中间球海胆乳酸脱氢酶基因克隆及其对海水酸化的响应

为明确中间球海胆乳酸脱氢酶(LDH)基因序列及表达特征,研究其对海水酸化胁迫的响应,实验利用cDNA末端快速扩增(RACE)技术获得中间球海胆LDH基因(命名为SiLDH)的全长cDNA,并且比较了海水酸化胁迫下,中间球海胆不同组织SiLDH基因表达及总LDH活性的变化情况。结果显示:①SiLDH基因的cDNA全长为1 499 bp,包含133 bp的5′非编码区,349 bp的3′非编码区和1 017 bp编码338个氨基酸的开放阅读框(ORF)。SiLDH编码蛋白的相对分子量为36.40 ku,理论等电点为6.55,属于无信号肽的非跨膜、温和疏水蛋白。②生物信息学分析显示,SiLDH蛋白序列与紫球海胆LDH X4蛋白序列一致性最高(90.88%)。③实时荧光定量PCR(qRT-PCR)发现,SiLDH基因在检测的4种组织中均有表达,相对表达量从高到低为性腺管足肠体腔液;总LDH活性检测显示,中间球海胆总LDH活性从高到低为性腺管足体腔液肠;④海水酸化处理60 d后发现,与自然海水组(pH 8.06±0.01)相比,SiLDH基因在中间球海胆性腺、管足和肠道中呈现总体降低趋势,在体腔液中则呈现随海水pH降低先降低后显著升高的趋势;总LDH活性在性腺、管足和体腔液中呈现随海水pH降低而降低的趋势,在肠道中则呈现随海水pH降低先降低后显著升高的趋势。研究表明,中间球海胆面对海水酸化时可能通过调节SiLDH基因的表达和LDH活性来缓解海水酸化带来的负面影响。     

3种饵料模式对中间球海胆生长、性腺产量和性腺品质的影响

研究了3种饵料模式对中间球海胆整个生长周期的生长速度及商品规格性腺产量和品质的影响。选取6月龄中间球海胆,以3种饵料模式海带(E1)、海带+贻贝(E2)、玉米+贻贝(E3)养殖至商品规格,期间每隔2个月对海胆的壳径和体质量进行测量,实验结束时,对海胆的性腺湿重、性腺水分和性腺颜色等进行测量和分析。结果显示:在海胆的各生长阶段,饵料模式对其壳径和体质量均具有极显著影响,3种模式下海胆的壳径和体质量均为E2>E1>E3。饵料模式对海胆大部分生长阶段壳径和体质量的特定生长率具有极显著影响,3种模式下海胆壳径和体质量的特定生长率均为E2≥E1>E3。饵料模式对商品规格海胆性腺产量和性腺品质具有显著或极显著影响,对于性腺湿重,E2模式[雌性:(7.58±1.21)g,雄性:(7.74±1.95)g]表现最优,其次是E1模式[雌性:(4.50±1.20)g,雄性:(4.87±1.02)g],最后为E3模式[雌性:(2.66±1.02)g,雄性:(2.32±0.75)g];对于性腺指数,E2模式(雌性:15.07%±1.84%,雄性:15.83%±3.01%)表现最优,其次为E1模式(雌性:10.49%±2.71%,雄性:11.27%±2.45%)和E3模式(雌性:12.48%±4.33%,雄性:12.78%±3.73%);对于性腺水分和性腺颜色,E2和E1模式表现最优,E3模式表现较差。饵料模式对不同性别海胆的影响差别较小。实验结果表明,单一投喂海带,中间球海胆即可获得较好的壳生长,而添加动物性蛋白可以明显的提高其体质量和性腺产量,海带+贻贝这一模式可应用于提高海胆养殖效率,而玉米对促进海胆的体生长和性腺产量和品质均无有益效果,即使添加动物性蛋白,玉米+贻贝这一模式也很难替代海带有效地养殖中间球海胆。     

A geostatistical approach for the stock assessment of the edible sea urchin, Paracentrotus lividus, in four coastal zones of Southern and West Sardinia (SW Italy, Mediterranean Sea)

