Cu^2+胁迫对魁蚶生理生化和组织结构的影响

采用生物毒性测试的方法,研究了持续暴露96 h不同浓度Cu2+胁迫对魁蚶生理代谢、组织结构及酶活性的影响。实验设置0.01、0.05、0.10、0.50和1.00 mg/L组5个浓度梯度,不加Cu2+的正常海水作为对照。结果显示,Cu2+胁迫对魁蚶耗氧率(OR)、排氨率(NR)、氧氮比(O∶N)均有显著影响,并且与暴露时间和浓度有关。初次暴露时,OR,NR和O∶N都迅速下降,最低值都出现在1.00 mg/L浓度组,分别为(0.005±0.001)mg/(g·h)、(0.5±0.05)μmol/(g·h)、0.7±0.1,仅为对照组的1%、29%和3%。暴露72 h内Cu2+胁迫均使魁蚶代谢率不同程度下降,之后不同处理组出现差异分化。暴露96 h时,魁蚶对低浓度Cu2+暴露表现出适应,0.01 mg/L处理组呼吸代谢恢复至正常水平,O∶N与对照组无差异,组织结构也未见明显损伤;浓度超过0.05 mg/L处理组魁蚶生理代谢及组织结构受显著影响,O∶N大多降至9以下,出现鳃丝受损和组织结构散乱等明显损伤;0.10 mg/L处理组魁蚶体内ACP和ALP活性在鳃组织中增强,肝脏中受到抑制,而GPX和GST活性增强。研究表明持续暴露96 h Cu2+浓度≥0.05 mg/L环境显著影响魁蚶生理代谢及组织结构,0.10 mg/L Cu2+浓度显著影响魁蚶组织中ACP、ALP、GPX和GST的活性。研究结果为认知魁蚶等滩涂贝类对Cu2+胁迫的响应机制提供基础数据,为预防滩涂潜在重金属污染风险及生物修复提供参考。

魁蚶C型凝集素基因cDNA的克隆及表达分析

通过cDNA末端快速扩增(Rapid amplification of c DNA ends,RACE)技术克隆得到魁蚶(Scapharca broughtonii)C型凝集素(C-type lectin,Sb-Lec1)基因,该基因全长为700 bp,其中,5′-UTR为29 bp,3′-UTR为167 bp,开放阅读框长度为504 bp,编码167个氨基酸,包括长度为23个氨基酸的信号肽序列、129个氨基酸的糖识别结构域(CRD)以及参与二硫键形成的6个半胱氨酸。预测蛋白分子量为19.11 k Da,理论等电点为4.74。多序列比对结果显示,Sb-Lec1基因CRD编码的氨基酸序列与长牡蛎(Crassostrea gigas)、紫贻贝(Mytilus galloprovincialis)和海湾扇贝(Argopecten irradians)C型凝集素的同源性分别为38%~40%、34%~35%和38%~39%,Sb-Lec1基因编码的氨基酸序列与其他物种的凝集素基因具有相似的结构,均含有形成二硫键的4个保守半胱氨酸。系统进化分析结果显示,魁蚶先与贝类聚为一支,再与脊椎动物聚在一起,表明魁蚶Sb-Lec1基因在进化树上的位置与其传统分类所处位置一致。采用荧光定量PCR技术,检测了Sb-Lec1基因在组织中的表达情况,发现其在肝胰腺、血淋巴、鳃、外套膜、闭壳肌、斧足中均有表达,其中,肝胰腺表达量最高。同时,分析了Sb-Lec1基因在鳗弧菌(Vibrio anguillarum)刺激下的mRNA表达量变化情况。结果显示,与对照组相比,菌刺激组Sb-Lec1基因mRNA在各检测组织中的表达量均显著上调(P<0.05),随着刺激时间的延长,表达量呈先升高后降低的趋势。本研究表明,魁蚶Sb-Lec1基因在机体免疫防御方面发挥重要功能。

