镉和铅对菲律宾蛤仔脂质过氧化及抗氧化酶活性的影响

为了研究亚致死浓度的重金属对海洋贝类的毒性效应,探讨其可能的作用机理,在实验生态条件下以菲律宾蛤仔(Ru-ditapes philip pinarum)为目标生物,采用半静态毒性实验方法,研究了不同浓度Cd2+(0.0948、0.237和0.474mg·L-1)和Pb2+(0.276、0.690和1.380mg·L-1)对菲律宾蛤仔鳃和消化腺组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性以及丙二醛(MDA)含量的影响。结果显示:(1)鳃和消化腺中的抗氧化酶及MDA的变化呈现出类似的趋势,在胁迫初期,各浓度处理组菲律宾蛤仔鳃和消化腺组织SOD和CAT与对照组相比活性都显著升高(P<0.01),呈现出明显的诱导效应,而MDA含量与对照组相比差异不显著(P>0.05);随着胁迫时间的延长,高浓度Cd2+(0.474mg·L-1)和Pb2+(1.380mg·L-1)处理组中SOD、CAT活性快速下降,与对照组相比差异显著;低浓度处理组中的抗氧化活性虽然也较对照组有所下降,但总体下降幅度不如高浓度组明显,并且所历暴露时间较长。各浓度处理组中MDA含量变化一致,均呈现出先升高后降低的趋势,且含量均高于对照组。(2)通过相关性分析,菲律宾蛤仔消化腺组织中的SOD活性与Cd2+浓度的相关性大于鳃组织,与Pb2+浓度的相关性则小于鳃组织;消化腺组织中的CAT活性与Cd2+、Pb2+浓度呈抛物线型相关,与鳃组织CAT活性相关性不十分显著。这说明消化腺组织中SOD活性对Cd2+的敏感性大于鳃组织,消化腺组织中CAT活性对Cd2+、Pb2+的敏感性大于鳃组织。因此,菲律宾蛤仔消化腺中SOD、CAT对水环境中的重金属反应敏感,且存在一定的剂量-效应关系,消化腺组织中SOD和CAT活性与其他敏感性指标一起可以作为指示早期海洋重金属污染的生物标志物。     

Uptake and incorporation of mercury into mercury-binding proteins of gills of Mytilus edulis as a function of time

Individuals of Mytilus edulis (collected from Sequim Bay, Washington, in April and August, 1980) were exposed to 5 g l^-1 mercury as HgCl₂ for 28 d. Gill mercury accumulation, mercury incorporation into the soluble fraction and low molecular weight, mercury-binding proteins of gills, and induction of these mercury-binding proteins were determined as a function of time. Short-term mercury uptake rates of excised gills were also determined for mussels sampled at intervals during exposure. Gill mercury accumulation occurred in three phases, represented by net uptake phases initially (up to Day 4) and toward the end of the exposure (Days 15 to 28), and an intermediate stable phase (Days 4 to 15). The stable phase was associated with induction of the predominant mercury-binding proteins and mercury incorporation into the proteins. After Day 15, the mercury-binding proteins were saturated and spillover of mercury into high molecular weight proteins had occurred. This was associated with saturation of the soluble fraction, increases of mercury on particulate fractions, and a loss of the ability of gills to maintain stable mercury concentrations. Mercury uptake rates of gills were not affected by the 28 d exposure of the whole organism.     

