Evaluation of heavy metal and persistent organic pollutants input … · 2011. 9. 12. · Crisen benzo(k)fluoranthene Benzo[a]pyrene Benzo(ghi)perilen Dibenzo[a,h]anthracene Indeno(1,2,3-c,d)piren.

EVALUATION OF HEAVY METAL AND PERSISTENT

ORGANIC POLLUTANTS INPUT FROM DANUBE

WATERS IN THE BLACK SEA AREA INFLUENCED BY

THE DANUBE, IN THE CONTEXT OF RECENT

CLIMATIC CHANGES AND BIOACCUMULATION

OF THESE COMPOUNDS IN BIOTA

Valentina COATU, Victoria PIESCU, Andra OROS, R. MIHNEA

National Institute for Marine Research and Development

„Grigore Antipa” Constantza

ABSTRACT

The paper presents data regarding the level of heavy metals,

organochlorine pesticides and polycyclic hydrocarbons pollution

and their bioaccumulation in organisms from Danube mouth and

Black Sea area influenced by Danube.

The distribution of heavy metal, organochlorine pesticides and

polycyclic hydrocarbons concentrations underlined the phenomena

of pollutants concentration both in water and sediments, especially

in the eastern and southern part of the Musura Bay and in Sulina

and Sfântul Gheorghe arms outfalls.

The bioaccumulation of heavy metals and organochlorine

pesticides is more obviously in mollusks when compare with fish

species, but bioaccumulation of polycyclic hydrocarbons is similar

in both categories of organisms.

KEY WORDS: organochlorine pesticides, polycyclic

hydrocarbons, heavy metals, Danube mouth, Black Sea

INTRODUCTION

Despite the efforts and the results obtained by International

Commission for the Protection of the Danube River there are still rezones of

concern regarding Danube’s ecosystem state; one of them refers to dangerous

substances which are present in concentrations that represent a risk for aquatic

environment and human health.

Recent climatic changes had as result some disasters like huge

inundations. As a consequence, there were large quantities of waters

discharged in Danube river, waters that washed out large areas with various

uses and carried along considerable quantities of pollutants. Finally, this

pollutants flows into the Black Sea.

This paper presents data on heavy metals, organochlorine pesticides

and polycyclic hydrocarbons pollution and their bioaccumulation in organisms

from Danube mouth and Black Sea area influenced by Danube in 2006 – 2007.

MATERIAL AND METHODS

Water and sediment samples were collected from Danube arms: Chilia,

Sulina and Sfântul Gheorghe, Musura Bay and Black Sea area in front of

Sulina and Sfântul Gheorghe arms (Fig. 1).

Biota was represented by mollusks and fish species collected as a result

of research expeditions from marine area influenced by Danube between Sulina

and Portita and from fishermen in Sulina and Sfântul Gheorghe.

Samples were analyzed for heavy metals, organochlorine pesticides

and polycyclic aromatic hydrocarbons.

Organic pollutants were extracted from water samples in a separation

funnel using a mixture of 7/3 hexane/dichloromethane. Sediment and biota

samples were extracted 8 hours with hexane in Soxhlet apparatus.

The extracts were concentrated to about 20 ml in a rotary evaporator;

the sulphur compounds were removed with copper powder in an ultrasonic

bath (sediment samples) and lipids were removed with sulphuric acid (biota

samples); the samples were fractionated on alumina-silica gel columns for

polycyclic aromatic hydrocarbons and on florisil columns for organochlorine

pesticides; the final concentration was done in Kuderna-Denish concentrators

and in nitrogen stream up to 1 ml.

The detection of polycyclic aromatic hydrocarbons concentration in

extracts obtained as described above was done with a Perkin-Elmer gas

chromatograph with mass selective detector and for organochlorine pesticides

with Perkin-Elmer gas chromatograph with electron capture detector.

Fig. 1 - Sampling sites for water and sediments on Chilia, Sulina and Sfântul Gheorghe arms,

Musura Bay and marine area in front of Sulina and Sfântul Gheorghe arms:

1- Chilia arm; 2 – Tulcea - Sulina arm; 3 – Crişan - Sulina arm; 4 – Sulina – Sulina arm;

5 – Murighiol - Sfântul Gheorghe arm; 6 – Musura Bay N; 7 – Musura Bay V; 8 – Musura

Bay S; 9 – Musura Bay E; 10 - Sulina discharging area S; 11- Sfântul Gheorghe dischargind

area N; 12- Sfântul Gheorghe discharging area S.