Quantitative surveys of the edible sea urchin, Paracentrotus lividus, were conducted in four fishing zones of Sardinia (Southern Italy, Mediterranean Sea), in Autumn 2007. A total of 120 stations were geo-located along a bathymetric gradient ranging from 0 to 10 m. A geostatistical method was used to evaluate spatial patterns in density and to estimate harvestable stocks. Variographic analyses showed that the isotropic Gaussian and spherical models successfully explained the spatial structure of sea urchin assemblages in these areas. Density maps obtained by punctual kriging showed that sea urchin populations tend to be patchy rather than uniform in their density distribution. A combination of mapping and size categories was used to generate diverse scenarios of harvestable stocks (specimens ≥50 mm in diameter) before the start of the current fishing season. We conclude that the geostatistical approach, which takes into consideration the spatial autocorrelation structure of the populations in small areas, seems to be a good estimator of P. lividus density and biomass and for the assessment of its harvestable stocks, and thus provides an initial step towards a scientific approach to the management of local sea urchin fisheries.     

福建沿海试养中间球海胆的初步研究

为明确不同选育群体中间球海胆的遗传多样性和遗传结构,利用SSR-seq技术和15个微卫星位点,对1个家系选育群体 (FP)、1个群体选育群体 (IP)和1个未经选育的普通养殖群体 (CP)的遗传多样性及遗传结构进行了分析。结果显示,15个微卫星位点共检测出112个等位基因,FP、IP、CP 3个群体的平均观测等位基因数 (N_a)分别为5.077、5.133和6.133个,平均有效等位基因 (N_e)分别为2.816、2.873和3.638个,平均观测杂合度 (H_o)分别为0.522、0.441和0.501,平均期望杂合度 (H_e)分别为0.595、0.599和0.667,平均多态性信息含量 (PIC)分别为0.546、0.543和0.623。家系选育群体 (FP) H_e与H_o的差值 (0.073)低于IP (0.158)和CP (0.166),平均固定指数 (F) (0.115)低于IP (0.248)和CP (0.246)。3个群体间遗传分化系数 (F_st)介于0.018~0.176,为中低等程度的遗传分化。分子方差分析 (AMOVA)结果显示,3个群体的遗传变异主要源于个体间。主成分分析 (PCoA)和聚类进化树结果均显示,3个群体之间的亲缘关系较近,其中IP群体遗传分化程度最高。研究表明,3个中间球海胆群体均具有较高的遗传多样性,多代家系选育和群体选育均未明显降低群体的遗传多样性,家系选育更有利于保持群体的杂合度和控制群体的近交水平,群体选育则会提升群体的遗传分化程度。     

Growth and mortality of cultured bay scallops in the Damariscotta River, Maine (USA)

Growth and mortality of hatchery-reared bay scallops, Argopecten irradians, were determined over an 18 month period (July 1994-December 1995) at two locations in the Damariscotta River, Maine, north of its natural distribution. Growth occurred primarily between May and October when mean water temperature exceeded 10 °C. Final mean shell height (56.3 mm) was significantly greater (p 0.0005) at the upriver site than at the downriver site in association with a higher mean water temperature. Cumulative mortality (43-49%) over this period occurred primarily over the winter months, in conjunction with mean water temperatures below 5 °C. Although it is possible to rear bay scallops to commercial size in Maine, the short growing season, high winter mortality, and costs associated with over-wintering and fouling will likely deter commercial culture.     

First report of signs of infection by Coccomyxa-like algae in wild blue mussels,Mytilus spp., in the Gulf of Maine (USA,Maine)

In August 2019, visual inspection of intertidal zones of the Gulf of Maine (ME, USA) revealed young and adult wild blue mussels, Mytilus spp., in Alley Bay (Jonesport area) with the distinctive L-shaped shell deformity (LSSD) and green spots (GS) in the mantle and adductor muscle. LSSD is a characteristic sign of current or previous mussel infection by photosynthetic unicellular alga from the group Coccomyxa, while GS are algal colonies. Based on these findings, this study represents the first report of infection signs by pathogenic Coccomyxa-like algae in mussels from the coastal waters of the Northeastern United States, providing a base for future large scale monitoring of the alga in the region.     

