Infestation of the surf clam Mesodesma donacium by the spionid polychaete Polydora bioccipitalis

The surf clam Mesodesma donacium is an economically important species for Chilean and Peruvian shellfisheries. This clam is often infested by Polydora bioccipitalis, a species belonging to the Spionidae, the most common parasitic polychaete group. To study this association, clams were sampled monthly over a one-year period in northern Chile. Collected clams covered the entire available size range and were classified into four infestation levels in order to study: (1) the relationship between prevalence of infestation (PI) and host size, (2) the temporal pattern of infestation events related to seasonal temperature changes, and (3) the relationship between infestation, body condition index (BCI) and gonado-somatic index (GSI). Additionally, growth rate and digging ability of clams with different infestation levels was studied. A logistic regression model best explained the relationship between PI and host size, with the smallest infested clam being 34 mm long and PI increasing steeply thereafter. Ontogenetic shifts in the habitat of the clam and ontogenetic changes, mainly in shell morphology, seem to explain the sigmoid pattern. Periods of increased shell blistering after infestation by P. bioccipitalis showed a similar seasonal pattern with GSI and BCI of non-infested clams, suggesting either an association between infestation ability and low condition of the clam or common environmental triggers for those factors. Heavily infested clams showed a significant lower BCI, growth rate and digging ability; however, given its low number, they are unlikely to be significant in terms of the local population survival. However, the infestation could play a key role in explaining mass mortality of northern populations during El Niño events, given the latitudinal differences in PI and the fact that infestation ability could be enhanced by increased temperature and facilitated in stressed clams.     

LPS胁迫下五个群体菲律宾蛤仔TYR基因在鳃和肝胰腺中表达特性研究

为了解TYR基因与蛤仔免疫的关系,本实验利用荧光定量PCR技术研究了菲律宾蛤仔五个群体(白蛤、白斑马蛤、斑马蛤、养殖和野生群体)的鳃组织和肝胰腺组织在LPS胁迫下TYR基因在不同时间(0 h、3 h、12 h、24 h、48 h)的表达特性。结果表明,在LPS注射后鳃组织中TYR6基因表达水平在白蛤和白斑马蛤3 h、6 h、12 h,野生蛤仔3 h,斑马蛤3 h、6 h,养殖群体6 h、12 h显著上调(P<0.05),3h野生蛤仔和斑马蛤达到峰值, 6 h白蛤、白斑马蛤、养殖群体达到峰值(P<0.05);在肝胰腺中,养殖群体和白斑马蛤6 h,白蛤6 h、24 h,野生群体24 h,斑马蛤3 h显著上调, 3 h野生蛤仔和斑马蛤达到峰值,6h养殖群体、白蛤、白斑马达到峰值(P<0.05);鳃组织中TYR10基因表达水平在白蛤、野生群体和白斑马蛤3 h,养殖群体3 h、6 h,斑马蛤3 h、12 h显著上调(P<0.05),肝胰腺组织中TYR10基因表达水平在白蛤3 h,野生群体3 h、6 h、12 h,斑马蛤、养殖群体6 h显著上调(P<0.05),推测T...     

Growth performance of the clam, Meretrix meretrix, breedingselection populations cultured in different conditions

A breeding-selection program for the clam,Meretrix meretrix,was conducted since 2004.Two of the selection populations were generated with the shell color pattern as an additional selection criterion and named as SP(purple stripes)population and SB(black dots)population.The third-generation SP and SB populations(08G3SP and 08G3SB,respectively)were cultured at two commercial clam farms and a nursery pond and their shell lengths were compared.08G3SB clams had significantly larger sizes than 08G3SP clams at commercial clam farms(p<0.05),yet 08G3SB individuals were significantly smaller than 08G3SP individuals at the nursery pond(p<0.05).Then,we examined the growth of the fourth-generation SP and SB populations(10G4SP and 10G4SB,respectively)at a commercial farm,and found that the shell lengths of the 10G4SB clams increased at a significantly higher growth rate than the 10G4SP clams(p<0.05)from May to September,when the water temperature was between 24.2–27.5 C,while 10G4SB lost the growth advantage in the other months.These results indicated that SP and SB populations responded differently to environmental factors,so it is beneficial for the clam farmers to select a suitable population according to the culture environment.Furthermore,a diallel mating of the SB and SP populations was designed to investigate whether their hybrid population would show heterosis.However,the heterosis was not shown in this study,which might result from the slight genetic divergence between SB and SP populations.     

