PCB uptake and accumulation by oysters (Crassostrea virginica) exposed via a contaminated algal diet

Reproductively active oysters were fed daily with 0.2 g algal paste containing 0, 0.1, and 1.0 μg polychlorinated biphenyls (PCBs) (1:1:1 mixture of Aroclor 1242, 1254 and 1260) for either 15 or 30 days, and accumulation of PCBs in different organ tissues and eggs assessed. The effects of PCB exposure on lipid content, lipid class and fatty acid composition were also evaluated. PCBs were accumulated by the oysters and transferred to the eggs. PCB accumulation in oysters was dose, time and tissue dependent. Mean PCB contents were 3150, 1970, and 250 ng/g dry wt., respectively, in the visceral mass, gills+mantle and muscle of oysters fed algal paste containing 1.0 μg PCBs for 30 days. The PCBs in the eggs from the same oysters reached 671 ng PCBs/g dry wt. Feeding oysters with PCB-sorbed algal paste for 30 days significantly increased phospholipid and free fatty acid contents in gills+mantle tissues compared to the same tissues in the undosed control.     

The effect of PCBs on glycogen reserves in the eastern oyster Crassostrea virginica

Recent declines in Chesapeake Bay oyster populations have been attributed to disease, and reduced water quality from pollution. The stress associated with pollutant exposure may reduce energy available for growth and reproduction. Polychlorinated biphenyls (PCBs) are lipophilic contaminants that may potentially affect mobilization of lipid reserves, increasing reliance on glycogen stores, which could otherwise be utilized to supply energy for gametogenesis. Thus, PCBs may indirectly affect glycogen stores in oysters in a deleterious manner. To test for this effect, reproductively inactive oysters were exposed to PCBs by feeding individuals 0.7 g of algal paste containing 0, 0.35, or 3.5 μg PCBs daily for 8 weeks. Additionally, a group of oysters was exposed to PCBs (0, 0.35, and 3.5 μg) plus 0.3 g of non-toxic artificial sediment to examine interactive effects of sediment particles and PCBs. Adductor muscle, mantle, and gonadal tissues were analyzed for glycogen content. Results suggest that glycogen content is reduced in the adductor muscle with increasing PCB exposure, but there are no effects of PCBs in the mantle and gonadal tissues.     

Esterification of vertebrate-like steroids in the eastern oyster (Crassostrea virginica)

The esterification of two model vertebrate steroid hormones – estradiol (E2) and dehidroepiandrosterone (DHEA) – was studied in the oyster Crassostrea virginica. The activity of acyl-CoA:steroid acyltransferase was characterized in microsomal fractions isolated from oyster digestive glands. The apparent K_m and V_max values changed with the fatty acid acyl-CoA used (C20:4, C18:2, C18:1, C16:1, C18:0 or C16:0), and were in the range of 9–17 μM, and 35–74 pmol/min/mg protein for E2, and in the range of 45–120 μM, and 30–182 pmol/min/mg protein for DHEA. Kinetic parameters were also assessed in gonadal tissue. The enzyme saturated at similar concentrations, although conjugation rates were lower than in digestive gland. Preliminary data shows that tributyltin (TBT) in the low μM range (1–50) strongly inhibits E2 and DHEA esterification, the esterification of E2 being more sensitive to inhibition than that of DHEA. Overall, results indicate that apolar conjugation occurs in oysters, in both digestive gland and gonads, at a very similar rate to mammals, suggesting that this is a well conserved conjugation pathway during evolution. Esterification, together with other mechanisms, can modulate endogenous steroid levels in C. virginica, and might be a target for endocrine disrupters, such as TBT.     

单体牡蛎诱导变态的研究

为了得到壳型规则、大小均一的单体牡蛎,作者以葡萄牙牡蛎(Crassostrea angulata)和长牡蛎(Crassostrea gigas)眼点幼虫为材料,从肾上腺素(EPI)浓度梯度、处理时间梯度和眼点幼虫密度梯度3个因素诱导产生不固着变态的单体牡蛎。结果表明,葡萄牙牡蛎用EPI处理24 h的最适浓度为5×10–4 mol/L,不固着变态率为72.8%,在该最适浓度下,最佳处理时间为12 h,不固着变态率为82.7%。长牡蛎用EPI处理6 h的最适浓度为5×10–5 mol/L,不固着变态率为53.2%,在该最适浓度下,最佳处理时间为8h,不固着变态率为56.8%,眼点幼虫密度在80个/m L以下EPI处理效果没有显著性差异。对长牡蛎幼虫进行后续生长测定,结果显示EPI处理组幼虫的壳长、壳高和存活率要高于对照组,表明EPI可能促进牡蛎幼虫变态长出次生壳并提高其生存能力。     

