P - Sea Alarm

Biomonitoring human populations after oil spills: the
Prestige experience
Blanca Laffon
Since 1960 → 410 tankers
377,765 tons of oil
PLACE
10th
Nº OF SPILLS
Gulf of Mexico
267
Northeast USA
140
Mediterranean Sea
127
Persian Gulf
108
North Sea
75
Japan
60
Baltic Sea
52
United Kingdom and English Channel
49
Singapur and Malasia
39
West coast of France and Northwest of Spain
33
Corea
32
Last 30 years:
- Polycommander (1970)
- Urquiola (1976)
- Andros Patria (1978)
- Aegean Sea (1992)
- Prestige (2002)
Oil Tanker
Year
Place
Spilled oil (Tm)
1
Atlantic Empress
1979
Tobago
280.000
2
ABT Summer
1991
700 miles from Angola
260.000
3
Castillo de Bellver
1983
Saldanha Bay, South Africa
257.000
4
Amoco Cádiz
1978
French coast of Bretagne
227.000
5
Haven
1991
Geneva, Italy
140.000
6
Odyseey
1988
700 miles from New Scotland
132.000
7
Torrey Canyon
1967
Scilly islands
119.000
8
Urquiola
1976
A Coruña, Spain
108.000
9
Hawaiian Patriot
1977
300 miles from Honolulu
99.000
10
Independenta
1979
Bosphorus, Turkey
93.000
11
Braer
1993
Shetland islands
85.000
12
Khark 5
1989
120 miles from Atlantic coast of Morocco
80.000
13
Jakob Maersk
1975
Oporto, Portugal
80.000
14
Aegean Sea
1992
A Coruña, Spain
72.000
15
Katina P
1992
Maputo, Mozambique
72.000
16
Nova
1985
20 miles from Iran
70.000
17
Prestige
2002
A Coruña, Spain
66.000
18
Wafra
1971
Cape Agulhas, South Africa
65.000
19
Assimi
1983
55 milles from Muscat, Oman
53.000
20
Metula
1974
strait of Magallanes, Chile
53.000
21
Exxon Valdez
1989
Prince William Sound, Alaska
37.000
Prestige accident: November 2002, 130 miles from Galicia
Content: 77,000 Tm
Spill: ≈44,000 Tm
42º 12.6’ N, 12º 3.8’ W
Potentially Protected Sea
Area
Next weeks:
22,000 Tm
Next months:
125 tons/day
900 Km
Nov 2002
Jul 2003
327,476
volunteers
Prestige oil
Heavy oil No. 2 (AFNOR Normative NF M 15-010 to 15-013)
Oil No. 6 (USEPA)
Mixture of compounds resulting from refining of crude oil,
very viscose and water insoluble
•VOC
Commission Directive 93/67/EEC
•PAH
Council Regulation 793/93
•Heavy metals
Commission Regulation 1488/94
Skin
- Exposure routes:
Respiratory tract
Digestive tract
- Acute toxicity:
Negative influence on psychology, immunology and
development of several organisms
Headaches, respiratory and eye irritations, sickness,
nauseas, etc.
Genotoxicity
- Chronic toxicity:
Chemical agent
Metastatic cancers
Anxiety, depressions, posttraumatic
stress, etc.
DNA
Partially transformed cells
Pre
tic
s
a
pl
o
e
n
es
g
a
st
VOC
Anthropogenic sources
High volatility in
Toxic, some have mutagenic and/or
carcinogenic properties (styrene, benzene,
etc.)
