acute toxicity of individual polycyclic aromatic hydrocarbons (pah) to early life stages of...
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Acute toxicity of individual polycyclic aromatic hydrocarbons (PAH) to early life stages of commercial marine organisms, zooplankton and phytoplankton
R. Beiras, J. Bellas, L. Saco-Álvarez, J.C. Mariño-Balsa, P. Pérez, N. Fernández
Universidade de Vigo, Laboratorio de Ecoloxía Mariña (LEM), Facultade de Ciencias do Mar , E-36310, Galicia.
(VEM2003-20068-C05-02)
Ecotoxicological evaluation
1. Reference toxicants: medium MW PAH
Phenathrene Fluoranthene Pyrene14 C 16 C 16 C178 MW 202 202435 µg/L solub. 260 133
-. Degradation products
1-OH Pyr
2. Water-accommodated fraction of the fuel (WAF)
3. Seawater from affected coastline
OH
R. Beiras et al. VERTIMAR Vigo
simplecontrolledartificial
unreproducibleenvironmentally relevant
still on progress
1. Ecotoxicological evaluation of medium MW (1) PAHs
(1) Low mollecular weight aromatics are too volatile and very high MW aromatics too insoluble (Neff & Stubblefield, 1995)
[fluoranthene] µg/l
control control acet 1 4 16 64 256
% b
iolo
gica
l re
spo
nse
0
20
40
60
80
100normal hatched larvaenon developed embryosabnormal larvae (alive)abnormal larvae (dead)
One week old Gadus morhua embryos exposed to fluoranthene for 7 days
[fluoranthene] µg/l
control control acet 1 4 16 64 256
% s
urv
iva
l
0
20
40
60
80
100
48 h 72 h
One day old Gadus morhua larvae exposed to fluoranthene for 72 h.
Fish embryos and larvae (cod)Dr. J. Bellas, Kristineberg Marine Station, Suecia
LOEC: 1 µg/L
LOEC: 64 µg/L
R. Beiras et al. VERTIMAR Vigo
? 64 µg/L
0
20
40
60
80
100
0 50 100 150 200 250 300
Fenantreno (µg l-1)
% S
uper
vive
ncia
48 h-OSC
48 h-LUZ
0
20
40
60
80
100
0 50 100 150 200 250 300
Fluoranteno [µg l-1]
% S
up
ervi
ven
cia
24 h- LUZ
24 h-OSC
Graf. 1 Graf. 2
Crustacean larvae (Palaemon serratus)Dr. J. C. Mariño-Balsa. Centro de Repoboación de Especies Mariñas, Muxía
R. Beiras et al. VERTIMAR Vigo
LIG
HT
DARK64 µg/L 32 µg/L
250 µg/L
Larvas de erizo - PAHsL. Saco-Álvarez, Universidade de Vigo.
-8.0 -7.5 -7.0 -6.5 -6.0 -5.5 -5.0 -4.50.00.10.20.30.40.50.60.70.80.91.01.11.2
LIGHT
DARK
EC50LIGHT 2.393e-007 EC50
DARK 1.445e-006
LOG [fluoranthene] (M)
larv
al g
row
th(%
con
tro
l) ***
R. Beiras et al. VERTIMAR Vigo
Flu
-7.0 -6.5 -6.0 -5.5 -5.0 -4.5 -4.00.00
0.25
0.50
0.75
1.00
1.25
LIGHT
DARK
EC50LIGHT 6.619e-006 EC50
DARK 8.515e-006
LOG [phe] (M)
larv
al g
row
th(%
con
tro
l)Larvas de erizo - PAHsL. Saco-Álvarez, Universidade de Vigo.
n.s.
R. Beiras et al. VERTIMAR Vigo
Phe
Larvas de erizo - PAHsL. Saco-Álvarez, Universidade de Vigo.
