Synergistic effects of lead,salinity and temperature on embryonic development of the mussel Mytilus galloprovincialis

Combined effects of lead, salinity and temperature on the embryonic development of the mussel Mytilus galloprovincialis Lmk. were studied under laboratory conditions. The basic experimental design was a 4x6 factorial experiment using 4 lead concentrations (100, 250, 500 and 1000 ppb Pb²⁺) and 6 salinity levels (from 25 to 37.5 with 2.5 intervals). These factorial designs were carried out at three constant temperatures (150, 17.50 and 20°C). The statistical analysis indicated that salinity changes have more effect on the embryonic development than temperature. Optimal development was observed at 34.8 and 15.6°C, which is in accordance with observations in the field. The effect of lead was mininal in optimal salinity and temperature conditions. The deleterious effect of lead on the embryonic development was especially conspicuous at 20°C. Since in nature spawning occurs at temperatures inferior to 20°C, lead will probably not drastically decrease the potential recruitment of mussel spat in the littoral populations of the northern Adriatic Sea, where the salinity of the water is relatively stable. Under experimental conditions, lead caused a delay or inhibition of the embryonic development with the occurrence of a large number of abnormal larvae.     

The combined effects of salinity and temperature on larvae of the mussel Mytilus edulis

The combined effects of salinity and temperature on survival and growth of larvae of the mussel Mytilus edulis (L.) were studied. The effects of salinity and temperature are significantly related only as the limits of tolerance of either factor are approached. Survival of larvae at salinities from 15 to 40 is uniformly good (70% or better) at temperatures from 5° to 20°C, but is reduced drastically at 25 °C, particularly at high (40) and low (20) salinities. Larval growth is rapid at a temperature of 15 °C in salinities from 25 to 35, at 20 °C in salinities from 20 to 35. Optimum growth occurs at 20 °C in salinities from 25 to 30. Growth decreases both at 25° and 10 °C; the decline is most drastic at high (40) and low (20) salinities.Part of a study completed at the Bureau of Commercial Fisheries, Biological Laboratory, Milford, Connecticut, USA, while on a UNESCO Fellowship.     

Comparative effects of two antifouling paints on the oyster Crassostrea gigas

The effects of two antifouling paints, one containing organotin compounds, the other copper oxide, were studied from September 1983 to October 1984 on Crassostrea gigas (Thunberg) grown under natural field conditions in the Bay of Arcachon, France. The organotin paint reduced growth rate expressed as weight, length and width, but did not affect shell height; it drastically decreased the dry condition factor and shell density, but did not affect the viability of embryos and larvae from exposed oysters. However, some decrease in larval growth rate was observed. The copper paint had no effect on oyster growth, but lowered the condition factor compared to controls. Neither viability of embryos or larvae nor larval growth were affected by this paint.     

Effects of artificial eutrophication on the metabolism of the Japanese oyster Crassostrea gigas

The morphology of the primary tube feet in 15 species of comatulid (unstalked) crinoids from coral reefs in the Palau Islands and Lizard Island, Great Barrier Reef, was investigated using close-up underwater photographs of the tube feet taken in the natural habitat. Measurements of length of the tube feet and their spacing along the pinnule were taken from these photographs. Tube feet of species of the family Comasteridae have a mean length of 0.75 mm and a mean spacing of 6 tube feet mm^-1. Tube feet of non-comasterid species occurring in the same environments have a mean length of 0.55 mm and a mean spacing of about 8 tube feet mm^-1. The relationship between spacing and length of the tube feet is highty significant for both the Palau and Lizard Island samples (P<0.01). Species having longer and more widely spaced tube feet live partly concealed within the infrastructure of the reef and hold the arms and pinnules in a multidirectional posture. Species having shorter and more closely spaced tube feet perch on top of reef pinnacles or alcyonarians and form planar filtration fans normal to unidirectional currents or wave oscillations. Longer and more widely spaced tube feet in species dwelling within the reef infrastructure provide more efficient filtration in the slow, meandering flow prevailing there. Closer spacing of the tube feet in species exposed to near-mainstream flow provides a more efficient filtration mechanism at higher flow velocities. Reduced length of the tube feet in these species may be a consequence of closer spacing of the pinnules. Differentiation of these co-occurring species in spacing and length of the tube feet implies differentiation in food particles captured. This may in some cases constitute resource partitioning. Most species which overlap in living habits are significantly different in spacing and length of the tube feet.     