natural conditions
Human carcinogen (IARC)
CH3
Cytotoxic
CH3
CH3
CH3
CH3
Haematotoxic
Immunotoxic
CH 3
CH3
Benzene=59mg/kg
BTEX=345mg/kg
PAH
Use of fossil fuels
Genotoxic effects (mutagenic, carcinogenic and
teratogenic)
Benzo(a)anthracene
Benzo(a)pyrene
Benzo(b)fluoranthene
Dibenzo(ah)anthracene
Benzo(k)fluoranthene
Indene(1,2,3-cd)pyrene
7,8-diol-9,19-epoxide
Depends on metabolic
activation
Classified as possible or
probable human
carcinogens (IARC)
16 priority
contaminants (USEPA)
Heavy metals
Directly bind to DNA
Epigenetic alterations
DNA-DNA crosslinks
DNA-protein crosslinks
Oxidative DNA
damage
Cadmium
Aluminium
Group 1
(IARC)
Nickel
Lead
Group 2B
(IARC)
Interference
with DNA
repair systems
Objectives:
1. To evaluate the genotoxic effect induced by oil exposure during handling of oilcontaminated birds and cleaning beaches and rocks affected by Prestige spill.
2. To determine the influence of physiologic factors, consumption habits and use of
protective devices on the genotoxicity biomarkers evaluated.
OUR STUDY
¾ 1st part: individuals who performed autopsies and cleaning of
oil-contaminated birds
¾ 2nd part: individuals who participated in the
cleaning of beaches and rocks
34 exposed
71
individuals
35 controls
12 1-hour air samples in 2h intervals
Environmental VOC
Comet assay
Genotoxic effects
MN test
1st part: oil-contaminated birds
Exposure levels
VOC
Concentration (µg/m3 )
250
TVOC
BTEX
200
150
100
50
0
10:45
14:45
18:45
22:45
Time (h)
2:45
6:45
1st part: oil-contaminated birds
Biomarkers of effect
Comet assay
r=0.376, P<0.05
**
Exposed
60
Controls
50
43
44
45
TL (µM)
46
47
48
**
150-500h
>500h
**
TL (µm)
42
**
40
30
control
<150h
Time of exposure
**P≤0.01, significant difference with regard to controls and all other exposed groups
1st part: oil-contaminated birds
Biomarkers of effect
MN test
MN frequency (per 1000 cells)
30
Exposed
Controls
20
10
0
14
14.5
15
15.5
16
16.5
17
17.5
control
MN frequency
Chronic exposures
-Cytogenetic tests: cumulative damage
-Comet assay: recent exposures, damage easily repaired
<150h
150-500h
Time of exposure
>500h
1st part: oil-contaminated birds
Biomarkers of effect
Sex
Comet assay
Controls
Exposed
N
TL±SE
N
TL±SE
Male
18
43.42±0.10
18
47.86±0.17
Female
17
44.50±0.11
16
45.61±0.21**
**P≤0.01, significant difference with regard to males
MN test
Controls
Exposed
N
MN±SE
N
MN±SE
Male
18
14.22±0.73
18
16.61±0.71
Female
17
16.18±1.12
16
15.87±1.08
1st part: oil-contaminated birds
Biomarkers of effect
Age
r=0.464, P<0.01
Comet assay
Controls
N
TL±SE
Exposed
N
TL±SE
≤25 years
29 43.91±0.08
15
45.12±0.23
>25 years