-8.0 -7.5 -7.0 -6.5 -6.0 -5.5 -5.0 -4.50.0
0.2
0.4
0.6
0.8
1.0
1.2
LIGHT
DARK
EC50LIGHT 4.275e-007 EC50
DARK 1.285e-006
LOG [pyr] (M)
larv
al g
row
th(%
con
tro
l) ***
R. Beiras et al. VERTIMAR Vigo
Pyr
Phenanthrene
[Phe] nM
0 395 791 1582 3164
Gro
wth
rat
e (d
ay-1
)
0.04
0.06
0.08
0.10
0.12
0.14
ANOVA: p<0.05
Dunnett test *
EC10=1169nM, 208 µg/L
EC50=4320nM, 768 µg/L
**
R. Beiras et al. VERTIMAR Vigo
Phytoplankton (Isochrysis galbana)P. Pérez, Univ. de Vigo
70 140 280 560 µg/L
Fluoranthene
[Phe] nM
0 173 346 692 1730
Gro
wth
rat
e (d
ay-1
)
0.04
0.06
0.08
0.10
0.12
0.14
*
ANOVA: p<0.05
Dunnett test *
R. Beiras et al. VERTIMAR Vigo
Phytoplankton (Isochrysis galbana)P. Pérez, Univ. de Vigo
35 70 140 280 µg/L
[Flu]
Pyrene
[Pyr] nM
0 74 148 297 593
Gro
wth
rat
e (d
ay-1
)
0.04
0.06
0.08
0.10
0.12
0.14
0.16 ANOVA: p>0.05
R. Beiras et al. VERTIMAR Vigo Phytoplankton (Isochrysis galbana)
P. Pérez, Univ. de Vigo
15 30 60 120 µg/L
Phenanthrene Fluoranthene Pyrene sp endpoint incubation parameter µg/L µM % sat µg/L µM %sat µg/L µM %sat
cod embryogenesis 7d, dark LOEC n.t. 64 0.32 24.6% n.t.urchin larval length 2d, dark EC10 90 0.51 20.7% 40 0.2 15.4% n.t.
larval growth 2d, dark EC10 n.c. 27 0.13 10.4% 70 0.34 52.6%2d, light EC10 113 0.63 26.0% 21 0.1 8.1% 23 0.12 17.3%
copep. egg production 2d, dark EC50 231 1.3 53.1% 87 0.43 33.5% 51 0.25 38.3%shrimp larval survival 3d, dark LC50 132 0.74 30.3% 94 0.47 36.2% >128
3d, light LC50 91 0.51 20.9% <32 <0.16 <12% >128phytop. growth rate 3d, light EC10 208 1.17 23.0% 99 0.49 38.1% >120
Summary
Conclusions on individual PAHs
-Medium MW PAHs cause toxicity on early life stages of marine organisms at concentrations in average 0,8 µM for Phe, 0,3 µM for Flu and 0,2 µM for Pyr, which corresponds to approx. 30% saturation for the three PAHs
-Light enhances the toxicity of Flu and Pyr but not Phe
2. Ecotoxicological evaluation of WAF
2. magnetic stirring (24 h) 3. GF filtration
1. mixing Prestige fuel oil with filtered seawater (40 g/L)
Preparation of water-accommodated fraction (WAF)
4. dilutions with FSW
Fish larvae (Cyprinodon variegatus)Dr. J. Bellas, I. Fernández-Méijome, Univ. de Vigo
WAF
control 12.5 25 50 100
% s
urvi
val
0
20
40
60
80
100
24 h 48 h 72 h 96 h
WAF
control 12.5 25 50 100
% s
urvi
val
0
20
40
60
80
100
24 h48 h72 h96 h
Luz Osc
WAF
control 25 37.5 50 75
% s
urvi
val
0
20
40
60
80
100
24 h48 h72 h96 h
WAF
control 25 37.5 50 75
% s
urvi
val
0
20
40
60
80
100
24 h48 h72 h96 h
Luz Osc
R. Beiras et al. VERTIMAR Vigo
Sea-urchin larvae -WAFI. Fernández Méijome, A. Méndez, Universidade de Vigo.
