僧帽牡蛎三倍体和二倍体的生长比较↑（*）

１９９３ 年采用高温（３７ ℃） 休克处理受精卵１５ ｍｉｎ ,抑制第一极体排放诱导出僧帽牡蛎三倍体,在早期幼虫阶段,处理组幼虫平均壳长显著较对照组大,经过１ 年的养殖,三倍体壳长、壳高、软体部湿重和干重分别比二倍体增加１３ ．２ ％ 、８ ．９ ％ 、１２４ ．６ ％ 和８７ ．１ ％ 。１９９７ 年采用０ ．５ ｍｇ／Ｌ 细胞松弛素Ｂ 在５０ ％ 受精卵出现第一极体时处理受精卵１５ ｍｉｎ 诱导出僧帽牡蛎三倍体,经过１４ 个月养殖,三倍体壳长、壳高、体重和软体部湿重分别比二倍体增加３９ ．２６ ％ 、１７ ．２５ ％ 、７２ ．５０ ％ 和１１８ ．４２ ％ ,除个别月份外,三倍体和二倍体的生长差异显著（ Ｐ＜ ０ ．０５） ,僧帽牡蛎三倍体较二倍体具有明显的生长优势。     

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。     

三倍体长牡蛎(Crassostrea gigas)不育性研究

三倍体长牡蛎(Crassostrea gigas)雌性怀卵量在0～2000万,雄性精量在0～1亿,大大低于正常二倍体。三倍体生殖腺发育受阻,大部分停留在增殖期和休止期,一部分虽发育至生长期和成熟期,但与同期的二倍体比较,生殖腺发育差。部分三倍体配子能完成受精作用,并发育至D形幼虫,但不能继续正常发育。卵子减数分裂后为含14～16条染色体的非整倍体。因此,三倍体长牡蛎是不育的。     

人工诱导长牡蛎三倍体发生技术

长牡蛎 Ｃｒａｓｓｏｓｔｒｅａ ｇｉｇａｓ（Ｔｈｕｎｂｅｒｇ）又名太平洋牡蛎，是目前牡蛎养殖的优良种类之一。二倍体长牡蛎在成熟期为形成生殖腺消耗体内贮存的糖元，且排放精卵后肉质变劣，大大降低了商品价值。三倍体牡蛎生殖腺不发育，到个体成熟时不进行有性生殖，生长速度和干肉重量明显超过二倍体牡蛎。笔者在长牡蛎三倍体育苗与养殖技术方面进行了初步探索，现将其中较成熟的操作流程整理如下。１ 受精卯的制备 采用本地海区养殖的１．５～２龄亲贝，选择体大、强壮、肥满度好的个体，冲洗干净进行解剖。将已解剖的、　亲贝分类…     

团头鲂同源四倍体、倍间三倍体与二倍体红细胞的形态特征比较

通过比较同源四倍体、倍间三倍体与二倍体团头鲂（Megalobrama amblycephala）的红细胞、红细胞核的形态大小差异,探讨不同倍性团头鲂的红细胞形态遗传特征。结果表明：（1）团头鲂多倍体（同源4n和倍间3n）的红细胞数量显著低于二倍体（2n）团头鲂（P〈0．05）。其中,倍间三倍体团头鲂每毫升血液的红细胞数为二倍体的78．95％,而同源四倍体的红细胞数量下降幅度较大,仅为2n团头鲂的56．87％。（2）团头鲂同源四倍体、倍间三倍体的红细胞短径和长径、核短径和长径、红细胞表面积、红细胞体积和核体积均依倍性增加而显著增大（P〈0．05）;其中,又以核体积增大最为显著,同源四倍体、倍间三倍体的核体积分别为2n团头鲂的1．97倍和1．39倍。（3）团头鲂同源四倍体每毫升血液红细胞的总体积和总表面积,均明显低于二倍体和倍间三倍体团头鲂（P〈0．05）;倍间三倍体每毫升血液红细胞的总体积要多于二倍体团头鲂（P〈0．05）,但红细胞的总表面积在倍间三倍体与二倍体团头鲂间不存在显著差异（P〉0．05）。（4）在团头鲂同源四倍体中还观察到一定比例的异常红细胞现象。     

太平洋牡蛎二倍体和三倍体卵母细胞发育的超微结构

本文利用电子显微镜比较研究了太平洋牡蛎二倍体和三倍体卵母细胞发育过程中细胞和各细胞器的超微结构变化。结果表明,在卵黄形成期,二倍体卵母细胞呈椭圆形,细胞器发达,细胞外布满微绒毛,卵黄粒多并均匀分布于细胞质中,卵母细胞生物合成旺盛,代谢活动强;而三倍体卵母细胞呈长条形或不规则形,细胞器少,不发达,细胞外未观察到微绒毛,卵黄粒少,有些细胞的卵黄粒畸形,卵母细胞生物合成及代谢弱。     

