The common mussel Mytilus edulis as an indicator of trace metals in Scandinavian waters. II. lead,iron and manganese

An investigation of the mechanisms of detoxication of copper and zinc by the oyster Ostrea edulis (L) has been carried out using naturally occurring green-sick (contaminated by copper) and unpolluted oysters. Electron microprobe X-ray analysis of tissues in the electron microscope gives direct evidence for the structural compartmentation of copper and zinc in separate, specific, granular amoebocytes. The metals are immobilized in membrane-limited vesicles as different chemical compounds, copper being associated with sulphur and zinc with phosphorus. Chemical analyses of serum and tissues of normal and green-sick oysters indicate that (a) Cu and Zn are accumulated independently, (b) the Cu and Zn in the serum, while higher than in the surrounding sea water, are maintained at a 10-fold smaller level than the tissues, (c) toxicity is reduced by active uptake from the serum into granular amoebocytes, where it is further reduced by compartmentation in membrane-limited vesicles. It is calculated that the individual cell types may contain as much as 13,000 ppm Cu and 25,000 ppm Zn.     

Opsonin-like molecule found in coelomic fluid of a sea cucumber, Holothuria leucospilota

Opsonins, a group of serum proteins present in vertebrates, are known to bind to the surface of many kinds of pathogens and tag them as targets for phagocytosis. Opsonin-like molecules have been suggested to exist in several invertebrates. In this paper, we report the discovery of a new opsonin-like molecule in the coelomic fluid of the sea cucumber Holothuria leucospilota. Opsonic activity in cell-free coelomic fluid was characterized by sodium dodecyl sulfate–polyacrylamide gel electrophoresis, two-dimensional electrophoresis and gel filtration. Phagocytic uptake of yeast cells and latex beads by amoebocytes was quantified by microscopic analysis. It was found that a 20 kdalton protein bound to and enhanced phagocytosis of heat-killed yeast cells. The results of Western blot analysis suggest that both amoebocytes and spherule cells produce this opsonin-like molecule. Received: 22 October 1998 / Accepted: 4 March 2000     

Photosynthetic and optical properties of the marine chlorophyte Dunaliella tertiolecta grown under fluctuating light caused by surface-wave focusing

Photosynthetic and optical properties of the marine chlorophyte Dunaliella tertiolecta Butcher were studied in response to irradiance fluctuations caused by surface-wave focusing. The experimental conditions simulated the prominent features of the light field (high average irradiance, spectral composition and statistical properties) in the uppermost few meters of the water column under sunny surface conditions. The properties of algae grown under high-frequency fluctuations were compared with control cells grown under constant light at the same average irradiance (800 mol quantam^-2s^-1). No significant differences were found for a number of parameters, including growth rate, cellular chlorophyll a and pigment ratios, photosynthetic unit size and density of Photosystem I reaction centers, the rate of photosynthesis at the growth irradiance, dark respiration, and in vivo fluorescence of chlorophyll a per cell. Photosynthetic parameters were not affected by whether the incident light for oxygen exchange measurements was fluctuating or constant. This was the case whether the cells had been previously acclimated to either fluctuating or constant irradiance. Such a photosynthetic response indicates that cells are accomplishing a time integration of the fluctuating light. In addition, although D. tertiolecta is capable of dramatically changing its optical properties in response to low or high growth irradiance levels, the refractive index of the cells, the efficiency factors for light absorption and scattering by individual cells, and chlorophyll-specific absorption and scattering coefficients of cell suspensions, were all very similar under high irradiance, whether or not wave focusing was present.Contribution to the program of GIROQ (Groupe Interuniversitaire de Recherches Océanographiques du Québec)     

Patterns of growth and triacylglycerol content in snow crab Chionoecetes opilio (Brachyura: Majidae) zoeal stages reared in the laboratory

