团头鲂三倍体的微卫星遗传特征及生长性能分析

为研究团头鲂三倍体育种,对团头鲂三倍体和二倍体2个群体进行微卫星遗传结构特征分析和生长性能比较.利用筛选得到的20个微卫星标记对团头鲂三倍体和二倍体进行遗传多样性分析.试验结果表明,团头鲂三倍体、二倍体的平均等位基因数分别为2.75、3.05,平均期望杂合度分别为0.5061、0.5412,平均多态信息含量分别为0.4...     

二倍体与三倍体山女鳟的血液学比较分析

对二倍体和三倍体山女鳟(Oncorhynchus masou)的全血、血浆各种特征及结构进行血液学分析,结果表明,三倍体山女鳟的红细胞(RBC)体积与二倍体的比值为1.48±0.19.三倍体的红细胞数和红细胞压积(Hct)均显著低于二倍体,但其血红蛋白(Hb)浓度与二倍体的无显著性差异(P>0.05).这些血液生理学数据表明,二倍体与三倍体山女鳟可能具有相似的氧运输和有氧代谢能力.血液生化结果显示,二倍体和三倍体山女鳟的生化指标检测值不存在显著性差异(P>0.05);回归分析结果显示,与二倍体和三倍体山女鳟血浆参透压(Osm)相关性最大的生化指标均为Na+浓度、血糖浓度和尿素氮浓度.虽然三倍体与二倍体山女鳟血红蛋白含量之间不存在显著性差异,但由于三倍体的红细胞数远低于二倍体,加之三倍体的全血黏度较二倍体的高,在应激或低溶解氧的情况下,三倍体将无法动员更多的红细胞进行组织有氧代谢,因此,与二倍体相比,三倍体山女鳟的组织缺氧或窒息的风险更大,对溶解氧的要求更高.     

二倍体与三倍体虹鳟(Oncorhynchus mykiss)红细胞及部分血液指标的比较

对三倍体和二倍体虹鳟(Oncorhynchus mykiss)的红细胞大小(长、短径值和体积)、形态及部分血液指标(红细胞数、血红蛋白、红细胞压积、红细胞脆性和白细胞数)进行了比较。结果表明:三倍体虹鳟红细胞体积与二倍体比值为1.51∶1,细胞核大小与二倍体比值为1.49∶1,与理论值(1.5∶1)比较均无显著差异(P>0.05)。二倍体红细胞短径/长径比值为0.63,呈现出椭圆形;三倍体红细胞短径/长径比值为0.60,呈现出明显的长椭圆形,并有19%的红细胞有哑铃形或分裂的细胞核,这可作为区别二倍体鱼类的一个重要鉴定标志。三倍体虹鳟的红细胞数、红细胞压积和红细胞脆性明显小于二倍体(P<0.05),而血红蛋白与白细胞数与二倍体比较无明显差异(P>0.05)。     

二倍体、三倍体虹鳟的血液指标分析

通过对二倍体和三倍体虹鳟红细胞大小的测量,发现两者之间存在着极其显著的差异.三倍体虹鳟红细胞长径和核长径分别是二倍体的1.34和1.33倍;红细胞面积和核面积分别为二倍体的1.43和1.41倍:核体积为二倍体的1.47倍.用MEK-6318血液自动分析仪检测二倍体和三倍体虹鳟血常规值,结果显示:在红细胞计数(RBC)和红细胞平均容量(MCV)上,二、三倍体之间都有着极其显著的差异.此外,三倍体虹鳟的淋巴细胞、中间细胞和中性粒细胞数量都显著超过了二倍体.     

三倍体与二倍体泥鳅的组织细胞学比较

为探明三倍体和二倍体泥鳅的组织细胞学差异，运用血液涂片及组织切片方法，对三倍体、二倍体泥鳅的红细胞及其细胞核的长轴和短轴，血液中血细胞的组成及仔鱼眼部构造等进行了比较。结果表明，三倍体泥鳅红细胞及其核的长轴均比二倍体的要大，其中前者红细胞核的长轴约为后者的1．12倍，红细胞体积、核体积分别约为后者的1．62和1．44倍。三倍体泥鳅红细胞的相对数量明显少于二倍体泥鳅，而嗜中性细胞、单核细胞及淋巴细胞则多于后者。此外，三倍体泥鳅眼部色素层细胞数量少于二倍体。     

三倍体与二倍体泥鳅的组织细胞学比较

为探明三倍体和二倍体泥鳅的组织细胞学差异,运用血液涂片及组织切片方法,对三倍体、二倍体泥鳅的红细胞及其细胞核的长轴和短轴,血液中血细胞的组成及仔鱼眼部构造等进行了比较.结果表明,三倍体泥鳅红细胞及其核的长轴均比二倍体的要大,其中前者红细胞核的长轴约为后者的1.12倍,红细胞体积、核体积分别约为后者的1.62和1.44倍.三倍体泥鳅红细胞的相对数量明显少于二倍体泥鳅,而嗜中性细胞、单核细胞及淋巴细胞则多于后者.此外,三倍体泥鳅眼部色素层细胞数量少于二倍体.     

