条斑星鲽精巢年周期发育规律和血液性类固醇激素含量变化

采用组织学方法和放射免疫法(RIA)等技术方法,研究了人工养殖条件下条斑星鲽雄性亲鱼精巢发育规律和性类固醇激素的年周期变化规律。实验结果表明,条斑星鲽精巢中可见5个时相的生殖细胞类型,精巢发育可分为5期。条斑星鲽雄鱼GSI值自10月开始升高,12月达峰值(P≤0.05),之后显著下降并保持较低值至下一次生殖周期开始。HSI值在11～12月保持较高表达值,其他各月份保持相对稳定水平。CF值在5～8月保持较高水平,其他月份保持平稳状态。雄鱼血浆中睾酮(T)水平自9月开始升高并在12月(Ⅴ期精巢)达到峰值,其后显著下降并在其后的月份保持较低水平,而雌二醇(E2)在2月出现峰值,其他月份保持相对稳定水平。     

养殖圆斑星鲽血浆性类固醇激素表达与卵巢发育及温光调控的关系

系统研究了人工养殖条件下圆斑星鲽(Verasper variegatus)亲鱼血浆性类固醇激素的年周期变化规律及其与卵巢发育成熟及温光调控的关系.结果表明,圆斑星鲽卵巢发育属非同步分批发育模式,一年一次成熟分批产卵类型.卵巢年周期发育过程中可见5个发育时相的卵母细胞,卵巢发育可化分为4个成熟阶段,6个发育时期.性腺指数(GSI)、肝脏指数(HSI)和肥满度(CF)在亲鱼的年周期发育中呈现规律性的变化.亲鱼血浆中雌二醇(E2)在产卵期达峰值;排卵结束后,E2表达水平降至较低水平.血浆中睾酮(T)含量在10月达峰值,并在产卵期间保持相对较高表达水平.统计分析表明,血浆E2水平与GSI和HSI值的变化都呈现显著的正相关关系（P＜0.05）.水温和光周期对卵巢发育具有明显的影响并显著影响E2的表达,GSI与水温存在显著的负相关关系(P＜0.05）,HSI与水温存在显著的负相关关系(P＜0.05),而CF与光周期现显著的负相关关系(P＜0.05).结论认为,圆斑星鲽雌性亲鱼血浆性类固醇激素表达水平变化与性腺发育（卵母细胞成熟）、水等具有显著的相关关系.本研究结果可作为温光调控人工亲鱼性腺发育成熟、激素诱导亲鱼产卵的重要参考技术依据.     

养殖银鲳性腺发育规律和性类固醇激素变化

为掌握养殖银鲳的性腺发育规律及其与性类固醇激素水平的关系,本研究采用组织切片技术和酶联免疫吸附法(Elisa),观察养殖条件下银鲳的精巢与卵巢发育特征和血清中性类固醇激素的周年变化,并分析性腺成熟指数(GSI)、肝重指数(HSI)和肥满度(CF)与性腺发育的关系。结果显示,养殖银鲳雌鱼1龄即可发育成熟,成熟卵巢呈一对""形的囊状结构。11月份有近一半个体发育至III期,33.3%已发育至IV期,12月—翌年1月越冬期间约2/3退化至II期,3月又迅速发育至IV期,4月份达到V期并产卵,5月份产卵结束卵巢进入恢复期IV期。银鲳精巢属小叶型,III期精巢出现小叶腔,IV期精巢出现精子,精巢发育速率快于卵巢,即"雄性早熟",7月龄约20%个体可达到性成熟,越冬时退化至III期,3月重新发育,精子分批成熟排精。雌鱼GSI值为0.19%~12.89%,HSI为0.97%~2.30%,CF为2.30~3.08 g/cm~3,雄鱼的GSI为0.08%~2.62%,HSI值为0.73%~1.83%,CF为2.11~2.80 g/cm~3,GSI值在V期时达到最大,HSI和CF值则于IV期达到峰值。雌鱼血清中雌二醇(E2)周年表达水平为23.27~59.13 pg/m L,雄鱼为15.90~36.20 pg/m L,雌鱼的睾酮(T)为14.57~68.67 nmol/L,雄鱼的T为18.62~66.49 nmol/L,E2水平与HSI值呈现显著正相关关系,T表达水平与GSI值呈现显著正相关关系。研究表明,银鲳血清中性类固醇激素含量与精巢和卵巢的发育密切相关,可作为了解养殖银鲳性腺发育的重要指标。     

