Biomonitoring of heavy metals and trace organics using the intertidal mussel Perna viridis in Hong Kong coastal waters

This paper presents the results of a 6-year (1998–2003) survey of trace toxics in the intertidal mussel Perna viridis conducted by the Hong Kong Environmental Protection Department. Concentrations of heavy metals and trace organics were measured in the soft bodies of P. viridis collected from five sites in Hong Kong waters, i.e. Wu Kai Sha (Tolo Harbour), Ma Wan (Northwest), Tsim Sha Tsui (Victoria Harbour), Tai Tam (Hong Kong South) and Lamma Island (Southern Waters) in order to establish the spatial patterns of contaminants in mussels. Among the metals analysed, Cd showed a significant concentration gradient in Hong Kong waters. The levels of Cd in P. viridis were significantly higher at Ma Wan as compared to the other sites studied. Ma Wan also had relatively higher concentrations of Pb. Mn concentrations were particularly prominent at Wu Kai Sha. Significantly higher concentrations of Hg and Cu were recorded at Tai Tam and Tsim Sha Tsui. Tai Tam and Wu Kai Sha had higher levels of V; whereas higher Ni concentrations were recorded at Lamma Island and Tai Tam. No clear spatial patterns for Al, As, Cr, Fe and Zn were observed.

Higher concentrations of PAHs in P. viridis were observed around urban centres impacted by sewage discharges (e.g. Tsim Sha Tsui); whereas higher PCB levels were found not only in Tsim Sha Tsui but also in less urbanised areas such as Lamma Island and Tai Tam, suggesting that these may be due to non-sewage related inputs. The study also shows that Northwest and Southern waters are subject to a higher degree of DDT pollution compared with other sites. Of the 17 dioxin compounds analysed, positive data were mostly recorded for two compounds which are of low toxicity (i.e. OCDD and 1,2,3,4,6,7,8-HpCDD) whereas the most toxic congeners (i.e. 2,3,7,8-TCDD and 1,2,3,7,8-PeCDD) were not detected in the 6 years of monitoring. In general, the levels of OCDD in P. viridis were found to be higher in Tai Tam and Lamma Island in Southern Waters of Hong Kong.

This study found that the levels of some highly toxic heavy metals (i.e. Cd, Hg and Pb) in the mussel P. viridis did not exceed the recommended limits for shellfish as food in Hong Kong (i.e. Cd: 2.0 ppm; Hg: 0.5 ppm; Pb: 6.0 ppm wet weight). The levels of As and Ni in P. viridis were also well below the action limits set by the US FDA (i.e. As: 86 ppm; Ni: 80 ppm wet weight). DDT and PCB contaminations in P. viridis were below the concentrations of concern. Compared with data obtained in the 1980s, the current levels of DDTs in P. viridis were 4–16 times lower; whereas Pb concentrations recorded in Tsim Sha Tsui have also been lowered significantly. This is mainly related to reduction in local and regional pollution sources in the past 20 years.     

Pilot Experiments on Planting Seedlings and Small Seagrass Propagules in Western Australia

The aim of this study was to investigate characteristics of some south-western Australian seagrasses that might be useful for further investigation with respect to restoration. Although seeds and fruits of seagrass species may be useful for producing propagules for planting, their collection is based on knowing when they become available. Posidonia australis, P. angustifolia and P. coriacea flower profusely annually, but P. sinuosa does not. Once collected, the fruits dehisce in a few days and are ready for planting. It was found that the best way to plant Posidonia seedlings is in clusters in Growool^® blocks but the rhizomes do not spread quickly. Single Posidonia seedlings, when planted out, were not successful and neither were naturally colonizing seedlings of Posidonia sinuosa in unexposed unvegetated areas. Successful planting of seedlings and sprigs of Amphibolis in matting needs months of calm weather for them to spread out from their attaching matting which must be very securely attached to the substratum. Little success was achieved in growing Amphibolis. In a search for Halophila ovalis seeds, few were found because of the patchy nature of their fruiting and small size. Germination was slow but 63% germinated within 6 months.     