二、三倍体鲤鱼形态特征的比较研究

本文对二、三倍体鲤鱼的形态特征进行了较为全面的比较研究。结果表明：二、三倍体鲤鱼在体长／尾柄长、尾柄长/尾柄高、头长／尾柄长、头长／眼后头长、头长／眼间隔、体高／体厚等可量性状方面有显著性差异，据此可做为二、三倍体鲤鱼之间的区别特征。在可数性状方面没有明显     

熊本牡蛎三倍体与二倍体性腺发育特征比较

为了评估三倍体熊本牡蛎(Crassostrea sikamea)的繁殖潜力, 采用细胞松弛素 B 诱导了熊本牡蛎三倍体, 比较了 60 日龄(2016 年 8 月)~450 日龄(2017 年 9 月)三倍体与二倍体性腺发育特点, 分析了性腺发育与繁殖周期的相关性。研究结果表明, 熊本牡蛎三倍体与二倍体性腺发育均可分为形成期、增殖期、成熟期、排放期和耗尽期 5 个时期; 在一个繁殖周期内 22%的三倍体性腺可发育至成熟期, 但与二倍体相比, 三倍体成熟性腺的滤泡小、结缔组织丰富; 三倍体与二倍体性腺发育周期同步, 繁殖季节均位于 3—9 月, 繁殖期较长; 150 日龄(2016 年 10 月)三倍体与二倍体中发育的性腺(包括增殖期、成熟期、排放期、耗尽期)分别占 70%与 90%, 210 日龄(2017 年 1 月)减小至 3%与 18%, 之后性腺再次发育, 分别在 360 日龄(2017 年 6 月)和 450 日龄(2017 年 9 月)成熟期性腺比例达到最大值(40%和 90%)。三倍体与二倍体雌雄比分别为 1.35 : 1 和 0.95 : 1, 三倍体性比显著偏离 1 : 1 (P<0.01)。性腺成熟期三倍体与二倍体平均卵径分别为(44.30±6.88) μm 与(37.76±5.76) μm, 三倍体卵径比二倍体大 17.33% (P< 0.05)。本研究可为熊本牡蛎三倍体和二倍体繁育提供参考依据。     

三倍体虹鳟制种技术与倍性鉴定

虹鳟(Oncorhynchus mykiss)属鲑形目、鲑科,是重要的冷水性养殖鱼类,三倍体虹鳟以其不育或育性不良、肉质提升和生长周期缩短等特点得到养殖者的喜爱。尽管国内多地已经进行了三倍体虹鳟制种的研究,但效果并不理想,目前三倍体虹鳟苗种主要依靠国外进口。本文拟对三倍体人工诱导和倍性鉴定两个方面进行总结,以利于更有效地进行三倍体虹鳟规模制种。     

长牡蛎诱导三倍体与二倍体的养殖生物学比较研究

许多水产动物的三倍体在养殖生产中显示出很大的增产潜力 ,因此 ,采用染色体操作技术诱导三倍体的研究越来越受到人们的重视。长牡蛎( Crassostrea gigas)是我国沿海重要的经济贝类 ,长期以来 ,由于牡蛎繁殖季节大量排放精、卵 ,一是引起蛎肉瘪小充水 (成为透明状水蛎 ) ,致使肉质变劣而无法上市。二是雌性牡蛎大量排卵导致生理活性降低 ,出现大批死亡。近年来 ,笔者对长牡蛎三倍体的诱导和培育技术进行了深入、系统的研究 ,在三倍体生长发育、生殖腺组织学和超微结构变化、三倍体软体组织生化成分含量周年季节变化以及二倍体和三倍体的鉴…     

Production of interploid triploids by 4n × 2n blunt snout bream (Megalobrama amblycephala. Yih) and their first performance data

Positive and negative interploid triploids of blunt snout bream (Megalobrama amblycephala) were produced by reciprocal crossing autotetraploids with diploids. Fertilization and hatching rate of negative interploid triploid, 2n♀× 4n♂, at different reproductive ages was similar to that of control, but higher than that of positive interploid triploid, 4n♀× 2n♂. Additionally, the development rate of embryos of the positive interploid triploid, 4n♀× 2n♂, was slower than that of control and the negative interploid triploid, 2n♀× 4n♂. Gonad growth of the positive and negative interploid 3n was significantly (P<0.01) affected by parents, and spawning could not be induced both in females and males at the 2+ age. The negative interploid 3n (2n♀× 4n♂) had a significantly higher survival rate than the positive interploid 3n (4n♀× 2n♂), but close to the level of the control (P<0.05). The daily growth rate of the negative interploid 3n was similar to that of the good breed ‘Pujiang No. 1’ blunt snout bream (2n) during the early life stages.     

