Identification of transcriptional effects of ethynyl oestradiol in male plaice (Pleuronectes platessa) by suppression subtractive hybridisation and a nylon macroarray

Suppression subtractive hybridisation (SSH) was used to generate cDNA libraries representing genes differentially expressed in response to ethynyl oestradiol (EE2) exposure in liver from male plaice (Pleuronectes platessa) previously analysed for vitellogenin (VTG) induction. Characterisation of the cDNA clones identified many as VTG (2 genes) and zona radiata proteins (ZRP) (3 genes), but 40 encoded other proteins, with more than half cryptic. Further analysis identified 85 non-redundant clones suitable for array on nylon membrane. Radiolabelled cDNAs were prepared from hepatic mRNA from EE2 treated plaice (0 and 21 days) and hybridised with the arrayed clones. Analysis of the data showed that 11/17 novel, 21/22 VTG, 13/14 ZRP, 2/2 liver aspartic proteinase (LAP) and 8/10 other mRNAs were up-regulated by EE2 exposure.     

The distribution and population dynamics of O-group plaice (Pleuronectes platessa L.) on nursery grounds in the Firth of Forth

Midsummer (1 August) population estimates of about 2 million O-group plaice (Pleuronectes platessa L.) were derived for sandy bays around the Firth of Forth in 1979–1980. This is an order of magnitude less than similar estimates made for the Clyde Sea Area in 1973–1974. Autumn population estimates of 0·4–1·0 million fish were comparable to estimates by the Ministry of Agriculture, Fisheries and Food for the area between the Scottish border and Flamborough Head (2·3 million for 1970 and 1973) which represented 4·8% (1973) to 5·3% (1970) of the total number of O-group fish on the English east coast.Largo Bay was the most important nursery area holding 25% of the total population. It is particularly well situated to receive newly metamorphosed plaice carried in water currents along the north side of the Forth from the spawning ground off Fife Ness. Plaice in the Forth are mainly distributed on fine to medium sandy beaches (186–480 μm), the mean number per haul in midsummer (D) being correlated with the median diameter (m.d. in μm) of the low water sediments by the equation: D=−45·7666+0·2327 m.d. (n=11,r=0·68,P<0·02 but>0·01).The shallow inshore water in sandy bays in the outer Firth was well mixed and more marine than estuarine (27·7–35·0‰). The correlation coefficient between fish density and water temperature was low, while that with salinity (S‰) was: D=6·1618+0·2238S (n=23,r=0·62,P<0·005).Regression analysis demonstrated that the relationship between the instantaneous mortality rate (Z) and the initial population density (D_p) was: Z×100=0·7480+0·0546d_p (n=12,r=0·87,P<0·001).The mean mortality rate for the O-group plaice in the Forth nursery areas was 53% month⁻¹.     

Development of habitat models for Nephtys species (Polychaeta: Nephtyidae) in the German Bight (North Sea)

The aim of the study was the development of habitat models for Nephtys species (Polychaeta: Nephtyidae). The investigation area was the German Bight, the southeastern part of the North Sea. Models were developed based on field data collected between 2000 and 2006. In addition, data on environmental variables were retrieved from long-term monitoring data sets and from the sediment map by Figge [Figge, K., 1981. Nordsee. Sedimentverteilung in der Deutschen Bucht. Map No. 2900. Publisher: Deutsches Hydrographisches Institut, Hamburg]. The statistical modelling technique used was multivariate adaptive regression splines (MARS). Models were fitted individually for each species. Evaluation of predictive discrimination and predictive accuracy of the developed models was by calculation of the area under the receiver operating curve (AUC) or sensitivity and specificity, respectively. Habitat models with best predictive fit were selected for the presentation of habitat suitability maps.Six Nepthys species were found: Nephtys assimilis, N. caeca, N. cirrosa, N. hombergii, N. incisa and N. longosetosa. N. hombergii was most common whereas N. incisa and N. longosetosa were rare. Habitat preferences varied considerably among the species. For all investigated Nephtys species except N. longosetosa a habitat model could be developed based on four predictor variables. The habitat models with best predictive fit were those for N. cirrosa and N. hombergii. The N. caeca habitat model was of limited predictive accuracy and only accept predictive discrimination. The number of predictors as well as the relative importance of the respective predictors in the model varied among the different species. Direct comparison of most suitable habitats for the different species based on modelling revealed that in the mostly sandy regions parallel to the German coast in water depths up to 20 m an overlap between N. caeca, N. hombergii and N. cirrosa exists. In the deeper central German Bight with mostly fine sands with increased mud contents N. hombergii, N. assimilis and, at least partially and rare in numbers, N. incisa co-occur. It can be concluded that important sediment characteristics like grain size median and mud content as well as water depth and mean salinity are useful parameters to describe the habitat requirements of most Nepthys species in the German Bight. However, additional variables need to be incorporated into such analyses.     

