Response of Asterias vulgaris to chemical stimuli

Groups of the sea star Asterias vulgaris approached a variety of chemical stimuli in a flow tank. Urea, lactic acid, succinic acid, and handsweat were attractive to some animals; some simple acids and amino-acid mixtures were weakly attractive; pH changes and sea-water salts were not attractive. Although the groups were similarly maintained and responded with high internal consistency, they differed in responsiveness to different substances. These variations are not easily accounted for as contamination or effects of prior animal experience. A. vulgaris detects many simple organic substances, but the behavioral response is not constant and species-wide; it depends on nonchemical factors. This sensory/behavioral mode presumably suits a nonspecialist predator, and may be widespread among similar asteroids. Individuals attracted to lactic acid could be induced to cling to and evert their stomachs over a sponge soaked in lactic-acid solutions. The chemicals found atrractive in this study do not account for most of the attractiveness of shellfish tissue extracts studied previously.Contribution No. 2854 of the Woods Hole Oceanographic Institution.     

Effects of a phycotoxin,okadaic acid,on oyster heart cell survival

Okadaic acid (OA) is a dinoflagellate toxin which accumulates in shellfish producing diarrhetic shellfish poisoning (DSP) in humans. It was found that OA is a highly selective inhibitor of protein phosphatase types 1 (PP1) and 2A (PP2A) which produces a marked increase in phosphorylation of several proteins, including p38 mitogen-activated protein (MAP) kinase. The cytotoxicity attributed to OA and the effects on p38 MAP kinase and calcium current were examined in the oyster Crassostrea gigas in this study. Data showed that p38 MAP kinase is strongly expressed in oyster heart and that OA bioaccumulated in cultured heart cells. Hence the effects of OA was tested in vitro and in vivo on oysters. OA was found to (i) exert a positive chronotropic effect on cultured atrial cardiomyocytes which is related to an increase in calcium current via PKC as shown by patch clamp measurements, (ii) produce an activation/phosphorylation of MAP kinase as shown by Westernblot while the non-phosphorylated p38 remained constant during treatment, (iii) did not induce a pro-apoptotic effect. Data suggest that OA may also stimulate the anti-apoptotic pathway by phosphatase inhibition.     

Effects of the dinoflagellates Karlodinium veneficum and Prorocentrum minimum on early life history stages of the eastern oyster (Crassostrea virginica)

The bloom-forming dinoflagellates Prorocentrum minimum and Karlodinium veneficum can have detrimental effects on some marine life, including shellfish, but little is known about their effects on early life history stages of bivalves. In the Chesapeake Bay region, blooms of these dinoflagellates overlap with the spawning season of the eastern oyster, Crassostrea virginica. In laboratory experiments, we compared the effects of P. minimum and K. veneficum on the survival and development of embryos and larvae of the eastern oyster. At 10⁴ cells ml⁻¹, P. minimum did not have a negative effect on embryos and larvae in 2-day exposures. The yield of D-hinge larvae was equal to or greater than in control treatments. At 2 × 10⁴ cells ml⁻¹ (approximately equal biomass to the P. minimum treatment) K. veneficum caused significant mortality to oyster embryos within 1 day and almost no embryos developed into D-hinge larvae. This effect was not alleviated by the provision of an alternate food source (Isochrysis sp.). Significant mortality was observed when larvae were exposed to K. veneficum at concentrations of 10⁴ cells ml⁻¹ (approximately 5 ng ml⁻¹ of karlotoxin). The K. veneficum cultures used in these experiments were relatively low in toxin content, more toxic strains could be expected to cause mortality at lower cell concentrations. Survival and maturation of embryos and larvae may be reduced when spawns of the eastern oyster coincide with high bloom densities of K. veneficum.     

