Assessing sexual maturity of feminized Japanese eel Anguilla japonica by measuring eye size

For efficient production of Japanese eel Anguilla japonica eggs, knowledge of the status of the sexual maturity of potential broodstock females is important because this status directly influences the time required to produce mature eggs by successive hormone doses. Here we apply an eye index (relative eye size) to evaluate the gonadal status of feminized A. japonica, which were induced by administration of estradiol-17β. Examination of gonad somatic and eye indexes of 267 feminized eels, cultivated for 12–56 months, revealed a significant correlation between these two indexes. Artificial maturation of 35 feminized eels showed that the number of hormone injections administered before ovulation was significantly lower as the eye index score increased, indicating availability of this noninvasive method of predicting sexual maturity of feminized eel. There was no correlation between eye index and fertilization rate, hatching rate, or larval survival rate, suggesting that sexual maturity before hormonal treatment does not affect egg quality.     

日本鳗鲡初孵仔鱼耗氧率的初步研究

研究了不同盐度、温度、光照条件下日本鳗鲡仔鱼(4.42±0.034mm)的耗氧情况。结果表明，幼鱼的耗氧率随盐度的升高而下降，随温度的升高而上升，耗氧率遮光比在自然光下要低。同时还测定了幼鱼的窒息点。     

Long-term rearing of Japanese eel larvae using a liquid-type diet: food intake,survival and growth

Fisheries Science - The efficiency of a new liquid-type diet for long-term rearing of Japanese eel larvae until metamorphosis was examined, as was the effect of diet viscosity on diet intake and on...     

中国近海日本鳗鲡玻璃鳗体组织营养成分及氨基酸、脂肪酸组成的比较

为了解日本鳗鲡玻璃鳗的早期营养需求,分别对中国近海6个不同地点玻璃鳗体组织一般营养成分、氨基酸和脂肪酸含量进行了分析。结果显示,野生玻璃鳗体组织中水分含量为78.62%±2.03%,干重基础下粗蛋白质含量为69.08%±0.64%、粗脂肪含量为11.96%±1.33%、粗灰分含量为11.45%±1.30%。在台州、舟山、南通和东台4个采样点的样品中,水分含量无显著差异,而粗蛋白含量以舟山点最高、粗脂肪含量最低,并且舟山点样品肠道内发现可见内容物。玻璃鳗样品中氨基酸含量以谷氨酸、天门冬氨酸、赖氨酸、亮氨酸最高,半胱氨酸最低;除舟山点外,各采样点氨基酸含量无明显差异。不同洄游地点玻璃鳗组织中脂肪酸含量均以C16:0、 C18:1n9、 C20:5n3(EPA)、 C22:5n3、 C22:6n3(DHA)为主要成分,其中EPA和DHA共占脂肪酸总量的31.59%±1.00%,不同洄游地点样品必需脂肪酸含量差异明显。研究表明,必需氨基酸、必需脂肪酸对于玻璃鳗的正常生长存活具有重要意义。此外,对比不同地点营养物质组成及稚鱼饵料理论需求量分析,推测舟山点海洋环境中包含更适合玻璃鳗稚鱼摄食的食物,...     

人工诱导日本鳗鲡性腺发育组织学研究

从人工诱导日本鳗鲡(Anguill japonica)性腺成熟的角度,采用对亲鳗群体外形和解剖观察,以及性腺组织切片的方法,对日本鳗鲡从下海鳗到性腺发育成熟的全过程进行了组织学和细胞学研究。结果表明,人工诱导的日本鳗鲡精巢发育仍可分为5个时相,即精原细胞前增殖期(Ⅰ);精原细胞后增殖期(Ⅱ),精母细胞生长期(Ⅲ),精子出现期(Ⅳ)和精子成熟期(Ⅴ)。除日本鳗鲡下海鳗的精巢有73.3%的个体处在第Ⅰ时相外,其整个精巢的发育过程和细胞学特征与以前的研究无太大差异。在下海鳗群体中未观察到第Ⅰ时相的卵巢。因此,人工诱导日本鳗鲡的卵巢发育只有卵母细胞单层滤泡期(Ⅱ);脂肪泡出现期(Ⅲ);卵黄充满期(Ⅳ)和卵母细胞最后成熟期(Ⅴ)。本研究除对人工诱导下日本鳗鲡性腺发育各时相的组织学和细胞学特征进行了观察外,还探讨了人工诱导日本鳗鲡性腺发育的时间、下海鳗群体的组成、以及脂肪泡的形成和变化过程等问题,为以后的日本鳗鲡人工育苗技术提供了参考依据。     

