The efficacy of electrical stunning of New Zealand rock lobster (Jasus edwardsii) and freshwater crayfish (Paranephrops zealandicus) using the Crustastun™.

Large numbers of decapod crustacea are farmed and harvested globally for human consumption. Growing evidence for the capacity of these animals to feel pain, and therefore to suffer, has led to increased concern for their welfare, including at slaughter. In New Zealand, decapod crustacea are protected by animal welfare legislation. There is a requirement that all farmed or commercially caught animals of these species killed for commercial purposes are first rendered insensible. The aim of this study was to evaluate the efficacy of the Crustastun™, a commercially available bench-top electrical stunner, in two commercially important New Zealand crustacean species; the rock lobster (Jasus edwardsii) and koura (freshwater crayfish [Paranephrops zealandicus]). Animals were anaesthetised via intramuscular injection of lidocaine and instrumented to record the electrical activity of the nervous system, prior to being stunned according to the manufacturer's instructions. Stunning efficacy was determined by analysing neural activity and observing behaviour post stunning. All ten P. zealandicus and three J. edwardsii appeared to be killed outright by the stun. Of the remaining J. edwardsii, six exhibited some degree of muscle tone and/or slow unco-ordinated movements of the limbs or mouthparts after stunning, although there was no recovery of spontaneous or evoked movements. One J. edwardsii was unable to be stunned successfully, likely due to its very large size (1.76 kg). None of the successfully stunned animals showed any evidence of return of awareness in the five minutes following stunning. It was concluded that the Crustastun™ is an acceptable method for killing P. zealandicus and for stunning all but the largest J. edwardsii.

Electroencephalographic responses of anaesthetized pigs (Sus scrofa) to tail docking using clippers or cautery iron performed at 2 or 20 days of age.

OBJECTIVE: To compare electroencephalographic (EEG) responses of pigs to tail docking using clippers or cautery iron, performed at 2 or 20 days of age. STUDY DESIGN: Prospective, randomised controlled experimental study. ANIMALS: A total of 40 Large White x Landrace entire male pigs aged 2 (n=20) or 20 (n=20) days were randomly assigned to undergo tail docking using clippers or cautery iron. METHODS: Anaesthesia was induced and maintained with halothane delivered in oxygen. Following instrumentation, end-tidal halothane concentration was stabilised at 1.0±0.05%, and EEG recording commenced. After a 5 minute baseline period, tail docking was performed and recording continued for additional 10 minutes. EEG data were subjected to Fast Fourier transformation, yielding the summary variables median frequency (F50), 95% spectral edge frequency (F95) and total power (PTOT). Variables recorded during the baseline period were compared with those calculated at consecutive 15 second intervals following tail docking. RESULTS: Following tail docking, F50 decreased briefly but significantly in 2-day-olds, whereas 20-day-olds exhibited a sustained increase in F50 (p<0.05). Immediately after tail docking, F50 was overall lower in 2-day-olds than in 20-day-olds (p<0.05). F95 increased after docking in 20-day-olds docked using clippers (p<0.05) but did not change in 20-day-olds docked using cautery iron or in 2-day-olds docked using either method. Overall, F95 was lower in 2-day-olds than in 20-day-olds from 30 to 60 seconds after docking (p<0.05). PTOT decreased after docking in 20-day-olds (p<0.05) but did not change in 2-day-olds. Overall, PTOT was lower in 2- than in 20-day-olds during baseline and after tail docking (p<0.05). CONCLUSIONS AND CLINICAL RELEVANCE: These data suggest that tail docking using clippers is more acutely painful than docking using cautery iron and that docking within the first days of birth may be less acutely painful than docking at a later age.

Electroencephalographic assessment of oral meloxicam, topical anaesthetic cream and cautery iron for mitigating acute pain in pigs (Sus scrofa) undergoing tail docking.

