“…Tier-1 assays have been used to evaluate the risks to NTOs of various Bt Cry proteins, lectins and some protease inhibitors. The test NTOs in these assays have included the honey bee (Aphis mellifera L.) (Babendreier et al, 2005;Hendriksma et al, 2011Hendriksma et al, , 2012, a solitary bee (Osmia bicornis L.) (Konrad et al, 2008), a bumble bee (Bombus terrestris L.) , ground beetles (e.g., Poecilus cupreus L.) (Duan et al, 2005;Stacey et al, 2006), ladybird beetles (e.g., Coleomegilla maculata De Geer, Adalia bipunctata L. and Coccinella septempunctata L.) (Li et al, 2011b;Álvarez-Alfageme et al, 2011Álvarez-Alfageme et al, , 2012, lacewings (e.g., Chrysoperla carnea Stephen and Chrysoperla sinica Tjeder) (Romeis et al, 2004;Lawo & Romeis, 2008;Li et al, 2008;Wang et al, 2012), the insidious flower bug (Orius insidiosus Say) (Duan et al, 2008a;Tan et al, 2011) and others. In the following sections, we discuss several key points that should be considered in the development of a sound Tier-1 assay.…”