Toxic metal health risk by mussel consumption

Seafood is a major dietary food worldwide. However, seafood consumption by humans can induce health risk because seafood may be contaminated by various pollutants. The mussel Mytilus galloprovincialis is widely distributed in the coastal waters of Montenegro, SouthEast Adriatic Sea. Here, Zn, Fe, Cu, Ni, Cd, Pb, As, and Hg contents in M. galloprovincialis from ten sites were analyzed to investigate health risks associated with the consumption of wild and cultivated mussels. Since there is a lack of data on the mussel consumption rate in Montenegro, the amount of mussels that can be ingested weekly over a lifetime with no risk of negative health effects was calculated using provisional tolerable weekly intakes (PTWI). We found that Cd concentrations were the limiting factor for mussels as a food. The weekly consumptions of 0.64–1.2 kg of fresh wild and 0.84–1.2 kg of fresh cultivated mussel would be sufficient to reach the PTWI_Cd, which may result in a risky weekly intake of Cd for long-term exposure. Moreover, weekly intake of 125 g mussels was used to calculate the dietary intake of trace elements by mussel consumption and compared with the prescribed PTWIs. Here, we found that there is no risk for human health for all investigated elements. In this case, the highest Cd level obtained in wild and in cultivated mussels represents 19.8 and 14.9% of the PTWI_Cd, respectively. This is the first study in Montenegro giving an assessment of the health risk from trace elements via the consumption of wild and cultivated M. galloprovincialis.     

Response of mussel Brachidontes variabilis to chlorination

Brachidontes variabilis is a common fouling mussel species in cooling water systems of tropical coastal power stations. However, there are hardly any data available on the response of B. variabilis to chlorine, a commonly used antifouling biocide. Therefore, lethal and sublethal responses of this mussel to chlorine are of considerable interest to the industry. The response of mussels in terms of mortality pattern (LT₅₀ and LT₁₀₀) and physiological activities (oxygen consumption, filtration rate, foot activity and byssus thread production) in different size groups (with shell lengths of 7–24 mm) of B. variabilis was studied in the laboratory under different chlorine concentrations (0.25, 0.50, 0.75 and 1.00 mg l^?1 for sublethal responses and 1, 2, 3 and 5 mg l^?1 for mortality). The results showed that the exposure time for 100% mortality of mussels decreased significantly with increasing chlorine concentration. However, mussel size was not a determinant of its chlorine tolerance: all size groups tested (with shell lengths of 7–24 mm) took comparable exposure times to reach 100% mortality at a given chlorine concentration (1–5 mg l^?1). All size groups of B. variabilis showed a progressive reduction in physiological activities such as oxygen consumption, filtration rate, foot activity and byssus thread production, when chlorine residuals were increased from 0 to 1 mg l^?1. The data generated in the present work are compared with similar data available for other tropical fouling mussel species to see how far relative chlorine toxicity could have influenced the relative distribution of the mussels inside the seawater intake tunnel of a power station at Kalpakkam in India. It is shown that in this insufficiently chlorinated system, the relative distribution of Brachidontes striatulus, B. variabilis and Modiolus philippinarum reflects the relative tolerance of the species to chlorine.     

The effect of cadmium on antioxidant responses and the susceptibility to oxidative stress in the hydrothermal vent mussel Bathymodiolus azoricus

Hydrothermal vents are a unique environment of extreme physical–chemical characteristics and biological species composition. Cd is a toxic non-essential metal present in high concentrations in the hydrothermal vent environment, contrary to those found in marine coastal areas. Cd toxicity has been related, among other things, with reactive oxygen species production, even though this is a non-redox metal. Bathymodiolus azoricus is a deep-sea Mytilid bivalve very common in the Mid Atlantic Ridge (MAR) hydrothermal vent fields and very little is known about the antioxidant defence system in this specie. Because lethal Cd concentration in B. azoricus is unknown, the aim of this study was to assess the effects of a Cd concentration higher than that found in the hydrothermal vents on oxidative stress biomarkers, such as antioxidant enzymes. Mussels were exposed to 100 μg l⁻¹ Cd during 24, 48 and 144 h, respectively, in a pressurized aquarium (IPOCAMP). Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidases (GPx), total oxyradical scavenging capacity (TOSC), metallothionein (MT) and lipid peroxidation (LPO) were measured in the gills and mantle of B. azoricus. The results indicate that gills are first affected by Cd toxicity. This may be due to different physiological functions of the tissues and by the presence of thio and methanotrophic symbiotic bacteria in the gills. The SOD and CAT are inhibited during the first day of exposure in the gills, although TOSC and MT concentrations were the same in control and exposed mussels. In the mantle, enzymatic activation only occurred after 6 days, and no significant differences in MT concentrations were found in the control and exposed mussels during the first day, as observed in the gills.     