The effect of salinity on growth and weight loss of juvenile plaice (Pleuronectes platessa, L): An experimental test

Previous population estimates of the 0+ plaice (Pleuronectes platessa L.) in the Firth of Forth, east central Scotland, did not take account of the Forth estuary west of the Forth bridges. Previous work found plaice in the estuary grew as fast as, or faster than, the outer firth plaice. It was hypothesised that salinity may affect growth rates of early 0+ plaice. This hypothesis was tested in a laboratory experiment, by exposing juvenile plaice to three different, but naturally — experienced by the juveniles, salinities; 25, 30 and 35. Plaice fed a minimum ration did not grow in length. Mean weight decreased at all three salinities, however, the lowest weight loss was found at the lowest salinity (25) and the highest weight loss was found at the highest salinity (35). The minimum feeding ration was halted and plaice were then fed ad libitum. Consumption rates were not significantly different during the ad libitum feeding, while significant differences in mean weight change were found between the highest and lowest salinities.     

Rearing in intermediate salinity enhances immunity and disease-resistance of turbot (Scophthalmus maximus L.)

Studies were conducted to investigate the non-specific immune responses and disease-resistance of juvenile turbot Scophthalmus maximus,cultured at four different salinities(8,20,32 and 40) . Three concentrations(3.75 × 10 7,3.75 × 10 8 and 3.75 × 10 9 CFU/ml) of Vibrio anguillarum suspension were employed at each salinity to determine the 4-day LD 50 . The serum lysozyme activity,the alternative complement pathway activity(ACH50) and the phagocytosis percentage of head kidney in turbot were tested at 24,48 and 72 h post-challenge of V. anguillarum(1.1 × 10 8 CFU/ml,0.1 ml) ,respectively,to evaluate the non-specific immune responses at the selected rearing salinities. Fish reared at salinity 20 had the lowest mortality,namely,the highest 4-day LD 50 value(8.88 ± 0.17) . Besides,the lysozyme activity,ACH50 and the phagocytosis of turbot were the highest at the salinity 20,but with the lowest at the salinity 40 treatment regardless of sampling time. In addition,the non-specific immune activities kept increasing within 72 h post-challenge of V. anguillarum,except that the lysozyme activity increased from 24 to 48 h,and then decreased from 48 to 72 h at 40 significantly. These results together indicate that rearing in intermediate salinity(20) was able to enhance the immunity and disease-resistance of turbot.     

Demersal fishing disturbance increases predation risk for whelks (Buccinum undatum L.)

Field observations by divers indicated that a high rate of predation of whelks (Buccinum undatum) by starfish (Asterias rubens) occurred in an area disturbed by scallop dredging, although these whelks mostly appeared to be alive and externally undamaged. The ability of whelks to escape from starfish was tested in the laboratory after they were dropped or rolled to simulate direct physical contact with bottom fishing gear. Dropping whelks did not significantly affect their escape behaviour, but whelks which had been rolled took significantly longer to right themselves and were significantly less likely to perform an escape response than whelks that had not experienced this treatment. This study suggests that demersal fishing may indirectly increase whelk mortality by increasing their risk of predation.     

Using oocyte size to assess seasonal ovarian development in Solea solea (L.)

