Bee Safety of Neonicotinoids - Evidence from Studies Conducted Under Realistic Field Conditions

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Bee Safety of Neonicotinoids - Evidence from Studies Conducted Under Realistic Field Conditions Bayer CropScience, Environmental Safety - Ecotoxicology 7 Nov 2016 / Janine Doering

Introduction Neonicotinoid insecticides are used as seed treatment in various crops. Among these there are bee-attractive crops like oilseed rape, sunflower, and maize After neonicotinoid seed treatment, traces of systemic residues can be found in nectar and pollen of treated crops This has led to concerns about their safety to bees and other pollinators Although monitoring data do not suggest a systematic correlation between the exposure of bees to neonicotinoids and bee health issues, the use of neonicotinoids in bee-attractive crops has been restricted in the EU in 2013 In order to complement existing field data, a large scale field study with neonicotinoid-treated oilseed rape (treatment with Elado - 10 g clothianidin & 2 g β-cyfluthrin/kg seed) was conducted in Northern Germany Page 2

Setup and Purpose of the Study Conceptual approach of the monitoring study in Mecklenburg Vorpommern (Germany) Monitoring potential adverse effects at landscape level Comparable landscapes for control und treatment Spatial proximity of both landscapes: close enough to ensure comparable climatic and topographical conditions distant enough to avoid honeybees to cross-forage Agricultural landscapes with significant oilseed rape cultivation and low abundance of other flowering crops or wild plants distracting bees from the target crop Common oilseed rape cultivation practice with pre-agreed and defined crop protection measures Large-scale farm operations required to ensure regulated crop protection regimen Study design Monitoring of the clothianidin residue levels in nectar and pollen during bloom Monitoring potential adverse effects on pollinators which differ in life history traits Page 3

Similarity of Control and Treatment Sites Attractiveness for Bee Pollinators Two sites of approx. 65 km 2 each were selected according to field cropping area, statistic and land cover data Study area Control (Reference) 16% Oilseed rape Treatment 21% alternative forage areas without forrage sources Site C Site T Page 4

exposure time Similarity of Control & Treatment Sites Crop phenology Control fields received 1 2 additional pyrethroid applications to ensure comparable oilseed rape emergence Homogenous flowering period across control (reference) and treatment and over time Page 5

Similarity of Control & Treatment Sites Crop phenology Page 6

Characterization of Exposure Residue levels of Clothianidin in OSR nectar & pollen at bloom 1 tunnel = 10 m x 5 m 39 x 50 m² = 0.2 ha [18 fields, Total oilseed rape area 792 ha] 1.06 No residue detects in majority of oilseed rape pollen or nectar in control landscape (n = 34) Trace residue levels in few of control pollen samples (tunnel) & nectar samples (field) Residue levels in treatment landscape (n = 46/48 + 48) comparable to tunnel samples (n = 39) Page 7

Investigation of Potential Adverse Effects Three bee pollinator species with different life history traits Honeybee (Apis mellifera) Life history traits: o complex social community with task share o Comb cells of wax o Hives with 50.000+ workers o Thermoregulation o Full colony overwintering o Winter food stores Bumblebee (Bombus terrestris) Life history traits: o o o o o o Less complex social structure and task share Comb cells of wax Hives with 500+ workers Thermoregulation Only queens overwinter Limited food storage Red Mason bee (Osmia bicornis) Life history traits: o o o Solitary bee Nests in tube-shaped structures e.g. in deadwood Overwinter as pupa (cocoons) Page 8

Monitoring Results Honey Bees (Apis mellifera) Eight bee colonies per monitoring location (=1 replicate) Six replicates per site (treatment and control) Distribution of replicates per site: 3 replicates adjacent to oilseed rape fields, 3 replicates in 400 m distance. Monitoring location adjacent to field Post-exposure phase: west-central Germany Page 9

Monitoring Results Honey Bees (Apis mellifera) (1) Adult honey bees Typical seasonal colony development pattern No statistically significant effect of treatment or distance to fields Page 10

Monitoring Results Honey Bees (Apis mellifera) (2) Bee brood Typical seasonal growth pattern No statistically significant effect of treatment or distance to fields Page 11

Monitoring Results Honey Bees (Apis mellifera) (3) Honey yield (Spring) No statistically significant effect of treatment Statistically significant effect of distance to oilseed rape fields Page 12

Characterization of Exposure Sampling for residues Honey Bees (Apis mellifera) Tunnels: One Mini-Plus hive (30 x 30 x 30 cm, 6 combs, approx. 2500 bees) per tunnel, tunnels set-up inside study fields, sampling period 4 weeks, min. one pollen sample per tunnel (pollen traps, 100 mg / tunnel collected from morning to noon), nectar from 200 forager bees (content of honey stomachs), sample size 200 mg Free-flying: Eight Zander hives (50 x 43.5 x 23 cm, 10 combs, brood chamber plus 1-2 honey supers) at 12 locations = from 96 colonies, each colony sampled twice for pollen (pollen traps, 300 mg over one day), nectar from 200 forager bees (content of honey stomachs), sample size 200 mg 1.06 Page 13

Conclusions Honey Bees (Apis mellifera) Honey bees intensively foraged in treated fields (73-83% oilseed rape pollen found in honey) Residues of clothianidin in tunnels: average: 1.7 µg/kg in pollen (max. 3.5 µg/kg), average: 1.3 µg/kg in nectar (max 3.6 µg/kg), metabolites TZMU & TZNG < LOD Residue of clothianidin from field sites: average: 0.5 & 0.97 µg/kg in pollen (max. 2.7 µg/kg), average: 0.68 & 0.77 µg/kg in nectar (max. 1.6 µg/kg), average: 1.35 µg/kg in honey (max. 2.1 µg/kg in honey), metabolites TZMU & TZNG < LOD or LOQ Page 14

