4.3.5.2.8. Bees’ exposure

The exposure of bees to AI following SSST is more difficult to control than that following spraying of non-systemic products. This is because blooming does not occur in the whole surface at the same time and because during the blooming period one cannot say if bees are only going to forage in the treated crop. Therefore, special manipulations need to be performed to ensure the level of exposure achieved by the colony as a whole. The control of the colony’s food intake is one parameter that can be achieved.

For this purpose, pollen pellets should be collected with pollen traps installed at the entrance of the colonies prior the blooming of the first flowers of the crop and every 2-3 days during the blooming period (see Human et al., 2013). Samples of at least 5 g  of pollen should be collected and kept in hermetic conditions, adequately labelled and immediately frozen. Samples are stored at least at -18ºC before analysis.

Pollen from the comb should be collected once before the beginning of the crop bloom and once a week following it. If the samples were taken by cutting a piece of comb, wax samples would be readily available. Otherwise, wax samples should be taken as well on the same days and immediately frozen. Samples are stored at least at -18ºC before analysis.

Foragers returning to their hive should be collected (see section 4.3.4.1.2.) at the entrance of the colony to undergo residue analysis of the content of their honey sac. Approximately 50 foragers should be collected prior to the blooming and every 2-3 days during the blooming period. Samples should be kept in hermetic conditions, adequately labelled and immediately frozen. Samples are stored at least at -18ºC before analysis.

Honey samples should be collected once before the blooming of the crop and once a week after.

Dead bees should be counted daily from the period starting before the bloom and 42 days after it. Dead bee traps (Human et al., 2013) will be cleaned every evening and samples of bees should be collected from the bee traps before sunrise. The collection period goes from just before the start of blooming and is conducted every 2 days during the blooming period. Samples should be kept in hermetic bags, appropriately labelled and immediately frozen (stored at least at -18ºC before analysis).

The quantity of sample per beekeeping matrix hereby proposed is indicative. It should be checked with the laboratory in charge of residue analyses prior to the beginning of the test.

Prepupae should be counted daily, in the same way as dead bees. Bee traps will be cleaned every evening and samples of bees should be collected from the bee traps before sunrise. They can be collected from the bee traps every 2 days and kept in hermetic bags, appropriately labelled and immediately frozen. Another option is the sampling of larvae directly from the comb once before the blooming of the crop and once a week after. Again samples should be kept in hermetic bags, appropriately labelled and frozen in case analyses should be delayed.

4.3.5.2.8.3. Reserves of the colonies at the beginning of the trial

The BEEBOOK