盐度对大连紫海胆生长发育的影响

大连紫海胆是产于西北太平洋沿海水域的一种重要经济海胆,也是我国正在研究发展的新的海水增养殖种类。为搞好其人工育苗与培养殖技术,探讨主要环境因子对其生长发育的影响是必要的。廖玉麟(1982)、廖承义(1987)、今井利为(1986)、伊东义信(1987)、Fuji(1962)等曾就温度和饵料这两个重要环境因子对海胆类的影响进行过研究,研究的对象包括大连紫海胆、马粪海胆Hemicentrotus pulcher-     

Adding value to live, commercial size soft-shell clams (Mya arenaria L.) in Maine, USA: results from repeated, small-scale, field impoundment trials

The soft-shell clam (Mya arenaria L.) fishery in the state of Maine, USA, is worth $5–10 million annually and is primarily based on the sale of live individuals. More than 80% of the catch is sold for the “steamer clam” market that is highly seasonal due to supply and demand. Prices paid to harvesters for live clams throughout the year increases by as much as 70% during a 4-month period between early spring and late summer. If clams harvested in the spring could be held until late summer, a value-added product could be developed in this fishery. From April to August 1996, at an intertidal and subtidal location in eastern Maine, I tested whether it was biologically feasible to impound commercially harvested clams (shell LENGTH=44–75 mm) at densities between 720 and 850/m² using cages and nets (1.12 m²) to protect animals from predators. Survival was 91.7% (n=12) and 91.6% (n=6) at the intertidal and subtidal site, respectively. There was no discernible shell growth during this period and there was no difference between initial and final clam weights. The methodology was transferred to commercial harvesters in two Maine communities: Wiscasset (1997 and 1998) and Perry (2000). Impounded clams ( =21.8 kg/cage; n=18) lost, on average, 5 kg/cage in 1997 and 1998, whereas no significant net loss in wet weight occurred in cages deployed in 2000 ( =23.2 kg/cage; n=3). Differing harvesting and handling methods of individual clammers, prior to impounding clams, likely explains the variation in weight lost during the impounding periods. The difference in price per live kilogram between the beginning of the impounding period and the August sale date resulted in an average gain of $13.60/cage for clammers in 1997 and 1998 and $57.73/cage in 2000. Simple culture techniques can be used to increase the value of the live harvest of soft-shell clams along the coast of Maine. Clam impoundments may be a way for communities that co-manage the public clam resource with the state's marine resource agency to generate funds to pay for traditional management schemes, as well as enhancement programs that employ hatchery-reared juveniles. In addition, an indirect benefit of clam impoundments might be to create spawner sanctuaries since animals are impounded during the time when gamete release occurs.     

Effects of restricted feeding regimes on growth and feed conversion on juvenile green sea urchin,Strongylocentrotus droebachiensis

Groups of juvenile green sea urchin, Strongylocentrotus droebachiensis (average wet weight = 3.3 g), were fed five different dry feed rations (0.2%, 0.4%, 0.8%, 1.2% and 2.4% of their body weight per week) under constant light and temperature conditions for 160 days (Experiment I) in groups to examine growth effects, and for 40 days as individual treatments (Experiment II) to examine feeding efficiency. There was 100% survival of the sea urchins during both experiments. In Experiment I, the lowest ration group (0.2%) had significantly lower growth than the rest of the groups. There was no significant differences in growth between the sea urchin fed ration over 0.4% dry feed of the body wet weight per week. In Experiment II, the lowest feed ration groups (0.2%) had significant lowest growth but had the best feed conversion ratio (FCR), using 0.5 g of feed of dry feed per gram of sea urchin wet weight body growth. The FCR increased with increasing feed ration and the 2.4% group had the poorest FCR, using 1.3 g of feed per gram weight gain. Results from Experiments I and II illustrate that juvenile green sea urchin can grow at a restricted feed ration that is under maximum feed intake, without reduction in growth.     

