Zolazepam hydrochloride

Asian black bears were captured from May to June 2018 using barrel traps baited with honey. A total of five traps were set within 5 km of each other, and the trapped bears were immobilized with tiletamine hydrochloride and zolazepam hydrochloride (Virbac, Carros, France, 8 mg/1kg of body weight). Basic body measurements were taken, and premolar extraction was performed for age determination. Bears over 40 kg were equipped with microchips for individual identification and GPS/camera collars (GPS Vertex Lite Collars; Vectronic Aerospace GmbH, Berlin, Germany). The collars (1.2 kg) weight less than 3% of body mass of bears. Bear capture and handling were approved by the Institutional Animal Care and Use Committee of Tokyo University of Agriculture and Technology. All experimental procedures used in the bear research followed the Guidelines Concerning Animal Experimentation of the Tokyo University of Agriculture and Technology and the Mammal Society of Japan (Mammal Society of Japan 2020).

For the experiment permission was obtained from the medical ethical committee of the HongHui hospital, Xi’an, China. Ablations were performed in tibia of male sheep (Small-tail Han) aged 1–1.5 years with aimed mass of 50 kg (in vivo experiment: range 45–50 kg). Three settings were examined for both a one week- and a six weeks follow-up group: 60W 10min, 60W – 20 min and 80W – 10 min. For each group four sheep were included, leading to a total of 24 sheep. As controls the contralateral tibias were used. Sheep were randomly divided in the one- or six week group. Before starting the procedure (Fig 1A), the sheep were sedated with an intramuscular injection of 5 ml Shutai (1: 1 combination of Tiletamine 50mg/ml & Zolazepam hydrochloride 50mg/ml) (Virbac, France) and subsequently intubated (Fig 1B. During the complete procedure inhaled isoflurane (1–5%) was used for general anesthesia and a maintenance dose up to 2.5ml Shutai was given in case deemed necessary. For post-procedural analgesia meloxicam injections were used. After the follow-up period sheep were euthanized by an experienced veterinarian (bleed out under intramuscular sedation). Sheep with a (deep) infection or fracture were excluded from mechanical analysis to prevent finding differences in strength based on other factors than the mechanical effect of MWA on bone.