草鱼肠系膜脂肪组织高质量总RNA提取方法的研究

基于传统的酸性酚—异硫氰酸胍—氯仿一步提取法,比较分析多种优化操作步骤,摸索出一种高质量提取体质量为80~150 g草鱼肠系膜脂肪组织总RNA的改良方法。试验结果显示,相较于肝脏、脾脏、肠道等脏器组织,草鱼肠系膜脂肪组织RNA丰度低,且极易在样品前处理阶段出现顽固性降解问题。探索发现,将取样量增至约30 mg,可提升RNA产量以满足常规试验需求。针对降解难题,改良常规的样品前处理技术流程,采用鲜样液氮速冻,冻样直接放入TRIzol试剂中裂解,并即刻进行长时间机械匀浆,时长约3 min等核心操作步骤,可显著降低脂肪组织样品RNA的降解。Agilent生物分析仪检测结果显示,改良方法提取的草鱼肠系膜脂肪组织RNA完整度高,关键RIN值为8.7~9.0。研究推测,长时间机械匀浆所形成的持续剪切冲击力或许有助于TRIzol试剂中的异硫氰酸胍等成分突破油滴阻碍而有效抑制内源性RNA酶。本方法提取的草鱼脂肪组织总RNA质量可满足高通量转录组测序要求。     

多营养层次生态养殖模式简析

系统介绍了多营养层次生态养殖模式在淡水养殖及海水养殖中的应用与发展,对比了国内外有关该养殖模式发展方向的异同,简述了该养殖模式通过提高物质和空间利用率、改善养殖环境的原理以及在我国推广应用情况,并汇总了我国沿海省份近年来开展该养殖模式选用的品种搭配及产量产值。指出了多营养层次生态养殖模式还存在的问题,对今后发展提出了建议。     

Nuclear transfer in loach (Paramisgurnus dabryanus Sauvage) by cell-to-cell electrofusion

To study nuclear transfer in the loach (Paramisgurnus dabryanus Sauvage), blastula and gastrula cells were fused with UV-inactivated oocytes by cell-to-cell electrofusion. To facilitate nuclear transfer, blastula and gastrula cells were cultured or incubated at 4 °C in different solutions. TC-199 medium supplemented with 20% calf serum was the best culture solution, and effectively retained the totipotence of blastula or gastrula cells for up to 10 days. It was found that gastrula cells incubated at 4 °C had the same totipotence as blastula cells. The optimal UV dosage for inactivation of the oocyte chromatin was 180–240 mJ cm⁻². Electrofusion was carried out in a cone-shaped fusion chamber, which permitted the recipient oocyte and the donor blastula cell to contact one another. The electrofusion procedure resulted in a 10% success rate of normal-appearing fish. Genetic analysis indicated that the nuclear material originated from the donor cell (blastomere) and the oocyte pronucleus did not take part in development.     

Disaccharide combinations and the expression of enolase3 and plasma membrane Ca2+ ATPase isoform in sturgeon sperm cryopreservation

Acipenser sinensis and Acipenser dabryanus are critically endangered species, so germplasm conservation via cryopreservation of sperm is necessary. Disaccharides can act as membrane‐impermeable cryoprotectants, and enolase3 (ENO3) and plasma membrane Ca²⁺ ATPase isoform (PMCA2) are proteins associated with sperm quality. We considered seven characteristics of sperm quality in cultured brood stock from A. sinensis and A. dabryanus. We tested use of sucrose or trehalose alone and in combination at different concentrations for cryopreservation of A. dabryanus sperm. A low concentration of sucrose plus trehalose (S₁₅T₁₅) was optimal. Mixing of the extender with sucrose, lactose, or trehalose alone or with pairwise mixtures revealed that a mixture of lactose and trehalose (L₁₅T₁₅) gave the best results for both A. sinensis and A. dabryanus. Enolase3 and PMCA2 expression levels were measured in cryopreserved A. sinensis sperm via Western blotting. Relative ENO3 and PMCA2 expression levels were examined, and the relationship between disaccharide composition, sperm quality and protein expression was explored in A. sinensis. The results showed that relative ENO3 and PMCA2 expression levels were the highest at L₁₅T₁₅ in cryopreserved A. sinensis sperm. There were significant positive correlations between ENO3 expression and percentage membrane integrity, and between PMCA2 expression and sperm motility parameters (percentage of motile sperm, curvilinear velocity, straight‐line velocity and average path velocity; p < .05) in cryopreserved A. sinensis sperm. Our results indicate the optimal disaccharide combination and concentrations for cryopreservation of A. sinensis and A. dabryanus sperm and suggest that ENO3 and PMCA2 expression levels could serve as a valuable indicator of sperm quality in A. sinensis.     