圆斑星鲽卵巢发育的组织学和数量形态特征研究

研究了圆斑星鲽卵巢年周期发育的组织学规律及其数量形态特征变化.结果表明,圆斑星鲽产卵属1年1次成熟分批产卵类型,卵巢发育属非同步分批成熟类型.性成熟亲鱼在年周期发育中经历5个发育时期,同时可见5个发育时相的卵母细胞,不同发育时期的卵巢中卵母细胞类型和组成比例不同.性腺指数(GSD、肝脏指数(HSI)和肥满度(CF)在亲鱼的年周期发育中呈现规律性的变化,与卵巢发育呈现密切的关联.亲鱼卵巢发育的有效积温为1050日度.     

条斑星鲽人工繁育技术研究

对条斑星鲽人工繁育技术,孵化适温、适盐范围及亲鱼、仔稚鱼营养强化进行了研究,探讨了温度、盐度、营养强化与性腺、胚胎、胚后发育的关系。结果表明:采用先降温再升温+营养强化方法促熟亲鱼可获得优质卵子。最适孵化温度(10.5±0.5)℃,最适孵化盐度33.6。以小球藻强化轮虫有助于提高仔鱼开口率、生长率及成活率。     

条斑星鲽精子超微结构研究

分别以扫描电镜和透射电镜方法研究了条斑星鲽精子的超微结构。条斑星鲽精子全长45.57±3.23μm,主要包括三部分:头部、中段和鞭毛。头部呈圆形,主要结构为近圆形的细胞核;中段不发达,主要包括线粒体和中心粒复合体两部分,具有多个未分化的圆形小线粒体;鞭毛细长,结构为典型的"9+2"型双联微管结构,无侧鳍。这些特征表明,条斑星鲽的精子属于简单的原始类型。条斑星鲽精子不具顶体,但具有开口于核前端稍偏处呈不规则状的核前部凹陷及核内核泡结构,核前凹陷与染色质区无界膜分隔,部分精子在凹陷前端存在由核膜构成的凸起。凹陷区的前部及其开口处常有多个小的具单层界膜的囊泡,这些结构可能是顶体的遗迹,但也不排除其是向外释放核内物质的一种途径的可能性。     

条斑星鲽染色体带型研究

开展了条斑星鲽染色体C-带、Ag-NORs带及G-带的研究.结果发现,条斑星鲽染色体显示较为复杂的C-带,有4对染色体整体呈阳性深染,分别为3、13、20、22号染色体;3对具居间C-带,分别为1、4、8号染色体;9号染色体整体显示阴性,其余有大小不一的着丝粒C-带.计算其异染色质含量为54.91％.银染分裂相中Ag-...     

条斑星鲽(Verasper moseri)工厂化养殖技术

本文系统地介绍了条斑星鲽工厂化养殖的主要技术。从养殖设施、苗种的选择和运输、养殖管理、病害的防治、适时起捕上市方面进行了详细介绍,一般放养全长5cm的苗种,养殖10个月体重可达到600～800g,养殖16个月体重可达1200g,养殖前景广阔,具有很高的经济价值与养殖价值,是一个很值得推广的优良品种。     

条斑星鲽染色体带型研究

开展了条斑星鲽染色体C-带、Ag-NORs带及G-带的研究。结果发现,条斑星鲽染色体显示较为复杂的C-带,有4对染色体整体呈阳性深染,分别为3、13、20、22号染色体;3对具居间C-带,分别为1、4、8号染色体;9号染色体整体显示阴性,其余有大小不一的着丝粒C-带。计算其异染色质含量为54.91%。银染分裂相中Ag-NORs出现的数目以两个的频率最高,达87.5%,与此相一致,在间期核中通过银染表现出两个核仁的间期核数目的频率也最高,故确定条斑星鲽染色体具有1对Ag-NORs,位于亚中部着丝点染色体的短臂上,为端部Ag-NOR。Ag-NORs的数目为1~3个,以出现两个Ag-NOR的频率最高,达87.5%,与此相同的是,在间期核中,通过银染表现出两个核仁的间期核数目的频率也最高。G-带研究显示,条斑星鲽有37条深染带,22条浅染带。     

