Behavioural repertoire of working donkeys and consistency of behaviour over time, as a preliminary step towards identifying pain-related behaviours.

BACKGROUND: The donkey has a reputation for stoicism and its behavioural repertoire in clinical contexts is under-reported. Lack of understanding of the norms of donkey behaviour and how it may vary over time can compromise use of behavioural measures as indicators of pain or emotional state. The objective of this study was to find out whether the behaviour of working donkeys was influenced by gender, the time of day or differed between days with a view to assessing how robust these measures are for inclusion in a working donkey ethogram. METHODOLOGY/PRINCIPAL FINDINGS: Frequency and consistency of postural and event behaviours were measured in 21 adult working donkeys (12 females; 9 males). Instantaneous (scan) and focal sampling were used to measure maintenance, lying, ingestive and investigative behaviours at hourly intervals for ten sessions on each of two consecutive days. High head carriage and biting were seen more frequently in male donkeys than females (P<0.001). Level head carriage, licking/chewing and head-shaking were observed more frequently in female donkeys (P<0.001). Tail position, ear orientation, foot stamping, rolling/lying and head-shaking behaviours were affected by time of day (P<0.001). However, only two variations in ear orientation were found to be significantly different over the two days of observations (P<0.001). Tail swishing, head shaking, foot stamping, and ears held sideways and downwards were significantly correlated (P<0.001) and are assumed to be behaviours to discourage flies. CONCLUSIONS/SIGNIFICANCE: All donkeys expressed an extensive behavioural repertoire, although some differences in behaviour were evident between genders. While most behaviours were consistent over time, some behaviours were influenced by time of day. Few behaviours differed between the two test days. The findings can be used to inform the development of a robust, evidence-based ethogram for working donkeys.

Thermal nociceptive threshold testing detects altered sensory processing in broiler chickens with spontaneous lameness.

Lameness is common in commercially reared broiler chickens but relationships between lameness and pain (and thus bird welfare) have proved complex, partly because lameness is often partially confounded with factors such as bodyweight, sex and pathology. Thermal nociceptive threshold (TNT) testing explores the neural processing of noxious stimuli, and so can contribute to our understanding of pain. Using an acute model of experimentally induced articular pain, we recently demonstrated that TNT was reduced in lame broiler chickens, and was subsequently attenuated by administration of Non-Steroidal Anti-Inflammatory Drugs (NSAIDs). This study extended these findings to a large sample of commercial broilers. It examined factors affecting thermal threshold (Part 1) and the effect of an NSAID drug (meloxicam, 5 mg/kg) and of an opioid (butorphanol; 4 mg/kg) (Part 2). Spontaneously lame and matched non-lame birds (n=167) from commercial farms were exposed to ramped thermal stimulations via a probe attached to the lateral aspect of the tarsometatarsus. Baseline skin temperature and temperature at which a behavioural avoidance response occurred (threshold) were recorded. In Part 1 bird characteristics influencing threshold were modelled; In Part 2 the effect of subcutaneous administration of meloxicam or butorphanol was investigated. Unexpectedly, after accounting for other influences, lameness increased threshold significantly (Part 1). In Part 2, meloxicam affected threshold differentially: it increased further in lame birds and decreased in non-lame birds. No effect of butorphanol was detected. Baseline skin temperature was also consistently a significant predictor of threshold. Overall, lameness significantly influenced threshold after other bird characteristics were taken into account. This, and a differential effect of meloxicam on lame birds, suggests that nociceptive processing may be altered in lame birds, though mechanisms for this require further investigation.

Thermal nociception as a measure of non-steroidal anti-inflammatory drug effectiveness in broiler chickens with articular pain.

Pain associated with poultry lameness is poorly understood. The anti-nociceptive properties of two non-steroidal anti-inflammatory drugs (NSAIDs) were evaluated using threshold testing in combination with an acute inflammatory arthropathy model. Broilers were tested in six groups (n=8 per group). Each group underwent a treatment (saline, meloxicam (3 or 5mg/kg) or carprofen (15 or 25mg/kg)) and a procedure (Induced (arthropathy-induction) or sham (sham-handling)) prior to testing. Induced groups had Freund's complete adjuvant injected intra-articularly into the left intertarsal joint (hock). A ramped thermal stimulus (1°C/s) was applied to the skin of the left metatarsal. Data were analysed using random-intercept multi-level models. Saline-induced birds had a significantly higher skin temperature (± SD) than saline-sham birds (37.6 ± 0.8°C vs. 36.5 ± 0.5°C; Z=-3.47, P<0.001), consistent with an inflammatory response. Saline was associated with significantly lower thermal thresholds (TT) than analgesic treatment (meloxicam: Z=2.72, P=0.007; carprofen: Z=2.58, P=0.010) in induced birds. Saline-induced birds also had significantly lower TT than saline-sham birds (Z=-2.17, P=0.030). This study found direct evidence of an association between inflammatory arthropathies and thermal hyperalgesia, and showed that NSAID treatment maintained baseline thermal sensitivity (via anti-nociception). Quantification of nociceptive responsiveness in a predictable broiler pain model identified thermal anti-hyperalgesic properties of two NSAIDs, which suggested that therapeutically effective treatment was provided at the doses administered. Such validation of analgesic strategies will increase the understanding of pain associated with specific natural broiler lameness types.

