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.     

Elemental composition of the hydrothermal vent clam Calyptogena magnifica from the East Pacific Rise

Specimens of the hydrothermal vent clam Calyptogena magnifica were collected at a depth of 2 600 m from 21° N on the East Pacific Rise in spring, 1982. The elemental composition was determined for the total soft tissues, individual organs, and the shell. The soft parts contained high concentrations of a number of trace metals. Iron, copper, and zinc, at 760, 148 and 2 152 g g^-1 dry weight, respectively, were present at the highest concentrations. Silver and antimony exhibited the greatest enrichments when their concentrations in C. magnifica were compared with those measured in a shallow-water marine mussel, Mytilus edulis. In contrast to the soft parts, the shell was not a site of metal accumulation or deposition. Within the soft parts; gills, kidney, pericardium, and mantle were involved in the concentration of various metals; the degree of concentration varied depending on the particular metal under consideration. C. magnifica possesses metal body burdens which are ordinarily considered to be very high and potentially toxic in other species.     

Sexual reproduction of the solitary sunset cup coral<Emphasis Type="Italic"> Leptopsammia pruvoti</Emphasis> (Scleractinia: Dendrophylliidae) in the Mediterranean. 1. Morphological aspects of gametogenesis and ontogenesis

Information on the reproduction in scleractinian solitary corals and in those living in temperate zones is notably scant. Leptopsammia pruvoti is a solitary coral living in the Mediterranean Sea and along Atlantic coasts from Portugal to southern England. This coral lives in shaded habitats, from the surface to 70 m in depth, reaching population densities of >17,000 individuals m^–2. In this paper, we discuss the morphological aspects of sexual reproduction in this species. In a separate paper, we report the quantitative data on the annual reproductive cycle and make an interspecific comparison of reproductive traits among Dendrophylliidae aimed at defining different reproductive strategies. The present study on L. pruvoti is the first in-depth investigation of the reproductive biology of a species of this genus. As expected for a member of the family Dendrophylliidae, L. pruvoti is a gonochoric and brooding coral. The gastrodermal tissue of the gametogenetic mesenteries we examined was swollen and granular, which led us to hypothesize that interstitial cells could have a trophic function favoring gametogenesis. Undifferentiated germ cells arose in the gastrodermis and subsequently migrated to the mesoglea, where they completed gametogenesis. During spermary development, spermary diameter increased from a minimum of 14 m during the immature stages to a maximum of 410 m during the mature stages. As oogenesis progressed, we observed a gradual reduction in the nucleus to cytoplasm ratio due to the steady synthesis of yolk. During the final stages of oogenesis, after having migrated to the extreme periphery of the oocyte and having firmly adhered to the oolemma, the nucleus became indented, assuming a sickle or dome shape. We can hypothesize that the nucleus migration and change of shape may have to do with facilitating fertilization and determining the future embryonic axis. During oogenesis, oocyte diameter increased from a minimum of 20 m during the immature stage to a maximum of 680 m when mature. Embryogenesis took place in the coelenteron. We did not see any evidence that even hinted at the formation of a blastocoel; embryonic development proceeded via stereoblastulae with superficial cleavage. Gastrulation took place by delamination. Early and late embryos had diameters of 204–724 m and 290–736 m, respectively. When released, the larvae had completed ontogenesis and swam by a ciliary movement with the aboral pole at the anterior, their shape varied from spherical to cylindrical (in the latter the oral–aboral axis measured 695–1,595 m and the transversal one measured 267–633 m).Communicated by R. Cattaneo-Vietti, Genova     

Changes of plasma osmolality,chloride concentration and gill Na−K-ATPase activity in tilapia Oreochromis mossambicus during seawater acclimation

Changes of plasma osmolality, chloride concentration and gill Na–K-ATPase activity in tilapia Oreochromis mossambicus (obtained from Tainan Fish Culture Station of Taiwan Fisheries Research Institute, 1987) during seawater acclimation were examined. Three experiments were performed. (1) Freshwater (FW) to 30 salinity seawater (SW): plasma osmolality and chloride rose violently immediately post-transfer. At 6 h, gill Na–K-ATPase activity began to increase but most fish died from excessive plasma osmolality and Cl. (2) FW to 20 salinity SW: plasma osmolality and chloride increased immediately post-transfer, but more slowly than in (1), and began to decrease at 24 h. However it was not until 12 h post-transfer that gill Na–K-ATPase activity rose slowly. (3) FW to 20 salinity SW for 24 h, then to 30 salinity SW: after transfer to 30 salinity, plasma osmolality and chloride showed only a small increase initially then declined, while gill Na–K-ATPase activity started to rise rapidly within 3 h. The present results coincided with our previous morphological data concerning the ultrastructural responses of gill chloride cells. These are discussed to elucidate the osmoregulation mechanisms in tilapia during seawater acclimation.     

