and thermal thresholds were in the range 39-61 degrees C (median 49.4 degrees C). Stimulus-response curves to thermal and/or mechanical stimulation were recorded from 28 C fibre afferents and subjected to a linear regression analysis to determine whether they were best fitted by a linear, log or power function. The results were variable and no single function provided the best fit for all the responses. Of the fibres tested with both stimulus modalities (n=17), only 12 fibres showed the same best fit for both stimuli; in the others the best fit regression lines differed between stimuli. The response of the A-delta fibres to mechanical and thermal stimulation was very similar to the C fibres but the small number of A-delta fibres precluded any detailed statistical analysis. Comparison of the physiological properties of the C fibres in the leg with those previously identified in the beak showed that those in the leg had significantly lower thermal thresholds, but higher mechanical thresholds. The possible functional significance of these differences is discussed. These findings are also discussed in a comparative context to identify similarities and differences between mechanothermal nociceptors in birds and other vertebrates, relating these to their possible evolutionary and functional significance.

Response characteristics of nasal trigeminal nociceptors in Gallus domesticus

Although the chemoreceptive properties of the nasal trigeminal system are well known, the physiological characteristics of nasal nociceptors responding to both mechanical and chemical stimulation have not been well described. In this study, the activity of single nasal trigeminal units recorded from microdissected twigs of the ethmoidal nerve of anaesthetized hens (Gallus domesticus) was investigated. Using a mechanical search stimulus, 20 slowly and 22 rapidly adapting nasal mechanoreceptors were identified, exhibiting mean thresholds of 2.96 g. Twelve slowly adapting units also exhibited chemical sensitivity when exposed to ammonia gas. These had mean response thresholds of 0.232% vapour saturation and exhibited variable stimulus-response profiles. This is the first study to quantify the responses of polymodal nasal nociceptors to a noxious airborne chemical in any species.

Skin temperature reveals the intensity of acute stress.

Acute stress triggers peripheral vasoconstriction, causing a rapid, short-term drop in skin temperature in homeotherms. We tested, for the first time, whether this response has the potential to quantify stress, by exhibiting proportionality with stressor intensity. We used established behavioural and hormonal markers: activity level and corticosterone level, to validate a mild and more severe form of an acute restraint stressor in hens (Gallus gallus domesticus). We then used infrared thermography (IRT) to non-invasively collect continuous temperature measurements following exposure to these two intensities of acute handling stress. In the comb and wattle, two skin regions with a known thermoregulatory role, stressor intensity predicted the extent of initial skin cooling, and also the occurrence of a more delayed skin warming, providing two opportunities to quantify stress. With the present, cost-effective availability of IRT technology, this non-invasive and continuous method of stress assessment in unrestrained animals has the potential to become common practice in pure and applied research.