- Aquatic Ecotoxicology
- 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: Lymnea stagnalis Reproduction Test
- 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 241: Larval Amphibian Growth and Development Assay
- OPPTS 850.1500: Fish Life Cycle Toxicity Test
- OÈCD 234 Fish sexual development test
- Storage Stability Studies
- 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
- EU A.2: Boiling temperature
- EU A.3: Relative density (liquids and solids)
- EU A.4: Vapour pressure
- EU A.5: Surface tension
- 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
- EU A.17: Oxidising properties
- OECD 114: Viscosity of Liquids
- Environmental Fate
- Terrestrial Ecotoxicology
- 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.)
- 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
- 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
- Ecological Modelling
- Quality Assurance
- Testing of Potential Endocrine Disruptors
- Aquatic Ecotoxicology
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OECD 207/222 Earthworm Acute Toxicity and Reproduction Test
Earthworms play an important role in the soil biocenosis since they enhance soil characteristics such as the soil function of litter breakdown, soil fertility and soil microstructure. Therefore, earthworms are used as a standard test species to investigate the impact of a substance that is applied on soil or when a contamination of soil is possible. Usually, is in a first step the acute toxicity tested. Persistent substances or substances with several applications per season cause a longer exposure for the earthworm community. In these cases the performance of a study on sublethal effects is required.
Eisenia fetida (Savigny 1826) belongs to the Annelida, Oligochaeta. The test organisms originate from an internal synchronous laboratory breeding culture. Adult hermaphrodites are used for testing when they are between two and twelve months old
Course of the Test
Earthworm tests are performed with an artificial soil (according to OECD requirements) consisting of quartz sand, sphagnum peat, kaolin clay and calcium carbonate. Testing is performed in a climatic chamber with defined temperature, light regime and intensity.
OECD 207: Earthworm, Acute Toxicity Tests
On the basis of the results of a non-GLP pre-test, the applied concentrations for the main test are selected. The test item is mixed into the artificial soil.
Usually 5 concentrations in a geometric row with 4 replicates is performed. In total 40 earthworms are exposed to the test item per treatment group i.e. ten individuals per replicate. Each test comprises a negative control. A test with a toxic reference substance is performed once a year to proofs the sensitivity of the test system. After 7 and 14 days of exposure the mortality of earthworms is assessed.
As sublethal parameters the mean body weight and behavioural abnormalities are recorded.
OECD 222: Earthworm Reproduction Test
Usually 5 test substance concentrations (8 concentrations for ECx-design) with four replicates each are tested (control 8 replicates). The application can be performed by mixing the substance into the artificial soil or by spraying onto the soil surface. For plant protection products the main focus of this study is to show that the no observed effect concentration (NOEC) of the product is less than the 5 fold predicted environmental concentration (PEC), which is set as the acceptability threshold. Otherwise, an earthworm field study is the option to prove environmental safety.
10 adult worms (not older than 1 year) are exposed for 4 weeks. Mortality and weight changes are assessed after this time. The offspring is exposed another 4 weeks until a total exposure period of 8 weeks is reached.
Derivation of a concentration-response curve for mortality. The LC50 and if possible a NOEC are determined statistically.
As the focus of this study lies on reproduction, the tested concentration range mostly results in slight or no mortality. Reproductive success of the worms. NOEC and/or ECx are determined statistically. Additionally other sublethal effects like body weight changes and feeding activity are recorded.
Guidelines and Literature
- OECD-Guideline for the testing of chemicals No. 207 “Earthworm, Acute Toxicity Test” (adopted April 4, 1984).
- ISO-Guideline 11268-1:1993 “Soil quality – Effects of pollutants on earthworms (Eisenia fetida) - Part 1: Determination of acute toxicity using artificial soil substrate”.
- OECD, Guideline for the testing of chemicals Nr. 222 "Earthworm, Reproduction Test" (adopted April 13, 2004).
- ISO-Guideline 11268-2, “Soil quality - Effects of pollutants on earthworm (Eisenia fetida) - Part 2: „Determination of effects on reproduction“ (1998).