Relationship between phytoplankton cell size and the rate of orthophosphate uptake: in situ observations of an estuarine population

Orthophosphate uptake by a natural estuarine phytoplankton population was estimated using two methods: (1) ³²P uptake experiments in which filters of different pore sizes were used to separate plankton size-fractions; (2) ³³P autoradiography of phytoplankton cells. Results of the first method showed that plankton cells larger than 5 m were responsible for 2% of the total orthophosphate uptake rate. 98% of the total uptake rate occurred in plankton composed mostly of bacteria, which passed the 5 m screen and were retained by the 0.45 m pore-size filter. There was no orthophosphate absorption by particulates in a biologically inhibited control containing iodoacetic acid. Orthophosphate uptake rates of individual phytoplankton species were obtained using ³³P autoradiography. The sum of these individual rates was very close to the estimated rate of uptake by particulates larger than 5 m in the ³²P labelling experiment. Generally, smaller cells were found to have a faster uptake rate per m³ biomass than larger cells. Although the nannoplankton constituted only about 21% of the total algal biomass, the rate of phosphate uptake by the nannoplankton was 75% of the total phytoplankton uptake rate. Results of the plankton autoradiography showed that the phosphate uptake rate per unit biomass is a power function of the surface: volume ratio of a cell; the relationship is expressed by the equation Y=2x10^-11 X ^1.7, where Y is gP m^-3 h^-1 and X is the surface: volume ratio. These results lend support to the hypothesis that smaller cells have a competitive advantage by having faster nutrient uptake rates.     

Metabolic diversity in epibiotic microflora associated with the Pompeii wormsAlvinella pompejana andA. caudata (Polychaetae: Annelida) from deep-sea hydrothermal vents

Specimens of alvinellid polychaetes (Alvinella pompejana Desbruyères and Laubier, 1980 andA. caudata Desbruyères and Laubier, 1986) and their tubes were sampled from deep-sea hydrothermal vents at 13°N from the manned submersible Nautile during the Hydronaut cruise (October to November 1987) on the East Pacific Rise. Samples were subjected to bacterial analysis aboard the mother ship Nadir to detect bacteria involved in the nitrogen and sulphur cycles, in non-specific heterotrophic processes, and displaying resistance to selected heavy metals. Cultures were incubated at different temperatures under atmospheric and in situ (250 atm) pressures. Bacterial growth was observed in enrichment cultures for most metabolic types screened. Heavy-metal-resistant bacteria were also detected in many samples. No filamentous bacterial form was observed in the cultures. The results demonstrate a high metabolic diversity in episymbiotic flora, indicating that the worm (A. pompejana orA. caudata), its tube and its epiflora represent a complex micro-ecosystem.     

Biological production of nitrite in seawater

The relative importance of 3 different sources for biological production of nitrite in seawater was studied. Decomposition of fecal pellets of the copepod Calanus helgolandicus (at a concentration of approximately 12 g-at N/l), in seawater medium, released small amounts of ammonia over a 6 week period. It nitrifying bacteria were added to the fecal pellets nitrite was barely detectable over the same period. Decomposition of phytoplankton (present at a concentration of about 8 g-at particulate plant N/l) with added heterotrophic bacteria, released moderate amounts of ammonia over a 12 week period. If the ammonia-oxidizing bacterium Nitrosocystis oceanus was added to the decomposing algae, nitrite was produced at a rate of 0.2 g-at N/l/week. Heterotrophic nitrification was not observed when 7 open-ocean bacteria were tested for their ability to oxidize ammonia. The diatom Skeletonema costatum, either non-starved or starved of nitrogen, produced nitrite when growing with 150 or 50 g-at NO ₂ ^- -N/l at a light intensity of about 0.01 ly/min. When nitrate in the medium was exhausted, S. costatum assimilated nitrite. If starved of vitamin B₁₂, both non-N-starved and N-starved cells of S. costatum produced nitrite in the medium with 150 g-at NO ₃ ^- -N/l. Nitrate was not exhausted and cell densities reached 2x10⁵/ml due to vitamin B₁₂ deficiency. If light intensity was reduced to 0.003 ly/min under otherwise similar conditions, cells did not grow due to insufficient light, and nitrite was not produced. In the sea, it appears that, in certain micro-environments, decomposition of particulate matter releases ammonia with its subsequent oxidation to nitrite. The amounts of these nutrients and the rate at which they are produced are dependent upon the nature of the materials undergoing decomposition and the associated bacteria. In certain other areas of the sea, where phytoplankton standing stock is high and nitrate is non-limiting, excretion by these organisms is a major source of nitrite.     