OBJECTIVE: To evaluate the efficacy of oral meloxicam, topical anaesthetic cream and cautery iron in mitigating acute nociceptive responses of pigs to tail docking. STUDY DESIGN: A prospective, randomized, controlled experimental study. ANIMALS: A total of 40 healthy Large WhitexLandrace pigs aged 21±1 days, weighing 6.1±0.9 kg. METHODS: Pigs were randomly assigned to one of four treatments (n=10 per treatment): CONTROL: docked using clippers without analgesia; MEL: docked using clippers after administration of oral meloxicam; EMLA: docked using clippers after application of topical anaesthetic cream; and CAUT: docked using a cautery iron without analgesia. Anaesthesia was induced and maintained with halothane in oxygen. Following induction, end-tidal halothane was stabilized at 0.95-1.05% and electroencephalograph (EEG) recording commenced. After 5 minutes of baseline data collection, tail docking was performed and recording continued for a further 10 minutes. The EEG summary variables median frequency (F50), 95% spectral edge frequency (F95) and total power (PTOT) were calculated for the baseline period and for consecutive 30-second intervals following docking. RESULTS: Following docking, F50 increased and PTOT decreased significantly in CONTROL and MEL pigs. EMLA pigs exhibited no change in any variable, whilst CAUT pigs exhibited a reduction in PTOT but no change in F50. F50 was higher in control pigs than in EMLA pigs 30-60 seconds after docking (p≤0.01). PTOT was lower in CONTROL than in EMLA pigs 30-90 seconds after docking (p<0.03) and in CAUT pigs 60 seconds after docking (p=0.01). CONCLUSIONS AND CLINICAL RELEVANCE: Prior application of EMLA cream abolished EEG indicators of nociception in pigs docked using clippers. Docking using a cautery iron without analgesia ameliorated EEG indicators of nociception, relative to using clippers without analgesia. Prior administration of EMLA cream or the use of cautery instead of clippers may reduce the acute pain experienced by pigs undergoing tail docking.

Practicability of a Time-Limited Welfare Assessment Protocol for Pasture-Based Dairy Farms, and a Preliminary Assessment of Welfare Outcome Thresholds.

This study assessed a new time-limited protocol developed for pasture-based cows across 23 dairy farms. The process started prior to milking with a questionnaire, followed by an assessment of resources (16 farms only) and behavioural observation of cows at pasture. Remaining animal-based measures were assessed during milking, usually by two assessors (one parlour based and one outside). The protocol proved to be practical and feasible with limited changes needed, except for the assessment of water availability and behaviour. As most cows could access only one water trough, distance between troughs was not a measure of water availability, while the observation of a large numbers of cows at pasture for 30 min resulted in few observations and an uncertain denominator (effective number of observed cows). Further research is needed to determine the best way of assessing water availability and cow behaviour in a time-limited assessment of pasture-based cows. Three animal-based measures (broken tails, dirtiness, and coughing) had mean values higher than the author-determined acceptable thresholds, while <50% of farms met trough cleanliness and track condition targets, and none met the criteria for shelter and shade. This was a sample of farms based on convenience, so more data are required to establish the representativeness of these results. Such testing should involve assessment of the repeatability and reliability of the measures in our protocol.

Evaluating the Behavioural Responses of Healthy Newborn Calves to a Thoracic Squeeze.

A thoracic squeeze has been observed to cause low-vigour neonates of various farmed mammal species, including calves, to enter a state of reduced responsiveness. The removal of the squeeze causes rapid recovery and the expression of normal, healthy behaviours. However, the responses of healthy calves to a thoracic squeeze have not yet been characterized. The responses of 16 healthy newborn calves to a thoracic squeeze are described, along with the effect of the squeeze's application method on the response. Calves aged between 12 and 36 h were subjected to the squeeze using a rope (n = 8) or an inflation cuff (n = 8). In total, 13 of the 16 calves were induced into a state of reduced responsiveness, though neural reflexes persisted in nearly all of them. The squeeze was discontinued for nearly half of those induced before the end of the 10-min period, either due to spontaneous arousal or physiological instability. Both methods of application were equally effective at inducing reduced responsiveness, though responses to the cuff appeared to be more rapid than those to the rope. These findings support previous research on piglets and foals, and suggest that the behavioural responses to a thoracic squeeze are generalised across neonates of precocial farmed mammals; the findings provide a foundation for further research exploring the mechanisms underlying the response and the benefits that its application may bring for the performance of husbandry procedures.