Kinematic analysis quantifies gait abnormalities associated with lameness in broiler chickens and identifies evolutionary gait differences.

This is the first time that gait characteristics of broiler (meat) chickens have been compared with their progenitor, jungle fowl, and the first kinematic study to report a link between broiler gait parameters and defined lameness scores. A commercial motion-capturing system recorded three-dimensional temporospatial information during walking. The hypothesis was that the gait characteristics of non-lame broilers (n = 10) would be intermediate to those of lame broilers (n = 12) and jungle fowl (n = 10, tested at two ages: immature and adult). Data analysed using multi-level models, to define an extensive range of baseline gait parameters, revealed inter-group similarities and differences. Natural selection is likely to have made jungle fowl walking gait highly efficient. Modern broiler chickens possess an unbalanced body conformation due to intense genetic selection for additional breast muscle (pectoral hypertrophy) and whole body mass. Together with rapid growth, this promotes compensatory gait adaptations to minimise energy expenditure and triggers high lameness prevalence within commercial flocks; lameness creating further disruption to the gait cycle and being an important welfare issue. Clear differences were observed between the two lines (short stance phase, little double-support, low leg lift, and little back displacement in adult jungle fowl; much double-support, high leg lift, and substantial vertical back movement in sound broilers) presumably related to mass and body conformation. Similarities included stride length and duration. Additional modifications were also identified in lame broilers (short stride length and duration, substantial lateral back movement, reduced velocity) presumably linked to musculo-skeletal abnormalities. Reduced walking velocity suggests an attempt to minimise skeletal stress and/or discomfort, while a shorter stride length and time, together with longer stance and double-support phases, are associated with instability. We envisage a key future role for this highly quantitative methodology in pain assessment (associated with broiler lameness) including experimental examination of therapeutic agent efficacy.

A comparison between pre-operative carprofen and a long-acting sufentanil formulation for analgesia after ovariohysterectomy in dogs.

OBJECTIVE: To assess the analgesic efficacy and adverse effects of a novel, long-acting sufentanil preparation in dogs undergoing ovariohysterectomy (OHE). STUDY DESIGN: Blinded, positively controlled, randomized field trial with four parallel treatment groups. ANIMALS: Eighty client owned dogs undergoing elective OHE randomly allocated into four treatment groups (each n = 20). MATERIALS AND METHODS: Three groups received intramuscular (IM) sufentanil (at 10, 15 and 25 microg kg(-1), respectively) and the control group received subcutaneous (SC) carprofen 4 mg kg(-1) SC plus acepromazine 0.05 mg kg(-1) IM as pre-anaesthetic medication. OHE was performed under thiopental/halothane anaesthesia. Visual Analogue Scale (VAS) scores for pain and sedation were awarded and mechanical nociceptive thresholds were measured at the wound and hock before surgery and up to 24 hours after tracheal extubation. Serum cortisol was measured before surgery, during surgery and up to 24 hours after tracheal extubation. Animals with inadequate post-operative analgesia were given rescue medication. RESULTS: In the carprofen group, VAS pain scores were significantly higher, wound tenderness was greater and requirement for rescue analgesia was more than in the sufentanil-treated groups. Sufentanil produced dose dependent analgesia and sedation. All treatment groups showed similar patterns of change for cortisol concentrations. Use of the sufentanil preparation was associated with a relatively high incidence of adverse events. CONCLUSIONS: The long-acting preparation of sufentanil provided excellent post-operative analgesia that was significantly better than that provided by carprofen. However, use of this formulation, in the anaesthetic technique used in the study, resulted in a relatively high incidence of adverse effects. CLINICAL RELEVANCE: Full mu (MOP) opioid agonists provide significantly better post-operative analgesia than nonsteroidal anti-inflammatory drugs after moderately painful surgery. However, the widely recognized adverse effects of opioids may preclude the use of these agents.

Assessment of the influence of surgical technique on postoperative pain and wound tenderness in cats following ovariohysterectomy.

