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              <p>Magnetic Resonance Imaging (MRI) is a non-invasive technique that requires the participant to be completely motionless. To date, MRI in awake and unrestrained animals has only been achieved with humans and dogs (Andics et al., 2014). For other species, alternative techniques such as anesthesia (Barrière et al., 2019), restraint (Stefanacci et al., 1998) and/or sedation (Behroozi et al., 2018) have been necessary. MRI studies with sheep have only been conducted under general anesthesia. This ensures the absence of movement and allows relatively long MRI experiments. However, it can also be detrimental to health, disrupt neurovascular coupling (Aksenov et al., 2015) and does not permit the study of higher-level cognition. Here we present a proof of concept that positive reinforcement training in sheep is sufficient to be able to acquire MRI of their brain.We developed a step-by-step training protocol based on positive reinforcement (food reward, social reward) and human – animal cooperation with 10 lambs from birth. This training method involved two phases: 1) training in the sheepfold using a mock MRI and 2) training in the real MRI environment. Each phase consisted of successive steps during which sheep learnt a behavior necessary for them to be able to participate in an MRI experiment. To successfully complete this training protocol, sheep must remain motionless lying on the MRI table for the minimum time required for an acquisition. At present all 10 sheep actively participate in training and place their head inside the MRI coil for a minimum duration of ~1 min 30 sec. Six have done so long enough to acquire both T1-weighted structural (~2 min 30 sec, Figure 1) and EPI functional images (~4 min 30 sec).These promising early results clearly show that MRI data can be acquired with awake and unrestrained sheep. The innovative training protocol opens the door to the possibility of conducting animal welfare friendly functional MRI studies with sheep. We are currently preparing a first awake functional MRI study investigating their auditory system.</p>
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