Tapes philippinarum seed exposure to metals in polluted areas of the Venice lagoon

The concentration changes of 12 metals (As, Hg, Cd, Cr, Co, Cu, Pb, Mn, Ni, V, Zn, Fe, and Hg) in the soft tissues of Tapes philippinarum during growth were investigated. Clams were seeded in two contaminated areas of the Venice lagoon (San Giuliano and Fusina) and in an area of the Marano lagoon (Lignano Sabbiadoro) close to the clam-farm where the seed was produced. Metal trends were very different according to the considered element, the study site and the growth period.Arsenic was always higher in clam tissues than in surface sediments and suspended particles in all the three stations. Mercury, Cd and Cu were higher in the clams from Marano and Fusina but not in those from San Giuliano. Zinc and Co in clams exceeded the concentrations in the sediment and suspended particles only at Marano. The other elements (Cr, Pb, Mn, Ni, and Fe) were always higher in SPM and sediments. In general metal concentrations in clams were more highly correlated to concentrations in the suspended particles rather than in the surface sediments and in suspended clams rather than in bottom clams, nevertheless significant differences between stations and contaminants were found. Metal concentrations in clams were always lower than the European regulatory limits.     

A theoretical individual-based model of Brown Ring Disease in Manila clams,Venerupis philippinarum

An individual-based mathematical model was developed to investigate the biological and environmental interactions that influence the prevalence and intensity of Brown Ring Disease (BRD), a disease, caused by the bacterial pathogen, Vibrio tapetis, in the Manila clam (Venerupis (= Tapes, = Ruditapes) philippinarum). V. tapetis acts as an external microparasite, adhering at the surface of the mantle edge and its secretion, the periostracal lamina, causing the symptomatic brown deposit. Brown Ring Disease is atypical in that it leaves a shell scar that provides a unique tool for diagnosis of either live or dead clams. The model was formulated using laboratory and field measurements of BRD development in Manila clams, physiological responses of the clam to the pathogen, and the physiology of V. tapetis, as well as theoretical understanding of bacterial disease progression in marine shellfish. The simulation results obtained for an individual Manila clam were expanded to cohorts and populations using a probability distribution that prescribed a range of variability for parameters in a three dimensional framework; assimilation rate, clam hemocyte activity rate (the number of bacteria ingested per hemocyte per day), and clam calcification rate (a measure of the ability to recover by covering over the symptomatic brown ring deposit), which sensitivity studies indicated to be processes important in determining BRD prevalence and intensity. This approach allows concurrent simulation of individuals with a variety of different physiological capabilities (phenotypes) and hence by implication differing genotypic composition. Different combinations of the three variables provide robust estimates for the fate of individuals with particular characteristics in a population that consists of mixtures of all possible combinations.The BRD model was implemented using environmental observations from sites in Brittany, France, where Manila clams routinely exhibit BRD signs. The simulated annual cycle of BRD prevalence and intensity agrees with observed disease cycles in cultured clam populations from this region, with maximum disease prevalence and intensity occurring from December to April. Sensitivity analyses of modeled physiological processes showed that the level of hemocyte activity is the primary intrinsic determinant of recovery of infected clams. Simulations designed to investigate environmental effects on BRD suggested that the outcome of the host–parasite interaction is dependent on food supply (high values being favorable for the host) and temperature. Results of simulations illustrate the complex interaction of temperature effects on propagation and viability of the bacterium, on the phagocytic activity of the hemocytes, and on other physiological processes of the host clam. Simulations using 1 °C and 2 °C increases in temperature generally favored disease development, indicating that climate warming might favor the spread of BRD.     

