02427nas a2200241   4500000000100000000000100001008004100002260007000043653005100113653001700164653001500181653001200196653001300208100001100221700001300232700001400245700002100259700001300280245011000293490000800403520176000411022001402171       2018                            d  c06/2018bSchlütersche Verlagsgesellschaft mbH & Co. KGaHannover10aProtein misfolding cyclic amplification (PMCA)10a263K scrapie10astereotypy10ahousing10ahandling1 aL Paul1 aP Kirsch1 aA Thomzig1 aC Thöne-Reineke1 aM Beekes00aPractical approaches for refinement and reduction of animal experiments with bank voles in prion research0 v1313 aThe bank vole (Myodes glareolus) is by far not as common a laboratory rodent  as mice or rats. Recently, however, bank voles have increasingly emerged as a  powerful infection model for prion diseases such as human Creutzfeldt-Jakob  disease (CJD) or chronic wasting disease (CWD) of cervids. The susceptibility of  bank voles to infections with a variety of prion isolates from different host species  will substantially facilitate research into prion diseases and their causative agents.  To pro-actively promote a reduction of prion bioassays in bank voles against this  background, we examined protein misfolding cyclic amplification (PMCA) using  brain homogenate from normal bank voles as reaction substrate for its suitability  as a possible in vitro alternative. PMCAs using bank vole brain homogenate as  substrate (bvPMCAs) are known to be highly vulnerable to false-positive results.  Such false-positive readouts predominantly result from inadvertent crosscontamination  and unspecific seeding with minute amounts of prion agents  such as 263K scrapie which are often present in prion research laboratories. For  tackling this problem that potentially impedes a broader application of bvPMCA,  we describe bvPMCA conditions that were ultra-sensitive to 263K scrapie prions  and yet highly robust against inadvertent cross-contamination. With respect to  refinement, we implemented refined housing conditions that prevented the occurrence of stereotypies in bank voles, and procedures which allowed a virtually  stress-free handling when subjecting these animals to laboratory procedures such as weighing or inhalation of anaesthetics. These refinements can improve  the welfare of bank voles in prion laboratories and other fields of research.  a0005-9366