For heavy metals analyse unfiltered water samples, preserved in plastic

bottles in order to avoid contamination, were acidified to pH 1.5 – 2.0 with

HNO3 and stored at about 40C. Total metal concentrations were analysed

directly, by atomic absorption spectrometry method, using an ATI-UNICAM

939Z spectrometer, with graphite furnace and Zeeman background correction

device.

Sediments samples were oven-dried at 1050C and well homogenized (>

2 mm fraction removed). Dried sediment and biota (fresh) samples were

weighted and treated with 5 ml concentrated nitric acid 65%, HNO3 Suprapur

(Merck).

Digestion procedure was carried out in a Microwave digestion system

with built in, non-contact temperature and pressure measurement – type

Speedwave MWS – 3 (Berghof), with microwave power 1 450 W. At the

completion of this process, samples were diluted to 100 ml with deionized

water.

Trace metals concentrations were determined by graphite furnace

atomic absorption spectrometry (GF-AAS), using an SOLAAR M6 DUAL

Zeeman, Thermo Electron – UNICAM equipment.

RESULTS AND DISCUSSION

Data obtained for abiotic elements revealed a highly variability

especially for organic pollutant.

In water samples, heavy metals had concentrations between 9.98 μg/L

and 20.14 μg/L for Cu; 0.98 μg/L and 2.15 μg/L for Cd; 15.21 μg/L and 29.31

μg/L for Pb; 5.79 μg/L and 9.25 μg/Lfor Cr and 2.15 μg/L and 4.74 μg/L for

Ni. The highest levels were measured for Pb (24.72 g/L in fluvial water -

Sulina arm; 25.34 g/L in Musura Bay and 29.31 g/L in marine waters -

Sfântul Gheorghe arm discharging area S (Fig. 2).

In sediments heavy metals had concentrations between 2.03 μg/g dry

sediment and 20.14 μg/g dry sediment for Cu; 0.30 μg/g dry sediment and 2.95

μg/g dry sediment for Cd; 4.11 μg/g dry sediment and 36.96 μg/g dry sediment

for Pb; 5.32 μg/g dry sediment and 18.46 μg/g dry sediment for Cr; 2.81 μg/g

dry sediment and 46.66 μg/g dry sediment for Ni and 1.08 μg/g dry sediment

and 9.24 μg/g dry sediment for Ni. The highest levels were measured for Ni

(46.6 g/ g dry sediment in fluvial area – Sfântul Gheorghe arm; 37.87 g/g

dry sediment in Musura Bay), Cu (20.14 μg/g dry sediment in fluvial area –

Sfântul Gheorghe arm) and Pb (37.87 g/g dry sediment in fluvial area –

Sulina arm; 36.96 μg/g dry sediment in marine waters - Sfântul Gheorghe arm

discharging area) (Fig. 3).

0 10 20 30 40 50 60 70

Chilia Arm

Sulina Arm - Tulcea

Sulina Arm - Crisan

Sulina Arm - Sulina

Sf. Gheorghe Arm- Murighiol

Musura Bay - N

Musura Bay - V

Musura Bay- S

Musura Bay - E

Sulina discharge area- S

Sf. Gheorghe discharge area - N

Sf. Gheorghe discharge area- S

µg/l

Cu

Cd

Pb

Cr

Ni

Fig. 2 - Concentration of heavy metals in water samples collected from Danube

mouth and Black Sea area influenced by Danube

0 20 40 60 80 100 120

Chilia Arm

Sulina Arm - Tulcea

Sulina Arm- Sulina

Sf. Gheorghe Arm - Murighiol

Musura Bay - V

Musura Bay- S

Musura Bay- E

Sulina discharge area - S

Sf. Gheorghedischarge area - N

Sf. Gheorghe discharge area - S

µg/g d.w .

Cu

Cd

Pb

Cr

Ni

Li

Fig. 3 - Concentration of heavy metals in sediment samples collected from Danube

mouth and Black Sea area influenced by Danube

Polycyclic aromatic hydrocarbons were detected in all water samples

in concentrations between 0.11 and 1969.41 ng/L. The highest level of PAHs

was measured in southern part of Musura Bay (1520.11 ng/L Naphtalene,

1886.57 ng/L Phenanthrene, 1969.41 ng/L Anthracene and 1433.78 ng/L

Indeno(1,2,3-c,d)pyrene), and also in Sfântul Gheorghe arm discharging area

(1057.23 ng/L Naphtalene, 1542.39 ng/L Phenanthrene, 1610.17 ng/L

Anthracene) (Fig. 4).