Interaction of genotype and salinity in larvae of the oyster Crassostrea virginica

Adult Crassostrea virginica were obtained from 4 populations and spawned in the laboratory. The larvae from the within-population crosses and the hybrid crosses were raised at 4 salinities. There were no significant differences in survival of the larvae between the populations. However, one set of hybrids did show overdominance in survival. There were genetic differences between the populations in growth rate, but the expression of the differences depended upon the salinity, i.e, there was significant genotype-environment interaction. There were expressions of nonadditive genetic effects in the hybrid crosses, but the direction and magnitude was dependent upon the salinity. There was as much difference between populations from the same estuary as there was between populations from geographically isolated populations.     

Effect of zinc on growth and development of larvae of the Pacific oyster Crassostrea gigas

Following the observation of periodic high concentrations of zinc in estuarine waters used in the White Fish Authority's oyster hatchery at Conway, North Wales, two beaker trials were conducted to study the effect of zinc, over the range recorded, on the young stages of larvae of Crassostrea gigas. Zinc, added to sea water both as zinc sulphate and as a natural mine-adit water, was applied for a period of 5 days, after which larvae were maintained for a further 5 days in sea water alone. Increasing concentrations over the range 125 to 500 g/l Zn resulted in decreasing growth, and increasing incidence of abnormality and larval mortality. A second trial with zinc sulphate showed 50 g/l Zn to have little effect on larval development, a progressive decrease in growth at 100 and 150 g/l, and no growth at 200 g/l. It is suggested that the deleterious effect of short-term exposure to zinc may well have contributed to the intermittent failure of larvae and irregular productivity previously recorded at the hatchery. It is also possible that zinc contamination in estuaries may affect natural oyster breeding, and may have to be considered in the future siting of hatcheries for seed production.     

Induction of metamorphosis in larvae of the oyster Crassostrea gigas using neuroactive compounds

Chemical (neuroactive compounds at different concentrations and exposure times) and physical (water agitation, light) factors with potential effects on the metamorphosis of larvae of the oyster Crassostrea gigas (Thunberg) larvae have been studied. The neurotransmitters l-dihydroxyphenylalanine (DO), epinephrine (EP), norepinephrine (NE), and acetylcholine (AC) have been identified as very active inducers of metamorphosis, whilst serotonin (SE), dopamine (DA) and potassium (K) were less effective inducers. The -aminobutyric acid (GA) and ammonium (AM) were found ineffective at the concentrations tested. Exposure to 10^-4 M EP for 15 min was sufficient to promote >80% metamorphosis within 48 h, whereas NE required 2 h to exert comparable induction. Maximum induction by DO (>50%) was achieved after 2 h exposure to 10^-4 M. However, unlike EP and NE, DO was lethal at that concentration in the long term. Maximum induction by AC (30% metamorphosis) was achieved at a concentration of 10^-4 M. In contrast to other neurotransmitters, AC induced settlement behaviour, cementation and eventual metamorphosis, yielding postlarvae which were all attached to the substratum. EP and NE triggered the morphogenetic changes, by-passing settlement and leading to postlarvae not cemented to the substratum. DO induced mostly attached spat at low concentrations (10^-5 M) and unattached spat at high concentrations (10^-4 M), and a similar pattern was apparent for the weaker inducers SE and DA. Regarding physical factors, a highly reflectant surface significantly increased the percentage of attached spat obtained, compared to a dark bottom. No consistent effect of water current or light was detected on the production of unattached spat. The three different forms of induction are discussed in relation to different regulatory pathways of settlement and metamorphosis.     

Effect of zinc on the settlement of the oyster Crassostrea gigas

The influence of zinc upon the pattern and success of settlement was examined in the oyster Crassostrea gigas. Late larvae were more tolerant of zinc than embryos. A delay in settlement was recorded in treatments containing zinc as low as 125 μg/l, and numbers of larvae settling was reduced in the presence of zinc. Larvae subjected to zinc immediately prior to settlement showed evidence of slowing in behavioural development. Those larvae which settled in the presence of zinc, however, when ongrown in clean water were as viable as controls. Zinc at concentrations of 250 and 500 μg/l suppressed spat growth, but recovery was rapid upon subsequent on-growing in clean water conditions.     