Response of a benthic suspension feeder (Crassostrea virginica Gmelin) to three centuries of anthropogenic eutrophication in Chesapeake Bay

Biogenic reefs built by oysters and other suspension feeders are vital components of estuarine ecosystems. By consuming phytoplankton, suspension feeders act to suppress accumulation of organic matter in the water column. Nutrient loading increases the rate of primary production, thereby causing eutrophication. As suspension feeders consume more organic matter from increasing abundance of phytoplankton, their rate of growth should also increase if they are food limited. We show here that the eastern oyster, Crassostrea virginica (Gmelin), from St. Mary's and Patuxent rivers, Chesapeake Bay, grew faster during anthropogenic eutrophication relative to C. virginica before eutrophication. Growth of shell height, shell thickness and adductor muscle increased after eutrophication began in the late 18th century. After 1860, growth decreased, perhaps reflecting the negative effects of hypoxia, harmful algal blooms, disease and fishing on oyster growth. These results are consistent with the view that an increasing supply of phytoplankton resulting from eutrophication enhanced growth of C. virginica between 1760 and 1860, before oyster reefs were degraded by destructive fishing practices between 1870 and 1930. Alternative factors, such as changes in water temperature, salinity, and fishing are less likely to be responsible for this pattern. These results have implications for restoration of oyster reefs in order to mitigate the effects of eutrophication in estuaries, as well as the paleoecological relationship between suspension feeders and paleoproductivity.     

不同长牡蛎群体对海水酸化的生理响应差异分析

海水酸化暴露可对海洋生物产生多层面的影响。本研究以潮间带野生与潮下带养殖长牡蛎(不同生境背景)的不同组织(鳃、外套膜及消化腺)为研究对象,分析在室内调控p CO2模拟海水酸化暴露条件下,其基础代谢活动、能量代谢以及氧化应激相关指标的变化情况。结果显示:海水酸化暴露后,两种长牡蛎(Crassostrea gigas)的基础代谢过程均受到了一定抑制作用且受影响程度差异明显。潮间带野生与潮下带养殖长牡蛎的关键生理过程(能量代谢及氧化应激)对海水酸化暴露存在不同的响应变化,表明两种长牡蛎应对海水酸化的调节机制可能存在差异。依据PLS-DA分析结果显示,在所有生理指标中,对样本的差异贡献较高(VIP值>1)的指标为:SDH、AST、ATPase、ATP含量、糖原含量、CAT、GST及SOD,表明海水酸化暴露后,在两种长牡蛎的3种组织中上述指标的响应变化程度更大。综合评价分析多个生理指标的整体变化揭示:在海水酸化暴露条件下,潮间带野生长牡蛎比潮下带养殖长牡蛎对海水酸化的生理响应更为剧烈;相比于鳃及消化腺组织,长牡蛎外套膜组织可能受影响更大。     

糖原含量与糖原磷酸化酶基因(GPH)和己糖激酶基因(HK)的表达在福建牡蛎生殖周期中的相关性分析

Glycogen, a polymer of glucose, is an important means of storing energy. It is degraded by glycogen phosphorylase (GPH) and hexokinase (HK), glycogen phosphorylase, and hexokinase cDNAs (Ca-GPH andCa-H...     

香港巨牡蛎ISSR-PCR反应体系的建立及优化

以香港巨牡蛎（Crassostreahongkongensis）基因组DNA为模板,采用正交设计及单因素比较试验,分别对Mg^2＋、dNTPs、引物、TaqDNA聚合酶和DNA模板浓度的PCR原料进行优化,并通过温度梯度PCR,筛选适宜的退火温度。确立了香港巨牡蛎的最适ISSR—PCR反应体系：25μL反应体系中含PC...     

