PD CEN/TS 16817-2:2015

European foreword
Introduction
1 Scope
2 Normative references
3 Terms and definitions
4 Basic principle of the procedure
5 Sample matrices
6 Sampling procedure
7 Palynology
8 Molecular-biological analysis
9 Determination of the target parameters for
   GMO monitoring and representation of the results
10 Performance characteristics of the methods
11 Quality assurance and quality control
Annex A (normative) - Maize-specific requirements
Annex B (normative) - Rapeseed specific requirements
Annex C (informative) - Good beekeeping practice
Bibliography

Explains a procedure through which pollen - in particular pollen of genetically modified organisms (GMO) - can be sampled by means of bee colonies.

This Technical Specification describes a procedure through which pollen – in particular pollen of genetically modified organisms (GMO) – can be sampled by means of bee colonies.

Bee colonies, especially the foraging bees, actively roam an area and are therefore area related samplers. Pollen sampling depends on the collection activity of the bees and the availability of pollen sources within the spatial zone according to the bees\' preferences (supply of melliferous plants). A colony of bees normally forages over an area of up to 5 km radius (median 1,6 km, mean 2,2 km), in rare cases some bees may also forage in greater distances up to 10 km and more [26].

Foragers fix the gathered pollen on the outside of their hind legs (pollen loads, also known as pollen pellets). Inside the hive they place these pollen loads into comb cells close to the brood nest (bee bread). Furthermore, foragers gather nectar and honeydew. Nectar contains pollen which fell from the anthers of the blossom into the nectar drop, or pollen which was dispersed by the wind and sticks in the nectar of other blossoms or adheres to the sticky honeydew of plants. Nectar and honeydew are converted to honey and stored by the bees in the beehive.

Honey, pollen load and bee-bread may be used as sample matrices for the subsequent analysis of pollen as it is possible to concentrate sufficient amounts of pollen for microscopic and molecular biological diagnostics.

Microscopic analysis is used to identify the various pollen types and to quantify the exposure to the target pollen types in question. GMO exposure is analysed by molecular-biological methods: For analysis of pollen DNA quantitative PCR methods are used and described here in this Technical Specification. The analysis of GMO specific proteins and toxins in pollen is possible, too, using ELISA, but to this date the method has not been evaluated enough in pollen matrices for standardization in this Technical Specification.