Effect of copper on ion- and osmoregulation in the shore crab Carcinus maenas

The uptake and effect of dissolved copper on regulation of hemolymph osmolality and Na⁺, K⁺, Cl^-, Ca⁺⁺, and Mg⁺⁺ concentrations in the shore crab Carcinus maenas (L.) were determined at 400 mOsm (=14 S) ambient salinity. One mg Cu l^-1 resulted in 50% mortality in 11 to 22 d; the highest sensitivity was observed around the moulting period. 0.25 and 0.5 mg Cu l^-1 were not lethal after a onemonth exposure. Ten, 1, and 0.5 mg Cu l^-1 altered regulation of osmolality, Na⁺, K⁺, and Cl^- concentrations, while 0.25 mg Cu l^-1 did not. Exposure to 1 mg Cu l^-1 reduced hemolymph osmolality and Na⁺, K⁺, and Cl^- concentrations to 80 to 90% of controls within 4 to 6 d and no further reduction was observed during a 21-d exposure. The effect of various copper concentrations on these four parameters were almost identical and the highest sensitivity was observed around the moulting period. Hemolymph calcium levels increased 20 to 80% in crabs exposed to 1 mg Cu l^-1, while they decreased 20 to 30% in crabs exposed to 0.5 mg Cu l^-1. Prolonged exposure to copper caused 20 to 70% reductions in hemolymph magnesium levels. Crabs exposed to 0.5 mg Cu l^-1 for 29 d accumulated copper in hepatopancreas, gills, carapace, heart, testes, and hypodermis, but not in muscles and hemolymph. Increased copper levels in crabs exposed to 0.25 mg Cu l^-1 were observed in hepatopancreas, gills, and carapace only. The present results suggest that effects of copper on ion regulatory processes in part explain why the toxicity of copper towards euryhaline invertebrates increases at low salinities.     

Inhibition of Na+/K+-ATPase and of active ion-transport functions in the gills of the shore crab Carcinus maenas induced by cadmium

Inhibition of Na⁺/K⁺-ATPase from gill plasma membranes of the shore crab Carcinus maenas by cadmium was investigated and compared with inhibitory effects by known antagonists (ouabain and Ca²⁺). For comparative considerations the Cd²⁺-inhibition of the enzyme from dog kidney was also tested. Na⁺/K⁺-ATPase from dog kidney and from crab gill differed greatly in sensitivity against ouabain. The inhibition constant K _i of the dog enzyme amounted to 9.1 × 10⁻⁷ mol l⁻¹, i.e. more than 300-fold smaller than the K _i of 2.9 × 10⁻⁴ mol l⁻¹ determined for the crab enzyme. Ca²⁺ inhibited the activity of Na⁺/K⁺-ATPase from crab gill plasma membranes with a K _i of 4.3 × 10⁻⁴ mol l⁻¹. The Na⁺/K⁺-ATPase from crab gill was inhibited by Cd²⁺ with a K _i of 9.1 × 10⁻⁵ mol l⁻¹. Cd²⁺ inhibited the Na⁺/K⁺-ATPase from dog kidney with a K _i (6.4 × 10⁻⁵ mol l⁻¹) comparable to that observed in the crab gill enzyme. Under experimental conditions Cd²⁺-inhibition of Na⁺/K⁺-ATPase was irreversible. Repeated washing, centrifugation and homogenization of the plasma membranes (four times) with Cd²⁺-free buffer did not restore any activity lost in the presence of 1 × 10⁻³ mol l⁻¹ Cd²⁺. Since ouabain-insensitive (nonspecific) ATPases in the plasma membrane fraction of crab gills were inhibited by Cd²⁺ in the same way as Na⁺/K⁺-ATPase, the heavy metal is considered as an unspecific ATPase inhibitor. Comparing these results with literature data on Cd²⁺-binding to electrophoretically separated proteins suggests that Na⁺/K⁺-ATPase is a Cd²⁺-binding enzyme. The results obtained on Na⁺/K⁺-ATPase were reflected by Cd²⁺-inhibition of the branchial ion-transport functions depending on this enzyme. The transepithelial short-circuit current of isolated gill half lamellae, a direct measure of area-specific active ion uptake, and the transepithelial potential difference of isolated, perfused whole gills, also indicative of active ion uptake, were inhibited by the heavy metal in a time- and dose-dependent mode. Remarkably these inhibitions were also irreversible. These findings are ecologically and biomedically significant: even when the actual environmental or tissue concentrations measured are low, biological microstructures such as Na⁺/K⁺-ATPase may accumulate the heavy metal by tight binding over prolonged periods until the first inhibitory effects occur. Received: 25 June 1997 / Accepted: 25 August 1997     