Molecular prey identification in wild Octopus vulgaris paralarvae

The trophic ecology of Octopus vulgaris paralarvae collected in 2008 off the Ría de Vigo, NW Spain (42° 12.80′ N–9° 00.00′ W), was approached by both morphological and molecular methods. External digestion of prey and posterior suction of the liquefied contents by wild O. vulgaris paralarvae made the morphological identification of gut contents impossible. Thus, a PCR-based method using group-specific primers was selected to identify prey consumed by O. vulgaris paralarvae in the pelagic realm. The mitochondrial ribosomal 16S gene region was chosen for designing group-specific primers, which targeted a broad range of crustaceans and fishes but avoided the amplification of predator DNA. These primers successfully amplified DNA of prey by using a semi-nested PCR-based approach and posterior cloning. Homology search and phylogenetic analysis were then conducted with the 20 different operational taxonomic units obtained to identify the putative organisms ingested. The phylogenetic analysis clustered ingested prey into 12 families of crustaceans (11 belonging to the order Decapoda and 1 to the order Euphausiacea) and two families of fishes (Gobiidae and Carangidae). According to the Czekanowski’s Index (CI), the trophic niche breadth of O. vulgaris paralarvae is low (CI = 0.13), which means that these paralarvae are specialist predators at least during the first weeks of their life cycle. It is the first time that natural prey has been identified in O. vulgaris paralarvae collected from the wild, and such knowledge may be critical to increasing the survival of O. vulgaris hatchlings in captivity, a goal that has been actively pursued since the 1960s by aquaculture researchers.     

Uptake and accumulation of naphthalene by the oyster Ostrea edulis,in a flow-through system

A flow-through system was used to follow naphthalene and naphthalene metabolite accumulation in the seawater and in the tissue of the oyster Ostrea edulis. After 72 h, 82.5% of the naphthalene carbon was recovered from the system. Glucose was added to seawater to stimulate the pathways of glucose metabolism in the oysters. Streptomycin (100 ppm) reduced microbial oxidation of naphthalene and glucose, and reduced bacterial growth. However, even in the presence of streptomycin, microbial oxidation of naphthalene was considerable. The main oxidation product recovered from seawater was ¹⁴CO₂. Radioactivity was also associated with compounds which separated by TLC with 2- and 1- naphthol. The pattern of naphthalene uptake and accumulation in oyster tissues was relatively constant after only a few hours of exposure to naphthalene. The potential of tissues to accumulate naphthalene was shown to be a function of multiple variables such as nutritional state, lipid concentration, length of exposure to naphthalene, and the external naphthalene concentration. Carbon-14-labeled metabolites derived from ¹⁴C-naphthalene were consistently recovered from digests of the oyster tissues. Non-CO₂ alkaline-soluble substances were the primary metabolites. Hexane-extractable substances, which separated by TLC with known standards of 2- and 1- naphthol, were consistently recovered from seawater and tissue digests. It was not possible to conclude that these metabolites were a result of naphthalene metabolism by oyster enzyme systems.     

Metal Concentration in Oyster,Crassostrea Gigas,and Sediment in Ann-Ping Mariculture Ground,Taiwan

Abstract

This study was to assess the metal contamination in oyster tissue grown in the Ann-ping mariculture ground in Taiwan. the information generated from this work also revealed general metal pollution problem for Taiwan's oyster farmers. Oysters, Crassostrea gigas, and surficial sediments collected from ten locations in Ann-ping mariculture ground in Taiwan for metals concentration (Cu, Zn, Pd, Cd, Fe and Mn) were performed. Analytical results indicated that the yearly averaged oyster copper concentrations (μg g^?1, wet weight) in oyster soft parts from Ann-ping increased from 21.3±4.1 in 1993; 24.1±6.8 in 1994; 36.8±11.9 in 1995 to 43.9±23.1 μg g^?1, wet weight, in the 1996 raising season. the mean oyster copper concentration reached a level of 50 μg g^?1, wet weight, in December 1996. This increasing trend of metal concentration in oyster tissue indicates a potential pollution source which may pose a potential disaster as green oyster incidence, which occurred on the Charting coast in 1986, in Taiwan. Sediment samples in Ann-ping mariculture ground were also collected and examined. the seasonal variation of the copper concentration in surficial sediment from Ann-ping did not show an increasing trend as observed in oyster tissue.     

