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。     

草鱼MYH mRNA表达量分析中采用的内参基因稳定性比较

本研究分别以β-actin、18SrRNA和GAPDH为内参基因,采用实时荧光定量PCR对草鱼早期发育时期肌球蛋白重链（myosin heavy light,MYH）基因的mRNA表达量进行分析,并比较不同内参基因对MYH基因mRNA表达水平检测结果的准确性。研究结果表明,以β-actin和GAPDH作为内参,MYH基因mRNA表达水平完全一致,其表达量从原肠到仔鱼阶段逐次递增,仔鱼与原肠期阶段相比表达量差异显著;当采用18S rRNA作为内参时,MYH基因mRNA在发育阶段的表达量呈不稳定状态。因此,β-actin和GAPDH均可作为内参基因,用于草鱼早期发育中MYH基因mRNA的相对定量研究;而18S rRNA作为内参时,可能会对检测结果造成偏差。本研究不仅准确的揭示了草鱼MYH基因mRNA的表达特征,并且为荧光定量PCR技术在鱼类基因表达研究方面提供了有价值的参考。     

Seminal plasma composition and its relationship with physical spermatological parameters of Grass carp (Ctenopharyngodon idella) semen: with emphasis on sperm motility

In this research, the mineral and organic composition of the seminal plasma, physical spermatological parameters and their physiological relationships were investigated in grass carp (Ctenopharyngodon idella). The seminal plasma contained 98.14±5.23 mM L^?1 (Na⁺), 380.85±25.95 mM L^?1 (K⁺), 30.25±4.96 mg dL^?1 (Ca²⁺), 19.16±1.70 mEq L^?1 (Mg²⁺), 1.36±0.11 mg dL^?1 glucose, 0.37±0.08 g dL^?1 total protein, 12.02±1.18 mg dL^?1 cholesterol, 14.85±1.50 mg dL^?1 triglyceride and 43.5±9.56 mg dL^?1 urea. The following spermatological parameters were found: sperm volume 14.44±1.16 mL, sperm motility 80.60±1.55%, movement duration 67.68±4.32 s, density 15.43±0.72 × 10⁹ mL^?1, total density 337.43+45.86 × 10⁹ and pH 7.24±0.17. The Na⁺ and Ca²⁺ ions correlated negatively with spermatozoa motility (r=?0.453, P>0.05 and r=?0.192, P>0.05) respectively. The K⁺ ion correlated positively with spermatozoa motility (r=0.545, P>0.05). But a statistically significant correlation was not observed between sperm motility and seminal plasma parameters. The following correlations were observed between mineral and organic components. The Mg²⁺ was positively correlated with glucose and cholesterol (r=0.692, P<0.05 and r=0.680, P<0.05) respectively. A highly significant positive relationship was also found between Mg²⁺ and total protein (r=0.837, P<0.01). On the other hand, a significantly negative relationship was found between Ca²⁺ and triglyceride (r=?0.639, P<0.05). These parameters should be considered when developing procedures for either artificial fertilization or for cryopreservation of grass carp sperm.     

灌喂必需氨基酸模式溶液对草鱼全鱼_省略_肌肉_肝胰脏蛋白质合成代谢的影响

采用大剂量食道灌喂DL－[4-^3H]-Phe，研究不同EAA模式下，20－30g体重草鱼种的全鱼和肌肉，肝胰脏的蛋白质合成代谢，结果表明：（1）分别缺乏Lys,Met,Trp Arg的4种极端不平衡EAA模式，同草鱼肌肉EAA模式即相对平衡的基础模式相比，肌肉，肝胰脏及全鱼的蛋白质合成速率（FSR）均显著降低，其降低程度高低依次为Met,Lys,Arg,Trp;(2)肝胰脏，肌肉人的蛋白质合成代谢与整体蛋白质合成代谢对EAA模式的改变产生相同的变化趋势，并且三者蛋白质合成代谢速率为肝胰脏＞全鱼＞肌肉；（3）EAA模式的Lys,Met,Trp,Arg4因素中，对肌肉，肝胰脏蛋白质合成速率影响最大的是Met,其余依次为Lys,Arg,Trp。     

草鱼–鳙–鲫零换水池塘有机碳、氮、磷收支研究

为研究草鱼(Ctenopharyngodon idella)–鳙(Aristichthys nobilis)–鲫(Carassius carassius)零换水池塘营养盐收支状况,阐明其零换水机制,以草鱼–鳙–鲫零换水池塘为实验组,以草鱼–鳙–鲫常规换水池塘为对照组,开展了为期2年的池塘有机碳(TOC)、氮(N)、磷(P)收支的研究。结果显示,2组池塘TOC、N、P的主要来源均为饲料投入,分别为77.06%和81.00%,92.08%和92.77%,94.18%和95.63%;TOC、N、P的主要输出途径均为底泥积累,分别占输入营养盐的43.32%和22.10%,61.40%和52.82%,78.71%和79.58%。2组池塘养殖鱼类收获分别占输入碳(C)、N、P的10.08%和13.05%,21.00%和25.57%,15.41%和18.60%。零换水池塘的C、N、P水体积累量和积累率均显著低于常规池塘(P<0.05),其积累率分别降低92.91%、88.52%和87.12%。零换水池塘的N、P底泥积累量显著高于常规池塘,但C、N底泥积累率显著低于常规池塘(P<0.05),分别降低了48.99%和13.97%。零换水池塘养殖鱼类的C、N、P利用率均显著高于常规池塘(P<0.05),分别提高了29.49%、21.72%和20.65%。研究表明,零换水模式能降低营养盐积累,有效提高系统物质利用率,是一种绿色高效养殖模式,具有较好的推广价值。     