Effects of lead exposure on redox status,DNA and histological structures in Venus verrucosa gills and digestive gland

ABSTRACT

Lead (Pb) is one of the most toxic heavy metals that affect the physiological status of aquatic organisms. The present investigation evaluated the possible toxic effect of lead chloride (PbCl₂) on biomarkers responses, DNA damage and histological alterations in Venus verrucosa gills and digestive gland. Three concentrations of PbCl₂ (D1:1µgL^?1, D2: 10µgL^?1 and D3: 100µgL^?1) were chosen for V. verrucosa exposure during six days. At the end of the trial, it was found that Pb tended to accumulate in both gills and digestive gland in a dose-dependent manner. However, gill tissues exhibited the highest metal burden. Our results showed an increase of malondialdehyde, protein carbonyls and advanced oxidation protein product levels in both organs following PbCl₂exposure. The induction of both non-enzymatic and enzymatic antioxidant systems; as well as the decrease of the acetylcholinesterase activity and degradation of DNA structure was recorded in the gills and digestive gland. The histopathological alterations observed in gills (disruption of lamellas and cilia filaments?…) and digestive gland (lumens occlusion, necrosis and fibrosis) confirmed the aforementioned results. Our data highlighted the short-term toxicity effects of PbCl₂ on V. verrucosa and pointed out a high sensitivity of gills towards this metal.     

Sodium regulation in the shore crab Carcinus maenas as related to ambient salinity

The activity of Na–K-ATPase was measured in crude homogenates prepared from various organs (leg muscle, pincer muscle, heart, testes, digestive gland, hypodermis, gills 1–9) of shore crabs, Carcinus maenas L., acclimated to salinities ranging between 10 and 50 S (in steps of 10 S). In all salinities tested, Na–K-ATPase activity was highest in posterior gills 7–9 (10–12 mol P_i mg protein^-1 h^-1), followed by anterior gills 1–6 (ca. 2.5 mol P_i mg protein^-1 h^-1) and the other organs (in most cases far below 2mol P_i mg protein^-1 h^-1). In gills only, Na–K-ATPase activity was salinity-dependent, with the highest values in the lowest salinities and vice versa. In gills 7–9, Na–K-ATPase activity was increased more than threefold following a reduction in salinity from 50 to 10 S. Na–K-ATPase activity, expressed as percentage of total ATPase activity, amounted to 60–80% in gills, about 60% in hypodermis and 20–40% in the other organs. Ouabain, a specific inhibitor of Na–K-ATPase activity, reduced serum osmolalities in crabs kept at 9–10 S only when injected into the hemolymph (1 and 5 · 10^-5 M), but had no effect when dissolved in ambient water (10^-4 M). The results obtained underline that crustacean gills are the main organs for ionic regulation, and confirm the hypothesis of the central role of the Na–K-ATPase in active Na uptake as the basic mechanism of hyperregulation in dilute media. Reduction of serum osmolalities following injection of ouabain into the hemolymph confirms previous reports on localization of the sodium pump in the basolateral parts of epithelial cells.     

三甲基氯化锡对斑马鱼(Danio rerio)生理生化特性的影响

为初步探讨三甲基氯化锡(trimethyltin chloride,TMT)对鱼类的毒性效应以及评价环境中TMT的安全性,采用静态鱼类急性毒性试验法测定了TMT对斑马鱼(Danio rerio)生理生化指标的影响;参考TMT的96 h-LC50值,设定3个浓度(0.39、0.78和1.17 mg·L-1)处理斑马鱼,测...     