Potential effects of a non-indigenous predator in its expanded range: assessing green crab, <Emphasis Type="Italic">Carcinus maenas</Emphasis>, prey preference in a productive coastal area of Atlantic Canada

The non-indigenous green crab (Carcinus maenas) is an important predator on bivalve wild beds in coastal areas worldwide. This study explored size-dependent green crab prey preference on American oysters (Crassostrea virginica), blue mussels (Mytilus edulis), and soft-shell clams (Mya arenaria) in a productive coastal system of Atlantic Canada. Using two sizes of prey and three different experimental manipulations, small, medium, and large green crabs were given a choice among these three bivalves, and their daily feeding rates were monitored over the course of 3 days. For both prey sizes, green crabs showed an early feeding preference for soft-shell clams and, only as they declined in numbers, a switch toward mussels and subsequently toward oysters. We found that such changes in the timing (order) of prey preference are related to prey differences in shell thickness, a fairly reliable indicator of prey shell strength.     

Simultaneous, long-term monitoring of valve and cardiac activity in the blue mussel Mytilus edulis exposed to copper

Valve and cardiac activity were simultaneously measured in the blue mussel (Mytilus edulis) in response to 10?d copper exposure. Valve movements, heart rates and heart-rate variability were obtained non-invasively using a Musselmonitor^® (valve activity) and a modified version of the Computer-Aided Physiological Monitoring system (CAPMON; cardiac activity). After 2?d exposure of mussels (4 individuals per treatment group) to a range of dissolved copper concentrations (0 to 12.5?M as CuCl₂) median valve positions (% open) and median heart rates (beats per minute) declined as a function of copper concentration. Heart-rate variability (coefficient of variation for interpulse durations) rose in a concentration-dependent manner. The 48?h EC₅₀ values (concentrations of copper causing 50% change) for valve positions, heart rates and heart-rate variability were 2.1, 0.8, and 0.06?M, respectively. Valve activity was weakly correlated with both heart rate (r?=?0.48?±?0.02) and heart-rate variability (r?=?0.32?±?0.06) for control individuals (0?M Cu²⁺). This resulted from a number of short enclosure events that did not coincide with a change in cardiac activity. Exposure of mussels to increasing copper concentrations (≥0.8?M) progressively reduced the correlation between valve activity and heart rates (r?=?0 for individuals dosed with ≥6.3?M Cu²⁺), while correlations between valve activity and heart-rate variability were unaffected. The poor correlations resulted from periods of valve flapping that were not mimicked by similar fluctuations in heart rate or heart-rate variability. The data suggest that the copper-induced bradycardia observed in mussels is not a consequence of prolonged valve closure.     

The territorial flight of male houseflies (Fannia canicularis L.)

Summary The heart rate (HR) of three male and five female European blackbirds (Turdus merula) was monitored by radiotelemetry under three conditions: in a dark cage, in a lighted cage, and in an outdoor aviary. In all three, the response to recorded bird song was tested.The resting HR in the cage ranged from 4.8 to 6.3 beats per second. The HR of all the birds changed in response to playback of conspecific song as well as of the songs of other species. In 68% of trials a typical biphasic HR curve was obtained, with acceleration followed by deceleration (Fig. 2). The time from stimulus onset to the second curve inflection (t _iII) averaged 15–18 s; this was significantly longer for conspecific than for heterospecific song. The parameter t _iII was used as a basic measure of the response.The reaction of the males was longer-lasting than that of the females. The possibility that this reflects the territorial role of the songs is discussed. Response durations were longest in the dark cage, and shortest in the aviary. The reason is thought to be the gradation in total stimulus input available to the bird.The results suggest that the HR response consists of an unspecific component (acceleration) plus a component specific to bird sounds (deceleration). At this fundamental level of stimulus processing it is possible to study reactions to single strophes not only in males, as with more traditional methods, but also in females and young birds.     