R. Beiras et al. VERTIMAR Vigo
20
40
60
80
100
-3,5 -3 -2,5 -2 -1,5 -1
Log dilution
Le
ng
th (
% C
on
tro
l)
Light Dark
WAF
Dilution
0 1/32 1/16 1/8
Gro
wth
rat
e (d
ay-1
)
0.040
0.045
0.050
0.055
0.060
0.065
**
***
***
ANOVA: p=0.000
Dunnett test *
Thalassiosira rotula
Phytoplankton (Thalassiosira rotula)P. Pérez, Univ. de Vigo
R. Beiras et al. VERTIMAR Vigo
dilution WAF
0.0 0.1 0.2 0.3 0.4 0.5 0.6
grow
th r
ate
%
0
20
40
60
80
100
120
p<0.001
R2=0.69
Thalassiosira rotula
R. Beiras et al. VERTIMAR Vigo
Findings on WAF
-Whilst the sensitivity of marine organisms to PAHs was very similar among phyla, large differences in sensitivity to WAF are appreciated, with urchins the most and fish the less sensitive
-Light moderately enhances the toxicity of WAF
-If we multiply the WAF dilutions shown above by the concentrations of medium MW PAHs measured in WAF from Prestige fuel oil (J.J. González et al.) the PAH concentrations obtained are orders of magnitude below the toxicity thresholds. Thus medium MW PAHs alone are not responsible for the toxicity of the WAF on marine organisms.
Summary
sp end point incubation parameter WAF dilution TU C. variegatus (fish) larval survival 4d, dark LD10 0.35 2.9
4d, light LD10 0.34 2.9P. lividus (urchin) larval growth 2d, dark ED10 0.0076 249
2d, light ED10 0.0042 131T. rotula (phytop.) growth rate 3d, light ED10 0.04 25
3. Ecotoxicological evaluation of environmental samples
Nov Dec Jan Feb Mar Apr May
larv
al le
ngth
(%
cont
rol)
0.2
0.4
0.6
0.8
1.0
1.2
M1 M2M3 ***
Nov Dec Jan Feb Mar Apr May
La
rval
leng
th (
%co
ntro
l)
0.0
0.2
0.4
0.6
0.8
1.0
1.2
M1 M2 M3
******
* **
**
Natural seawater from coastline affected by the oil-spillL. Saco Álvarez, Univ. de Vigo
Tox. in Nov.
Tox. in Nov. & Dec.
Dark Light
R. Beiras et al. VERTIMAR Vigo
M1: Furnas42º 38’ 38” N, 9º 02’ 15” W
Light-enhanced toxicity of M1 seawater sampled in Nov and Dec 2002 on Paracentrotus lividus sea-urchin embryos.
0%
20%
40%
60%
80%
100%
Site 21 Site 17 Site 13 Site C Site 5 Control
D-l
arva
e
R. Beiras et al. VERTIMAR Vigo
Sediment elutriates from coastline affected by the oil-spillDr. R. Beiras, Univ. de Vigo
0%
20%
40%
60%
80%
100%
0.00 0.50 1.00
Dilution
no
rma
l D-l
arv
ae
.
M1,1
M1,2
M1,3
Control
0%
20%
40%
60%
80%
100%
0,00 0,50 1,00
Dilution
no
rmal
D-l
arva
e.M2,1
M2,2
M2,3
Control
No toxicity of sediments from the Galician shelf (9 to 13th Feb 2003, IEO cruise) on V. pullastra clam embryos
No toxicity of M1 and M2 sand elutriates sampled Dec 17th on Crassostrea gigas oyster embryos.
Findings on natural samples
-We present direct evidence of toxicity of seawater taken the first few weeks from impacted sites on water-column organisms.
-In contrast the fraction of weathered fuel accumulated in the sand as solid particles and tar balls, visually much more conspicuous, lacked toxicity to water column organisms
Final Conclusions
-Except for phytoplankton (less sensitive), toxicity of Phe, Flu and Pyr is very similar for very different marine organisms, and it appears at concentrations around 30% saturation in seawater.
-The toxicity of the Prestige fuel oil WAF is much more selective (organism-specific) and it cannot be explained on the basis of its content in medium MW PAHs alone.
-Our findings stress the impact to water column organisms of the less conspicuous and frequently overlooked water-accommodated fraction, rather than the more visible oil slick