Sandercock Da

Avian reflex and electroencephalogram responses in different states of consciousness.

Defining states of clinical consciousness in animals is important in veterinary anaesthesia and in studies of euthanasia and welfare assessment at slaughter. The aim of this study was to validate readily observable reflex responses in relation to different conscious states, as confirmed by EEG analysis, in two species of birds under laboratory conditions (35-week-old layer hens (n=12) and 11-week-old turkeys (n=10)). We evaluated clinical reflexes and characterised electroencephalograph (EEG) activity (as a measure of brain function) using spectral analyses in four different clinical states of consciousness: conscious (fully awake), semi-conscious (sedated), unconscious-optimal (general anaesthesia), unconscious-sub optimal (deep hypnotic state), as well as assessment immediately following euthanasia. Jaw or neck muscle tone was the most reliable reflex measure distinguishing between conscious and unconscious states. Pupillary reflex was consistently observed until respiratory arrest. Nictitating membrane reflex persisted for a short time (<1 min) after respiratory arrest and brain death (isoelectric EEG). The results confirm that the nictitating membrane reflex is a conservative measure of death in poultry. Using spectral analyses of the EEG waveforms it was possible to readily distinguish between the different states of clinical consciousness. In all cases, when birds progressed from a conscious to unconscious state; total spectral power (PTOT) significantly increased, whereas median (F50) and spectral edge (F95) frequencies significantly decreased. This study demonstrates that EEG analysis can differentiate between clinical states (and loss of brain function at death) in birds and provides a unique integration of reflex responses and EEG activity.

Histopathological characterization of tail injury and traumatic neuroma development after tail docking in piglets

Summary Tail docking of neonatal pigs is widely used as a measure to reduce the incidence of tail biting, a complex management problem in the pig industry. Concerns exist over the long-term consequences of tail docking for possible tail stump pain sensitivity due to the development of traumatic neuromas in injured peripheral nerves. Tail stumps were obtained post mortem from four female pigs at each of 1, 4, 8 and 16 weeks following tail amputation (approximately two-thirds removed) by a gas-heated docking iron on post natal day 3. Tissues were processed routinely for histopathological examination. Non-neural inflammatory and reparative epidermal and dermal changes associated with tissue thickening and healing were observed 1 to 4 months after docking. Mild neutrophilic inflammation was present in some cases, although this and other degenerative and non-neural reparative changes are not likely to have caused pain. Traumatic neuroma and neuromatous tissue development was not observed 1 week after tail docking, but was evident 1 month after tail docking. Over time there was marked nerve sheath and axonal proliferation leading to the formation of neuromata, which were either localized and circumscribed or comprised of multiple axons dispersed within granulation tissue. Four months after tail resection, neuroma formation was still incomplete, with possible implications for sensitivity of the tail stump.

Current and novel methods for killing poultry individually on-farm

This review examines methods for culling small numbers of poultry on farm, considering both common techniques and methods that are yet to be tested on poultry. The aim of this review is to inform the design of experiments that will assess the pros and cons of culling techniques. The methods reviewed include manual and mechanical cervical dislocation, crushing methods (such as burdizzos or pliers), percussive devices, blunt force trauma and a brain-stem piercing device. Previous work on these approaches, of which there is a limited dataset, has relied on behavioural and brain activity as proxy measures of unconsciousness (insensibility) and death, however there remains some uncertainty as to when birds can be considered unequivocally insensible. These factors will be considered when deciding which of the methods will be recommended to be taken forward for further assessment.

Characterization of short- and long-term mechanical sensitisation following surgical tail amputation in pigs

Commercial pigs are frequently exposed to tail mutilations in the form of preventive husbandry procedures (tail docking) or as a result of abnormal behaviour (tail biting). Although tissue and nerve injuries are well-described causes of pain hypersensitivity in humans and in rodent animal models, there is no information on the changes in local pain sensitivity induced by tail injuries in pigs. To determine the temporal profile of sensitisation, pigs were exposed to surgical tail resections and mechanical nociceptive thresholds (MNT) were measured in the acute (one week post-operatively) and in the long-term (either eight or sixteen weeks post-surgery) phase of recovery. The influence of the degree of amputation on MNTs was also evaluated by comparing three different tail-resection treatments (intact, ‘short tail’, ‘long tail’). A significant reduction in MNTs one week following surgery suggests the occurrence of acute sensitisation. Long-term hypersensitivity was also observed in tail-resected pigs at either two or four months following surgery. Tail amputation in pigs appears to evoke acute and sustained changes in peripheral mechanical sensitivity, which resemble features of neuropathic pain reported in humans and other species and provides new information on implications for the welfare of animals subjected to this type of injury.