The potential role of mucus in the depuration of copper from the mussels Perna viridis (L.) and Septifer virgatus (Wiegmann)

Experiments were conducted to measure the mucus secretion rate in two common mussels in Hong Kong, Perna viridis (L.) and Septifer virgatus (Wiegmann), when chronically exposed to Cu (50 μg l⁻¹). After a 3-month exposure period, the mucus production rate of P. viridis at 25°C in the metal treatment was 2.65 times that of the control (10.7 vs 4.0 mg g⁻¹ dry wt h⁻¹), while S. virgatus showed a 1.85 times difference (4.41 vs 2.38 mg g⁻¹ dry wt h⁻¹). Mucus secretion by P. viridis under acute Cu exposure (0.5 mg l⁻¹) was significantly higher in the metal treatment than the control (13.43 vs 9.16 mg g⁻¹ dry flesh wt). Metal contents of the mucus secreted was about 18 times those in the control and 6 times in the soft tissues. Therefore, mucus appears to be an effective agent for Cu depuration in the mussel. The significance of these results to the local distribution and abundance of the mussels is discussed.     

Hydrocarbon contamination on the Antarctic peninsula: III. The Bahia Paraiso—Two years after the spill

Two years after the release of 600 000 l of diesel fuel arctic into Arthur Harbor, little spill-related contamination can be detected in intertidal limpets (Nacella concinna) and subtidal sediments. Periodic releases of small amounts of material from the ship oil nearby islands, in particular the intertidal areas of Christine, Limitrophe and Humble Islands. Subtidal sediment contamination is primarily due to other local inputs such as ship, boating and station activities. Beaches were unusually contaminated after 2 yr, but quiescent weather conditions, occasional releases from the wreck, and prevailing currents may concentrate hydrocarbon contamination in relatively low energy areas. Intertidal limpets (N. concinna) collected along these beaches were also contaminated. The volatility of the fluid, the amount spilled, and the dynamic weather and current conditions in Arthur Harbor tended to minimize long-term contamination of the area.     

Heavy metal accumulation in the vicinity of a desalination plant

The brine effluent produced by a flash distillation desalination plant contains elevated levels of some heavy metals. The effluent produced from such a plant operating in Jersey during the summer months leads to increased metal concentrations in limpets and some Fucus species. This is most evident with copper which remains at elevated levels during the winter period of plant inactivity, probably due to the copper coating retained on boulders and pebbles on the upper shore.     

Bioavailability of dissolved organic phosphorus compounds to typical harmful dinoflagellate Prorocentrum donghaiense Lu

The bioavailability of dissolved organic phosphorus (DOP) compounds to harmful alga Prorocentrum donghaiense Lu and its alkaline phosphatase activity (APA) were studied. Results showed that P. donghaiense could utilize the test DOP compounds such as glucose-6-phosphate (G-6-P), adenosine triphosphate (ATP) and ribonucleic acid (RNA) to sustain its growth. Nutrition between the test DOP compounds and orthophosphate was comparable. P. donghaiense could utilize intracellular phosphorus to sustain growth under depletion of dissolved phosphorus. Variation of APA in different test DOP compounds was addressed; the controlling mechanism of APA in different DOP media was discussed.     

Responses of the green-lipped mussel Perna viridis (L.) to suspended solids

Laboratory experiments were conducted to investigate the lethal and sublethal effects of suspended solids on the survival and physiological, behavourial and morphological changes of the green-lipped mussel Perna viridis collected from Tolo Harbour, Hong Kong. Results showed that P. viridis survived in all test conditions of suspended solids from 0 to 1200 mg/l over a period of 96 h. Physiological responses of the green-lipped mussel under 14-d exposure of suspended solids from 0 to 600 mg/l, followed by 14-d recovery in natural seawater, revealed no significant changes (p>0.05) in oxygen consumption and dry gonosomatic index for treatments in different concentrations of suspended solids and exposure time. Changes in clearance rate were only found to be significant (p<0.001) with exposure time. Responses in behavourial and morphological changes of the green-lipped mussel were also studied under similar experimental treatments and exposure time. Byssus production was significantly (p<0.001) related to exposure time. Gill damage, however, was significantly greater in treatments (p<0.001). Present findings suggested that P. viridis could tolerate a high level of suspended solids in the laboratory. There were dose-dependent effects of suspended solids on morphology of gill filaments. Implications of survival and responses of the green-lipped mussel to suspended solids in the marine environment are discussed.     

