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Simple SummaryCarbon dioxide is commonly used for stunning animals prior to killing. It allows several animals to be killed at once, reduces the need for handling, and is a reliable method. However, research in laboratory rodents, poultry, and pigs has indicated that it causes considerable aversion at concentrations above ambient conditions. Currently, there are no available alternatives with desirable characteristics. This manuscript describes a list of research priorities to find and implement the use of alternative methods or agents to improve animal welfare.AbstractThe use of carbon dioxide (CO2) for stunning and killing animals is considered to compromise welfare due to air hunger, anxiety, fear, and pain. Despite decades of research, no alternatives have so far been found that provide a safe and reliable way to induce unconsciousness in groups of animals, and also cause less distress than CO2. Here, we revisit the current and historical literature to identify key research questions that may lead to the identification and implementation of more humane alternatives to induce unconsciousness in mice, rats, poultry, and pigs. In addition to the evaluation of novel methods and agents, we identify the need to standardise the terminology and behavioural assays within the field. We further reason that more accurate measurements of consciousness state are needed and serve as a central component in the assessment of suffering. Therefore, we propose a roadmap toward improving animal welfare during end-of-life procedures.

The minimum alveolar concentration of desflurane when combined with a continuous infusion of medetomidine at 3.5 microg/kg/hour was measured in seven ponies. Anaesthesia was induced with medetomidine (7 microg/kg intravenously) followed by ketamine (2 mg/kg intravenously) and maintained with desflurane in oxygen. The infusion of medetomidine was started 20 minutes after the induction of anaesthesia. The electrical test stimulus was applied at the coronary band (50 V, 10 ms bursts at 5 Hz for one minute), and heart rates and rhythms, arterial blood pressures, and arterial blood gas tensions were measured at intervals, just before the application of the stimulus. The mean (sd) minimum alveolar concentration of desflurane was 5.3 (1.04) per cent (range 3.2 to 6.4 per cent), 28 per cent less than the previously published value for desflurane alone after the induction of anaesthesia with xylazine and ketamine. The cardiopulmonary parameters remained stable throughout the period of anaesthesia. The mean (sd) time taken by the ponies to stand after the administration of desflurane ceased was 16.5 (6.17) (range 5.8 to 26) minutes, and the quality of recovery was good or excellent. However, one pony died shortly after standing; a postmortem examination revealed that it had chronic left atrial dilatation.

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This study evaluated the breathing pattern and distribution of ventilation in horses prior to and following recovery from general anaesthesia using electrical impedance tomography (EIT). Six horses were anaesthetised for 6 hours in dorsal recumbency. Arterial blood gas and EIT measurements were performed 24 hours before (baseline) and 1, 2, 3, 4, 5 and 6 hours after horses stood following anaesthesia. At each time point 4 representative spontaneous breaths were analysed. The percentage of the total breath length during which impedance remained greater than 50% of the maximum inspiratory impedance change (breath holding), the fraction of total tidal ventilation within each of four stacked regions of interest (ROI) (distribution of ventilation) and the filling time and inflation period of seven ROI evenly distributed over the dorso-ventral height of the lungs were calculated. Mixed effects multi-linear regression and linear regression were used and significance was set at p<0.05. All horses demonstrated inspiratory breath holding until 5 hours after standing. No change from baseline was seen for the distribution of ventilation during inspiration. Filling time and inflation period were more rapid and shorter in ventral and slower and longer in most dorsal ROI compared to baseline, respectively. In a mixed effects multi-linear regression, breath holding was significantly correlated with PaCO2 in both the univariate and multivariate regression. Following recovery from anaesthesia, horses showed inspiratory breath holding during which gas redistributed from ventral into dorsal regions of the lungs. This suggests auto-recruitment of lung tissue which would have been dependent and likely atelectic during anaesthesia.