Table 1 Internationally accepted invertebrate toxicity test guidelines. OECD CF level refers to the OECD Conceptual Framework [58]
| Invertebrate group | Test guideline | OECD CF level | Endpoints | Comment |
|---|---|---|---|---|
| Annelida | OECD TG 220: Enchytraeid Reproduction Test | 4 |
Mortality No. juveniles / adult | |
| OECD TG 222: Earthworm Reproduction Test (Eisenia fetida; Eisenia andrei) | 4 |
Mortality Growth (weight) No. juveniles/adult | ||
| OECD TG 225: Sediment–Water Lumbriculus Toxicity Test Using Spiked Sediment | 4 |
Mortality Biomass Reproduction (total No. or increase in No.) | ||
| Chelicerata | OECD TG 226: Predatory mite (Hypoaspis (Geolaelaps) aculeifer) reproduction test in soil | 4 |
Mortality Reproduction (total No. of juveniles) | |
| Collembola | OECD TG 232: Collembolan Reproduction Test in Soil | 4 |
Mortality Reproduction (total No. of juveniles) | |
| Crustacea |
Short-Term Juvenile Hormone Activity Screening Assay using Daphnia magna (SJHASA) (draft OECD TG) | 3 | Production of male neonates | Under development |
| OECD TG 211/OCSPP 850.1300: Daphnia magna Reproduction Test (with potential male induction assessed) | 4 |
Mortality No. live offspring/parent Time to first brood Parental growth (length and body weight; optional) Presence of ephippia or male neonates | Male production is known in response to juvenile hormone mimics. However, males are also produced under changing environmental and stressful conditions | |
| OPPTS 850.1350: Mysid Chronic Toxicity Test | Not listed |
Mortality Time to appearance of secondary sexual characters Time to first brood Growth (length and weight) Reproduction (No. young/female) | ||
| Mysid Life Cycle Toxicity Test (when OPPTS 850.1350 is finalised as an OCSPP guideline) | 5 |
Mortality Time to maturity Time to 1st & 2nd brood release Inter-brood duration Number of young per female Percentage females reproductively active Total reproduction days Growth (length and weight) | OECD declined to validate this assay | |
| OECD GD 201: New Guidance Document on Harpacticoid Copepod Development and Reproduction Test with Amphiascus | 5 |
Mortality Time to release of first and second clutch Number of necrotic/infertile eggs per clutch Morphological abnormalities Intersexuality Population growth rate Sex ratio | ||
| Daphnia Multigeneration Assay (when TG is available) | 5 |
Mortality Age at first reproduction Clutch size Intrinsic population growth rate Number of dead/aborted embryos/neonates Sex ratio | ||
| Insecta | OECD TG 218: Sediment–Water Chironomid Toxicity Test Using Spiked Sediment | 4 |
Mortality (including development and emergence) Time to emergence (males and females) Growth (weight) Sex ratio | Chironomus are protandrous so care must be taken when interpreting sex ratio changes |
| OECD TG 219: Sediment–Water Chironomid Toxicity Test Using Spiked Water | ||||
| OECD GD 239 Guidance Document on Honey Bee (Apis mellifera) Larval Toxicity Test, Repeated Exposure | 4 |
Larval mortalities from day 3 to day 8 Pupal mortalities from day 8 to day 15 Emergence rate on day 22 | ||
|
OECD TG 228: Determination of Developmental Toxicity to Dipteran Dung Flies (Scathophaga stercoraria L (Scathophagidae), Musca autumnalis De Geer (Muscidae)) | 4 |
Mortality Development Emergence (males and females) | ||
|
OECD TG 233: Sediment–Water Chironomid Life-Cycle Toxicity Test Using Spiked Water or Spiked Sediment | 5 |
Mortality Development Emergence Time to emergence (males and females) Growth (weight) Reproduction Fertility Sex ratio | ||
| Mollusca | OECD TG 242: Potamopyrgus antipodarum Reproduction Test | 4 |
Mortality Reproduction (No. of embryos) | |
| OECD TG 243: Lymnaea stagnalis Reproduction Test | 4 |
Mortality Reproduction (No. of egg clutches) | ||
| Nematoda | ISO 10872:2020: Determination of the toxic effect of sediment and soil samples on growth, fertility and reproduction of Caenorhabditis elegans (Nematoda) | 5 |
Survival Growth Reproduction |