Elective ovariohysterectomy was performed on 66 cats. Surgical approach was flank (group F) or midline (group M) allocated by block randomisation. Pre-anaesthetic medication was acepromazine (0.1 mg/kg) via intramuscular injection. Anaesthesia was induced with intravenous thiopentone, and maintained with halothane in 100% oxygen. Carprofen (4 mg/kg) was administered by the subcutaneous route immediately after induction of anaesthesia. Postoperative pain and wound tenderness were assessed at 1, 3, 6, 9, 11-12 and 20-24h after the end of surgery, and the assessment outcome marked on visual analogue scales (VAS). Intervention analgesia (if pain VAS was >40 mm) was pethidine 4 mg/kg via intramuscular injection. Area under the curve (AUC) for VAS for pain and VAS for wound tenderness for each cat were calculated. AUC for wound tenderness was significantly greater for group F (P = 0.007). There was no significant difference for AUC for pain between the groups. In conclusion, wounds after flank ovariohysterectomy are significantly more tender than after midline ovariohysterectomy in the cat. This indicates that interactive methods, including wound palpation, must be used to assess postoperative pain and the findings should be appropriately weighted in the overall assessment.

Investigation of the EEG effects of intravenous lidocaine during halothane anaesthesia in ponies.

OBJECTIVE: To record the electroencephalographic changes during castration in ponies anaesthetized with halothane and given intravenous (IV) lidocaine by infusion. The hypothesis tested was that in ponies, IV lidocaine is antinociceptive and would therefore obtund EEG changes during castration. ANIMALS: Ten Welsh mountain ponies referred to the Department of Clinical Veterinary Medicine, Cambridge for castration under general anaesthesia. MATERIALS AND METHODS: Following pre-anaesthetic medication with intramuscular acepromazine (0.02 mg kg(-1)) anaesthesia was induced with IV guaiphenesin (60 mg kg(-1)) and thiopental (9 mg kg(-1)) and maintained with halothane at an end-tidal concentration (FE'HAL) of 1.2%. A constant rate infusion of IV lidocaine (100 microg kg(-1) minute(-1)) was administered throughout anaesthesia. The electroencephalogram (EEG) was recorded continuously using subcutaneous needle electrodes. All animals were castrated using a closed technique. The raw EEG signal was analysed after completion of each investigation, and the mean values of EEG variables (median frequency, spectral edge frequency, total amplitude) recorded during a baseline period (before surgery began) and the removal of each testicle were compared using anova for repeated measures. RESULTS: Spectral edge frequency (SEF) 95% decreased during removal of the second testicle compared with baseline recordings. No other significant EEG changes during castration were measured. CONCLUSIONS: Lidocaine obtunded the EEG changes identified during castration in a previous control study, providing indirect evidence that lidocaine administered peri-operatively was antinociceptive and contributed to anaesthesia during castration. CLINICAL RELEVANCE: The antinociceptive effect of lidocaine combined with its minimal cardiovascular effects indicate a potential use for systemic lidocaine in clinical anaesthetic techniques.

Changes in the EEG during castration in horses and ponies anaesthetized with halothane.

OBJECTIVE: To identify changes in the amplitude spectrum of the electroencephalogram (EEG) during a standardized surgical model of nociception in horses. ANIMALS: Thirteen entire male horses and ponies referred to Division of Clinical Veterinary Science, Bristol (n = 9) and Department of Clinical Veterinary Medicine (n = 4) for castration. MATERIALS AND METHODS: Following pre-anaesthetic medication with acepromazine, anaesthesia was induced with guaiphenesin and thiopental and maintained with halothane in oxygen. The EEG was recorded continuously using subcutaneous needle electrodes. Additional monitoring comprised ECG, arterial blood pressure, blood gas analysis, airway gases, and body temperature. All animals were castrated using a closed technique. The raw EEG was analysed after completion of each investigation and the EEG variables median frequency (F50), spectral edge frequency (SEF) 95% and total amplitude were derived from the spectra using standard techniques. The mean values of EEG variables recorded during a baseline time period (recorded before the start of surgery) and castration of each testicle were compared using analysis of variance for repeated measures. RESULTS: Total amplitude (Atot) decreased and F50 increased during castration of each testicle compared to the baseline time period [(89.0 +/- 7.8% testicle 1, 87.0 +/- 7.8% testicle 2) and (110.0 +/- 15.0% testicle 1, 109.0 +/- 15.0% testicle 2), respectively]. Changes in SEF 95% were not significant. CONCLUSIONS: De-synchronization was identified in the EEG during the nociceptive stimulus of castration. The results suggest that an increase in F50 may be a specific marker for nociception in the horse. CLINICAL RELEVANCE: Studies investigating the efficacy of analgesic agents in horses are limited by difficulties in peri-operative pain assessment. This model, using EEG changes associated with nociceptive stimulation, can be used to investigate the anti-nociceptive efficacy of different anaesthetic agents in the horse.