Assessment of a Mediterranean shallow lentic ecosystem (Lake Pamvotis, Greece) using benthic community diversity: Response to environmental parameters

Macroinvertebrates play a key role in freshwater lentic and lotic ecosystems. The macroinvertebrate benthic community of a shallow Mediterranean lake (Lake Pamvotis, NW Greece) was studied. The benthic assemblage was sampled monthly at five sites during a period of 1 year (Apr. 1998–Mar. 1999). In addition hypolimnetic water quality variables were monitored over the same period at each site.

The aim of the study was (a) to describe the intra-annual and spatial variability in benthic communities, (b) to relate possible community changes to environmental conditions and (c) to evaluate the responses of the lake's ecological status on community indices.

The benthic fauna of Lake Pamvotis was found to be very limited with a total of 10 species belonging to five taxonomic groups. The oligochaete community comprised 80% of the total benthic fauna with Potamothrix bavaricus as a new record for the Lake Pamvotis and Potamothrix hammoniensis, being the dominant benthic species represented more than 61% of the total benthic fauna. Chironomus plumosus was the most abundant chironomid species contributing with about 6% of the total benthic fauna, and Chaoborus flavicans with 19% was the important dipteran. Almost all benthic species showed the same intra-annual seasonal pattern, with peak population densities during spring and early summer except P. hammoniensis which predominated during the whole sampling period. Dissolved oxygen and temperature seemed to be the main environmental factors affecting community indices.

Benthic communities are affected by human disturbances in Lake Pamvotis shifting their composition to more tolerant taxa, reflecting also the eutrophic to hypertophic character of the lake.     

Trace Metals in Selected Benthic Invertebrates from Belgian Coastal Waters (1981–1996)

Mercury, cadmium, lead, copper, zinc and chromium were assessed in brown shrimp (Crangon crangon), swimming crab (Liocarcinus holsatus), hermit crab (Pagurus bernhardus), starfish (Asterias rubens) and cut trough shell (Spisula subtruncata) from ten sampling stations off the Belgian coast, including three dredge spoils disposal sites. Calculations of time trends indicated that on the whole concentrations of trace metals had significantly decreased since 1981, also on dredged material dumping sites. Starfish and hermit crab were found to be the most promising benthic indicator species for metal contamination. Globally, the location of the sites, including the dredge spoils disposal zones, showed to be of little influence on the concentrations of trace metals.     

The Effects of Mercury Exposure on the Surface Morphology of Gill Filaments in Perna perna (Mollusca: Bivalvia)

This study investigated the possibility that changes in the surface morphology of mussel (Perna perna) gill filaments may be used to indicate the relative toxicity of pollutants in the marine environment. Healthy, adult P. perna were collected and immersed in 2 free-flow tanks. Mercury was added to seawater as it entered tank 1 to achieve a constant level of 50 μg/l⁻¹ over 24 days. Uncontaminated seawater was circulated over the mussels in tank 2 (control) for the same period. A 25 mm² area of gill filament was removed from each of the 5 specimens before and after 24 days immersion in tank 2, and after immersion for 1, 2, 4, 8, 16 and 24 days in tank 1. These were examined using a scanning electron microscope. The remaining soft tissues from each animal were analysed for mercury using an atomic absorption spectrometer. Mercury concentration increased from 0.13 (pre-immersion) to 87 μg/g after day 24. Surface morphology remained normal for all animals in tank 2 and for those exposed to Hg for up to 8 days. However, from 16 to 24 days exposure there was a gradual increase in the diameters of microvilli, a depletion of abfrontal cilia, an increase in abnormal, perhaps necrotic cells and an unusual increase in the number of cilia on the lateral surfaces. These results confirm that P. perna is an efficient bio-accumulator and suggest that their gill pathomorphology may be a useful indicator of toxicity.     