缢蛏耐高盐性能及其与生长性状的相关性分析

缢蛏(Sinonovacula constricta)是我国重要的水产经济贝类,开展耐高盐新品种选育对平衡不同地区缢蛏良种养殖意义重大。本研究对不同家系缢蛏幼贝进行高盐胁迫,运用统计学方法和相关性分析探究了缢蛏耐高盐性能及其与生长性状的相关性。结果显示,缢蛏家系间壳长、壳高、壳宽和体质量表型值差异较大,体质量变异系数达15.53%。不同家系对高盐耐受性差异较大,家系间死亡率范围在0～100%。胁迫实验共持续9 d,各家系缢蛏首粒死亡时间在第1～9 d之间,而且死亡率较高的家系内缢蛏死亡时间离散程度较大,说明不同个体间高盐耐受性差异也较大。缢蛏开始出现大量死亡的时间主要在胁迫后第8 d,第9 d部分家系死亡率高于80%,说明缢蛏对高盐耐受存在一个阈值,超过该阈值会开始或大量死亡。皮尔逊和斯皮尔曼相关性分析显示,缢蛏存活时间和生长性状为正相关,其中存活时间与缢蛏壳长、体质量相关性均达到显著水平(P<0.05)。缢蛏存活率与体质量、生长速度均显著正相关(P<0.05)。整体而言,壳长较长和体质量较重的缢蛏个体耐高盐性能较强,在高盐环境中具有生长优势。本研究揭示了缢蛏的高盐耐受性及...     

Bioavailability and effects of sediment-bound TBT in deposit-feeding clams, Scrobicularia plana

Concentrations of tributyltin (TBT), dibutyltin (DBT), inorganic and total tin have been measured in water, sediments and deposit feeding clams, Scrobicularia plana, from 25 estuarine locations in England and Wales, in order to compare the bioavailability of different forms of the element. Abnormally high tin values in Scrobicularia from harbours and areas of high boating activity testify to the high bioaccumulation potential of organotins, notably TBT derived from anti-fouling paints, relative to inorganic tin.Infaunal deposit-feeding bivalves consistently contain higher concentrations of TBT than other benthic organisms indicating that sediments may be an important route for uptake. Significant correlations between sediment TBT values and residues in clams such as S. plana (r = 0·81, p < 0·001) support this hypothesis. Direct evidence of the bioaccumulation of sediment-bound TBT is provided from laboratory experiments in which Scrobicularia were exposed to TBT in water and in sediments, either separately or in combination. The results of these experiments clearly demonstrate a predominantly particulate component for TBT uptake in clams.The kinetics of TBT accumulation and elimination were studied in Scrobicularia exposed to sediments containing 0·3 and 1·0 μg/g TBT (as tin). Equilibrium concentrations in tissue are approached after 40 days.Sediments containing 10 μg/g TBT are acutely toxic to S. plana, although such concentrations are only likely to occur close to dockyards and large marinas. Preliminary laboratory and field observations suggest however, that clam populations could be affected at TBT concentrations in sediments of 0·3 μg/g and possibly lower.     

New polymorphic microsatellite markers derived from hemocyte cDNA library of Manila clam <Emphasis Type="Italic">Ruditapes philippinarum</Emphasis> challenged by the protozoan parasite <Emphasis Type="Italic">Perkinsus olseni</Emphasis>

