Effect of reproductive toxicants on lipovitellin in female blue crabs, Callinectes sapidus

Changes in the concentration of lipovitellin, a lipoprotein that is the major protein in mature oocytes of crustaceans, were determined during ovarian development in female blue crabs, Callinectes sapidus. To test the effects of reproductive toxins on vitellogenesis, crabs were fed food containing cadmium or injected with the juvenile hormone antagonist, precocene II. Monoclonal antibodies prepared against one of the lipovitellin peptides (peptide A—107 kDa) were used in an enzyme linked immunosorbent assay to determine lipovitellin concentrations. Recently molted female crabs fed food containing cadmium (0.02 mg/g) for 18 days had oocytes with the same lipovitellin concentration as controls, but after 24 days the oocytes in cadmium treated crabs had significantly lower lipoprotein contents than controls (30 ± 4 ng/oocyte—cadmium treated; 45 ± 4 ng/oocyte—control). Precocene II-treated crabs had significantiy lower lipovitellin concentrations after 18 and 24 days. Diameters of oocytes were the same in both treated and control crabs. The results suggest that some pollutants may affect reproduction by acting during secondary vitellogenesis i.e., period when there is a large increase in lipovitellin synthesis. Since lipovitellins serve many of the nutritional needs of recently emerged crab embryos, poor larval survival may occur as a result of lower lipovitellin content.     

Movement patterns and trajectories of ovigerous blue crabs Callinectes sapidus during the spawning migration

Female blue crabs (Callinectes sapidus Rathbun) migrate from low salinity estuarine regions to high salinity regions near the ocean to release larvae. During this migration, ovigerous females use ebb-tide transport, a vertical migratory behavior in which they ascend into the water column during ebb tides, to move seaward to larval release areas. In order to determine the relationship of ebb-tide vertical migrations to local currents and the influence of these vertical migrations on the horizontal transport of blue crabs in the estuary, ovigerous females with mature embryos (1–3 days from hatching) were tracked near Beaufort Inlet, North Carolina (USA), in July and August 2001 and 2002. Crabs were tagged and tracked using ultrasonic telemetry, and currents near the crabs were measured simultaneously with a shipboard acoustic Doppler current profiler.During the two seasons, eight crabs were successfully tracked for periods ranging from 3.9–37.0 h and for distances ranging from 1.9–10.6 km. All crabs migrated seaward during the tracking periods. Crabs moved episodically during all tidal phases with periods of movement on the order of minutes to an hour. They moved with local currents in terms of both speed and direction during ebb tides, consistent with ebb-tide transport, and moved down-estuary (seaward) in opposition to local currents during flood tides. The percentage of time that crabs were active was higher during night ebb tides than during day ebb tides or flood tides and increased with increasing ebb-tide current speed. Mean migratory speeds were 0.11, 0.04, 0.08 and 0.02 m s⁻¹ during night ebb, night flood, day ebb and day flood tides, respectively, and net migratory speeds were on the order of 5 km day⁻¹. Due to the episodic nature of the crabs' movements, the total distances that crabs traveled during ebb tides ranged from 10–40% of the distances that passive particles could have traveled under the same conditions.     

墨西哥湾海洋环境对蓝蟹(Callinectes sapidus)幼体分布和扩散的影响

本文以蓝蟹为例,研究海洋环境对甲壳动物幼体迁移规律和机制的影响。利用不规则三角形网格和有限体积模型(finite-volume coastal ocean model, FVCOM)耦合kinesis模型的方法,分析研究了墨西哥湾物理环境对蓝蟹(Callinectessapidus)幼体的分布和扩散途径的影响。蓝蟹在每年的四、五月份海水落潮期间产卵,通过跟踪算法从产卵区域沿着墨西哥湾海域进行模拟,获得了80天内蓝蟹幼体的粒子移动轨迹,记录并分析了幼体经过海域的盐度值。研究结果证明了该方法可有效模拟蓝蟹幼体在特定海域的迁移规律和扩散机制,进一步研究可为了解海洋物理环境对蓝蟹和其他渔业资源的影响提供借鉴。     

Cadmium-binding proteins in the blue crab, callinectes sapidus: laboratory-field comparison