水产养殖尾水初沉区中净化材料的应用对微生物酶活性的影响

通过检测除氮型改性凹凸棒土（Al@TCAP-N）和火山石的内部及周围水体有关氮、磷、有机物分解的酶活性和水质指标，利用高通量测序对材料和水体中的细菌和真菌进行分析，以研究尾水处理系统初沉区中水质净化材料处理尾水营养盐的机制。结果表明，Al@TCAP-N和火山石能增加其周围水体微生物碱性磷酸酶（AKP）、硝酸盐还原酶（Nar）活性；火山石能在早期（0~6 h）提高其内部有机磷水解酶（OPH）、氨单加氧酶（AMO）的活性；Al@TCAP-N能在后期（36~48 h）增加其内部脱氢酶（DHO）的活性；净化材料相互对比发现，火山石内部的酶活性整体高于Al@TCAP-N内部的酶活性；本试验表明水质处理最佳时间为36 h，总氮（TN）、总磷（TP）、高锰酸盐指数（COD_Mn）的去除率分别为22.61%、9.52%和22.16%。在实际应用过程中，可通过Al@TCAP-N的吸附和火山石负载的微生物的双重作用降低水中营养盐的含量。浮霉菌门（Planctomycetes）、变形菌门（Protepbacteria）、拟杆菌门（Bacteroidetes）和壶菌门（Chytridiomycota）分别是材料表面微生物细菌和真菌中的主要门类。火山石能在净化初期促进有机磷化合物分解和硝态氮（NO₃^--N）转化为亚硝态氮（NO₂^--N），Al@TCAP-N在净化后期促进有机物的分解。Al@TCAP-N和火山石能促进水体中含磷化合物的分解和氮的转化。     

循环水养殖系统水净化装备技术研究

循环水养殖系统为当前水产养殖先进生产力的发展方向,其具养殖水环境高度可控、产品质量安全可靠、节水、节地等特点,符合国家转变经济增长方式,节能减排的战略需求。循环水养殖水净化装备是循环水养殖系统核心组成。其为循环水养殖生产中的水产养殖对象提供良好水生态环境,提高水产养殖品产量及质量,提升资源利用率。本文以循环水养殖系统水净化典型工艺为主线,对其主要环节包括悬浮物去除、溶解性有机物降解、杀菌消毒、增氧、调温等所涉及的装备技术进行探讨及总结,并提出发展建议,以期为精准、高效、节能循环水养殖系统构建,推进其进一步应用提供支持。     

聚乙烯单丝产品质量现状与分析

本文主要介绍了当前我国聚乙烯单丝产品质量现状、产品质量不合格原因，并对产品质量现状提出一点看法。     

南美白对虾池塘养殖水质智能控制系统设计与开发

南美白对虾池塘养殖对于水体质量有着较为严苛的要求,残饵及生物排泄物的增多会大量消耗水体溶解氧,进而打破水体动态平衡,导致NH₃-N、NO₂-N等毒害成分升高,引起南美白对虾病害甚至导致死亡,造成养殖户经济损失并且阻碍了南美白对虾养殖产业的健康发展。针对上述问题,笔者研究并开发一种能够实时监测养殖池塘水质,并能根据南美白对虾生长习性智能控制增氧机、水泵等养殖设备的南美白对虾池塘养殖水质智能控制系统。系统具有控制智能、运行可靠、集成度高等特点,在养殖生产试验中取得了良好效果,保证了南美白对虾养殖过程的安全,有效降低了南美白对虾池塘养殖风险。     

氨氮胁迫对四鼻须鲤鱼肝脏GST酶活力的影响

实验采用生态毒理学方法,以NH4Cl为实验药物,设置0、50、150和250mg/L实验浓度。通过对野生和养殖群体的四鼻须鲤鱼氨氮对肝脏谷胱甘肽巯基转移酶（GST）活力的影响,比较两个群体抗氨氮能力。结果表明：在相同的氨氮浓度和胁迫时间下,四鼻须鲤鱼养殖群体GST酶活力的变化趋势与野生群体大致相同。除250mg/L浓度组在胁迫5d和10d时,野生群体酶活性略低于养殖群体外,在其余时间点的野生群体酶活性均高于养殖群体,而且,在高浓度氨氮胁迫下,野生群体肝脏的GST酶活性达到峰值的时间比养殖群体更短,显示出较强的抗氨氮能力。