Depression of feeding and growth rates of the seastar Evasterias troschelii during long-term exposure to the water-soluble fraction of crude oil

To test the effect of petroleum hydrocarbons on predation by the seastar Evasterias troschelii (Stimpson, 1862) on the mussel Mytilus edulis (L.), we exposed the predator with the prey to six concentrations of the water-soluble fraction (WSF) of Cook Inlet crude oil. Seastars and mussels were collected at Auke Bay, Alaska, in November 1980. During a 28 d exposure in a flow-through system, seastars were more sensitive to the WSF than mussels: the LC₅₀ for the seastars was 0.82 ppm at Day 19 and, although no mussels were exposed to WSF for more than 12 d, none died. Daily feeding rates (whether in terms of number of mussels seastar^-1 d^-1 or dry weight of mussels seastar^-1 d^-1) were significantly reduced at all concentrations above 0.12 ppm. At 0.20, 0.28 and 0.72 ppm WSF, daily feeding rates (in terms of dry weight of mussels) were, respectively, 53, 37, and 5% of the control rate; at the two highest concentrations (0.97 and 1.31 ppm WSF), the seastars did not feed. Seastars at concentrations greater than 0.12 ppm WSF grew slower than individuals from the control group and the 0.12 ppm-treatment group combined. These laboratory results show that E. troschelii is more sensitive to chronic low levels of the WSF of crude oil. The possibility that such oil pollution could reduce predation and permit M. edulis to monopolize the low intertidal zone of southern Alaska remains to be studied.     

Assimilation of trace elements by the mussel Mytilus edulis: effects of diatom chemical composition

Mussels have been widely used as bioindicators of coastal contamination, and recent reports have demonstrated that metals are accumulated from both the dissolved phase and from ingested food. In the winter and spring of 1995, we examined the influence of the chemical composition of food (protein content, trace element concentrations and ratios in the diatom Thalassiosira pseudomana) on the assimilation of six trace elements (Ag, Am, Cd, Co, Se and Zn) in the mussel Mytilus edulis (L.). Differences of up to 38% in diatom protein content had no major influence on the assimilation of any trace element or carbon. Protein assimilation in M. edulis examined with a ³⁵S radiotracer was also independent of protein content in the diatoms. Similarly, Se assimilation in mussels was not affected by the different Se concentrations in the diatoms. Cd assimilation increased with increasing Cd concentration, presumably due to higher desorption of Cd under acidic conditions typical of the mussel gut. Zn assimilation was inversely related to Zn concentration in the food particles, implying a partial regulation of this metal in the mussels. There was no evidence of any interaction of Cd and Zn in their assimilation by the mussels. These results suggest that mussels are highly responsive, in an element-specific way, to some components of ingested food (e.g., metal concentration), but other food components (such as the biochemical composition of the algae) have little effect on assimilation.     

Increased metabolic cost of maintenance in hybrids of the stone crabs Menippe mercenaria and M. adina

Three recently emerging genotype classes of the genus Menippe (M. mercenaria, M. adina, and their hybrids) were examined for potential physiological correlates of gene mixing over a salinity range of 5 to 35‰. Resting metabolic rate (RMR), as measured by oxygen uptake (V˙ _O2), was highest in the hybrids. Differences in V˙ _O2 could not be accounted for by differences in systemic adaptations (e.g. heart rate or scaphognathite frequency); therefore it was concluded that they were based on potential underlying differences in carbohydrate metabolism. The higher RMR in the hybrids was attributed to a higher metabolic cost of maintenance, resulting from the breakup of coadapted gene complexes as a result of introgressive hybridization. Received: 22 July 1996 / Accepted: 17 October 1996     

