hydrocarbon contamination in mussels from guanabara bay

3
Hydrocarbon contamination in mussels from Guanabara Bay Luı ´s A. Azevedo, Ina ´i M.R. de Andrade Bru ¨ ning, Isabel Moreira * Department of Chemistry, Pontifı ´cia Universidade Cato ´ lica, Rua Marque ˆs de S.Vicente, 225, 22453-900 Ga ´ vea, Rio de Janeiro, Brazil Guanabara Bay is the most important embayment on the south eastern coast of Brazil. Three cities, Rio de Ja- neiro, Nitero ´i and Sa ˜o Gonc ¸alo are located in the sur- roundings with an estimated total population of 11 million. Within the state of Rio de Janeiro, most of the petroleum activities in Brazil are concentrated, and around Guanabara Bay two refineries, several petro- chemical plants, oil terminals, shipyards and offshore platform maintenance installations render the bay prone to hydrocarbon contamination. Several studies have examined hydrocarbon contam- ination in the waters (Hamacher, 1996; Hamacher et al., 2000) and sediments (Hamacher, 1996; Lima, 1996) of Guanabara Bay. The present work searched for ali- phatic and polyaromatic hydrocarbon (PAH) contami- nation in mussels Perna perna, that grow near the entrance of Guanabara Bay. Previously, such mussels have been investigated for organochlorine accumulation (Xavier de Brito et al., 2002). Guanabara Bay is a typical tropical estuary with warm, wet summers and dry, cool winters. A detailed description of the Bay has been presented before (Kjerfve et al., 1997; Godoy et al., 1998; Xavier de Brito et al., 2002). Fig. 1 presents the map of Guanabara Bay and the location of the five sampling stations. These sta- tions were chosen in close vicinity to potential pollution sources and in areas where mussel fishing or cultivation is carried out for human consumption. Station 1 is located in the mussel cultivation area of Jurujuba Fisherman Corporation; station 2, between the Santos Dumont City Airport and the Marina da Glo ´ ria, which is a public mussel collection place; station 3 is located around the second biggest pillar on the left side of the Rio de Janeiro-Niteroi bridge; station 4 is on the second biggest pillar of the right side of the same bridge; and station 5 lies in Niteroi, on the Boa Viagem Beach, and is also a public mussel collection area. In the innermost area of the bay, where mussels are seldom found, no samples were collected. The mussels were manually collected in August (winter) and in December (summer), 1996. In December, no specimen was col- lected in station 4, because suitably sized mussels for monitoring could not be found, due to intense summer fishing activity. The collected mussels were wrapped in aluminum foil and kept in iceboxes until reaching the laboratory. They were then selected according to size (4–6 cm), with 10 individuals composing each sample. The soft tissues were separated from the valves and were freeze-dried. The material was ground and homogenized and stored at 10 °C until extraction. The analytical procedure used to detect the aromatic and aliphatic contamination was that recommended by the National Bureau of Standards (Wise et al., 1980). Freeze-dried tissue (2 g) was Sohxlet extracted for 20 h using 200 ml of methanol; saponification was car- ried out by adding 50 ml of KOH 0.5 N aqueous solu- tion, and the sample was then extracted for a further 4 h. The methanol extracts were shaken three times in a separation funnel with 50 ml of normal hexane. The final hexane extracts were passed through an anhydrous sodium sulphate column and finally concentrated in a rotary evaporator to approximately 1 ml. Octadecene-1 and 9,10 dihydroanthracene were added to the extracts to evaluate the extraction recoveries for the aliphatic and aromatic fractions, respectively. The hydrocarbon extracts were separated into aliphatic and aromatic fractions in a silica:alumina (1:1) column, using 25 ml of n-hexane for eluting the aliphatic hydrocarbons, 25 ml of hexane:dichloromethane mixture (4:1) and 25 ml of hexane:dichloromethane (1:1) for recovering the aromatics. On the top of the column, a thin layer of anhydrous sodium sulphate was added in order to eliminate eventual water residues. The separation be- tween the aliphatic and aromatic fractions was moni- tored by UV spectrometry. Both fractions were analyzed in a gas chromatograph (Hewlett Packard 6890) equipped with a hydrogen flame ionization detector and a fused silica DB-5 capillary col- umn of 30 m · 0.25 mm dimensions and film thickness of 0.25 lm. Splitless mode injections of 1 ll volume were used for all samples. Column temperature programming started from 50 °C to 280 °C at a rate of 5 °C min 1 , with a final isothermal period of 15 min. Helium and nitrogen were carrier and make up gases at the flow rates of 1.0 ml min 1 , and 44.0 ml min 1 , respectively. Injector and detector temperatures were 260 °C and 300 °C, respectively. Quantitation was carried out by comparison with internal standards, naphthalene and normal hexade- cane, respectively for aliphatic and aromatic fractions. A certified mussel sample (IAEA 142) was analyzed according to this procedure; the results were within the confidence intervals for both aliphatic and polyaromatic hydrocarbons. The average extraction recovery for the * Corresponding author. Fax: +55 21 3114 1637. E-mail address: [email protected] (I. Moreira). 1120 Baseline / Marine Pollution Bulletin 49 (2004) 1109–1126