Manila clam Ruditapes philippinarum is one of the most important benthic animals in the coastal north Pacific region, where clam populations have been mixed genetically through trade and aquaculture activities. Accordingly, identification of the genetically different clam populations has become one of the most important issues to manage interbreeding of the local and introduced clam populations. To identify genetically different populations of clam populations, we developed 11 expressed sequence tag (EST)-microsatellite loci (i.e., simple sequence repeat, SSR) from 1,128 clam hemocyte cDNA clones challenged by the protozoan parasite Perkinsus olseni. Genotype analysis using the markers developed in this study demonstrated that clams from a tidal flat on the west coast contained 6 to 19 alleles per locus, and a population from Jeju Island had 4 to 20 alleles per locus. The expected heterozygosity of the 2 clam populations ranged from 0.472 to 0.919 for clams from the west coast, and 0.494 to 0.919 for clams from Jeju Island, respectively. Among the 11 loci discovered in this study, 7 loci significantly deviated from the Hardy-Weinberg equilibrium after Bonferroni correction. The 5 loci developed in this study also successfully amplified the SSRs of R. variegatus, a clam species taxonomically very close to R. philippinarum, from Hong Kong and Jeju Island. We believe that the 11 novel polymorphic SSR developed in this study can be utilized successfully in Manila clam genetic diversity analysis, as well as in genetic discrimination of different clam populations.     

4种壳色蛤仔TYR基因的表达特性研究

为了解色素控制基因与壳色的关系,本实验用荧光定量聚合酶链式反应(Polymerase Chain Reaction,PCR)技术分析了酪氨酸酶基因(Tyrosinase,TYR)在菲律宾蛤仔(Ruditapes philippinarum)4种壳色蛤仔和7种组织中的表达特性。结果表明,酪氨酸酶基因在鳃、外套膜、闭壳肌、性腺、内脏团、水管和唇瓣中均有表达,其中外套膜中表达量较高,其次为鳃,在闭壳肌中表达量最少。不同的酪氨酸酶基因在4种壳色的表达量不同,酪氨酸酶基因TYR2、TYR3、TYR6和TYR9在白斑马蛤中表达量居高,且与斑马蛤和白蛤差异显著(P0.05)。TYR2、TYR6和TYR10在橙蛤中表达量最高,推测与橙色形成有关。TYR11在斑马蛤中表达量最高,且与白蛤和白斑马蛤差异显著(P0.05),推测与背景色形成有关。系统发育分析结果表明,TYR3与马氏珠母贝(Pinctada martensii)同源性最高为64%。TYR10与加州双斑蛸(Octopus bimaculoides)同源性最高,为53%。与长牡蛎进化关系最近。     

Biomarkers of biochemical and cellular stress in Carcinus maenas: an in situ field study

Biomarkers are useful diagnostic tools for identifying exposure to physical, chemical, and environmental stresses. In this study a suite of techniques—measurements of metallothionein, lysosomal integrity, and osmoregulatory ability—have been used in an attempt to provide an in situ assessment of environmental quality. Five estuarine sites were studied, two clean sites, two urban sites, and one intermediate. Shore crabs, Carcinus maenas, were collected and haemolymph taken for the lysosomal and osmoregulatory assays. Animals were killed and tissue preserved for metallothionein analysis. Lysosomal integrity was significantly different in crabs from clean sites and the urban sites (n = 40, p < 0.05). The potential value of the biomarker approach is that it could provide data for impact assessment and is applicable in the field.     

Spatial and temporal variations in the population structure of the soft-shell clam Mya arenaria in the Pomeranian Bay (southern Baltic Sea)

The population structure of Mya arenaria has been investigated in the sublittoral zone of the Pomeranian Bay (southern Baltic Sea). Box-corer samples were collected during a 1.5-year period in 1993/94 to follow changes in size and age structure of the clam populations in different parts of the study area. Large spatial differences in population structure were found between the sheltered southwest of the Bay and the shallow and exposed Oder Bank in the centre. The stock of the Oder Bank consisted of two different clam types. A slow-growing cohort was assumed to be autochtonous on the Oder Bank, whereas a fast-growing one was assumed to have immigrated from the surrounding area. The contribution of the two cohorts to the total density varied seasonally. Because of bedload transport of clams, the contribution to the reduction of the clam stock by wintering sea ducks could not be quantified. In the southwest of the Pomeranian Bay erosion was of minor importance. High mortality rates during the first two years of life were assumed to be caused by predation. Mortality rates of older cohorts remained stable until old age. Variations in cohort strength were related to interannual differences in the reproductive success. A mild winter presumably lowers the reproductive success in the subsequent summer. Severe oxygen depletion in summer 1994 caused a strong reduction in the clam stock at stations deeper than 10 m.     