Physiological energetics of cultivated mussel (Mytilus edulis) populations in two Scottish west coast sea lochs

Physiological responses of suspended cultured mussels, Mytilus edulis L., in two Scottish sea lochs (Lochs Etive and Leven) were investigated in their native and transplanted environments, after 15 d, 4.5 mo and 1 yr acclimatization, during the main growing season of May to September 1992. These measurements were integrated by means of the balanced energy equation, and scope for growth was calculated to assess the performance of each stock. Transplanted mussels showed clear signs of stress during the first 15 d after transfer, with low clearance rates and energy retention and high rates of respiration and nitrogen excretion. There were significant differences in some of the physiological responses (clearance, respiration and excretion rates), scope for growth and growth efficiency between the native populations, with the responses of mussels in Loch Etive being more favourable than those in Loch Leven. With newly transplanted mussels after 15 d acclimatization, almost all these measurements also differed significantly from native mussels in their host site and the original stocks but, with the exception of ammonia excretion rates, all the variables of cross-transplanted mussels after 4.5 mo acclimatization were the same as those of the native stock at the host site. Good agreement between observed long-term growth rates and estimated scope for growth suggests that, like growth rate, differences in physiological responses are mainly controlled by environmental factors; i.e., stock or origin had no significant influence on variations in physiological response (except ammonia excretion). The results further indicate that scope for growth estimated during the main growing season can be used to assess the actual growth rate and to compare sites for on-growing.     

Feeding and predator-avoidance by the rose anemone Urticina piscivora

We investigated the influence of feeding rate on the behavior of the rose anemone Urticina piscivora. We predicted that anemones would travel less and be less willing to detach from the substrate in response to contact with a predator (Dermasterias imbricata) as feeding rate was experimentally increased. As predicted, anemones receiving 0 mussels (Mytilustrossulus) daily moved significantly more than those receiving either 1 or 2 mussels daily. Anemones receiving 0 mussels daily also detached significantly quicker in response to predator contact compared to the 1 mussel per day group, which in turn detached significantly quicker than did the 2 mussels per day group. A field test of the same prediction gave mixed results. We also examined whether anemones could detect feeding rates of neighbors. Movement rates of anemones receiving 1 mussel daily when neighbors received 0 mussels daily were compared with those when neighbors received 2 mussels daily. We predicted that if the subjects could detect feeding rates of neighbors, they should move less when neighbors received less food than themselves (since their position is relatively good) than when neighbors received more food. The results support this interpretation. Received: 21 August 1996 / Accepted: 22 October 1996     

Assessment of sperm competition by European bitterling,<Emphasis Type="Italic"> Rhodeus sericeus</Emphasis>

We investigated male assessment of sperm competition in the bitterling, Rhodeus sericeus, a freshwater fish that spawns on the gills of living unionid mussels. Field experiments showed males increased their inspection rate of mussels into which a testis solution containing sperm had been experimentally released. Males avoided leading females to mussels that contained high numbers of embryos, but did not alter their leading behavior in response to the presence of sperm. In laboratory experiments males also increased their inspection rate of mussels into which a testis solution had been released and also failed to alter their leading behavior in response to the presence of sperm in mussels. However, males avoided leading females to mussels in close proximity to other males, and thereby may avoid sperm competition. In a second field study, territorial males were shown to ejaculate into mussels at a low rate in the absence of competitors, increase the frequency of ejaculations in competition with a rival, then decrease relative ejaculate expenditure as the number of competing males increased. Observed data were shown to be significantly correlated with predicted estimates of ejaculate expenditure for a model of sperm competition intensity. We discuss our results in the context of adaptive responses of males to sperm competition.     