Expression of ubiquitin C-terminal hydrolase is regulated by estradiol-17β in testis and brain of the Japanese common goby

The effect of endocrine-disrupting chemicals (EDCs) on the expression of ubiquitin C-terminal hydrolase (UCH) mRNA in the Japanese common goby, Acanthogobius flavimanus, was investigated by competitive PCR assay. Exposure of fish to estradiol-17β (E₂) induced significant increase in the UCH mRNA in testis and brain, but exposure to bisphenol A and nonylphenol did not. This result suggests that expression of the UCH gene is E₂-inducible.     

日本鳗li初孵仔鱼耗氧率的初步研究

研究了不同盐度、温度、光照条件下日本鳗鲡仔鱼 ( 4 .4 2± 0 .0 34 mm )的耗氧情况。结果表明 ,幼鱼的耗氧率随盐度的升高而下降 ,随温度的升高而上升 ,耗氧率遮光比在自然光下要低。同时还测定了幼鱼的窒息点。     

Androgen secretion activity of recombinant follicle-stimulating hormone of Japanese eel, Anguilla japonica in immature and maturing eel testes

The androgen secretion activities of recombinant Japanese eel follicle-stimulating hormone (rjeFSH) were investigated in immature and maturing eel testes. The rjeFSH stimulated testosterone (T) and 11-ketotestosterone (11-KT) secretion in immature testis but not in maturing testis. This result suggests that eel FSH plays an important role through the sex steroid secretion in immature testis rather than in maturing testis.     

Fluorescence localization of epidermal cathepsins L and B in the Japanese eel

Histochemical localization of proteolytic activities in the dorsal epidermis of Japanese eel was demonstrated by fluorescent microscopy utilizing 4-methoxy-2-naphthylamide (4M$\beta$NA) derivatives as substrates and 5-nitrosalicylaldehyde as a trapping agent. Carbobenzoxy-L-phenylalanyl-L-arginyl-4M$\beta$NA (Cbz-Phe-Arg-4M$\beta$NA) and Cbz-Arg-Arg-4M$\beta$NA were used for direct detection of cathepsins L and B activities, respectively, in fresh frozen sections and unfixed cells of the eel epidermis. The fluorescing areas, where Cbz-Phe-Arg-4M$\beta$NA was hydrolyzed by cathepsin L, were shown in mucus secretory cells and club cells and broadly around skin surface. The fluorescing areas due to Cbz-Arg-Arg-4M$\beta$NA hydrolysis by cathepsin B were localized similarly in these tissues. The fluorescing intensity for both catheptic activities in mucus secretory cells was higher than that in club cells, where small fluorescing granules were distributed. These results indicate that eel cathepsins L and B are stored in epidermal secretory cells at different levels and probably serve as defense factors before or after secretion by these cells. Abbreviations: Cbz – carbobenzoxy; 4M$\beta$NA – 4-methoxy-2-naphthylamide; NSA – 5-nitrosalicylaldehyde.     

Comparison of glass eel stages of American eel and speckled worm eel in a northeast Florida estuary

Since 2001, a glass American eel, Anguilla rostrata (Lesueur), survey has been conducted at Guana River Dam, Florida, USA. Although present in earlier samples, glass speckled worm eel, Myrophis punctatus Lütken, were not identified until a large proportion appeared in samples in 2006. To determine whether the species could be separated in historical samples, weight–length relationships were compared between the two species. Then, with data from the full survey period, relationships between catch rate and annual percent contribution of glass speckled worm eel were explored in relation to temperature, water flow and salinity. Glass speckled worm eel were found to weigh significantly less and had distinguishing physical characteristics that could be used for identification. Glass speckled worm eel constituted between 0 and 34% of the catch, with no detectable pattern in timing or abundance among years. In 2006 and 2012, which had the two largest catches of glass speckled worm eel, relationship between nightly catch rate and measured environmental variables was inconsistent, even when measured variables had a similar range in values. It is recommended that south‐eastern, including Gulf of Mexico, USA, states restrict harvest of glass American eel to prevent the incidental capture and exportation of glass speckled worm eel. Efforts should be made to ensure proper species identification in glass eel surveys.     