Pharmacokinetics and efficacy of a novel long-acting bupivacaine formulation for cornual nerve block in calves.

Local anesthetics are commonly used in farm animals to provide analgesia for painful procedures but can cause adverse effects at high systemic concentrations. The pharmacokinetics and efficacy of a long-acting sucrose acetate isobutyrate (SAIB) bupivacaine formulation following cornual nerve block in calves were compared to lidocaine. Fourteen calves were randomly assigned to one of the treatment groups (i) 5% Bupivacaine-SAIB (BUP-SAIB), n = 7; or (ii) 2% lidocaine (LID), n = 7. Cornual nerve block was performed, and duration of effective analgesia was evaluated by nociceptive threshold testing using a hand-held pressure algometer. Blood samples were collected at various time points and plasma concentrations were analyzed by HPLC. Pharmacokinetic parameters were calculated using a non-compartmental model. The mechanical nociceptive thresholds showed that the novel formulation could desensitize the skin around the horn bud for 18.77 ± 8.88 h (range 8-36 h), compared to 0.79 ± 0.34 h (range 0.5-1.5 h) with lidocaine. The mean maximum plasma concentration (Cmax) of bupivacaine was 152.03 (SD 37.34) ng/mL and its Tmax was 0.39 (SD 0.13) h. The half-life of elimination was 32.79 ± 11.00 h and the rate of clearance was 0.12 ± 0.03 L h-1. No toxicity signs were seen after treatment in either group. The novel formulation produced long-lasting analgesia of several times greater duration than that produced by lidocaine. This study showed that the safety and efficacy of the SAIB formulation justifies further studies in a larger population of animals.

Characterisation of the Behavioural Effects of a Thoracic Squeeze in Healthy Newborn Piglets.

A thoracic squeeze has been observed to cause both healthy and low vigour neonatal foals to enter a 'less-responsive state', characterised by loss of posture, eye closure and cessation of movement, from which they rapidly recover to express normal healthy behaviours when the squeeze is released. To date, there have been no systematic studies characterising the responses of healthy neonates of other mammalian species to a thoracic squeeze. We describe the responses of healthy newborn piglets (n = 17) to a standardised application of the thoracic squeeze and evaluate the effect of the method of squeeze application on the response. Neonatal piglets were squeezed around the chest with either a soft fabric rope as has been used in foals (n = 8) or a novel purpose-made inflation cuff (n = 9). Both methods were effective at inducing a less-responsive behavioural state in all piglets, with neural reflexes reduced or absent in over half of them. The inflation cuff appeared to induce the less-responsive state faster than the rope, and more piglets squeezed with the cuff remained in this state for the full 10-min squeeze. These findings suggest that the behavioural response of foals to thoracic squeezing can be generalised to neonates of other precocial mammalian species. This initial study provides a foundation for further research using the inflation cuff to explore mechanisms underlying the thoracic squeeze and ways in which it may be applied whilst performing husbandry procedures.

Animal Welfare Assessment: Can We Develop a Practical, Time-Limited Assessment Protocol for Pasture-Based Dairy Cows in New Zealand?

Despite being a leading producer and exporter of dairy products, New Zealand has no industry-recognised welfare assessment protocol. A New Zealand-specific protocol is essential, as almost all dairy farms in New Zealand are pasture-based and housing is rarely used. Therefore, protocols developed for intensive cows are not suitable. The aim of this study was to develop a simple yet practical welfare assessment protocol that could be used to assess the welfare of a dairy herd during one visit timed to occur around milking. Six welfare assessment protocols and four studies of dairy cattle welfare assessments that had some focus on dairy cattle welfare at pasture were used, along with the New Zealand Dairy Cattle Code of Welfare, to identify potential assessments for inclusion in the protocol. Eighty-four potential assessments (20 record-based and 64 that needed assessing on-farm) were identified by this process of welfare assessments. After screening to exclude on-farm assessments that were not relevant, that had only limited practical application in pasture-based dairy cows or that required more time than available, 28 on-farm assessments remained, which were put together with the 20 record-based assessments and were tested for feasibility, practicality and time on two pasture-based dairy farms. Assessments were then identified as suitable, suitable after modification or not feasible. Suitable and modified assessments were then included in the final protocol alongside additional measures specific to New Zealand dairy farms. The final protocol included 24 on-farm assessments and eight record-based assessments. Further testing of these 32 assessments is needed on more dairy farms across New Zealand before the protocol can be used to routinely assess the welfare of dairy cows in New Zealand.