Dietary histidine requirement of juvenile blunt snout bream (Megalobrama amblycephala)

An eight‐week feeding trial was conducted to determine the dietary histidine requirement of juvenile blunt snout bream. The results showed that final body weight, weight gain rate and specific growth rate significantly increased with increasing dietary histidine levels up to 9.9 g/kg (p < .05) and decreased gradually thereafter, while feed conversion ratio showed a converse trend. The survival rate, condition factor, viscerosomatic and hepatosomatic index were not significantly affected by the graded dietary histidine levels (p > .05). About 9.9 g/kg dietary histidine level significantly improved whole‐body protein and lowered moisture content. The contents of plasma total protein, cholesterol, urea and triglyceride levels were not significantly affected by dietary histidine levels. About 9.9 g/kg dietary histidine level significantly upregulated target of rapamycin and insulin‐like growth factor mRNA levels (p < .05), and 12.1 g/kg and 14.2 g/kg dietary histidine levels significantly upregulated eukaryotic translation initiation factor 4E‐binding protein 2 and ribosomal protein S6 kinase‐polypeptide 1 mRNA levels (p < .05). Based on second‐degree polynomial regression analysis of weight gain rate, and specific growth rate against dietary histidine levels, the dietary histidine requirement for juvenile blunt snout bream was estimated to be 11.2 g/kg of diet, corresponding to 36.1 g/kg of dietary protein.     

湖北淤泥湖团头舫mtDNA限制性片段长度多态性的研究

用ＢａｍＨＩ、ＢｇｌＩ、ＣｌａＩ、ＥｃｏＲＩ、ＨｉｎｄⅢ、ＫｐｎＩ、ＰｓｔＩ，ＰｖｕⅡＳａｃⅡ、ＳａｃＩ、ＳｃａＩ、ＸｂａＩ，ＸｂａＩ和ＸｈｏＩ十四种限制性内切酶对来源于湖北淤泥湖的团头舫进行线粒体ＤＮＡ限制性片估长度多态性研究。初步表明ＤＮＡ上切点呈现不同程度的多态性。共检测到六种母集集团。发现一尾团头舫ｍｔＤＮＡ分子大小比普通型鱼ｍｔＤＮＡ小约０．７０ｋｂ。呈现出ｍｔＤＮＡ发子大小多态现象。     

果寡糖对急性氨氮应激下团头鲂非特异性免疫指标的影响

为研究氨氮应激条件下果寡糖对团头鲂(Megalobrama amblycephala)非特异免疫指标的影响,试验选择360尾初重为13.5 g的团头鲂随机分为3组,每组4个重复,每缸30尾鱼,对照组投喂基础日粮,试验组分别投喂果寡糖(FOS)添加量为0.4%和0.8%的日粮,饲养8周,养殖试验结束后,用10 mg/L的NH4Cl对24尾鱼进行氨氮应激试验。试验结果显示,氨氮应激前,0.4%FOS组的血液谷丙转氨酶(GPT)活性显著低于对照组和0.8%FOS组,在氨氮应激后,0.4%FOS组的GOT活性有降低的趋势,但仅在24 h时显著低于0.8%组,0.8%组和对照组之间并无显著差异;血清酸性磷酸酶、补体3(C3)、补体4(C4)、酚氧化酶都出现先升高后降低的趋势,分别在应激后的3、24、12、12 h达到最大值。此外,应激前0.4%FOS组补体C3、C4、酚氧化酶的活性都显著高于对照组,应激后0.4%FOS组C3在3 h处显著高于对照组,但是在其它时间点各组差异并不显著,果寡糖添加水平和采样时间对这些免疫指标都无显著的交互作用。结果表明饲料中添加0.4%的果寡糖能够提高团头鲂的免疫力指标活性,提高了团头鲂抗氨氮应激的能力。     