Fig. 4 - Concentration of polycyclic aromatic hydrocarbons in water samples

collected from Danube mouth and Black Sea area influenced by Danube

In sediments polycyclic aromatic hydrocarbons varied between 0,02

and 186.14 ng/g dry sediment. The highest values were measured in fluvial

area at Murighiol (109.8 ng/g dry sediment Nphthalene and 182.92 ng/g dry

sediment Phenanthrene); in V Musura Bay (186.14 ng/g dry sediment

Naphtalene and 327.71 ng/g dry sediment Anthracene in Sfântul Gheorghe

arm discharging area northen part (362.46 ng/g dry sediment Phenanthrene)

(Fig. 5).

Organochlorine pesticides (HCB, lindane, heptachlor, aldrin, dieldrin,

endrin, DDE, DDD, DDT) had concentrations between 1.7 ng/L and 586.73

ng/L in water and 1.7 ng/g dry sediment and 339.21 ng/g dry sediment.

Chilia

arm

Sulina

Arm

-Tulc

ea

Sulina

Arm

-Cris

an

Sulina

Arm

-Sul

ina

Sf. Ghe

orgh

e Arm

-Mur

ighio

l

Mus

ura

Bay-N

Mus

ura

Bay-V

Mus

ura

Bay-S

Mus

ura

Bay-E

Sulina

disc

harg

e ar

ea-S

Sf. Ghe

orgh

e dis

char

ge a

rea-

N

Sf.Ghe

orgh

e dis

char

ge a

rea-

S

0

200

400

600

800

1000

1200

1400

1600

1800

2000

ng/L

Naphthalene

Acenaphthylene

Acenaphthene

Fluorene

Phenanthrene

Anthracene

Floranthene

Pyrene

Benzo[a]antracene

Crysene

benzo(k)fluoranthene

Benzo[a]pyrene

Benzo(ghi)perilen

Dibenzo[a,h]anthracene

Indeno(1,2,3-c,d)piren

Fig. 5 - Concentration of polycyclic aromatic hydrocarbons in sediment samples collected

from Danube mouth and Black Sea area influenced by Danube

The highest levels were measured for HCB, lindane and aldrin both in

water and sediments (Fig. 6 and Fig. 7). In water maximum concentrations

were measured for HCB in fluvial – Sulina arm (334.63 ng/L at Crisan site and

360.14 ng/L at Tulcea site); for lindane în marine area E Musura Bay (586.73

ng/L) and in Sulina arm discharging area (488.59) ng/L and aldrin in marine

area (112.16 ng/L - 543.61 ng/L). In sediments maximum concentrations were

measured in fluvial area on Sfantul Gheorghe arm at Murighiol site (125.22

ng/g dry sediment HCB, 124.33 ng/g dry sediment lindane and 339.21 ng/g

dry sediment aldrin) and in marine discharging area for Sfântul Gheorghe arm

(88.21 ng/g dry sediment HCB, 144.08 ng/g dry sediment lindane and 188.66

ng/g dry sediment aldrin).

Chili

a A

rm

Sulin

a A

rm-T

ulc

ea

Sulin

a A

rm-S

ulin

a

Sf. G

heorg

he A

rm-M

urighio

lM

usu

ra B

ay-

VM

usu

ra B

ay-

SM

usu

ra B

ay-

E

Sulin

a d

isch

arg

e a

rea-S

Sf. G

heorg

he d

isch

arg

e a

rea-N

Sf.G

heorg

he d

isch

arg

e a

rea-S

0

50

100

150

200

250

300

350

400

ng

/g d

.s.

Naphthalene

Acenaphthylene

Acenaphthene

Fluorene

Phenanthrene

Anthracene

Floranthene

Pyrene

Benzo[a]antracene

Crisen

benzo(k)fluoranthene

Benzo[a]pyrene

Benzo(ghi)perilen

Dibenzo[a,h]anthracene

Indeno(1,2,3-c,d)piren

Fig. 6 - Concentration of organochlorine pesticides in water samples collected from Danube

mouth and Black Sea area influenced by Danube

Fig. 7 - Concentration of organochlorine pesticides in sediment samples collected from

Danube mouth and Black Sea area influenced by Danube

According to Romanian legislation (Order 161/2006, Dec.351/2005)

there are some values that exceeded the quality standards both in water and

sediments. These values refer to lead, HCB, lindane, aldrin, phenanthrene,

anthracene, benzo[a]pyrene and benzo(ghi)perilene in water and cadmium,

naphthalene, phenanthrene, and antracen in sediments.