Lethal and sub-lethal effects of phototrophic endoliths attacking the shell of the intertidal mussel Perna perna

Animals that bore into calcareous material can cause considerable damage to molluscan shells. In contrast, smaller microbial phototrophic endoliths have until recently been thought of as relatively benign. Phototrophic endoliths (primarily cyanobacteria) infest the shells of 50 to 80% of midshore populations of the mussel Perna perna (L.) in South Africa. This infestation causes clearly visible shell degradation, and we record here ecologically important lethal and sub-lethal effects (e.g. changes in growth and reproductive output) of the endoliths on their mussel hosts. Endolith infestation reduced the strength of shells significantly and also affected shell growth. In situ marking of shells, using the fluorochrome calcein, showed that infested and non-infested mussels increased in shell length at the same rate. However, the rate of increase in shell thickness (associated with shell repair) was significantly faster in infested than in uninfested individuals. This increase in the rate of shell thickening was not sufficient to compensate for rapid endolith-induced shell degradation and, around the site of adductor muscle attachment, infested shells were thinner than their uninfested counterparts. The shells of 18% of recently dead mussels had holes induced by endolith erosion. This effect was highly size dependent, and the proportion of mortality due to endoliths rose to almost 50% for the largest mussels. The re-routing of energy due to shell repair had important sub-lethal effects on the reproductive rates of mussels. During the reproductive period, mean dried flesh mass for large (>70 mm), non-infested P. perna was substantially higher than for infested individuals. This difference was almost entirely due to differences in gonad mass, which was approximately 100% higher for non-infested mussels. We conclude that, by attacking the shell, phototrophic endoliths reduce both the longevity and reproductive output of large mussels on the midshore. Received: 26 January 1999 / Accepted: 17 August 1999     

Uptake and depuration of cesium in the green mussel Perna viridis

 The accumulation and depuration of Cs in the green mussels (Perna viridis) commonly found in the subtropical and tropical waters were studied under the laboratory conditions using radiotracer techniques. Following an initial rapid sorption onto the mussel's tissues, uptake of Cs exhibited linear patterns over a short exposure time (8 h) at different ambient Cs concentrations. The concentration factor was independent of ambient Cs concentration. The calculated uptake rate and initial sorption constant of Cs were directly proportional to the ambient Cs concentration. The calculated uptake rate constant from the dissolved phase in the mussels was as low as 0.026 l g⁻¹ d⁻¹. Uptake rates of Cs in the mussels were inversely related to the ambient salinity. Uptake increased about twofold when the salinity was reduced from 33 to 15 ppt. The effect of salinity on Cs uptake was primarily due to the change in ambient K⁺ concentration. The uptake rate decreased in a power function with increasing tissue dry weight of the mussels, although the initial sorption was not related to the mussel's body size. The efflux rate constant of Cs in the mussels was 0.15 to 0.18 d⁻¹, and was the highest recorded to date among different metals in marine bivalves. The efflux rate constant also decreased in a power function with increasing tissue dry weight of mussels. A simple kinetic model predicted that the bioconcentration factor of Cs in the green mussels was 145, which was higher than measurements taken in their temperate counterparts. The bioconcentration factor also decreased in a power function with increasing tissue dry weight of mussels. Received: 27 October 1999 / Accepted: 16 June 2000     

Use of Fatty Acid Analysis to Determine Dispersal of Caspian Terns in the Columbia River Basin,U.S.A.

Abstract: Lethal control, which has been used to reduce local abundances of animals in conflict with humans or with endangered species, may not achieve management goals if animal movement is not considered. In populations with emigration and immigration, lethal control may induce compensatory immigration, if the source of attraction remains unchanged. Within the Columbia River Basin (Washington, U.S.A.), avian predators forage at dams because dams tend to reduce rates of emigration of juvenile salmonids (Oncorhynchus spp.), artificially concentrating these prey. We used differences in fatty acid profiles between Caspian Terns (Hydroprogne caspia) at coastal and inland breeding colonies and terns culled by a lethal control program at a mid‐Columbia River dam to infer dispersal patterns. We modeled the rate of loss of fatty acid biomarkers, which are fatty acids that can be traced to a single prey species or groups of species, to infer whether and when terns foraging at dams had emigrated from the coast. Nonmetric multidimensional scaling showed that coastal terns had high levels of C₂₀ and C₂₂ monounsaturated fatty acids, whereas fatty acids of inland breeders were high in C18:3n3, C20:4n6, and C22:5n3. Models of the rate of loss of fatty acid showed that approximately 60% of the terns collected at Rock Island Dam were unlikely to have bred successfully at local (inland) sites, suggesting that terns foraging at dams come from an extensive area. Fatty acid biomarkers may provide accurate information about patterns of dispersal in animal populations and may be extremely valuable in cases where populations differ demonstrably in prey base.     

