Effects of molting and environmental factors on trace metal body-burdens and hemocyanin concentrations in the American lobster, Homarus americanus

Hemocyanin concentrations in the hemolymph of marine crustacea are dependent on the molt cycle and on environmental conditions. Studies in our laboratories have found that hemocyanin levels in blue crabs are reduced after ecdysis and under conditions of environmental stress (Engel, Brouwer, & McKenna, 1993. Hemocyanin concentrations in marine crustaceans as a function of environmental conditions. Marine Ecology Progress Series, 93, 233-244). We have extended those studies to include the American lobster, Homarus americanus. Hemolymph and digestive gland tissues from Long Island Sound lobsters were analyzed for hemocyanin, copper, and zinc during different stages of the molt cycle. Hemocyanin, copper and zinc in the hemolymph were highest in premolt stages (D1-D4), and lowest in the postecdysal papershell stages (B1-B2). Concomitantly, copper in digestive glands decreased significantly following ecdysis, but no significant changes in the metals bound to metallothionein (MT) were observed. Copper-MT was the predominant form throughout the molt cycle, presumably because lobsters were obtained from copper-contaminated areas. To examine the effects of environmental factors, intermolt lobsters were collected from locations of different environmental quality along the Atlantic coast, and were analyzed for hemocyanin and trace metals. In general, animals from areas with a history of contamination showed the highest hemocyanin concentrations.     

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.     

Morphological colour change in the American lobster (Homarus americanus) in response to background colour and UV light

American lobster (Homarus americanus) have a thick calcified cuticle, and do not exhibit rapid colour changes characteristic of other crustaceans. Thus, the plasticity of their coloration has been largely overlooked. Colour in lobsters is determined by the amount, location, and form of the carotenoid pigment astaxanthin, and it is possible for lobsters to alter colour by changing one of these characteristics of astaxanthin deposition. Here, short‐term colour variation in American lobster in response to environmental cues (background colour and ultraviolet (UV) light) was investigated in a laboratory experiment. Lobsters were reared in conditions controlling background colour (white, black) and UV light (present, absent). Digital photographic analysis was used to determine how these conditions influenced the luminescence (light or dark) of lobster colour, as well as the ratio of red to blue hues. Of the environmental variables considered within this experiment, UV light was the predominant factor, and caused lobsters to become darker in colour. In the absence of UV light, lobsters matched background colour, and turned darker in response to the darker background. Environmental matching has practical implications both for wild lobsters as they settle to the benthic habitat, and for enhancement programmes, to grow lobsters that are best suited for local habitats.     

Metallothionein-like proteins induced by cadmium stress in the scallop Mizuhopecten yessoensis

Organisms have evolved a cellular response called stress protein response that increases their tolerance in adverse environmental conditions. Well known stress proteins that bind essential and toxic metals are metallothionein (MT). The scallop Mizuhopecten yessoensis is the most interesting organism because it is able to accumulate toxic cadmium in its digestive gland. However, in the tissue of the digestive gland of Mizuhopecten yessoensis MT (metallothioneins) have not been found. Eastern scallops, Mizuhopecten yessoensis, were collected from two locations ?? one clean and one polluted site. The concentrations of cadmium (Cd), copper (Cu) and zinc (Zn) were measured in the digestive gland. There was a significant increase in Cd concentrations in this studied tissue. We found that in the presence of cadmium Mizuhopecten yessoensis can induce high molecular proteins. The results of experiments have shown that Cd-binding ligands have a number of properties similar to MT: acetone and temperature stability; the ability to bind some metals, including Cd, Cu and Zn. Protein chromatography (FPLC, Superosa 12) from the digestive gland of scallop M. yessoensis has shown that cadmium is associated with high molecular weight Cd-binding proteins (72 kDa and 43 kDa). The major cadmium-binding protein 72 kDa is glycoprotein. In experiments we have demonstrated that Cd-binding proteins can be induced when there is cadmium exposure. The results of this study strongly suggest that the far eastern scallop Mizuhopecten yessoensis has a unique and well-developed system for the detoxification of heavy metals and it allows for biochemical systems to be maintained in a relatively stable manner in the presence of heavy metals.     

Altered glutathione status in atlantic croaker (Micropogonias undulatus) tissues exposed to lead

Metals which induce the synthesis of metallothioneins, such as cadmium and mercury, also elevate glutathione (GSH) concentrations in fish tissues.^1,2 To determine whether metals which do not stimulate metallothionein synthesis increase GSH levels, the GSH status of Atlantic croaker tissues was examined after oral administration of lead. Lead significantly increased GSH levels in the intestine and liver, but not in kidney or brain tissues. Lead was not bound to GSH and the increase in GSH did not appear to be linked to lead accumulation in these tissues. Investigations of hepatic GSH synthesis and turnover in vitro using a radiolabelled amino acid precursor, C¹⁴ glycine, showed that the elevation of GSH was due to an increase in GSH synthesis rather than decreased utilization of GSH. These studies suggest that the interactions between GSH and lead are indirect and involve a stimulatory effect of the metal on the activity of the enzymes for GSH synthesis.     

Bioaccumulation des métaux chez le mytilidae hydrothermal Bathymodiolus sp. de la ride médio-atlantique

The hydrothermal environment is characterised, among other things, by a high metal concentration, related to the convective sea-water circulation inside the oceanic crust and its interaction with basaltic rocks. The biological communities associated with the hydrothermal system can survive in this toxic environment owing to their ability to regulate their intracellular metal levels by excretion or accumulation of metal ions in non-toxic forms. Various detoxification processes have been previously studied within an hydrothermal organism: immobilisation and precipitation of the metal in lysosomal systems, or binding to specific and soluble ligands such as metallothioneins. The later are heat stable and characterised by a high content of cysteine. The quantification of Fe, Zn, Cu, Mn, Cd, Ag, Ba and Sr in the tissues of the vent mussel Bathymodiolus sp. indicates a high accumulation of these metals in two target organs, the gill and the digestive gland. This accumulation is in relation with high concentrations of metallothioneins, but the subcellular distribution of metals indicates a higher contribution of the insoluble compartment for detoxification processes. These results are compared with the literature data about metal bioaccumlation in Bathymodiolus thermophilus collected at the Galapagos Rift.     

Organic matter production of American lobsters (Homarus americanus) during impoundment in Maine,United States

Lobster impoundments are dammed coastal embayments utilised to hold American lobsters (Homarus americanus) before shipping to market. The impacts of lobster impoundments on the environment have not been previously studied. Here, the digestive functioning of American lobsters was examined to assess the quantity and quality (TVS, total volatile solids) of lobster faeces produced under the temperatures and feeding regimens these animals were subjected to during the impoundment period. Overall it was determined that quantity and quality of faeces did not differ among the experimental temperatures (5, 10, and 15°C), and that animals fed every 1–2 days produced greater quantity and quality of faeces than those fed every 3–18 days, or those animals fed less than every 18 days. As a first estimate of organic matter production in active lobster impoundments, it was calculated that a typical lobster impoundment produced 0.79 ± 0.35 (average ± 1 SD) g TVS m^‐2 day^‐1 during the impoundment period, equivalent to 3.11 ± 0.60% of the total weight of lobsters stocked into the impoundment. This level of organic matter production is below the level produced by other aquaculture operations, and that at which benthic impacts might be expected.     

An improved electrochemical method for the quantification of metallothioneins in marine organisms

A six-to nine-fold increase in detection limits of the Brdic?ka polarographic assay for metallothioneins has been achieved through the employment of a commercially available static mercury drop electrode (SMDE) and reduced temperatures. At 16°C detectable concentrations of metallothionein (MT-1) from the crab Scylla serrata were in the range of 5 to 10 μg litre^-1. Operating conditions and electrode preparation are discussed. For a number of thiolic molecules a linear relationship between polarographic response and either the number of cysteinyl residues or the molecular weight was noted. In the latter case on enhancement of response by sulfur-metal bonds is suggested. The utility of this assay permits a rapid quantitative assay for these proteins at basal levels in marine invertebrates and other organisms.     

Partitioning of metals among metal-binding proteins in the bay mussel, Mytilus edulis

Metabolism of metals was assessed in populations of Mytilus edulis exposed to metals in the laboratory and in the field. Chronic exposure to metals in both environments resulted in increased concentrations of metals in both the low molecular weight metal-binding protein fraction, which contains metallothioneins, and in the high molecular weight metal-binding protein fraction, which contains metalloenzymes. Our results from analysis of laboratory-exposed populations and from monitoring indigenous and transplanted populations indicate that the capacity for production of metallothioneins is limited and that the quantities present differ greatly with seasonal changes in the environment.     

Eelgrass (Zostera marina L.) as an indicator organism of trace metals in the Limfjord,Denmark

The concentrations of Pb, Cu, Cd and Zn were determined in above- and belowground parts of eelgrass (Zostera marina L.) at forty stations in a shallow, brackish water area (the Limfjord, Denmark). The concentrations of the trace metals were significantly elevated near the cities of Aalborg (Pb, Cu) and Struer (Cd). Trace metal concentrations in above and belowground parts of eelgrass were log-normal distributed and the concentrations of Cd, Cu and Zn in aboveground parts were significantly higher than in belowground parts. Furthermore, a significant correlation between trace metal concentrations in above- and belowground parts was found. The background level of trace metal concentrations in eelgrass in the Limfjord was estimated.The application of eelgrass as a monitoring organism is discussed; it is suggested that the concentration of some trace metals in above- and belowground parts of eelgrass may be used as a measure of the bioavailable fraction of these trace metals in ambient and interstitial water (sediment), respectively.     

Effect of reproductive state and sex on movement and food consumption of western rock lobster (Panulirus cygnus) in a tank environment

The catchability of western rock lobsters (Panulirus cygnus) during the spawning season (October and November) may be affected by their reproductive state. In this study, movement and food consumption of males and females of different reproductive states held in laboratory tanks were measured as proxies for catchability. Density of lobsters was found to influence both movement and food consumption. Solitary animals and, to a lesser extent, two lobsters per tank, were more sedentary and consumed significantly less food than animals at a density of three or four specimens per tank. Tests using three animals per tank confirmed that mature, unmated, and ovigerous females carrying early stage eggs moved more frequently away from their shelters than males or females carrying late stage eggs (P < 0.0001, d.f. = 3). Further, unmated females and females with early stage eggs recorded significantly higher food consumption (P < 0.0061, d.f. = 3) compared with males or females carrying late stage eggs. The greater time spent away from shelter and also food consumption make unmated females and females with early stage eggs likely to be more catchable than males or females with late stage eggs. As catchability is commonly used to obtain population estimates from survey data, these results have implications for surveys which use catch rates of breeding animals as indicators of egg production.     

Accumulation and effects of organotin compounds in oysters and mussels: Correlation with serum biochemical and cytological factors and tissue burdens

Oysters and mussels exposed to a concentration of 0·7 ppb (μg/liter) tributyltin from painted panels in flowing seawater accumulated tin in the digestive glands to comparable levels. The mussels experienced approximately 50% mortality during the 60-day test period, but the oysters suffered virtually no deaths. There was no evidence from either bivalve of elevated numbers of hemocytes during the test period and no evidence for cellular disruption as detected by increased levels of serum lysosomal hydrolases. Serum protein of exposed mussels relative to controls increased with time of exposure to the toxicant, while oyster serum protein, normally 10 x higher than in mussels, did not. No evidence was found for elevated stress proteins (heat shock proteins) or metallothioneins in the serum hemocytes of either bivalve. Responses by these animals to fatal or near fatal doses of TBT were thus very different from responses to copper that we have reported elsewhere.^1,2     

罗纳河中悬浮物质的痕量元素的研究

于1989年5月－1990年4月间,在罗纳河下游近河口处进行每月一次为期间个月的样品采集工作,并且样品用HNO3－HF－HCIO4湿法消化后,用原子吸收分光光度法测定了悬浮颗粒物中的Co,Cr,Cu,Mn和Pb含量。结果表明,在不同月份之间悬浮物中的痕量元素组成具有显著地差别,其中一些元素的浓度随流量的增加而减少,而另外一些似乎与流量之间没有显著的关联。在稳定体系的假设下,所估计的人文活动对痕量元素的贡献可占观测浓度的10％－50％。当与其它河流对比时,发现罗纳河中的颗粒态痕量元素的浓度明显高于那些受人文活动影响较小的体系,而与北美的河流系统接近,表明罗纳河流域的人文活动对河流中颗粒态痕量元素的影响是显著的。     

Evaluating local population dynamics of the American lobster,Homarus americanus,with trap‐based mark‐recapture methods and seabed mapping

Seabed mapping, spatially referenced trapping, and mark‐recapture methods have all been useful tools in ecological studies of lobsters and other benthic animals. Here we integrate the three methods to evaluate local population dynamics and movements of American lobsters, Homarus americanus, in coastal fishing grounds in Maine, United States. The study was conducted on five study areas of different size, and used two different sampling protocols. At one site (1 km² in area) we used a monthly mark‐recapture sampling interval over a 6‐month period, only tagging a subsample of the catch. At four smaller sites (0.3 km²) we used a shorter‐term approach, sampling at 3–4‐day intervals for a 2‐week duration, tagging the entire catch. Tagging data were analysed with a modified Jolly‐Seber model adapted for continuous sampling to estimate population abundance, gains (immigration), and losses (emigration and mortality). Side‐scan sonar surveys of the seabed combined with diver‐based population surveys, stratified by substrate type, provided an independent comparison to mark‐recapture‐based estimates of abundance over the same areas. Spatial referencing of trap catch also allowed us to relate catch rates and lobster movements directly to seabed features. The longer‐term tagging data on the larger study area provided abundance estimates that were more consistent with the diver observations, and estimates of gains and losses statistically more robust, than those derived from the shorter‐term effort on the smaller sites. The flux of lobsters followed the well known seasonal movements on these fishing grounds, with gains and losses from the larger study area ranging over 1000 individuals per day, and an estimated mid‐summer peak density of >65 000 lobsters per km² (individuals >50 mm carapace length). This approach may lend itself to broader application with the American lobster.     

Bioavailability of heavy metals bound to sediments affected by a mining spill using Solea senegalensis and Scrobicularia plana

After the Aznalcóllar mining spill (April, 1998) different ecosystems along the Guadiamar river and the Guadalquivir estuary were impacted by high concentrations of metals. The concentration of metals Zn, Cd, Pb, Cu and As has been detected as a good tracer of the pathway of the toxic spill. Different individuals of the fish Solea senegalensis and the clam Scrobicularia plana were exposed to contaminated sediments after the accidental spill in the south-west Spain. The induction of metallothioneins in fish and the bioaccumulation of metals in clams were determined in different tissues to determine the biological adverse effect associated with the heavy metals from the spill. A relationship between the enrichment of metals in the tissues and the induction of metallothioneins was determined for those assays performed using contaminated sediments. The bioaccumulation of metals indicates that the risk associated with the spill should be continuously evaluated to establish the sublethal effects related to the accident.     

Assessment of potential biomarkers, metallothionein and vitellogenin mRNA expressions in various chemically exposed benthic Chironomus riparius larvae

The objective of this study was conducted to identify the possibility of using Chironomus metallothionein (MT) and vitellogenin (VTG) as biomarkers of stress caused by endocrinedisrupting chemicals (EDCs), heavy metals, herbicides and veterinary antibiotics. We characterized the MT and VTG cDNA in Chironomus riparius and evaluated their mRNA expression profiles following exposure to different environmental pollutants. The gene expression analysis showed that the MT mRNA levels increased significantly after long-term exposure to cadmium (Cd), copper (Cu), Lead (Pb), di(2-ethylhexyl) phthalate (DEHP), and 2,4-dichlorophenoxyacetic acid (2,4-D). Moreover, the VTG mRNA expression increased significantly in C. riparius larvae exposed to BPA, NP, DEHP, Cd, 2,4-D and fenbendazole. Evaluation of the long-term effects of environmental pollutants revealed up regulation of Chironomus MT mRNA in response to DEHP exposure among EDCs, and the level of the VTG mRNA was increased significantly following treatment with Cd and herbicide 2,4-D at all concentrations in a dose-dependent manner. These results indicate that VTG could be used as a potential biomarker of herbicide and Cd as well as EDCs, while MT was a potential biomarker of heavy metals such as Cd, Cu, and Pb in aquatic environments.     

Trace metals in bivalve molluscs from Thailand

Capture fisheries and aquaculture provide a large quantity of bivalve molluscs for human consumption in Thailand each year. The existing information on trace elements in these bivalves is fragmentary and contradictory. This paper reports data for eight metals (cadmium, chromium, copper, iron, lead, nickel, mercury and zinc) in the four major species marketed. These are the clam Pahia undulata, the cockle Anadara granosa, the green mussel Perna viridis and the rock oyster Crassostrea commercialis. The bivalves analysed were taken at markets or in the field to cover as many of the major sources as possible. The results indicate no major threat to public health from trace elements in these species from Thailand and lend confidence to the expansion of aquaculture operations. In particular, the levels of lead found in the samples analysed are considerably lower than those previously reported by other authors.     

Dietary change of the rock lobster Jasus lalandii after an ‘invasive’ geographic shift: Effects of size,density and food availability

During the 1990s the rock lobster Jasus lalandii shifted its focus of distribution south-eastwards along the coast of South Africa, to establish a dense population in an area where it was previously rare. This coincided with a marked decrease in the sea urchin Parechinus angulosus, a preferred prey item of J. lalandii and a vital source of shelter for juveniles of the abalone Haliotis midae. The range expansion of lobsters has thus economic and ecological ripple effects. We determined the diets of small (50–65 mm carapace length) and large (>69 mm CL) rock lobsters from gut content analyses, and compared them between three ‘lobster invaded’ sites and three adjacent ‘non-invaded’ sites where densities are still low. At the non-invaded sites, diets were collectively heterogeneous but the dietary breadth of individual lobsters was narrow (in contradiction to generally accepted ecological theory), and the lobsters fed mainly on large, individual, mobile, high-energy prey such as sea urchins and large winkles. Conversely, at invaded sites where lobster densities were high, they consumed predominantly small, colonial or sessile low-energy prey such as sponges, barnacles and foliar algae, and the diet was significantly more uniform among individuals, but broader within individuals. This was a direct result of the contrasting benthic community structure of the two areas, and consequent prey availability – itself caused by differences in intensity of rock-lobster predation. Cannibalism was unexpectedly greater at non-invaded sites, possibly as a result of lobsters being larger there. The diet of small and large lobsters also differed significantly. Large rock lobsters predominantly consumed large individual prey such as lobsters, urchins and crabs, while small rock lobsters ate mainly colonial, sessile prey such as sponges and barnacles, and small prey such as tiny winkles and crustaceans. Dietary selectivity indices revealed that algae and sponges were negatively selected (avoided) in non-invaded areas but positively or neutrally selected in invaded areas. These dietary differences have important ramifications not only for the lobster populations but also for the structure and functioning of the radically different communities that have developed in invaded areas, reflecting a regime shift induced by lobster predation.     

Etude du transfert de quelques oligo-éléments métalliques entre le milieu sédimentaire estuarien et les poissons plats ‘mangeurs de sédiments’

In order to study the transfer of trace metals (Cd, Pb, Cu, Zn) between sediments and living organisms, flatfishes (flounder, Platichthys flesus and sole, Solea solea) were collected monthly in the Loire estuary (France) for more than a year. Trace metal concentrations were determined in flatfish digestive contents and tissues. The influence of some ecological factors upon bioaccumulation of trace metals was considered.We consider the benthic way of life not to be responsible for a preferential bioaccumulation of trace metals in flatfishes. Although zinc accumulation in external organs is important, the hypothesis of an uptake via food and sediment cannot be rejected.     

Accumulation and cytosolic partitioning of metals in the American oyster Crassostrea virginica

The ability of the oyster to accumulate trace metals, particularly the transition metals (Cd, Cu, Zn, and Hg), from food and water has been well documented. There is little information, however, on how these metals interact to affect whole animal retention, partitioning, and binding to metallothionein (Mt). In this study oysters were exposed to Cd and Cu, both alone and in combination, and significant effects have been demonstrated on subcellular partitioning. These studies showed that Cu can displace Cd from Mt, but the Cd is not lost from the animal. Two metal-binding protein peaks have been separated by gel chromatography (<10 K and 24 K daltons). The lower molecular weight protein peak has been characterized as Mt, consisting of two isoproteins, but the higher molecular weight protein peak appears to be an aggregation of metal-binding proteins, some of which may be Mt or a Mt dimer. In environments that are contaminated with trace metals, Mt binds primarily Hg, Cu, and/or Cd and Zn, but in noncontaminated areas Mt binds primarily Zn on one of the two isoMts.