6
19
48.14±0.15**
44.13±0.18
**P≤0.01, significant difference with regard to the minor-age group
MN test
Controls
Exposed
N
MN±SE
N
MN±SE
≤25 years
26
15.00±0.78
15
15.53±1.07
>25 years
9
16.00±1.39
19
16.84±0.73
1st part: oil-contaminated birds
Biomarkers of effect
Smoking
Comet assay
Controls
Exposed
N
TL±SE
N
TL±SE
Non-smokers
24
44.04±0.08
23
47.20±0.17
Smokers
11
43.75±0.14**
11
45.97±0.23**
**P≤0.01, significant difference with regard to non-smokers
MN test
Controls
Exposed
N
MN±SE
N
MN±SE
Non-smokers
24
15.00±0.82
23
16.43±0.80
Smokers
11
15.54±1.28
11
15.91±1.01
1st part: oil-contaminated birds
Biomarkers of effect
Protective devices
Comet assay
50
49
**P≤0.01, significant
difference with regard to
the use of mask
**
TL (µm)
48
47
46
45
44
43
Without Mask
With Mask
Without Mask
With Mask
MN test
MN frequency
18
17
16
15
OUR STUDY
¾ 2nd part: individuals who participated in the
cleaning of beaches and rocks
62 volunteers (V)
258
individuals
5 days – 4h/day
78 manual workers (MW)
4 months - 6.5h/day
60 workers using high pressure machines (HPW)
60 controls
Individual dosimeters
Environmental VOC
1-OHP and Σ-OHF
Urinary metabolites of PAH
Al, Cd, Ni, Pb and Zn
Blood heavy metals
Comet assay
Genotoxic effects
MN test
SCE
3 months –
6.5h/day
tra
MW
500
400
300
200
100
b
ch enz
e
lo
ro ne
et
he
n
to e
lu
e
nn
he e
pt
a
n- ne
et oct
a
hy
lb ne
en
z
m
+p ene
-x
yle
o- ne
xy
le
n
is
op
st e
yr
ro
e
py
lb ne
en
ze
ne
a
np
pr
o p i ne
ilb ne
1,
en
3,
ce
5n
t ri
b
m
-p o
et
in
hy
e
l b no
1,
en
2,
z
4n- ene
t ri
m
de
et
hy can
lb
e
en
ze
ne
600
tra
600
Concentration (µg/m 3)
te
t ra b
ch e n
lo ze
ro ne
et
he
to n e
n- lue
he ne
pt
a
n
et -o ne
hy ct
l b an
e
e
m nz
+p en
-x e
yl
o- ene
x
is
yl
op
e
ro st ne
p y yr
lb en
en e
ze
na
pr
-p ne
o
pi ine
1,
lb
3,
en ne
5c
tri
m b en
et -p o
hy in
1,
lb en
2,
e
o
41, trim n nze
2,
n
3- et -de e
tri hy ca
m lb
et en ne
hy z
lb en
en e
ze
ne
Concentration (µg/m 3)
VOC
te
b
ch enz
lo
e
ro n e
et
he
n
to e
lu
e
nn
he e
pt
a
n- n e
et oct
hy
a
lb ne
en
z
m
+p ene
-x
yl
e
o- ne
xy
le
is
ne
op
s
t
ro
yr
py
e
lb ne
en
ze
ne
a
npr -pi
o p ne
ilb ne
1,
en
3,
ce
5tri
n
m b -p o
et
hy inen
lb
1,
en o
2,
ze
41, trim n-d ne
2,
ec
3- eth
an
y
tri
m l be e
n
et
hy z e
n
lb
en e
ze
ne
te
Concentration (µg/m 3)
2nd part: beaches and rocks
Exposure levels
Profiles
V
500
400
300
200
100
0
600
0
HPW
500
400
300
200
100
0
2nd part: beaches and rocks
Exposure levels
VOC Differences between exposure groups
V
MW
HPW
3
Concentration (µg/m )
600
400
*
200
*
*
**
*
**
0
Total VOC
BTEX
Benzene
