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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