The Science of the Total Environment, 125 (1992) 359-372 Elsevier Science Publishers B.V., Amsterdam

359

Relationships among the species of copper, organic compounds and bioaccumulation along the

mariculture area in Taiwan

T.C.Hung a,b and B.C.Han c

alnstitute of Chemistry, Academia Sinica, Taipei 11529, Taiwan, ROC blnstitute of Oceanography, National Taiwan University, Taipei 10764 Taiwan, ROC

CDepartment of Public Health, Taipei Medical College, TaipeL Taiwan, ROC

ABSTRACT

In Taiwan the first incident of green discoloration in the oysters Crassostrea gigas was observed in the Charting coastal area in January 1986 and the mortality reports appeared 3 months later. The cause of green oysters was identified as copper pollution. The copper con- tent of the oysters was extremely high (2100/zg/g and 4400 #g/g of dry weight in January 1986 and January 1989, respectively). In this paper some recent data on distribution of copper species (complexed by inorganic and organic ligands, labile and non-labile, polar and non- polar) and forms (dissolved and particulate), dissolved organic compounds (such as lipids, acid- and base-mobile organic compounds) and copper assimilative capacity in sea water are reported. In general, high concentrations of particulate copper (range from 3.09 to 732/~g/i) and non-labile organic copper (over 70% of total dissolved copper) were observed. Relatively low values of polar organic copper (less than about 48% of the total organic copper) indicated that non-polar organic complexes from man-made organic pollutants were the major com- plexes in the study area.

Key words: copper species; copper assimilative capacity; dissolved organic compounds; copper bioaccumulation in oysters; ecological parameters

INTRODUCTION

One of the important oyster mariculture areas in Taiwan is along the Charting coastal region. Recently, due to the dense population and heavy in- dustry expansion, domestic and industrial wastes, mostly without any treat- ment, have been discharged through the Erhjin Chi River into the Charting coastal area. The first incident of green discoloration in oysters (Crassostrea gigas) in the Charting coastal area was observed in January 1986 and mor- tality reports appeared 3 months later. The major factor to cause the green

0048-9697/92/$05.00 © 1992 Elsevier Science Publishers B.V. All rights reserved

360 T.C. HUNG AND B.C. HAN

oysters was immediately identified as being copper pollution by local copper recycling operations [1]. Hung [2] also indicated that high adenylate energy charge (AEC) value (0.84) was obtained in the oysters during the onset of discoloration and the low AEC value (0.21) was observed in their mortality period. Recently, we found that as much as 98% of dissolved copper can be involved in organic complexes collected from Chung-Kang Chi coastal area [3]. Chen et al. [4] also reported that the concentrations of organic copper ranged from 0.21 to 4.54/zg/1 (approx. 9.20-87.6% of total dissolved copper concentrations) in the western coastal water; these were much higher than those ranging from 0.16 to 0.46/zg/1 (approx. 5.50-14.0% of total dissolved

x , , ,~ . : . "~ 23"00" %/~ A n - p i n

H o r b o r 0 I 2 3 /- 5

Km

©11 06

22"55' 010 s015

09

s

22"50"

05" 120°10 " 14'

Fig. 1. Sampling stations along the Charting coastal area (S: Oyster cultural stations).

COPPER POLLUTION IN THE MARICULTURE AREA OF TAIWAN 361

copper) off the eastern coast of Taiwan. The purpose of this paper is to con- tinue our observations on the causes of green oysters along the Charting coastal area. The focuses are made on the relationships among the copper species (inorganic and organic, labile and non-labile, polar and non-polar) and forms (particulate and dissolved), dissolved organic compounds (such as lipids, acid-mobile and base-mobile compounds), copper assimilative capa- city, and the processes of bioaccumulation and elimination of copper in oysters.

