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(P0.05),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(P0.05);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(P0.05);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(P0.05),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(P0.05),推测TYR基因在五个菲律宾蛤仔群体的鳃和肝胰腺中参与了免疫应答。通过对TYR基因的氨基酸序列进行二级结构分析和系统发育树分析,找到两个铜离子结合位点和6个组氨酸残基,并发现TYR6基因和长牡蛎同源性最高,为39.43%, TYR10基因和大珠母贝同源性最高,为51.04%。本文首次探讨在LPS胁迫下菲律宾蛤仔TYR基因的表达特性,为进一步探究TYR基因与菲律宾蛤仔免疫应答机制提供参考。     

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。     

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...     

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.     

Clam community composition and prey shell size impacts moon snail (Gastropod: Naticidae) drilling frequencies in South Carolina,USA

Moon snail predation on clams is a common model system of predator–prey interactions. In this system, the predator bores through the shell of its prey, leaving a distinct and identifiable hole. Some paleoecological and behavioral research on moon snails suggests a trend in predation preference directed toward clams with small shells. Rarely, however, have studies tested relative drilling frequencies across species and size ranges in natural assemblages of clam communities. We examined the clam community composition at two beaches in South Carolina, USA, and we then tested moon snail predator preferences for (a) clam prey species and (b) whether their selection is related to prey shell size. We collected a total of 1,879 clam shells, identified each shell to species and recorded their anteroposterior length. The species composition of clams differed significantly between the two beaches; Anadara ovalis was dominant at both sites, but three of ten total species were only collected at one beach. Folly Beach had nearly a 60% higher the overall drilling frequency (34.6%) versus Edisto Beach (21.8%), and this may be linked to the differences in clam community compositions at the sites. For A. ovalis and Mulinia lateralis, shells with larger lengths have lower probabilities of being bored by a moon snail. Anadara brasiliana, which generally is a thinner‐shelled clam species, had the highest total drilling frequency (77.2%), and Noetia ponderosa, a thicker‐shelled clam, had a considerably lower drilling frequency (12.0%). We conclude that both community level factors (species composition) and population characteristics (shell size distributions) may influence the local drilling frequency by moon snails.     

砗蚝(Hippopus hippopus)的人工繁育

于2016年4—9月开展了砗蚝(Hippopus hippopus)人工繁育技术研究。采用五羟色胺进行催产、促使配子排放;精卵分别收集,进行异体间受精以避免自交;受精卵经过30h孵化,选出600万D形幼虫进行培育。砗蚝的早期生活史与砗磲(Tridacna spp.)相似,经历前期面盘幼虫、中期面盘幼虫、后期面盘幼虫、足面盘幼虫、单水管稚贝、双水管稚贝、外套膜触手稚贝、幼贝等阶段。与砗磲不同的是,砗蚝怀卵量较少,但卵径较大,D型幼虫也较大,幼虫趋光性更强,壳长2.0mm以后外套膜不伸出壳缘外,幼贝贝壳形态也不同于砗磲。同砗磲幼虫一样,砗蚝幼虫需要构建虫黄藻系统之后,才能出现鳃、次生壳等,从而完成变态成为稚贝。砗蚝幼虫变态率较低,仅为1.4%。中间育成期间,丝状藻和锥形螺是稚贝培育的主要敌害,需及时清理才能确保稚幼贝正常生长发育。经过120d的精心饲育,培养出平均壳长6.3mm幼贝500余个。本研究为进一步开展砗蚝人工繁育、中间育成、增殖放流、资源修复及移植保育提供了参考。     

Feeding ecology of whelks on an intertidal sand flat in north‐eastern New Zealand

The dispersion patterns and feeding behaviour of intertidal whelks were investigated in north‐eastern New Zealand. Aggregations of whelks feeding on clams occurred regularly, with an average of 15–20 individuals per aggregation. Death of clams was attributable to predation in up to 50% of the aggregations, although scavenging of dead and moribund animals was the main activity of whelks. Laboratory and field studies showed that Lepsiella scobina (generally not considered a soft shore species) was the main whelk predator at Lews Bay, Whangateau Harbour, drilling a small neat hole through the clam shell. The distinction between Cominella spp. as predators or scavengers is not so clear. Reseeding of intertidal clams, Austrovenus stutchburyi, has been proposed as a technique for reestablishing populations in some degraded New Zealand estuaries. The impact of whelk predation has many implications for A. stutchburyi reseeding. Both L. scobina and Cominella adspersa may potentially prey on newly reseeded clams as they preferentially attacked small clams in the laboratory. On‐growing to a larger size before reseeding may be advantageous although large size was not found to protect prey from predation. Additionally, no seasonal trend in whelk activity was found.     