Allozyme variation in European populations of the oyster Ostrea edulis

Variations at 22 enzyme coding loci were surveyed in 11 populations of the oyster Ostrea edulis L., which were sampled between 1988 and 1990 along the Atlantic and Mediterranean coasts of Europe. Atlantic oyster beds suffered a steady decline during the last century, and restocking of beds with oysters of foreign origin has probably resulted in a high degree of interbreeding of natural oyster stocks from all Atlantic Europe. Our study confirms the low levels of genetic variability previously reported for the oyster populations from the Atlantic coasts, and extends it to the Mediterranean coasts. The locus arginine-kinase (ARK ^*) exhibited a high degree of interpopulation differentiation (F _ST=0.289), resulting from extensive variation in gene frequencies along a geographical cline. However, the overall genetic differentiation between populations was slight, and similar to that reported for other local populations of bivalves (mean genetic distance between populations is 0.010, mean F _ST=0.062). A general pattern of increasing differentiation along the coastline in an Atlantic-mediterranean direction emerged; but genetic differentiation among the Atlantic populations was not significantly lower than that observed among the Mediterranean populations. This and other results suggest that the effects of extensive transplantation of oysters among various areas in Europe are detectable only in some particular localities. The geographical distribution of low-frequency alleles suggests a restriction to gene flow outwards from the Mediterranean Sea, across the Straits of Gibraltar.     

Tissue distribution of the amnesic shellfish toxin,domoic acid,in<Emphasis Type="Italic"> Octopus vulgaris</Emphasis> from the Portuguese coast

Domoic acid (DA), the amnesic shellfish toxin, is a food-web-transferred algal toxin that has been detected in many marine organisms from copepods to whales. However, cephalopods, which are important members of the food chain, have never been implicated in DA transfer or accumulation. Here, we present data showing relevant values of DA detected in the common octopus (Octopus vulgaris) from the Portuguese continental coast. Even though DA is hydrophilic and is not expected to be accumulated in the tissues, DA was always detected in our octopus tissue samples. Tissue distribution of DA revealed that the digestive gland and the branchial hearts are the main organs of DA accumulation. Highly variable DA concentrations, ranging from 1.1 to 166.2 g DA g^–1, were observed in the digestive glands. Low levels of DA were detected in the digestive tract (stomach and intestine) and could be a consequence of high digestion rates or a result of non-exposure to toxic vectors during the sampling period. In fact, octopus prey, such as bivalves, crustaceans and fishes, are known to occasionally work as DA vectors. Consequently, DA uptake into octopus tissues is likely sporadic. Similar low levels were detected in the kidney, gills, systemic heart, posterior salivary glands and mantle, and no DA was found in either the gonads or the ink sac. These data are the necessary first step towards achieving an understanding of the accumulation of phycotoxins in O. vulgaris.Communicated by S.A. Poulet, Roscoff     

Concentrations of cadmium,manganese, copper,zinc, and lead in the tissues of the oyster (Crassostrea iredalei) obtained from Setiu Lagoon,Terengganu, Malaysia

Heavy metals pollution in aquatic environments is a major problem contributing to human health issues. The study of these pollutants through bioindicators such as the oyster Crassostrea iredalei is important for (1) determining the levels and sources and (2) regulating the quantity of pollutants. The concentrations of cadmium (Cd), manganese (Mn), copper (Cu), zinc (Zn), and lead (Pb) in tissues of C. iredalei, sediment and surrounding water was measured, and data was analyzed to determine the relationship between sampling periods and between oyster tissue, sediment, and water. The highest concentration of metals in oyster tissue was Zn, followed by Cu, Mn, Cd, and Pb. Concentrations of Cd, Cu, and Zn exceeded the maximum level allowed according to the Malaysian Food Act of 1983, which is equivalent to the WHO recommended levels of heavy metals in organisms used for consumption. The highest metal concentration in sediment was Mn followed by Zn, Pb, Cu, and Cd. Concentrations of heavy metals in surrounding water were Zn, Pb, Cu, Mn, and Cd. There was no correlation between metal concentration in oyster tissue and in sediment for all five metals.     