Application of a handheld Pressure Application Measurement device for the characterisation of mechanical nociceptive thresholds in intact pig tails.

The assessment of nociceptive thresholds is employed in animals and humans to evaluate changes in sensitivity potentially arising from tissue damage. Its application on the intact pig tail might represent a suitable method to assess changes in nociceptive thresholds arising from tail injury, such as tail docking or tail biting. The Pressure Application Measurement (PAM) device is used here for the first time on the tail of pigs to determine the reliability of the methods and to provide novel data on mechanical nociceptive thresholds (MNT) associated with four different age groups (9, 17, 24 and 32 weeks) and with proximity of the target region to the body of the animal. We recorded an overall acceptable level of intra-individual reliability, with mean values of CV ranging between 30.1 and 32.6%. Across all age groups, the first single measurement of MNT recorded at region 1 (proximal) was significantly higher (P < 0.05) than the following two. This was not observed at tail regions 2 and 3 (more distal). Age had a significant effect (P < 0.05) on the mean thresholds of nociception with increasing age corresponding to higher thresholds. Furthermore, a significant effect of proximity of tail region to the body was observed (P < 0.05), with MNT being higher in the proximal tail region in pigs of 9, 17 and 24 weeks of age. There was also a significant positive correlation (P < 0.05) between mechanical nociceptive thresholds and age/body size of the animals. To the best of our knowledge, no other investigation of tail nociceptive thresholds has been performed with the PAM device or alternative methods to obtain mechanical nociceptive thresholds in intact tails of pigs of different age/body size. The reliability of the data obtained with the PAM device support its use in the measurement of mechanical nociceptive threshold in pig tails. This methodological approach is possibly suitable for assessing changes in tail stump MNTs after tail injury caused by tail docking and biting.

Welfare Risks of Repeated Application of On-Farm Killing Methods for Poultry

Simple Summary During poultry production, some birds are killed humanely on farm, usually because they are ill or injured. Recent European Union (EU) legislation has restricted the number of birds that can be killed by manual neck dislocation to 70 birds per person per day. We examined whether this limit is meaningful by investigating the effects of repeated application of two methods of killing (neck dislocation and a percussive method, the CashPoultry Killer). Twelve male stockworkers each killed 100 birds (broilers, laying hens, or turkeys) at a fixed rate with each method. Both methods were highly successful, and reflex and behaviour measures confirmed they caused rapid loss of brain function. Importantly, there was no evidence of reduced performance with time/bird number up to 100 birds with either method. The Cash Poultry Killer caused a more rapid death, but it was prone to technical difficulties with repeated use. Neck dislocation has the important advantage that it can be performed immediately with no equipment, which may make it preferable in some situations. We present the first evidence that, at the killing rates tested, there was no evidence to justify the current EU number limit for performance of neck dislocation to kill poultry on farm. Abstract Council Regulation (EC) no. 1099/2009 on the protection of animals at the time of killing restricts the use of manual cervical dislocation in poultry on farms in the European Union (EU) to birds weighing up to 3 kg and 70 birds per person per day. However, few studies have examined whether repeated application of manual cervical dislocation has welfare implications and whether these are dependent on individual operator skill or susceptibility to fatigue. We investigated the effects of repeated application (100 birds at a fixed killing rate of 1 bird per 2 min) and multiple operators on two methods of killing of broilers, laying hens, and turkeys in commercial settings. We compared the efficacy and welfare impact of repeated application of cervical dislocation and a percussive killer (Cash Poultry Killer, CPK), using 12 male stockworkers on three farms (one farm per bird type). Both methods achieved over 96% kill success at the first attempt. The killing methods were equally effective for each bird type and there was no evidence of reduced performance with time and/or bird number. Both methods of killing caused a rapid loss of reflexes, indicating loss of brain function. There was more variation in reflex durations and post-mortem damage in birds killed by cervical dislocation than that found using CPK. High neck dislocation was associated with improved kill success and more rapid loss of reflexes. The CPK caused damage to multiple brain areas with little variation. Overall, the CPK was associated with faster abolition of reflexes, with fewer birds exhibiting them at all, suggestive of better welfare outcomes. However, technical difficulties with the CPK highlighted the advantages of cervical dislocation, which can be performed immediately with no equipment. At the killing rates tested, we did not find evidence to justify the current EU limit on the number of birds that one operator can kill on–farm by manual cervical dislocation.

