Barnacles as biomonitors of trace metal availabilities in Hong Kong coastal waters: changes in space and time

The use of selected organisms as biomonitors of trace metal bioavailabilities allows comparisons to be made over space and time. The concentrations of 11 trace metals (arsenic, cadmium, chromium, cobalt, copper, iron, lead, manganese, nickel, silver, zinc) were measured in the bodies of two barnacle species, Balanus amphitrite and Tetraclita squamosa, from up to 18 littoral sites from Hong Kong coastal waters in April 1998. These data provide evidence on the geographical variation in metal bioavailabilities at this time, and are compared selectively against historical data sets for 1986 and 1989. Geographical variation in bioavailabilities is clear for several metals, with hotspots for arsenic, copper, nickel and silver at Chai Wan Kok, and for lead in Junk Bay. Victoria Harbour sites head the rankings for silver and arsenic, and Tolo Harbour sites exhibit relatively elevated cobalt, manganese and zinc. Many bioavailabilities of trace metals to barnacles are lower in Hong Kong coastal waters in 1998 than in 1986. The two barnacle species are widespread and the extensive data set presented is a benchmark which can be compared to the results of similar biomonitoring programmes elsewhere in the Indo-Pacific and beyond.     

海洋沉积物中碱性磷酸酶活力与水体污染的关系

厦门西港和香港维多利亚港近岸沉积物中碱笥磷酸酶活力与水体受污染程度的多项指标均表现出一定的相关性，如水体不ＢＯＤ５、无机氮、ＣＯＤ、无机磷等的含量及采样点离陆源排放点的距离等相关性较大，其相关性在水动力条件差的水域尤为显著。     

Nereis diversicolor: an Unreliable Biomonitor of Metal Contamination in the 'Ría de Bilbao' (Spain)

Abstract. Samples of the polychaete Nereis diversicolor O.F. M üller and surficial sediments from a clean and a polluted estuary were taken during a year at monthly intervals to determine, by comparison, the range of temporal fluctuations in trace metal concentrations. Whereas metal variations in surficial sediments were more pronounced in the polluted estuary, concentrations in N. diversicolor in both estuaries showed a wide range of temporal variation that surpassed that of surficial sediments. A clear decline in all metals studied (except Mn) from polluted sediments was noted after episodes of strong rainfall. In the reference estuary the fluctuations were less pronounced. Metal body burdens in N. diversicolor from the two estuaries were usually quite similar even though total levels in sediments differed considerably, indicating a lack of worm-sediment relationship. The low level of Cu, Cr and Pb in the tissues of N. diversicolor suggest a mechanism which prevents the polychaete from being exposed to these metals in highly polluted sediments. Therefore, it is concluded that N. diversicolor cannot be considered to be an ideal biomonitor of metal contamination in the polluted Bilbao Estuary.     

Cadmium-binding proteins in the blue crab, callinectes sapidus: laboratory-field comparison

The blue crab, Callinectes sapidus, is distributed along the east coast of the United States from Cape Cod, Mass., through the Gulf of Mexico, including both relatively unpolluted coastal areas and estuaries contaminated with trace metals. Cadmium is of particular concern because it is concentrated in the digestive glands of blue crabs and can be passed on to consumer organisms. Tissue concentrations and partitioning of trace metals from crabs exposed in the laboratory to 10 ppb dissolved cadmium for 40 days were compared with blue crabs collected from two locations on the Hudson River, NY, Foundry Cove and Haverstraw Bay, Foundry Cove and Haverstraw Bay, both of which have elevated trace metal levels relative to estuarine areas near Beaufort, NC. Crab digestive glands, gills and muscle were removed and analyzed for total cadmium, copper, zinc and nickel concentrations using acid digestion and atomic absorption spectrophotometry, and metal-binding (metallothionein-like) proteins were determined by gel-filtration chromatography. In crabs exposed to cadmium in the laboratory, the cytosolic partitioning was similar to previous investigations at our laboratory where higher levels of cadmium (100 ppb) and shorter exposure times (14 days) were used. The similarity in cadmium partitioning from these two separate experiments indicates dose independence. In crabs from polluted environments the digestive glands contained the highest concentrations of trace metals. Chromatograms of the cytosol from the digestive glands and gills from both field and laboratory exposed crabs showed similar distributions of cadmium, copper and zinc. The gills of both field and laboratory-exposed crabs had metal-binding proteins that contained mostly cadmium, and the digestive glands had metallothionein-like proteins that contained cadmium, copper and zinc. Estimated molecular weights for these proteins were similar to the metallothioneins found in other crustaceans and mammals.     