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Hydrocarbon contamination in mussels from Guanabara Bay

Luıs A. Azevedo, Inai M.R. de Andrade Bruning, Isabel Moreira *

Department of Chemistry, Pontifıcia Universidade Catolica, Rua Marques de S.Vicente, 225, 22453-900 Gavea, Rio de Janeiro, Brazil

Guanabara Bay is the most important embayment on

the south eastern coast of Brazil. Three cities, Rio de Ja-

neiro, Niteroi and Sao Goncalo are located in the sur-

roundings with an estimated total population of 11million. Within the state of Rio de Janeiro, most of

the petroleum activities in Brazil are concentrated, and

around Guanabara Bay two refineries, several petro-

chemical plants, oil terminals, shipyards and offshore

platform maintenance installations render the bay prone

to hydrocarbon contamination.

Several studies have examined hydrocarbon contam-

ination in the waters (Hamacher, 1996; Hamacher et al.,2000) and sediments (Hamacher, 1996; Lima, 1996) of

Guanabara Bay. The present work searched for ali-

phatic and polyaromatic hydrocarbon (PAH) contami-

nation in mussels Perna perna, that grow near the

entrance of Guanabara Bay. Previously, such mussels

have been investigated for organochlorine accumulation

(Xavier de Brito et al., 2002).

Guanabara Bay is a typical tropical estuary withwarm, wet summers and dry, cool winters. A detailed

description of the Bay has been presented before

(Kjerfve et al., 1997; Godoy et al., 1998; Xavier de Brito

et al., 2002). Fig. 1 presents the map of Guanabara Bay

and the location of the five sampling stations. These sta-

tions were chosen in close vicinity to potential pollution

sources and in areas where mussel fishing or cultivation

is carried out for human consumption.Station 1 is located in the mussel cultivation area of

Jurujuba Fisherman Corporation; station 2, between

the Santos Dumont City Airport and the Marina da

Gloria, which is a public mussel collection place; station

3 is located around the second biggest pillar on the left

side of the Rio de Janeiro-Niteroi bridge; station 4 is

on the second biggest pillar of the right side of the same

bridge; and station 5 lies in Niteroi, on the Boa ViagemBeach, and is also a public mussel collection area. In the

innermost area of the bay, where mussels are seldom

found, no samples were collected. The mussels were

manually collected in August (winter) and in December

(summer), 1996. In December, no specimen was col-

lected in station 4, because suitably sized mussels for

monitoring could not be found, due to intense summer

fishing activity. The collected mussels were wrapped inaluminum foil and kept in iceboxes until reaching the

laboratory. They were then selected according to size

(4–6cm), with 10 individuals composing each sample.

The soft tissues were separated from the valves and were

freeze-dried. The material was ground and homogenizedand stored at �10 �C until extraction. The analytical

procedure used to detect the aromatic and aliphatic

contamination was that recommended by the National

Bureau of Standards (Wise et al., 1980).

Freeze-dried tissue (2g) was Sohxlet extracted for

20h using 200ml of methanol; saponification was car-

ried out by adding 50ml of KOH 0.5N aqueous solu-

tion, and the sample was then extracted for a further4h. The methanol extracts were shaken three times in

a separation funnel with 50ml of normal hexane. The

final hexane extracts were passed through an anhydrous

sodium sulphate column and finally concentrated in a

rotary evaporator to approximately 1ml. Octadecene-1

and 9,10 dihydroanthracene were added to the extracts

to evaluate the extraction recoveries for the aliphatic

and aromatic fractions, respectively. The hydrocarbonextracts were separated into aliphatic and aromatic

fractions in a silica:alumina (1:1) column, using 25ml

of n-hexane for eluting the aliphatic hydrocarbons,

25ml of hexane:dichloromethane mixture (4:1) and

25ml of hexane:dichloromethane (1:1) for recovering

the aromatics. On the top of the column, a thin layer

of anhydrous sodium sulphate was added in order to

eliminate eventual water residues. The separation be-tween the aliphatic and aromatic fractions was moni-

tored by UV spectrometry.

