Test system

Exposure method (exposure method, concentration, vehicle, solution exchange time, and the others)

Organism characteristics (age, developmental stages)

PFAA type

Exposure time (sampling time, refresh time, generation)

Effects

Fish

Fathead minnow (Pimephales promelas)

Flow-through exposure; 45 ml/min solution supply; 0, 0.03, 0.1, 0.3, 1 mg/L PFOS

F0: Six-to-seven-month adult fish

F1: From embryo to fry 24 dpf

PFOS

14 d (at 1 mg/L), 21 d, and 24 d

Reproductive toxicity, endocrine disruption, PFOS accumulation

Juvenile atlantic salmon

(Salmo salar)

Dietary exposure; 0.2 mg/kg PFOA, PFOS; 0.01% methanol solvent

Juvenile fish (sexually immature), average mass of 70 ± 20 g

PFOA, PFOS

Sampling at 0, 2, 5, 8, and 14 d 0, 3, 6 d food spike exposure and 7 d recovery

Beta-oxidation, oxidative stress

Rainbow trout (Oncorhynchus mykiss)

Dietary exposure; PFC mixture exposure (in the ratio of 1:1:1:1, mix A composed of 5 mg/L individual PFC (PFOA, PFNA, PFDA, PFUnDA), mix B composed of 50 mg/L individual PFC, mix C composed of 250 mg/L individual PFC). For VTG, PFOA and PFDA exposure, 0.026, 0.128, 0.64, 3.2, 16, 80, 400, 2000 mg/L PFOA or PFDA; DMSO vehile (≤0.05 mg/L)

Subchronic PFC exposure: 11 month old juvenile trout; PFOA and PFDA exposure for VTG analysis: 5 month old juvenile trout

PFOA, PFNA, PFDA, PFUnDA, PFPA, PFHxA, PFHpA, PFDoDA, PFTrDA, PFTDA, PFBS, PFOS, PFDS, E2

14 d (feeding every 5 d per a week)

Endocrine disruption

Zebrafish (Danio rerio)

Static-exposure; 0.1% DMSO(v/v); 8, 16, 32 µM PFOS for sensitive period screening; 16 µM PFOS for histological examination, transmission electron microscopic examination, NimbleGen microarray

Embryo within 0.5 h of spawning

PFOS

0 to 48 hpf or 48 - 96 hpf for sensitive period screening; 48 - 96 hpf for histological examination and transmission electron microscopic examination; from 48 hpf during 48 h for NimbleGen microarray

Developmental toxicity, microassay gene profiles

Zebrafish (Danio rerio)

Semi-static exposure (50% renewals every 5 d, except for initial 5 d when embryo hatching occur); 0.01% DMSO; 0.5 µM PFOS (250 µg/L)

8 hpf

PFOS

From 8 hpf to 120 dpf

Thryoid toxicity

Marine medaka (Oryzias melastigma)

Semi-static exposure (25‰ saline water); 1, 3, 10 µg/L PFBS; daily exchange

Eggs

PFBS

From egg state to sexual maturity state (6 month old)

Ocular toxicity, neuronal toxicity, protein profilings

Zebrafish (Danio rerio)

Semi-static exposure (50% renewals every 5 d, except for initial 5 d when embryo hatching occur); 0.01% DMSO; 0.5 µM PFOS

Developmental stage from 8 h dpf to 150 dpf

PFOS

5 month exposure (from 8 h dpf to 150 dpf), followed by 5 d spawning time, hatching, and others

Lipid metabolism change, ATP content, gross index, histological changes, endocrine disruption

Pseudokirchneriella subcapitata, Daphnia magna, Oncorhynchus mykiss

Acute toxicity test: Static exposure; APFO (PFOA + ${\text{NH}}_{\text{4}}^{\text{+}}$ salt); to D. magna, 0, 100, 178, 316, 562, 1000 mg/L APFO; to O. mykiss, 0, 31.3, 62.5, 125, 250, 500, 1000 mg/L APFO; to P. subcapitata, 5.76, 11.37, 22, 70, 46.33, 95.87, 180, 67, 369.67 mg/L APFO);

Chronic toxicity test: To D. magna (Semi-static exposure: 0, 4.31, 9.16, 20, 44.2, 88.6 mg/L); to O. mykiss (Flow-through exposure (5.76 replacement per day, ELS test, 0, 2.18, 4.48, 10.7, 20.9, 40 mg/L)-OECD 201, 202, 203; EU commission directive 92/69 EEC, OECD 210, 211

Acute test: 6 h - 24 h neonate D. magna, 40 mm - 50 mm O. mykiss, 104 cells/ml inoculation P. subcapitata, Chronic test: 6 h - 24 h neonate D. magna, Fertilized egg to 85 d O. mykiss

APFO (PFOA + ${\text{NH}}_{\text{4}}^{\text{+}}$ salt)

Acute toxicity test: D. magna: 48 h, O. mykiss: 96 h, P. subcapitata: 96 h;

Chronic toxicity test: D. magna 21 d, O. mykiss 85 d

Acute toxicity, reproductive toxicity(ex. 95 d-O. mykiss ELS test)

Zebrafish (Danio rerio)

Semi-static exposure, replaced 50% at every 5 days; 8 hpf to 180 dpf exposure period; 0.01% DMSO; 0, 0.02, 0.1, 0.5 µM PFOS

8 hpf embryo

PFOS

8 hpf to 180 dpf

Lipid metabolism disturbance, oxidative stress

Thicklip grey mullets

(Chelon labrosus),

Static exposure; 2 mg/L PFOS dissolved in seawater

Juvenile fish

PFOS

2 and 16 d

Transport gene expression change

European bullhead (Cottus gobio)

Semi-static exposure; a half of water was replenished every 24 h; 0.1 and 1 mg/L PFOS

Adult fish, 10.4 ± 4.1 g; catching organism in Samson river and acclimated in tap water

PFOS

96 h

Enzyme activity, protein expression change (ex.: HSPA4, HSC70-2 etc.)

Zebrafish

(Danio rerio)

Semi-static exposure, replenished half every day, 0.002% DMSO; 0, 10, 50, 250 µg/L PFOS

F0 fish: 14 dpf zebrafish fry

PFOS

F0 fish: 70 d PFOS exposure and 30 d only water exposure, (100 d)

F1 fish: Offspring of PFOS treated female and nonchemical treated male fish

F0: Gross index, thyroid toxicity, endocrine disruption F1: Reproductive toxicity endocrine disruption