As for biota heavy metals concentrations varied between 0,01 μg/g

tissue and 28.89 μg/g tissue in mollusks whole body. The highest levels were

measured for copper (21.95 μg/g tissue for Rapana sampled in Portiţa site and

28.89 μg/g tissue for Rapana sampled in Zaton site). The others metals had

concentrations less than 4 μg/g tissue (Fig. 8). In fish muscle heavy metals

concentrations varied between 0.03 μg/g tissue and 2.19 μg/g tissue. The

highest concentrations were measured for copper and chromium (Fig. 9).

0 5 10 15 20 25 30 35

Mytilus - Sulina 30m

Mytilus-Mila9 30m

Mytilus-Portita 10m

Mytilus-Zaton 20m

Scapharca - Sulina 10m

Mya- Sf. Ghe. 30m

Mya - Perisor 20m

Rapana - Portita 10 m

Rapana - Zaton 10 m

ug/g tissue

Cu Cd Pb Ni Cr

Fig. 8 - Concentration of heavy metals in mollusk tissue collected from Black Sea

area influenced by Danube

Organochlorine pesticides found in biota were HCB, lindane,

heptachlor, aldrin, DDE, DDD and DDT. Their concentrations varied between

0.02 μg/g tissue and 2.42 μg/g tissue in mollusks whole body and 0.009 μg/g

tissue and 0.944 μg/g tissue in fish. The highest levels were measured in

mollusks (> 1 μg/g tissue for lindane, heptachlor, DDD and DDT) (Fig. 10). In

fish the biggest concentrations were measured for DDT: between 0.42 μg/g

tissue and 0.944 μg/g tissue, especially for marine species (Fig. 11).

0 2 4 6 8 10 12

Exos lucius (Sulina)

Cyprinus carpio (Sulina)

Carassius auratus (Sulina)

Abramis brama danubii (Sulina)

Silurus glanis(Sulina)

Perca fluviatilis (Sf.Ghe.)

Rutilus pigus (Sf. Ghe.)

Scardinius erytrophtalmus (Sf. Ghe.)

Salmo trutta labrax (Sulina)

Trachurus mediterraneus ponticus (Sf. Ghe.)

Neogobius melonostomus (Sf. Ghe.)

Sprattus sprattus (Sf. Ghe.)

Alosa caspia nordmanni (Sf. Ghe.)

Engraulis encrasicolus(Sf. Ghe.)

Merlangius merlangus euxinus(Sf. Ghe.)

ug/g tissueCu Cd Pb Ni Cr

Fig. 9 - Concentration of heavy metals in fish tissue collected from Danube mouth

and Black Sea area influenced by Danube

Fig. 10 - Concentration of organochlorine pesticides in mollusk tissue collected

from Black Sea area influenced by Danube

0 1 2 3 4 5 6 7

Mytilus - Sulina 30m

Mytilus-Mila9 30m

Mytilus-Portita 10m

Mytilus-Zaton 20m

Scapharca - Sulina 10m

Mya- Sf. Ghe. 30m

Mya - Perisor 20m

Rapana - Portita 10 m

Rapana - Zaton 10 m

ug/tesut

HCB Lindan Heptaclor Aldrin Dieldrin Endrin DDE DDD DDT

Fig. 11 - Concentration of organochlorine pesticides in fish tissue collected from

Danube mouth and Black Sea area influenced by Danube

Fig. 12 - Concentration of polycyclic aromatic hydrocarbons in fish tissue collected from

Danube mouth and Black Sea area influenced by Danube

0 0,2 0,4 0,6 0,8 1 1,2 1,4 1,6 1,8 2

Exos lucius (Sulina)

Cyprinus carpio (Sulina)

Carassius auratus (Sulina)

Abramis brama danubii (Sulina)

Silurus glanis (Sulina)

Perca fluviatilis (Sf.Ghe.)

Rutilus pigus (Sf. Ghe.)

Scardinius erytrophtalmus (Sf. Ghe.)