虾夷马粪海胆溶菌酶基因全长cDNA的克隆与表达分析

本实验采用RT-PCR和cDNA末端快速扩增(RACE)技术克隆得到了虾夷马粪海胆(Strongylocentrotus intermedius)溶菌酶(LYZ)基因的全长cDNA序列。结果表明, 虾夷马粪海胆LYZ基因全长为912 bp, 含有1个480 bp的开放阅读框(ORF), 编码159个氨基酸, 其中第1−20个氨基酸为信号肽, 蛋白计算分子量为17.69 kD, 等电点为7.75。氨基酸比对分析表明, 虾夷马粪海胆LYZ基因与紫球海胆(Strongylocentrotus purpuratus)和刺参(Apostichopus japonicus)的i型LYZ基因相似百分比分别为91.4%和59.3%, 并且含有i型LYZ基因的保守序列DVGSLSCGP (Y)Y(F)QIK, 所以推断本实验克隆的溶菌酶为i型。采用实时定量PCR方法, 以β-actin为内标, 对其在虾夷马粪海胆各组织中的表达进行研究, 发现LYZ基因在围口膜中表达量最高, 其次是齿间肌、管足、肠、体腔液、雄性性腺和雌性性腺。利用脂多糖(LPS)刺激虾夷马粪海胆, 取刺激后不同时间的海胆体腔液, 对该基因的表达差异进行分析。结果表明, 虾夷马粪海胆的LYZ基因在LPS刺激后8 h时表达量最高, 12 h时开始逐步回落, 至36 h时回落至对照组相近水平。本结果可为虾夷马粪海胆免疫学研究及抗病相关分子标记的开发提供参考依据。

     

光棘球海胆的主要卵黄蛋白cDNA序列分析

提取成熟光棘球海胆(Strongylocentrotus nudus)性腺中的RNA做模板,根据NCBI数据库中已知海胆(编号:AB097218、AB192414、AY090112)主要卵黄蛋白MYP cDNA保守序列设计引物,用LA PCR方式分段扩增并测序得到了光棘球海胆MYP cDNA的全序列。扩增的cDNA全长4 061 bp,包含4 047 bp的开放阅读框,共编码1 349个氨基酸。用CluxtalX1.83对光棘球海胆与其他几种已知海胆MYP cDNA及推导的氨基酸序列进行比对,用Mega3.01计算遗传距离及构建进化树,结果表明,光棘球海胆与其他7种海胆的MYP具有高度的同源性。从氨基酸水平上看,光棘球海胆与红海胆(Pseudocentrotus depressus)亲缘关系最近,遗传距离为0.069±0.01;与同科的马粪海胆(Hemicentrotus pulcher-rimus)、紫球海胆(S.purpuratus)、中间球海胆(S.intermedius)的遗传距离分别是0.095±0.012、0.098±0.011和0.101±0.012;与白棘三列海胆(Tripneustes gratilla)、拟球海胆(Paracentrotus lividus)及绿海胆(Lytechinus variega-tus)亲缘关系相对较远,遗传距离分别是0.216±0.017、0.218±0.017和0.535±0.028。获得光棘球海胆MYP cDNA序列可为进一步研究MYP基因的功能和系统的进化分析奠定基础     

Spatial variability in red sea urchin (Strongylocentrotus franciscanus) recruitment in northern California

To better understand the spatial distribution of recruitment in the northern California red sea urchin ( Strongylocentrotus franciscanus ) population, we sampled size distributions at each of 12 locations in 1995 and 1996, two of those locations in 1994, and 5 of those locations in 1997. An index of recent recruitment in these size distributions and an estimate of density of recent recruitment reflect a similar spatial pattern of recruitment. This pattern appears to be determined by the effect of coastal circulation on larval delivery during relaxation of upwelling, and not the result of the positive effect of the adult spine canopy on juvenile survival. Recent recruitment of red sea urchins in northern California is higher in areas more frequently subjected to onshore and poleward flow during relaxation of upwelling. These results are consistent with a mechanism by which alongshore spatial variability in southward, offshore flow during upwelling winds allows larvae to maintain latitudinal position, whereas flows during event-scale relaxations in upwelling winds serve to distribute settling larvae alongshore, favouring areas north of promontories. The consequent spatial pattern of red sea urchin settlement varies from year to year, and there are not yet sufficient data to demonstrate the degree to which this spatial pattern in recruitment determines a spatial pattern in fishable adult abundance.     

