科威特培育出三倍体罗非鱼

罗非鱼性成熟早,导致在养殖池中大量繁殖,造成群体拥挤而阻碍生长。由于雄鱼比雌鱼有更大的生长潜力,许多人致力于研究生产全雄鱼群体,用于养殖。但是,消费者对食用鱼使用雄性激素处理表示担心。科威特水产研究人员提出了用染色体操作技术来诱导全雌不育三倍体的方法,因为不育的三倍体显示了较快的生长速度。生长试验表明,雌性三倍体比性反转的单性雄鱼生长快14％,比雄性三倍体生长快23％。诱导全雌不育三倍体的程序如下:正常的二倍体雌鱼     

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。     

牙鲆三倍体批量化诱导及其生长和性腺发育观察

对牙鲆三倍体鱼苗的诱导、生长及性腺发育进行了一系列研究,发现在14.8～15.5 ℃的海水温度条件下授精,授精后3 min将受精卵放入3 ℃的海水中冷休克处理45 min,可以得到牙鲆三倍体;120日龄、348日龄、630日龄时对三倍体和二倍体的全长和体质量进行测量,发现在120日龄时二倍体与三倍体体质量差异不显著,348日龄时三倍体体质量明显大于二倍体体质量(P<0.05),630日龄时三倍体的全长和体质量均高于二倍体,且差异极显著(P<0.01)。通过比较630日龄时的性腺指数,发现二倍体牙鲆雌性和雄性的性腺指数分别是三倍体的3.3倍和3.1倍,三倍体牙鲆性腺明显小于二倍体,二倍体和三倍体卵巢指数之间及精巢指数之间差异均极显著(P<0.001);组织切片观察显示,在二倍体卵巢中可见正常卵母细胞,三倍体卵巢处于未分化的卵原细胞阶段,三倍体精巢中有精母细胞,但数量明显少于二倍体精巢。结果表明,研究采用的方法能够大量获得诱导率为100%的三倍体牙鲆鱼苗,三倍体成鱼性腺发育不良,并且其生长速度显著快于二倍体对照,适合在生产中推广。     

半滑舌鳎三倍体鱼苗的人工诱导与鉴定

针对半滑舌鳎雌雄个体生长差异过大、雌性性腺发育成熟后腹部凸起影响舌鳎生长和商品鱼质量等问题,开展了人工诱导半滑舌鳎三倍体的研究。采用静水压处理抑制半滑舌鳎受精卵第二极体排出进行染色体加倍,筛选出有效的静水压处理强度及其持续时间。结果表明,孵化水温23 ℃左右时,授精后5 min,采用36 MPa的静水压压力,休克处理4 min,三倍体诱导率最高,达到100%。采用该诱导条件,大批量获得了半滑舌鳎三倍体鱼苗。采用流式细胞仪分析了三倍体鱼苗细胞DNA含量,表明三倍体鱼苗细胞DNA含量为二倍体对照鱼苗的1.5倍。通过染色体分析表明,三倍体鱼苗的染色体数为63条,而二倍体鱼苗的染色体数为42条。     

二、三倍体鲤性腺发育的比较研究

二、三倍体鲤鱼在单养时，三倍体鱼的性腺重和GSI均低于二倍体，经t检验，性腺重数值差异显著（P＜0．05）、GSI数值差异极显著（P＜0．001）；在混养时，三倍体鱼的性腺重和GSI均低于二倍体，差异均显著（P＜0．05）。雄性三倍体鲤鱼的精巢发育比较一致，均完成了精子发生过程     