外源性促性腺激素诱导日本鳗鲡卵子发生和卵巢发育成熟的机制

用兔抗促黄体素生成素受体(LHR)或称绒毛膜促性腺激素受体(CGR)、雌激素受体(ER)和孕激素受体(PR)的抗体对LHR,ER和PR进行免疫组织化学定位。目的在于揭示外源性促性腺激素(鲤鱼脑垂体提取物,CPE和人绒毛膜促性腺激素,hCG)诱发日本鳗鲡卵子发生和卵母细胞成熟的内分泌机制。结果表明,注射激素前后卵巢发育和卵子发生出现了十分显著的变化。卵巢组织学切片观察显示激素处理前鳗鲡卵巢发育处于卵黄发生早期,卵母细胞平均直径(220±0.01)μm。第一次注射这两种激素后10d,实验组卵母细胞中卵黄核分散在核的周围,核仁数量显著增加,多达18～20个左右,而对照组8～10个。第3和4次激素处理后,卵母细胞发育进入卵黄发生早-中期至中期,卵黄颗粒数量增加。第6和7次激素处理后,卵母细胞进入卵黄发生中后期到成熟期,卵母细胞胞质中充满卵黄颗粒,胞径和核径增加,分别为(570±1.39)μm和(128±1.19)μm,而对照组没有变化。其次,免疫组织化学染色结果显示LHR、ER和PR均定位在鳗鲡卵巢中卵母细胞胞质、核膜、核质、卵被膜和体细胞上。这里值得指出的是,从第3次和第4次激素处理后,这三种受体的定位各有特点,L...     

池塘养殖鳗鲡初孵仔鳗生长发育和卵黄消耗的研究

本文初步研究了池塘养殖日本鳗鲡初孵仔鳗生长与卵黄消耗的关系。在水温(24±1)℃下,仔鳗发育至7~8日龄,口、肠道、肛门、胸鳍等器官形成。仔鳗全长与卵黄的关系为:(1)0~18 d仔鳗全长随日龄的增加而有不同程度的增长;其中0~7 d生长快,平均日增长0.57 mm/d,8~10 d,平均日增长0.13 mm/d,11~18 d,平均日增长0.09mm/d;(2)0~10 d,仔鳗以内源性营养方式吸收营养,仔鳗全长与卵黄消耗呈指数相关,可用公式Y=1.5879e1.4371L(R2=0.8979)表示,仔鳗卵黄消耗与日龄可用公式Y=0.0852e-0.5547L(R2=0.9702)表示;10 d后仔鳗卵黄消失。     

Artificial induction of maturation and fertilization in the Japanese eel, Anguilla japonica

Repeated injections of salmon pituitary extract (20 mg per fish per week) induced vitellogenesis in feminized, cultivated Japanese eels (Anguilla japonica). Oocytes were attained at the migratory nucleus stage after 11 or 12 injections. Addition of 17,20-dihydroxy-4-pregnen-3-one (DHP) into the incubation medium induced germinal vesicle breakdown (GVBD) in the oocytes at the migratory nucleus stage. An injection of DHP (2 µg g^-1 BW), given 24h after an injection of salmon pituitary extract (20 mg fish^-1), succeeded in inducing maturation and ovulation in females which contained occytes at the migratory nucleus stage. Most fish ovulated 15–18h following the DHP injection. Eggs that were ovulated within 15h after the DHP injection showed high fertility and hatchability, but eggs ovulated 18 or 21h after the DHP injection, showed considerably lower fertility and hatchability. A delay between ovulation and stripping of the eggs rapidly decreased both the fertility and hatchability within 6–9h after ovulation, indicating that artificial fertilization must be carried out immediately after ovulation. Repeated injections of human chorionic gonadotropin (hCG) at a concentration of 1 IU g^-1 BW week^-1 induced spermatogenesis, spermiation, and the acquisition of potential for sperm motility in cultivated males. Most males spermiated after the fifth or sixth injection of hCG, and the milt weight gradually increased and remained constant (1–2 g) from the 11th to 31th injection. Sperm motility peaked 24h after each weekly injection, and decreased from the 3^rd day after the injection. Potassium ions are an essential constituent for the maintenance of motility in the eel spermatozoa. Artificial seminal plasma containing 15.2 mM KCl is applicable as a milt diluent. Using these techniques developed for female and male eels, we have succeeded in obtaining many fertilized eggs from cultivated eels.     