An integrative response by Mytilus chilensis to the toxic dinoflagellate Alexandrium catenella

Physiological responses of Mytilus chilensis exposed to the toxic dinoflagellate Alexandrium catenella were measured over 21 days in the laboratory and were compared with control mussels not exposed to the dinoflagellate. Mussels were collected from culturing ropes at Yaldad Bay, southern Chile (43o08′S 73o44′W), in August 2004 and acclimated to laboratory conditions for one week prior to the experiment. After 8 days, the paralytic shellfish poisoning (PSP) toxins (i.e. saxitoxin) in the tissues of exposed mussels exceeded safe levels for human consumption. Clearance rates, ingestion of organic matter, and absorption efficiency of exposed mussels were significantly lower than those of controls on day 0, but this was followed by an increase on day 3. The exposed mussels also increased their excretion rate over time, and this increase was significantly correlated with the accumulation of PSP toxins in their tissues. Oxygen consumption was not affected by the PSP toxins. The scope for growth (SFG) on day 0 was negative in exposed mussels, but it increased during the experiment. Although feeding activity and absorption efficiency were adversely affected during the first few days of exposure to PSP toxins from A. catenella in the laboratory, the M. chilensis cultured in Yaldad Bay may have evolved mechanisms that allow them to exploit the toxic dinoflagellate as a food source.     

Rapid assessment of metabolic activity in marine microalgae: application in ecotoxicological tests and evaluation of water quality

A new method for the assessment of the effects of several contaminants on marine microalgae,Tetraselmis suecica (Kyling) Butcher,Skeletonema costatum (Grev.) Cleve, andProrocentrum lima (Ehrenberg) Dodge was developed in 1990. The method is based on the measurement of cell esterase activity using a fluorimetric stain, fluorescein diacetate (FDA), selected from amongst three stains (FDA, Neutral Red, thiazolyl tetrazolium bromure) for its higher sensitivity. Biochemical (K _m,V _max) and physiological (growth, specific activity) aspects of the enzymatic activity as revealed by the FDA method are discussed. Different categories of compounds (weed-killers, insecticides, metals) and some water samples from Seine Bay were tested for their toxic effects on microalgae. Experiments were performed on microplates using a fluorimetric microplate reader. The various steps of the experiments and data-processing were controlled by software. Applications of the system to rapid ecotoxicological tests (determination of the IC₅₀, i.e., the concentration at which 50% inhibition of growth occurs) and to the assessment of environment quality by studying the toxic effect of water samples on microalgae are proposed.     

Biochemical biomarkers and overall health status of mussels Mytilus galloprovincialis exposed to nickel and chromium

This study examined the effects of nickel and chromium exposure on two biochemical biomarkers, glutathione peroxidase (GPX) and acetylcholinesterase (ACHE), in parallel with an indicator of overall health, Scope for Growth (SFG), in the mussel Mytilus galloprovincialis to evaluate their ecological relevance and to assess their potential use as biomarkers of Ni and Cr pollution. SFG integrates major physiological responses into an index that represents the energy status of the animals and is considered an ecologically relevant biomarker. Following exposure to a range of Ni and Cr concentrations effects of Ni and Cr were evident on GPX and SFG but not on ACHE. Accumulation in the mussels’ tissues increased according to exposure concentrations. Adverse effects on the energy status and induction of GPX were found at the higher exposure concentrations while possible hormetic effects on the energy status and moderate GPX suppression were associated with lower exposure concentrations. Nevertheless GPX was negatively correlated with SFG suggesting that GPX responses reflect the overall health of the organisms and thus show potential as an early warning signal of population level effects of Ni and Cr. On the other hand, our results indicate that ACHE is not a suitable biomarker for Ni and Cr pollution.     