Maximum oocyte size was used to assess seasonal ovarian development in sole. Fish age, especially the adolescent period, appeared to affect the start of vitellogenin-dependent oocyte development in the annual reproductive cycle and the subsequent oocyte growth rate. The majority of oocyte growth occurred between September and March. Several other aspects of ovarian development were also age-dependent, including the increase in ovary condition factor (ovary weight/fish length³) and the size of oocytes commencing nuclear migration. Evidence is presented that in the recruiting year class of sole abortive maturation occurs where oocytes develop yolk but spawning does not take place. The implications of this study on the estimation of female spawning stock biomass are discussed.     

Factors involved in growth plasticity of cockles Cerastoderma edule (L.), identified by field survey and transplant experiments

In Arcachon Bay, a macrotidal coastal lagoon in southwest France, growth rates of cockles Cerastoderma edule (L.) differ greatly between locations. Sampling of populations at different tidal levels showed that the mean shell length was significantly and positively correlated with immersion time, whereas no correlation was found with population density and microphytobenthos biomass. Transplants of cockles between two intertidal sites were used to examine the relative importance of habitat and site of origin for growth rate and condition index. Artifacts due to manipulation were assessed, i.e. the impact of enclosures on growth, conditon index and mortality. During a 5-mo reciprocal transplant experiment, growth rates of the transplanted cockles and the cockles already present were similar, whereas the sites of origin did not affect growth rate. Condition index, however, displayed significant differences in relation to both transplant and origin sites. During a further 6-mo experiment in which cockles were transplanted from one site to four new sites, growth was mainly influenced by tidal level. These results indicate that net growth started when cockles were immersed 30% of the time. It is concluded that, on the scale of Arcachon Bay (156 km²), differences in growth and condition between cockle populations are largely phenotypic. Tidal level (i.e. food supply and emersion stress) accounts for most of these differences, but the existence of genetically or physiologically different populations cannot be excluded.     

Population dynamics of the venerid bivalve Callista chione (L.) in a coastal area of the eastern Mediterranean

The venerid bivalve Callista chione is the most prominent suspension-feeding bivalve inhabiting the soft bottom of coastal areas in the northern Euboikos Gulf (Aegean Sea), where it is exploited by small-scale fishery. The parameters of the von Bertalanffy growth function were estimated and the somatic production-to-biomass ratio was found to be 0.45 y⁻¹. The maximum age of adult individuals was 17 years. The species is gonochoristic, its oocyte diameter ranged from 54.5 to 120 μm with a conspicuous jelly-coat. Microscopical examination of the gonads showed that gonadal production did not start until the second year of life. The observations on the gonadal cycle indicate that this bivalve is capable of year-round reproduction.     

Vertical migrations of the mangrove snail Cerithidea decollata (L.) (Potamididae) through a synodic month

The vertical migration on mangrove trunks of the gastropod Cerithidea decollata was followed for 5 weeks, in a Kenyan mangrove. Most of the times, snails forage on the mud surface, during low tide, and climb back on trees well before the incoming tide. As soon as the sea retreats, the downward migration takes place and the snails spread again on the ground. The migratory behaviour of snails can vary widely, depending on the relative tide intensity, and different strategies can be exhibited. Individuals can spend several days on trees without migrating to the ground, around Spring Tides, or else, they might remain on the ground without bothering to migrate upwards, during Neap Tides, when the study area is not reached by the water. These irregular animal behaviours, relating to the complicated tide succession, can hardly be explained by the sole presence of an internal clock, and direct cues seem necessary to switch between different strategies, tuning the snails migratory behaviour to the actual local sea conditions.     

Food and feeding habits of juvenile flounder Platichthys flesus (L.), abd turbot Scophthalmus maximus L. in the åland archipelago, northern Baltic Sea