温度与盐度对福建牡蛎与长牡蛎、熊本牡蛎、香港巨牡蛎种间杂交子代孵化及其早期生长成活的影响

本研究以野生福建牡蛎(A)为亲本一方, 分别与野生长牡蛎(G)、熊本牡蛎(S)、香港巨牡蛎(H)进行杂交, 除A♀×S♂、A♀×H♂两组杂交不育外, 共得到AG(A♀×G♂)、GA(G♀×A♂)、SA(S♀×A♂)、HA(H♀×A♂)、AA(A♀×A♂)5个不同组合, 并研究不同盐度与温度对其幼虫孵化率、早期生长及成活率的影响。结果表明, 盐度25时AG、GA、SA、HA、AA五组孵化率分别达到最大值78.33%、75.00%、63.33%、55.00%、71.67%; 温度低于25℃时, GA孵化率高于其他组, 温度高于25℃时AG孵化率最大, 其中25℃时达到最大孵化率83.33%; 不同盐度条件下D型幼虫经过7 d培育, HA在盐度20时壳高达到最大值97.8 µm, 成活率46.67%, 在盐度为35时第5天全部死亡。AG、GA、SA、AA四组在盐度为25时壳高有最大值, 分别为124.7 µm、121.2 µm、93.7 µm, 111.8 µm; 不同温度条件下D型幼虫经过7 d培育, 在温度为25℃时各组壳高和成活率均有最大值, 大小排列依次为AG>GA>AA>HA>SA, 各组成活率随温度升高呈现出先增大后减小的趋势, AG在25℃时成活率在所有组中最高, 为88.89%; 孵化后在适宜温度盐度条件下连续培养16 d, 不同组之间壳高表现为: AG > GA> AA > HA > SA, AG壳高、壳高SGR及成活率始终大于其他组, 而SA、HA两组壳高、壳高SGR及成活率始终小于自交组AA。研究表明, 相较于福建牡蛎与熊本牡蛎、香港巨牡蛎的杂交, 以及福建牡蛎的自交, 福建牡蛎与长牡蛎的杂交子代表现出明显的生长及存活优势, 生长潜力较好, 且AG优于GA; AG组在盐度25、水温25℃条件下生长速率最大, 成活率最高。研究结果可为福建牡蛎育苗行业良种选育提供参考依据。     

近江牡蛎鳃细胞的原代培养

采用改良的DMEM(HG)培养基,建立了近江牡蛎(Crassostrea hongkongensis)鳃细胞体外培养技术,包括鳃组织块培养法和胰酶消化培养法。结果表明,组织块培养法接种6 h后,细胞开始从组织块中迁出,细胞形态较小,呈圆形、椭圆形或多边形,直径3～6μm。培养至3 d时,细胞在组织块周围形成生长晕。培养至6 d时可进行细胞传代,本次实验细胞已传至第6代。胰酶消化法接种约2 h后,细胞逐渐贴壁,从形态上主要分为两类,一类为小型细胞,形态为圆形、椭圆形或多边形,直径3～6μm,数量多,增殖速度较快;另一类为大型细胞,形态为圆形、椭圆形或多边形,直径10～20μm,部分细胞内部含有颗粒,数量较少,增殖速度较慢。培养至2 d时可进行细胞传代,传代培养物中的优势细胞皆为小型细胞。本次实验细胞已传至第6代。     

海滨锦葵种子含油量的遗传分析

海滨锦葵(Kosteletzkya virginiva)是重要的能源油料植物,对海滨锦葵种子含油量进行遗传分析具有重要意义.以高油海滨锦葵品种KV23(P1和低油海滨锦葵品种KV16(P2)及其杂交组合的F1、F2群体为材料,以单株种子含油量为指标,应用P1、P2、F1和F2 4个世代的数量性状主基因+多基因混合遗传模型联合分离分析方法,对海滨锦葵种子含油量进行了遗传分析.结果表明,海滨锦葵种子含油量可能由两时加性-显性-上位性主基因+加性-显性多基因共同控制,主基因遗传率为49.87%,主基因和多基因共同解释的遗传率为80.14%.     