Cu2+对中间球海胆生长、免疫及性腺发育的影响

为探讨铜离子(Cu2+)对中间球海胆生长和性腺发育的影响及调控机制,在室内水槽(200 L)中进行为期60 d的浸泡实验。实验设置测试组(0.02 mg/L Cu2+)和对照组(自然海水),每个处理设置3个重复,每个水槽养殖20只海胆(13.58±0.79)g。实验中期和末期分别测定海胆增重率(WGR)、性腺指数(GI)、抗氧化酶活性和主要卵黄蛋白基因(MYP)表达量。结果显示,30 d时,测试组的WGR和GI与对照组差异不显著,而60 d时测试组的WGR和GI显著低于对照组;60 d时,测试组海胆体腔液中过氧化氢酶(CAT)和谷胱甘肽巯基转移酶(GST)活性均显著低于30 d时的对应值,而丙二醛(MDA)含量较30 d时有所升高;60 d时,测试组抗氧化能力总体高于对照组,其中体腔液中CAT活性显著高于对照组。2次取样中,测试组性腺中MYP基因的表达量均显著低于对照组,而测试组消化道中MYP基因的表达量却显著高于对照组。60 d时,测试组海胆体腔细胞中总蛋白含量显著高于对照组,而体腔液上清液中总蛋白含量在测试组和对照组之间差异不显著。研究表明,铜离子能够引起氧化应激,降低海胆的增重率和性腺指数,这可能是通过抑制性腺MYP表达量及阻碍MYP从消化道向性腺正常转运所致。     

Influence of stocking density and temperature on early development and survival of the purple sea urchin,Strongylocentrotus purpuratus (Stimpson, 1857)

We evaluated the effect of four densities (940, 1880, 3760, 7520 eggs cm^?2 and 0.5, 1, 2, 4 ind mL^?1 of embryos and larvae, respectively) and four temperatures (8, 11, 14, 17°C) on early growth and survival of the sea urchin Strongylocentrotus purpuratus. Prism‐stage length was significantly greater in embryos initially held at 940 and 1880 eggs cm^?2 than in those held at 3760 and 7520 eggs cm^?2. Larvae grew significantly faster and had significantly greater survival when reared at 0.5 or 1 ind mL^?1 than when held at 2 or 4 ind mL^?1. Embryos had greater survival at 11 and 14°C than at 8 and 17°C, whereas embryo length was significantly smaller at 8°C than at 11, 14 or 17°C. Larvae grew significantly slower at 8°C than at 11, 14 or 17°C, whereas survival was significantly reduced at 8 and 17°C compared with 11 and 14°C. Per cent survival from prism to metamorphic competency in the best treatments was 48.9 ± 2.2% and 50.0 ± 3.6% (mean ± SE) for the 1 ind mL^?1 and 11°C treatments, respectively. On the basis of these results, for rearing of S. purpuratus under static conditions, we recommend that fertilized eggs and larvae be held at ≤1880 eggs cm^?2 and ≤1 ind mL^?1, respectively, and at 11–14°C.     

Effect of feed deprivation and refeeding on the MYP gene expression of the sea urchin (Strongylocentrotus intermedius)

To determine the main expression site of major yolk protein (MYP) gene and the mechanisms for adaptation to starvation and refeeding in Strongylocentrotus intermedius, MYP mRNA expression amounts were analysed using a real‐time RT‐PCR. The results showed that MYP could be transcribed in the intestine, stomach, gonad and coelomocytes, and that the intestine was the main expression site of MYP gene in non‐starved urchins. The MYP synthesis in the intestine decreased during 15 days of starvation (67.70%, 52.58% and 71.35% of the control at 5, 10 and 15 days of fasting respectively) and then increased dramatically by different amounts (the peaks were 2.71‐, 12.16‐ and 7.89‐fold that of the control respectively) during the refeeding stages. Nevertheless, the expression amounts in the gonads did not decline, but increased continuously during all periods of fasting (2.66‐, 3.72‐ and 13.19‐fold that of the control at 5, 10 and 15 days of starvation respectively) and during the refeeding stages. At the end of the recovery feeding experiment, the levels reached 9.58‐, 17.48‐ and 100.69‐fold that of the control. These data suggested that the ‘priority’ strategy for the sea urchin is to reduce MYP expression amounts in the intestine if food is limited and to increase MYP gene expression in the gonad to protect reproductive function.     