团头鲂3个选育群体遗传潜力的微卫星分析

为从遗传多样性的角度了解团头鲂(Megalobrama amblycephala)3个选育群体的遗传潜力,该研究以团头鲂"浦江1号"选育奠基群体(F_0)为对照组,采用14个多态性转录组微卫星标记评估了团头鲂3个选育群体的遗传多样性,分析其遗传潜力。结果显示,3个选育群体平均每个位点的等位基因数(A)为7.928 6~8.785 7,有效等位基因数(A_E)为4.409 4~4.878 4,观察杂合度(H_O)为0.491 1~0.574 4,期望杂合度(HE)为0.741 3~0.751 8,多态信息含量(PIC)为0.691 2~0.705 2,近交系数(FIS)为0.229~0.352。3个选育群体的遗传多样性水平(AE、HE)均高于F0群体,但不存在显著差异(P0.05)。3个选育群体的有效群体大小(N_e)为11.0~29.3,在近期可能经历过遗传瓶颈。3个选育群体间D_A、D_(SW)遗传距离分别为0.175 4~0.358 8、0.804 7~1.054 4。该结果表明,3个选育群体的遗传多样性较高,遗传潜力较大,但因有效群体数量较少和瓶颈效应的影响,存在杂合度下降和近交衰退的风险,今后需采取科学措施来保护选育群体的遗传潜力。     

Dietary choline requirement for juvenile blunt snout bream,Megalobrama amblycephala

A 12‐week feeding trial was conducted to determine dietary choline requirement for juvenile Megalobrama amblycephala. The basal diet was formulated to contain 310 g kg^?1 diet from vitamin‐free casein and gelatine. Choline chloride was supplemented to the basal diet to formulate six purified diets containing 0, 250, 500, 1000, 2000 and 4000 mg kg^?1, respectively. Each diet was randomly fed to quadrupled groups of Megalobrama amblycephala with initial average weight 1.84 ± 0.04 g in a flow‐through system. Results showed weight gain was increased significantly with increasing dietary choline levels (P < 0.01). Lipid content of liver decreased significantly as dietary choline concentration increased (P < 0.01), whereas lipid content of dressed carcass showed opposite trend (P < 0.01), and lipid content of whole‐body was unaffected by dietary choline supplementation. Broken‐ line regression of weight gain, liver and muscle choline concentration showed choline requirements of Megalobrama amblycephala of 1198, 1525 and 1365 mg kg^?1, respectively. In addition, dietary choline supplementation significantly improved lipid content of dressed carcass but not the content of whole body of blunt snout bream.     

Dietary methionine requirement of juvenile blunt snout bream (Megalobrama amblycephala) at a constant dietary cystine level

A 9‐week feeding trial was conducted to investigate the dietary methionine requirement of juvenile Megalobrama amblycephala at a constant dietary cystine level. Six semipurified diets were formulated to contain graded dietary methionine levels from 3.9 to 15.4 g kg^?1 in about 2.5 g kg^?1 increments in the presence of 2.2 g kg^?1 cystine. Results showed that specific growth rate (SGR) and protein efficiency ratio (PER) significantly increased with increasing dietary methionine levels from 3.9 to 12.4 g kg^?1 and thereafter kept stable. Maximum protein productive value (PPV), nitrogen retention efficiency (NRE) and liver weight were observed in 8.5 g methionine kg^?1 diet. Protein contents in whole fish body were positively correlated with dietary methionine level, while lipid contents were negatively correlated with it. Morphological index and hepatic glutamate‐pyruvate transaminase (GPT) activities were independent of dietary methionine levels. However, dietary methionine supplementation significantly improved haematological parameters, plasma methionine and total essential amino acid contents and hepatic glutamate‐oxaloacetate transaminase (GOT) activities. Analysis of dose response using broken‐line regression on the basis of SGR and PPV versus dietary methionine level estimated the optimum dietary methionine requirements of juvenile M. amblycephala to be between 8.5 and 8.4 g kg^?1 of diet (25.0 and 24.7 g kg^?1 of protein) in the presence of 2.2 g kg^?1 cystine, respectively. Hence, the corresponding total sulphur amino acids requirements of this species were calculated to be 10.7 and 10.6 g kg^?1 of diet (31.5 and 31.2 g kg^?1 of dietary protein).     

