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Table 1 Internationally accepted invertebrate toxicity test guidelines. OECD CF level refers to the OECD Conceptual Framework [58]

From: Commentary: Assessing the endocrine disrupting effects of chemicals on invertebrates in the European Union

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
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
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
Emergence (males and females)
OECD TG 233: Sediment–Water Chironomid Life-Cycle Toxicity Test
Using Spiked Water or Spiked Sediment
5 Mortality
Time to emergence (males and females)
Growth (weight)
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
  1. Endpoints in bold are more indicative of a potential ED mode of action