Both fractions were analyzed in a gas chromatograph

(Hewlett Packard 6890) equipped with a hydrogen flame

ionization detector and a fused silica DB-5 capillary col-

umn of 30m · 0.25mm dimensions and film thickness of

0.25lm. Splitless mode injections of 1ll volume were

used for all samples. Column temperature programmingstarted from 50 �C to 280 �C at a rate of 5 �Cmin�1, with

a final isothermal period of 15min. Helium and nitrogen

were carrier and make up gases at the flow rates of

1.0mlmin�1, and 44.0mlmin�1, respectively. Injector

and detector temperatures were 260 �C and 300 �C,respectively.

Quantitation was carried out by comparison with

internal standards, naphthalene and normal hexade-cane, respectively for aliphatic and aromatic fractions.

A certified mussel sample (IAEA 142) was analyzed

according to this procedure; the results were within the

confidence intervals for both aliphatic and polyaromatic

hydrocarbons. The average extraction recovery for the

* Corresponding author. Fax: +55 21 3114 1637.

E-mail address: [email protected] (I. Moreira).

1120 Baseline / Marine Pollution Bulletin 49 (2004) 1109–1126

aliphatic fractions was 92.8 ± 4.0%. For the aromatic

fractions, a mean value of 90.4 ± 2.9% was found.Fig. 2 shows the distribution of the total aliphatic

hydrocarbon concentrations for the collected samples

in winter and summer seasons. The concentrations

(C10–C32) were higher in August (winter) and ranged

from 520 to 1461ngg�1 dry weight, whereas in Decem-

ber (summer) the range was 309 to 829ngg�1 dry

weight. The most contaminated mussels were those col-

lected in winter in stations 3 and 4 located on the pillarsof the bridge. Those mussels filter water coming from

the inner area of the Bay, where refineries and petroleum

terminals are located.

The gas chromatographic profiles of the aliphatic

fraction from station 2 samples (both August and

December) exhibited higher concentration of naphth-

enic compounds in the C26 region, which is typical of

lube oils. The evaluation of the aliphatic fraction chro-matograms shows a similar pattern for each station for

both summer and winter seasons. This similarity leads

to the conclusion that the qualitative contamination

sources remain constant, although the higher concentra-

tions in the dry season are derived from the lower water

input to the Bay. When compared to other inhabited

coastal areas (Anderlini et al., 1981; Law and And-

rulewicz, 1983; Martin and Castle, 1984; Shaw et al.,1986) the aliphatic concentrations of Guanabara Bay

mussels are among the lowest.

Among the polyaromatic hydrocarbons, fluoranthene

was predominant at all stations, and its range was

approximately constant in winter and summer seasons

(41.5–128.9ngg�1 dry weight in August and 59.7–

123.5ngg�1 dry weight in December). Other coastal sys-

tems have reported this polyaromatic as the predomi-nant pollutant in mussels (Shaw et al., 1986;

Shchekaturina et al., 1995). Also for the aromatic frac-

tions, a similar distribution for both seasons in each

location was observed, indicating constant qualitative

exposure throughout the year. Total polyaromatic pol-

lution (Fig. 3), was also considerably higher in winter

(173–432ngg�1 dry weight) than in summer (68–

375ngg�1 dry weight). Again station 2 presented higherpolyaromatic concentrations for both seasons.

The higher polyaromatic hydrocarbon contamination

of station 2, located near the entrance channel of the

largest marina of the Bay, indicates that intense sport

and leisure boat traffic and maintenance contribute

significantly to PAH pollution in the Bay. Station 4,

however, presented the second highest concentration of

aliphatic and polyaromatic hydrocarbons, with achromatographic pattern of the aliphatic fraction typical

Fig. 1. Guanabara Bay with sampling collection sites.

Fig. 2. Total aliphatic hydrocarbon concentrations (ng/g dry weight).

Fig. 3. Total PAH concentrations (ng/g dry weight).

Baseline / Marine Pollution Bulletin 49 (2004) 1109–1126 1121

of petroleum origin, with marked UCM (unresolved

complex mixture) proving the industrial origin of the

contamination. The polyaromatic qualitative distribu-

tion is similar to those of other stations. In comparison

with other studies (Cocchieri et al., 1990; Shaw et al.,

1986; Shchekaturina et al., 1995) Guanabara Bay pre-sented mussel polyaromatic concentrations within the re-

ported ranges.

Acknowledgments

This work was sponsored by CAPES. Luis A. Azev-

edo and Inai M.R. de Andrade Bruning are thankfulrespectively to CNPq and FAPERJ for financial support.

References

Anderlini, V.C., Al-Harmi, L., De Lappe, B.W., Risebrough, R.W.,

Walker, W., Simoneit, B.R.T., Newton, A.S., 1981. Distribution of

hydrocarbons in the oyster, Pinctada margaratifera, along the coast

of Kuwait. Marine Pollution Bulletin 12, 57–62.

Cocchieri, R.A., Arnese, A., Minicucci, A.M., 1990. Polycyclic

aromatic hydrocarbons in marine organisms from Italian central

Mediterranean coasts. Marine Pollution Bulletin 21, 15–18.