Preparation and evaluation of protein microcapsules for a marine suspension-feeder,the Pacific oyster Crassostrea gigas

Little is known of the dietary requirements of marine suspension-feeders, mainly due to difficulties in preparing and delivering microparticulate diets to animals living in an aquatic environment. The food particles must be stable in seawater and digestible by the consumer. A laboratory method for preparing cross-linked, protein-walled capsules is described in this study as a means of delivering dietary protein to marine suspension-feeders such as the oyster Crassostrea gigas. Leakage of ¹⁴C-labelled protein from capsules suspended in seawater for 24 h was reduced to less than 5% by the use of the trifunctional cross-linking agent 1,3,5-benzenetricarbonyl trichloride in wall formation. Invitro experiments indicated that capsules were susceptible to digestion by proteases and by style extracts of oysters. Oysters fed encapsulated ¹⁴C-labelled protein absorbed ¹⁴C with an efficiency of 40.2±5.0% (±1 SE). Biochemical fractionation of capsule-fed oysters indicated breakdown of ingested ¹⁴C-protein into material that was soluble in 7% trichloroacetic acid. Cross-linked, protein-walled capsules can be used successfully to deliver dietary protein to the oyster C. gigas.     

Inducible defenses in the eastern oyster Crassostrea virginica Gmelin in response to the presence of the predatory oyster drill Urosalpinx cinerea Say in Long Island Sound

The response of the eastern oyster C. virginica to the presence of the oyster drill Urosalpinx cinerea was examined from July to September 2011. Several aspects of oyster growth were measured, including wet weight, shell weight, and dorsal shell area for oysters collected near Groton, Connecticut (41.32036 N, −72.06330 W). Wet weight and shell weight growth were significantly higher in the presence of the predator U. cinerea, while tissue weight showed no difference from the control. The control group showed more shell area growth and a much lower ratio of shell weight growth to shell area growth. Differences in shell weight to area ratio indicated that C. virginica dramatically shifted from lateral shell growth to shell thickening in the presence of U. cinerea. This inducible defense has not been previously shown for C. virginica and could play an important role in the predator–prey interaction between these two species.     

Sequence divergence of mitochondrial DNA indicates cryptic biodiversity in Octopus vulgaris and supports the taxonomic distinctiveness of Octopus mimus (Cephalopoda: Octopodidae)

DNA sequence diversity of octopods was investigated using the mitochondrial cytochrome oxidase III gene (mtCOIII). DNA was obtained from ethanol- or formalin-fixed tissue of 15 specimens belonging to Octopus mimus Gould, 1852, Octopus vulgaris Cuvier, 1797 and Scaeurgus unicirrhus d'Orbigny, 1840, from coastal waters of the Mediterranean (France), the southwestern Atlantic (Brazil), the Caribbean (Costa Rica) and the southeastern/tropical Pacific (north Chile/Costa Rica). A 612 bp fragment of the mtCOIII gene was sequenced and aligned to the orthologous sequences available from northeastern Pacific Octopus species. Possible phylogenetic relationships were reconstructed. The mtCOIII gene tree revealed two main clusters, one comprised O. rubescens, O. dofleini and O. californicus, while the other included all O. vulgaris specimens, O. bimaculatus, O. bimaculoides and O. mimus. With one exception all South American haplotypes including O. bimaculatus and O. bimaculoides appeared as the sister group of the Mediterranean haplotype of O. vulgaris, indicating that most of the South American O. vulgaris specimens investigated would not belong to the recently redescribed species O. vulgaris restricted to the Mediterranean and eastern Atlantic. The treatment of O. mimus as a species distinct from Mediterranean O. vulgaris is supported by a high nucleotide divergence of 12.7%. Based on the mtCOIII gene tree the existence of cryptic species among O. vulgaris-like octopods is suggested. Received: 15 January 1999 / Accepted: 15 October 1999     