Cardiac and Lysosomal Responses to Periodic Copper in the Mussel Perna viridis (Bivalvia: Mytilidae)

Biological monitoring of metals in marine waters often employs tissue residue analyses from bivalves. Chemical residues in tissues do not elucidate biological injury, however, whereas biomarkers, do afford sensitive stress indices. Physiological and cytological responses were evaluated in laboratory experiments from the mussel Perna viridis in order to assess the species’ suitability for pollution monitoring in the tropics. Periodic copper exposure increased tissue residues but failed to perturb cardiac activity. Copper destabilized haemocyte lysosomal membranes in a exposure-dependent relationship showing a sensitive, rapid, indication of stress. Cytological biomarkers from P. viridis have potential as indices of pollutant exposure and could facilitate effective biomonitoring of water quality throughout South-east Asia.     

Towards a more ecologically relevant assessment of the impact of heavy metals on the photosynthesis of the seagrass,Zostera capricorni

This in situ study used photosynthetic activity (measured as chlorophyll a fluorescence) and photosynthetic pigment concentrations to assess the effect of copper, cadmium, lead and zinc on the seagrass Zostera capricorni. Custom-made portable in situ exposure (PIE) chambers were developed so seagrasses could be dosed within the meadow. Z. capricorni was exposed to 0.1 and 1 mg l⁻¹ of metal solutions for 10 h. During this time and for the subsequent four-day recovery period, the effective quantum yield of photosystem II (PS II) (ΔF/Fm^′) was measured. While the results were variable, copper and zinc exposed samples had a depressed ΔF/Fm^′ during the exposure period. Samples exposed to zinc recovered to pre-exposure levels but those exposed to copper did not. Cadmium and lead did not impact on the chlorophyll a fluorescence and the chlorophyll pigment data supported these findings. This study presents an innovative new application of chlorophyll a fluorescence stress assessment.     

Effect of lethal concentrations of copper on Mystus bleekeri (day)

In the batch test with daily exchange of medium the effect of copper ions at 2.4 mmol/l (Ca²⁺ + Mg²⁺) over 10 d of exposure is investigated. The LT₅₀ values and 95% confidence limits at 3.7, 2.1 and 1.0 ppm of Cu were 26.7 (23.6… 29.9), 47.5 (39.6… 57.0) and 101.5 (78.5… 130.7) h, respectively. The LC₅₀ values of copper in ppm were 4.05 (24 h), 1.69 (48 h), 0.829 (9.96 and 0.776 (240 h).     

Effect of an acid mine drainage effluent on phytoplankton biomass and primary production at Britannia Beach, Howe Sound, British Columbia

We investigated the effect of acid mine drainage (AMD) from an abandoned copper mine at Britannia Beach (Howe Sound, BC, Canada) on primary productivity and chlorophyll a levels in the receiving waters of Howe Sound before, during, and after freshet from the Squamish River. Elevated concentrations of copper (integrated average through the water column >0.050 mg l⁻¹) in nearshore waters indicated that under some conditions a small gyre near the mouth of Britannia Creek may have retained the AMD from Britannia Creek and from a 30-m deep water outfall close to shore. Regression and correlation analyses indicated that copper negatively affected primary productivity during April (pre-freshet) and November (post-freshet). Negative effects of copper on primary productivity were not supported statistically for July (freshet), possibly because of additional effects such as turbidity from the Squamish River. Depth-integrated average and surface chlorophyll a were correlated to copper concentrations in April. During this short study we demonstrated that copper concentrations from the AMD discharge can negatively affect both primary productivity and the standing stock of primary producers in Howe Sound.     

Physiological Responses of Five Seagrass Species to Trace Metals

Trace metal run-off associated with urban and industrial development poses potential threats to seagrasses in adjacent coastal ecosystems. Seagrass from the largest urban (Moreton Bay) and industrial (Port Curtis) coastal regions in Queensland, Australia were assessed for metal concentrations of iron (Fe), aluminium (Al), zinc (Zn), chromium (Cr) and copper (Cu). Trace metal concentrations in seagrass (Zostera capricorni) leaf and root-rhizome tissue had the following overall trend: [Fe] > [Al] > [Zn] > [Cr] > [Cu]. Rainfall events and anthropogenic disturbances appeared to influence metal concentrations in seagrasses with the exception of Al, which does not appear to bioaccumulate. In laboratory experiments, five seagrass species (Halophila ovalis, H. spinulosa, Halodule uninervis, Z. capricorni, Cymodocea serrulata) were incubated with iron (1 mg Fe l⁻¹) and copper (1 mg Cu l⁻¹) and responses assessed by changes in PSII photochemical efficiency (Fv/Fm), free amino acid content and leaf/root-rhizome metal accumulation. Iron addition experiments only affected Halophila spp, while copper additions affected other seagrass species as well. Trace metal contamination of seagrasses could have ramifications for associated trophic assemblages through metal transfer and seagrass loss. The use of photosystem II photochemical efficiency as well as amino acid concentrations and composition proved to be useful sublethal indicators of trace metal toxicity in seagrasses.     

