绿狐尾藻对不同铵硝配比的生理响应

为了探究高氨养殖废水中一定比例的硝态氮对绿狐尾藻(Myriophyllum aquaticum)生长及生理特征的影响,采用营养液培养,在总氮浓度为15 mmol/L的条件下,以纯铵处理为对照(CK),设置NH+4∶NO-3=3∶1(以N3∶1表示)和NH+4∶NO-3=1∶1(以N1∶1表示)两个处理,研究绿狐尾藻对不同铵硝配比的生理响应。结果显示,培养至第21天,N1∶1处理的相对生物量和相对茎高分别为13.17 g和35.31 cm,显著低于N3∶1(19.59 g,41.78 cm)和CK组(17.82 g, 38.82 cm)(P<0.05)。不同处理下叶片中的氮含量为50.38~58.82 mg/g,显著高于茎中氮含量26.96~35.42 mg/g;但叶片中磷含量5.17~7.38 mg/g显著低于茎中的磷含量8.44~9.88 mg/g(P<0.05)。培养至第21天时,CK、N3∶1、N1∶1处理之间的叶片磷含量(6.76、7.06、7.38 mg/g)差异均达到显著水平(P<0.05),表现为N1∶1> N3∶1>CK。N1∶1和N3∶1处理叶片(7.03、6.75 mg/g)和茎中(6.43、5.04 mg/g)的可溶性糖含量均显著高于CK(3.58、3.27 mg/g)(P<0.05)。N1∶1处理叶片和茎中的叶绿素含量均显著高于CK和N3∶1处理(P<0.05)。研究表明,当处理中添加少量或不添加硝态氮时,有利于绿狐尾藻生长和生物量增加;当处理中硝态氮含量相对较高时,植株叶绿素和可溶性糖含量增加,同时吸收磷的能力增强。绿狐尾藻在不同铵硝配比的水体中均能保持良好的生长状态和较强的氮磷吸收能力,该结果为绿狐尾藻高效净化不同来源的养殖废水提供了理论依据。     

盐酸小檗碱对草鱼补体系统及补体c3作用的研究

为了探究盐酸小檗碱(berberine hydrochloride)对草鱼(Ctenopharyngodon idellus)补体c3的作用机理,用含0、0.06%、0.12%、0.24%盐酸小檗碱饲料投喂草鱼,在第7 d、14 d检测血清中补体c3含量,在第14 d检测肝脏中补体c3 mRNA表达量及急性感染嗜水气单胞菌(Aeromonas hydrophila)后存活情况。并进一步对盐酸小檗碱体内药代动力学和补体消耗试验进行了研究。结果显示;摄食盐酸小檗碱的草鱼血清中补体c3含量和肝脏补体c3 mRNA表达量均显著性增加(P0.05/P0.05),草鱼的存活率与对照组相比也有显著性提高(P0.01)。补体消耗试验结果显示,当盐酸小檗碱在血清中浓度小于5 mg/L时与对照组相比补体消耗不显著,当盐酸小檗碱浓度大于5 mg/L时,补体消耗达到显著性差异(P0.05),表明此时盐酸小檗碱与补体c3直接结合,激活补体。药代动力学实验发现,灌服盐酸小檗碱后,出现双峰现象,两峰值分别是0.243 mg/L和0.117 mg/L,此浓度远远低于与补体c3直接结合量最低量(5 mg/L)。结果表明,盐酸小檗碱对草鱼补体c3的作用机理可能是通过上调补体c3mRNA的表达增加补体c3数量,而非通过直接结合补体c3来改变其活性。     

脊尾白虾14-3-3 基因cDNA 全长的克隆和表达分析

利用本实验室构建的脊尾白虾(Exopalaemon carinicauda)血细胞cDNA文库中脊尾白虾14-3-3巨球蛋白基因EST序列,采用cDNA末端快速扩增(RACE)技术,克隆获得脊尾白虾14-3-3基因c DNA全长,命名为Ec14-3-3。该基因全长2905 bp,包含744 bp的开放阅读框,编码247个氨基酸组成的蛋白质,分子量为27.95 kD,理论等电点为4.65。同源性分析表明,脊尾白虾Ec14-3-3氨基酸序列与斑节对虾(Penaeus monodon)14-3-3的同源性最高,达到98%。荧光定量PCR分析结果表明,Ec14-3-3基因在血细胞、卵巢、肝胰腺等组织中均有表达,其中血细胞中Ec14-3-3相对表达量最高。感染鳗弧菌和WSSV后6 h,脊尾白虾血细胞和肝胰腺中Ec14-3-3基因的表达量较对照组均显著增加(P0.05),且具有明显的时间差异性。本研究结果表明,Ec14-3-3基因在脊尾白虾免疫反应中发挥重要作用。     