The blue crab, Callinectes sapidus, is distributed along the east coast of the United States from Cape Cod, Mass., through the Gulf of Mexico, including both relatively unpolluted coastal areas and estuaries contaminated with trace metals. Cadmium is of particular concern because it is concentrated in the digestive glands of blue crabs and can be passed on to consumer organisms. Tissue concentrations and partitioning of trace metals from crabs exposed in the laboratory to 10 ppb dissolved cadmium for 40 days were compared with blue crabs collected from two locations on the Hudson River, NY, Foundry Cove and Haverstraw Bay, Foundry Cove and Haverstraw Bay, both of which have elevated trace metal levels relative to estuarine areas near Beaufort, NC. Crab digestive glands, gills and muscle were removed and analyzed for total cadmium, copper, zinc and nickel concentrations using acid digestion and atomic absorption spectrophotometry, and metal-binding (metallothionein-like) proteins were determined by gel-filtration chromatography. In crabs exposed to cadmium in the laboratory, the cytosolic partitioning was similar to previous investigations at our laboratory where higher levels of cadmium (100 ppb) and shorter exposure times (14 days) were used. The similarity in cadmium partitioning from these two separate experiments indicates dose independence. In crabs from polluted environments the digestive glands contained the highest concentrations of trace metals. Chromatograms of the cytosol from the digestive glands and gills from both field and laboratory exposed crabs showed similar distributions of cadmium, copper and zinc. The gills of both field and laboratory-exposed crabs had metal-binding proteins that contained mostly cadmium, and the digestive glands had metallothionein-like proteins that contained cadmium, copper and zinc. Estimated molecular weights for these proteins were similar to the metallothioneins found in other crustaceans and mammals.     

Cadmium accumulation by the blue crab, callinectes sapidus: Involvement of hemocyanin and characterization of cadmium-binding proteins

The accumulation of cadmium from seawater by the blue crab, Callinectes sapidus, was studied as a function of metal concentration and exposure time, with special emphasis on cadmium-binding proteins. Cadmium was found, in decreasing order of magnitude, in gills, digestive gland and hemolymph. When exposed to 0·5 ppm cadmium for 2–24h, virtually all of the cadmium in the cytosolic fraction of the gill was associated with a low molecular weight (LMW) cadmium-binding protein (MW 8000). However, after 48h of exposure only 50% of the cadmium in the cytosol was bound to this protein. The rest wasfound to be associated with proteins of a molecular weight of 300 000 and 60 000. This pattern of cadmium distribution did not change over a 12-day depuration period. Similar results were obtained upon exposure to 0·1 ppm cadmium. The pattern of cadmium accumulation in the cytosolic fraction of the digestive gland was in marked contrast to that observed for the gill. Initially, the cadmium was distributed over three low molecular weight fractions. During depuration the distribution of cadmium changed and all of the metal became bound to a low molecular weight protein (MW 9000). The cadmium concentrations in the gill and digestive gland remained essentially constant during depuration (12 days). The LMW cadmium-binding proteins were purified by a combination of gelpermeation and ion-exchange chromatography. Their molecular weight, spectral properties and amino acid composition are characteristic of the vertebrate metallothioneins. During exposure to cadmium the metal rapidly appeared in the hemolymph, mainly associated with hemocyanin. During depuration cadmium was transferred from the hemolymph to the digestive gland, demonstrating that hemocyanin acts as a carrier in trace metal transport.     

Detection of the water-soluble fraction of crude oil by the blue crab, Callinectes sapidus

The ability of the blue crab, Callinectes sapidus, to detect petroleum hydrocarbons was measured with behavioural techniques. When presented with a water-soluble fraction of Prudhoe Bay crude oil, blue crabs abruptly changed antennular orientation, began rhythmic beating of the maxillipedal flagellae, and increased antennular flicking rate. The threshold concentration at which 50% of the crabs detected the water-soluble fraction was 2 × 10^?6 mg/litre. The blue crab apparently can readily detect petroleum hydrocarbons at concentrations found in chronically polluted areas as well as oil spill situations.     

三疣梭子蟹精子保存研究

于2004年5月到2004年7月对三疣梭子蟹精子进行了常温、低温和超低温保存研究。利用离子载体A23187人工诱导顶体反应以判断精子活力。常温和低温保存实验的结果显示,4℃下无钙人工海水(calcium-free artificial sea water,Ca2 FASW)保存236h的精子有25%以上的成活率。在4℃,25℃两种温度与Ca2 FASW、天然海水(natural sea water,NSW)两种基础液构成的4种组合中,最优组为4℃、Ca2 FASW,最差组为25℃、NSW。将精子浓度与Ca2 FASW对照组及诱导组比较,结果显示,25℃和4℃下,精子浓度与对照组顶体反应率相关性不显著(P=0.068>0.05,P=0.265>0.05),与诱导组顶体反应率极显著相关(P<0.01)。以二甲亚砜(Dimethyl sulfoxide,DMSO)和甘油(Glvcerol,G)作为抗冻剂进行超低温冷冻实验,72h后解冻,包括对照组在内的所有组均有98%以上的精子不经诱导就发生顶体反应。     