Accumulation of metals by marine picoplankton

Pumping rates in undisturbed suspension-feeding bivalves were measured with a thermistor microflowmeter. Pumping rates are related to the dry weight of Clinocardium nuttallii, Macoma nasuta, Mytilus californianus and Chlamys hastata as the following equations: P=0.85 W^0.84, P=0.23 W^0.855, P=2.216 W^0.715, and P=2.419 W^0.943 respectively. Pumping rate and gill area increase at the same rate in C. nuttallii, M. californianus and C. hastata. The pumping rate per unit gill area is highest for C. nuttallii (approximately 0.14 cm³/s/cm²), with intermediate values for M. californianus (0.08 cm³/s/cm²) and C. hastata (0.11 cm³/s/cm²); the lowest rate was found in the deposit-feeder M. nasuta (0.0057 to 0.0089 cm³/s/cm²), which also has the smallest gill to body weight. C. nuttallii has the relatively smallest gill of the suspension feeders and the highest pumping rate per unit gill area. The absolute pumping rates, however, were highest for the filibranchs M. californianus and C. hastata. Indications are given that these differences are the results of different fluid-mechanical/ecological strategies to minimize the cost-benefit ratio.Contribution No. 132 of the Marine Science and Maritime Studies Center, Northeastern University     

Sclerotized protein in the shell matrix of a bivalve mollusc

The epsilon-amino groups of lysine and phenolic groups of tyrosine are most heavily concentrated in the newlydeposited organic matrix of the shell of the bivalve Mercenaria mercenaria. A phenoloxidase enzyme which oxidizes L-dihydroxyphenylalanine is present only in this new area of the shell matrix. Scanning electron micrographs of calcified secretions of the shell show that accretion lines, thought to be layers of organic matrix separating diurnal acceretions of calcium carbonate, are not developed until up to 4 d after deposition of shell material. These results suggest that the shell matrix is hardened by some kind of polymerization, and that lysine and tyrosine residues in the matrix are involved in the process. Accretion lines in polished and etched sections become visible only after complete hardening of the polymer occurs.     

Allozyme and colour differences between two sibling species of the heteronemertean Lineus torquatus from the Sea of Japan

Brown and cherry colour morphs of the heteronemertean Lineus torquatus Coe, 1901 from the Vostok Bay of the Sea of Japan were compared using allele frequencies at 18 isozyme loci. These morphs shared no common alleles at 9 of the 18 loci. The observed genetic differences are strong evidence that the brown and cherry colour morphs, which occur sympatrically, are different species. Nei's genetic distance between these species (D=1.135) is considerably higher than typical D values for congeneric animal species. As with much previous work on nemerteans, intraspecific allozymic variability in both species was high, with mean expected heterozygosity values 0.286±0.049 and 0.159±0.051 for L. torquatus (brown) and L. torquatus (cherry) species, respectively.     

Association between the mollusc bivalve Loripes lucinalis and a Chlamydia-like organism,with comments on its pathogenic impact,life cycle and possible mode of transmission

Loripes lucinalis is a littoral bivalve which has already been confirmed to harbour endo-cellular sulfur-oxidizing bacteria within its gills. Examination of the digestive gland of L. lucinalis collected from the Moulin Blanc Beach in the Bay of Brest (Brittany, France) revealed the existence of an additional association involving a Chlamydia-like organism. Three different forms of Chlamydia-like bacteria were observed: reticulate rod-shaped cells, electron-dense cells and enlarged cells. The reticulate rod-shaped cells and the electron-dense bodies are thought to represent the germinal initial body and infectious form of the bacteria, respectively. The enlarged cells were always associated with what are believed to be spherical or icosahedral phages. Initial infestation seems to occur by phagocytosis at the apical pole of the digestive cells of the tubule and duct epithelia. Within the host cell, the bacteria undergo binary fission and budding, forming an inclusion which gradually fills up the cell. Inclusions are generally between 15 and 30 m in size, and > 85% of all individuals examined possessed inclusion bodies. The level of infestation varied between individuals, some being heavily colonized, but did not seem to be related to season. Histological and ultrastructural observations suggest that, once developed, the colony has three possible fates: (1) the cells will degenerate due to phage infection; (2) colony overcrowding will occur, causing the development of electron-dense bodies that will be released into the lumen; (3) the entire membrane-bound inclusion will be released into the lumen and subsequently into the pallial cavity. Inclusions within the pallial cavity may be ingested by the host or may even be phagocytized by bacteriocyte cells of the gill. It is proposed that this association could be a form of symbiosis and that L. lucinalis may, therefore, be a rare example of an organism adapted to harbour two very different symbioses.     