Transbranchial potentials and ion fluxes across isolated,perfused gills of Uca rapax

Transbranchial potentials (TP) and sodium or chloride fluxes were measured in an apparatus designed for the simultaneous perfusion of eight isolated gills of Uca rapax. In anterior gills perfused with U. rapax–saline (US) the TP varied almost linearly from-7.5 to +10 mV inside, and in posterior gills from +2 to-8.5 mV (inside), on exposure to salinities ranging from 8.7 through 52, i.e. 25 to 150% seawater (100%=34.6 S). Sodium influx and efflux in anterior gills exposed to US, 8.7 or 43.3 S (0.7 to 4.0 mmol h^–1 g^–1 dry wt) were always greater than in posterior gills (0.5 mmol h^–1). The chloride fluxes were slightly smaller than sodium fluxes in anterior gills, while in the posterior gills the chloride influx (2.8 to 4.6 mmol h^–1) was always larger than chloride efflux (0.6 to 1.1 mmol h^–1) or the sodium fluxes. At least three ion-transport mechanisms may be present in these gills: (1) an internal ( = basolateral), ouabain-sensitive Na⁺, K⁺ pump, restricted to anterior gills; (2) a furosemide-sensitive Na⁺, K⁺, 2Cl^– (plus water) transporter, apparently restricted to posterior gills, and (3) a Na⁺ exchanger (and possibly other as yet unidentified ion transporters, as suggested by large increases of the chloride influxes caused by amiloride), probably located on the apical membranes of the epithelial cells of both gill types. The differential selectivity of the gills of U. rapax for sodium or chloride may limit the transbranchial movements of either ion, without a reduction of the overall permeability of these crabs.Communicated by N.H. Marcus, Tallahassee     

Quantitative determination of metallothionein-like proteins in mussels. Methodological approach and field evaluation

Electrochemical quantitation of metallothionin-like proteins (MLP) in mussels was based on the determination of their constituent cysteinyl residues according to Brdika's catalytic reaction. Calibration was performed by an internal MLP standard isolated from the digestive gland of Mytilus galloprovincialis for which protein concentration had been estimated by Bradford's spectrophotometric method. For that purpose three metal-binding proteins [MLP-I, MLP-II and Cu-BP (binding protein)] were separated by DEAE-Sephadex A-25 chromatography from the digestive gland of mussels previously exposed to Cd. The most negatively changed MLP-II fraction was characterized by the fact that it contained the largest amount of both total metal and sulphydryl (-SH) content per mass of protein, although this was approximately two times lower than the-SH level of commercially available MT from rabbit liver. Exposure of mussels to a relatively low level of cadmium (0.2 g Cd l^-1) added into the seawater either by itself or as a mixture with other metals (2 g Cu l^-1 and 1.6 g Pb l^-1) resulted in a measurable level of MLP induction within 14 d in comparison to the control specimens. The effect of the metal mixture on MLP synthesis appears to be less than additive, suggesting a competitive interaction between metal ions for uptake and binding sites as well as differing potentials for MLP induction. Variations in the MLP content observed in the digestive gland of mussels seasonally collected from the same location are in the range 2.1±0.4 mg g^-1 on a wet weight basis. The methodological and conceptual aspects of the application of MLP induction in the Mytilus sp. as a biomarker in seawater trace metal monitoring are critically evaluated.     

Branchial and intestinal osmoregulatory acclimation in the four-eyed sleeper, <Emphasis Type="Italic">Bostrychus sinensis</Emphasis> (Lacepède), exposed to seawater