在草鱼免疫反应中莨菪佐剂性的初步观察

在草鱼免疫反应中莨菪佐剂性的初步观察赵建培（福州市郊区水产技术推广站，福州３５００１１）史维舟，陈昌福（华中农业大学水产学院，武昌４３００７０）已有研究报道指出，英若具有增强鱼类免疫力的作用［１］。但是，关于度著究竟是如何作用于鱼类的免疫系统，即为什...     

草鱼种对配合饵料中钙磷的需要量及适宜钙磷化

研究了草鱼种对饵料中钙磷的需要量及适宜钙磷比。试验结果以增重率和饵料转换率两项生物学指标为依据进行综合分析。结果表明，在本试验条件下草鱼对饵料中钙的需要量为0.473%～0.788%，磷的需要量为1.419%～1.577%，钙磷的总需要量在2.100%～2.700%范围内，适宜钙磷比则随着钙磷含量的高低同异而变化，即在钙磷含量较低而总量不同时为1:3,钙磷含量而总量相同时为1:2。当钙磷比中的磷小于2时，钙磷总量高则可减轻因钙磷不平衡对草鱼生长所产生的抑制作用，钙磷比不等于1:2(或1:3）时，钙磷总量过多或过少都不利于鱼的生长、且饵料转换率呈降低趋势。     

穿心莲对草鱼肠内细菌的影响

给不同组草鱼分别投喂含0％、0.5％、1％和2％穿心莲的人工配合饲料，在不同时期取样检测各组草鱼前、中、后肠需氧和兼性厌氧菌的种类和数量。结果显示，草鱼肠内细菌的种类，特别是优势菌群的种类没有受到穿心莲的影响。各组草鱼肠内各部位的优势菌群均为气单胞菌Aeromonas和肠杆菌Enterobacteriaceae。草鱼肠内细菌数量为105～108CFU·g-1，分析后发现，各组草鱼肠内细菌数量之间没有显著差异(P＞0.05)。但1％及2％的穿心莲能降低气单胞菌的组成。上述结果表明，一定浓度的穿心莲可以通过降低优势菌群气单胞菌的组成而对草鱼肠内微生态系产生影响。     

池塘中亚硝酸盐对草鱼种的毒害及防治

亚硝酸盐对草鱼种有很高的毒性,特别是水温超过30℃以上时。它主要是诱导草鱼血液中的血红蛋白转变为高铁血红蛋白,高铁血红蛋白超过血红蛋白总量的23.0％时,容易诱发草鱼出血病。随着草鱼血液中高铁血红蛋白的增高,红细胞数量和血红蛋白含量逐渐减少。池中亚硝酸盐氮含量到0.099±0.03毫克/升时,草鱼会出现出血病。清除水中亚硝酸盐氮的方法是在池塘中施加次氯酸钠,亚硝酸盐氮超过0.15毫克/升时,最好在池塘中先施加熟石灰,然后施加次氯酸钠,且以分次施用,效果较好。     

Grass carp (Ctenopharyngodon idella) grazing on duckweed (Spirodela polyrhiza)

The aims of this experiment were (1) toquantify the ability of grass carp to processduckweed and (2) to assess indirect changes inwater chemistry and phytoplankton community,caused by grass carp feeding. Yearling grass carp sized 126 ± 7.7 mm (TL) and19.6 g in weight were kept in 9 laminate tanksof 1 m³ for 14 days. Two stockingdensities (2 and 6 fish per m³) anda control without fish were used. Standard growthrate (SGR) of grass carp fed exclusively onduckweed was 0.70% body weight (BW) d^–1and food conversion ratio (FCR) reached 2.0(average water temperature =21.1 ± 3.8 °C). Daily food intakewas 0.2 g of duckweed dry weight (DW), i.e.,1% of average BW of grass carp. SGR ofduckweed growing in 20 × 20 cm floatingenclosures, differed significantly[F(6,2) = 417.9; p = 0.002] between the twostocking densities of grass carp and thecontrol tanks (without fish). Mean SGR ofduckweed was 0.02 g g^–1 day^–1 and thehighest SGR was recorded in the control tanks.Both decrease in NH₄-N and increase inNO₂-N concentrations differedsignificantly between the treatments[F(2,2) = 45.3; p = 0.02 and F(2,2) = 19.2; p = 0.04 respectively]. Changes in other nitrogenand phosphorus components (NO₃-N, TN, TPand PO₄-P) caused by stocking of grasscarp were not significant. Biomass ofphytoplankton, dominated by filamentous algaeand blue-greens, increased proportionately tostocking density of grass carp. Althoughduckweed has a large potential for nutrientremoval, the most common pathway for thenutrients released through grass carp grazingif duckweed cover is loose is theirincorporation into phytoplankton biomass.