Determination of stable reference genes for RT-qPCR expression data in mechanistic pain studies on pig dorsal root ganglia and spinal cord

RNA expression levels for genes of interest must be normalised with appropriate reference or “housekeeping” genes that are stably expressed across samples and treatments. This study determined the most stable reference genes from a panel of 6 porcine candidate genes: beta actin (ACTB), beta-2-microglobulin (B2M), eukaryotic elongation factor 1 gamma-like protein (eEF-1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), succinate dehydrogenase complex subunit A (SDHA), Ubiquitin C (UBC) in sacral dorsal root ganglia and spinal cord samples collected from 16 tail docked pigs (2/3rds of tail amputated) 1, 4, 8 and 16 weeks after tail injury (4 pigs/time point). Total RNA from pooled samples was measured by SYBRgreen real-time quantitative PCR. Cycle threshold values were analysed using geNorm, BestKeeper and NormFinder PCR analysis software. Average expression stability and pairwise variation values were calculated for each candidate reference gene. GeNorm analysis identified the most stable genes for normalisation of gene expression data to be GAPDH > eEF-1 > UBC > B2M > ACTB > SDHA for dorsal root ganglia and ACTB > SDHA > UBC > B2M > GAPDH > eEF-1 for spinal cord samples. Expression stability estimates were verified by BestKeeper and NormFinder analysis. Expression stability varied between genes within and between tissues. Validation of most stably expressed reference genes was performed by normalisation of calcitonin gene related polypeptide beta (CALCB). The results show similar patterns of CALCB expression when the best reference genes selected by all three programs were used. GAPDH, eEF-1 and UBC are suitable reference genes for porcine dorsal root ganglia samples, whereas ACTB, SDHA and UBC are more appropriate for spinal cord samples.

The effect of tail docking in neonatal pigs on the central expression of genes involved in modulating anxiety-like behaviour

Abstract Background Adverse experiences in early life, such as exposure to stress, can have long term detrimental effects on the future physiology and behaviour of the animal. Typically animals exposed to such experiences are more anxious and more reactive to stress in later life. Tail biting is a major problem in modern pig production, both in terms of animal welfare and productivity. Tail docking in early postnatal life is common practice to reduce risk of this problem, but causes pain and may alter pain sensitivity. Aims To investigate whether a significant painful experience in early life (tail docking) alters the expression of genes in the amygdala that are linked to an anxiety-prone phenotype. Methods Eight female piglets (Landrace/Large White × synthetic sireline) were used. Four piglets were tail docked (amputation of approx. 2/3 of the tail) on post-natal day 3 using hot-iron cautery and four sham-docked piglets served as intact controls. On post-natal day 10, pigs were sedated and then euthanized by barbiturate overdose. Brains were removed, the amygdala grossly dissected and frozen on dry ice. 20 μm sections were cut and subsequently processed using in situ hybridisation with radiolabelled probes complementary to corticotropin-releasing hormone receptor-1 (Crhr1) and CRH receptor-2 (Crhr2) mRNA. Results Crhrl mRNA expression was significantly greater in the amygdala of tail-docked piglets compared with the sham-docked animals. There was no significant difference detected in Crhr2 expression in the amygdala between the groups. Conclusion Increased expression of Crhrl in the amygdala is associated with an anxiety-prone phenotype in rats and pigs, thus it is likely that tail docking in early life leads to enhanced anxiety which may have a negative impact on pig welfare. Ongoing experiments will determine whether these central changes in gene expression are long-lasting. [Support: BBSRC/DEFRA, part of ANIWHA ERA-NET initiative].