雌核发育与正常二倍体条斑星鲽胚胎发育比较研究

在用牙鲆精子诱导条斑星鲽雌核发育的同时,对条斑星鲽单倍体、雌核发育胚胎发育过程进行了观察,并与普通胚胎发育作了比较.观察结果表明,(1)单倍体胚胎,卵裂开始至64细胞期,发育速度较普通胚胎发育发育速度明显缓慢,至多细胞期发育速度相同,历时14 h;到达高囊胚期比普通二倍体慢1.5 h,到达低囊胚期发育速度相同,历时31 h;到达原肠末期、神经胚期、晶体形成期历时相同,其他发育时期一直处于缓慢状态;孵化仔鱼具有明显的单倍体综合症.(2)雌核发育与普通胚胎的发育的特征相比无明显区别,在卵裂期发育速度明显迟缓,但在囊胚期以后,所用时间基本相同;孵化时间比普通胚胎长约1 h.(3)与普通胚胎相比,雌核发育胚胎、单倍体胚胎孵化率较低,仔鱼畸形率高.     

Aquaculture performance of triploid barfin flounder Verasper moseri

ABSTRACT:   To evaluate the aquaculture performance of triploid barfin flounder Verasper moseri , the sex ratio, maturation, growth and the relative proportion of body parts were examined. The sex ratio of triploids was similar to diploids under communal rearing conditions, but the proportion of female diploids was higher than that of triploids under separate rearing conditions. The gonadosomatic index of triploid females was very low even during the spawning season, and the ovaries were rudimentary. These results suggest that triploid barfin flounder females were sterile. In addition, triploid males produced a small quantity of milt containing very few spermatozoa with abnormal shapes. Spermatozoa obtained from triploids were aneuploidies. When normal eggs were fertilized with sperm from triploid males, no fry developed. These results suggest that triploid barfin flounder males were functionally sterile. Triploid males grew more slowly than diploid males, and triploid females showed similar or slower growth than diploid females, whether reared separately (23 months) or communally (35 months). The ratios of visceral weight to the edible parts for triploid males were similar to those for diploid males, but ratios for triploid females were higher than for diploid females during the spawning period. In conclusion, a significant improvement of growth was not found in triploid barfin flounders.     

温度、盐度对条斑星鲽胚胎发育的影响

就温度、盐度对条斑星鲽胚胎发育的影响作了初步探讨。实验表明,胚胎发育的适宜温度为6～12℃,最适水温为8～10℃,在适温范围内胚胎孵化速度与温度呈正相关关系（N=65.925T^-0.9672）。盐度低于27,受精卵下沉,27～30之间悬浮于水体中,盐度高于35,受精卵浮于水面。胚胎孵化的适宜盐度范围、最适盐度范围分别为25～40和28～36;盐度低于25或高于40,孵化率都降低。研究表明,条斑星鲽胚胎发育的生物学零度（阈温度）为-0.1℃,有效积温值为1726.48℃.h。另外,根据其Q10值推算的胚胎发育的适宜水温与实验结果相似。     

Stress inhibits insulin release induced by feeding and arginine injection in barfin flounder Verasper moseri