Snow carb Chionoecetes opilio zoea I and zoea II larvae, hatched from females in a controlled mating experiment, were reared in the laboratory at 10.1 °C and 28.0 salinity, to resolve the patterns of growth (dry weight [DW]) and change in energy reserves (triacylglycerols [TAG]) within a given moult cycle. The patterns of growth and change in TAG reserves were similar in each zoeal stage. Following hatching or a moult, the zoeae entered a phase of rapid size increase, i.e. high daily growth rates (5.5 to 12.8% DWd^-1), for 1/3 to 2/5 of the duration of the moult cycle. During the same period, the zoeae accumulated TAG reserves until a maximum (TAG DW^-1) was reached at the end of the phase of rapid growth. The period of high growth rates and of TAG accumulation is interpreted as the required time for the zoeae to reach a point in development [i.e. point of reserves saturation (PRS); Anger and Dawirs (1981)] where sufficient growth and energy reserves allow moulting to the next stage. Following the phase of rapid growth and TAG accumulation, the zoeae entered a phase of low daily growth rates (0 to 1% DWd^-1) during which the TAG reserves decreased to a minimum at the end of the phase. Prior to, and during the moult to zoea II, a phase of negative growth was observed in the zoea I larvae. We conclude that measurement of zoeal size and TAG content, along with morphometric criteria (e.g. epidermal retraction), can be used to assess growth and nutritional condition of C. opilio zoeal stages from the sea.     

A light and electron microscope investigation of gametogenesis in Nicolea zostericola (Polychaeta: Terebellidae)

A light and electron microscope study was conducted on Nicolea zostericola (Oersted: Grube, 1860) to describe the processes of gametogenesis. N. zostericola is a monotelic species which spawns from 2 to 5 times over a period of 2 weeks at the end of its 1-year life span. The gonads consist of plates of cells bulging from beneath the peritoneum in the thoracic region of the body. Oogenesis has been divided into 3 phases: an ovarian phase in which gametogonia and primary oocytes are retained within the ovary, a follicular phase, in which oocytes are ovulated into the coelom surrounded by several layers of follicle cells and the vitellogenic phase, where individual oocytes rupture through the follicle cells and enter the coelomic fluid where cell growth and yolk synthesis occur. The combined ovarian and follicular phases have a duration of about 3 to 5 weeks and the vitellogenic phase about 5 to 6 months. Vitellogenesis is described on the ultrastructural level. Yolk synthesis involves the participation of the Golgi apparatus and endoplasmic reticulum of the oocyte in possible combination with nutrient absorption from the coelomic fluid. Spermatogenesis involves the rupturing of masses of spermatogonia into the coelom from the testis, surrounded by peritoneal cells. Smaller clusters of spermatocytes are later released into the coelomic fluid where mitotic and meiotic divisions eventually produce flattened plates of sperm. The mature spermatozoan possesses an elongate head, atypical for most polychaetes.     

Modification of the feeding behavior of marine copepods by sub-lethal concentrations of water-accommodated fuel oil

The feeding behaviors of Acartia clausi and A. tonsa were measured in samples of water containing low levels of a water-accommodated fraction of No. 2 fuel oil. The copepods fed normally at a hydrocarbon concentration of 70 g l^-1, but their feeding behavior was altered both quantitatively and qualitatively at a concentration of 250 g l^-1. Three types of response to the higher oil level were found. The first was total suppression of feeding. Both other types involved suppression of feeding on particles between 7 and 15 m diameter, but one showed no change in the ingestion of larger particles, whereas the other displayed increased feeding on particles larger than 15 m diameter. These results suggest that the species of Acartia studied use three different modes of feeding, each on a different size range of particulate material. Low-level hydrocarbon pollution affects each feeding mode differently.Contribution No. 973, Center for Environmental and Estuarine Studies of the University of Maryland     