EXPERIMENTAL METHODS

Water samples and oysters were collected from the Charting coastal area including the Erhjin River estuary during the period from July 1988 to March 1989. The sampling locations are shown in Fig. 1. Water samples at a depth of 3 m were collected with metal-free Van Dorn bottles on board either the research vessel Ocean Researcher 1 or fishing boats. Immediately after the collection, water samples, filtered through 0.45 #m glass fiber filters, were analyzed for hydrographical, chemical and ecological parameters. The protocol for analysis of these parameters is shown in Fig. 2. The methods for particulate organic carbon (POC), dissolved organic carbon (DOC), chlorophyll a, adenosine triphosphate (ATP), nitrate, phosphate and copper complexing capacity (CC) analyses were described previously by Hung et al. and Chen et al. [1,4-5]. The analysis of the species (inorganic and organic, polar and non-polar, labile and non-labile) and the forms (dissolved and par- ticulate) of copper were preformed by the differential pulse anodic stripping voltametric (DPASV) and the C-18 bound Sep-Pak cartridge methods [4].

Seawater Filtered I Unfiltered

I I DissOlved Cu f r a c t i o n complexometr ic

Particulate I titration fraction C-18 ~ep-pak NatUral pH pll~B DOC ~u

unretained retained DPASV tit ation Comp}exing

I I [ I CH30H Labile Cu Total capac i ty & Eluate dis,Cu Complexing stability (Labile+Nonlabile CU) capacity & constants AIP Chl.a Cu(p} POC pH<2 stability

i 0PAsv 1 1 .... t ant~ pH<2DPAS V TLC/FID

Labile+polar org.Cu Nonpolar org.Cul I

Liplid Base Aci~ Residue -mobile -mobile

material material

Fig. 2. Protocol for determination of copper speciation, chemical, and ecological parameters in seawater.

362 r . c . H U N G A N D B.C. H A N

The classes of organic compounds such as lipids, acid-mobile and base- mobile compounds, collected from sea water by the C-18 Sep-Pak technique, were further separated by TLC with chloroform/chloroform/methanol/ ethylamine (10:15:1), and methanol/formic acid/water (5:5.1:1) solutions, respectively. Extra-pure phytol, n-octylamine and suberic acid obtained from Sigma chemical company were used as the standards of lipids, acid-mobile compounds and base-mobile compounds, respectively. To study the bioac- cumulation of copper by the oysters, cultures of approximately 5000-6000 individuals (age 1 month) were suspended at three stations in the Charting coastal area (Fig. 1) from December 1988 to March 1989. Fifteen to 20 in- dividual oysters of similar size were sampled on Day 0 to obtain initial dry weight and tissue copper concentration. During the next 3 months, 15-20 oysters at different time intervals were sampled from each station and analyz- ed for the copper content in the whole soft part. NBS oyster tissue standard (NBS 1566, USA) was used as control.

Oyster samples were digested with a mixture of nitric and sulfuric acid (1:1, v/v) solution, and the supernatant analyzed for copper by atomic ab- sorption spectrophotometry.

RESULTS AND DISCUSSION

Copper species and forms

Dissolved and particulate forms of copper in the sea water along the Char- ting mariculture area varied with location (Table 1). For instance, a relatively high concentration of dissolved copper (range from 4.03 to 28.6/~g/1) was generally found at the nearshore stations (stations 2-8) compared with those of the offshore stations (stations 9-12). For the particulate copper, the highest concentrations were observed at station 1, in the Erhjin River. The offshore stations also contained relatively lower concentrations of par- ticulate copper than the nearshore and estuarine stations. It was very in- teresting to note that about 100% of non-labile copper in the total dissolved copper was obtained at station 1 in October and station 9 in December. This high content of non-labile organic copper might be caused by the high con- centrations of both dissolved organic pollutants and dissolved copper discharged from local cities and industries. Table 1 showed that the contents of polar (range from 0.18 to 9.33/zg/1) and non-polar (range from 0.12 to 13.9 /zg/1) copper varied with location. In general, relatively low polar organic copper (less than approx. 48% of the copper organic complexes) was found in the Charting coastal water. This result was in accordance with that of Perrish et al. [6], indicating that non-polar organic compounds from man- made organic pollutants were the major organic complexes in the study area.