Bostrychus sinensis is a facultative air breather that inhabits waters of a wide range of salinities. This study aimed to elucidate whether branchial and intestinal osmoregulatory acclimation occurred in B. sinensis transferred from 5‰ water through a progressive increase in salinities to seawater. Our results indicate that B. sinensis acted as a hyperosmotic regulator in 5‰ water, but exhibited hypoosmotic hypoionic regulation in seawater. During short- (1 day) and medium- (10 days) term acclimation to seawater, there were only minor perturbations in plasma osmolality and [Na⁺], which returned to control levels after 45 days of exposure to seawater. Branchial Na⁺/K⁺-ATPase activity was unaffected by 1, 10 or 45 days of exposure to seawater. However, prolonged (45 days) acclimation to seawater led to a significant increase in Na⁺/K⁺-ATPase α-subunit protein abundance. Taken together, these results indicate that there could be changes in the expression of Na⁺/K⁺-ATPase isoforms and/or post-translational modification of Na⁺/K⁺-ATPase in the gills of fish exposed to seawater. Immunofluorescence microscopy revealed that acclimation to seawater for 10 days only resulted in no change in branchial Na⁺/K⁺-ATPase protein expression, but there were increases in protein expression of cystic fibrosis transmembrane regulator (CFTR)-like chloride channel and Na⁺:K⁺:2Cl⁻ cotransporter (NKCC; probably NKCC1). Indeed, NKCC was undetectable in gills of fish kept in 5‰ water by Western blotting, but it became weakly detectable in fish exposed to seawater for 10 days and prominently expressed in fish exposed to seawater for 45 days. Therefore, our results indicate that branchial CFTR-like chloride channel and NKCC1 were the determining factors in the transition between hyperosmotic regulation and hypoosmotic hypoionic regulation in B. sinensis. Furthermore, the intestine of B. sinensis also served as an important osmoregulatory organ, since there were significant increases in both the activity and protein abundance of intestinal Na⁺/K⁺-ATPase in fish acclimated to seawater for 45 days. The effectiveness of branchial and intestinal osmoregulatory acclimation in B. sinensis during seawater acclimation led to only a minor increase in plasma osmolality, and thus resulted in relatively unchanged free amino acid contents in muscle and liver.     

Community metabolism of adenylates by microheterotrophs from the Los Angeles Harbor and Southern California coastal waters

The natural concentration (S _n) of dissolved total adenylates TA(=AMP+ADP+ATP) in coastal seawater from a depth of 1 m at 5 stations (California, USA) sampled periodically for 1 yr had a mean value ±1 SD of 2.8±1.7 nmol TA1^-1. The specific uptake rates of TA by microheterotrophs at a station inside the Los Angeles Harbor and at a station 1.5 km offshore in the San Pedro Channel were studied by simple uptake and saturation-type kinetic analysis using ³H-AMP as a tracer. Within the harbor, the specific uptake rate (nmol TA 10⁹ cell^-1 h^-1) at S _n ranged 10-fold from 0.028 in December to 0.28 in August. K _t (half-saturation constant) values always exceeded theS _n concentrations in any given month, and were greater in the harbor than in the channel. Generally, over 80% of biological uptake of ³H-AMP was associated with organisms <1.0 m, a size class accounting for about 20% of the total particulate adenylate concentration in the 0.2 to 203 m size fraction. Assuming steady-state conditions for the dissolved adenylate pool, we propose a model in which losses from this pool are balanced by inputs to the pool through inefficient feeding, lysis and decomposition of particulate adenylates.     

Laboratory studies on the accumulation of inorganic mercury and methylmercury by the mussel (Mytilus galloprovincialis Lam) in relation to the presence of sediment

A comparative study on the accumulation of inorganic mercury and methylmercury by the mussel Mytilus galloprovincialis was based on nine days of exposure to 25 µg L^?1 HgCl₂ or to 2.5 µg L^?1 methylmercury in laboratory microcosms. Mercury (Hg) content was evaluated in the gills, digestive gland, and mantle. A higher accumulation occurred in the gills than in other tissues. The effect of the sediment on the bioaccumulation of the Hg species was evaluated. The results showed that the sediment accumulated the inorganic Hg more efficiently than the methylmercury. In both cases, the bioaccumulation in the tissues was reduced. The observed differences emphasized the need for caution when field results are compared with the results of laboratory experiments. The fate of either inorganic or organic Hg was depicted considering the absolute total amount given and the amount found in all the matrices (organism, sediment, and seawater).     

Protein kinase C and ion transport in the posterior gills of the Chinese crabEriocheir sinensis

The possible involvement of protein kinase C in control of ion transport was investigated on a preparation of isolated, perfused posterior gills of the Chinese crabsEriocheir sinensis (collected in 1989 from lakes near Emden, northern Germany) acclimated to fresh water. 1-oleyl-2-acetyl-sn-glycerol (OAG) and phorbol 12-myristate 13-acetate (PMA), two activators of protein kinase C, when added to the perfusion saline, induced depolarisation of the transepithelial potential difference (PD) and an increase in transepithelial Na⁺ influx. The observed increase was proportional to OAG concentration up to 250µM, with a 2.5× accelerated Na⁺ influx. OAG and PMA remained without effect on Cl^– fluxes. The observed effects were in agreement with an activation, via protein kinase C, of the Na⁺/K⁺ ATPase located on the serosal side of the epithelium.     