Stable isotopes (δ13C, δ15N) and modelling as tools to estimate the trophic ecology of cultivated oysters in two contrasting environments

Food sources for cultivated marine bivalves generally are not well identified, although they are essential for a better understanding of coastal ecosystems and for the sustainability of shellfish farming activities. In addition to phytoplankton, other organic matter sources (OMS), such as microphytobenthos and detritus (of terrestrial or marine origins), can contribute significantly to the growth of marine bivalves. The aim of this study was to identify the potential food sources and to estimate their contributions to the growth of the Pacific oyster (Crassostrea gigas) in two contrasting trophic environments of Normandy (France): the Baie des Veys (BDV) and the Lingreville area (LIN). Two sites were studied in the BDV area (BDV-S and BDV-N) and one in the LIN area. To estimate the contribution of each type of OMS, we used a combination of stable natural isotope composition (δ¹³C, δ¹⁵N) analysis of OMS and oyster tissue together with a modelling exercise. Field sampling was conducted every 2 months over 1 year. The sampled sources were suspended particulate organic matter from marine (PhyOM) and terrestrial (TOM) origins, microphytobenthos (MPB), detrital organic matter from the superficial sediment (SOM), and macroalgae (Ulva sp., ULV). A statistical mixing model coupled to a bioenergetic model was used to calculate the contributions of each different source at different seasons. Results showed that isotopic composition of the animal flesh varied with respect to the potential OMS over the year within each ecosystem. Significant differences were also observed among the three locations. For instance, the δ¹³C and δ¹⁵N values of the oysters ranged from −20.0 to −19.1‰ and from 6.9 to 10.8‰ at BDV-S, from −19.4 to −18.1‰ and from 6.4 to 10.0‰ at BDV-N, and from −21.8 to −19.4‰ and from 6.3 to 8.3‰ at LIN. The contributions of the different sources to oyster growth differed depending on the ecosystem and on the period of the year. Phytoplankton (PhyOM) predominated as the principal food source for oysters (particularly in the LIN location). MPB, TOM, and ULV detritus also possibly contributed to oysters’ diet during summer and autumn at the BDV-S and BDV-N sites. SOM was not considered an OMS because it was already a mix of the other four OMS, but rather a trophic reservoir that potentially mirrored the trophic functioning of marine ecosystems.     

A whole-animal bioassay for the determination of the food attractants of the lobster Homarus gammarus

A semi-quantitative assay is described for assessing the effectiveness of bait extracts for the lobster Homarus gammarus (L.), using an extract of the squid Loligo vulgaris Lamarck as a standard. Fractionation of this extract indicated that, while no single component was as attractive to lobsters as the whole, the recombination of the various components yielded a solution which was as attractive as the original extract. It appears likely that several different classes of chemosensory cell must be simultaneously stimulated for the initiation of feeding behaviour in the lobster.     