COPPER POLLUTION IN THE MARICULTURE AREA OF TAIWAN

TABLE 1

Concentrations of copper species (~g/1) in seawater along the Charting coastal area

363

Sampling Station Total Parti- Dissolved date culate

Total Labile Non-labile

Inor- Free a Total Polar Non- ganic polar

Oct. 11, 1988

Dec. 15, 1988

1 739 723 15.5 ND ND 15.5 ND ND 2 9.97 3.25 6.72 1.81 0.03 4.88 3.39 1.49 3 23.6 5.33 18.2 10.8 0.19 7.28 1.81 5.47 4 10.3 4.26 6.00 2.62 0.05 3.33 0.18 3.12 5 14.1 4.13 I0.0 2.35 0.04 7.62 1.38 6.24 6 17.9 6.90 11.0 5.20 0.09 5.73 3.71 2.02 7 16.4 5.61 10.8 5.21 0.09 5.45 3.25 2.20 8 21.2 4.84 16.4 8.42 0.15 7.79 6.25 1.54 9 8.11 3.09 5.02 1.44 0.02 3.56 3.45 0.12

10 12.1 4.25 7.82 4.99 0.09 2.74 1.09 1.65 11 11.5 3.11 8.40 4.75 0.08 3.57 0.44 3.13 12 10.6 4.04 6.51 3.17 0.05 3.29 0.58 2.71

1 186 165 20.7 ND ND 20.7 9.33 11.4 2 30.5 11.5 19.0 8.48 0.15 10.4 8.21 2.19 3 25.2 3.13 22.1 12.7 0.22 9.24 8.14 1.10 4 15.9 5.93 9.97 6.62 0.11 3.24 1.41 1.83 5 25.9 9.57 16.4 7.79 0.13 8.43 5.23 3.20 6 11.0 2.54 8.46 3.38 0.06 5.02 2.15 2.87 7 4.78 0.75 4.03 1.01 0.02 3.00 0.48 2.52 8 29.5 0.90 28.6 8.77 0.15 19.6 5.71 13.9 9 6.55 1.42 5.13 ND ND 5.13 3.24 1.89

10 11.0 1.07 9.93 3.23 0.06 6.64 4.12 2.52 11 18.1 5.29 12.8 5.29 0.09 7.45 4.07 3.38 12 9.38 2.45 6.93 3.17 0.05 3.71 1.54 2.17

aFree copper ion (Cu 2+) calculated by the method obtained by Hanson et al. by (see Ref. 13). ND, not detected.

Copper complexing capacity and organic compounds

I n t h e C h a r t i n g c o a s t a l a r e a , c o p p e r c o m p l e x i n g c a p a c i t y r a n g e d f r o m 1.38

to 7 . 07 /~g Cu/1 ( T a b l e 3). F i g u r e 3 s h o w e d a n e x a m p l e o f t h e t i t r a t i o n o f a

f i l t e r e d w a t e r s a m p l e c o l l e c t e d f r o m s t a t i o n 4 in t h e C h a r t i n g c o a s t a l a r e a .

A c o p p e r c o m p l e x i n g c a p a c i t y v a l u e o f 7 . 07 /~g C u / l w a s o b t a i n e d f r o m t h e

364 x.c..UNG ANO a.C. ,AN

TABLE 2

Concentrations of copper (#g/l) in seawater along the mariculture areas

Sampling Station Total Parti- Dissolved date culate

Total Labile Non-labile

Inor- Free a Total Polar Non- ganic polar

Oct. 11, 1988 S1 14.1 4.13 10.0 2.35 0.04 7.62 1.38 6.24 Dec. 25, 1988 25.9 9.57 16.4 7.79 0.13 8.43 5.23 3.20 Jan. 11, 1989 27.1 3.52 23.6 2.04 0.03 21.5 7.82 13.7 Jan. 25, 1989 23.9 2.11 21.8 13.1 0.23 8.48 0.80 7.68 Feb. 16, 1989 15.3 3.98 11.3 2.32 0.04 8.91 2.39 6.52 Mar. 15, 1989 10.6 5.51 5.12 1.70 0.03 3.39 2.83 0.56