渔业生物学

021134自转基因三倍体泥鳅生长快、个大且不育”Aeeelerated growth,gigantism and1 ikely sterility in autotran笔enie triploid mudloach斌sg“r1l“mizolePis〔刊,英〕用amYK,Cho HJ,Cho YS.二// J .WorldAquae.S叉.一2001,32(4)一353一363 转基因鱼因其生长快而缩短了养殖时间,具有明显的经济效益。对韩国重要经济鱼类泥鳅进行了三倍体诱导试验,细胞DNA、染色体等均被证实成功地诱导了三倍体。南方杂种泥鳅和自转基因三倍体泥鳅的GHm RNA与自转基因二倍体泥鳅的表现型近似。自转基因三倍体泥鳅的生长速度是普通二倍体泥鳅的22…     

养殖条件下鲻鱼生长发育的初步研究

鲻鱼（Mugil cophalus Linnaeus）是鲻科鱼类中分布最广且最早被人类利用及研究的对象之一，其个体大、生长快、抗病性强、食物链级次低，可在淡水、半咸水及海水中正常生长。有关鲻鱼生长发育的研究，国内外学者已做了不少工作。鲻鱼的生长情况与鲻鱼养殖的经济效益有着直接的关系，同时也影响到鲻鱼性腺的发育，关系到鲻鱼人工繁殖工作的研究。     

虹鳟鱼三倍体的鉴定

三倍体是非偶数染色体组,阻碍了生殖细胞正常的减数分裂,结果常常导致性腺不能发育,不育的三倍体经济鱼类因为没有性成熟阶段性腺发育的能量损耗,所以有以下优点：避免了性腺发育阶段和产卵季节鱼肉质量下降并延长了上市时间；避免了性腺发育阶段的生长停滞和死亡率增加；缩短了养殖周期,减少了养殖成本并可养成大型个体。用人工的方法获得三倍体可通过两条途径,一条是直接途径,即用物理或化学的方法对受精卵进行处理,阻碍第二极体外排；另一条是间接途径,即首先诱导获得四倍体,待四倍体个体性成熟后再与二倍体个体自然交配,获得三倍体。但目前人工诱导的三倍体绝大部分是采用第一条途径获得的,这是因为阻止第二极体外排的方法比较成熟,尤其是虹鳟。     

养殖鱼类的三倍体育种技术与应用综述

多倍体鱼类在生长、群体产量及抗病力等各方面都具有二倍体鱼所无法比拟的优势，有很好的潜在利用价值。三倍体鱼指的是含有三套染色体组的多倍体鱼，具有生长速度快、肉质好、抗逆性强、不育不干扰鱼类资源等优点。三倍体技术成为养殖鱼类育种的重要手段，近年来，与其相关的研究很多，本文就其原理和应用作了回顾。     

大西洋鲑性腺分化及热休克的影响

大西洋鲑性腺分化发生在孵出45－63天，约495－693度日。经热休克处理获得三倍体稚鱼，性腺在分化之前明显大于正常二倍体。雌性三倍体稚鱼性腺在细胞学分化时受抑，初级卵母细胞不能发育；雄性三倍体稚鱼性腺在细胞学分化阶段生长明显减慢，初级精母细胞尚能发育。     

栉孔扇贝三倍体与二倍体的排泄研究

从温度变化、个体大小和昼夜变化等３个角度研究栉孔扇贝（Ｓｃｈｌａｍｙｓ ｆａｒｒｅｉ）三倍体与二倍体的排泄率。结果表明，栉孔扇贝三倍体与二倍体的排泄率均随温度升高而增大；不同壳宽排泄率（Ｎ）与温度（ｔ）的关系为：Ｎ＝７．２０３３ｅ＾０．１２１５ｔ，Ｒ＾２＝０．８４４１（三倍体ｓ组，壳宽约３．２ｃｍ）；Ｎ＝８．８１５３ｅ＾０．１０３１ｔ，Ｒ＾２＝０．９８６３（三倍体ｂ组，壳宽约４．３ｃｍ）；Ｎ＝６．     

Ethidium Bromide Nuclear Staining and Fluorescence Microscopy: An Alternative Method for Triploidy Detection in Fish