Bluefin tuna (Thunnus thynnus) distribution in relation to sea surface temperature fronts in the Gulf of Maine (1994–96)

Fishery‐linked aerial surveys for bluefin tuna (Thunnus thynnus) were conducted in the Gulf of Maine (GOM) from July through October, 1994–96. Each year, from 507 to 890 surface schools were detected and their locations examined in relation to oceanographic conditions. Correlations between bluefin tuna presence and environmental variables were explored for sea surface temperature (SST), distance to a SST front, frontal density (relative density of all SST fronts seen in a given 1 km area for 2 weeks prior to each tuna sighting), and bottom depth and slope. Mean SST associated with bluefin schools was 18.1°C (±2.8). Schools were located at a mean distance of 19.7 km (±19.6) from SST fronts, and in water masses with an average frontal density of 28.2 m km^?2 (±35.7). Mean bottom depth of detected schools was 139.0 m (±70.3), and mean bottom slope was 0.7% rise (±0.7). A binomial generalized linear model fit to these variables indicated that bluefin are seen closer to fronts than locations in which no tuna were seen. Using simple and partial Mantel tests, we investigated the spatial correlation between bluefin tuna presence and the environmental variables, controlling for spatial autocorrelation. For each day that schools were sighted, we performed 24 Mantel tests, on a combination of response and predictor variables. The spatial relationship between bluefin tuna and SST fronts was inconsistent. Our analysis identified significant spatial structure in the bluefin school locations that had no significant correlation with any of the measured environmental features, suggesting that other untested features, such as prey density, may be important predictors of bluefin distribution in the GOM.     

A comparison of three land-based containment systems for use in culturing green sea urchins, Strongylocentrotus droebachiensis (Müller) (Echinodermata: Echinoidea)

Worldwide, most sea‐urchin populations are in decline. Future market demands will likely be met through aquaculture, which may consist of gonad enhancement of wild‐caught sea urchins. In this context, we examined three land‐based containment systems for suitability in maintaining commercial‐size green sea urchins (Strongylocentrotus droebachiensis). Mortality rate, gonad quantity, gonad quality (colour, brightness, firmness, texture) and cleaning efficiency associated with each containment system were the criteria for comparing the effectiveness of the systems (large raceways, small raceways and washtub tanks). After 6 weeks, urchins maintained in large raceways displayed significantly higher mortality rates than urchins maintained in either the small raceways or washtub tanks (mean±SD: 24.3±10.4%, 8.0±3.5% and 4.3±1.5% respectively). Significant differences in cleaning time were detected among the three systems with washtub tanks requiring significantly more cleaning time (0.11±0.02 min urchin^?1) than the other two systems and large raceways requiring significantly less cleaning time (0.06±0.02 min urchin^?1) than the other two systems. No significant differences in gonad characteristics were detected among the three containment systems. Because of the higher mortality rate in the large raceways and the increased cleaning time required for the washtub tanks, the small raceways were considered to be the best of the three systems tested.     

Changes in the population of American plaice (Hippoglossoides platessoides) off Labrador and northeastern Newfoundland: A collapsing stock with low exploitation

Changes in the population dynamics of American plaice off southern Labrador and eastern Newfoundland (NAFO Subarea 2 and Division 3K) for the period 1977–1994 were investigated. Since the mid 1980s the population has declined dramatically and by the 1990s the stock had essentially collapsed. A higher proportion of what remained of the stock was found in deeper and warmer water than had previously been observed, coincident with a period of unusually cold ocean conditions. During the period of the greatest reductions in population size, the proportion of older, spawning-age fish declined much more rapidly than the younger ages and this decline was accompanied by a slight decrease in mean size at age. Also associated with these observations were declining trends in both size and age at maturity.

Although exploitation has always been low, the fishery for American plaice in NAFO Subarea 2 and Division 3K has been regulated since the early 1970s. For the period investigated the average annual commercial exploitation rate was less than 2% with negligible bycatch in other fisheries. We conclude, therefore, that the collapse of this stock was not a result of fishing mortality. The causal mechanism for the decline remains unclear although unusual environmental conditions during the period continue to be investigated in relation to the observed changes in this population.