The food choice of juvenile flounder (Platichthys flesus) and turbot (Scophthalmus maximus) was studied in the northern Baltic Sea during the years 1988, 1989, 1994 and 1995. The diet included organisms from 30 species/taxa in flounder (n = 306) and 10 species/taxa in turbot (n = 41). Flounder 45 mm mainly consumed meiofauna (dominating taxon: Harpacticoida, Copepoda) and larger fish (46–101 mm) consumed macrofauna (dominating taxa: Oligochaeta, Amphipoda and Chironomidae). In terms of biomass, macrofauna dominated for all sizes of flounders, and meiofauna was important only for the smallest fish. A strong seasonal variation could be detected in the diet. In spring, macrofauna dominated for all size classes of fish (only fish > 30 mm were caught in spring), while in summer and autumn meiofauna dominated the diets for fish 45 mm in size. Juvenile turbot (22–88 mm) consumed macrofauna and small fish. Turbot 30 mm consumed mainly amphipods, while > 30 mm turbot consumed mysid shrimps, amphipods and fish.The ontogenetic shift from meio- to macrofauna-sized prey in flounders occurs at a larger fish size in the northern Baltic Sea than reported in other areas, possibly depending on the increased relative importance of meiofauna in the northern Baltic. The seasonal variation in the diet could be due to seasonally changing abundances in the zoobenthos, or for the small fish (1-group, spring), to switching from meio- to macrofauna in order to optimize their energy gain. The 0-group flounders consumed meiofauna for a long period, possibly due to a learning-process or simply due to easy availability of meiofauna. Turbot has a much larger mouth gap than flounders, thus allowing them to consume macrofauna from the beginning of their benthic life.     

Nursery areas of red mullet (Mullus barbatus), hake (Merluccius merluccius) and deep-water rose shrimp (Parapenaeus longirostris) in the Eastern-Central Mediterranean Sea

The spatial pattern of the nursery areas of red mullet (Mullus barbatus), hake (Merluccius merluccius) (Linnaeus, 1758) and deep-water rose shrimp (Parapenaeus longirostris) (Lucas, 1846) was studied in the South Adriatic and North Ionian Seas (Eastern-Central Mediterranean) applying geostatistical techniques and data from time series trawl surveys conducted in the area. The analysed variables were: R (number of recruits/km²) and R/Tot (fraction of recruits on the total sampled population). The structural analysis showed a spatial pattern of both variables characterized by continuity on a small scale. Predictions of nursery area localization with probability of finding recruits at different threshold values were obtained through median indicator kriging. For the red mullet the nurseries were mainly identified in the South Adriatic Sea off the Gargano peninsula and between Molfetta and Monopoli within 50 m in depth. The main concentration of hake juveniles was found to be between 100 and 200 m in depth along the Gargano peninsula and between Otranto and Santa Maria di Leuca, where a nursery of deep-water rose shrimp was also detected. An overlapping depth, between 100 and 200 m, was identified for hake and deep-water rose shrimp nurseries. Protection of these areas through limitations of fishing pressure is discussed.     

大黄鱼cGAS-like基因isofrom X1的克隆鉴定及在感染刺激隐核虫后的表达变化

cGAS(cyclic GMP-AMP synthase)基因是近几年来新发现的先天免疫中一个新型的信号分子。作者基于大黄鱼(Larimichthys crocea)基因组信息,用cDNA末端快速扩增(RACE)技术在鳃组织中克隆鉴定了cGAS基因cDNA全长序列:包含1个231 bp的5′端-非翻译区(5′-UTR)、207 bp的3′-UTR和1 488 bp的开放阅读框(ORF),命名为Lc-cGAS-like X1。该基因编码1个由495个氨基酸残基组成的多肽,预测的分子量(Mw)大小为56.57kDa,理论等电点(pI)为9.45;序列分析表明其预测的蛋白无信号肽,第115～431位氨基酸序列为Mab-21功能域,第470～492位氨基酸序列为跨膜结构域。基因组全长3 153 bp,包含8个外显子和7个内含子,所有内含子的剪切特点都符合GT-AG规则。同源比对结果显示:Lc-cGAS-likeX1与鱼类cGAS的相似性大于66%,与高等脊椎动物的相似性较低。系统进化分析表明Lc-cGAS-like X1与鱼类cGAS聚成一支。荧光定量PCR(qPCR)检测显示Lc-cGAS基因(X1/X2两种剪切体)在大黄鱼9种组织中均有表达,其中在鳃中的表达量最高;刺激隐核虫感染后,大黄鱼免疫器官头肾中Lc-cGASmRNA分别在刺激隐核虫幼虫感染阶段和滋养体脱落阶段出现了极显著上调(P0.01);解毒器官肝脏中分别在滋养体脱落阶段和细菌感染阶段出现极显著上调变化(P0.01)。本研究结果暗示大黄鱼cGAS基因参与了大黄鱼感染刺激隐核虫的免疫防御过程,为进一步了解鱼类cGAS基因的多样化功能提供参考。     

