1. Study on the behavior of the heavy metals
Cu, Cr, Ni, Zn, Fe, Mn and 137Cs
in an estuarine ecosystem using
Mytilus galloprovincialis as
a bioindicator species: the case of
Thermaikos gulf, Greece
Presented by :
Fiddy Semba Prasetiya
ECOMAMA, Vrije University of Brussels

2. Thermaikos GulfThermaikos Gulf

3. General IntroductionGeneral Introduction
 Anthropogenic activities :Anthropogenic activities :
- Sewage and Industrial discharges- Sewage and Industrial discharges
- Harbour activities- Harbour activities
- Agricultural activities- Agricultural activities
Pollution on marine ecosystemPollution on marine ecosystem

4. Marine pollutionMarine pollution
Strategy and routine monitoring of contaminantStrategy and routine monitoring of contaminant
in marine environmentin marine environment
Mussels as pollution bio-indicatorMussels as pollution bio-indicator

5. Benthos as pollution bio-indicatorBenthos as pollution bio-indicator
 Benthos as pollutionBenthos as pollution
bio-indicatorbio-indicator (Regoli, 1998)(Regoli, 1998)
 Mytilus galloprovincialisMytilus galloprovincialis
as a local bio-indicatoras a local bio-indicator
(Farrington(Farrington et al.et al., 1983), 1983)

6. Objective of the studyObjective of the study
 To elucidate the seasonal behavior of metalsTo elucidate the seasonal behavior of metals
(Cu, Cr, Ni, Zn, Fe, Mn and(Cu, Cr, Ni, Zn, Fe, Mn and 137137
Cs) in musselsCs) in mussels
from an estuarine ecosystem,from an estuarine ecosystem,
 To provide information on the interrelation ofTo provide information on the interrelation of
metals, as well as to confirm the use ofmetals, as well as to confirm the use of
cultivatedcultivated MytilusMytilus galloprovincialisgalloprovincialis as bio-as bio-
indicator for heavy metals andindicator for heavy metals and 137137
Cs pollutionCs pollution

7. MethodologyMethodology
 Sampling:Sampling:
-- 94 composite samples of94 composite samples of M. galloprovincialisM. galloprovincialis (82(82
for metals and 12 forfor metals and 12 for 123123
Cs) collected from stat.LOUCs) collected from stat.LOU
and stat.XAand stat.XA
-- Sampling period: April to October 2000 (monthly)Sampling period: April to October 2000 (monthly)

8. Location of sampling stationsLocation of sampling stations

9. MethodologyMethodology
 Heavy metal analyses:Heavy metal analyses:
- A total of 1640 mussel specimens were examinedA total of 1640 mussel specimens were examined
- Five to six replicates of composite samples consisting of theFive to six replicates of composite samples consisting of the
soft parts 20 specimens of similar size were preparedsoft parts 20 specimens of similar size were prepared
- Each month the size of the specimens was similar, butEach month the size of the specimens was similar, but
differed inter-monthly (with length varying from 3.5 – 7.0 cm)differed inter-monthly (with length varying from 3.5 – 7.0 cm)
- After freeze-drying with a CHRIST GAMMA 1-20After freeze-drying with a CHRIST GAMMA 1-20
lyophilisator, 1 g of dried tissue was digested with 10 mllyophilisator, 1 g of dried tissue was digested with 10 ml
HNOHNO33
- Dry weight was 22-25% of the fresh weight. After digestion,Dry weight was 22-25% of the fresh weight. After digestion,
sample was diluted with distilled water to 20 mlsample was diluted with distilled water to 20 ml
- A VS AA 20 plus flame AAS was used to determine Cu, Cr,A VS AA 20 plus flame AAS was used to determine Cu, Cr,
Ni, Zn, Fe and Mn concentrationsNi, Zn, Fe and Mn concentrations

10. MethodologyMethodology
 123123
Cs analyses:Cs analyses:
- Composite samples of at least 80-200 specimens (totalComposite samples of at least 80-200 specimens (total
weight about 8-10 kg each sample) were prepared byweight about 8-10 kg each sample) were prepared by
adding soft partsadding soft parts
- Each month the size of the specimens was similar, butEach month the size of the specimens was similar, but
differed inter-monthly (with length varying from 2.3 – 8.5 cm)differed inter-monthly (with length varying from 2.3 – 8.5 cm)
- Samples were ashed using the “dry ashing procedure” forSamples were ashed using the “dry ashing procedure” for
caesium and natural radionuclide measurementcaesium and natural radionuclide measurement
- The “ashing factor” factor was 7.1-14.6% of the fresh weightThe “ashing factor” factor was 7.1-14.6% of the fresh weight
- A quantity of 14-45 g of the ash sample was measured byA quantity of 14-45 g of the ash sample was measured by
gamma-spectroscopy measurementgamma-spectroscopy measurement

11. Results and discussionResults and discussion
 The levels of the measured metals, both radioactive and
stable, are characterized in general as low and
comparable to those from non-polluted Mediterranean
areas
 The analyzed samples showed lower concentrations of
Cu and Cr and similar or lower concentrations of Zn, Fe
and Mn when compared to the respective ones from Italy
 Mussels from Turkey had similar levels of Cu, Cr, Fe and
Mn and higher levels of Zn and Ni (Koklu et al., 2000; Topcuoglu
et al., 2002)

