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- OECD 202: Daphnia sp., Acute Immobilisation Test
- OECD 211: Daphnia magna Reproduction Test
- OECD 235: Chironomus sp., Acute Immobilisation Test
- OECD 218/219: Sediment-Water Chironomid Toxicity Test Using Spiked Sediment/Spiked Water
- OECD 233: Sediment-Water Chironomid Life-Cycle Toxicity Test Using Spiked Water or Spiked Sediment
- OECD 225: Sediment-water Lumbriculus Toxicity Test Using Spiked Sediment
- OECD 242: Potamopyrgus antipodarum Reproduction Test
- OECD 243: Lymnaea stagnalis Reproduction Test
- Fish and other vertebrates
- OECD 203: Fish, Acute Toxicity Test
- OECD 215: Fish Juvenile Growth Study
- OECD 212: Fish, Short-term Toxicity Test on Embryo and Sac-fry Stages
- OECD 231: The Amphibian Metamorphosis Assay
- OECD 236: Fish Embryo Acute Toxicity Test
- OECD 210: Fish, Early-life Stage Toxicity Test
- OECD 229 Fish Short Term Reproduction Assay and OECD 230 21-day Fish Assay
- OECD 240 Medaka Extended One Generation Reproduction Test (MEOGRT)
- OECD 248: Xenopus Eleutheroembryonic Thyroid Assay
- OPPTS 850.1500: Fish Life Cycle Toxicity Test
- OÈCD 234 Fish sexual development test
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- OPPTS 830.6302, OPPTS 830.6303,and OPPTS 830.6304: Physical State, Colour and Odor at 20 °C and at 101.3 kPa
- EU A.1: Melting temperature/range
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- EU A.9: Flashpoint
- EU A.10: Flammability (solids)
- EU A.12: Flammability (contact with water)
- EU A.13: Pyrophoric properties of solids and liquids
- EU A.16: Relative self-ignition temperature for solids
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- OECD 114: Viscosity of Liquids
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- Non-target Arthropods
- Non-target arthropod testing with the parasitic wasp (Aphidius rhopalosiphi)
- Non-target arthropod testing with the lacewing (Chrysoperla carnea)
- Non-target arthropod testing with the ladybird beetle (Coccinella septempunctata)
- Non-target arthropod testing with the predatory bug (Orius laevigatus)
- Non-target arthropod testing with the predatory mite (Typhlodromus pyri)
- Non-target arthropod testing with the rove beetle (Aleochara bilineata)
- Non-target arthropod testing with the carabid beetle (Poecilus cupreus)
- Non-target arthropod testing with the wolf spider (Pardosa spec.)
- Soil Organisms
- Honey Bees and other Pollinators
- OECD 213/214: Honey bees, Acute Oral and Acute Contact Toxicity Test
- OECD 245: Honey Bee (Apis Mellifera L.), Chronic Oral Toxicity Test (10-Day Feeding)
- OECD 237: Honey Bee Larval Toxicity Test, Single Exposure
- OECD 239: Honey Bee Larval Toxicity Test
- EPPO 170: Honey Bee Field Study – do plant protection products effect honey bee colonies?
- Oomen et al. 1992: Honey Bee Brood Feeding Study
- OECD 75: Honey Bee Brood Test under Semi-field Conditions in Tunnels
- OECD 246/247 Acute Oral and Contact Toxicity to the Bumblebee, Bombus terrestris L.
- Solitary Bee Acute Contact Toxicity Study in the Laboratory (Osmia sp.) Solitary Bee Acute Oral Toxicity Study in the Laboratory (Osmia sp.) (protocols for ringtests with solitary bees recommended by the non-Apis working group)
- SANTE/11956/2016 rev.9 Residue trials for MRL setting in honey
- Non-target plants
- OECD 208: Terrestrial Plant Test - Seedling Emergence and Seedling Growth Test
- OECD 227: Terrestrial Plant Test - Vegetative Vigour Test
- OCSPP 850.4100: Seedling Emergence and Seedling Growth
- OCSPP 850.4150: Vegetative Vigor
- EPPO PP 1/207(2): Efficacy evaluation of plant protection products, Effects on succeeding crops
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EPPO 170: Honey Bee Field Study – do plant protection products effect honey bee colonies?
Honey bees (Apis mellifera L.) have a very important role in pollinating flowers and orchards. Their honey is used as a food since thousands of years. For these reasons international legislation claims for the protection of honey bees from any possible side-effect of agrochemical applications. A field study on honey bees under realistic conditions of agricultural practice (GAP) is indicated if laboratory studies and semi-field studies cannot sufficiently prove harmlessness of plant protection product applications. A field study allows to assess the impact of plant protection products on honey bee colonies under real agricultural conditions and thus is the final tool to show whether possible effects are acceptable.
Study Design
Test organisms
Healthy honey bee colonies, bred in normal beekeepers manner, of appropriate size should be produced to guarantee that the colonies are as uniform as possible. The quantity of food stores within the hive at the beginning of a study should be kept at a minimum to ensure larvae being fed by contaminated food.
Test Concentrations
Only formulated products are tested and should normally be applied at the highest field rate intended for the registration of the product in order to produce a worst-case exposure of the bees. A field test include at least a field treated with the plant protection product and an untreated control field (no reference item treatment is used).
Field Sites
The field sites should met the following criteria:
- At least two field sites (1 test item field and 1 control field)
- A size of at least 2500 m2 in case of Phacelia tanacetifolia or 1 ha in case of Brassica napus
- The total number of honey bee colonies should be in relation to the size of the fields
- The field sites should be seperated in a suitable distance (at least 2 to 3 km)
- Preferable, no main massive bee attractive crops should be in a radius of 2 km close to the surrounding of the field sites
Course of the test
At least four bee colonies per test group, bred in normal beekeepers manner, will be placed on the edge of the fields a few days prior to the application. The crop should be full flowering with bees actively foraging on the crop. On the day of application, the whole plot is sprayed according to GAP with calibrated field spraying equipment. The control plot remains untreated.
After 7 days of direct exposure of the colonies to the treated crop, the bee colonies are removed from the field sites to an area with no main attractive crops. The full testing period is up to 28 days following the application day.
Optional: Residue analysis
According to regulatory requirements residue studies may be required to determine residue levels in:
- pollen
- nectar
- flowers
- and/or foraging bees
In order to proove exposure, residue samplings can be incorporated into field studies with additional hives used exclusevily for residue purposes. Bee hives used for residue samplings are not biologically assessed (e.g. mortality, foraging activity, colony assessments, etc.). Details about the residue samplings are described in the recent study protocol (e.g. timing, amount, etc.).
Endpoints /Parameters
- Meteorological data
- Foraging activity: in marked areas of 1 m2 the number of bees are counted during e.g. 10-15s.
- Mortality: mortality in the hives are assessed by counting dead bees in dead bee traps. In addition, sheets (3 × 1.5 m) can be spread out infront of the bee hives in order to facilitate the collection and counting of dead bees infront of the bee hives.
- Behavioural abnormalities
- Condition of the colonies: the state of the adult bee population, the viability of the queen and the area of brood and food is assessed.
- Optional: Assessment of bee brood development (adaptations according to the OECD guideline no. 75). Please refere to “OECD 75: Honey Bee Brood Test under semi-field conditions”.
Guidelines and Literature
- OEPP/EPPO guideline No. 170 (4) (OEPP/EPPO, 2010)
- Optional: adaptations to OECD 75: “Guidance Document on the Honeybee (Apis Mellifera L.) Brood Test under Semi-Field Conditions"
- AG Bienenschutz (2011)