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Table 2 Summary of selected standard tests where highlight is given to potential adaptations FOR THE TESTING OF MANUFACTURED NANOMATERIALS (including nanobiomaterials) of existing OECD/ISO guidelines FOR THE TESTING OF CHEMICALS regarding test duration and endpoints

From: Environmental hazard testing of nanobiomaterials

Test guideline Test endpoints Test duration (days)
Enchytraeus crypticus
Standard (OECD/ISO):
OECD 220; ISO 16386;
Survival and Reproduction 21–28
OECD 317 Kineticsa: uptake, elimination, BAFs 14 + 14
Alternative 1: A1
Full Life Cycle test (adapted from standard [55])
Hatching
Growth
Maturation
Survival
Reproduction
11
11–46
25
46
46
Alternative 2: A2
Extended test (adapted from standard [56])
Survival, Reproduction, Population growth 7, 14, 21, 28
60, 72, 84
Alternative 3: A3
Multigenerational test (adapted from standard [37, 57])
F1–F4 spiked soil + F5–F7 clean soil
Survival and reproduction; epigenetics 32, 64, 96, 128, 160, 192, 224
Recommendation:
Use A1, A2, A3
All 46–84
Implementation: Add A1, A2 and A3 in OECD/ISO as annexes.
Folsomia candida
Standard (OECD/ISO):
OECD 232;
ISO 11267;
Survival and Reproduction; 28
Alternative 1: B1
Multigenerational test (adapted from standard [58, 59])
F0, F1, F2: Survival, Reproduction, Size (adults, juveniles) 28, 56, 84
Recommendation: Use B1 All 28–84
Implementation: Add B1 in OECD/ISO as annex
Eisenia fetida
Standard (OECD/ISO):
OECD 222
Survival
Reproduction
28
56
OECD 317 Kineticsa: uptake, elimination, BAFs 21 + 21
Alternative 1: C1
Full Life Cycle (adapted from standard)
Hatching
Growth
Maturation
Survival
Reproduction
21–28
14, 28, 42, 56
60
14, 28, 42, 56
56
Alternative 2: C2
Tissue distribution and biochemical analysis
Tissue distributiona biomarkers, omics 3–7–14 (flexible)
Recommendation:
Use C1 and C2
All 28–84
Implementation: Add C1 in OECD/ISO as annex
Make use of C2 and proceed towards standardization
Use A1–A3 as surrogate (oligochaete with shorter life cycle)
Alternative 3: C3
Earthworm in vitro test [60]
Cell viability—biomarkers, omics, corona formation 1
Recommendation:
Use C3 for rapid mode-of-action understanding
All 1
Implementation: Further develop this system to a higher TRL level
Microorganisms
Standard (OECD/ISO):
OECD 216/ISO 14238
N-Transformation 28–100
Standard (OECD/ISO):
OECD 217/ISO 14239
C-Transformation 28–100
Alternative 1: D1
Fixed prolonged test duration
All microbial activities 84–100
Alternative 2: D2
ISO 15685
Potential ammonium oxidation 84–100
Recommendation: Use D1 and D2 All 84–100
Implementation: Add D1 in OECD/ISO as annex for particular materials
Add D2 in guidance for the testing of toxic ion releasing particular materials
Standard (OECD/ISO):
OECD 303A
C-Transformation, N-Transformation No fixed duration (plateau phase of min. 21 days)
Alternative 1: E1
Addition of denitrification chamber
C-Transformation, N-Transformation; sewage sludge for soil spiking Transformation: no fixed duration
Spiking: 10 days (mean sludge age in a wastewater treatment plant)
Recommendation: Use E1 C-Transformation, N-Transformation; sewage sludge for soil spiking Transformation: no fixed duration
Spiking: 10 days (mean sludge age in a wastewater treatment plant)
Implementation: Add E1 in OECD as annex for particular material
Daphnia magna
Standard (OECD/ISO):
OECD TG211
ISO 10706
Survival
Reproduction
Growth (length)
21 (daily monitoring)
Alternative 1: F1
Multigenerational (F0, F1, F2)
Survival
Reproduction
Growth (length)
21, 42, 70
Recommendation: Use F1 All 70
Implementation: Add F1 as annex to OECD TG211
Fish
Standard (OECD/ISO):
OECD TG305
Bioaccumulation Dietary: (10–14) + 28 or 42
Alternative 1: G1 Bioaccumulation Dietary: (10–14) + 52 or more
Recommendation: Use G1 All depuration period of 52 or more days
Implementation: Add G1 as annex to OECD TG305
  1. aInstrument and method that allows quantification of the specific material are required