基于GLM模型Logistic回归的菲律宾蛤仔地理群体判别

菲律宾蛤仔(Ruditapesphilippinarum)是重要的海洋经济贝类和海水养殖品种,具有较强的表型可塑性特点,不同地理群体之间存在表型差异。本研究根据实测数据建立了胶州湾与天鹅湖菲律宾蛤仔壳长(SL)与壳高(SH)、壳宽(SW)和活体重(W)的回归方程,定量分析了两个地理群体的形态学差异。由于其近似圆形的形态使得所能测量的外部指标较少,限制了传统形态测量学在其地理群体判别中的应用。本研究提出了将传统形态测量学和广义线性模型(GLM)相结合的方法,建立了判别菲律宾蛤仔地理群体的逻辑斯蒂(Logistic)回归方程。结果表明:菲律宾蛤仔的天鹅湖群体比胶州湾群体具有更高的SH/SL和SW/SL比值,壳形椭圆较圆且隆起程度高,"凸"形明显;基于GLM模型的Logistic回归能够快速准确地判别菲律宾蛤仔的地理群体归属,判别正确率高达94.23%。本研究为双壳类物种地理群体判别提供了新的分析方法和技术手段。     

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。     

Preliminary results of a field study on some stress-related parameters in Tapes philippinarum naturally infected by the protozoan Perkinsus sp.

The aim of the present work was to investigate the possible use of the clam Tapes philippinarum—a filter-feeding bivalve living on soft bottoms—in the monitoring of sediment pollution in the Lagoon of Venice (North-East Italy). To this end, two clam populations, exposed to different environmental conditions (a relative clean site and an industrial polluted area), have been monitored for the prevalence of the protozoan parasite Perkinsus sp. as well as for several biological parameters commonly used as determinants of contaminants exposure. First results showed a low prevalence, higher in the clean site, but a very diffuse distribution of the parasite throughout the two populations. Nevertheless, greater digestive cells height and longer lysosomal latency time were observed in clams from the clean area. As for the physiological stress-related parameters, even if clearance rates were always higher in these clams, scope for growth values indicated a more complicated biological response.     

<Emphasis Type="Italic">Zostera marina</Emphasis> seed burial can be enhanced by Manila clam <Emphasis Type="Italic">Ruditapes philippinarum</Emphasis>: A microcosm study

Seagrass seed bank plays a key role in the regeneration of new vegetation when seagrasses are removed by the natural or man-made disaster. Various factors may affect the development of sediment seed bank. We conducted a microcosm experiment to test the effects of burrowing and feeding activities of Manila clam, Ruditapes philippinarum on the burial of Zostera marina seeds in sediments. The effects of lasting time (3-hour, 1-day, 3-day, 7-day, 14-day and 28-day), clam density (0, 2, 4 and 8 clams with shell length of 3 cm in each microcosm) and clam size (shell length of 2, 3 and 4 cm at 4-clam density) on seed burial were examined in plastic microcosm cores (30 cm high × 10 in inner diameter) in a 28-day period. Results showed that the seed burial depth significantly increased with time, the density and the size of clams. No seeds were buried in the sediment in the cores without clams during the whole experiment period. For the 3-cm clams, about 91.61% of the seeds were buried in the sediment at the end of the experiment in the high-density treatment (8 clams at each core); while in the medium and low-density treatments (4 and 2 clams in each core, respectively), about 76.93% and 60.61% of the seeds were buried in the sediment, respectively. For the size treatments, large (4 cm) clams buried 89.56% of the seeds at the end of the experiment, much more than those of medium (3 cm, 76.93%) and small (2 cm, 61.50%) size clams. During the whole experiment period, nearly all of the buried seeds were at a depth of from 0 cm to 5 cm. These results suggested that Manila clam Ruditapes philippinarum may play an important positive role in seagrass seed bank dynamics in the field.     