Oct. 11, 1988 $2 23.6 5.33 18.2 10.8 0.19 7.28 1.81 5.47 Dec. 25, 1988 25.2 3.13 22.1 12.7 0.22 9.24 8.14 1.10 Jan. 11, 1989 9.11 1.25 7.86 3.24 0.06 4.62 2.03 2.59 Jan. 25, 1989 10.2 5.16 4.99 2.12 0.04 2.87 1.09 1.78 Feb. 16, 1989 10.2 2.49 7.71 2.15 0.04 5.56 3.81 1.75 Mar. 15, 1989 9.22 3.19 6.03 2.50 0.04 3.83 2.64 1.19

Dec. 25, 1988 $3 10.2 2.08 8.07 ND ND ND ND ND Jan. 11, 1989 11.4 1.09 10.3 2.76 0.05 7.55 2.81 4.74 Jan. 25, 1989 9.3 3.10 6.24 2.87 0.05 3.37 1.59 1.78 Feb. 16, 1989 6.5 1.54 4.97 3.03 0.05 1.94 1.59 0.35 Mar. 15, 1989 9.68 3.06 6.62 2.42 0.04 4.20 0.73 3.47

aFree copper ion (Cu 2÷) calculated by the method obtained by Hanson et al. (see Ref. 13). ND, not detected.

intercept on the abscissa through a least squares regression line of Y (peak current) versus X (copper spike). The correlation coefficients of copper com- plexing capacity and DOC in the nearshore and offshore stations were respectively 0.78 and -0.76. the high positive correlation coefficients in the nearshore water might be due to the same characteristic of organic com- pounds [7].

Figure 4 showed that lipids (range from < 6.14/~g/1 to 5.07 mg/l), base- mobile compounds (range from 0.03 to 9.33 mg/1), and acid-mobile com- pounds (range from < 9.35/~g/1 to 8.01 mg/1) varied with location. For in- stance, extremely high levels of acid- and base-mobile organic compounds, were generally observed at the river station (station 1), and the next highest

COPPER POLLUTION IN THE MARICULTURE AREA OF TAIWAN 365

TABLE 3

Concentrations of chemical, ecological parameters and copper complexing capacity (CC) in waters along the Charting coastal area

Date Station POC DOC Nitrate Phos- Chl. a ATP CC (mg/l) (mg/1) (~M) phate (#g/l) (ng/1) (tzg Cu/1)

(~M)

July, 1988

Oct., 1988

Dec., 1988

1 11.8 16.7 2.85 18.6 8.31 5.59 ND 2 0.62 0.85 1.04 0.71 5.77 0.87 ND 3 0.36 0.69 1.20 0.35 3.76 1.11 ND 4 0.31 0.56 1.09 0.40 3.06 0.65 ND 5 0.24 1.31 0.82 0.30 4.37 0.68 ND 6 0.80 1.90 1.23 0.66 11.7 1.39 ND 7 0.80 2.50 0.93 0.30 6.21 0.70 ND 8 0.47 1.83 1.13 0.40 5.25 1.38 ND 9 0.49 1.33 0.96 0.30 2.54 0.70 ND

10 0.41 0.57 1.00 0.35 4.72 1.02 ND 11 0.37 0.59 1.16 0.30 4.55 0.67 ND 12 0.40 0.36 1.10 0.30 5.25 0.68 ND

1 5.22 4.58 ND 8.73 4.00 0.43 ND 2 0.66 0.33 1.32 0.77 2.31 0.06 5.58 3 0.78 0.53 2.83 0.24 1.47 0.17 ND 4 0.37 0.18 2.45 1.25 1.53 0.06 7.07 5 0.49 0.22 7.71 1.03 1.78 0.06 2.63 6 0.48 0.38 3.68 1.92 1.72 0.07 2.85 7 0.42 0.34 2.39 0.92 2.11 0.05 3.12 8 0.22 0.28 1.26 0.71 2.31 0.09 1.38 9 0.24 0.24 1.04 0.82 1.83 0.05 3.39