Discussion of methods of investigating the food of fishes,with reference to a preliminary study of the prey of Gobiusculus flavescens (Gobiidae)

The validity, reliability and practicability of different methods for the investigation of stomach contents of a small, plankton-eating fish are discussed. Tests demonstrated the problems related to mass and volume measurements of planktonic food. Several nutrition indices, particularly Hynes's frequency of occurrence are criticized. A logarithmic version of Shorigin's index is proposed as a replacement for Ivlev's food selection index. A preliminary study on the food of Gobiusculus flavescens (Fabricius), sampled in July, 1975 at Helgoland, provides some numerical information on the composition of its stomach contents and food preference.     

Na<Superscript>+</Superscript>/K<Superscript>+</Superscript>–ATPase activity and gill ultrastructure in the hyper-hypo-regulating crab<Emphasis Type="Italic"> Chasmagnathus granulatus</Emphasis> acclimated to dilute,normal, and concentrated seawater

We studied Na⁺/K⁺ –ATPase activity and ultrastructure in gills of the hyper-hypo-regulating crab Chasmagnathus granulatus Dana, 1851 acclimated to different salinities: 10, 30 and 45, known to be hypo-, iso-, and hyper-osmotic to the hemolymph, respectively. After centrifugation of homogenates at 11,000 g, Na⁺/K⁺–ATPase activity was almost entirely found in the pellets from the posterior (6–8) and anterior (3–5) gills, whereas very little was detected in the supernatant liquid. Specific activity of gill 6 was 41.3, 30.2, and 28.2 µmol Pi h^–1 mg prot^–1 for crabs acclimated to 10, 30, and 45, respectively, the result for 10 being significantly higher than those at 30 and 45. Although the concentration of sodium at which the reaction rate is half-maximal (K _M) was similar in the three acclimation salinities, only the enzyme from crabs acclimated to 10 was inhibited by high sodium concentration. Specific activity of gill 5 increased with the increment in external salinity (10.1, 15, and 18.1 µmol Pi h^–1 mg prot^–1 for 10, 30, and 45, respectively), the only significant difference being that between the extreme salinities. The epithelium thickness of the dorsal portion of gill 6 showed a variation among salinities: 21.7, 15.8 and 17.2 µm for 10, 30 and 45, respectively. There were significant differences in epithelium thickness between the 10 and the other salinities. In all three salinities, the ultrastructure of gill 6 epithelium showed a high density of mitochondria, estimated by their volume fraction (Vv _m=0.307–0.355). These mitochondria were packed between extensive basolateral membrane interdigitations in ionocytes and pillar cells. Gill 5 showed three cell types: pillars which possess mitochondria packed between membrane folds only in their interdigitations with neighbouring cells; type-I cells 8.0 µm thick with low density of mitochondria (Vv _m=0.088), and type-II cells, 9.9 µm thick and rich in mitochondria (Vv _m=0.423), but lacking basolateral interdigitations. Vv _m of type-I cells of gill 5 was significantly lower than those of type-II cells of the same gill and the ionocytes of gill 6. No significant difference in Vv _m was detected between the latter cell types.Communicated by P.W. Sammarco, Chauvin     

The structure of the latero-frontal cirri on the gills of certain lamellibranch molluscs and their role in suspension feeding

Photomicroscopy and scanning electron microscopy were used to determine the fine structure of the latero-frontal cirri on the gills of three species of bivalve mollusc: Mytilus edulis Linn., Barnea candida (Linn.), and Petricola pholadiformis Lam. The component cilia are seen to branch off alternately along the length of each cirrus. Each of these free, lateral lengths of the cilia are separated from the next on their side by a mean distance of 0.6 m. They extend to a mean length of 2.7 m in M. edulis, the species described in detail. It is shown that a complete straining mesh can be formed over the gill ostium during normal filtering and the aperture of this mesh correlates well with the previously recorded filtering efficiency for Mytilus.     