Risk factors for bobby calf mortality across the New Zealand dairy supply chain.

The objective of this study was to identify risk factors for morbidity and mortality of bobby calves across the whole dairy supply chain in New Zealand. A case-control study was carried out in the 2016 spring calving season. A total of 194 bobby calves, comprising 38 cases (calves that died or were condemned for health or welfare reasons before the point of slaughter) and 156 controls (calves deemed acceptable and presented for slaughter) were included in the study. Case and control calves were selected by veterinarians located at 29 processing premises across New Zealand. Information regarding management of selected calves on-farm, during transport and at the processor was obtained retrospectively via questionnaires administered to supplying farmers, transport operators and processing premises personnel. Associations between management variables and calf mortality (death or condemnation) were examined using multivariable logistic regression models. Factors associated with an increased risk of calf mortality included time in the farm of origin's calving season, duration of travel from farm to the processor and processing slaughter schedule (same day or next day). Every additional week into the farm's calving season increased the odds of mortality by a factor of 1.2 (95%CI 1.06, 1.35). Similarly, each additional hour of travel time increased the odds of mortality by a factor of 1.45 (95% CI 1.18, 1.76). Risk of mortality was significantly greater for calves processed at premises with a next day slaughter schedule than those processed at premises with a same day slaughter schedule (OR 3.82, 95% CI 1.51, 9.67). However, when the data set was limited to those cases that died or were condemned in the yards (i.e. excluding calves that were dead or condemned on arrival) the effect of same day slaughter was not significant. In order to reduce bobby calf mortality and morbidity, transport duration should be kept as short as possible and a same day slaughter schedule applied. While these factors can be regulated, New Zealand's pastoral dairy system means that calves will inevitably be transported for slaughter across several months each spring. Although farm management factors did not apparently influence the risk of mortality in this study, the effect of time in farm's calving season suggests there may be farm-management related factors that change over the season. This requires further investigation.

Evaluation of Different Gases and Gas Combinations for On-Farm Euthanasia of Pre-Weaned Pigs.

The aim of this research was to evaluate the welfare of pre-weaned piglets euthanised using three different gas treatments: 100% carbon dioxide (CO2), 100% argon (Ar) or a mixture of 60% Ar/40% carbon dioxide (Ar/CO2). Two studies (n = 5 piglets/treatment/study) were conducted: (1) behavioural and physiological data were collected from conscious piglets during exposure to test gases via immersion in a pre-filled chamber and (2) electrophysiological data were collected from lightly anaesthetised, intubated and mechanically ventilated piglets exposed to the same test gases. Based on the duration of escape attempts and laboured breathing, piglets exposed to 100% CO2 experienced more stress than piglets exposed to 100% Ar prior to loss of consciousness, but there appeared to be no advantage of mixing Ar with CO2 on indices of animal welfare. However, spectral analysis of the electroencephalogram revealed no changes consistent with nociception during exposure to any of the three gas treatments. Based on the behavioural response to gas exposure, all gases tested caused signs of stress prior to piglets losing consciousness and hence alternative methods of euthanasia need to be evaluated.

Effects of noxious stimuli on the electroencephalogram of anaesthetised chickens (Gallus gallus domesticus).

