1) Mortality can be defined according to the following criteria:
- Larva: an immobile larva or a larva which does not
react to the contact of the paintbrush is noted as dead.
- Pupa: a non-emerged individual at D22 is noted as dead during pupal stage.
- Adult: an immobile adult which does not react to a tactile stimulation is noted as dead.
2) Mortality is checked at the following moments:
- Larva: At the feeding moment, dead larvae are
systematically removed for sanitary reasons. Specific mortality checks are made
according to the type of test. In the test where exposure is at D4 (acute
toxicity), a first mortality check is made at D4 in order to replace the dead
larvae before they have started consuming the diet containing the insecticide.
Then one should note the mortality at D5, D6 and D7. In the test with chronic
exposure, mortality is noted at D7.
- Pupa: Non emerged bees are counted at D22.
- Adult: Alive adult bees and dead adults which have left their cell and show a normal development are both counted at D22.
3) Sublethal effects such as development length, prepupa weight, wing malformation, adult survival, etc. can be noted. It is recommended to weigh prepupa without removing them from the rearing plastic cell. Adults can be kept in the emergence boxes with ad libitum food for behaviour observations or longevity assessment.