On the feeding of Branchiostoma senegalense (Acrania: Branchiostomidae)

The feeding biology of Branchiostoma senegalense Webb from the North-West African shelf region off Mauritania was investigated. The hindgut contained almost exclusively seston components, ranging in size from less than 1 m (bacteria size) to 300 m. Numerous chloroplasts of ingested phytoplankton appeared to be undigested. Dunaliella salina was still alive when leaving the anus after 1 to 2 h. Detritus formed the greater part of the gut contents. The authors conclude that the lancelets feed mainly on detritus and thus are indiscriminate suspension feeders.     

Effect of daylight variations on the energy budgets of shallow-water corals

Energy budgets were determined for small pieces (nubbins) of the coralsPocillopora damicornis, Montipora verrucosa andPorites lobata living at a water depth of 3 m on the fringing reef of Coconut Island, Kaneohe, Hawaii. The budgets were determined for three different types of day: an ideal day with no cloud and an in situ daily integrated irradiance at 3 m of 14.385 E m^–2 d^–1; a normal day with sporadic cloud cover and daily irradiance of 11.915 E m^–2 d^–1; and an overcast day with daily irradiance of 6.128 E m^–2 d^–1. On the ideal day, the energy fixed in photosynthesis was more than that required for respiration and growth of both zooxanthellae and animal components of the association, and there was a predicted loss of between 19.3 and 32.4% of the energy fixed. On a normal day, the total photosynthetic energy fixation was lower and the excess was between 12.1 and 27.9% of the energy fixed. On the overcast day, however, in bothPocillopora damicornis andPorites lobata energy expenditure exceeded photosynthetic energy fixation and the budget was in deficit. Estimates of rate of mucus secretion on an overcast day were derived and, when incorporated into the energy budget, it was predicted that all three species would have a deficit budget, necessitating the catabolism of lipid reserves. From published values for lipid storage in these species it was calculated that the reserves would last from 28 d inPocillopora damicornis to 114 d inM. verrucosa. A model is suggested in which corals draw upon their extensive lipid stores on days of sub-optimal light, replenishing the reserves again when daily light levels are high, and finally excreting the excess energy fixed, as mucus-lipid when the lipid stores are replete.     

Particle capture in the musselMytilus edulis: The role of latero-frontal cirri

Microscope video graphs of particle paths near one-filament-thick mussel gill preparations, stimulated with a nerve transmitter (10^–6 M serotonin which restores normal ciliary activity), were used to disclose the capture of 6 m algal cells. Suspended algal cells carried with the water were stopped for a while at the entrance to the interfilament gap by the action of the latero-frontal cirri (Ifc), and transferred to the frontal side of the filament to be transported towards the marginal food groove. The event of transfer took place during approximately a time interval of 1150 to 1/25 s. To gain a better understanding of the capture mechanism and retention efficiency versus particle size, the flow through and around the Ifc was theoretically estimated. Normally beating Ifc create periodic, unsteady, three-dimensional flows at the entrance to the interfilament canal. During the active beat most of the water is deflected to flow around the branching cilia of the Ifc while some of the water is strained by these. Large particles (> 4 m) are stopped and transferred to the frontal current, whereas smaller particles either follow the flow around the Ifc and escape or they are stopped by the branching cilia.     

Does displacement of spawning time occur in the sibling species Cerastoderma edule and C. lamarcki?

The spawning cycles of the sibling species Cerastoderma edule (L.) and C. lamarcki (Reeve) were studied in allopatric and sympatric Danish populations from 1978–1980. The results do not support earlier findings of reproductive character displacement in sympatric populations of the two species. New records of hybrid formation and intermediate forms are presented and discussed.     

Unusual nutrition of the “Pompeii worm” Alvinella pompejana (polychaetous annelid) from a hydrothermal vent environment: SEM,TEM, 13C and 15N evidence

Morphological and histological studies of Alvinella pompejana (a polychaete living in the vicinity of hydrothermal vents of the East Pacific Ocean) were performed using light, scanning and transmission electron microscopy. The worms were collected in April–May 1979, during the Rise cruise by the submersible Alvin on the crest of East Pacific Rise at 21°N. The digestive tracts contained many sulfide particles (as determined by microprobe analysis) associated with organic matter and bacteria. Bacterial communities of different morphological types (cocci and filaments) were also observed at different levels of the worm's outer teguments. An atypical (possibly bacteria-derived) nutritional source of carbon and nitrogen for A. pompejana is indicated by the natural abundances of ¹³C:¹²C and ¹⁵N:¹⁴N in its tissues.Contribution No. 807 from Centre Océanologique de Bretagne, B.P. 337, F-29273 Brest Cédex, France