Heavy metal stress induced hyperglycemia in blue swimmer crab, Portunus pelagicus

The study was to find out the effect of cadmium and mercury on levels of hemolymph glucose, crustacean hyperglycemic hormone (CHH) and hepatopancreas glycogen in the blue swimmer crab Portunus pelagicus. The experiments were performed in both intact and eyestalk ablated crabs. Quantification of CHH was done by the indirect ELISA with the aid of primary anti-Carcinus maenas-CHH antibody. Higher glucose concentration was observed on exposure to 8×10^-6 of cadmium ((825.6±5.42) μg/mL) and 6×10^-6 of mercury ((90.5±6.25) μg/mL) after 48 h and 24 h respectively. Higher level of hemolymph glucose was observed in eyestalk intact crabs on exposure to cadmium and mercury than eyestalk ablated crabs. Decrease in the levels of CHH was observed in both eyestalk intact and ablated crabs on heavy metal exposure. Decline of the hepatopancreas glycogen level was also witnessed with the exposure to heavy metal, which validated its utilization in the production of glucose. Thus this study brings to light, the variations in hemolymph glucose, CHH and hepatopancreas glycogen on heavy metal stress. These carbohydrate metabolites can be used as biomarkers in assessing heavy metal contamination in water bodies.     

Effects of organic toxicants on the anoxic energy metabolism of the mussel Mytilus edulis

The anoxic rates of heat dissipation by mussels (Mytilus edulis) were enhanced after exposure to pentachlorophenol (PCP) and tributyltin (TBT), both known uncouplers of oxidative phosphorylation. The degree of metabolic activation under anoxia was dependent upon the amount of toxicant accumulated in the tissues, but the concentration-response relationship was different to that under aerobic conditions. Biochemical measurements indicated that the anaerobic metabolic pathways were significantly disturbed by PCP and TBT. There was a decline in succinate, an increase in the fumarate: succinate ratio, and an increase in the accumulation of lactate, indicating a shift from succinate to lactate anaerobic pathways. A consequence of organic toxicant exposure was a reduction in anoxic survival time of mussels.     

维生素C、E对镉致毒后中华绒螯蟹抗氧化系统酶活性的影响

通过在饲料中添加不同浓度的维生素C(VC),维生素E(VE)探讨其对镉致毒后中华绒螯蟹(Eriocheirsinensis)肝脏中超氧化物歧化酶(Superoxidedismutase,SOD)、过氧化氢酶(Catalase,CAT)、谷胱甘肽过氧化物酶(Glutathioneperoxidase,GPx)活性变化的影响。结果表明,与镉中毒对照组相比,添加VC组能延缓抗氧化酶活性降低时间,减小其降低幅度。VC添加量增加(0.5%上升到1%),抗氧化酶降低的延缓减小作用随之增加。而投喂添加了VE的饲料,河蟹肝胰腺中SOD,CAT和GPx的变化与投喂VC组相似,表现为SOD、CAT基础活性下降,GPx活性上升,同时也能减慢抗氧化酶活性降低时间,减小其降低幅度。随VE量增加(0.02%增加到0.06%)以上变化呈增强趋势。从实验结果看VC,VE都可在镉致毒初期起到抗氧化保护作用。     

红海湾远海梭子蟹Portunus pelagicus的食物组成及营养位置分析

利用稳定同位素技术;分析了2015年5月在红海湾海域采集的远海梭子蟹Portunus pelagicus及其潜在食物的碳（δ13C）和氮（δ15N）稳定同位素比值;通过稳定同位素混合模型（SIAR）;估算不同食物对远海梭子蟹食性的相对贡献。结果表明:远海梭子蟹的δ13C值在-18.1‰~-14.1‰之间;δ15N值的范围为9.8‰~13.8‰;平均值分别为-16.9‰±1.3‰和12.1‰±2.3‰。δ13C和δ15N值的变化范围均较大;表明远海梭子蟹食物来源广泛、多样。远海梭子蟹的食物主要由双壳贝类、螺类、鱼类、头足类、虾类、蟹类和藻类组成。SIAR模型计算结果显示:藻类为远海梭子蟹的主要食物;其平均贡献为32.5%;双壳贝类的相对食性贡献为21.9%;虾类、螺类、鱼类和蟹类的平均贡献差异较小;分别为9.7%、9.7%、9.6%和9.3%;头足类的贡献最低;仅为7.3%。此外;根据δ15N值及营养位置的计算公式得出:远海梭子蟹在红海湾海域位于2.91±0.68营养级。远海梭子蟹的7类潜在食物中;藻类的营养位置最低;为1.21±0.58;其次为双壳贝类;为2.00±0.25;蟹类、螺类、虾类和鱼类的营养位置分别为2.85±0.33、2.87±0.26、3.01±0.16和3.08±0.18;头足类的最高;为3.41±0.17。     