Principal Biogeochemical Factors Affecting the Speciation And Transport of Mercury through the terrestrial environment

It is increasingly becoming known that mercury transport and speciation in the terrestrial environment play major roles in methyl-mercury bioaccumulation potential in surface water. This review discusses the principal biogeochemical reactions affecting the transport and speciation of mercury in the terrestrial watershed. The issues presented are mercury-ligand formation, mercury adsorption/desorption, and elemental mercury reduction and volatilization. In terrestrial environments, OH^–, Cl^– and S^– ions have the largest influence on ligand formation. Under oxidized surface soil conditions Hg(OH)₂, HgCl₂, HgOH⁺, HgS, and Hg⁰ are the predominant inorganic mercury forms. In reduced environments, common mercury forms are HgSH⁺, HgOHSH, and HgClSH. Many of these mercury forms are further bound to organic and inorganic ligands. Mercury adsorption to mineral and organic surfaces is mainly dictated by two factors: pH and dissolved ions. An increase in Cl^– concentration and a decrease in pH can, together or separately, decrease mercury adsorption. Clay and organic soils have the highest capability of adsorbing mercury. Important parameters that increase abiotic inorganic mercury reduction are availability of electron donors, low redox potential, and sunlight intensity. Primary factors that increase volatilization are soil permeability and temperature. A decrease in mercury adsorption and an increase in soil moisture will also increase volatilization. The effect of climate on biogeochemical reactions in the terrestrial watershed indicates mercury speciation and transport to receiving water will vary on a regional basis.     

Conductive sodium entry in gill cells of the shore crab,Carcinus maenas

Isolated posterior gills of shore crabs, Carcinus maenas, collected from the Baltic Sea, were perfused and bathed with sea water and solutions of alkali chlorides. The preparation was used to measure fluxes of sodium from the external medium across the gills into the hemolymph and to determine transepithelial potential differences (PDs). Internally negative active transport PDs resulted from perfusion and bathing the gills symmetrically with the same medium (50% sea water). Passive (asymmetry) PDs following employment of 100% sea water as internal and 50% sea water as external medium were — in contrast to symmetry PDs — insensitive to cyanide and ouabain. This result indicates that the gill recognized the desired hyperosmotic state and responded by switching off the active transport component observed under symmetry conditions. Diffusional potential differences and fluxes of Na⁺ were inhibited by the externally applied diuretic amiloride. Gradients of pure alkali chlorides between medium and blood were accompanied by cation specific PDs. These PDs and their amiloride sensitivity were inversely related to the diameter of the unhydrated cation and allowed the calculation of the permeability sequence P: Li⁺>Na⁺>K⁺>Rb⁺>Cs⁺>Cl^-. The results obtained show that the permeability of the gills to cations greatly exceeds that to anions. In addition, these findings indicate that the initial amiloride-sensitive step in Na⁺ transport across the gill is not represented by an electroneutral sodium/proton exchange but by a conductive mechanism.     

Interactions of marine plankton with transuranic elements

In a series of laboratory experiments, the biokinetics of ²⁴¹Am, an important transuranium element, was studied in Meganyctiphanes norvegica, a euphausiid common in the northwestern Mediterranean. The euphausiids accumulated Am from water by passive adsorption onto exoskeletons, achieving wet weight concentration factors on the order of 10² after 1 wk exposure; concentration factors varied inversely with the size of the euphausiids and linearly with their surface area:wet weight ratios. Essentially all (96±10%) of the Am taken up from water was associated with the exoskeleton, so that negligible Am was retained by the euphausiids after molting. The retention half-time of Am in molts was 2.9 d. Euphausiids could also concentrate Am from feeding suspensions by ingesting Am-labelled diatom cells, although there was negligible Am assimilation (3±2% after 4 d feeding); after passage through the gut, virtually all (99%) of the ingested Am was defecated within 1 wk. The retention half-time of Am in fecal pellets was 41 and 51 d at 13° and 5°C, respectively. In oceanic waters, where the preponderance of ²⁴¹Am is in the dissolved phase, uptake of Am from water by euphausiids would be the dominant route of bioaccumulation. The results underscore the importance of sinking biogenic debris from zooplankters in mediating the vertical transport of Am in the sea. Given their retention half-times for ²⁴¹Am and their rapid sinking rates, fecal pellets and discarded molts have the potential to deliver most of their Am to the sediments.     