12. Results and discussionResults and discussion
Table 2. Summary statistic of contaminants in mussels fromTable 2. Summary statistic of contaminants in mussels from
Thermaikos gulf (in mg/kg dry weight for metals and Bq/kg wetThermaikos gulf (in mg/kg dry weight for metals and Bq/kg wet
weight forweight for 123123
CsCs
NN
AverageAverage
STDSTD
MedianMedian
MinMin
MaxMax
CuCu CrCr NiNi ZnZn FeFe MnMn 123123
CsCs
8282
3.913.91
1.291.29
3.553.55
1.671.67
6.356.35
8282
1.851.85
0.770.77
1.731.73
0.200.20
3.463.46
8282
4.344.34
1.391.39
4.584.58
1.161.16
7.517.51
8282
56.356.3
27.427.4
59.659.6
20.820.8
115.0115.0
8282
395395
159159
403403
8181
715715
8282
14.114.1
4.74.7
12.712.7
7.27.2
25.325.3
8282
1.231.23
0.480.48
1.201.20
0.30.3
1.901.90

13. Results and discussionResults and discussion
Table 3. Concentration of metals in mussels from Mediterranean and Black SeaTable 3. Concentration of metals in mussels from Mediterranean and Black Sea
AreaArea CuCu CrCr NiNi ZnZn FeFe MnMn
BulgariaBulgaria
PortugalPortugal
N.ItalyN.Italy
ItalyItaly
SpainSpain
SpainSpain
BosporusBosporus
Black SeaBlack Sea
UKUK
0.5-10.60.5-10.6
6.2-13.46.2-13.4
6.9-33.76.9-33.7
0.53-5.960.53-5.96
1.1-31.1-3
2.97-5.652.97-5.65
7.21-9.17.21-9.1
15.1-40.915.1-40.9
5.2-31.65.2-31.6
0.18-6.520.18-6.52
0.370.37
1.80-2.011.80-2.01
<0.06-7.58<0.06-7.58
0.6-5.70.6-5.7
1.3-111.3-11
4.02-24.074.02-24.07
5.0-42.95.0-42.9
12.8-42.012.8-42.0
140-542140-542
139-381139-381
10.8-5910.8-59
17.3-75.817.3-75.8
31.9-45.431.9-45.4
78-51278-512
110-368110-368
305-1175305-1175
11.7-18011.7-180
151-598151-598
215-1072215-1072
9.8-37.89.8-37.8
1.1-14.21.1-14.2
5.7-22.85.7-22.8
8.6-27.08.6-27.0
dwdw
dwdw
dwdw
wetwet
wwww
wetwet
dwdw
dwdw
dwdw

14. Results and discussionResults and discussion
 Levels of Cu, Cr, Ni and Zn were generally statisticallyLevels of Cu, Cr, Ni and Zn were generally statistically
lower (compared to Saronikos and Amvrakikos Gulf andlower (compared to Saronikos and Amvrakikos Gulf and
Larymna Bay)Larymna Bay)
 Saronikos gulfSaronikos gulf high levels of Cu and Znhigh levels of Cu and Zn
 Larymna bayLarymna bay high levels of Cr, Ni and Fehigh levels of Cr, Ni and Fe
 Thermaikos gulfThermaikos gulf high levels of Fe and Mnhigh levels of Fe and Mn

15. Fig.2 Mean values and LSD confidence intervals of metals in
mussels from diverse Greek areas
AMVR = Amvrakikos gulf,
LAR = Larymna bay,
SAR = Saronikos gulf,
TH = Thermaikos gulf-current data from both sampling stations (Catsiki, 2005)

16. Results and discussionResults and discussion
Time period (y)Time period (y) 123123
Cs (Cs (Bq/kg ww)Bq/kg ww) RemarksRemarks
1984 – 19851984 – 1985
1986 – 19871986 – 1987
1988 – 19951988 – 1995
1999 – 20011999 – 2001bb
LLDLLD
0.680.68 ± 0.10± 0.10
7.10 ± 1.807.10 ± 1.80
0.46 ± 0.280.46 ± 0.28
0.92 ± 0.680.92 ± 0.68
0.10.1
Excluded max.observed 3 samplesExcluded max.observed 3 samples
from one stationfrom one station
Mean valueMean value ± Standard deviation of± Standard deviation of
(present study)(present study)
Measurement in dried soft tissueMeasurement in dried soft tissue
transformed into wet weighttransformed into wet weight
Table 4.Table 4. 123123
Cs activity concentrations (Bq/kg wet weight) inCs activity concentrations (Bq/kg wet weight) in Mytillus galloprovincialisMytillus galloprovincialis in the Eastern Mediteranean –in the Eastern Mediteranean –
Aegean and Ionian Sea, (FlorouAegean and Ionian Sea, (Florou et al.,et al., 2002)2002)
 Levels ofLevels of 123123
Cs inCs in M. galloprovincialisM. galloprovincialis from Amvakikos gulf : 1.40from Amvakikos gulf : 1.40 ± 0.29 Bq/kg ww (slightly± 0.29 Bq/kg ww (slightly higher if compared to Italy andif compared to Italy and
Adriatic coasts)Adriatic coasts)

17. Fig. 3. Temporal evolution
of metals (average of five
to six replicates per
sampling) and 137Cs
activity concentrations of
composite samples) in
mussels
 2 groups of temporal
pattern in mussels:
- Cs-Ni-Fe with min.
during spring (April)
and autumn (October)
- Cu-Zn & Mn-Cr with
max. during spring
(April) and autumn
(October)

18. Temporal patterns of heavy metals
 2 groups of
temporal pattern in
mussels:
- Cs-Ni-Fe with min.
during spring
(April) and autumn
(October)
- Cu-Zn & Mn-Cr
with max. during
spring (April) and
autumn (October)