菲律宾蛤仔(Ruditapes philippinarum)金属硫蛋白基因克隆及Cd~(2+)胁迫下的表达分析

金属硫蛋白(Metallothionein,MT)是一类富含半胱氨酸的小分子蛋白质,参与机体重金属解毒和金属元素代谢等生理过程。本研究采用RACE技术,克隆获得了菲律宾蛤仔(Ruditapes philippinarum)金属硫蛋白(RpMT)的全长cDNA序列。RpMT的cDNA全长为570bp,编码75个氨基酸,包含15个MT所特有的Cys-Xn-Cys结构。采用实时荧光定量PCR技术,分析了两种壳色菲律宾蛤仔(白蛤和斑马蛤)RpMT基因在Cd2+暴露后的表达变化。结果发现:Cd2+急性和亚慢性暴露均可导致两种壳色蛤仔消化腺和鳃组织RpMT基因表达量的显著上调;暴露后两种壳色蛤仔鳃组织RpMT基因表达量的增加幅度均高于消化腺组织,且以白蛤鳃组织基因表达水平的上调幅度较高。上述结果表明,RpMT可能在菲律宾蛤仔抵御Cd2+胁迫过程中发挥了重要作用。     

Biochemical and histochemical responses to environmental contaminants in clam, Tapes philippinarum, transplanted to different polluted areas of Venice Lagoon, Italy.

In this study the clam, Tapes philippinarum, a filter-feeding bivalve living in soft bottoms, was used to investigate the water/sediment pollution in Venice Lagoon (Venice, Italy), a heavily urbanised and industrialised area. To this end, clams collected from indigenous populations in a clean farming area (Val Dogà) were transplanted into a relatively low contaminated site (Palude della Rosa) and in a heavily polluted area (Porto Marghera), for 5 weeks. A range of possible induced stress indices were measured, on the basis of potential toxicity mechanisms, including specific enzymatic activities [Bap hydroxylase and NADPH cytochrome c reductase, superoxide dismutase, catalase, glutathione peroxidase and propionaldehyde dehydrogenase (PDH)] and general cellular responses (lysosomal latency). Moreover, chemical analyses (polychlorinated biphenyls, polycyclic aromatic hydrocarbon, DDTs) were carried out on sediments and animal tissues at the beginning and end of the transplantation experiments. The chemical results show an active bio-accumulation activity from sediment in clams translocated to the most polluted area. Correspondingly, biological data indicate marked effects on PDH activity and latency in T. philippinarum from the same site. In contrast, no similar pattern has been observed in the other considered indices, possibly due to antagonistic effects of the complex contaminant mixture present in the environment.     

哈维氏弧菌对泥蚶的毒性及半致死浓度研究

为构建泥蚶感染弧菌标准技术体系,本研究采用哈维氏弧菌浸泡感染和注射胁迫两种攻毒方式对泥蚶进行了急性毒性实验,观察泥蚶在菌液胁迫下的存活情况,分析哈维氏弧菌对泥蚶的半致死浓度。浸泡感染实验结果表明,随着感染水体中弧菌升高,泥蚶死亡起始时间和最大日死亡率时间均有所提前,泥蚶死亡情况呈现上升趋势,当浸泡浓度超过2×10⁷CFU/mL,泥蚶15 d累计死亡率接近100%。注射胁迫实验结果表明,泥蚶死亡率与弧菌注射浓度呈显著正相关(r=0.964,P<0.01)。Probit回归分析结果显示,哈维氏弧菌浸泡感染泥蚶7 d后停止实验,15 d的半致死浓度(LC₅₀)为1.35×10⁷CFU/mL;注射胁迫下的96 h半致死剂量(96 h LD₅₀)为2.12×10⁷CFU/g,说明哈维氏弧菌对泥蚶有明显的毒害作用。研究结果为泥蚶对哈维氏弧菌耐受性提供数据参考,并且为泥蚶弧菌病防治提供了理论基础。     