10 0.36 0.25 2.91 0.92 1.86 0.04 2.19 11 0.21 0.40 2.85 0.28 2.00 0.05 5.82 12 0.37 0.18 2.52 0.45 1.89 0.13 3.52

1 2.20 12.8 4.59 21.2 0.33 4.61 ND 2 0.80 4.10 3.27 6.84 1.39 4.72 ND 3 0.24 0.83 3.08 1.61 0.94 2.40 ND 4 0.21 1.60 2.04 0.87 1.28 2.33 ND 5 0.21 1.38 2.12 0.51 0.97 3.05 ND 6 0.11 0.60 1.40 0.19 0.92 1.27 ND 7 0.27 0.79 1.86 0.24 0.75 1.16 ND 8 0.14 0.58 2.02 0.35 0.69 1.02 ND 9 0.12 0.64 2.19 0.56 0.50 0.86 ND

10 0.36 1.07 2.77 0.77 0.50 1.31 ND 11 0.38 0.73 1.77 0.92 0.94 1.11 ND 12 0.46 0.67 1.93 0.45 0.36 1.37 ND

ND, not determined.

366

,.-1

<

I!

oo

° ~

. .=

0)

o

" o

,..a

<

©

©

z ~

T.C. H U N G A N D B.C. HAN

~ d d d ~

I I

I I

I I I I

£ d d d d d d d d ~ I I I l l

I I I I I

I I I I

I I I I I I

I I I I I I

I I I I I I I

=

m ~

V V

COPPER POLLUTION IN THE MARICULTURE AREA OF TAIWAN 367

<

i

~L

0

~)

=

~.)

0

..-.

~ 8

<

<

~J

0

L) ©

I

i i

i i i

i i i

i i i i i i i I

i i i i i i i i I

. . d

V V

a ,

368 T.C. HUNG AND B.C, HAN

100

8O

~ 6 0

~o

~. 20

Y = 2 . 7 2 x - 1 9 , 2 4

R = 0 . 9 9 9

C C. = 7. 07,ug Cu/I

~ . ..~ ~ ~ o ~ O~"

0 ~ '

0 I0

o ~ f

1"

% I "

f / o f

.o f

.o"

I I I i i I

20 30 40

Copper added (ppb)

Fig. 3. Copper complexing capacity ot sea water (CC) collected from the Charting coastal area at station 3, October 1988.

concentrations were found in the Erhjin Chi estuary (station 2). These high contents o f organic compounds obtained at the nearshore stations might be due to the organic pollutants discharged from land sources. However, the lipid distribution gave the opposite phenomenon. Lower lipid concentrations were generally observed at the nearshore stations, particularly at the river

DOM(ppm) ~~....{.-. ~ ~{(.,'-...

, o.o ~ ....... • .~:.:%..:;.:~:.::..:;.::::.::.::::~:.:::~:.::::.;:~::~:.::;:.::.:.:.::~;~:~:.~.::.::~.~.;:::..:::;.~.::~..~::.:~;~.::~:~ .~....;.,~~~.

4.0 ii'.i.i.i'i '.i:i'.i':'-i'ii('i'.'{:ii .ii'.i.ii.i'i " .i-"i,i."i:'.".,".".".i"""i"~:'.

~..~i.:.:.:..:.:,:,.:...~.,:~:.:,:..~..:.:.~~~..~.-~ ~ :::::::::::::::::::::::::::::::::::::::: .o

~r~ ',~ ",."::.:. ''<{%;.>;.;,.. %ISS;-..;.:. :<..:~.:.:-:,.

~ ~ . W ' t . ~ ....... x , ~ % "%" "~',~

% %

Fig. 4. Seasonal variations of dissolved organic matter in water samples collected from the Charting coastal area in 1988.

COPPER POLLUTION IN THE MARICULTURE AREA OF TAIWAN 369

station, compared with those concentrations at the offshore stations. This phenomenon might indicate that lipids originating from dead organisms have already been decomposed and dissolved in the marine waters. High con- centrations of lipids not only gave a signal of organic pollution but also an indicator of high biological activities. Kennicutt and Jeffrey [8] reported that the high concentration of lipids (as high as 1 mg/l) in the polluted water of Tokyo Bay was observed. However, in the Charting coastal mariculture water, extremely high concentrations of lipids (> 2 mg/1) were generally found in July.