Investigation of the sources of aliphatic hydrocarbons in the mussel Mytilus edulis from north sea oil production platforms by Capillary glc and CGCMS†

Aliphatic hydrocarbons isolated from mussels collected over a 20 month period from three North Sea Forties field oil production platforms have been examined by capillary gas chromatography and gas chromatography‐mass spectrometry.

The biological hydrocarbons consist predominantly (300–700 μh g^‐1 lipid) of C₃₁ and C₃₃ n‐alkenes with 2, 3 and 4 double bonds probably derived from a dietary intake of Emiliania huxleyi, a microscopic coccolithophorid alga. In some of the mussels C₁₈, C₂₀ and C₂₂ n‐alkanes are present in unusually high abundance compared to their odd carbon number homologues. Possible reasons for this distribution are discussed.

The concentrations of fossil fuel hydrocarbons (mainly unresolved complex mixtures) in the mussels ranged from 330 to 5,298 μg g^‐1 of lipid. No significant increase in values was detected two months after the start of discharge of treated Forties production water from the Forties D platform.

To determine the pollutant hydrocarbon sources, the sterane and terpane distributions of hydrocarbon fractions isolated from the mussels and from a number of fossil fuels, including Forties crude oil, were examined by mass fragmentography. The steranes in the mussels from the Forties C and D platforms contained higher proportions of regular 14α(H), 17α(H) components than Forties oil in which the steranes were mainly 13ß(H), 17α(H) diasteranes. In addition, the terpane distributions of the mussel fractions differed markedly from that of Forties crude oil, particularly in the relative abundance of diterpanes to triterpanes, which was higher in the mussel fractions than in the oil, and in the absence of 17α(H), 18α(H), 21ß(H)‐18, 30‐bisnorhopane, a known component of Forties crude. Furthermore, the ratio of C₂₉ to C₃₀ triterpanes was greater than unity in the mussels but much less than unity in the Forties oil. These data preclude Forties or other similar North Sea crudes as the major source of pollutant aliphatic hydrocarbons in the mussels. The relative concentrations and distributions of steranes and triterpanes suggest that the most likely source of pollutants is a Middle Eastern based oil derived either from rig activities, or from background pollution in the North Sea. The low concentration of ≥ C₂₇ steranes in the more polluted mussels (e.g. Forties B; UCM > 200 ppm dry wt.) suggests that gas oils used on the platforms may be the major source of petrogenic hydrocarbons in these samples.     

The importance of methane and thiosulfate in the metabolism of the bacterial symbionts of two deep-sea mussels

Undescribed hydrocarbon-seep mussels were collected from the Louisiana Slope, Gulf of Mexico, during March 1986, and the ultrastructure of their gills was examined and compared to Bathymodiolus thermophilus, a mussel collected from the deep-sea hydrothermal vents on the Galápagos Rift in March 1985. These closely related mytilids both contain abundant symbiotic bacteria in their gills. However, the bacteria from the two species are distinctly different in both morphology and biochemistry, and are housed differently within the gills of the two mussels. The symbionts from the seep mussel are larger than the symbionts from B. thermophilus and, unlike the latter, contain stacked intracytoplasmic membranes. In the seep mussel three or fewer symbionts appear to be contained in each host-cell vacuole, while in B. thermophilus there are often more than twenty bacteria visible in a single section through a vacuole. The methanotrophic nature of the seep-mussel symbionts was confirmed in ¹⁴C-methane uptake experiments by the appearance of label in both CO₂ and acid-stable, non-volatile, organic compounds after a 3 h incubation of isolated gill tissue. Furthermore, methane consumption was correlated with methanol dehydrogenase activity in isolated gill tissue. Activity of ribulose-1,5-biphosphate (RuBP) carboxylase and ¹⁴CO₂ assimilation studies indicate the presence of either a second type of symbiont or contaminating bacteria on the gills of freshly captured seep mussels. A reevaluation of the nutrition of the symbionts in B. thermophilus indicates that while the major symbiont is not a methanotroph, its status as a sulfur-oxidizing chemoautotroph, as has been suggested previously, is far from proven.