Abstract.– Erythrocyte nuclei from normal diploid (2N) and cold-shocked triploid (3N) Asian catfish Clarias macrocephalus were comparatively stained with fluorescent stain, ethidium bromide and with a bright-field stain, Giemsa. Stains were applied to dried blood smears on glass slides, with blood taken from fingerling fish without sacrificing the fish. Both stains produced comparable quality visual comparisons, with significant differences ( P < 0.001) between nuclear dimensions of 2N and 3N cells. Triploid nuclei were larger and more elongate than diploid nuclei. Karyotypic comparisons revealed that diploid fish had 54 chromosomes, while triploid fish had 81. Ethidium bromide nuclear staining required only 3 min after slides were air-dried and fixed, while Giemsa staining required more than 50 min.     

Survival, growth and gonad development of triploid red sea bream, Pagrus major (Temminck et Schlegel): use of allozyme markers for ploidy and family identification

Abstract. Triploidy was induced in three full-sib families of sea bream, Pagrus major (Temminck & Schlegel), by subjecting the fertilized eggs to cold-shock treatment at 0°C for a 12 min duration, starting 3 min after insemination. Results showed that this treatment gave 100% triploid fish with reasonable survival rates of more than 61% up to first feeding, Triploidy induction was proved by the use of allozyme markers at three ( ADH *, EST * and 6PGD *) loci and red blood cell size analysis.
At 2 months of age, the survival of the triploid group was lower than that of the diploids but it was comparable at 10 months. No differences in growth rate between diploid and triploid full-sibs were observed up to the age of 10 months. However, differences in body weight between families were noticed both in diploid and triploid groups when cultured in the same net cage. This possibly suggests a high heritability for the trait examined in the broodstock. The gonads of the triploid fish were significantly smaller than those of the diploids, Histological analysis of the gonads revealed sterility in triploid fish.     

二、三倍体乌克兰鳞鲤染色体核型分析

以二、三倍体乌克兰鳞鲤为材料,采用植物血球凝集素(PHA)体内注射,肾组织细胞短期培养,常规空气干燥法制备染色体.其核型分析结果:二倍体乌克兰鳞鲤染色体2n=100核型公式为:26m+30sm+30st+14t,染色体臂数NF=156;三倍体乌克兰鳞鲤染色体的核型公式为3n=150=39m+45sm+45st+21t,染色体臂数NF=234,二、三倍体染色体数之比为1:1.5.二倍体核型与已报道的鲤鱼染色体核型相似.未发现性染色体.     

Induction of triploidy in mud loach (Misgurnus mizolepis) and its effect on gonad development and growth

Triploidy was induced in mud loach (Misgurnus mizolepis) by cold shocking fertilized eggs 5 min post-fertilization at 2°C for 15 to 60 min. Best results were obtained when eggs were shocked for 60 min; 98% of fish examined in that treatment were triploids. Triploidy was confirmed by erythrocyte measurements and chromosome counts. Diploids had 48 chromosomes, while triploids had 72. Histological analysis of 9-month-old triploid ovaries showed an appreciable number of oocytes at the chromatin nucleolus stage with considerable interstitial tissue. However, diploids had well developed oocytes. Diploid testes from diploid males exhibited normal spermatids and spermatozoa, while a few were seen in triploid males. Growth rate was evaluated over a 9-month growth trial. Although male and female triploids were slightly heavier than their diploid counterparts from the third to the ninth month, their growth rates were not significantly different compared to their diploid controls.     

Growth and feeding in juvenile triploid and diploid blacklip abalone, Haliotis rubra (Leach, 1814), at two temperatures

Growth and feeding of juvenile triploid and diploid blacklip abalone Haliotis rubra (Leach, 1814) were investigated at two temperatures of 17 and 21 °C over a 50‐day period. There were no differences in growth between triploid and diploid abalone as measured by shell length and body weight. Both triploid and diploid abalone increased in length but not in weight at 21 °C. Condition indices were similar for triploid abalone maintained at both temperatures; however, those for diploid abalone were significantly higher at 17 °C than at 21 °C. Food intake was significantly greater yet feed conversion efficiency was significantly lower in triploid than in diploid abalone. Both the feeding variables were independent of temperature. On average, diploid abalone were able to convert 1 g of dry food ingested to 0.58 g of body weight, but triploid abalone only 0.44 g.     