Infection by gymnophallid metacercariae enhances predation mortality of SW Atlantic stout razor clam Tagelus plebeius

Parasite life cycles are frequently completed in different hosts, thus the parasites have its life cycle overlapped to natural trophic webs. The family Gymnophallidae (Class: Trematoda; Subclass: Digenea) includes digenetic parasites whose larval stages occur on bivalves and may affect bivalve predation by the final host of these parasites. In this work we evaluated: (a) if individuals of the razor clam Tagelus plebeius with higher parasite intensity suffer higher predation by the oystercatcher Haematopus palliatus and, (b) if there is any effect of parasite intensity on burrowing and escape behaviours of these razor clams which may enhance exposure to predators. Field experiments (oystercatcher exclusion vs. open access) showed that clams with higher parasite intensity support higher predation by oystercatchers, which suggests a higher consumption of more parasitized clams and thus, a more successful reproduction of parasites linked to the intensity of infection. However, clam burrowing and escape behaviours did not show differences related to different parasite intensity, suggesting that the commonly believed mechanisms are not responsible in this case.     

养殖迁移对菲律宾蛤仔挥发性有机物质组成变化的影响

环境条件变化是造成贝类挥发性有机物质(VOCs)组成变化的主要原因。菲律宾蛤仔养殖生产中,常进行从开放式自然滩涂迁移至人工围塘养殖的活动。本文研究了这一过程前后菲律宾蛤仔(Ruditapes philippinarum)肌肉组织的VOCs组成特征、变化及其与环境因子相关性。结果显示:基于固相微萃取和气质联用技术,共鉴定出52种VOCs,分别属于醛、醇、酯、烃、呋喃、烷烃、烯、芳香烃、含硫和含氮化合物等。迁移前样本中,检出41种化合物,迁移至人工围塘养殖20 d后,检出32种。迁移造成了化合物种类数量降低、种类发生变化。醛、醇类的相对含量最高、迁移前后的变化尤为显著,呈现由醛高醇低的组成逆转为醇高醛低的特点。溶氧是影响VOCs组成变化及其醇、醛类差别的重要因子。饵料生物量、相对丰度和多样性与VOCs的组成具有显著相关性:自然滩涂微型真核生物多样性高但生物量和丰度相对低,人工围塘多样性低但生物量和丰度高,是VOCs组成差异的关键影响因子。本研究建立了贝类VOCs与关键环境因子的直接联系,为进一步解析贝类品质的环境关系、优化和调控养殖环境等研究提供了参考。     

Field assessment of biochemical stress indices for the sandworm Neanthes virens (Sars)

Certain biochemical stress responses have been observed in the laboratory for Neathhes virens subjected to various environmental and pollutant stressors. The purpose of the present study was to determine whether similar alterations in these biochemical parameters could be detected in a natural population of N. virens from a contaminated site as compared with animals from a nearby reference site. A site in Portland Harbor, Maine, adjacent to an oil storage facility with oil-contaminated sediments was selected as the experimental study area and the reference site was at Pine Point, approximately 15 miles south of Portland.Animals were collected from the two sites on consecutive days on a bimonthly basis at low tide within a day of the full moon from September 1981 through March 1982. Coelomic fluid samples were taken within 2 h after collection and the animals were then frozen in liquid nitrogen for transport back to the laboratory. Coelomic fluid samples were analyzed for glucose and the frozen tissues were analyzed for glycogen, lipid and free amino acid concentrations.Significant differences in these parameters were often observed between the two populations. The glycogen content of animals from the contaminated site was always less than that of animals from the reference site, whereas the lipid levels were always higher. Significant differences between the free amino acid composition of the animals from the two sites were also observed. Although distinct biochemical differences can be detected between these two populations, the considerable biological variation in these parameters may limit their usefulness for field monitoring investigations for this species.     