Metabolic fate of aromatic amines in the mussel Mytilus galloprovincialis

The postmitochondrial fraction of the digestive gland from the marine mussel Mytilus galloprovincialis possesses FAD-containing monooxygenase (EC 1.14.38) but lacks cytochrome P-450 dependent benzo(a)pyrene monooxygenase (EC 11.4.14.1). This is also evidenced by the ability of the mussel preparation to activate carcinogenic aromatic amines, but not carcinogenic benzo(a)pyrene, to Salmonella typhimurium TA 98 mutagens. This metabolic activity is NADPH dependent. Mussel digestive gland postmitochondrial fraction also possesses the enzymes needed for the detoxicating part of the aromatic amine metabolism: UDP-glucuronyl transferase (EC 2.4.1.17) and -glucuronidase (EC 3.2.1.31). Under the experimental conditions used here, this aromatic amine metabolic pathway converts up to 8% of 2-acetylamino(9-¹⁴C)fluorene, but not (G-³H)benzo(a)pyrene, to water soluble glucuronides. Glucuronic acid stimulates the formation of these glucuronides. The metabolites liberated from these glucuronides by the -glucuronidase treatment could be converted to TA 98 strain mutagens by the carp liver postmitochondrial fraction, but not by the mussel's digestive gland preparation. The presence of such a selective potential for the bioactivation and detoxication of aromatic amines, and not polycyclic aromatic hydrocarbons, in the marine invertebrate(s) may bring new insight to our understanding of the effects and the fate of carcinogens in the marine environment.     

Settlement behavior and metamorphosis of oyster larvae (Crassostrea gigas) in response to bacterial supernatants

Veliger larvae of the oysterCrassostrea gigas (Thunberg) responded to unknown dissolved chemical inducers found in supernatants of cultures of the bacteriaAlteromonas colwelliana andVibrio cholerae. The response, which was similar to that seen when larvae were exposed to the neurotransmitter precursor L-3,4-dihydroxyphenylalanine (L-DOPA), consisted of an initial settlement phase of swimming with the foot extended and crawling on the substrate. Subsequently larvae attached to the substrate and metamorphosed. The percentage of veligers metamorphosing following inducation of settlement behavior was higher in a group of older larvae, a response similar to that seen with L-DOPA, suggesting that competence to respond to bacterial supernatants is divided into two phases: behavioral competence followed by morphogenetic competence. Following size exclusion chromatography, the molecular weight of the peak containing the activity which induced settlement behavior was determined to be 300 daltons. Autoclaved Marine Broth, which induced low levels of settlement behavior also contained this low molecular weight active peak, suggesting that an oyster settlement inducer is also present in this medium.Contribution # 137 from the Center of Biotechnology, Marine Biotechnology Institute, University of Maryland, USA     

Factors affecting the flux of arsenic through the mussel Mytilus galloprovincialis

Radiotracer experiments were designed to study the effects of certain environmental and biological factors on arsenic accumulation and elimination processes in the Mediterranean mussel Mytilus galloprovincialis. Arsenic (as arsenate) uptake increased with increasing arsenic concentration in the water; however, the response was not proportional, indicating that accumulation was partially suppressed at higher external arsenic concentrations. In general, approximately 80% of the ⁷⁴As taken up was associated with the soft parts, with small mussels concentrating ⁷⁴As to a greater degree than larger individuals. The highest ⁷⁴As concentrations were recorded in the byssus and the digestive gland. Increased temperature enhanced both arsenic uptake and loss. Mussels in sea water at 19 S accumulated approximately three times more ⁷⁴As than those held at 38 S. Arsenic loss was much less affected by salinity, with only a tendency for greater arsenic retention noted at lower salinities. Studies carried out in the laboratory and in situ revealed that arsenic turnover was significantly more rapid in actively growing individuals living under natural conditions. Arsenic-74 loss from the in situ group was essentially biphasic, with biological half-times of approximately 3 and 32 days for the fast and slow compartments, respectively. The active secretion of arsenic in the byssal threads contributed to the total elimination of the element from the mussels.     

Effects of temporary starvation on the survival,and on subsequent feeding and growth,of oyster (Crassostrea gigas) larvae

Larvae of oysters, Crassostrea gigas, were maintained without food for 1 to 8 d after fertilization, and fed daily thereafter. There was little difference in survival and growth between controls and larvae kept without food for 2 or 3 d. Survival and growth rates were depressed in larvae starved for 4 or 5 d. For larvae starved for 6 to 8 d, survival was negligible or nil; even those larvae which survived the starvation period died later in the presence of food, apparently because of impaired digestion. Therefore, food availability in the first few days after spawning appears to be of paramount importance to the successful recruitment of Pacific oysters.     