We evaluated the effect of stress and adrenaline injection on insulin release in barfin flounder Verasper moseri. Flounders were fed to satiation and then exposed to 30 min of stress (chasing using a blunt stick), beginning 15 min after feeding. Plasma insulin concentrations were measured using a time-resolved fluoroimmunoassay for barfin flounder insulin. Peak insulin concentrations in the stressed flounders (3.4 ± 1.0 ng/ml, mean ± SEM) were lower than in the control group (8.3 ± 1.7 ng/ml). Similarly, in flounders injected with l-arginine (Arg; 3.5 mmol/kg body weight), the most potent insulinotropic factor for flounders, insulin levels were lower for up to 5 h in individuals exposed to chasing stress. Furthermore, plasma insulin levels did not increase after injection with Arg followed by exposure to 10 min of stress. This was also true for flounders co-injected with Arg (20 nmol/kg body weight) and adrenalin instead of the chasing stress. We conclude that stress induces a powerful and extended inhibition of insulin release in flounders, even when the stress is acute. We hypothesize that this effect is mediated partly by adrenaline release. Our results provide insight into the deleterious effects of stress on growth.     

条斑星鲽的工厂化养殖技术

近年来,随着我国北方海水鱼类工厂化养殖的发展,海水鱼类的养殖形成了一个规模化的现代养殖业。由于大菱鲆市场价格的大幅下滑,在我国北方以半滑舌鳎和圆斑星鲽为主要开发和研究对象的同时,条斑星鲽以其外观漂亮优美,体色似松树皮,富含胶原蛋白,肉质细嫩鲜美,深受消费者喜爱,因生长快,经济价值高,成为研究人员和海水养殖业主要研究和开发的对象之一,具有很大的开发价值和养殖潜力。笔者从日本引进条斑星鲽苗种2000尾,经过2年的养殖,平均体重达2.3kg,成活率95%以上。试验表明,条斑星鲽是工厂化养殖的优良开发品种。     

Gonadal maturation and spawning migration of stocked female barfin flounder Verasper moseri off the Pacific coast of northern Japan

Commercial catches of barfin flounder Verasper moseri, an important target flatfish for stock enhancement programs in northern Japan, have recovered remarkably since the large-scale release of seedlings from Hokkaido in 2006. However, their reproductive ecology remains unclear. Our study of 2008–2012 investigated seasonal changes in the maturity of stocked female barfin flounder (n = 4,123) off the Pacific coast from Hokkaido to Tohoku (35.7°–43.4°N). Histological observation of ovaries revealed that fish undergoing vitellogenesis were observed mainly off Hokkaido and partially off northern Tohoku during August–January (40.2°–43.4°N). However, fully matured or spawning fish were not found near those areas. They were observed only in southern Tohoku. Ongoing spawning was observed off southernmost Tohoku (35.8°–37.5°N around 300 m depth) from early February through late April, when commercial landings had decreased drastically off Hokkaido. Spent fish were caught only rarely off southern Tohoku, but were observed frequently off Hokkaido during April–June. These results demonstrate that spawning grounds form at the upper continental slope off southernmost Tohoku during February–April. Relations between maturation traits and seasonal landings strongly suggest that female barfin flounder repeatedly migrate more than 700 km from feeding grounds off Hokkaido to spawning grounds off southern Tohoku.     

Genetic evaluation of interindividual relatedness for broodstock management of the rare species barfin flounder Verasper moseri using microsatellite DNA markers

ABSTRACT:   The barfin flounder has been recently declared to be rare, as the number of wild individuals in Japan has greatly reduced since 1975. The Japanese National Center for Stock Enhancement started the stocking program for this species on 1987. In the present study, microsatellite DNA loci were applied in order to determine the pedigree of the hatchery-produced juveniles for stock enhancement. Additionally, using six polymorphic microsatellite markers, the accuracy to assign dyads to categories of kinship was estimated using the relatedness estimator ( R _XY), similarity indexes (individual genetic identity [GI] and proportion of shared alleles [ P _S]), identification of kin groups using unweighted pair group method with arithmetic mean (UPGMA) clustering based on interindividual genetic distances (1- R _XY, 1- P _S, and 1-GI), a likelihood ratio approach, and partition methods to separate individuals into cohorts. The results were compared with the pedigree relationship previously obtained from parentage analysis. Estimation of the kinship relationship between individuals could be better inferred by means of UPGMA dendrogram clustering based on 1-GI or 1- P _S as interindividual genetic distances, or using computer software to perform partitioning of set of individuals.