长牡蛎外套膜色遗传规律初步研究

比较了长牡蛎(Crassostrea gigas)不同外套膜色家系的受精率与孵化率,不同生长阶段的生长与存活及外套膜色在子代个体中的分布规律。结果表明,不同家系的卵径、受精率和孵化率无显著差异(P0.05);壳高在60和150日龄稚贝阶段略有差异(P0.05),在350和450日龄成体阶段差异不显著(P0.05);不同生长阶段的存活率差异不显著(P0.05)。外套膜纯黑BB家系的子代外套膜基本全部为4级着色;外套膜纯黄YY1家系子代的外套膜基本全部为1级着色,YY2家系子代的外套膜黄色与黑色比例约为3∶1,符合孟德尔分离定律。由此推测长牡蛎外套膜色可能由一个主效基因控制,且黄色对黑色具有显性上位效应。研究结果可以为长牡蛎外套膜色的人工选育提供理论依据。     

响应面法优化香港巨牡蛎蛋白酶解工艺的研究

以水解度为指标,比较了6种蛋白酶对香港巨牡蛎(Crassostrea hongkongensis)的水解效果。在单因素考察实验的基础上,采用Box-Behnken模型响应面设计,建立香港巨牡蛎蛋白酶解的二次回归模型,确定最佳的水解条件。实验结果表明,海产品专用蛋白酶对香港巨牡蛎蛋白的水解效果优于其他蛋白酶,该酶的最佳水解条件为:酶与底物比为5.93%,酶解时间为4.07 h,酶解温度为54.6℃。在此条件下,海产品专用蛋白酶酶解香港巨牡蛎蛋白的水解度达到25.73%±2.39%。本研究为香港巨牡蛎的开发和利用提供了理论依据。     

香港巨牡蛎对3种微藻的选择性摄食研究

从微藻粒径大小、饵料密度以及营养价值角度,选取牟氏角毛藻（Chaetoceros muelleri）、亚心形扁藻（Platymonas subcordiformis）、球等鞭金藻（Isochrysis galbana）分别作为硅藻、绿藻和金藻的代表,开展了不同规格的香港巨牡蛎对3种微藻在同等密度、同等生物量混合条件下的摄食选择性研究。结果显示,香港巨牡蛎在两种不同混合藻条件下对3种不同微藻摄食选择性有显著性差异（P<0.01）。3种藻相同密度混合条件下,大、中、小3种规格香港巨牡蛎更倾向摄食粒径较大的亚心形扁藻,且贝类规格越大选择倾向性越低,摄食选择效率分别为0.32、0.35、0.48,对球等鞭金藻无明显选择性。3种藻等生物量混合条件下,香港巨牡蛎对亚心形扁藻和球等鞭金藻的摄食有选择性,且规格大小对选择效率影响显著（P<0.01）,个体越小选择性越强,小规格香港巨牡蛎倾向滤食亚心形扁藻（P<0.01）,大、中规格更倾向摄食粒径较小的球等鞭金藻。两种混合条件下,香港巨牡蛎都对牟氏角毛藻都有负的选择效率,而且牟氏角毛藻密度越高负值绝对值越大。微藻的细胞大小、形状、营养价值以及香港巨牡蛎的个体大小都对摄食选择性有着明显的影响,香港巨牡蛎的摄食选择性在现场海区中还明显受到生境及食物条件的限制。     

香港牡蛎二倍体及三倍体幼虫生物学特征比较

为了评估香港牡蛎二倍体及三倍体幼虫生物学特征,于2013～2015年,在湛江和北海两地比较了二倍体及三倍体幼虫生长、存活及变态差异,计算了三倍体优势率,评估了倍性与环境效应互作对其表型性状的影响。结果表明:三倍体D形幼虫显著大于二倍体幼虫,之后,伴随着个体增长,生长优势逐渐增大,15日龄时的三倍体优势率为8.9%～9.1%。三倍体幼虫存活力与二倍体相当,未出现大量致死现象。三倍体变态规格显著大于二倍体,其优势率为5.1%～5.3%;三倍体变态时间显著低于二倍体,但其变态率与二倍体相当,不存在显著差异(p>0.05)。经过双因子分析模型的方差分析检测,发现倍性和环境效应都会对香港牡蛎幼虫生长产生显著影响(p<0.001),环境是影响幼虫存活率及变态的主要因子(p<0.05)。     

长牡蛎两种engrailed同源基因的鉴定及其与贝壳发育相关性的研究

engrailed基因属于同源异形基因家族成员,在许多动物的分节、附肢发育、神经系统发育和贝壳形成过程中发挥作用。本研究克隆了长牡蛎两个engrailed同源基因,命名为cgi-eng1和cgi-eng2。序列分析表明,两个基因均具备典型engrailed基因保守的5个EH结构域。利用整装原位杂交技术检测了cgi-eng1和cgi-eng2在贝壳形成的关键时期早期D形幼虫时期的表达情况。结果显示,cgi-eng1和cgi-eng2 m RNA高表达于贝壳外缘,可能与早期贝壳形成过程有关。此外,两个基因的在贝壳外缘的表达模式亦有区别,提示两个基因的功能可能存在一定程度的分化。本研究首次系统鉴定了长牡蛎engrailed基因的成员,并发现它们可能均参与幼虫贝壳形成,研究结果有助于加深对贝类早期发育及贝壳形成的理解。     