Effect of pollution on genetic diversity in the bay mussel Mytilus galloprovincialis and the acorn barnacle Balanus glandula.

To test if environmental contamination acts as a selection force affecting genetic diversity at the population level, two intertidal invertebrate species, Mytilus galloprovincialis and Balanus glandula, were collected from seven different bay sites in southern California. Collections were made at three relatively pristine 'clean' sites and four 'impacted' sites exposed to heavy industrial or boating activity, and which had previously been identified as having measurable levels of pollution. Genetic diversity at each site was assessed by comparing fragment polymorphisms generated from genomic DNA by randomly amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). All populations retained a large amount of genetic diversity and were genetically similar to each other. However, several different measures of diversity indicated that, for most primers, the populations of both species from impacted sites had lower genetic diversity compared to those populations from clean sites. Individuals at impacted sites were more likely to share the same haplotypes than were those from clean sites. Few bands seen in the clean sites were absent from the impacted sites or vice versa, but a number of bands in the clean site populations were significantly less common in the impacted populations, while a few bands uncommon in clean site populations were more common at impacted sites. Together, these results suggest that pollution at the impacted sites may reduce genetic diversity among the resident invertebrate populations.     

Oxidative stress responses in two populations of Laeonereis acuta (Polychaeta, Nereididae) after acute and chronic exposure to copper

Acute (4 days) and chronic (14 days) effects of copper were evaluated on the antioxidant defenses of Laeonereis acuta (Polychaeta) collected in unpolluted (UP) and polluted (P) sites. In the acute assay (125 and 250 micro g Cu/l) superoxide dismutase (SOD) and glutathione S-transferase (GST) activities did not change, whereas catalase (CAT) increased in worms from both the sites. Lipid peroxidation was higher in copper exposed worms from the P site. In the chronic assay (62.5 micro g Cu/l) polychaetes from the P site showed enhanced activities of SOD, GST and CAT and higher contents of metallothionein-like proteins and sulfhydrils compared to worms from UP. Differences in responses between polychaetes from UP and P sites suggest that organisms from the polluted site, P, are more susceptible to oxidative stress conditions.     

Relationship between heavy metal accumulation and genetic variability decrease in the intertidal crab Pachygrapsus marmoratus (Decapoda; Grapsidae)

The “genetic erosion” hypothesis posits that heavy metal stress is related to a loss of genetic diversity at the population level. The genetic diversity of natural populations can, however, be affected by natural processes as well as by human impact. We studied the relationship between heavy metal bioaccumulation and genetic variability in the intertidal crab Pachygrapsus marmoratus. Tissue samples were collected from 40 individuals inhabiting four polluted and four unpolluted sites along the Tuscan coast (Mediterranean basin), and were examined for four heavy metals (arsenic, As, cadmium, Cd, lead, Pb, and copper, Cu). We also assessed the genetic variability of 235 crabs from the same localities using six microsatellite loci.Our results show that the bioaccumulation levels of these individuals accurately reflect the levels of pollution in their immediate environment, and that heavy metals accumulate more in the hepatopancreas than in the gills. Moreover, populations from polluted sites have significantly less genetic variability, measured as mean standardized d², and a significantly lower percentage of unrelated individuals, than populations from unpolluted sites. This evidence supports the “genetic erosion” hypothesis for metal heavy exposure in natural environments.     

Trace metal assessment in sediment, molluscs and reed leaves in the bay of follonica (Italy)

Marine surficial sediments, intertidal molluscs and reed leaves collected in the Bay of Follonica are slightly contaminated by Pb, Ni, Zn and Fe. The main pollution sources are the waste water and the atmospheric emission of industrial plants. Probably, the weathering of the pyrite deposits along the eastern coast of Elba Island and adsorption and coprecipitation with hydrous Fe-Mn oxides increase the trace metal concentrations in sediments from the Piombino Channel. Mussels and limpets are contaminated by Hg because of the higher background level of the northern Tyrrhenian Sea.     

The Influence of Primary Production Rate on the Population Dynamics of Patella granulans, an Intertidal Limpet

Abstract. The production rates of intertidal algae at seven study sites in the Saklanha Bay area. South Africa, were measured monthly for 15 months. Algae grew rapidly and formed extensive, permanent mats on shores that were enriched with nutrients from the dissolved guano of seabirds. The most significant correlate of limpet (Patella granulans) growth was the rate of algal production; more rapid limpet growth on nutrient-rich shores, combined with the effects of predation by African Black Oystercatchers Haematopus moquini on limpets, led to differences in the size structures and life-history patterns of populations of limpets on enriched and unenricheci shores. The rate of primary production affected relationships between intertidal organisms at all trophic levels and influenced patterns of organization within the community.     