溶藻弧菌对三疣梭子蟹抗氧化酶系统的影响

从患乳化病的养殖三疣梭子蟹(Portunus trituberculatus)体组织内分离出痛原菌--溶藻弧菌(Vibrioalginglyticus),通过肌肉注射的方式对三疣梭子蟹进行感染实验,对照组注射等量的生理盐水,在感染后O、24、48和72 h分别采集不同处理组三疣梭子蟹的血淋巴、肌肉和肝胰腺组织,测定溶藻弧菌对其各组织抗氧化相关酶活的影响.结果表明:注射生理盐水的对照组三疣梭子蟹体组织中抗氧化酶和MDA含量在不同的采样时间变化不显著(P>0.05).与对照组比较,三疣梭子蟹感染溶藻弧菌后血淋巴、肌肉和肝胰腺组织中SOD活性随着感染时间的延长显著降低(P<0.05).在感染溶藻弧菌24 h时血淋巴和肝胰腺组织中GSH-px活性有增强的趋势(P>0.05),随后在感染48 h后GSH-px活性显著降低(P<0.05);感染组各体组织中的MDA含量均随着感染时间的延长而显著上升(P<0.05).结果显示梭子蟹感染溶藻弧菌后机体消除氧化过程自由基的抗氧化酶活降低,脂质过氧化物水平升高,使得抗氧化防御能力下降.     

基于耳石中锶和钙元素分析破解日本六角川和筑后川河口水域刀鲚的生境履历

为了解日本有明海沿岸六角川和筑后川河口水域鳀科鲚属鱼类刀鲚（Coilia nasus）的洄游特征和生境履历,利用电子探针微区分析（EPMA）技术,对采于两水域刀鲚的矢耳石进行了锶（Sr）元素含量的X射线强度面分布及耳石最长轴锶钙元素比值（Sr:Ca）的定量线分析。结果发现10尾六角川刀鲚（LJC）中的7尾以及15尾筑后川刀鲚（ZHC）耳石核心区域的Sr:Ca比值≤ 3,显示出这些个体在淡水生境出生并早期生活于淡水的履历。紧邻核心区域随后的生长阶段,Sr面分布图谱显示出了多样对应于含量水平差异的不同颜色同心环轮变化样式,Sr:Ca比值的波动亦可相应地分成1-6个差异极其显著的阶段,显示出这些个体在随后生活史阶段中极富变化的生境转换状况。另外3尾六角川刀鲚耳石核心的Sr:Ca比值较高（3-6.7）,对应于半咸水生境;但在核心区域之外,Sr:Ca比值则高低变化不一,显示出这些六角川刀鲚个体在河口半咸水生境孵化的特征。本研究表明,二维Sr元素含量面分布图谱和基于耳石最长轴的Sr:Ca比值完全可以作为重建日本有明海沿岸各河口水域刀鲚生境履历的有效工具。     

天鹅湖大叶藻种苗补充情况调查

本研究以山东荣成典型潟湖——天鹅湖内的大叶藻(Zostera marina L.)海草床为研究对象,从2015年3月到6月对大叶藻种子萌发形成的幼苗进行进了月度调查,包括春季大叶藻种苗的密度、株高、叶鞘高、叶数、叶宽以及分枝情况。结果表明,天鹅湖大叶藻种子在3月即开始萌发,但萌发量较少,种苗密度较低;4月种子大量萌发,种苗密度不断升高,并达到峰值,为(481.77?303.42)株/m2;5月份种子萌发结束,种苗密度下降,同时开始克隆生长;6月份种苗不断生长,分枝数增多,种苗开始出现有性生殖枝。至此,春季种苗的萌发过程结束,进入有性生殖过程。     