Effects of dietary lipid level on growth performance,body composition and digestive enzymes activity of juvenile sea cucumber,Apostichopus japonicus

A feeding trial aimed to determine the effects of dietary lipid level on growth performance, body composition and digestive enzymes activity of juvenile sea cucumber, Apostichopus japonicus. Diets with six crude lipid levels (1.9, 13.8, 29.1, 43.6, 59.6 and 71.6 g kg^?1) were fed to sea cucumbers (initial weights 0.65 ± 0.01 g) at a density of 30 juveniles, once a day. After 60 days, body weight gain (BWG), specific growth rate (SGR), feed intake (FI) and protein efficiency ratio (PER) decreased with increasing dietary lipid levels. The sea cucumbers fed 1.9 g kg^?1 crude lipid showed significantly higher (P < 0.05) BWG than those of the sea cucumbers fed 59.6 and 71.6 g kg^?1 crude lipid. Intestinal protease and lipase activities generally increased with increasing dietary lipid levels. Eicosapentaenoic acid (EPA) content of body walls generally increased with increasing dietary lipid levels. Docosahexaenoic acid (DHA) content of body walls reached the maximum value at a dietary lipid level of 13.8 g kg^?1. N‐3 highly unsaturated fatty acid content followed the same pattern of DHA. According to the growth performance and body composition of sea cucumbers, it can be indicated that the optimum dietary lipid level for juvenile sea cucumbers is between 1.9 and 13.8 g kg^?1.     

Effects of antimicrobial peptides on growth,morphology of foregut villi and related genes mRNA expression in the common carp (Cyprinus carpio)

A 60‐day feeding trial was conducted to examine the effects of different levels (0, 100, 200, 400 and 600 mg/kg) of antimicrobial peptides on growth, protease activity of foregut, the morphology of foregut villi and related genes mRNA expression level in the common carp (Cyprinus carpio). The results showed that the feed of antimicrobial peptides promote common carp growth, and the optimal dosage of antimicrobial peptides is 200–333 mg/kg in the common carp feed. The protease activity of 200 and 400 mg/kg groups were significantly higher than the control and other groups (p < 0.05). The foregut villus height with 100, 200 and 400 mg/kg antimicrobial peptide groups were significantly higher than control group (p < 0.05). The crypt depth of 200 and 400 mg/kg antimicrobial peptide groups were significantly lower than control group (p < 0.05). The ratio of villus height and crypt depth of 100, 200 and 400 mg/kg antimicrobial peptide groups were significantly higher than control group (p < 0.05). The ratio with 600 mg/kg group was significantly lower than the control group (p < 0.05). The IGF‐I gene expression level of 200 mg/kg and 400 mg/kg groups were significantly higher than the control group and 600 mg/kg group (p < 0.05). The IL‐1β gene expression level of 100 mg/kg and 200 mg/kg groups were significantly higher than the control group (p < 0.05). These results indicated up‐regulation of growth and immune related genes in antimicrobial peptides fed common carp. Correlation analysis showed that IGF‐I mRNA and IL‐1β mRNA were positively correlated with SGR. IL‐1β mRNA and FCR were significantly negative correlated. It indicated that growth and immune gene common regulated the growth of the carp under antimicrobial peptides intervention. In conclusion, antimicrobial peptides can improve growth and related genes mRNA expression in the common carp. Further studies using molecular biological technique or immunologic methods are required to conclude that antimicrobial peptides are beneficial in common carp.     

