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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
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- 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 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)
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- 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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OECD 202: Daphnia sp., Acute Immobilisation Test
Water flees are important grazers feeding on phytoplancton. Therefore they are part of risk assessments for several kinds of substances like plant protection products, industrial chemicals, biocides and medicinal products for human and veterinary use. Additionally, they are easy to rear and sensitive to water pollution.
Study Design
Test organism
The water flea Daphnia magna belongs to the Crustacea, subclass Phyllopoda. Young daphnids not older than 24 hours are tested from a synchronous laboratory culture.
Course of the acute immobilisation test
The acute toxicity test determines immobility during an exposure period of 48 hours. Immobilisation and abnormal behaviour is recorded at 24 and 48 hours and compared with control values. The test item is mixed into reconstituted water. 20 Daphnia are divided in four replicates per control and test item concentration. Most usually a dose-response study with a geometric series of five test item concentrations is tested. For the test, a static semi-static or flow-through design can be used.
Exposure takes place in a controlled environment room with defined temperature and light regime. Dissolved oxygen, pH and water temperature are measured regularly. The test item concentrations
are checked by analytical dose verification.
Alternative species testing
In order to determine a species sensitivity distribution (SSD), additional species can be tested in adapted test designs on request e.g. Chironomus riparius, Ceriodaphnia dubia, Tetrahymena thermophile, Brachionus calyciflorus or Thamnocephalus platyurus.
Endpoints
LOEC, NOEC, EC50 (EC20, EC10) for immobility of the daphnids are determined.
Guidelines and Literature
- OECD 202: Daphnia sp., acute immobilsation test
- Council Regulation (EC) No 440/2008, C.2. Daphnia sp., Acute Immobilisation Test
- EPA Ecological Effects Test Guideline OPPTS 850.1010 Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids