Population differences in nerve resistance to paralytic shellfish toxins in softshell clam, <Emphasis Type="Italic">Mya arenaria</Emphasis>, associated with sodium channel mutations

The softshell clam, Mya arenaria, is a commercially important bivalve with wide latitudinal distribution in North America. Populations of clams with a history of repeated exposure to toxic Alexandrium spp. have developed a natural resistance to the paralytic shellfish toxins (PSTs) produced by these algae. An association between PST resistance in individual clams and a single mutation in the saxitoxin (STX) binding region of the α-subunit of the voltage-gated sodium (Na⁺) channel gene was previously identified. Here we establish that more than one mutation associated with nerve resistance to STX occurred at this locus. Both cDNA from mRNA and genomic DNA sequences from individual clams are identical demonstrating that both alleles are expressed simultaneously. In addition, one resistant allele per individual is sufficient to confer neural resistance to STX even though heterozygous individuals show an intermediate level of resistance to STX in in vitro nerve trunk assays.     

Paralytic shellfish toxins sequestered by bivalves as a defense against siphon-nipping fish

Saxidomus giganteus (butter clams), are known to sequester diet-derived paralytic shellfish toxins (PST), highly potent neurotoxins, in their siphons. Captive staghorn sculpins (Leptocotus armatus), a marine fish species known to crop bivalve siphons, developed a significant aversion to siphons from toxic but not non-toxicS. giganteus following a single conditioning feeding of toxic siphon tissues. Control fish showed no aversive response to siphons from non-toxicS. giganteus during 11 feeding sessions over 56 d. Aversive and non-aversive behavior varied with the toxicity of the siphons, but not with the geographic origin of the clams. Both experimental and control fish ate freely and showed no aversion to siphons from toxic littleneck clams (Protothaca staminea). Littleneck clams, unlikeS. giganteus, retain PST in their visceral mass but not in their siphons. Both toxic and non-toxicS. giganteus extended their siphons significantly more often and higher above the sediment surface during dark hours, but toxicS. giganteus extended their siphons higher than non-toxic individuals. These results support the hypothesis that siphon-nipping by fish may have selected for the retention of PST in butter clam siphons as a chemical defense.     

Toxic dinoflagellates (Alexandrium fundyense and A. catenella) have minimal apparent effects on oyster hemocytes

The possible effect of Alexandrium spp. containing paralytic shellfish poisoning (PSP) toxins on the hemocytes of oysters was tested experimentally. In one trial, eastern oysters, Crassostrea virginica Gmelin, were exposed to bloom concentrations of the sympatric dinoflagellate, Alexandrium fundyense Balech, alone and in a mixture with a non-toxic diatom, Thalassiosira weissflogii (Grun) Fryxell et Hasle. Subsequently, another experiment exposed Pacific oysters, Crassostrea gigas Thunberg, to a mixed suspension of the sympatric, toxic species Alexandrium catenella (Whedon et Kofoid) Balech, with T. weissflogii. Measurements of numbers of oyster hemocytes, percentages of different cell types, and functions (phagocytosis, reactive oxygen species (ROS) production, and mortality) were made using flow-cytometry. During and after exposure, almost no significant effects of Alexandrium spp. upon hemocyte numbers, morphology, or functions were detected, despite observations of adductor-muscle paralysis in C. virginica and measured toxin accumulation in C. gigas. The only significant correlation found was between toxin accumulation at one temperature and higher numbers of circulating live and dead hemocytes in C. gigas. The PSP toxins are known to interfere specifically with sodium-channel function; therefore, the finding that the toxins had no effect on measured hemocyte functions suggests that sodium-channel physiology is not important in these hemocyte functions. Finally, because oysters were exposed to the living algae, not purified toxins, there was no evidence of bioactive compounds other than PSP toxins affecting hemocytes in the two species of Alexandrium studied.     

Feeding and absorption of the toxic dinoflagellate Alexandrium tamarense by two marine bivalves from the South China Sea

Paralytic shellfish poisoning (PSP) toxins can be accumulated by bivalves through the feeding process; therefore, knowledge on feeding and the assimilation of PSP-toxin-containing algae is critical to understand the kinetics of PSP toxins in these bivalves. In the South China Sea, it has been documented that the scallop Chlamys nobilis has a much higher PSP toxin burden than the clam Ruditapes philippinarum. Experiments were therefore carried out to assess whether the difference in toxin burden between these two species of bivalves was due to differences in feeding and absorption. In a mixed diet of Alexandrium tamarense (a PSP-toxin-producing dinoflagellate) and Thalassiosira pseudonana (a non-toxic diatom), the maximum clearance and filtration rates were about two times higher in the scallop C. nobilis than in the clam R. philippinarum. Furthermore, the clams produced pseudofeces at a lower cell density than the scallops. However, we found that the clams were unable to selectively exclude the toxic dinoflagellates by pseudofeces production. The scallop C. nobilis also possessed a greater ability to assimilate A. tamarense with a comparable carbon absorption efficiency to the diatom T. pseudonana. In contrast, the carbon absorption in the clam R. philippinarum was lower when feeding on A. tamarense than on the diatom. In general, the absorption efficiency decreased with increasing concentration of A. tamarense. Thus, it is likely that the higher PSP toxin levels in the scallops compared with clams can be partly explained by differences in their feeding and absorption behavior. Other processes, especially the biotransformation and biokinetics of PSP toxins, may also play a significant role in defining the inter-species differences in PSP body burden in marine bivalves.     

Differential effects of Aureococcus anophagefferens isolates ("brown tide") in unialgal and mixed suspensions on bivalve feeding

Picoplanktonic brown tides of Aureococcus anophagefferens have had devastating effects on production of commercially exploited bivalve populations in shallow, mid-Atlantic estuaries in the United States. The toxin produced by this alga has not been chemically characterized. This study develops a bioassay using juvenile mussels, Mytilus edulis, based on the inhibitory effect of brown tide on bivalve suspension-feeding, to compare the cellular toxicity of three Long Island, New York, clonal isolates of A. anophagefferens. Two recent (1995) isolates (CCMP 1707 and 1708) from Peconic Bay proved highly toxic and caused greater than 100-fold reduction in clearance rates (CR) of juvenile mussels in unialgal and mixed suspensions with a nutritious alga, Isochrysis galbana (clone T-iso), relative to controls. A third 1986 isolate from Great South Bay (CCMP 1784) showed no detectable toxicity in 24-h trials, and may have lost its initial potency over more than a decade of laboratory culture. Identification of a non-toxic strain provides a useful tool for future research. Cultures of the toxic isolate CCMP 1708 in late-stationary growth phase were significantly more toxic than those in early-exponential phase. The threshold concentration of toxic A. anophagefferens cells that inhibits clearance on co-occurring phytoplankton species was determined for juvenile (10-mm) hard clams, Mercenaria mercenaria. Relatively low concentrations (⣃᎒³ to 50᎒³ cells ml^-1) of isolate CCMP 1708 were sufficient to sharply reduce clam CR of I. galbana. Calculations based on these results suggest that, at peak historical densities of M. mercenaria in Great South Bay, removal of A. anophagefferens at low cell densities by suspension-feeding benthos could provide an effective top-down grazing control mechanism to prevent the initiation of brown tide in shallow, inner bays.     

Dinoflagellate <Emphasis Type="Italic">Alexandrium leei</Emphasis> (Dinophyceae) from Singapore coastal waters produces a water-soluble ichthyotoxin

A strain of Alexandrium leei Balech that was isolated in October 2002 from Singapore coastal waters and identified by light and scanning electron microscopy and phylogenetic analysis using LSU rDNA sequences, is toxic to Asian sea bass fingerlings (Lates calcarifer Bloch). The ichthyotoxicity of the algal cells obtained by filtration (15 μm mesh net) and rinsed with sterile culture medium indicated that the toxicity of A. leei was probably not due to bacterial contamination, which was further supported by a negative correlation between the time to death of the fish and the dosage of algal cells applied. Paralytic shellfish toxins (PST) could not be detected indicating that PST was not the cause of fish mortality. Fish bioassays using frozen culture, heat-treated cultures, cell-free culture medium, and hexane, ethyl acetate, and water extracts of algal cells indicated that A. leei produces a heat-stable, polar ichthyotoxin(s) which can be released from the algal cells into the culture medium. Phylogenetic analysis based on the LSU rDNA sequences of A. leei confirmed its identification and indicated that the Singapore strain is more similar to isolates from Malaysia than to a geographically distant strain from Korea. This is the first evidence of icthyotoxin production by A. leei.     

Effects of two paralytic shellfish toxin producing dinoflagellates on the pelagic harpacticoid copepod Euterpina acutifrons

The effects of two paralytic shellfish toxin (PST) producing dinoflagellates, Alexandrium minutum Halim (high and low toxin strains) and Gymnodinium catenatum Graham, on the pelagic harpacticoid copepod Euterpina acutifrons Dana were tested in a series of experiments run from October 1994 to May 1995. In small volumes (350 ml), both strains of A. minutum (300 to 350 cells ml^-1), and G. catenatum (175 cells ml^-1), strongly reduced naupliar activity (about 30 and 17% were inactive after 24 h, respectively). Activity is here defined as movement. In medium volumes (6 litre), 40% of nauplii incubated with the high toxin strain of A. minutum (1000 cells ml^-1) and 8% of nauplii incubated with cell-free filtrate of the same culture were inactive after 24 h; these values increased to 50 and 30% respectively after 3 d. In large volumes (20 litre), adult copepods incubated with A. minutum (1000 and 10000 cells ml^-1) for 5 d revealed only trace levels of PSP-toxins (paralytic shellfish poisoning) in the extracts analysed by HPLC. With both strains of A. minutum (1000 and 10000 cells ml^-1), 10 to 15% of the copepods were inactive after 1 to 2 d. It is suggested that E. acutifrons avoids feeding on the dinoflagellates after tasting a few cells, but that the dinoflagellates may exude toxins or other substances that affect the copepods. The inactivating effect of the toxic dinoflagellates on the nauplii was more rapid and stronger than on adult copepods, although strong inactivation and death were also observed in adults with time (up to 80% were inactive after 5 d of incubation with A. minutum). Still, in our experiments a considerable proportion of adult females incubated with the toxic dinoflagellates remained active and were able to produce viable eggs for several days.     

Sea otter foraging on deep-burrowing bivalves in a California coastal lagoon

Sea otter, Enhydra lutris, predation had no detectable effect on abundance and size distribution of deep-burrowing bivalve prey in the Elkhorn Slough, California, USA. Up to 23 otters were present for 6 mo of the study period (March 1984 through April 1985). This is in contrast to previous studies of sea otter predation, especially on the shallow-burrowing Pismo clam Tivela stultorum, which can be found along the wave-exposed coast near the slough. The deep-burrowing clams Tresus nuttallii and Saxidomus nuttalli made up 61% of the prey taken in the slough, and are more difficult for otters to excavate than Pismo clams. The occurrence of foraging otters was highest in an area where the two bivalve prey were extremely abundant (18 individuals m^–2). However, the otters did not selectively prey on the largest clams available within the study sight, but foraged preferentially in a patch of smaller individuals where bivalve burrow depth was restricted by the presence of a dense clay layer. This foraging strategy maximized the amount of prey biomass obtained per unit volume of sediment excavated. Our findings suggest that in soft-sediment habitats deep-burrowing bivalves may be more resistant to otter predation than shallower burrowers.     

Cytological study and immunohistochemical location of PSP toxins in foot skin of the ormer, Haliotis tuberculata, from the Galician coast (NW Spain)

The ormer, Haliotis tuberculata Linnaeus, 1758, is able to accumulate paralytic shellfish toxins (PST) in the foot side skin. Epidermal and secretory cells are described separately for the pedal side and sole epithelia, as notable differences have been observed. Melanin and photosynthetic pigment granules in the epidermal cells of the foot side epithelium give this skin its appearance and characteristic color. This epidermis also has abundant secretory cells containing at least three kinds of secretory granules. Conversely, the epidermal cells of foot sole epithelium do not have pigment granules, and the secretory cells are mostly subepithelial. Immunohistochemical reaction against PST (saxitoxin and its derivatives) showed that the toxins are located in specific cells of the foot side epithelium, which are distinct from the epidermal cells. Immunofluorescence also reveals toxins throughout many areas of the mucilage covering the epithelium, suggesting the secretory nature of the cells containing the toxins. These results do not show any apparent relationship between melanin or photosynthetic pigment granules and PST content. PST were not detected in the foot sole epithelium either immunohistochemically or by HPLC. Considering the toxic characteristics of ormers described in previous studies and the results presented here, the authors discuss the role of toxins retained or produced by the foot side epithelium Received: 25 July 2000 / Accepted: 7 November 2000     

UV-absorbing substances in zooxanthellate and azooxanthellate clams

The effects of UV-A and UV-B radiation on photosynthesis of zooxanthellae within the siphonal mantle of the giant clam, Tridacna crocea, and in isolation were studied. While UV-B irradiation (2.4 W m⁻², 20 min) completely suppressed photosynthesis of the isolated zooxanthellae, it had little effect on their photosynthetic ability if they were irradiated while within the siphonal mantle of the host tissue. Chemical analysis of the siphonal mantle of T. crocea showed the presence of significant amounts of mycosporine-like amino acids (MAAs), which absorb UV-A and -B light. However, no MAA was detected in the isolated zooxanthellae. MAAs were concentrated in the siphonal mantle and kidney tissues in comparison with other tissues. In the siphonal mantle, MAA concentrations were the highest in the outermost surface layer where most of the zooxanthella cells resided. This indicates that the zooxanthellae are protected from UV radiation by a screen of concentrated MAAs in the host clam. Aside from T. crocea, significant amounts of MAAs were found not only in other zooxanthellate clams, such as T. derasa, Hippopus hippopus, Colculum cardissa and Fragum unedo, but also in a closely related azooxanthellate clam, Vasticardium subrugosum. On the other hand, no MAA was detected in any of the zooxanthellae from these zooxanthellate clams. No MAA was detected in the tissues of a deep-sea bivalve, Calyptogena soyoae. Although MAAs seem to block strong UV radiation in the shallow-water clam, they are probably not essential for the clam's life in the dark. MAAs in shallow-water clams may be derived from food and accumulated in their tissues, especially in the siphonal mantle and kidney. Received: 29 November 1996 / Accepted: 13 January 1997     

Transfer of brevetoxins to a tellinid bivalve by suspension- and deposit-feeding and its implications for clay mitigation of <Emphasis Type="Italic">Karenia brevis</Emphasis> blooms

Blooms of the brevetoxin-producing Karenia brevis in the Gulf of Mexico cause massive fish kills, food poisoning and adverse respiratory effects in humans. Sedimentation of toxic cells following inert clay application could reduce toxin incorporation by commercially important suspension-feeding bivalves and thus prevent direct public health impacts, but could potentially lead to brevetoxin (PbTx) accumulation by benthic deposit-feeders. The goal of this study was therefore to compare suspension- and deposit-feeding as pathways for brevetoxins. We investigated: (1) the effect of toxic K. brevis on both feeding modes using a facultative deposit-suspension feeding tellinid bivalve, the clam Macoma balthica, as a model species and (2) the relative effectiveness of brevetoxin transfer via suspension- and deposit-feeding over 24-h exposure. Sedimentation of K. brevis was achieved by treatment with 0.25 g phosphatic clay l⁻¹ and brevetoxin concentrations were measured by ELISA. Karenia brevis reduced both suspension- and deposit-feeding activity. This study demonstrates that brevetoxins can be rapidly accumulated by a surface deposit-feeding bivalve from sedimented K. brevis cells and that comparable toxin levels can be attained by both suspension- and deposit-feeding modes [1.2–1.6 μg PbTx (g tissue wet weight)⁻¹]. Deposit-feeding clams generally do not pose a direct threat to humans but may provide a pathway for brevetoxin food web transfer.     

Size-dependent photosynthetic performance in the giant clam <Emphasis Type="Italic">Tridacna maxima</Emphasis>, a mixotrophic marine bivalve

Giant clams form a symbiosis with photosynthetic algae of the genus Symbiodinium that reside in clam mantle tissue. The allometry of symbiont photosynthetic performance was investigated as a mechanism for the increasing percentage of giant clam carbon respiratory requirements provided by symbionts as clam size increases. Chlorophyll fluorescence measurements of symbionts of the giant clam Tridacna maxima were measured during experiments conducted in September of 2009 using specimens 0.5–200 g tissue wet weight (3–25 cm long), collected from waters around southern Taiwan (N 21°36′, E 120°47′) from July to August of 2009. Light-dependent decreases in effective quantum yield (∆F/F _m′) calculated as the noontime maximum excitation pressure over PSII (Q _m), relative electron transport rates (rETR), and dark-adapted maximum quantum yield (F _v/F _m) all varied as a quadratic function of clam size. Both Q _m and rETR increased as clam size increased up to ~10–50 g then decreased as clam size increased. F _v/F _m decreased as clam size increased up to ~5–50 g then increased as clam size increased. Chlorophyll fluorescence measurements of rETR were positively correlated with gross primary production measured during chamber incubations. Overall, symbionts of mid-sized clams ~5–50 g exhibited the highest light-dependent decreases in effective photosynthetic efficiencies, the highest relative electron transport rates, and the lowest maximum photosynthetic efficiencies, and symbiont photosynthetic performance is allometric with respect to host clam size.     

Toxin accumulation and feeding behaviour of the planktonic copepod Calanus finmarchicus exposed to the red-tide dinoflagellate Alexandrium excavatum

The planktonic copepod Calanus finmarchicus is a dominant member of the zooplankton community in the lower St. Lawrence Estuary in eastern Canada. Blooms of the toxic marine dinoflagellate Alexandrium excavatum which produces high cellular levels of paralytic shellfish poisoning (PSP) toxins, occur during the period of high C. finmarchicus production in summer in this region. To study the feeding behaviour of C. finmarchicus in the presence of Alexandrium spp., experiments were conducted in which female adult copepods collected from the St. Lawrence Estuary between May and September 1991 were exposed under controlled conditions to two toxic isolates of A. excavatum (Pr18b and Pr11f) from the estuary and to a non-toxic control (PLY 173) of a closely related species, A. tamarense isolated from the Tamar Estuary, Plymouth, U.K. Clearance rates on non-toxic A. tamarense cells averaged 5.5 ml ind^-1 h^-1 but were nearzero with either toxic isolate. When presented with a mixture of A. excavatum and the non-toxic diatom Thalassiosira weissflogii in varying proportions, C. finmarchicus fed upon the diatom but avoided the toxic dinoflagellate. Although feeding rates on A. excavatum were very low, toxin analysis by high-performance liquid chromatography with fluorescence detection (HPLC-FD) revealed that the PSP toxins were accumulated in copepods exposed to toxigenic dinoflagellates.The toxin composition in copepods was similar to that of the toxic dinoflagellate, but not necessarily identical, particularly after short-term (2-h) exposure, when relatively elevated levels of N-sulfocarbamoyl toxins were detected. The evidence suggests that C. finmarchicus ingests toxic dinoflagellate cells, either mistakenly or during exploratory bouts of feeding, and accumulates PSP toxins in its gut system and perhaps in other tissues.     

Effects of salinity on the clam <Emphasis Type="Italic">Chamelea gallina</Emphasis>. Part I: alterations in immune responses

In the present study, the effects of differing salinities on some important functional responses of haemocytes from the clam, Chamelea gallina, were investigated. The animals were kept for 7 days at 28‰ (hyposalinity), 34‰ (control) and 40‰ salinity (hypersalinity), and total haemocyte count (THC), haemocyte volume, phagocytosis, lysozyme-like activity (in both haemocyte lysate and cell-free haemolymph) were measured. The survival-in-air test was also performed. Clams kept at 28‰ showed significantly increased THC with respect to animals kept at 34 and 40‰. The analysis of haemocyte size frequency distribution highlighted that in clams kept at 28‰ the haemocyte fraction of about 5 μm in diameter and 50–100 femtolitre in volume increased markedly. Conversely, in animals kept at 40‰ an increase was observed in the haemocyte fraction having about 8–10 μm diameter and 400–500 femtolitre volume. Higher phagocytic activity was recorded in haemocytes from control clams, with respect to that of clams kept at 28 and 34‰. Lysozyme-like activity in haemocyte lysate was shown to increase significantly in animals kept at 28‰ with respect to that of clams kept at 40‰, whereas enzyme activity in cell-free haemolymph from clams kept at 34‰ was significantly higher with respect to that of clams maintained at 40‰. A relationship between phagocytosis and lysozyme secretion is suggested. The resistance to air exposure of clams kept at 28 and 40‰ was shown to decrease significantly; LT₅₀ values fell from 7 days in clams kept at 34‰ to 4 and 5 days in those kept at 28 and 40‰, respectively. Results demonstrated that salinity values far from 34‰ affects the functional responses of haemocytes and reduce the resistance of clams to exposure to air.     

Very slow development in two Antarctic bivalve molluscs,the infaunal clam <Emphasis Type="Italic">Laternula elliptica</Emphasis> and the scallop <Emphasis Type="Italic">Adamussium colbecki</Emphasis>

Embryos of the large infaunal clam Laternula elliptica and the scallop Adamussium colbecki, from Antarctica, were cultured over an 18-month period. Their development rates were extremely slow, taking 240 and 177 h, respectively, to reach the trochophore stage. This is ×4 to ×18 slower than related clams and scallops from temperate latitudes. The relationship between temperature and development rate for bivalve molluscs shows the expected slowing with reduced temperature (Q ₁₀ in the range 2–4) for temperate and tropical species. However, the slowing at polar latitudes is much stronger than at warmer waters, and all of the limited data for Antarctic species are well above the Arrhenius plot for the overall bivalve data, and the Q ₁₀ value for Antarctic to cool temperate species is 11.8, well outside the expected range for biological systems. Either the relationships describing the effects of temperature on the kinetics of biological systems do not apply to Antarctic bivalve molluscs, or some other factor that cannot be compensated for becomes important at low temperature. In the laboratory, L. elliptica embryos stayed viable in very sticky egg capsules for up to 18 months without hatching. However, even the disturbance of removing eggs using a pipette ruptured some egg capsules allowing embryo release. Gametogenesis in Antarctic marine invertebrates is almost universally slowed compared to temperate species, with nearly all cases documented requiring more than 1 year to complete oogenesis. The only exception so far appears to be A. colbecki, which has a 1-year gametogenic cycle. The data here indicate that it has been unable to adapt embryonic development in a similar way, and we are not aware of any exceptions to the markedly slowed development at low temperature rule.     

Comparative study of the carbon cycle in Venus verrucosa fed on bacteria and phytoplankton

Following a previous study, this paper deals with the utilisation of a phytoplankton suspension (Pavlova lutheri) by the clam Venus verrucosa Linné, 1767 as a function of time. Experimental clams filtered and ingested 100% of the suspension after 2 h. The radioactivity recovered in the whole soft body of the clams was 83.8% after 2 h, 66.0% after 10 h and 46.0% after 41 h. The faeces contained 13.7% after 3 h and 34.3% after 41 h. Carbon-dioxide radioactivity (gas plus dissolved) increased slowly to 12.4% after 41 h. When the water was not changed after 17 h, the clams reabsorbed a great part of their dissolved products and faeces. Under these latter conditions the radioactivity recovered from the whole soft body of the clams was roughly the same as that recovered after the start of the experiment. The aim of this study was the comparison of the consumption of two kinds of food: bacteria and phytoplankton. The work was carried out over three years, from 1977 to 1980.     

Field depuration and biotransformation of paralytic shellfish toxins in scallop<Emphasis Type="Italic"> Chlamys nobilis</Emphasis> and green-lipped mussel<Emphasis Type="Italic"> Perna viridis</Emphasis>

Under laboratory conditions, the scallop Chlamys nobilis and the mussel Perna viridis were exposed to N-sulfocarbamoyl toxins (C2 toxin), a paralytic shellfish toxin (PST), by feeding a local toxic strain of the dinoflagellate Alexandrium tamarense (ATDP) that produced C2 toxin exclusively. The bivalves were subsequently depurated in the field, and their depuration kinetics, biotransformation and toxin distribution were quantified. Depuration was characterized by a rapid loss within the first day, followed by a secondary slower loss of toxins. In the fast depuration phase, scallops detoxified PSTs more quickly than the mussels (depuration rate constants for scallops and mussels were 1.16 day^–1 and 0.87 day^–1, respectively). In contrast, the mussels detoxified PSTs more quickly than the scallops in the slow depuration phase, and the calculated depuration rate constants (mean+SE) from day 2 to day 13 were 0.063+0.009 day^–1 and 0.040+0.019 day^–1 for mussels and scallops, respectively. The differences in the appearances of gonyautoxins, GTX2 and GTX3, and their decarbamoyl derivatives, dcGTX2, dcGTX3 and GTX5, which are all derivatives of C2 toxin, indicated active and species-specific biotransformation of the algal toxins in the two bivalves. In both species of bivalves, the non-viscera tissue contained fewer toxins and lower concentrations than the viscera-containing tissue compartment. In scallops, very little toxin was distributed in the adductor muscle. In mussels, most of the PSTs were found in the digestive gland with significant transport of toxins into the digestive gland from other tissues during the course of depuration. The toxin profiles of scallops and mussels differed from each other and from that of the toxic algae fed. A significant fraction of GTX5 was detected in the mussels but not in the scallops. Our study demonstrates a species specificity in the depuration kinetics, biotransformation and tissue distribution of PSTs among different bivalves.Communicated by T. Ikeda, Hakodate     

The effects of the diarrhetic shellfish poisoning toxins,okadaic acid and dinophysistoxin-1, on the growth of microalgae

The diarrhetic shellfish poisoning toxins okadaic acid (OA) and dinophysistoxin-1 (DTX-1) are potent phosphatase inhibitors produced by certain species of marine dinoflagellates. OA can cause hyperphosphorylation of a broad range of animal and higherpalnt proteins, but little is known regarding the effects of the DSP toxins on marine organisms or their biological function. A variety of microalgae, including a clone ofProrocentrum lima known to produce both OA and DTX-1, were incubated with solutions of OA and in one case DTX-1 or a combination of OA and DTX-1. OA inhibited the growth of all non-DSP-producing test species at micromolar concentrations, butP. lima was not affected even at much higher levels. This differential activity of OA suggests that the DSP toxins may play an allelopathic role and raises questions regarding the strategies adopted by DSP-producing dinoflagellates such asP. lima to avoid autotoxicity. The effects of DTX-1 on microalgal growth were found to be equivalentt to those of OA, and the effects of both toxins in combination were simply additive.     

Differences in sterol composition of clams (<Emphasis Type="Italic">Ruditapes decussatus</Emphasis>) from three rías in Galicia (NW Spain)

The sterol composition of three different populations of Ruditapes decussatus from three localities close to each other, but where different environment conditions prevail, was investigated over a period of 14 months. Sterols of adult clams were isolated by thin layer chromatography and identified by gas chromatography/mass spectrometry. In all samples, the major sterol component was cholesterol (>40% of total sterols); other sterols identified were 24-norcholesta-5,22-dienol, 22-cis-dehydrocholesterol, 22-trans-dehydrocholesterol, brassicasterol, 24-methylene-cholesterol, campesterol, stigmasterol, -sitosterol and isofucosterol. At each locality, changes in sterol levels followed specific and different patterns, which remained constant for the period studied and allowed the origin of the clams to be distinguished. Stepwise discriminant analysis, based on the percentage amounts of the sterols at each locality, indicated brassicasterol, 22-cis-dehydrocholesterol, cholesterol, 24-norcholestadienol and -sitosterol as discriminant variables that distinguish clam populations. Correct identification of the clam origin was achieved in 100% of the samples, demonstrating that sterols can justifiably be used as molecular biomarkers for determining the origin of this bivalve species.Communicated by O. Kinne, Oldendorf/Luhe     

Ecology of the bivalve-inhabiting hydroid Eugymnanthea inquilina in the coastal sounds of Taranto (Ionian Sea,SE Italy)

The life history of the bivalve-inhabiting hydroid Eugymnanthea inquilina was investigated in two different hosts, the mussel Mytilus galloprovincialis (collected from 1989–1991 from the Ionian Sea, Italy) and the clam Ruditapes decussatus (collected from 1991–1992, Ionian Sea, Italy). Hydroids living in mussels revealed a size selection for hosts longer than 40 mm, being almost completely absent in mussels below this size. This might be controlled by signals linked to the sexual maturity of the bivalve. The proportion of molluscs inhabited showed a seasonal trend in mussels only, with a sharp decline at the onset of medusoid liberation. Production of medusoids was high in mussels, whereas medusoids were rare and often abortive in clams. This indicates a higher degree of fitness of E. inquilina in mussels than in clams. The hydroid life cycle was not completed in clams, which therefore were presumably supplied by planulae from medusoids produced by mussel-inhabiting hydroids. Mussels played a key role in the stability and persistence of E. inquilina populations in the studied area. Selective ingestion of trematode sporocysts by E. inquilina hydroids indicated a protective role of the hydroid against mussel parasites, leading to reconsideration of this symbiotic association as a possible mutualism rather than a simple inquilinism.