不同育性长牡蛎性腺发育过程中糖原和脂质的组织化学研究

为探讨不同育性长牡蛎(Crassostrea gigas)性腺发育过程中主要供能物质的利用和变化差异,利用过碘酸雪夫氏染色(PAS染色)和油红O染色法对不同育性三倍体长牡蛎及二倍体长牡蛎性腺组织进行观察分析。结果显示,增殖期,二倍体和三倍体长牡蛎性腺结缔组织中均含有大量糖原和脂质。随着性腺发育,二倍体和可育型三倍体(3nα型)长牡蛎性腺结缔组织中糖原含量明显下降,到成熟期几乎检测不到糖原,说明性腺结缔组织中的糖原为配子发生提供能量。脂质主要存在于性腺结缔组织和卵母细胞的细胞质中,说明脂质是卵细胞发育的重要组成物质。雌性二倍体和3nα型长牡蛎性腺结缔组织中的脂类含量随着性腺发育并未发生明显下降,推测糖原可能转化为脂质以满足性腺发育的物质需求。脂质在二倍体雄性长牡蛎的性腺发育过程中出现明显的减少,说明脂类在雄性性腺发育中的主要功能可能是供能而不是结构组成。糖原和脂类作为长牡蛎性腺发育过程中重要的供能和组成物质,在不育型三倍体(3nβ型)长牡蛎性腺发育过程中含量没有发生明显的变化,这与3nβ型配子发生受阻密切相关。推测3nβ型长牡蛎由于配子发育受阻,性腺发育初期积累的糖原和脂质并未分配至生殖细胞,而留在结缔组织中,从而使其在繁殖季节仍然能够保持快速生长。研究结果为牡蛎生殖发育调控机制及育性控制育种提供重要信息。     

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。     

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…     

长牡蛎中国群体和美国群体杂交效应与三倍体的优势

将长牡蛎中国群体二倍体分别与美国群体二倍体和四倍体进行杂交,实验共设置4组,分别为杂交二倍体组、杂交三倍体组、中国二倍体组和美国二倍体组,比较了各实验组卵裂率、D幼率、D形幼虫大小及幼虫期、稚贝期的壳高生长、存活率等生物学指标,并估算杂交二倍体的杂种优势率和杂交三倍体的三倍体优势率。结果表明,杂交二倍体幼虫壳高生长的杂种优势率不明显,平均杂种优势率为1.21%,幼虫的存活率及稚贝的壳高生长表现出明显的杂种优势,平均杂种优势率分别为34.47%和20.39%。杂交三倍体的D形幼虫大小、幼虫和稚贝的壳高生长、存活率均表现出三倍体优势,D形幼虫大小三倍体优势率为5.19%,幼虫期壳高生长和存活的平均三倍体优势率分别为4.00%和19.92%,稚贝壳高生长和存活的平均三倍体优势率分别为30.18%和54.43%,200日龄,杂交三倍体鲜体质量的三倍体优势率为202.96%,存活三倍体优势率为73.60%。此外,稚贝期的杂交二倍体生长性状的杂种优势率和杂交三倍体的三倍体优势率均高于幼虫期的优势率。研究表明,中、美两地理群体杂交获得的三倍体长牡蛎子代生长和存活性状都比二倍体优良。杂交三倍体的优良性状主要是三倍体优势,杂交优势的贡献率还有待进一步实验证实。     

The reproductive physiology of three age classes of adult female diploid and triploid brook trout (Salvelinus fontinalis)

Triploid female fish show impaired gametogenesis and are unable to produce viable offspring. The reproductive physiology of artificially-induced triploid female salmonids has been well described up until the time of first sexual maturation in diploids, but few reports exist for older triploids. This study reports the influence of triploidy on growth, ovarian development and reproductive endocrinology among three age classes of female brook trout (Salvelinus fontinalis) in comparison to sibling diploids. Triploids were larger than diploids for most of the study period, but the difference was statistically significant only during maturation and spawning of 2⁺ diploids. Plasma estradiol-17 (E₂), testosterone (T) and vitellogenin (VTG) levels in triploids were generally lower than in diploids, and VTG was the only parameter to show seasonal fluctuations resembling those of diploids. Triploids showed significantly lower GSI and total oocyte number than diploids of similar age, and only half of all triploids sacrificed during the study (n=56) had developing oocytes in their ovaries. At age 3⁺, 13 of 19 triploid females had oocytes at various stages of development, including perinucleolar, yolk vesicle and yolk globule stages. In addition, three of these fish had collectively produced 72 mature stage oocytes. Thus, whereas diploid brook trout can produce mature oocytes as two-year-olds, triploids cannot do so until four years of age, with the number of mature oocytes being greatly reduced.     