A kinetic and autoradiographic study of the direct assimilation of amino acids and glucose by organs of the musselMytilus edulis

The uptake of amino acids and glucose dissolved in sea water by different parts of common mussels (Mytilus edulis, L.) is studied from the first minutes up to 1 week. Autoradiography and donble countings on¹⁴C and²H labelled samples show that, in the first hour, the gills and mantle can concentrate several hundred times the dissolved nutritive molecules from very dilute solutions, whereas the digestive tract is not involved significantly during the first hours. An important -amylasic activity has been detected in the gills. The gill epithelium shows a strong positive reaction with mixtures used for the histochemical detection of chymotrypsin. This suggests that the digestion of small particles as well as the absorption of dissolved food might be initiated on the surface of the palleal-gill areas, and completed later in the hepatic caeca.     

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     

Possible causes of temporal fluctuations in primary production of the microphytobenthos in the Danish Wadden Sea

The effects of prior exposure to unlabelled naphthalene on the processes of uptake, tissue distribution and elimination of [1-¹⁴C] naphthalene by mussels (Mytilus edulis) were examined. Mussels collected from Whitsand Bay (S. W. England) in October 1980 were either held unexposed or exposed to unlabelled naphthalene (0.5 μg 1^?1) for 4 wk, prior to receiving a 4 h pulse of [1-¹⁴C] naphthalene in the medium (7 μg 1^?1). After 4 h exposure to [1-¹⁴C] naphthalene, the major tissues (digestive gland, gills, kidneys, mantle and remaining tissue) showed exponential depuration curves which could be resolved into two statistically significant components. The kinetics of elimination and the biological half-times of [1-¹⁴C] naphthalene in the tissues were determined. Mussels pre-exposed to unlabelled naphthalene had a significantly higher rate of elimination of [1-¹⁴C] naphthalene from the gills and kidneys. These results indicate that the toxicant, naphthalene, is actively excreted from the body by the gills and kidneys and the process is enhanced by prior exposure to the toxicant.     

Voltage-clamp measurements on single split lamellae of posterior gills of the shore crab Carcinus maenas

Single split lamella preparations of posterior gills of low-salt adapted shore crabs Carcinus maenas (collected from the Kiel Bay, Baltic Sea in 1991) were mounted in a micro Ussing-chamber. With NaCl salines on both sides we found an outside positive potential difference (PD) of 6.6±1.3 mV, a short-circuit current (I_sc) of-240±65 Acm^-2 and a resistance (R_t) of 25±3 cm² (n=8). Substitution of Cl^- (gluconate) on both sides of the preparation resulted in a decrease of I_sc by more than 90% at constant R_t. I_sc disappeared and R_t increased after substitution of Na⁺ (choline). When ouabain (2 mmol l^-1) was applied to the internal NaCl-saline, I_sc decreased and R_t remained unchanged. Internal addition of 0.1 mmol l^-1 acetazolamide left I_sc and R_t unaffected. Application of amiloride to the external NaCl saline resulted in a increase of both inward negative I_sc and R_t. The dose dependence of the diuretic showed a maximal effect between 50 and 200 mol l^-1 with a half-maximal blocker concentration (K_AMI) of ca. 10 mol l^-1. The results show that the split lamella preparation of posterior gills of C. maenas is a low resistance epithelium which is able to effect a massive, electrogenic and coupled absorption of Na⁺ and Cl^-.