The foliage dwelling insect Coccinella septempunctata is recommended as additional specie where the in-field or off-field risk assessment for the 2 standard species fails.
Ladybird beetle larvae are exposed to different concentrations of the test substance, applied on glass plates (laboratory test) or plant leaves (extended laboratory), by calibrated spray equipemnt, until the hatching of the adults in order to evaluate the effects in terms of mortality over the exposure period and the reproduction performance of the adults in terms of egg deposition and larval hatching rate over the following 2 weeks.

TEST SYSTEM
Larvae of Coccinella septempunctata

STUDY DESIGN
Range finding is carried out with at least 3 application rates of the test substance and 1 untreated control (at least 15 replicates/treatment or control)  with 1 larva/replicate.
Limit test includes 1 application rate of the test substance, 1 untreated control and 1 toxic reference treatment (40 replicates/treatment or control) with 1 larva/replicate.
Rate-response test includes at least 5 application rates of the test substance, 1 untreated control and 1 toxic reference treatment (30 replicates/treatment or control) with 1 larva/replicate.

In order to evaluate the duration of the effects of the test substance and the recovery capacity of the insects it is possible to carry out an extended laboratory test with aged residues of the test substance.
The plants are treated in semi-field conditions and the leaves of the treated plants, after an adequate aging time, are moved to laboratory to perform the standard test as previously described.

ENDPOINTS
Pre-imaginal mortality: NOER/LR₅₀ over the exposure period.
Reproductive performance: NOER/ER₅₀ for eggs production and for eggs fertility over 2 weeks  following the exposure.
Study includes GLP management and reporting.

REFERENCES AND GUIDELINES
IOBC/WPRS Guidelines (Mead-Briggs et al., 2000).
ESCORT I Guidance Document (Barret K.L. et al., eds. 1994).
ESCORT II Guidance Document (Candolfi et al., eds. 2001).