Triploid hard clams Mercenaria mercenaria produced by inhibiting polar body I or polar body II

Two types of triploid hard clams Mercenaria mercenaria were produced by inhibiting polar body I (PB1) or polar body II (PB2) with cytochalasin B. Treatments were applied at 22–23°C, with PB1 inhibition starting at 4–7 min postfertilization and ending when PB2 was first observed in control groups, and with PB2 inhibition starting at 17–23 min postfertilization and ending when 80% of control eggs released PB2. Triploid induction success was evaluated by chromosome counting in 2–4 cell embryos and by flow cytometry at larval and juvenile stages. PB2 inhibition produced more triploids (82%–100%) than PB1 inhibition (71%–83%), although the difference was not significant (p ≥ .088). Triploid percentages in PB1‐ or PB2‐inhibited groups showed a small but insignificant decline during the first 6 months. At month 3, PB1 and PB2 triploids were not different from their within‐group diploids, but significantly larger than control diploids; PB1 triploids were significantly larger than PB2 triploids (p ≤ .003). At month 6, PB1 triploids were not different from either within‐group or control‐group diploids, while PB2 triploids were significant larger than both within‐group and control diploid; PB1 triploids were smaller than PB2 triploids. At month 16, PB1 and PB2 triploids in one remaining replicate were not different from their within‐group diploids. Overall, this study shows that triploids can be efficiently produced by PB1 or PB2 inhibition, and their growth performance relative to diploids is variable depending on age and replicates or parental genotype.     

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.     

Larval metamorphosis and morphological characteristic analysis of triploid shrimp Fenneropenaeus chinensis (Osbeck, 1765)

Both MI and MII triploids were successfully produced by heat shock in Chinese shrimp Fenneropenaeus chinensis. The inducing conditions for MI and MII triploids were optimized. The highest inducing rate obtained for MI triploids reached more than 90%, and that for MII triploids reached nearly 100% at the nauplius stage as evaluated using flow cytometry. Comparisons of survival rates at larval stages between triploids and diploids or diploids experiencing treatment and diploids without treatment were performed. At larval stage from nauplii to postlarvae, heat shocks lowered survival at larval stages even if the ploidy was not changed. Ploidy did not affect shrimp larvae survival, and no significant difference was found in the survival of shrimp larvae between MI and MII triploids. Highly significant differences were observed in the morphology of triploids and diploids, and no apparent difference was found in the morphology of MI and MII triploids at the grow‐out stages. Discriminating formulae for triploid and diploid shrimp at grow‐out stage were developed and could be used to distinguish triploids from diploids based on morphological parameters. MI and MII triploids of shrimp have the potential to be used in aquaculture.     

The reproductive cycle of the ‘blunt’ sea urchin

The reproductive cycle of the blunt sea urchin, Sphaerechinus granularis (Lamarck), was studied in the Bay of Brest, West Brittany, from monthly samples by examining seasonal changes in gonadal index, in the stages of gonad development and in oocyte diameters. This echinoid exhibited an annual cycle of gonadal growth, with a short breeding season occurring in May and June, and a decrease in gonadal index during the winter attributed to the use of nutrient resources. Histological examinations defined long mature stages during autumn, winter and early spring, where the vitellogenic oocytes accumulated in the lumen of the gonad. Changes in oocyte diameters showed a short pre-spawning period in April and May. The brief breeding season was followed by rapid post-spawning recovery and gonad growing stages during the summer, where the gonads mainly grew through the development of nutritive tissue. The features of this reproductive cycle show that the species can be commercially exploited and these results are used to make recommendations in terms of the management of S. granularis fisheries in Brittany. The influence of temperature on the initiation of spawning was noted again. This exogenous factor could be monitored to improve gonad production and availability of mature gametes in mariculture projects. © Rapid Science Ltd. 1998     

Comparative Growth of Diploid and Triploid Asian Catfish Clarias macrocephalus in Thailand

Wild caught Asian catfish were spawned manually following HCG injection, and a portion of the eggs were subjected to cold-shock at 4 C for 15 min within 2-min post-fertilization. Nuclear diameter measurements of cold-shocked fish revealed that 96% were triploids (3N), while non-shocked fish were all diploids (2N). During larval and fry culture (first 26 d), triploid fish mortality was =50%, while diploid mortality was =25%. Following 8-mo culture in tanks at three stocking densities, triploid fish survival was significantly greater ( P < 0.05), than diploids, with 84.0% and 57.3%, respectively. Triploid live weight was also significantly greater than diploids, with 69.2 and 45.9 g averages, respectively. Ninety-two percent of diploids had welldeveloped gonads after 8 mo; whereas none of the triploids had mature gonads. Gonads were undifferentiated with 31% of the triploids. These sexually undifferentiated fish had greater growth rates than male or female triploids, and greater growth than all diploids. Carcass weight (gutted) of triploids was 95.8% of live weight, compared with 92.5% for diploids. Lastly, triploids had very few deformities compared with diploids, with 1.3% and 17.6%, respectively. Deformities included curved spines, and humped backs just posterior of the head.     