Gill fouling and parasitism in the rock crab, Cancer irroratus say

Rock crabs, Cancer irroratus, were examined grossly and microscopically to estimate the influence of molting activity, geographical source and seabottom sediments on gill color, carapace color and tissue pathology. Collections were made from deepwater stations (20–55 m) in the Hudson Shelf Valley near dredge- and sewage-sludge disposal sites in the New York Bight apex, and from nearshore waters near Boothbay Harbor, Maine. Extensive gill blackening was noted in up to 30 % of the crabs from the New York stations when all specimens were in the intermolt condition. Blackening was not noted in intermolt crabs collected in Maine. Carapace blackening was present in up to 79 % of the New York crabs but absent in those from Maine. Severe blackening was attributed to direct contact of Cancer irroratus with blackened, highly organic sediments in the Hudson Shelf Valley. Fouling microorganisms on gill epicuticle included bacteria, diatoms, stalked ciliates, amoebae, flagellates and copepods. Bacteria, diatoms, amoebae and flagellates were common on crab gills from both locations. Ciliates and copepods counted in histological sections were considerably fewer in crabs from the New York stations than in those from Maine. Melanized nodules (hemocyte clumps) were present in gill tissue in up to 58 % of the New York crabs and in up to 68 % of Maine crabs. Unidentified microsporidans were found in up to 55 % of the New York specimens and in less than 2 % of those from Maine. Histological data accumulated since 1975 are summarized to provide a list of endoparasites, polychaete worms and barnacles observed in Cancer irroratus from the northeastern United States. Gill blackening, but not parasitism or microbial fouling, proved to be the best indicator or monitoring the effects of ocean dumping on the health of a benthic crustacean species.     

日本蟳捕食栉孔扇贝机制的初步研究

在现场和室内条件下研究了日本蟳(Charybdis japonica)对栉孔扇贝(Chlamys farreri)的捕食机制。现场实验表明,成年日本蟳可以捕食壳高小于5.0 cm的栉孔扇贝,其捕食强度随着水温的升高而增大,而壳高大于5.9 cm的栉孔扇贝则可以免遭日本蟳的捕食;相对于栉孔扇贝,日本蟳更倾向于捕食贻贝;室内实验表明水温低于10℃时,日本蟳对大规格扇贝的捕食作用不明显。相同温度条件下,室内实验日本蟳的捕食强度要低于现场,但其温度系数(Q10)差别不大。在上述实验基础上,本文提出了提高底播栉孔扇贝成活率的方法,即选择大规格的扇贝在水温较低的秋、冬季进行底播,同时添加一部分贻贝作为牺牲诱饵以减少日本蟳对扇贝的捕食。     

锯缘青蟹大颚器的显微和超微结构

采用组织切片和电镜技术,对锯缘青蟹大颚器的形态结构进行了观察.大颚器位于大颚肌几丁质腱的后侧基部.腺细胞胞质丰盈,质膜大量内陷和作不同程度的卷绕,内含大量具管状脊的线粒体和滑面内质网.卵巢成熟时大颚器腺细胞内线粒体增多,少数线粒体形态异样,高尔基体易见,散布的小管状滑面内质网(tSER)少,高度汇合的tSER增多,大小形状不同的空泡数量明显增加,且近质膜分布,质膜内陷和卷绕程度加剧.表明此时锯缘青蟹大颚器的合成与分泌能力增强,其生理功能与卵巢发育密切相关.     

Effects of long-term exposure to silver or copper on growth, bioaccumulation and histopathology in the blue mussel Mytilus edulis