Morphological and habitat divergence in the intertidal limpet Patelloida pygmaea

In the intertidal limpet Patelloida pygmaea, two distinctive morphs, the forms pygmaea and conulus, have been recorded. The former possesses a flat elongated shell, and the latter has an extremely high round shell. It has been observed in the field that pygmaea is found on oyster shells Crassostrea gigas. The form conulus uses an unusual substrate for attachment. It is found on the living shells of the intertidal gastropod Batillaria cumingi. Although conulus is normally found only on shells of Batillaria, it can also be found on oyster shells when pygmaea and Batillaria shells are not present in nature. An electrophoretic analysis of allozymes showed that these two forms are reproductively isolated from each other and coexist without gene exchange on the same mudflat. Laboratory experiments showed that pygmaea prefers oyster shells and conulus prefers Batillaria shells as substrates for attachment when both oyster and Batillaria shells are present. The form pygmaea did not attach to Batillaria shells, even when only Batillaria shells were available. However, conulus also attached to oyster shells when Batillaria shells were not available. The proportion of individuals of conulus that attached to oyster shells decreased significantly when pygmaea was attaching to the oyster shells. These results suggest that pygmaea is ecologically more specialized to living on oyster shells than conulus.     

Alongshore transport of a toxic phytoplankton bloom in a buoyancy current: Alexandrium tamarense in the Gulf of Maine

We examined the mechanisms controlling blooms of the toxic dinoflagellate Alexandrium tamarense Lebour and the concomintant patterns of shellfish toxicity in the southwestern Gulf of Maine, USA. During a series of cruises from 1987 to 1989, hydrographic parameters were measured to elucidate the physical factors affecting the distribution and abundance of dinoflagellates along this coast. In 1988 and 1989 when toxicity was detected in the southern part of this region, A. tamarense cells were apparently transported into the area between Portsmouth, New Hampshire, and Cape Ann, Massachusetts, in a coastally trapped buoyant plume. This plume appears to have been formed by the outflow from the Androscoggin and Kennebec Rivers. Flow rates of these rivers, hydrographic sections, and satellite images led us to conclude that the plume persisted for about a month, and extended alongshore for several hundred kilometers. The distribution of cells followed the position of the plume as it was influenced by wind and topography. When winds were downwelling-favorable (to the southwest), cells were moved alongshore to the south, and were held to the coast; when winds were upwelling-favorable (to the northeast),the plume sometimes separated from the coast, advecting the cells offshore. In 1987 when no plume was present, A. tamarense cells were scarce, and no toxicity was recorded at the southern stations. The alongshore advection of toxic cells within a coastally trapped buoyant plume can explain the details of the temporal and spatial patterns of shellfish toxicity along the coast. We hypothesize that (1) the source of the A. tamarense populations is in the north, possibly associated with the Androscoggin and Kennebec estuaries, that (2) toxicity patterns follow a predictable relationship with river flow volume and timing of flow peaks and that (3) wind stresses directly influence the distribution of low salinity water and the dinoflagellate cells. Local, in situ growth of dinoflagellates can be an important factor initiating toxic dinoflagellate blooms. However, these data demonstrate the significant role of alongshore transport of established populations of A. tamarense in controlling the location and timing of paralytic shellfish poisoning (PSP) outbreaks in May and June along the southwestern coast of the Gulf of Maine.     

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.     

Exploring the role of chitinolytic enzymes in the sea fan coral, <Emphasis Type="Italic">Gorgonia ventalina</Emphasis>

Chitinases are involved in defense against chitinaceous pathogens in both invertebrates and vertebrates. This study investigated whether sea fan corals, Gorgonia ventalina (Linnaeus) collected from the Florida Keys between the summer of 2002 and the summer of 2005 contain chitinases, and whether these enzymes could serve an analogous protective role against the fungal pathogen, Aspergillus sydowii (Thom et Church). Crude extracts of healthy sea fans contained detectible levels of exochitinase activity in an in vitro microplate assay using fluorogenic substrates. The exochitinase levels decreased upon injury, agitation, or manipulation of the tissue. A concurrent, transient increase of exochitinase in the surrounding water suggests that sea fans release chitinases as a response to these stresses. By contrast, endochitinase was detected in only 2 of 15 sea fans (13%), suggesting a high degree of variation for this enzyme. Sea fan chitinase-containing seawater and anion exchange chromatography fractions were both active against A. sydowii in an absorbance-based antifungal assay. The presence of chitinases in sea fan extracts, their release into the surrounding water upon stress, and their activity against A. sydowii suggests that further study of these enzymes in coral stress responses is warranted.     