Copper bioaccumulation and elimination in oysters

The field experiments for determining the rate of copper accumulation in oysters were carried out and the results (Table 2, Fig. 5) indicated that the rate of copper accumulation varied with the original concentration of copper in oysters. For example, the initial rate of 770 #g/g per day was obtained at the Erhjin River estuary (station S1) when the copper content in oysters was 100/xg/g (dry weight). However, when the copper content was increased to 3528/zg/g, the copper bioaccumulation rate was decreased to 62.4/zg/g per day. Furthermore, the initial bioaccumulation rate was also varied with the

1 0 4 ~ I,~ I~ ~-~J I x I " f ' , ,

I- I I ' ~"~ I I - - -

I / " - -

b io a

E Ltll o s 7

tl/ , s 2 I , ,'53, , I

1020 30 50 90 120 150

Exposure time (deys)

Fig. 5. Bioaccumulation of copper (ppm, dry weight) in oysters (Crassostrea gigas) cultured along the Charting coastal area (S 1: Erhjin Chi estuary, $2: Charting coastal station, and $3: Outlet of Shindai Power Plant). The initial bioaccumulation rate, V~ = 770 ppm/day, V2 = 590 pprn/day, and V 3 = 330 pprn/day for S1, $2, and $3, respectively.

370 T.C. HUNG AND B.C. HAN

1800

1500

"•1200 ~ 9oo 0

~d 5OO

0 £3 300

I0

vo

0 i I I Z lO 0 10 20 30

Exposure t ime (days)

Fig. 6. Elimination of copper (ppm, dry weight) in oysters (Crassostrea gigas) cultured at the Erhjin Chi estuary in February 1987. The initial elimination rate, V 0 = 300 ppm/day, and final elimination rate, V t = 5 ppm/day.

ambient concentrations of copper species, mainly particulate copper. Figure 5 also indicated that the initial bioaccumulation rates of 770, 590 and 330 /~g/g per day were obtained in the mariculture water containing the par- ticulate copper of 9.57, 3.13 and 2.08/~g/1, respectively at the Erhjin River estuary (S1), Charting coastal area ($2) and the Outlet of Shindai Power Plant ($3). The higher the concentration of particulate copper, the higher the copper bioaccumulation rate in oysters that was observed. The experimental results strongly suggest that when the copper content was over 500/~g/g (dry weight), the color of oysters became green.

The elimination of copper in oysters was also studied. Figure 6 showed that green oysters having high copper content of 2225/~g/g (dry weight) were cultured in the laboratory at room temperature. The culture medium, con- taining a total copper concentration of less than 1/xg/1, was collected from off the eastern coast of Taiwan. High initial rate of 300 ppm Cu/day was observed with a 4-day incubation. After a 14-day incubation, the color of oysters returned to normal and the final elimination rate dropped to 5/xg/g per day.

Relationships among these parameters

The uptake of copper in oysters has been found to be influenced by physical, chemical and ecological parameters [5,9]. Table 3 showed the

COPPER POLLUTION 1N THE MARICULTURE AREA OF TAIWAN 371

chemical and ecological parameters and analytical data of the sea water along the Charting coastal area including the Erhjin River. The distribution of chlorophyll a (range from 0.33 to 11.70/~g/l), ATP (from 0.04 to 5.59 ng/1), DOC (from 0.18 to 16.7 mg/1), POC (from 0.12 to 11.8 mg/l) and nutrients (such as nitrate and phosphate) varied with location. In general, higher values of these chemical and ecological parameters were obtained at the nearshore stations, particularly at the river station, than at the offshore sta- tions [5].