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。     

辽宁沿海菲律宾蛤仔不同地理群体形态差异研究

利用多变量形态度量学方法,对辽宁沿海菲律宾蛤仔(Ruditapes philippinarum)长兴岛(CXD)、旅顺(LS)、营口(YK)、盘锦(PJ)、锦州(JZH)、海洋岛(HYD)、黑石礁(HSJ)以及金州(JZ)8个野生群体进行形态学比较分析。结果表明:主成分分析得到了3个主成分,贡献率分别为40.21%,19.98%和11.57%,累积贡献率为71.75%。聚类分析结果显示,旅顺群体和锦州群体形态最为接近,而与黑石礁群体差异较大。营口群体的趋异程度最大,长兴岛群体、金州群体和海洋岛群体的趋异程度居中。长兴岛群体壳型最扁,贝壳较厚且重;营口群体软体部最肥;金州群体壳型"凸"型最明显,且质量最大,而黑石礁群体贝壳最薄。这些明显特征揭示了不同群体的形态差异。研究结果为菲律宾蛤仔的地理群体识别、种质资源保护与恢复提供了重要依据。     

Changes in survivorship,behavior, and morphology in native soft‐shell clams induced by invasive green crab predators

Many studies on invasive species show reduced native densities, but few studies measure trait‐mediated effects as mechanisms for changes in native growth rates and population dynamics. Where native prey face invasive predators, mechanisms for phenotypic change include selective predation, or induced behavioral or morphological plasticity. Invasive green crabs, Carcinus maenas, have contributed to declines in native soft‐shell clams, Mya arenaria, in coastal New England, USA. We tested the hypothesis that clam ability to detect chemical cues from predators or damaged conspecifics would induce greater burrowing depth as a refuge from invasive crabs, and greater burrowing would require increased siphon growth. To determine how crab predation affected clam survivorship and phenotypic traits in the field, clams in exclosure, open, and crab enclosure plots were compared. Crab predation reduced clam density, and surviving clams were deeper and larger, with longer siphons. To determine whether the mechanism for these results was selective predation or induced plasticity, phenotypes were compared between clams exposed to chemical cues from crab predation and clams exposed to seawater in laboratory and field experiments. In response to crab predation cues, clams burrowed deeper, with longer siphons and greater siphon mass. Overall, crab predation removed clams with shorter siphons at shallow depths, and crab predation cues induced greater burrowing depths and longer siphons. Longer siphons and greater siphon mass of deeper clams suggests clams may allocate energy to siphon growth in response to crabs. By determining native behavior and morphological changes in response to an invasive predator, this study adds to our understanding of mechanisms for invasive impacts and illustrates the utility of measuring trait‐mediated effects to investigate predator–prey dynamics.     

Influence of nutritional stress on digestive enzyme activities in juveniles of two marine clam species, Ruditapes decussatus and Venerupis pullastra

The potential use of digestive activities as indicators of the nutritional status in bivalves is discussed in relation to the results obtained in two clam species exposed to starvation and refeeding. Activities of some digestive enzymes (amylase, laminarinase, cellulase, and protease) were measured in juveniles of two commercially interesting species of clams, Ruditapes decussatus and Venerupis pullastra. The specimens were fed normally, being after subjected to a 15-days starvation and a further refeeding period. Samples were obtained at different moments of such feeding schedule to evaluate enzymes as well as weight (live, dry and organic) and length, in order to calculate growth rates and feeding efficiencies. Starvation led to a major decrease in clam growth as measured by dry weight and a negative growth as measured by organic weight, this coinciding with a certain degree of growth of the shell and a consumption of soft tissue. This response occurred more rapidly in R. decussatus but was of a lower magnitude than in V. pullastra. Activity of carbohydrases decreased rapidly in both species with starvation, although protease activity was maintained in R. decussatus. Recovery after the end of starvation was not similar in both species; while R. decussatus attained similar growth rates and enzyme activities to those measured prior to nutritional stress, V. pullastra only recovered 50% of its initial values. For both species of bivalves it can be concluded that digestive enzymes, and more specifically amylase, could be used as indicative of their nutritional condition.     