罗氏沼虾育苗水质变动因子及处理技术的研究

针对目前罗氏沼虾育苗生产操作流程,作者研究了育苗池中三态氮、ＣＯＤ 及ｐＨ 等水质变化状况,其特点是这些化学指标无明显的昼夜变化,但ＮＨ３－Ｎ、ＮＯ２－ －Ｎ 含量较高,并呈现波动式上升的趋势,ＣＯＤ 含量较为稳定。从育苗安全性需要出发,还研究了育苗水的预处理模式以及投饵、换水、施药对育苗池水质的影响,结果表明,对河口天然水进行预处理是必要的,作为常规在操作管理上坚持每天彻底排污、适时适量换水、合理用药能有效控制育苗水的水质状况,即使采取高密度（ Ｚ１：２２万尾／ｍ ２）育苗也能获得较高的出苗率     

Phenotypical parameters as a tool to evaluate the immunostimulatory effects of laminarin in Oncorhynchus mykiss

Laminarin is a β‐glucan from the brown algae Laminaria digitata (J.V. Lamour), which can activate innate immune responses. With the aim of developing a strategy to evaluate specific phenotypical parameters of the effects of laminarin in trout innate immunity, we (i) fed fish with laminarin‐supplemented diet (0.2 g kg^?1 day^?1) for 21 days and (ii) treated fish with a single dose of intraperitoneal injected laminarin (0.08 g kg^?1 fish). The evaluation of cellular and humoral immune parameters was established at phenotypic level by the phagocytic activity of headkidney macrophages and detection of inflammatory cytokines in head kidney and gill tissue by indirect ELISA. Results showed that both delivery methods of laminarin produce an increase in the phagocytic activity in head kidney macrophages and a significant increase in the production of TNFα and IL‐8 in gill tissue at day 21. Additionally, some of these parameters were significantly correlated (P < 0.025), which places them as new potential combined markers to detect activation of trout defense mechanisms by laminarin. These results highlight the importance of developing new protocols to quantitate immune parameters, in order to evaluate immunostimulants in fish farming.     

罗非鱼不同组织脂肪酸含量的分析

分别取罗非鱼(Oreochromis niloticus)的肉、脂肪、肝、眼和皮等组织,用氯仿-甲醇(2∶1,v/v)提取,以C17∶0为内标物,利用气相色谱-质谱法(GC/MS)分析了各组织脂肪酸种类及含量。结果显示:罗非鱼各种组织中含有16种脂肪酸,富含多不饱和脂肪酸。脂肪组织中的脂肪酸含量最高,其次是眼睛、肝脏、鱼肉和鱼皮;各组织中饱和脂肪酸(SFA)含量为202~8869 mg/kg,主要为棕榈酸(C16∶0)和硬脂酸(C18∶0);单不饱和脂肪酸(MUFA)含量为293~11219 mg/kg,主要为棕榈油酸(C16∶1)和油酸(C18∶1);多不饱和脂肪酸(PUFA)含量为362~8293 mg/kg,主要是亚油酸(C18∶2)、亚麻酸(C18∶3)和二十二碳六烯酸(DHA,C22∶6);n-6/n-3为1.20~2.46。     

饲料VD3水平对黄颡鱼转录组差异基因表达的影响

为研究饲料中添加维生素D_3对黄颡鱼基因表达的影响,实验基于RNA-Seq技术对用添加不同水平维生素D_3[0(VD0),1243(VD2),22700(VD20)IU/kg]的饲料喂养的黄颡鱼肾脏和小肠的转录组数据进行分析。通过对305 568982条原始reads的筛选和排除,得到了83 265条unigenes,平均长845.38 nt,N50为1620 nt。利用Blast等相关软件对数据进行深度分析,得到功能注释基因共29 224个。根据GO富集分析发现,差异表达基因主要集中在细胞过程、代谢通路、生物调控等通路中,KEGGpathway富集分析结果显示,代谢通路涉及基因最多,有1532个。相对于对照组,添加1243和22700 IU/kg的VD_3(VD0 vs VD2/VD0vs VD20)可得到共同上下调基因共380个,其中上调基因266个,下调基因114个;3种水平下共同上下调的基因共2 1个,其中上调基因4个,下调基因1 7个。研究表明,在饲料中添加不同浓度维生素D_3会对黄颡鱼的生长代谢和生物调控等相关基因表达产生影响,且不同浓度实验组,相关基因的表达存在差异。本实验通过对差异表达基因的后续分析及预测,为黄颡鱼的生长代谢及疾病预防等方面的研究提供了丰富的数据来源。