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Pflanzenschutzmittelrückstände im gehöselten Pollen der Honigbiene (Apis mellifera L.) : Auswirkungen einer feldrealistischen Pflanzenschutzmittelmischung auf Stockbienen und den Larvenfuttersaft
(2017) Böhme, Franziska; Zebitz, Claus P. W.
Pesticides are used worldwide and contaminate air, surfaces, soils and the aquifer. Non-target-organisms and non-target-plants may get into contact with pesticides di-rectly via drift or indirectly via run-off, leaching or sowing dust. Due to pollination services and bee products, the honeybee (Apis mellifera L.) is a non-target-organism of major interest for humans. On their flights around the beehive they collect water, pol-len, nectar, honeydew and tree resin. The proteins originating from the pollen are im-portant for nutrition and development of larvae and adults. Pollen is stored and fer-mented inside the hive as beebread and is made of hundreds of pollen loads of differ-ent plants collected over a longer period. Pesticide residue analyses of beebread is a common tool to estimate the contact of honeybees to pesticides in the field. However, such beebread analyses cover a larger time frame and a mixture with uncontaminated pollen will dilute the maximum residue levels of certain plant pollen. Therefore, pesti-cide analysis of bee bread is only an approximate approach to estimate the real pesti-cide exposition. Thus, pollen pellets were collected daily at three distinct sites with differences in agri-cultural intensity in Baden-Württemberg from 2012 - 2016 during the agronomic active season (spring/summer). We wanted to give detailed information on the daily contact to pesticides as well as changing pesticide frequencies and combinations throughout the season. 281 pollen pellet samples, each representing a single day, were analyzed for 282 active ingredients currently used in agricultural practice (publication 1). Huge qualitative and quantitative differences in the pesticide load between the sites were discovered. The meadow site near Göppingen was the least contaminated. In five ob-servation years only 24 different substances were found in 56 % of the samples with concentrations up to 300 µg/kg. The more intensive site in Ertingen is characterized by grains and maize for biogas plants. Only 13 % of the samples were uncontaminated, in the remaining samples 37 substances with maximal concentrations up to 1,500 µg/kg were detected. The site with the highest occurrence of crop protection was close to Heilbronn. Permanent crops such as wine and orchards shape the landscape. The high-est detected concentration was 7,178 µg/kg. All samples were contaminated with up to 58 different substances. During the five years of observation 73 different pesticides were found. Due to admis-sion regulations, there was a high likelihood to find 84 % of these substances in pollen. Twelve substances were found that are either not registered as plant protection prod-ucts or are not supposed to get in contact with bees. This indicates a need for further improvement of seed treatments and increasing awareness of flowering shrubs, field margins and pesticide drift. Concluding from the majority of concentrations and pesti-cides found, we assume no misuse of pesticides by the farmers at our three sites in the observation period, which would lead to direct intoxication. Considering LD50 values, the here detected concentrations are sub-lethal for honeybees. However, at any tested site and in most of the samples a mixture of different pesticides was found. Yet, it is not known, whether there are effects caused by a combination of different pesticides in sub-lethal concentrations when consumed chronically by honeybees. Therefore, we conducted a field experiment with free-flying honeybee colonies (publi-cation 2). Mini-hives containing about 2,500 bees and sister queens were established at the Apicultural State Institute. Queens were confined to an empty frame to receive lar-vae of known age. These bees were intended to feed on pesticides chronically in two crucial life stages. After larvae hatched from the eggs and after adults hatched from the cells they were fed a pollen-honey diet contaminated with a cocktail of twelve dif-ferent active ingredients in field-realistic concentrations. In colonies treated with a pes-ticide mixture, larval weight was higher and acini diameters of the hypopharyngeal glands of nurse bees were smaller than in the untreated control. However, brood termi-nation and adult lifespan did not differ between both groups. Despite feeding a pesti-cide cocktail chronically starting on the first day of larval being, no obvious negative side-effects in worker bees were detected. It raises the question, if nurse bees, which feed on the contaminated pollen-honey diet, produce larval food and feed larvae, serve as a filter system so that larvae would not come into contact with the pesticides. To determine the fate of pesticides originating from the pollen source, we started a queen rearing (publication 3). Frames with 24 h old larvae were hang into queenless free flying mini-hives. At the same time, the colo-nies were fed a pollen-honey diet containing a cocktail of 13 commonly used pesti-cides in high concentrations. The royal jelly (RJ) fed to the larvae by nurse bees was harvested from the queen cells and subjected to a multi-pesticide residue analysis. Sev-en substances were rediscovered in traces (76.5% of all detections were below 1 μg/kg). However, worker larvae older than three days receive a modified jelly, containing pol-len coloring the food yellowish. That is why we were wondering if contaminated pol-len might have a different effect on the food of worker larvae. Queens of free-flying mini hives were caged to receive larvae of known age. The colonies received a pollen-honey diet, contaminated with high concentrations of a pesticide mixture (publication 4, submitted). Worker jelly (WJ) was harvested on four successive days from larval age three to six and subjected to a multi-pesticide residue analysis. Pesticide concentrations increased with larval age and ranged between 2.9 and 871.0 µg/kg for the different substances and age groups. As the increase of substances in the WJ positively corre-lates with the amount of pollen grains counted in the larval food, we were able to show a direct relationship between the administered pollen in the food and the pesticide concentrations. Considering the maximum food uptake rates of a worker larvae, even the highest con-centrations found, would lead solely to sub-lethal amounts. Even for queens, who con-sume RJ not only as larvae but during their whole life would consume only sub-lethal pesticide concentrations. Especially considering the not-field realistic concentrations we chose for our experiments. Probably, the sub-lethal effects found in our first exper-iment are due to the sub-lethal concentrations worker larvae have taken up chronically during their development. Even though we did not detect acute intoxication symptoms and the concentrations in the brood food are sub-lethal, we cannot infer whether there are impairments of fitness or brood success of honeybee colonies in the long term. However, as honeybee colonies are considered as superorganisms, they are able to tol-erate stressors or the loss of individuals. Therefore, the detection of sub-lethal effects on colony-level in the field is difficult. Yet, a vast problem arises with solitary living insects, for example wild bee species, which are more prone to stressors such as pesti-cides. Solitary insects have more restricted flight and collecting areas, get into contact with pesticides in pollen directly as larvae and have almost no buffer capacities.