广西钦州湾近江牡蛎体有机氯农药和多氯联苯的残留水平与分布

对广西钦州湾近江牡蛎(Crassostrea rivularis)体有机氯农药(HCHs和DDTs)和多氯联苯(PCBs)的残留水平进行监测与分析。结果表明:近江牡蛎体中HCHs、DDTs和PCBs的质量比分别为ND～0.23μg/kg、0.01～0.21μg/kg和ND～2.3μg/kg;检出率分别为45%、100%和95%,3种污染物含量符合国家《海洋生物质量》第一类标准和无公害水产品质量要求。相对国内其他海区而言,钦州湾近江牡蛎受HCHs、DDTs和PCBs污染程度处于低污染水平。     

Depuration dynamics of oysters (Crassostrea gigas) artificially contaminated by Salmonella enterica serovar Typhimurium

The State of Santa Catarina produces the greatest quantity of edible mollusks in Brazil. To guarantee sanitary qualify, mollusk cultures should be monitored for contamination by pathogenic microorganisms. A self-purification or "depuration" system that eliminates Salmonella enterica serovar Typhimurium contamination from oysters has been developed and evaluated. The depuration process occurred within a closed system, in which 1000 L of water was recirculated for 24 h. The water was sterilized with ultraviolet (UV) light, chlorine, or both together. Oysters (Crassostrea gigas) artificially contaminated with S. typhimurium were harvested every 6 h. Samples of oyster tissue were excised and both the presence and numbers of bacteria were determined. Combined UV light and chlorine treatments resulted in total elimination of bacteria within 12 h. Polymerase chain reaction detected bacteria in water exposed to the three treatments. This pioneering study is the first of its kind in Brazil and represents a major contribution to commercial mollusk culture in this country.     

长牡蛎dpp同源基因的克隆及其在贝壳发生中的功能研究

dpp是转化生长因子β家族的重要成员,在胚胎发育中作为形态发生子参与体轴形成及附肢发育等过程。为了研究牡蛎dpp同源基因在贝壳发生中的作用,揭示贝壳发生的分子机理,作者克隆了一种长牡蛎(Crassostrea gigas)的dpp基因,命名为cgdpp。序列和进化分析显示,cgdpp分子中不同位点的氨基酸残基变异程度不同。作者用整装原位杂交的手段研究了dpp同源基因在长牡蛎早期发育中的表达情况。结果表明,dpp同源基因参与了从贝壳形成区开始分化到早期贝壳形成的全过程。在担轮幼虫(Trochophore)中,dpp同源基因似乎调控了贝壳的形状与扩张速度;在早期D形幼虫中,dpp同源基因表达量突然下降至痕量水平,并与贝壳发育区无明显的关联,提示dpp同源基因可能仅仅参与了贝壳的发生,不参与其进一步的发育过程。     

多肽菌素在长牡蛎人工育苗生产中的应用

多肽菌素作为一类具有抗菌活性的生物短肽,是一种可以替代抗生素的生物肽。为了推广多肽菌素在水产养殖中的应用,本研究以长牡蛎(Crassostreagigas)人工育苗为基础,以添加2×10_–6 mol/L青霉素钠为对照组,以添加5个不同浓度(2×10_–6、4×10_–6、6×10_–6、8×10_–6、10×10_–6 mol/L)多肽菌素为实验组,研究多肽菌素在长牡蛎人工育苗生产中的应用。结果显示:(1)在长牡蛎种贝培育方面,培育前8天各组差异不显著(P> 0.05),培育后期添加6×10_–6 mol/L多肽菌素组存活率均最高,对照组最低,且培育至32天时该实验组均显著高于其他组(P<0.05)。(2)在长牡蛎幼虫培育方面,幼虫壳高日增长量在添加6×10_–6 mol/L多肽菌素组中最大,为11.57μm/d,而对照组仅为9.61μm/d;幼虫存活率方面对照组始终最低,而添加6×10_–6 ...