Microbial diversity in polluted harbor sediments II: Sulfate-reducing bacterial community assessment using terminal restriction fragment length polymorphism and clone library of dsrAB gene

Sulfate-reducing bacteria (SRB) are important regulators of a variety of processes in coastal marine sediments regarding organic matter turnover, biodegradation of pollutants, and sulfur and carbon cycles. Yet their community compositions have not been investigated in polluted harbor sediments. This study described the diversity and spatial variation of SRB communities in surface sediments in Victoria Harbor, Hong Kong. The spatial variation of SRB communities was described by terminal restriction fragment length polymorphism (T-RFLP). The results showed that the most diversified terminal restriction fragments were found at polluted sites. In addition, cluster analysis indicated that although the SRB communities were different at the two polluted sites, they were still more similar to each other than to the two more distant reference sites. Based on a dsrAB clone library constructed at a polluted site, diversified SRB were found, represented by 30 Operational Taxonomic Units (OTUs). Upon comparisons among the SRB sequences detected from this study and those in the GenBank, five clades of SRB were found. Three clades belonged to the known families Desulfobacteraceae, Desulfobulbaceae, and Syntrophobacteriaceae. The majority of sequenced clones, which distantly related to sequences in the GenBank, constituted the remaining two unclassified groups, suggesting unique SRB members related to the polluted harbor environment. Statistical analyses indicated that estimated SRB richness correlated with environment factors such as sulfur content, acid volatile sulfate, and redox potential.     

Using benthic macrofauna to assess environmental quality of four intertidal mudflats in Hong Kong and Shenzhen Coast

Abstract-Intertidal zone is a significant wetland between land and ocean. It plays an important rolein maintaining local ecological balance. Both Mai Po and Futian intertidal mudflats are located in Shen-zhen Bay and are important "refueling" point along the East Asian/Australian flyway of migratory birds.The environmental quality of Mai Po and Futian mudflats have aroused great concern due to rapid eco-nomic developments in Hong Kong and Shenzhen in recent decades. Microinfauna of Mai Po and Futianmudflats was investigated in December 2000 and the faunal data were used to assess their environmentalquality. Two other mudflats, namely Ma Wan Typhoon Shelter (a more disturbed area) and Luk Kengintertidal mudflat (a place with relatively less human disturbance), were also sampled for macroinfaunaand used as reference mudflats. Shannon-Weaver species diversity index (I_(sd)), biotic coefficient (C_b)and macrofaunal pollution index (I_(mp)) of the macrofauna community on four intertidal mudflats wereused to     

Molluscan lysosomal responses as a diagnostic tool for the detection of a pollution gradient in Tolo Harbour, Hong Kong

Sub-cellular perturbations in the lysosomal compartment of molluscan haemocytes were examined in mussels (Perna viridis), collected along a pollution gradient. The neutral red technique was validated using a well defined contamination gradient among indigenous populations from five stations along Tolo Harbour, Hong Kong. Condition indices (shell length:dry tissue wt) and tissue metal concentrations were also measured in an attempt to identify a relationship between contamination level and adverse physiological effects. Correlations were found between lysosomal retention time and condition along the pollution gradient. There were significant differences between mussels collected from stations on offshore islands and those collected from inner harbour sites (p < 0.05). There was, however, little correlation between metal concentrations and retention time or condition (p > 0.05).     

Preliminary results of a field study on some stress-related parameters in Tapes philippinarum naturally infected by the protozoan Perkinsus sp.

The aim of the present work was to investigate the possible use of the clam Tapes philippinarum—a filter-feeding bivalve living on soft bottoms—in the monitoring of sediment pollution in the Lagoon of Venice (North-East Italy). To this end, two clam populations, exposed to different environmental conditions (a relative clean site and an industrial polluted area), have been monitored for the prevalence of the protozoan parasite Perkinsus sp. as well as for several biological parameters commonly used as determinants of contaminants exposure. First results showed a low prevalence, higher in the clean site, but a very diffuse distribution of the parasite throughout the two populations. Nevertheless, greater digestive cells height and longer lysosomal latency time were observed in clams from the clean area. As for the physiological stress-related parameters, even if clearance rates were always higher in these clams, scope for growth values indicated a more complicated biological response.     