Cultured red alga to measure pollution

The red alga Callithamnion grew equally well in seawater samples from unpolluted and polluted localities. The species was insensitive to nitrate and phosphate levels of the medium and thus cannot be used as an indicator of eutrophication. Relatively small copper concentrations depressed the growth of the species. No difference in copper tolerance was demonstrated between isolates from unpolluted north-east Scotland and polluted County Durham, but the isolate from unpolluted south-west England was apparently less resistant to copper.     

Effect of Copper on Activity and Feeding in the Subtidal Prosobranch Babylonia lutosa (Lamarck) (Gastropoda: Buccinidae)

Individuals of the subtidal gastropod Babylonia lutosa were exposed to three sublethal concentrations of copper (0.02, 0.06 and 0.20 ppm) for four weeks. The percentage of normally active individuals (body extended and moving with the foot attached) decreased and the percentage of snails retracted (body extended, foot not attached, unable to move) and in a distressed stage (body retracted within the shell) increased with increase in copper concentration. Exposure to copper also decreased significantly the ability of an individual to move, and to burrow, but the effect was only observed in week 1, individuals were fully recovered in week 2. Feeding, however, was not affected, both in terms of the percentage number of individuals which fed, and the time spent feeding. Results were discussed with respect to the potential of using this animal as a bioassay tool in monitoring the effect of environmental perturbations on the subtidal community in Hong Kong.     

Nitrogenous Excretion of Penaeus monodon Postlarvae Fed with Different Diets

This study is designed to determine ammonia and nitrite excretion by Penaeus monodon 25–30 day old postlarvae fed with different diets (artificial and natural). Shrimps were fed with Chaetoceros calcitrans (phytoplankton), chironomid larvae (benthos), Brachionus sp. (rotifer) and artificial diet. In the control treatment, shrimps were not fed. The highest concentration of ammonia was excreted by shrimps fed with artificial diet and the lowest by that of control treatment. All treatments except where shrimps fed on C. calcitrans showed a sudden increase in ammonia-N excretion after 36 h. C. calcitrans contributed to high ammonia excretion only after 61 h. Shrimps fed with Brachionus sp. (rotifer) showed significantly higher nitrite excretion rate compared to other treatments. Shrimps fed on C. calcitrans depicted the lowest nitrite excretion. This experiment indicated that nitrogenous excretion was predominant in shrimps fed with artificial diets. Live food like algae and chironomids, despite a high protein content, contribute to low nitrogenous excretion and hence pose less adverse effects on water quality compared to artificial diets.     

Calcium alginate immobilized marine microalgae: experiments on growth and short-term heavy metal accumulation

Growth of 11 calcium alginate immobilized marine microalgal species belonging to eight taxonomical groups has been checked in the present work. Cellular densities inside the calcium alginate beads were monitored during 17 days. Good growth and maintenance of the structure of the beads were both found for some of the assayed species. One of those species (Tetraselmis chui, Prasinophyceae) was selected in order to perform a short term (up to 24 h) heavy metal accumulation experiment. Beads of calcium alginate containing (or not) cells of T. chui were exposed to 820 μg L⁻¹ Cu and 870 μg L⁻¹ Cd separately during a 24 h period, and accumulation of heavy metals in the beads was measured after this time and compared. Concentration of each metal in the supernatants was monitored at 5, 10, 60 min and 24 h from the beginning of the experiment. After 24 h, practically all Cu was removed by the beads. Beads with immobilized algae removed around 20% of total Cd, while beads without algae removed half of that percentage.