**P≤0.01, *P≤0.05, significant difference with regard to V group
2nd part: beaches and rocks
Exposure levels
PAH
1-OHP
N
Initial
Mean
Final
mean
Difference
between means
P-value
%change
V
45
0.0629
0.0873
0.0244
0.050*
38.79
MW
66
0.0941
0.1154
0.0213
0.205
22.63
HPW
24
0.1050
0.0771
-0.0279
0.227
-26.57
*P≤0.05, significant difference with regard to initial values
Σ-OHF
N
Initial
Mean
Final
mean
Difference
between means
P-value
%change
V
45
0.5069
0.7858
0.2789
0.008**
55.02
MW
66
0.9788
1.2847
0.2773
0.057
27.53
HPW
24
0.8325
0.7417
-0.0908
0.549
-10.91
**P≤0.01, significant difference with regard to initial values
2nd part: beaches and rocks
Exposure levels
1.2
Aluminium
**
Aluminium (µg/l)
20
Cadmium (µg/l)
Heavy metals
1
0.8
0.6
0.4
0.2
15
0
10
**
Controls
*
80
5
0
V
MW
HPW
Nickel
Lead (µg/l)
8
V
MW
HPW
Lead
**
60
Controls
40
20
**
6
0
Controls
V
MW
HPW
4
2
50
**
0
Controls
V
MW
HPW
**P≤0.01, *P≤0.05, significant difference with
regard to controls
Zinc
40
Zinc (µg/l)
Nickel (µg/l)
Cadmium
**
**
30
20
10
Controls
V
MW
HPW
2nd part: beaches and rocks
Biomarkers of effect
Comet assay
HPW
**
MW
**
V
**
Exposed
**
Controls
0
0.1
0.2
%TDNA
**P≤0.01, significant difference with regard to controls
0.3
0.4
2nd part: beaches and rocks
Biomarkers of effect
MN-BNMN
HPW
MW
**
V
Exposed
Controls
0
3
6
9
BNMN
**P≤0.01, significant difference with regard to controls
12
2nd part: beaches and rocks
Biomarkers of effect
Results similar in 1st and 2nd parts:
-significant ↑ comet assay
-no significant effect in MN frequencies
MN-CBPI
HPW
**
MW
**
V
Exposed
Controls
1.2
1.3
CBPI
**P≤0.01, significant difference with regard to controls
1.4
2nd part: beaches and rocks
Biomarkers of effect
SCE
HPW
**
MW
V
Exposed
Controls
3
3.2
3.4
3.6
3.8
SCE
**P≤0.01, significant difference with regard to controls
4
2nd part: beaches and rocks
Biomarkers of effect
SCE
4
3
2
1
PRI
HFC
HP
W
M
W
V
Ex
po
se
d
Co
nt
ro
ls
0
2nd part: beaches and rocks
Sex effect
Controls
Exposed
V
MW
HPW
Sex
N
%TDNA
N
BNMN
N
SCE
Male
23
0.080±0.003
25
6.76±0.95
16
3.74±0.07
Female
37
0.078±0.002
37
6.92±0.68
26
3.78±0.05
Male
88
0.166±0.003
79
6.46±0.48
35
3.51±0.04
Female
82
0.191±0.004**
78
6.96±0.50
33
3.85±0.04**
Male
19
0.364±0.011
21
3.14±0.39
10
3.25±0.08
Female
33
0.302±0.007**
37
4.84±0.43*
13
3.53±0.07**
Male
39
0.099±0.004
34
8.44±0.84
14
3.58±0.07
Female
20
0.110±0.004**
19
0.37±1.20
38
3.70±1.00
Male
30
0.127±0.003
24
6.54±0.64
11
3.62±0.07
Female
29
0.115±0.003**
22
7.59±0.86
12
4.28±0.08**
**P≤0.01, *P≤0.05, significant difference with regard to males.
2nd part: beaches and rocks
Age effect
Controls
Exposed
V
MW
HPW
Age
(years)