Oxygen dependent sulfide detoxification in the lugwormArenicola marina

The lugwormArenicola marina L. oxidizes entering sulfide to thiosulfate. After 8 h of normoxic incubations with sulfide concentrations of 0.2 to 1.0 mmoll^-1 thiosulfate in the coelomic fluid amounted up to about 4 mmoll^-1 whereas sulfite concentrations were 100-fold lower and no accumulation of sulfate in the coelomic fluid was found. The sulfide oxidation was highly oxygen dependent. An increase of oxygen partial pressure ( ) in the medium was followed by enhanced thiosulfate production and by a decrease of sulfide concentration in the coelomic fluid. Under normoxia, the sulfide oxidation rate was sufficient to compensate the influx of sulfide into the coelomic fluid when the sulfide concentration in the medium was below 0.33 mmoll^-1. When external sulfide was raised beyond this level, sulfide up to 5 moll^-1 in the coelomic fluid appeared. Succinate in the body wall tissue was low as long as no sulfide appeared in the coelomic fluid, indicating the maintenance of an aerobic metabolism. The oxidation of sulfide to thiosulfate was localized in the mitochondria of the body wall tissue. The oxygen consumption of mitochondria was stimulated by the addition of sulfide. The mitochondrial sulfide oxidation rate depended on the amount of mitochondrial protein and followed a Michaelis-Menten kinetic. An apparentK _m of 0.68±0.29 moll^-1 and aV _max of 41.9±22.3 nmol min^-1 mg^-1 protein was calculated. Sulfide was stoichiometrically oxidized to thiosulfate with 1 mol sulfide consuming 1 mol oxygen. Sulfide oxidation was not inhibited by sulfide concentrations as high as 100 moll^-1. At low concentrations of cyanide or azide, when respiration without sulfide was already inhibited, sulfide oxidation could still be stimulated, tentatively indicating the existence of an alternative terminal oxidase. Specimens examined in the present study were collected near St. Pol de Leon, France, from 1989 to 1992.     

Flocking behaviour of passerines: a dynamic model for the non-reproductive season

Summary The behaviour of a small male passerine bird over a typical winter day is studied by a dynamic programming model. The bird can be either unpaired or paired; an unpaired bird can forage in a flock, forage alone or sing to attract a mate. Foraging increases his reserves, while singing reduces them. The optimal policy and the expected behaviour of birds depend both on time and reserves. The model predicts that birds will flock, especially in the morning, if flocking birds find more food (foraging efficiency), and also more flocking can be expected when the predation risk is lower in a flock (antipredator benefit). Where flocking gives lower variance in food intake, with the same mean (reduced variance benefit), birds with low reserves at the end of the day choose to forage alone (high variance option), while otherwise they are risk-averse and forage in a flock. The cost of flocking increases with time in a day and with the probability of mate attraction through singing. Decisions inevitably involve trade-offs. Where flocking results in antipredator benefit, but also lower foraging gain, birds with low reserves forage alone, but birds with high reserves flock. Offprint requests to: A.I. Houston     

Ultrastructure of the gill of the hydrothermal-vent mytilid Bathymodiolus sp.

Specimens of Bathymodiolus sp. were collected at 2 620 m depth during the Biocyarise Cruise (12°5880N; 13°5660W) in March 1984, and samples of the gill were fixed for ultrastructural observations. The gill of this hydrothermal-vent mytilid constitutes the main organ in the nutritional processes. The lamellae display abundant ciliation, normally comprised of frontal cilia, compound latero-frontal cirri and lateral cilia. At the ventral margin of each demi-branch, a longitudinal, ciliated, feeding groove is present. the lamellae are composed of numerous homorhabdic filaments connected by tufts of cilia. Each filament is made of a thin wall overlying a central lumen containing amoebocytes. Ultrastructural observations revealed the filament wall to be composed of four types of cells: (1) The ciliated cells of the frontal, latero-frontal and lateral ciliation, characterized by an abundance of mitochondria. (2) Mucous cells present to some degree among the ciliated cells, but more abundant on the distal edge and containing dense droplets of mucus. (3) Cells colonized at their apical pole by numerous bacteria enclosed in membrane-delimited clear spaces and composing the major part of the filament wall. (4) Thin ciliated cells separating the bacterial cells and characterised by a dense fringe of microvilli at their apical pole. The lumen of the filament contains amoebocytes of different morphological aspects which seem to accumulate electron-dense granules, possibly related to detoxification processes.     