Dietary isoleucine requirement of juvenile blunt snout bream,Megalobrama amblycephala

A 9‐week feeding trial was conducted to estimate the dietary isoleucine requirement of juvenile blunt snout bream. Six isonitrogenous and isoenergetic experimental diets were formulated to contain graded isoleucine levels ranging from 5.3 to 20.1 g kg^?1 dry diet. At the end of the experiment, weight gain (WG), specific growth rate (SGR), feed efficiency ratio (FER) and protein efficiency ratio (PER) increased with increasing dietary isoleucine level up to 11.1 g kg^?1 dry diet, and dietary isoleucine level above 14.2 g kg^?1 dry diet declined these performances. Dietary isoleucine levels (14.2 and 17.3 g kg^?1 dry diet) significantly improved whole‐body protein content, but decreased whole‐body lipid, plasma triglyceride and cholesterol contents. Significantly lower visceral fat index (VFI) in fish fed with 14.2 g kg^?1 dietary isoleucine was observed compared to those fed with deficient or excessive isoleucine. Dietary isoleucine supplementation significantly increased plasma isoleucine concentration, while plasma valine and leucine concentrations showed a reversed trend. Dietary isoleucine levels regulated the target of rapamycin (TOR) gene expression and improved plasma superoxide dismutase (SOD) activity in juvenile blunt snout bream. Based on second‐order polynomial regression model analysis of SGR and FER, the optimum dietary isoleucine requirement was estimated to be 13.8 g kg^?1 dry diet (40.6 g kg^?1 dietary protein) and 14.0 g kg^?1 dry diet (41.2 g kg^?1 dietary protein), respectively.     

团头鲂嗅觉器官的发育及代表性嗅觉受体基因的表达模式研究

为了研究团头鲂嗅觉器官的发育过程和嗅觉受体基因（Olfactory receptor genes,ORs）在嗅囊发育不同时期的组织表达模式,本实验基于组织学、形态学及qRT-PCR的方法进行了探究。组织形态学结果显示团头鲂嗅觉器官位于后背部和两侧眼睛前方,嗅囊紧贴于嗅腔底部,形状近似椭圆形,嗅基板沿着头尾方向平行排列。嗅囊在团头鲂刚孵化时出现嗅窝外围凹进的痕迹,随后凹进处长出嗅窝,并在中央出现长条形的嗅基板。随着仔鱼的生长发育,单侧嗅囊初级嗅基板排列由疏松逐渐紧密,嗅基板隆起高度、数量及嗅囊总体表面积逐渐增加,在成鱼阶段趋于稳定;qRT-PCR结果显示代表性ORs在团头鲂发育不同时期的组织中表达模式各异,其中Beta-2、9、10、11在胚胎发育期的囊胚期、胚孔闭合期及嗅板期高表达（P<0.01）;Beta-2、9、10、11和Epsilon-7在仔稚鱼期的3至15 dpf中微弱表达,在30至60 dpf时期高表达（P<0.01）;Beta-2、10、11及Epsilon-6、7、10、13在幼鱼期至成鱼期嗅囊中高表达（P<0.05）,Beta-2、10、9、11及Epsilon-7、10、13在3月龄团头鲂嗅球中高表达（P<0.01）;Beta-2、10在幼鱼期至成鱼期脑中低表达（P<0.05）。总之,该研究系统探究了团头鲂嗅觉器官的发育进程与ORs表达规律,为后续深入研究鱼类嗅觉识别机制奠定基础。     

团头鲂野生、驯养、选育3类遗传生态群体遗传变异的线粒体DNA分析

通过对线粒体DNA控制区和COⅠ基因序列的联合分析,研究了团头鲂(Megalobrama amblycephala)3类遗传生态群体(包含4个野生群体、2个驯养群体、1个选育良种"浦江1号"群体)的遗传多样性和遗传分化情况.结果表明:(1)在所分析的7个群体中,共确定了64种单倍型,群体间无共享单倍型.(2)4个野生群体内线粒体DNA的单倍型多样度(Hd)在0.857～0.943之间,核苷酸变异位点数在31～40之间,核苷酸多样性指数(π)在0.275%～0.461%之间,平均核苷酸差异数(K)的范围为4.043～6.800;2个驯养群体的相应参数变化范围分别为0.714～0.800、18～21、0.122%～0.175%、1.800～2.586,均低于野生群体;选育群体的相应参数分别为0.843、23、0.193%、2.843,低于4个野生群体,但高于2个驯养群体.以上4种多样性参数在7个群体中的变化趋势一致.(3)7个群体之间的平均遗传距离在0.000 6～0.003 5之间,遗传分化指数(FST)在0.010 9～0.133 1之间.4个野生群体间FST值差异不显著(P＞0.05),而2个驯养群体间FST值差异显著(P＜0.05),它们与选育群体间的FST值差异也显著(P＜0.05).以上结果表明,生存环境的殊异(敞开的天然水体,封闭的驯养池塘)和人工选择(严格有序的科学选育)对种群遗传结构影响巨大,导致鱼类不同遗传生态类型群体间产生遗传变异和遗传分化.