连续降温对大菱鲆成鱼代谢机能的影响

为了探寻连续降温对大菱鲆(Scophthalmus maximus)成鱼血清、肝脏及肌肉能量代谢等指标的影响,作者将大菱鲆从正常养殖水温18℃快速、连续降温至1℃,并在18℃、13℃、8℃、5℃、3℃和1℃共计6个温度点取血采样,分别测定血清中的总蛋白(TP)、白蛋白(ALB)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、甘油三酯(TG)、总胆固醇(TC)、血糖(GLU)以及肝脏中的糖原和肌肉中的乳酸等指标。结果发现,随着温度降低,大菱鲆的血清TP浓度呈先下降后升高的趋势,各实验组与对照组无显著差异(P0.05),但在13℃和3℃之间出现显著性差异(P0.05);血清ALB浓度则呈现升高趋势,在3℃时显著高于1℃(P0.05);血清TC浓度呈现升高趋势,且3℃时显著高于18℃(P0.05);GLU浓度呈先降低后升高趋势,且8℃、5℃、3℃实验组浓度显著高于其余实验组(P0.05);尿素浓度呈先降低后升高再降低趋势,且8℃时显著高于1℃(P0.05);肝糖原浓度呈先降低后升高趋势,且8、5、3℃实验组浓度显著高于其余实验组(P0.05);肌肉中乳酸含量呈先升高后降低趋势,且5℃时显著高于18℃(P0.05);其余指标则无显著性差异(P0.05)。结果表明:当温度在18~8℃时,降温对大菱鲆体内代谢影响不大;当温度降到8~3℃时,大菱鲆体内开始出现应激反应;当温度继续降低至1℃时,大菱鲆体内的各种代谢基本处于停滞状态。综合分析各温度下的代谢指标得出结论,3~1℃温度区间与大菱鲆的生态冰温点比较接近,可以在此温度区间附近进一步进行无水保活的探索,在保证大菱鲆无水运输可行性的基础上优化经济成本。     

Contrasting shell morphology, ingestion and grazing preferences in the neritid gastropod Smaragdia viridis (L.) on two seagrass species

The neritid Smaragdia viridis represents the only known native marine mollusc that feeds on seagrass tissues in the European coasts, displaying a strong association with the seagrasses Cymodocea nodosa and Zostera marina in southern Spain. Seasonal dynamics, shell and radular morphology, growth and feeding of this gastropod have been studied in relation to each seagrass species for contrasting trends resulting from a different type of substrate and food source. In both seagrass species, stable populations of this gastropod occur at similar densities and displaying similar growth rates. Nevertheless shells of individuals from C. nodosa are narrower than those from Z. marina and some differences, possibly a consequence of increased wearing on C. nodosa, were noted amongst the radulae. In C. nodosa, a pre-ingestive selection for young epidermal tissues occurs as it was previously observed in Z. marina. The ingestion rate is higher in C. nodosa than in Z. marina but the absorption of ingested tissues is lower in the former. If both seagrasses are present, most individuals ingested preferentially Z. marina rather than C. nodosa, probably due to the lower digestibility of the epidermal tissues in the latter. Seagrass beds, especially those of Z. marina, are suffering a strong regression in southern Spain and the presence of stable populations of this neritid may be restricted to other declining seagrass species in the area.     

针胞藻海洋卡盾藻(Chattonella marina)和硅藻中肋骨条藻(Skeletonema costatum)藻际细菌的分离、鉴定及影响研究

对实验室培养的两株海洋微藻海洋卡盾藻和中肋骨条藻的可培养细菌进行了分离鉴定,分别在海洋卡盾藻和硅藻中肋骨条藻不同生长时期分离得到48株和34株可培养细菌,去除重复序列后得到12株不同的菌株。这些菌株分属α-变形杆菌纲、γ-变形杆菌纲、拟杆菌和放线菌。α-变形杆菌纲在两种藻的藻际环境中占据优势,其中红杆菌科是最为常见的科,中肋骨条藻的藻际细菌比海洋卡盾藻更为多样化。海洋卡盾藻生长后期细菌密度大幅度上升,可能导致了藻细胞的衰亡。中肋骨条藻相关细菌中有一株对旋链角毛藻的生长具有明显抑制作用,而大部分海洋卡盾藻相关细菌对角毛藻的生长具有抑制作用。本研究结果说明海洋卡盾藻的藻际细菌也许对其与硅藻的种间竞争中具有一定作用。