Conclusions Honey Bees (Apis mellifera) There were no treatment-related adverse effects observed on: colony strength and development (adults, brood) honey yield pollen composition infestation with parasites and diseases Page 15

Monitoring Results Bumble bees (Bombus terrestris) Three Tripols (= 3 bumblebee colonies) per monitoring location (= 1 replicate) Six replicates per site (treatment and control) Distribution of replicates per site: 3 replicates adjacent to oilseed rape fields, 3 replicates in 400 m distance. Monitoring location adjacent to field Collection of bumblebee workers for pollen sampling Page 16 Bayer 4:3 Template 2010 March 2016

Monitoring Results Bumble bees (Bombus terrestris) (1) Hive weight development No statistically significant effect of treatment or distance to oilseed rape fields Page 17

Monitoring Results Bumble bees (Bombus terrestris) (2) Number of worker bees per colony No statistically significant impact of treatment or distance to oilseed rape fields Page 18 Bayer 4:3 Template 2010 March 2016

Monitoring Results Bumble bees (Bombus terrestris) (3) Number of queens (including queen brood cells) No statistically significant effect of treatment or distance to oilseed rape fields Page 19

Monitoring Results Bumble bees (Bombus terrestris) (4) Residue detects No residues in control pollen samples Residue levels in treatment landscape (bumble bee) comparable to tunnels (honeybees) Page 20

Characterization of Exposure Sampling for residues Bumble bees (Bombus terrestris) One hive per study location used to sample 11-21 returning foragers, stored on dry ice, pollen loads were detached, sample size min 230 mg 1.06 Page 21

Conclusions Bumble bees (Bombus terrestris) Bumble bees intensively foraged in treated fields Besides oilseed rape pollen, pollen from willow, chestnuts and wild berries were identified Clothianidin residues in pollen: average: 0.88 µg/kg (max. 1.3 µg/kg) metabolites TZMU & TZNG < LOD There were no treatment-related adverse effects observed on: Colony development (workers, drones, queens) Seasonal growth pattern (turning point to queen production) Page 22

Monitoring Results Red Mason Bee (Osmia bicornis) Six Replicates per Control and Treatment with 3 nesting units adjacent to the oilseed rape field and 3 nesting units in 100 m distance 750 cocoons per nesting unit = 1500 cocoons per replicate released (sex ratio 6:5 m:f) Page 23

Monitoring Results Red Mason Bee (Osmia bicornis) Reproduction rate = Number of cocoons per nesting female (2014): Control: 7.47 Treatment: 8.61 1 Remark: Control replicate OCC not taken into account for reproduction analysis. Breeding activity was low due to incorrect positioning of nesting units (shadow location) that caused site-related poor performance Emergence rate spring 2015 Control : 91.0% Treatment: 92.2% No statistically significant difference to the control on reproductive performance nor on cocoons emergence of offspring Page 24

Characterization of Exposure Sampling for residues Red Mason Bees (Osmia bicornis) One sampling per study location, Nest blocks were opened to collect 10 subsamples from rear end of active provisioned nesting cell, Pollen retrieved with micro spoon, Combined sample amounted to min 200 mg 1.06 Page 25

Conclusions Red mason bees (Osmia bicornis) Red mason bees collected up to 32% of their pollen from oilseed rape fields (average 14%) Besides rape pollen, they collected pollen of roses and buttercups Clothianidin residues in pollen: average 0.88 µg/kg (max. 1.7 µg/kg) metabolites TZMU & TZNG < LOD There were no treatment-related adverse effects observed on: Reproductive performance (nesting activity, offspring production) Hatching success Page 26

Summary of Key Results Bee pollinators in the treatment site were exposed to low and representative levels of systemic Clothianidin residues Bee pollinators used oilseed rape pollen to different extent (honeybees > bumblebees > solitary bees) Honeybee (Apis mellifera) o o o No effects on colony development (adults & brood) No effects on honey yield and pollen composition No effects on infestation by parasites and diseases Bumblebee (Bombus terrestris) o o No effects on colony development (no. of workers, drones, queens) No effects on seasonal growth pattern (switch point to queen production) Red Mason bee (Osmia bicornis) o No effect on reproductive performance (nesting activity, offspring production, hatching success) Page 27

Overall Conclusions The chosen landscapes control and treatment were sufficiently similar in terms of topography, climate, soil types and crop phenology to deliver a robust conclusion on whether or not Clothianidin poses an unacceptable risk to populations of insect pollinators Residue levels in pollen and nectar of oilseed rape in the treatment landscape were in the range of residue levels found in previous studies, i.e. highly representative exposure situation There were no treatment-related short- or long-term adverse effects recorded for the Clothianidin seed treatment to three bee species with different life history traits Page 28

Six publications available (open access) Page 29

Thank you!

Further Large-Scale Field Studies Examples for other large-scale field studies under realistic field conditions: Studies in canola in Canada (2005 and 2012). Evaluation of colony health over one season and after overwintering Multi-site field study in maize in France. Locations in four different regions, study conduction over three consecutive years (2008-2010) Field study in Southwestern Germany (2008). Evaluation of colony health over one season and after overwintering. In none of these studies, adverse effects of neonicotinoid seed treatment products to honey bee colonies have been observed. Since field studies provide a direct insight into the interaction of pesticides with bee colonies, they provide highest-tier evidence for the ecotoxicological risk assessment Page 31