中国蛤蜊Mactra chinensis Philippi稚贝运动行为学初步研究

为从行为学角度解释中国蛤蜊(Mactra chinensis Philippi)的跑滩现象, 研究了规格、温度对其跳跃行为以及底质、流速和周期性干露对其潜沙行为的影响。结果表明, 中国蛤蜊运动能力与其规格和水温有关, 3 mm稚贝不具备跳跃能力, 5 mm以上具备跳跃能力; 跳跃频率和高度随水温的升高呈上升趋势。中国蛤蜊稚贝潜沙比例均随时间推移逐渐上升; 各时间节点中国蛤蜊稚贝在细沙底质的潜沙比例最高, 中沙底质次之, 粗沙底质最低。施加微弱水流后, 中国蛤蜊稚贝潜沙速度和比例显著增加; 流速增大到一定程度后部分稚贝被水流冲走, 潜沙比例随之下降。周期性干露会导致中国蛤蜊稚贝从底质中爬出, 干露时间越长, 爬出比例越高。干露后的稚贝重新放置于海水中, 部分稚贝会在水面漂浮一段时间后再下落至水底, 漂浮比例随干露时间的增加而增加。本研究从行为学角度初步查明了中国蛤蜊跑滩现象, 为解决中国蛤蜊防跑滩问题提供了思路。     

人工养殖曼氏无针乌贼(Sepiella maindroni)繁殖生物学特性研究

采用常规生物学方法和统计回归分析,研究了养殖曼氏无针乌贼卵巢成熟系数(GSI)的变化规律和个体生殖力与其主要形态指标之间的相互关系.结果表明,曼氏无针乌贼春苗100日龄时GS/达到最高(12.2%±3.25%),秋苗140日龄时GSI达到最高(13.7%±3.69%).曼氏无针乌贼秋苗的体质量相对生殖力(F<,W>)极...     

浙江近海曼氏无针乌贼(Sepiella maindroni) 繁殖生物学特性变化研究

采用常规生物学方法和统计回归分析方法,对浙江近海曼氏无针乌贼的繁殖生物学特性进行了研究,比较了不同时期浙江近海曼氏无针乌贼卵巢成熟系数(GSI)、生物学体型及个体生殖力与生物学指标关系。结果表明,浙江近海曼氏无针乌贼GSI在4月份达到最高(9.51%±7.23%),相比20世纪60年代初、80年代初,自然海域曼氏无针乌贼的性腺发育提前,且生殖期延长。20世纪80年代初曼氏无针乌贼的生物学体型、怀卵量及个体生殖力均大于60年代初和自然海域曼氏无针乌贼,同时曼氏无针乌贼在人工养殖条件下生物学体型、怀卵量及个体生殖力增大。浙江近海曼氏无针乌贼的个体生殖力均随着胴长(ML)、体质量(W)、卵巢质量(WO)、GSI这些指标的增大而增大,除体质量相对生殖力(FW)与ML为密切正相关外,个体生殖力与这些指标的关系均达到了极显著水平(P<0.01)。多元线性逐步回归分析表明,WO是影响浙江近海曼氏无针乌贼个体生殖力的最重要指标。     

A comparison of condition factor and gonadosomatic index of sardine Sardinops sagax stocks in the northern and southern Benguela upwelling ecosystems, 1984–1999

Time-series of condition factor (CF) and gonadosomatic index (GSI) were generated using general linear models (GLM) for sardine Sardinops sagax stocks in the northern and southern Benguela ecosystems over the period 1984–1999. During this period the biomass of sardine in the northern Benguela remained at relatively low levels of <500 000 tons, whereas that of southern Benguela sardine increased 40-fold to 1.3 million tons. The GLMs explained 27 and 45% of the observed variation in CF, and 32 and 28% of the observed variation in GSI, for sardine in the northern and southern Benguela subsystems respectively. Whereas the sardine CF in the northern Benguela remained stable over time, that for the southern Benguela stock declined steadily during the study period. Sardine CF showed a seasonal cycle in the southern but not in the northern Benguela. Time-series of GSI showed high interannual variability but no trends in either subsystem, and the seasonal pattern was similar for both stocks. The lack of coherence between the CF time-series for sardine in the two subsystems further suggests that sardine stocks in the northern and southern Benguela subsystems are independent.     

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.