Triploidization in rohu × mrigal hybrid and comparison of growth performance of triploid hybrid

The present study was aimed at the identification of treatment optima to induce triploidy in ‘Labeo rohita (rohu) × Cirrhinus cirrhosus (mrigal)’ hybrid using heat shock treatment. The eggs were exposed at four different temperature regimes viz., 38, 39, 40 and 41°C for 1–3 min, applied 3–5 min after fertilization. After 4 min of fertilization, heat shock treatments for 1 and 1.5 min durations were found the best inducing triploidy up to 100% and 96% respectively. Survival rates upto yolk sac absorption were found to be 73% and 71% in rohu and mrigal, 68% and 67% in the reciprocal diploid hybrids and 61% and 60% in the reciprocal triploid hybrids (RTH). Triploidy was confirmed by chromosome counting that revealed the diploid chromosome number of rohu and mrigal at 2n = 50 and in their triploid hybrid chromosome number was found to be 3n = 75. Growth rate of the RTH showed a significant difference (P < 0.05) from the single species and the diploid hybrids. Triploids also showed higher survival rate over the diploids.     

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)。     

Growth and reproductive performance of triploid and diploid blacklip abalone, Haliotis rubra (Leach, 1814)

Growth and reproduction of triploid and diploid blacklip abalone Haliotis rubra (Leach, 1814) were compared in a 30-month study. Triploidy was induced by inhibition of the second polar body formation using 6-dimethylaminopurine (6-DMAP) or cytochalasin B (CB). There were no significant differences in growth and survivorship between triploid and diploid abalone. However, triploid abalone had a more elongated shell and greater foot muscles than diploid abalone. A slightly curvilinear growth in shell length was conformed to all treatments. While diploid abalone had reached sexual maturity and spawned during the study, gonadal development and gamete maturation were abnormal in triploids. Female triploids lacked an apparent gonad at the macroscopic level but microscopic examination revealed that they had a thin layer of oogonia development. In contrast, male triploids were able to form similar-sized gonads to diploids during most of the reproductive period, but with brown-yellow discolouration and stalled gametogenesis at spermatocyte formation. Sex ratio of triploid abalone did not deviate from 1:1. With the onset of sexual maturation, growth and gonadal maturation occurred concurrently in diploid abalone, and there was no indication that growth of (diploid) abalone was reduced.     

病原感染条件下中国对虾二倍体和三倍体血液学变化

以热休克诱导获得中国对虾三倍体，通过口服或注射的方式对二倍体和三倍体中国对虾进行了白斑综合征病毒(WSSV)和鳗弧菌感染实验，研究了病原感染前后二倍体和三倍体中国对虾血液学的变化并首次报道了三倍体中国对虾血细胞的组成。实验结果表明：三倍体和二倍体中国对虾均对WSSV敏感，三倍体未显示出明显的抗性。WSSV感染4d后，对虾外周循环系统的血细胞数量明显减少，三倍体和二倍体对虾血细胞数量均降为对照组的10％左右。对虾血细胞组成发生变化，大颗粒细胞明显减少，小颗粒细胞相应增加，而透明细胞所占比例基本不变；大颗粒细胞和小颗粒细胞内颗粒物质均有所减少，出现空泡或细胞外突起等异常结构。注射鳗弧菌感染后两种对虾的血细胞数量均有所减少，血细胞数量的减少与弧菌的注射量呈正相关关系：三倍体中国对虾血细胞组成与二倍体相似，但其密度仅为二倍体的40％左右：本研究结果为三倍体对虾抗逆特性的研究提供了一定依据和参考。