Single and combined effects of heavy metals and hormones on lysosomes of haemolymph cells from the mussel Mytilus galloprovincialis

Effects of heavy metals on lysosomes were studied in living cells from the mussel (Mytilus galloprovincialis Lam.). Haemolymph cells were obtained from the mussel adductor muscle, stained with neutral red (NR), and analysed by digital imaging to evaluate NR retention times within lysosomes. Exposure to Hg²⁺, Cd²⁺ and Cu²⁺ induced a reduction of NR retention time, indicating lysosomal membrane destabilisation. The intensity of these effects was correlated with the metal affinity for sulfhydryls. In contrast, Zn²⁺ showed no effect on lysosomes. Moreover, 200 μM Zn²⁺ protected lysosomes against the effects of Cd²⁺ and Cu²⁺, but not against Hg²⁺. Cell loading with the fluorescent pH probe Lyso Sensor followed by digital imaging showed a rise of lysosomal pH induced by Cd²⁺ and Hg²⁺, while Zn²⁺ prevented the effect of Cd²⁺ and also partially that of Hg²⁺. The different protective effect of Zn²⁺ against Hg²⁺ suggests a dual action of Hg²⁺ on lysosomes, possibly involving both membrane destabilisation and proton pump inhibition. Cell exposure to 17 β-estradiol also caused a reduction of NR retention time, which was synergistic to that of Hg²⁺. This suggests a common pathway between metals and hormone, possibly involving Ca²⁺ signaling. Received: 17 November 1999 / Accepted: 29 June 2000     

Stylochus mediterraneus (Turbellaria: Polycladida), predator on the mussel Mytilus galloprovincialis

The polyclad Stylochus mediterraneus Galleni has been found associated with the mussel Mytilus galloprovincialis Lmk., on which it feeds. Polyclads allowed to feed freely on groups of mussels of different sizes preyed mainly on small mussels <25 mm in length. The predation rate (number of mussels eaten per no. of polyclads per no. of days) ranged between 0.07 and 0.33. The average amount of food ingested by 1 polyclad feeding on mussels 16 to 25 mm or 26 to 35 mm in length was 7.1 and 11.2 (dry weight) mg per day, respectively. Before penetrating the mussels, the worm first straddles the valves at the posterior edge of the shell and then, after having digested the posterior adductor muscle, removes and swallows the soft parts of the prey.     

Oil pollution monitoring in the Lagoon of Venice using the mussel Mytilus galloprovincialis

Gas chromatographic analyses of mussels, Mytilus galloprovincialis, from different areas of the Lagoon of Venice show that these organisms contain a very complex mixture of hydrocarbons attributable to fuel oil contamination. The measured amounts normally range from 0.8 to 8.7 mg/100 g wet weight, but values as high as 22.0 mg/100 g have been recorded. This high value indicates a saturation limit for these organisms which is considerably higher than those values normally found in mussels from the lagoon. The aliphatic hydrocarbon levels in mussels are related to distance from pollution sources and to the degree of exchange between the sea and the area sampled. On the basis of this relationship between overall hydrocarbon pollution load and level of contamination of M. galloprovincialis, it appears that this bivalve might be effectively utilized as a self-integrating monitoring index of oil pollution in the wasters of the lagoon.     

Effects of monospecific algal diets of varying biochemical composition on the growth and survival of Pacific oyster (Crassostrea gigas) larvae