N
%TDNA
N
BNMN
N
SCE
17-21
20
0.085±0.003
20
7.85±1.23
16
3.82±0.07
22-34
35
0.078±0.002*
37
7.81±0.94
22
3.79±0.06
>34
3
0.048±0.004**
4
5.67±0.33
2
3.82±0.20
17-21
26
0.280±0.008
27
4.07±0.47
17
3.49±0.07
22-34
72
0.195±0.003**
66
6.44±0.56
23
3.63±0.05
>34
64
0.119±0.003**
57
9.79±0.73**
20
3.81±0.06**
17-21
19
0.336±0.010
21
3.62±0.45
13
3.46±0.07
22-34
29
0.308±0.008
32
4.94±0.52
9
3.33±0.08
>34
3
0.417±0.031*
3
5.33±2.40
1
3.52±0.18
17-21
3
0.113±0.010
3
6.33±2.85
5
2.82±0.14
22-34
21
0.101±0.004
18
8.17±1.15
2
3.74±0.12**
>34
33
0.099±0.003
31
11.45±1.16
9
3.58±0.08**
17-21
4
0.111±0.008
4
4.00±0.91
2
4.69±0.27
22-34
22
0.126±0.004
13
6.00±0.70
9
3.85±0.09
>34
28
0.115±0.003
25
9.12±0.94
9
4.07±0.08
**P≤0.01, *P≤0.05, significant difference with regard to the minor age group.
2nd part: beaches and rocks
Smoking effect
- Comet assay
Non-smokers
%TDNA
0.4
≤10cig/day
>10cig/day
**
0.3
**
0.2
**
0.1
**
0
Controls
Exposed
V
MW
**P≤0.01, significant difference with regard to non-smokers
HPW
2nd part: beaches and rocks
Smoking effect
- MN (BNMN)
Non-smokers
≤10cig/day
>10cig/day
10
BNMN
8
6
4
2
0
Controls
Exposed
V
MW
**P≤0.01, significant difference with regard to non-smokers
HPW
2nd part: beaches and rocks
Smoking effect
- SCE
Non-smokers
5
≤10cig/day
>10cig/day
**
SCE
4.5
**
** **
4
*
3.5
3
Controls
Exposed
V
MW
**P≤0.01, significant difference with regard to non-smokers
HPW
2nd part: beaches and rocks
Protective devices
- Clothes
Exposed
V
MW
HPW
Protective
clothes
N
%TDNA
N
BNMN
N
SCE
New
35
0.234±0.006
32
6.09±0.74
17
3.99±0.06
Reused
91
0.156±0.003
79
7.87±0.52
29
3.90±0.05**
None
42
0.166±0.004
44
5.12±0.52
14
3.58±0.07**
New
10
0.311±0.009
12
3.25±0.91
8
3.21±0.82
Reused
3
0.303±0.013
4
3.00±0.41
2
2.89±1.17
None
40
0.346±0.011*
41
4.63±0.41
14
3.58±0.07**
New
15
0.128±0.008
15
8.60±1.20
6
3.20±0.08
Reused
40
0.082±0.012
35
9.00±0.89
9
3.75±0.07**
None
2
0.101±0.002
2
15.00±1.00
0
New
10
0.113±0.006
5
5.40±0.50
3
3.77±0.14
Reused
48
0.123±0.003
40
7.38±0.41
18
4.05±0.06
None
0
0
0
**P≤0.01, *P≤0.05, significant difference with regard to the use of new clothes.
2nd part: beaches and rocks
Protective devices
- Mask
Exposed
V
MW
HPW
Protective
mask
N
%TDNA
N
BNMN
N
SCE
New
57
0.204±0.005
56
5.97±0.40
22
3.50±0.05
Reused
96
0.173±0.004
83
7.91±0.49
34
3.78±0.05**
None
15
0.150±0.004
15
5.83±0.78
8
3.60±0.09
New
41
0.349±0.010
44
4.48±0.36
15
3.30±0.06
Reused
5
0.286±0.009
5
3.80±1.23
3
3.36±0.18
None
7
0.340±0.015
8
3.13±0.55
5
3.84±0.13**
New
44
0.106±0.004
5
11.60±3.20
3
4.19±0.19
Reused
47
0.067±0.007
43
8.86±0.75
12
3.43±0.07
None
5
0.104±0.003
4
8.75±2.56
2
3.31±0.15
New
11
0.114±0.005
7
4.86±0.60
4
3.93±0.13
Reused
5
0.127±0.003
35
7.69±0.64
17
4.08±0.07
None
41
0.078±0.007
0
6.33±0.88
1
2.88±0.24
**P≤0.01, *P≤0.05, significant difference with regard to the use of new mask.
RESEARCH GROUP:
Josefina Méndez, PhD
Eduardo Pásaro, PhD
Blanca Laffon, PhD
Beatriz Pérez-Cadahía, PhD
Julia García-Lestón, PhD student
Vanessa Valdiglesias, PhD student
Alba Piñeiro, technician