Metabolic and blood characteristics of the hydrothermal vent tube-worm Riftia pachyptila

Specimens of the hydrothermal vent pogonophoran Riftia pachyptila Jones were collected by submersible at a depth of 2 600 m at the 21°N hydrothermal vent site on the East Pacific Rise (20°50N, 109°06W) in April and May of 1982. The worms were maintained in pressurized aquaria for up to 45 d for metabolic studies. Consumption of O₂ was regulated down to low P_{O 2} (oxygen partial pressure) values; O₂ consumption rates were 0.63 and 1.12 mol g^-1 wet wt h^-1 at 2.5° and 8°C, respectively; such rates were comparable to those previously measured for other pogonophorans. Intact specimens of R. pachyptila (including bacterial symbionts) did not consume significant amounts of CH₄ from the environment. The respiratory quotients, in the absence of added sulfide, indicated that metabolism was mainly heterotrophic. High rates of uptake of dissolved amino acids were recorded for one specimen. The total [CO₂] in the vascular blood and the Hb-containing coelomic fluid were high. Under anaerobic conditions, there were equilibrium distributions of pH, total [CO₂] and sulfide concentrations between the vascular blood and the coelomic fluid, apparently because these metabolites were readily exchanged between the two compartments. The vascular blood bound neither CH₄ nor H₂. However, sulfide was reversibly bound by both the vascular blood and coelomic fluid; because this binding depended strongly on pH (with a maximum at about 7.5), HS^- was probably the molecular species bound. Under anaerobic, but not aerobic conditions, the trophosome bound substantial amount of sulfide; thus, the high concentrations of sulfide in the trophosome may have resulted mainly from sulfide bound to sulfide oxidases under anaerobic conditions. The coelomic fluid had a relatively low buffering capacity (2.2 mmol CO₂pH^-1).     

Reproductive cycle of Golfingia margaritacea,a bipolar sipunculan,in subantarctic water

The reproductive cycle of the bipolar sipunculan Golfingia margaritacea collected from Ushuaia, Argentina (54°49 S; 68°13W) in 1986 and 1987 is reported. The population sampled showed a sex ratio of 1:1. Males, however, frequently outnumbered females. Measurements of coelomic oocytes delineated an annual reproductive cycle. Small oocytes were present in the coelom all year: a subpopulation began to increase size about the middle of spring and reached its maximum size in late summer. Spawns may occur from the end February to April. May is characterized by resorption of large oocytes. Monthy monitoring of the gonads revealed mitotic activity and a continuous release of gametocytes to the coelom. Parasites were detected in the gonads. Large coelomic oocytes were also present in the coelom during the same season in the Northern and Southern Hemispheres, but egg sizes were significantly larger in boreal populations.     

Experimental test of an allometric method for estimating potential copepod production

Phytoplankton and zooplankton, collected in November 1984 at the deepest point of Bedford Basin, Canada, were used in a 2 mo experiment in the Dalhousie tower tank to test the validity of estimating potential copepod production from allometric equations based on body size and temperature under food-saturated conditions (maximum instantaneous growth, MIG method). The development of the plankton community in the mesocosm was similar to fall and winter assemblages in a coastal Nova Scotian inlet. Predicted instantaneous rates of potential production were compared with average production of a mixed copepod assemblage measured over weekly intervals in the absence of predators. Predictions exceeded measured production by 1.9 to 136 times, but generally agreed within an order of magnitude. Whereas the MIG method facilitates automated estimation of potential copepod production, this advantage must be offset by the relative loss of accuracy compared to more labour-intensive methods. The method appears to be most useful for mesoscale surveys or neritic regions where the aim is to quickly obtain an order of magnitude estimate of secondary production.     