Godoy, J.M., Moreira, I., Braganca, M.J., Wanderley, C., Mendes,

L.B., 1998. A study of Guanabara Bay sedimentation rates.

Journal of Radioanalytical and Nuclear Chemistry 227, 157–160.

Hamacher, C., 1996. Determinacao de hidrocarbonetos em amostras

de agua e sedimento da Baıa de Guanabara. Master Thesis,

Pontifıcia Universidade Catolica, Rio de Janeiro, Brazil, 101pp.

Hamacher, C., Xavier de Brito, A.P., De Andrade Bruning, I.M.R.,

Wagener, A., Moreira, I., 2000. The determination of PAH by UV-

fluorescence spectroscopy in water of Guanabara Bay, Rio de

Janeiro, Brazil. Revista Brasileira de Oceanografia 48, 167–

170.

Kjerfve, B., Ribeiro, C.H.A., Dias, G.T.H., Fillipo, A.M., Quaresma,

V.S., 1997. Oceanographic characteristics of an impacted coastal

bay: Baıa de Guanabara, Rio de Janeiro, Brazil. Continental Shelf

Research 17, 1609–1643.

Law, R., Andrulewicz, E., 1983. Hydrocarbons in water, sediment and

mussel from Southern Baltic Sea. Marine Pollution Bulletin 14,

289–293.

Lima, A.L.C., 1996. Geocronologia de hidrocarbonetos poliaroma-

ticos (PAHs). Estude de caso: Baıa de Guanabara. Master

Thesis, Pontifıcia Univercidade Catolica, Rio de Janeiro, Brazil,

106pp.

Martin, M., Castle, W., 1984. Petrowatch: petroleum hydrocarbons,

synthetic organic compounds, and heavy metals in mussels from

the Monterey Bay area of Central California. Marine Pollution

Bulletin 15, 259–266.

Shaw, D.G., Hogan, T.E., McIntosh, D.I., 1986. Hydrocarbons in

bivalve molluscs of Port Valdez, Alaska: consequences of five years�permitted discharge. Estuarine, Coastal and Shelf Science 23, 863–

872.

Shchekaturina, T.L., Khesina, A.L., Mironov, O.G., Krivosheeva,

L.G., 1995. Carcinogenic polycyclic aromatic hydrocarbons in

mussel from the Black Sea. Marine Pollution Bulletin 30, 38–

40.

Wise, S.A., Chesler, S.N., Guenther, H.S., Hertz, H.S., Hilpert, L.R.,

May, W.E., Parris, R.M., 1980. Interlaboratory comparison of

determinations of trace level hydrocarbons in mussels. Analytical

Chemistry 52, 1828–1833.

Xavier de Brito, A.P., De Andrade Bruning, I.M.R., Moreira, I., 2002.

Chlorinated pesticides in mussels from Guanabara Bay, Rio de

Janeiro, Brazil. Marine Pollution Bulletin 44, 79–81.

0025-326X/$ - see front matter � 2004 Elsevier Ltd. All rights reserved.

doi:10.1016/j.marpolbul.2004.10.003

Intersex in Roe�s abalone (Haliotis roei) in Western Australia

Sara Sloan, Marthe Monique Gagnon *

Department of Environmental Biology, Curtin University of Technology, Perth 6845, Australia

Since the 1970s, tributyltin (TBT) has been used as

the active ingredient in antifouling paints applied tothe hulls of vessels to prevent the attachment and

growth of fouling organisms. The leaching of TBT in

marine waters has been associated with the induction

of imposex, the imposition of male characters onto a fe-

male organism. To date, imposex has been reported in

63 genera and 140 species worldwide (Terlizzi et al.,

2004; Reitsema et al., 2002). Even at extremely low con-

centrations such as 0.5ngL�1 imposex has been ob-

served in Nucella lapillus, a marine snail (Bryan et al.,

1987). The deleterious effects observed globally triggeredthe adoption of legislative restrictions regarding the

application of TBT in many countries. In 1991, the

Western Australian government implemented legislation

restricting the use of TBT to vessels greater than 25m in

length.

In 1998–1999, Reitsema et al. (2003) surveyed 16 sites

in the Perth metropolitan area, and found a significant

reduction in imposex symptoms in the whelk Thais

orbita at 11 sites relative to a 1991 survey. However,

sites close to commercial harbours and docks still main-

tained 100% imposexed females with 30% of these with

permanent sterility. Reitsema et al. (2003) also observed

* Corresponding author. Tel.: +61 8 92663723; fax: +61 8 92662495.

E-mail address: [email protected] (M.M. Gagnon).

1122 Baseline / Marine Pollution Bulletin 49 (2004) 1109–1126