The purpose of this study was to determine the long-term accumulation of either silver or copper from low concentrations in seawater by blue mussels, Mytilus edulis. Mussels raised from eggs in the laboratory to the age of 2·5 months (approximately 4·5 mm in length) were continuously exposed to 0, 1, 5 and 10μg/liter of either silver (nitrate) or copper (chloride) and sampled at 12, 18 and 21 months for growth studies, measurements of metal accumulation and histopathological examination.Whole-body soft tissues were analyzed for the presence of both silver and copper, as background levels of copper in the incoming seawater averaged 2–4 μg/liter. Mussels exposed to silver had accumulated significant amounts of silver only at the highest test concentration (10 μg/liter Ag) after 12 months, but at 18 and 21 months significant levels were accumulated at all three test concentrations. Mussels exposed to copper accumulated significant amounts of copper at 5 and 10 μg/liter Cu after all three sampling periods, but not at 1μg/liter. Silver-exposed animals also accumulated significantly greater amounts of copper than control animals.In a comparative study, field-collected juvenile mussels (approximately 16·1 mm in shell length) and adult mussels (approximately 53·4 mm in shell length) were exposed for 12 months to 0, 5, 25 and 50 μg/liter silver only and subsequently sampled for metal-accumulation analyses and growth measurements. Juvenile mussels accumulated significant amounts of silver at all test concentrations, with the exception of mussels exposed to 5 μg/liter Ag for 6 months. Copper accumulation in the silver-exposed juveniles was significant only at 50 μg/liter Ag after 6 months, but at all test concentrations after 12 months. Adult mussels exposed to silver accumulated significant levels of both silver and copper, but at somewhat lower levels than juveniles.In the growth study, silver had no effect on laboratory reared mussels at the highest concentration of 10 μg/liter tested, whereas copper at 10 μg/liter did appear to affect growth as early as 4 months after the start of experimental exposure. Field-collected juvenile mussels did show inhibition in growth after 6 months' exposure to 25 and 50 μg/liter Ag, with some growth occurring after 12 months. Adults also showed inhibition in growth after 6 months but not at 12 months.Histopathological examination of mussels exposed to either 5 or 10 μg/liter of copper for 18 months showed changes in the digestive diverticula, gastrointestinal tract, reproductive tract and muscle tissues. These changes were more noticeable in mussels exposed to 5 μg/liter Cu than in those exposed to 10 μg/liter. Mussels exposed to silver for 21 months showed yellowish to black particulate deposition in the basement membrane and connective tissue of the various organs and tissues. Silver deposition increased with increasing test concentration.     

Lunar rhythms in the egg hatching of the subtidal crustacean: Callinectes arcuatus Ordway (Decapoda: Brachyura)

The Gulf of Nicoya, an estuary on the Pacific coast of Central America, contains a large population of the portunid crab Callinectes arcuatus. Results of a 12 month survey indicated that spawning activity occurs throughout the year, but with a distinct peak during the five-month dry season (December-April). Mature females were most prevalent in the upper regions of the gulf during the rainy season and appeared to migrate to the lower gulf to spawn during the dry season. Patterns of spawning and apparent migration differed from those reported in an earlier study of C. arcuatus along the Pacific coast of Mexico.     

泥蚶初级卵母细胞发育及卵黄发生的超微结构研究

利用透射电镜技术观察了泥蚶(Tegillarca granosa)初级卵母细胞发育期间各细胞器的特征及卵黄粒的形成过程.结果显示:泥蚶初级卵母细胞的发育过程可以分为卵黄合成前期、卵黄合成期及卵黄合成后期三个时期.在卵黄发生过程中,线粒体、高尔基体、粗面内质网、微吞饮泡等都参与了卵黄粒的形成,且线粒体和粗面内质网是形成卵黄粒的主要胞器.卵黄发生的原料以外源性合成为主,外源性营养物质主要来自消化腺.     

Effect of crab size and habitat type on the locomotory activity of juvenile shore crabs, Carcinus maenas

Post-settlement processes are a major focus in the study of the dynamics of marine populations and communities. Post-settlement movement of juveniles is an important, but often ignored, process which affects local predator–prey and competitive interactions. We used benthic suction sampling and pitfall traps to examine density and locomotory activity of Carcinus maenas juveniles in different intertidal habitat types in the Rio Mira Estuary, Portugal, to better understand intra-specific interactions in a system where density-dependent processes are known to regulate population dynamics. As expected, significantly higher densities of juvenile shore crabs were found from bare mud compared to densely vegetated habitats. At the time of sampling, small and intermediate stages together outnumbered by far the larger juveniles. Conversely, larger crabs were much more frequent than smaller ones in traps. A locomotory index (LI), i.e. the ratio between crab catch in pitfall traps and their density within their moving range, is proposed as a measure of movement. LI analyses indicated that: (1) movement is an order of magnitude higher in large than small juveniles and much higher in sparse than dense vegetation cover; (2) activity of small juveniles is mostly crepuscular, regardless of vegetation cover; and (3) movement of large juveniles is very limited in dense Zostera patches, but very high in sparsely vegetated areas, during the day and night. These results suggest that small juveniles are relatively protected under dense vegetation cover due to lower mobility of larger crabs, and provide evidence of temporal segregation of activity windows between juvenile crabs of different sizes, which may be a key mechanism to reduce cannibalism and therefore increase the carrying capacity of nursery habitats.