Can peripheral nerve damage caused by tail docking lead to tail pain later in the life of pigs

Chronic stress is a long-lasting negative emotional state which induces negative consequences on animals’ behavior. This study aimed at assessing whether unpredictable and repeated negative stimuli (URNS) influence feeding behavior in quail, and whether this can be modulated by their emotionality. Two lines of quail divergently selected on their inherent emotionality (low emotionality, STI; high emotionality, LTI) were either daily exposed to URNS or undisturbed from 17 to 40 days of age (n=32 for each line, in each group). During this time, quail were submitted twice to a sequential feeding procedure: they were offered a hypocaloric diet (7% less caloric than the normocaloric diet) on odd days and a hypercaloric diet (7% more caloric) on even days, for 8 days; then, they received a normocaloric diet (metabolizable energy=12.56 MJ) for 3 days. This sequential feeding procedure was used to assess anhedonia and diet preferences thanks to choice tests (hypo vs. hypercaloric diets) performed at the end of each period. Short-term (30 min) and daily intake were also measured each day. Behavioral tests were performed to assess quail’s emotional reactivity. Results showed that URNS enhanced quails’ emotional reactivity, e.g. in the reactivity to human test, disturbed quail came later (P=0.011) and spent less time (P 0.1), but URNS reduced their daily intake during the 2nd period (P<0.05). Motivation for each diet (assessed by their short-term intake) was differently affected by URNS during the 2nd period: STI quail decreased their motivation to eat the hypercaloric diet (P<0.01) whereas LTI increased their motivation to eat the hypocaloric diet (P<0.01). In conclusion, both lines of quail experienced a chronic stress as URNS induced an increase of their emotional reactivity. Interestingly, URNS induced opposite changes in quail’s feeding behavior: LTI disturbed quail seemed to express a short-term compensatory behavior because of their high motivation to eat, whereas STI disturbed quail seemed to be in a devaluation process as shown by their anhedonia and their decrease of daily intake.Farm animals have to adapt to human presence from birth and being handled may lead to fear and stress reactions. It is known that the mother can be used as a postnatal model in the development of young-human relationship. Through her, some information like auditory ones may even be learnt prenatally. We tested this idea in pigs because they communicate a lot by acoustic signals. The hypotheses were that prenatal experience with human voice could modify behavioural reactions to the experienced voice and to an unfamiliar voice expressing different emotions. We worked with 30 pregnant sows from the last month of gestation. Ten sows (treatment A) were submitted to recordings of human voices during handling: vA during positive interactions and vB during negative interactions, twice a day, 5 days a week, for 10 minutes. Ten other sows (treatment B) received the contrary, i.e. vB during positive interactions and vA during negative interactions. Ten last sows (treatment C) received no vocal stimulations during handling sessions. Two days old piglets (36 A, 39 B, 35 C) were submitted to a 5 min choice test between voices vA and vB in a testing pen (2×1 m). Each voice was played back through loudspeakers positioned at each end of the pen. At 15-18 days of age, 20 other piglets from each treatment were tested in the same conditions except that we played back the voice of an unknown person, reading the same text with a joyful or angry intention. In both tests we recorded vocalisations and locomotion. Data were analysed using non parametric statistics (Statview). In both tests, A and B piglets started to move sooner (P 0.05) to be and the time spent (178 s (46s)) close to the loudspeakers did not depend on the treatment (P>0.05). We also found no difference between the time spent close to one loudspeaker or the other, neither for vA versus vB, nor for joyful versus angry intention (P>0.05). The results show that the prenatal experience of human voice reduces postnatal behavioural reactions of stress (vocalisations, latency to move) during the playback human voices. However, it does not seem to induce specific attraction toward human voice, or human emotional intention. Therefore prenatal experience with human voice may be a good way of reducing fear reactions to human voice after birth.

Approaches to culling poultry humanely on farm

Approaches to culling poultry humanely on farm V. SANDILANDS, D. SANDERCOCK, J. SPARREY, D. McKEEGAN, J. HOPKINS, M. RAJ AND N. SPARKS Avian Science Research Centre, SAC Auchincruive, Ayr KA6 5HW, Livetec Systems Ltd, Flitton, Bedford, MK45 5DT, Institute for Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Bearsden Road, G61 1QH, and School Division of Farm Animal Science, School of Clinical Veterinary Science, University of Bristol, Langford, BS40 5DU (E-mail: vicky.sandilands@sac.ec.uk)