Night irradiance and synchronization of lunar release of planula larvae in the reef coral Pocillopora damicornis

Pocillopora damicornis (Linnaeus), which is known to release planula larvae on a monthly cycle, was grown in full daytime solar irradiance, but with four treatments of night irradiance: (1) natural night irradiance, (2) shifted-phase (total darkness during nights of full moon with artificial irradiance at lunar intensity on nights of new moon), (3) constant full moon (full lunar irradiance every night), and (4) constant new moon (total darkness every night). The reproductive cycle of the corals held in the shifted-phase treatment moved out of synchrony with the cycle of corals exposed to a natural lunar cycle of night irradiance. Two previously described types of P. damicornis were tested. The Type Y normally start releasing larvae at full moon, with peak production at third quarter. In the shifted-phase treatment they began releasing planulae at new moon (artificial full moon), with peak production at first quarter. The Type B corals, that normally start releasing planulae at new moon with peak production at first quarter, began to release planulae at full moon (artificial new moon), with peak production at third quarter. Populations of corals grown either in the constant full moon or constant new moon treatment quickly lost synchronization of monthly larva production, although production of planulae continued. Thus spawning is synchronized by night irradiance.Contribution No. 702 of the Hawaii Institute of Marine Biology     

Blue-winged warblers (Vermivora pinus) “recognize” dialects in type II but not type I songs

Summary Male blue-winged warblers (Vermivora pinus) sing two distinctly different song types (I and II). Type II songs from eastern New York and western Massachusetts, USA, are different (Fig. 1), and during playback experiments males respond stronger to local than to foreign songs (Fig. 2). Type I songs are more stereotyped geographically (Fig. 1), and males do not differentiate between local and foreign Type I songs (Fig. 3). Differing degrees of intra- and inter-sexual selection on these two forms may have promoted variability in Type II songs and stereotypy in Type I songs among locations.     

Studies on the fusion reaction in two species of Perophora (Ascidiacea)

Histological and electron microscopical studies were made on the fusion reaction between homogeneic stolons of the colonial ascidians Perophora japonica Oka and P. sagamiensis Tokioka. When two stolons make a tip-to-side contact, the epidermal cells of the lateral portion of one stolon become thick and form a protrusion. In the fusion reaction of P. japonica, epidermal cells of each stolon at the contact area extend cell processes immediately after contact. After an occurrence of degenerated epidermal cells, blood interchange between the two stolons is initiated by a partial elimination of the epidermal cells. Even after the establishment of blood interchange, the cuticlelike structure is often observed to remain at the contact area. In the fusion reaction of P. japonica, many amoebocytes are attached to the epidermal cells until the complete elimination of epidermal cells. In the fusion reaction of P. sagamiensis, many lymphocytes as well as amoebocytes are attached to the epidermal cells at the contact area.Contribution No. 461 of the Shimoda Marine Research Center     

Temporal dynamics and plasticity in the cellular immune response of the sea fan coral, Gorgonia ventalina

The temporal dynamics of the invertebrate immune response often determines an organism’s success in responding to physiological stress, physical damage, and pathogens. To date, most immune challenge studies have been conducted under highly controlled laboratory conditions, with few attempts to study immune function in the wild. In this study, we characterized the temporal dynamics of the Caribbean sea fan, Gorgonia ventalina, cellular immune response [granular amoebocyte aggregation and prophenoloxidase (PPO) activation] to allogenic grafts in the laboratory and field using a clonally replicated design. Amoebocyte reaction time differed markedly between the lab (% amoebocyte surface area in tissue sections peaked at 2 days) and field (peak at 6 days). PPO activity decreased between 0 and 6 days after grafting in both ungrafted and grafted tissue, suggesting PPO is decoupled from other cellular components. The reaction norms of the fold induction in % amoebocyte area between disease-grafted and healthy-grafted tissue of each colony across time indicate high intercolony plasticity in cellular immune response. The plasticity between colonies was also evident in the magnitude of cellular immune response, ranging from a 0.88- to 1.60-fold increase in amoebocyte area between initial and 6 days for the disease-grafted tissue. With the demonstration of highly dynamic cnidarian cellular immune responses, our study expands understanding of the evolutionary ecology of metazoan immune defense mechanisms.     