In order to investigate the influence of environmental factors on oyster bioaccumulation of copper, the correlations among the chemical and ecological parameters including the classes of organic compounds and the species of copper in the Charting coastal area were calculated. At the near- shore stations, there are significant correlations (over 0.77) among lipids ver- sus labile, non-labile, non-polar and bioavailable (sum of particulate and labile) copper and DOC; base-mobile organic compounds versus DOC. At the offshore stations, the correlation coefficients of lipids versus POC, labile, non-labile, non-polar and bioavailable copper; base-mobile compounds ver- sus DOC decreased to 0.70, -0.36, -0.45, 0.71, -0.35, and 0.83, respectively.

SUMMARY

At station l, copper was almost entirely in the particulate phase whereas at other stations, dissolved organic copper predominated as a rule. At station l, dissolved copper was entirely non-labile on both dates, while at the re- maining locations, the labile dissolved fraction was usually important also. High correlation coefficients were observed between base-mobile compounds and chlorophyll a (0.85), base-mobile compounds and DOC (0.68). The results may explain why the water column is dominated by organic matter produced autochthonously by the growth and decomposition of phytoplankton.

At stations S1, $2 and $3, the bioaccumulation data indicate that the total uptake of copper per oyster is an exponential function of exposure time for the first 2 weeks with an accumulation rate of 214, 150 and 73.8/~g/g per day, respectively, and then levels off. These results could be explained by the copper accumulation capacity of oyster approaching saturation. It is worth noting that depuration of copper by green oysters appeared to decrease ex- ponentially with time for the first 1-23 days.

REFERENCES

1 T.C. Hung, P.G. Meng and C.C.H. Tsai, Distribution of copper in the Taiwan mariculture area. J. Environ. Soc. ROC, 10 (1987) 13.

372 T.C. HUNG AND B.C. HAN

2 T.C. Hung, Heavy metal pollution and marine ecosystem as a case study in Taiwan, in R. Abbou (Ed.), Hazardous Waste: Detection, Control and Treatment, Elsevier, New York, 1988, p. 869.

3 C.R. Chen, B.C. Han and T.C. Hung, Copper complexing capacity in the coastal water of Taiwan. Bull. Inst. Chem., Acad. Sin., 35 (1988) 71.

4 K.S. Chen, B.C. Han and T.C. Hung, Organic copper in the coastal water of Taiwan. Bull. Inst. Chem., Acad. Sin., 36 (1989) 105.

5 T.C. Hung, A. Chuang, B.R. Chou and T.L.Cheng, Relationships among particulate organic carbon, chlorophyll a, and primary productivity in the sea water along Taiwan western coast. Acta Oceanogr. Taiwanica, 13 (1982) 109.

6 C.C. Perrish, R.P. Delmas, P.J. Wangersky and R.G. Ackman, Iatrosan measured pro- files of dissolved and particulate marine lipid classes over in Scotian Slope and in Bedford Basin. Mar. Chem., 23 (1988) 1.

7 A.D. Newell and J.G. Sander, Relative copper binding capacities of dissolved organic compounds in a coastal plain estuary. Environ. Sci. Technol., 20 (1986) 817.

8 M.C. Kennicutt and L.M. Jeffrey, Chemical and GC/MS characterization of marine dissolved lipids. Mar. Chem., 10 (1981) 367.

9 T.C. Hung, C.Y. Kuo and M.H. Chen, Mussel watch in Taiwan, ROC (2): Seasonal bioaccumulation factors of heavy metals. Bull. Inst. Chem., Acad. Sin., 30 (1983) 49.

10 T.C. Hung, B.C. Han and S.J. Wu, Green oyster: species and forms of copper in the Charting coastal water. Acta Oceanogr. Taiwanica, 23 (1989) 33.

11 T.C. Hung, B.C. Han and C.Y. Horng, A case study of green oysters: dissolved organic matter in the Taiwan coastal water. Acta Oceanogr. Taiwanica, 24 (1989) 76.

12 B.C. Han and T.C. Hung, Green oysters caused by copper on the Taiwan coast. Environ. Pollut., 65 (1990) 347.

13 A.K. Hanson, C.M. Sakamoto-Arnold, D.L. Huizenga and D.R. Kester, Copper com- plexation in Sargasso Sea and Gulf Stream Warmcore ring water. Mar. Chem., 23 (1988) 181.