Salmo trutta labrax (Sulina)

Trachurus mediterraneus ponticus (Sf. Ghe.)

Neogobius melonostomus (Sf. Ghe.)

Sprattus sprattus (Sf. Ghe.)

Alosa caspia nordmanni (Sf. Ghe.)

Engraulis encrasicolus(Sf. Ghe.)

Merlangius merlangus euxinus(Sf. Ghe.)

ug/g tissue

HCB Lindane Heptachlor Aldrin Dieldrin Endrin p,p'DDE p,p'DDD p,p'DDT

0 0,2 0,4 0,6 0,8 1 1,2

Exos lucius (Sulina)

Cyprinus carpio (Sulina)

Carassius auratus (Sulina)

Abramis brama danubii (Sulina)

Silurus glanis (Sulina)

Perca fluviatilis (Sf.Ghe.)

Rutilus pigus (Sf. Ghe.)

Scardinius erytrophtalmus (Sf. Ghe.)

Salmo trutta labrax (Sulina)

Trachurus mediterraneus ponticus (Sf. Ghe.)

Neogobius melonostomus (Sf. Ghe.)

Sprattus sprattus (Sf. Ghe.)

Alosa caspia nordmanni (Sf. Ghe.)

Engraulis encrasicolus(Sf. Ghe.)

Merlangius merlangus euxinus(Sf. Ghe.)

ug/g tissue

Naphthalene Acenaphthene Acenaphthylene Fluorene Phenanthrene

Floranthene Pyrene Crysene Anthracene Benzo[a]antracene

Fig. 13 - Concentration of polycyclic aromatic hydrocarbons in mollusk tissue collected from

Danube mouth and Black Sea area influenced by Danube

According to Romanian legislation (Order 97/7 Oct.2005; Order 23/1

Febr.2007, Normes/16 Dec.1998) there are some values that exceeded the

quality standards for biota. These values refer to lead in some marine fish

species, cadmium in many fresh water fish species, lindane, heptachlor and

DDT in molluscs whole body sampled from Mila 9 30m, Portiţa 10m and

Sfântul Gheorghe 30m sites, endrin in some fresh water fish species and DDT

in some marine fish species.

CONCLUSIONS

1. The distribution of heavy metals, organochlorine pesticides and

polycyclic aromatic hydrocarbons in abiotic elements underlined the

accumulation of pollutants in eastern and southern part of Musura Bay and in

discharging marin area of Sulina and Sfântul Gheorghe arms.

2. The bioaccumulation of heavy metals and organochlorine

pesticides is higher in mollusks than fish tissue, most probably because of

biotope mollusk’s “fidelity”.

0 0,1 0,2 0,3 0,4 0,5 0,6 0,7 0,8 0,9 1

Mytilus-Portita 10m

Mytilus-Zaton 20m

Scapharca - Sulina 10m

Mya- Sf. Ghe. 30m

Mya - Perisor 20m

Rapana - Portita 10 m

ug/g tissueNaphthalene Acenaphthene Acenaphthylene Fluorene Phenanthrene

Floranthene Pyrene Crysene Anthracene Benzo[a]antracene

3. The bioaccumulation of polycyclic aromatic hydrocarbons is

less obviously. Their concentration in tissue is smaller than for the others

pollutants and the levels are similar in mollusks and fish.

BIBLIOGRAPHY :

***, 1998 - Norma igienico - sanitarǎ din 16 decembrie 1998 pentru alimente.

***, 2005 - Hotararea nr. 351/2005 privind aprobarea Programului de

eliminare treptata a evacuarilor, emisiilor şi pierderilor de substanţe

prioritar periculoase.

***, 2005 - Ordinul nr. 97 din 7 octombrie 2005 privind aprobarea Normei

sanitare veterinare şi pentru siguranţa alimentelor privind anumiţi

contaminanţi din alimentele de origine animală şi nonanimală.

***, 2006 - Ordinul ministrului mediului şi gospodǎririi apelor nr. 161/2006

pentru aprobarea Normativului privind clasificarea calitatii apelor de

suprafaţǎ în vederea stabilirii stǎrii ecologice a corpurilor de apǎ.

***, 2007 - Ordinul nr.23 din 1 februarie 2007 privind aprobarea Normei

sanitare veterinare şi pentru siguranţa alimentelor privind stabilirea

limitelor maxime de reziduuri de pesticide din conţinutul sau de pe

suprafaţa produselor alimentare de origine animală.