The reliable assessment and management of avian pain is important in the context of animal welfare. Overtly expressed signs of pain vary substantially between and within species, strains and individuals, limiting the use of behaviour in pain studies. Similarly, physiological indices of pain can also vary and may be confounded by influence from non-painful stimuli. In mammals, changes in the frequency spectrum of the electroencephalogram (EEG) recorded under light anaesthesia (the minimal anaesthesia model; MAM) have been shown to reliably indicate cerebral responses to noxious stimuli in a range of species. The aim of the current study was to determine whether the MAM can be applied to the study of nociception in birds. Ten chickens were lightly anaesthetised with halothane and their EEG recorded using surface electrodes during the application of supramaximal mechanical, thermal and electrical noxious stimuli. Spectral analysis revealed no EEG responses to any of these stimuli. Given that birds possess the neural apparatus to detect and process pain, and that the applied noxious stimuli elicit behavioural signs of pain in conscious chickens, this lack of response probably relates to methodological limitations. Anatomical differences between the avian and mammalian brains, along with a paucity of knowledge regarding specific sites of pain processing in the avian brain, could mean that EEG recorded from the head surface is insensitive to changes in neural activity in the pain processing regions of the avian brain. Future investigations should examine alternative electrode placement sites, based on avian homologues of the mammalian brain regions involved in pain processing.

Effects of halothane on the electroencephalogram of the chicken.

Little is known about the effects of inhalant anaesthetics on the avian electroencephalogram (EEG). The effects of halothane on the avian EEG are of interest, as this agent has been widely used to study nociception and analgesia in mammals. The objective of this study was to characterize the effects of halothane anaesthesia on the EEG of the chicken. Twelve female Hyline Brown chickens aged 8-10 weeks were anaesthetized with halothane in oxygen. For each bird, anaesthesia was progressively increased from 1-1.5 to 2 times the Minimum Anesthetic Concentration (MAC), then progressively decreased again. At each concentration, a sample of EEG was recorded after a 10-min stabilization period. The mean Total Power (PTOT ), Median Frequency (F50) and 95% Spectral Edge Frequency (F95) were calculated at each halothane MAC, along with the Burst Suppression Ratio (BSR). Burst suppression was rare and BSR did not differ between halothane concentrations. Increasing halothane concentration from 1 to 2 MAC resulted in a decrease in F50 and increase in PTOT , while F95 increased when MAC was reduced from 1.5 to 1. The results indicate dose-dependent spectral EEG changes consistent with deepening anaesthesia in response to increasing halothane MAC. As burst suppression was rare, even at 1.5 or 2 times MAC, halothane may be a suitable anaesthetic agent for use in future studies exploring EEG activity in anaesthetized birds.

Nitrous oxide as a humane method for piglet euthanasia: Behavior and electroencephalography (EEG).

The search for humane methods to euthanize piglets is critical to address public concern that current methods are not optimal. Blunt force trauma is considered humane but esthetically objectionable. Carbon dioxide (CO2) is used but criticized as aversive. This research sought to: 1) evaluate the aversiveness of inhaling nitrous oxide (N2O; 'laughing gas') using an approach-avoidance test relying on the piglet's perspective, and 2) validate its humaneness to induce loss of consciousness by electroencephalography (EEG). The gas mixtures tested were N2O and air (90%:10%; '90 N'); N2O, oxygen and air (60%:30%:10%; '60 N'); and CO2 and air (90%:10%; '90 C'). Experiment 1 allowed piglets to walk freely between one chamber filled with air and another prefilled with 60 N or 90 N. All piglets exposed to 60 N lasted for the 10 min test duration whereas all piglets exposed to 90 N had to be removed within 5 min because they fell recumbent and unresponsive and then started to flail. Experiment 2 performed the same test except the gas chamber held N2O prefilled at 25%, 50%, or 75% or CO2 prefilled at 7%, 14%, or 21%. The test was terminated more quickly at higher concentrations due to the piglets' responses. Time spent ataxic was greater in the middle concentration gradients. Flailing behavior tended to correlate with increasing concentrations of CO2 but not N2O. Experiment 3, using the minimal anesthesia model, showed that both 90 N and 90 C induced isoelectric EEG, in 71 and 59 s respectively, but not 60 N within 15 min. The EEG results together with the observed behavioral changes reflect differences in the animal's perceptive experience. The implications for animal welfare are that N2O is much less aversive than CO2, and 90% N2O can euthanize piglets.