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。     

Growth and gonadal development in diploid and triploid turbot (Scophthalmus maximus)

This study determined the effect of triploidy on the survival, growth and gonadal development of turbot from 6 to 48 months of age. From 6 to 24 months of age (first sexual maturity), survival was similar in both ploidies (P > 0.05). From 24 to 48 months of age, after the first sexual maturity, survival was 91.9% in diploids and 100% in triploids, which did not exhibit the post-spawning-associated mortality. Growth was similar for both ploidies during the first year of life. After that, triploids grew significantly (P < 0.05) more that diploids, with more marked differences after each spawning season. From 24 to 48 months, the average weight difference between both ploidies was 11.4 ± 1.9%, ranging from 4.3 to 23.0%. At 47 months of age, the biomass of triploids was 10.3% greater in total weight and 14.3% greater in eviscerated weight. Gonads of triploid males were similar to that of diploids, whereas in triploid females, they were significantly smaller and rudimentary. A histological analysis carried out at 47 months of age showed complete sterility of triploids in both sexes. Sex ratio was 1 male (M):0.6 female (F), for diploids, significantly (P < 0.05) different from 1:1, and 1 M:3.3 F for triploids, significantly (P < 0.05) different from 1:1 and from the diploids. Since females grow more than males, culture of triploids benefited from the high female ratio, which helped to reduce size dispersion. In addition, their sterility allowed better performance by avoiding the reduction in growth that takes place during the spawning periods. Together, these observations indicate that triploidy induction can be an interesting option for turbot aquaculture, especially for the production of large-size fish of more than 2 years of age.     

Added value from an added chromosome: Potential of producing large fillet fish from autumn to spring with triploid rainbow trout,Oncorhynchus mykiss

Three‐summers‐old all‐female triploid and diploid rainbow trout were compared after one on‐growing season in sea net cages. Slaughter traits of round weight, gutted weight, fillet weight, carcass% and fillet% were measured at three times in November 2017, January and April 2018. The triploid group had lower daily growth coefficient mean (4.25) and higher feed conversion ratio (1.18) than diploids (4.48 and 1.05, respectively) during on‐growing (June–November). In November, no difference of means was found between mature or immature diploids and triploids for any of the weight traits when the effect of vertebrae defects was statistically removed. However, the triploids had attained higher means than mature or immature diploids in gutted and fillet weight by January, suggesting that the loss of muscle mass during early winter was lower in triploids. Sexually maturing diploids (46%) had lower slaughter yield means compared to triploids or immature diploids at each measurement time, and these differences also increased during overwintering. Instead, the means of yield traits remained similar between the triploid and immature diploid groups through the winter. Likewise, fillet redness remained at equally high level in triploids and immature diploids, whereas in maturing diploids this attribute decreased substantially during overwintering. The triploid group had a higher incidence of vertebral defects (12.0%) than diploids (5.3%). The present results demonstrate the potential of triploid trout in producing large‐sized (>2 kg) fillet fish until spring markets. However, more detailed investigations are needed, particularly regarding the animal health and growth efficiency in triploids, relative to their diploid counterparts.     

Growth and gonadal development in diploid and triploid Atlantic cod (Gadus morhua)

Induction of triploidy has been suggested as an effective tool to prevent spawning of farmed fish. This experiment examined the growth potential of triploid cod when reared communally with diploid ones after the juvenile stage. Pressure treatment was used to induce triploidy in a batch of cod eggs in April 2009. The resulting offspring were reared separately from their diploid counterparts until they reached the proper size for PIT tagging. At the age of 8 months, an equal number of 115 diploids (135.5 ± 3.95 g) and triploids (93.6 ± 2.63 g) were communally reared in a circular flow-through tank until the age of 22 months. By the end of this rearing period, diploids (1,002.4 ± 39.9 g) were significantly heavier than triploids (654.6 ± 27.7 g), but the specific growth rate did not differ significantly during the growth trial. Gonadal development at the age of 22 months was also lower among triploids than diploids, especially for females (5.3 and 91.9 %) but also for males (32.5 and 72.7 %). Sterility among female triploids was evident by the reduced size and dysfunctional gonads, but gonadal development in male triploids was less suppressed. Prevalence of body deformities was, however, significantly higher among triploids (62.6 %) than diploids (33.9 %). Higher prevalence of deformities in triploid cod underlines the need for further fine-tuning of the triploidization procedure or finding other methods of sterilization. At present, triploid cod are still far from being established as an alternative for commercial production.