短时间高盐处理对脆江蓠光合生理生化指标的影响

脆江蓠(Gracilaria chouae)藻体脆性大,短时间高盐海水浸泡可使其软化,有利于规模夹苗生产。采用实验生态学方法研究短期高盐胁迫对脆江蓠光合生理及生化组成的影响,以探讨高盐对其光合系统的损伤及其恢复效果。实验设置 5 个盐度梯度(40、45、50、55、60),自然海水作为对照(盐度 33),分析盐度处理 1 h 内脆江蓠的失水率及藻体软化程度;同时研究了高盐处理 0.5 h 及不同恢复时间(12 h、24 h)对脆江蓠 pH 补偿点、光合作用荧光特性、光合作用产氧量(RO)和光合色素组成等光合生理生化指标的影响。结果表明,脆江蓠在盐度 50~55 的海水中浸泡 0.5 h 效果较好,此时藻体软化,并且经过 24 h 恢复后,藻体光合作用参数可恢复到对照组水平,该处理条件可以用于脆江蓠生产。高盐(盐度 40~60)处理脆江蓠 0.5 h,随盐度增加,脆江蓠 RO 值呈波动下降(P<0.01)、pH补偿点和光合效率 Y(II)值逐渐降低(P<0.05)。PE 含量随盐度增加而增加(P<0.01);恢复 24 h 后最大光合效率 Fv/Fm和 Y(II)值基本恢复正常水平(P>0.05), Chl a、Car、PE 和 PC 含量均恢复到与对照组无显著性差异水平(P>0.05)。本研究旨在为提高脆江蓠规模化养殖夹苗效率提供理论依据。

Assessing the effects of oyster/kelp weight ratio on water column properties: an experimental IMTA study at Sanggou Bay, China

Integrated Multi-Trophic Aquaculture(IMTA)is an effective method for sustainable aquaculture as species from different trophic levels could reduce negative effects from fed species in the environment.A proper proportion of different trophic species in an IMTA system could improve the aquaculture production and environmental sustainability.At present,research on the proper proportions for farming species is scarce.We investigated the effects of IMTA modes of oyster(Crassostrea gigas)and kelp(Saccharina japonica)in different weight ratios on water quality and carbonate system in a closed enclosure experiment for three days in the Sanggou Bay in Shandong Province,China,in December 2017.Nine collocation modes in oyster:kelp weight ratio were tested showing as 24:3,24:2,24:1,16:3,16:2,16:1,8:3,8:2,and 8:1.The water parameters were determined at 17:00 on Day 1(D1),and 6:00 and 17:00 on Days 2(D2)and 3(D3).As two-way ANOVA showed,all increased parameters(dissolved oxygen(DO),pH,chl a,the carbonate system and pCO2)were significantly related to oyster-kelp modes,and interaction between modes and time were also significant(P<0.05).On the 3th day,the 8:3 mode was the highest in DO,pH,chl a,CO32-(P<0.05),and dissolved inorganic carbon(DIC),HCO3-,CO2,and pCO2 were the lowest(P<0.05).According to previous references and the results of this study,the appropriate oyster:kelp proportion at the beginning of winter is from 8:2 to 8:3.The results of this study may help government to optimize the aquaculture structure of Sanggou Bay.

Response of Yesso scallop Patinopecten yessoensis to acute temperature challenge: physiological and biochemical parameters

Water temperature is generally considered to be a major factor af fecting the physiological and biochemical activities of marine bivalves. Here, the physiological and biochemical responses of Yesso scallop, Patinopecten yessoensis, to acute water temperature changes in summer were studied. Scallops were transferred directly to a lower temperature( T dec treatment)(from 23°C to 15°C) or to a higher temperature( Ti nc treatment)(from 15°C to 23°C) for 72 h, respectively. Results showed that the oxygen consumption and ammonia-N excretion rates of P. yessoensis decreased signi?cantly in the T dec treatment but increased dramatically at 6 h in the Ti nc treatment( P <0.05). In the T dec treatment, hepatopancreas antioxidant enzyme activities, superoxide dismutase(SOD) and catalase(CAT) activities, increased substantially within 72 h( P <0.05). However, a signi?cant decrease in CAT activity was found at 12 h in the Ti nc treatment( P <0.01).A signi?cant enhancement of acid phosphatase(ACP) activity and malondialdehyde(MDA) content was detected when scallops were acutely exposed to a temperature of 15°C. The levels of Cu/Zn-SOD gene expression in their gills up-regulated signi?cantly in response to acute temperature changes( P <0.01). These data suggest that acute temperature change af fects physiological and biochemical functions, and improve our knowledge of P. yessoensis under conditions of thermal stress.