Limpets as prey: Lipophrys pholis choice for size

Patellid limpets are key species on rocky shores, structuring intertidal assemblages through their grazing. Their role as prey for fish is, however, often overlooked in studies of intertidal ecosystem functioning. The shanny Lipophrys pholis is a common predator of limpets on rocky shores in Northern Europe and, in this study, we examined the characteristics of its predation on limpets based on prey size. The limpet size range that adult L. pholis (10–12.5 cm) can eat was examined in the laboratory using Patella depressa in 0.2 cm classes, varying between 0.5 cm and 3.0 cm in maximum shell length. There was a limpet size refuge above 1.8 cm, while all smaller sizes were readily consumed by the shanny. The predator attacking behaviour was also examined and found to vary with prey size. Limpets up to 0.8 cm were crushed by the jaw‐constricting force and eaten whole, whereas larger prey were prised from the substratum and the body subsequently separated from the shell. To examine whether there was a L. pholis preference for P. depressa size, a two‐stage laboratory experiment was done using two size classes defined as small (0.7–0.8 cm maximum shell length) and large (1.5–1.6 cm maximum shell length). In the first stage, the predator was given each limpet size class separately. In the second stage, the fish was given a choice between the two classes. Lipophrys pholis was shown to have a preference for the large size class (1.5–1.6 cm). The average number of limpets consumed by the shanny was examined for the duration of one high‐tide typical of the central region of Portugal (≈ 4 h). On average, approximately five limpets (up to an eight limpet maximum) were consumed. The present study shows that L. pholis has a feeding preference based on limpet size, which suggests that the limpet population structure and intra‐ and inter‐specific interactions may be influenced by the shanny predation.     

Evaluating environmental contamination in Ria Formosa (Portugal) using stress indexes of Spartina maritima.

Species of Spartina are potentially useful for biomonitoring coastal systems because they are abundant in the intertidal and have a wide geographic distribution in temperate zones. In this study, three indexes of physiological stress, thiolic protein concentration, adenylic energy charge (AEC) index [AEC = ATP + 0.5ADP/(ATP + ADP + AMP)] and photosynthetic efficiency (Fv/Fm), are measured in Spartina maritima plants collected at four sites and compared with metal concentrations in the sediments and Spartina tissues. Two sites were close to urban centers and two were located further away, in less contaminated areas. Concentrations of copper, cadmium, lead and zinc were determined by atomic absorption spectrophotometry, thiolic proteins were determined by polarography and adenilic nucleotides were determined by high performance liquid chromatography. Photosynthetic efficiencies were measured with a fluorometer. The sediments of sites closer to urban centers had higher concentrations of metals than the sites located further away. Metal concentration within plants did not follow this spatial pattern, probably due to spatial differences in metal bioavailability, uptake rates, internal transport and excretion processes. Concentrations of all metals studied were higher in underground plant structures than aerial, except cadmium whose levels were not significantly different. On the other hand, the stress indexes followed the spatial pattern of metals in the sediment. Thiolic protein concentrations were higher in plants from polluted sites, and were three to four times higher in aerial than in underground plant structures. Leaf AEC ratios and photosynthetic efficiencies were lower in plants from polluted sites indicating that they were growing under stressful conditions. We conclude that the use of these indexes in S. maritima represent a useful tool to monitor contamination in Ria Formosa lagoon as they provide an integrated measure of the toxicological burden of contamination.     

Effect of seasonality on oxidative stress responses and metal accumulation in soft tissues of Aulacomya atra,a mussel from the South Atlantic Patagonian coast

This study investigated the effects of pollution and its interaction with temperature on the oxidative status of the ribbed mussel Aulacomya atra in the southern Atlantic Patagonian coast. Animals were collected from four sites with different degree and type of human activity impact, during the summer and winter of 2011. Seawater chromium, copper, manganese, nickel and zinc concentrations were measured, as well as metal accumulation, lipid peroxidation, protein oxidation, reduced glutathione levels, and enzymatic activities of superoxide dismutase and glutathione-S-transferase in gills and digestive glands.Metal bioaccumulation and oxidative stress responses in both tissues were generally higher in mussels from harbor areas. Water temperature had a remarkable effect on gill SOD activity and protein oxidation during winter in mussels from all locations.Methodologically, we conclude that measuring both metal bioaccumulation and oxidative stress responses allowed for a more accurate assessment of the biological effects of metal present in seawater.     

Trace metal content in sediments and autochthonous intertidal organisms from two adjacent bays near Ushuaia,Beagle Channel (Argentina)

The aim of this work was to monitor levels of Cd, Cu, Pb, Zn and Fe in sediments, mussels (Mytilus edulis chilensis) and limpets (Nacella magellanica) from the Industrial zone (IZ); fuel dock (FD) and Ushuaia Peninsula (UP) on the Beagle Channel. In sediments, seasonal variations showed high values of Cu and Pb in spring and Zn in autumn. Comparing among sites, Cd concentration was superior in UP (2.07 μg/g); while Pb was maximum in FD (41.00 μg/g). In mussels, a higher bioaccumulation in winter was found. Mussels from UP showed the highest bioaccumulation of Cu (5.95 μg/g) and those from FD presented the highest of Zn (170.15 μg/g). A seasonal trend was not found for limpets, while differences among sites were observed for Cd being the highest at IZ (3.02 μg/g). Although pollution level found was low, anthropic activities at the studied sites could result in deterioration, further monitoring is recommended.     