Decreasing dietary lipids improves larval survival and growth of Japanese eel Anguilla japonica

Shark eggs-based diet is the only diet by which eel larvae can grow to glass eels in captivity. However, the high level of lipids in the diet is suggested to negatively affect eel larvae. This paper examines the effect of defatted shark eggs (DSE) and hen egg yolk (HY) on growth and survival of larvae of Japanese eel Anguilla japonica. Lyophilized shark egg and commercial HY were defatted with n-hexane, and four experimental diets were prepared using both defatted and untreated shark eggs and HY. Larvae were reared for 3 weeks by feeding the experimental diets. The highest survival rate was observed in the larvae fed DSE, and larvae fed HY showed the lowest survival rate. The best growth was found in larvae fed DSE, followed by shark eggs and defatted HY, and the worst growth was in HY-fed larvae. These results show that decreasing dietary lipids improves the nutritional value of both shark eggs and HY for eel larvae. Regulation of the dietary lipid level may positively affect the larval performance of eels by combination of ingredients with a low lipid content.     

人工条件下日本鳗鲡胚胎及早期仔鱼发育的生物学特征

在多年对日本鳗鲡人工育苗技术研究的基础上,通过采用对胚胎和早期仔鱼的活体观察、计算机图文分析以及组织切片等方法,系统研究了鳗鲡胚胎及孵化后19d前仔鱼的生物学特性。研究结果表明,在水温(23±0.5)℃条件下,日本鳗鲡从受精卵到仔鱼出膜需约38h30min,总积温为885.50℃·h。其特征与一般硬骨鱼类胚胎发育基本相同,鳗鲡胚胎发育可分为14期,为典型的盘状卵裂,原肠作用通过细胞的下包和内卷完成。但日本鳗鲡早期仔鱼与一般硬骨鱼类的仔鱼存在较大差异,表现为无色透明的血细胞和S形心管;开放型的口腔,发达且不能闭合的上下颌和3对颌齿;孵化后3～9d在身体两侧出现由神经细胞和支持细胞构成发达的感觉丘,其数目为6～9对不等;以及形态不断变化的尾垂体等生物学特性。分析认为这些生物学特性决定了日本鳗鲡仔鱼的孵化环境可能相对较稳定,以及仔鱼摄食方式可能为触碰后咬食的方式。     

Immunochemical sexing of living yellowfin tuna, Thunnus albacares (Bonnaterre), using a vitellogeninlike protein

Sexing of living yellowfin tuna, Thunnus albacares (Bonnaterre), was carried out based on immunochemical detection of vitellogenin (VTG), a female-specific protein. Tissue samples were taken from swimming fish in a sea pen by a hand-made apparatus. Recently fertilized eggs were also collected before tissue sample trials. A double immunodiffusion pattern showed that the egg homogenate of T. albacares reacted with rabbit antibody against the purified VTG of greater amberjack, Seriola dumerili (Risso). This suggests that the VTG of T albacares has an antigenicity similar to that of S, dumerili and that this antibody can be used to detect T albacares VTG. Immunodot-blotting analysis revealed that, out of the two fish tested, the tissue sample from one, but not the other, reacted with this antibody, suggesting that the former was a mature female, and the latter a male or immature female. Examination of the fish gonads revealed an ovary and a testis, respectively. Immunological and morphological results corresponded completely. Therefore, the method used in the present study is available for discriminating the sexes of T albacares.     

日本鳗鲡早期幼苗趋光性及视觉发育特征

研究了出膜后第1～9 d的日本鳗鲡(Anguilla japonica)早期幼苗,在不同光强度下趋光性分布及视觉发育组织学特征。结果显示:在8900 lux的强光条件下,日本鳗鲡早期幼苗随着发育,表现出明显的负趋光性。到9 d时,占存活幼苗52.2%的幼苗都集中在最暗的110 lux区域;而在2300 lux的弱光条件下,表现出趋弱光性。到9 d时,占存活幼苗44.6%的幼苗都集中在25 lux区域。视网膜发育的组织学观察发现,日本鳗鲡幼苗的视网膜发育首先出现的是外界膜和内界膜,最后出现的是外丛层。幼苗的整个视网膜结构到9 d时,发育为9亚层结构。视网膜中最发达的是外核层,由3种细胞构成。发育最快的是内核层,由2种细胞构成。结果表明,与日本鳗鲡早期幼苗趋光性有关的因素包括视网膜分层、视网膜厚度、内核层细胞数等多种因素。另外,本研究还探讨了日本鳗鲡幼苗在自然界可能生存的水深和光环境。