Using monospecific diets of Thalassiosira pseudonana cells grown under different steady-state conditions, it was determined that higher growth rates of larval Crassostrea gigas Thunberg were obtained when fed T. pseudonana cells grown under high light. High light grown T. pseudonana cells consistently contained relatively more of the saturated fatty acids 14:0 and 16:0. Considered over three independent experiments, high light grown T. pseudonana cells were lower in protein and higher in carbohydrate than low light grown cells. Higher growth rates of larval C. gigas were obtained on diets with more of the essential fatty acid (EFA) 22:63, and less of the other EFA, 20:53. The relative requirements of C. gigas larvae for the essential fatty acids 20:53 and 22:63 are discussed. Faster growing larvae contained higher percentages of the fatty acids 14:0 and 16:0, and lower percentages of 22:2j. Oyster growth rates were correlated with their content of the fatty acids: 14:0, 16:0 and 22:2j in two experiments utilizing separately spawned batches of larvae. Fatty acid profiles are proposed as a technique for assessing larval condition. C. gigas larvae contained ten times the percent composition of the FAs 16:43, 18:17, 20:17 and 22:2j compared with their diet. Correlation analysis suggests that the dietary source of 18:17, 20:17 and 22:2j was 16:17. It is concluded that T. pseudonana cells grown under high light are a superior diet for C. gigas larvae in comparison with low light grown cells of the same species.     

Combined effects of temperature and salinity on embryos and larvae of the northern bay scallop Argopecten irradians irradians

The combined effects of temperature and salinity on embryonic development and on larval survival and growth to setting size of the northerm bay scallop Argopecten irradians irradians (Lamarck) were studied in the laboratory. A 6x6 complete factorial design was used; temperatures ranged from 10° to 35°C, at 5C° intervals, and salinities ranged from 10 to 35S, at 5S intervals. Response-surface contour diagrams were generated to provide estimates of conditions for optimal responses. Normal development of embryos occurred over a very narrow range of temperature and salinity. Survival of larvae occurred over a wider range of temperature and salinity than did embryonic development or growth of larvae. Satisfactory growth (>70% of the maximum observed value) occurred only at high temperature-high salinity conditions; optimal conditions for survival occurred at similar salinities, but at slightly lower temperatures. Temperatures of 35°C or greater and/or salinities of 10S or less were lethal for all life stages studied. Both salinity and temperature exerted significant effects on development and survival, but temperature was clearly the dominant factor influencing growth. It is suggested that northern bay scallop embryos and larvae be reared at their respective optimal temperature-salinity levels so as to increase efficiency of aquaculture operations.This paper is adapted from a thesis submitted to the College of Fisheries, University of Washington, in partial fulfillment of the requirements for the MS degree. This study was conducted at the NMFS Laboratory in Milford, Connecticut, USA     

Purification and immunolocalization of a vitellin-like protein from the Pacific oyster Crassostrea gigas

A female-specific protein from the hemolymph, and a related ovarian protein were identified in the Pacific oyster Crassostrea gigas using immunological procedures. The latter protein was isolated from ovarian extract by a combination of precipitation in distilled water, ammonium sulfate fractionation, hydroxylapatite chromatography and gel filtration on a Sepharose 6B column, and was identified as a macromolecular glycolipoprotein of 500 kD; sodium dodecylsulfate polyacrylamide gel electrophoresis, under reducing conditions, revealed seven heterogeneous polypeptides. Using a specific antiserum against purified protein, we also examined the immunohistochemical distribution of the protein at light- and electron-microscopic levels. Light microscopy revealed its presence in the oocytes, and electron microscopy identified it in yolk granules and the vitelline coat of the oocytes. These results indicate that the purified protein is a vitellin-like protein. Since no organ other than the ovary reacted with the antiserum, it is probable that the protein is produced inside the ovary, probably autosynthetically by oocytes.     

Deep-sea caging of the mussel Mytilus galloprovincialis: Potential application in ecotoxicological studies

We experimented with caging the Mediterranean mussel (Mytilus galloprovincialis) at various depths for 69 d to measure basic physiological parameters, histological response and bio-accumulation of contaminants in a deep-sea contaminated area. In preliminary experiments, we demonstrated, under artificial pressure conditions, the ability of mussels Mytilus galloprovincialis to tolerate rapid immersion (at a speed of up to 120 m min^?1). In situ experiments were performed using submerged lines enabling mussels to be maintained at depths ranging of 40–1550 m with survival rates ranging from 80 to 38%, respectively. No significant differences in condition indexes were observed between treated and control specimens. However, histological observations demonstrated a clear reduction in thickness of the digestive epithelium with increasing depth exposure. By determining the contaminants in caged mussels, we found the following values for chromium accumulation: 27.4 μg g^?1 dry weight at 580 m depth and 9.8 μg g^?1 dry weight at 1550 m. Selected stations were located downstream of an industrial effluent at 420 m. The biological and environmental consequences of deep-sea contamination demonstrate the suitability of caged mussels for monitoring contaminant accumulation.