Examples of post-mortality alteration in Recent brachiopod shells and (paleo)ecological consequences

Post-mortality alteration of brachiopod shells under normal environmental conditions leads to high taphonomic loss, and to a poor contribution to the biodetrital soft sediment. The successive stages of alteration which shells undergo are: (bio)degradation of the organic matrix shell softening structural disaggregation or/and mechanical fragmentation; these processes depend on the shell structure (number of layers) and composition (organic and inorganic components), but very little on environmental conditions, except for the dissolution of inorganic shell constituents. Among the Brachiopoda, three types of alteration occur to different types of shells — Type I: two-layered chitino-phosphatic shell (species ofLingula andGlottidia) displays a rapid degradation of the organic matrix and mechanical abrasion, leading to total disappearance of the shell in 2 to 3 wk; Type II: two-layered carbonate shell, e.g.Terebratulina spp. [but notNotosaria nigricans (Sowerby) which may constitute a fourth shell type], exhibits degradation of the organic matrix of the secondary layer, shell softening, and structural disaggregation leading to shell disintegration in 6 to 7 mo with a concomitant contribution of calcitic microfibres to the sediment; Type III: three-layered carbonate shell, e.g.Gryphus vitreus (Born), undergoes organic degradation of the secondary layer, fragmentation of the anterior two-thirds of the shell, and slow degradation (because of the thick tertiary layer) of the posterior portion of the shell, with dissolution of the inorganic components (mainly in the tertiary layer) which make a relatively minor contribution to the sediment.     

Nucleic acid and protein levels in strobilating polyps of Chrysaora quinquecirrha and Aurelia aurita

Levels of DNA, RNA, and protein were measured in scyphistomae of the scyphozoans Aurelia aurita and Chrysaora quinquecirrha during strobilation. In synchronously developing populations of A. aurita, the amount of DNA per polyp increases 41 to 260% during strobilation, whereas RNA and protein remain relatively constant. The RNA/DNA ratio drops by about 50% during strobilation in both C. quinquecirrha and A. aurita, and protein/DNA decreases by 50% in A. aurita. Specific RNA reserves are probably not accumulated in advance of strobilation, since strobilation is blocked immediately in both species by 1 to 3 g/ml actinomycin D. The size and DNA content of polyps are usually increased by prolonged incubation at temperatures of 12° to 15°C, which facilitates strobilation; however, the RNA/DNA and protein/DNA ratios of such polyps are not significantly increased by the cold conditioning.Contribution No. 472 from the Virginia Institute of Marine Science.     

An electron-microscope study of periostracum repair in Mytilus edulis

The ultrastructural details of changes that occur in the inner face of the outer fold of Mytilus edulis during periostracum repair are described. Initially, after the periostracum is slit, increased secretory activity occurs in the cells, apparently controlled by lysosomes. Proto-ostracal material deposited outside the cells seems to be due to Golgi secretory granules, which dissociate and then re-assemble outside the cells to form an organised layer. During the later stages of repair, autophagic vacuole formation increases in the cells, possibly because of increased demands for synthetic material. When substantial amounts of proto-ostracal material are secreted and the outer fold is sealed off from the external environment, the cells return to their normal condition. During the early stages of repair, amoebocytes appear to be implicated in the laying down of proto-ostracal material.