5.3. in vitro larval tests

Aupinel et al. (2005) devised a standard in vitro test usable for any research topic on larvae (Crailsheim et al., 2013) and more specifically for brood risk assessment (Aupinel et al, 2007b). This test has already been ring-tested (Aupinel et al, 2009) with the participation of 7 laboratories originating from 6 countries that satisfied the 2 criteria of validity: control mortality lower than 15% at D6 and successful emergence of worker adults in at least the control group. This test, based on an individual rearing method permits one to control exactly the individual exposure with a high reproducibility. It provides quantitative oral toxicity data on honey bee brood. It is designed for in vitro treatments of AIs or formulated pesticides. Adopted in France by the CEB, it was validated at OECD and will be recommended in the near future as a guideline for acute exposure at D4 and lethal effect at D7. Chronic exposure and observations on pupae and adult stages will be referenced as guidance.