Avoiding predation: alarm responses of Caribbean sea urchins to simulated predation on conspecific and heterospecific sea urchins

Alarm responses to the extracts of conspecifics and hetero-specifics were measured for the Caribbean sea urchins Echinometra viridis, E. lucunter, Lytechinus variegatus, L. williamsi, Tripneustes ventricosus, Diadema antillarum, and Eucidaris tribuloides collected along the Caribbean coast (9°3314N; 78°5523W) during October 1984 and July–December 1985. Responses to seawater and extracts of the gnathostomate echinoid Clypeaster sybdepressus were used as controls. The intensity of the response resulting from exposure to sea-urchin extracts was measured by: (1) the percentage of individuals that responded by moving away from the extract and/or towards shelter, and (2) the mean distance moved. Echinometra viridis, E. lucunter, and L. williamsi responded to sea-urchin extracts by moving towards nearby shelter sites. The distance that individuals of each species moved in the first minute following exposure to conspecific extracts was correlated with the distance that species moved from shelter while foraging. L. variegatus and D. antillarum, living in microhabitats not providing protection from predators, responded to extracts of conspecifics and heterospecifics by moving away from the direction of the extract. Eucidaris tribuloides did not exhibit alarm responses to the extracts of con- or hetero-specifics. E. tribuloides secures itself with its stout spines into protected sites within corals. Similarly, L. variegatus living in long, dense seagrass that provided protection from detection by predators, and D. antillarum occupying crevices, showed no alarm responses to extracts of conspecifics. Presumably, in these situations, sea urchins cannot increase their defenses against predation by moving away from an injured neighbor. T. ventricosus showed a weak response to extracts of L. variegatus, but no response to extracts of other species including conspecifics. The reasons for this lack of an alarm response are unclear. For the five species that demonstrated an alarm response to sea-urchin extracts, the intensity of the response varied depending on the type of extract used. L. variegatus, L. williamsi, and D. antillarum responded most strongly to extracts from conspecifics, while Echinometra viridis and E. lucunter responded strongly to extracts from both conspecifics and congeners. The weakest responses were shown to the extracts of T. ventricosus and Eucidaris tribuloides. Habitat overlap, overlap in predators, and phylogenetic relationships did not consistently explain patterns of alarm responses to the extracts of heterospecific sea urchins.     

Karyotype, nucleolus organiser regions and constitutive heterochromatin in Ostrea angasi (Molluscae: Bivalvia): evidence of taxonomic relationships within the Ostreidae

Chromosome preparations from gill tissue of the Australian flat oyster Ostrea angasi Sowerby were studied with conventional Giemsa staining, C-banding and silver staining techniques. A diploid complement of 2n = 20 was observed, consisting of five metacentric, three submetacentric and two subtelocentric pairs. Constitutive heterochromatin was distributed as large centromeric blocks in Chromosome Pairs 3, 6, 8, 9 and 10. The nucleolus organizer regions (NORs) were located terminally on long arms of Chromosome Pairs 9 and 10. This allowed the identification of homologous chromosomes in submetacentric and subtelocentric pairs. Intraspecific variability in NOR pattern as revealed by differences in the number of silver-stained NORs (Ag-NORs) per cell was found to be very common. Comparison of the patterns of karyotype, C-band and Ag-NORs between species of the larviparous oysters for which data have been published demonstrate that the chromosomal structure of the endemic Australian and New Zealand species O. angasi shows little similarity to the Southern Hemisphere oysters O. (Eostrea)puelchana Orbigny and Tiostrea chilensis (Philippi) and the Indo-West Pacific oyster O. denselamellosa, but very high resemblance to the European species O. edulis. Received: 12 August 1996 / Accepted: 16 September 1996