南极菲尔德斯半岛潮间带南极帽贝的种群生态学研究──夏季种群数量变化和垂直分布

于1994年12月—1995年3月（南极夏季）期间,对南极菲尔德斯半岛卵石滩、岩石滩潮间带南极帽贝的种群数量、垂直分布进行了定点、定时、定量的调查和观察。结果表明,南极帽贝的种群数量有明显下降,分别仅为起始数量的35％和52％。卵石滩潮间带的帽贝分布于中潮区及低潮区,尤其是低潮区,且有随季节（冬季来临）向潮下带移动的趋势。岩石滩潮间带的帽贝具有类似的垂直分布特征,但其在分布带上的发生频次有很大差异,呈波动状态。在垂直岩面上,帽贝数量在低潮区的底部更为集中。讨论了导致数量变化、垂直分布的可能原因。今后有必要深入研究种群数量的时空动态及其调控机理。     

Assessment of environmental quality and inland water pollution using biomarker responses in caged carp (Cyprinus carpio): Use of a bioactivation:detoxication ratio as a Biotransformation Index (BTI)

In the present study the bioaccumulation of poly chlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) as well as the responses of a suite of biochemical parameters were investigated in a standardized carp (Cyprinus carpio) fish line. Carps were caged for 2 to 8 weeks at two Amsterdam freshwater sites: the relatively unpolluted Outer IJ and the heavily polluted Volgermeerpolder. The primary objective of the experiments was to develop a reliable and reproducible method to monitor the inland water pollution and to classify the environmental quality of freshwater sites. The highest tissue levels of organic trace pollutants were generally found in carp that was caged for at least four weeks at the polluted site. Hepatic phase I enzymes showed the highest pollution-induced responses, while phase II enzymes and serum transaminases were less sensitive. The antioxidant enzymes showed virtually no response to pollutants. In order to assess effects on the balance between bioactivation and detoxication the ratio between phase I (EROD) and phase II (GST or UDPGT) activities was introduced as a biotransformation index (BTI). Highest BTI values were observed in carp that was caged for 4 to 6 weeks at the polluted site. The BTI values decreased after eight weeks of exposure due to elevated phase II activities.     

NADPH-diaphorase activity in the central nervous system of the Gray mussel Crenomytilus grayanus (Dunker) under stress conditions: a histochemical study

NADPH-diaphorase (NADPH-d) is a histochemical marker for nitric oxide synthase (NOS) and is widely used to identify nitric oxide (NO) producing cells in the central nervous system (CNS) of both vertebrates and invertebrates. NADPH-d histochemistry was used to quantitatively characterize putative NO-producing neurons in the CNS of the Gray mussel Crenomytilus grayanus subjected to two kinds of stress, environmental pollution and hypoxia, the latter caused by the mollusk transportation in a small volume of water. Mussels were sampled from one relatively clean (reference) and four polluted sites in Amursky and Ussuriysky Bays (Peter the Great Bay, Sea of Japan) in August, 2003. The number of NADPH-d-positive neurons was estimated and enzyme activity was determined from the optical density of the formazan precipitate in the CNS ganglia at 0, 3, and 72 h after sampling. Just after sampling, NADPH-d-positive neurons were found in the cerebropleural, visceral, and pedal ganglia. The number and staining intensity of NADPH-d-positive neurons were significantly higher in the pedal ganglia than the other two ganglia. There were significant differences in the number of NADPH-d-positive neurons and enzyme activity between the mussels from the reference and heavily polluted stations. The proportion and staining intensity of NADPH-d-positive neurons were maximum in the pedal ganglia of the mussels from the heavily polluted station in Amursky Bay. Transportation of mussels in a limited volume of water for 3h resulted in a significant increase in the proportion and staining intensity of NADPH-d-positive neurons in all ganglia. In mollusks from all stations kept in aerated aquaria for 72 h, both the proportion and staining intensity of NADPH-d-positive neurons did not differ significantly from the initial level. However, the differences in the proportion and staining intensity of NADPH-d-positive neurons between the reference and heavily polluted stations were significant. The present results suggest that NO is involved in mollusk nerve cell adaptation to environmental changes.