Response of antioxidant systems to copper in the gills of the clam Ruditapes decussatus

Copper (Cu) is an essential element for biological systems, however, when present in excess, is toxic. Metallothioneins can play an important role in Cu homeostasis and detoxification. Moreover, Cu can catalyse the production of toxic hydroxyl radicals that cause lipid peroxidation but defence systems in the cells can limit the oxidative damage. The present study was performed to investigate the effect of three Cu concentrations (0.5, 2.5 and 25 μg l⁻¹) on the response of antioxidant enzyme activities (superoxide dismutase (SOD), catalase (CAT), selenium-dependent glutathion peroxidase and total glutathion peroxidase), total proteins, metallothioneins (MT), malondialdehyde (MDA) concentrations in the gills of the clam, Ruditapes decussatus. The activity of antioxidant enzymes and total proteins, MT and MDA concentrations were measured in the gills of the clams after 1, 3, 7, 14, 21 and 28 days of Cu exposure. Results indicate that Cu only induces an imbalance in the oxygen metabolism during the first week of Cu exposure due to a decrease in mitochondrial SOD and CAT, selenium-dependent and total glutathion peroxidase activities. Cu also causes lipid peroxidation, measured by the MDA formation, that was Cu dependent. In the gills of clams exposed to 25 μg Cu/l, the excess of Cu triggers the induction of MT synthesis after 3 days of exposure.     

Bioavailability of heavy metals bound to sediments affected by a mining spill using Solea senegalensis and Scrobicularia plana

After the Aznalcóllar mining spill (April, 1998) different ecosystems along the Guadiamar river and the Guadalquivir estuary were impacted by high concentrations of metals. The concentration of metals Zn, Cd, Pb, Cu and As has been detected as a good tracer of the pathway of the toxic spill. Different individuals of the fish Solea senegalensis and the clam Scrobicularia plana were exposed to contaminated sediments after the accidental spill in the south-west Spain. The induction of metallothioneins in fish and the bioaccumulation of metals in clams were determined in different tissues to determine the biological adverse effect associated with the heavy metals from the spill. A relationship between the enrichment of metals in the tissues and the induction of metallothioneins was determined for those assays performed using contaminated sediments. The bioaccumulation of metals indicates that the risk associated with the spill should be continuously evaluated to establish the sublethal effects related to the accident.     

Vertical zonation and aggregated distribution of the Manila clam on subtidal sand flats in a coastal brackish lagoon along the Sea of Japan

The Manila clam, Ruditapes philippinarum, has maintained small‐sized populations in a semi‐enclosed brackish lake along the Sea of Japan, the Honjo area of Lake Nakaumi, although the environment and biota of this area have changed dramatically due to a large‐scale reclamation project. There should be underlying processes that enable the restoration of this species from small‐sized populations, such as the existence of source (i.e. reproductive) populations in other areas and depth zones of the lake. However, there has been no robust, properly designed evaluation of the distribution of the Manila clam in the subtidal sand flats. In order to elucidate the possible mechanisms that allow for the persistence of populations of the Manila clam, we examined the spatiotemporal and vertical variation in distributions of 0‐age clams in the subtidal zone of sand flats. Seasonal effects on population variations showed erratic changes among depth zones without a decreasing trend along the depth gradient. Further, many local populations became extinct even in the shallower zones due to seasonal (summer) hypoxia at deeper zones and hypoxia by the accumulation of key benthic species (Asian mussel and decaying macroalgae) in mats at shallower zones. A few surviving local populations were stable with a spatial‐fragmental (patchy) distribution, associated with fragmented accumulations of Asian mussels and macroalgae. Efforts to maintain stable populations and to restore this species in the subtidal area may depend on these small, restricted, patchy local populations. These findings suggest that high fertility and productivity of the Manila clam as well as patchy distribution of small populations may contribute to the maintenance of the population and the avoidance of extinction (by spatially diffusing the risk of extinction) in harsh environments resulting from the reclamation project.