Verification of ploidy level in sturgeon larvae

Chromosome preparations and assay of the microsatellite locus Afu‐68 were used to determine ploidy in Siberian sturgeon (Acipenser baeri Brandt) and F₁ hybrids of Siberian sturgeon and Russian sturgeon (Acipenser gueldenstaedti Brandt). The chromosome number and microsatellite locus Afu‐68 were compared and these analyses were used for identification of ‘haploid’, ‘diploid’ and ‘triploid’ progeny of the studied cross of A. baeri× (A. baeri×A. gueldenstaedti).     

Growth performance, morphometric traits and gonad development of induced reciprocal diploid and triploid hybrids between the mud loach (Misgurnus mizolepis Günther) and cyprinid loach (Misgurnus anguillicaudatus Cantor)

Two species of loach, the mud loach (Misgurnus mizolepis Günther) and the cyprinid loach (M. anguillicaudatus Cantor), are commercially important in Korea both for food and ceremonial purposes. The mud loach has superior potential for aquaculture in terms of growth, whereas the cyprinid loach has a more desirable body shape and colour. This study was conducted to produce reciprocal diploid and triploid hybrids and to evaluate their potential benefits in culture by examining growth performance, morphometrics and gonad development. Reciprocal diploid and triploid hybridization was performed by artificial insemination without or with the induction of triploidy. The successful formation of diploid or triploid karyogamy was verified by flow cytometric analysis. Body weights of induced reciprocal diploid hybrids were intermediate between those of the two parental species, i.e. hybrids were heavier than cyprinid loach but lighter than mud loach. In contrast, the growth performance of triploid hybrids was similar to that of their corresponding maternal parents. Diploid hybrids showed intermediate morphometric traits between the two species; however, the body proportions of triploid hybrids were more similar to those of the maternal species than the paternal species. Histological analyses indicated that reciprocal diploid hybrids of both sexes were able to reach maturity, as evidenced by the presence of mature oocytes or spermatozoa in the gonad tissues. However, triploid hybrids showed stringent sterility at the gonadic level; the sizes of ovaries and testes were much smaller, and gonad development was abnormal and significantly retarded.     

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．     

5种壳色长牡蛎不同组织常规营养成分分析与评价

为探究DNA甲基化在长牡蛎性腺发育中的表观遗传调控机制,对长牡蛎Htatip2的同源性、系统进化、组织表达以及三倍体性腺可育型和不育型不同发育时期的基因表达和DNA甲基化谱进行了研究。结果显示,长牡蛎Htatip2的保守结构域与美洲牡蛎的Htatip2-like的保守结构域同源性最高。qPCR分析显示, Htatip2在各个组织中均有表达,其中在雌性性腺中的表达量最高。此外,该基因的表达量在可育型三倍体牡蛎性腺中随着性腺发育成熟而升高,在不育型三倍体牡蛎性腺中表达量变化不显著。BS-PCR分析显示,该基因的甲基化水平随着性腺发育成熟而降低,与基因表达水平成负相关性。双荧光素酶报告结果显示,甲基化的Htatip2启动子片段与未甲基化的片段相比,显著抑制了荧光素酶的活性。研究表明,长牡蛎Htatip2的DNA甲基化可能通过抑制基因表达参与了性腺成熟调控。本研究为表观遗传调控机制参与牡蛎性腺发育提供了重要参考依据。     

文蛤二倍体和三倍体染色体核型分析

以担轮幼虫为材料,采用热滴片法制作染色体标本,对二倍体和三倍体的文蛤(Meretrix meretrix Linnaeus)染色体数目及核型进行探讨,结果显示,文蛤二倍体的核型为2N=38,24M 14SM,NF=76;三倍体的核型为3N=57,36M 21SM,NF=114。二者染色体类型基本相同,均未发现有异型和具随体的染色体。另外本研究还分析了文蛤与帘蛤目(Veneroida)及帘蛤科(Veneridae)的其他种类之间的核型异同,为探讨它们之间的亲源关系提供科学依据。     

二倍体和三倍体皱纹盘鲍精子发生过程的超微结构

比较了二倍体和三倍体皱纹盘鲍精子发生过程中细胞和细胞器的超微结构变化。结果表明,二倍体皱纹盘鲍的精子发生经历了精原细胞、初级精母细胞、次级精母细胞、精子细胞和精子5个阶段。其形态结构发生了一系列变化,主要包括：核染色质浓缩、线粒体的发达与融合、顶体形成和胞质的减少。三倍体皱纹盘鲍各种生精细胞的直径和核径均大于二倍体,精原细胞结构与二倍体相似;初级、次级精母细胞的胞质中,除溶酶体外,线粒体、内质网等细胞器少于二倍体,线粒体大小与二倍体没有差别,但形态不典型,层状嵴不发达;三倍体皱纹盘鲍的精子发生停滞在精子细胞阶段,表现出各种畸形状态,很多趋于解体,没有发现成熟精子。     

The physiology of triploid fish: current knowledge and comparisons with diploid fish

This paper is aimed at underlining the limited knowledge available about the physiology of triploid fishes compared with diploids. Whereas many aspects (induction, detection, growth, resistance to diseases, etc.) in the production and rearing of triploid fishes have widely been developed and described in the literature, other numerous questions of ecophysiology remain in abeyance, and the study of triploid cells physiology is still in its infancy. Triploid fishes can be considered as models worthwhile for physiological investigations not only for the economical stake in relation to the development of triploid fishes rearing, but also for the cytological and molecular features of their tissues and organs. The functional implications of these features have been poorly studied although they are potential areas of applied and/or fundamental studies.     

虾夷扇贝二倍体与三倍体脂肪酸组成的比较

采用毛细管色谱柱对虾夷扇贝二倍体与三倍体的脂肪酸组成进行了检测。结果表明 :①两者闭壳肌和外套膜的脂肪酸组成相似 ,特征为多烯酸 (PUFA) >饱和脂肪酸 (SFA) >单烯酸 (MUFA) ,PUFA以n 3系列为主 ,主要脂肪酸为 2 2∶6n 3(2 3.9%～ 2 8.9% )、2 0∶5n 3(17.6 %～ 2 0 .6 % )、16∶0 (11.0 %～ 19.4 % )、18∶0 (7.3%～8.1% )、2 0∶4n 6 (2 .8%～ 5 .5 % ) ;②生殖腺的脂肪酸种类相似 ,但两者多数脂肪酸的含量差异显著 ,其中三倍体的SFA和MUFA含量显著高于二倍体 (P <0 .0 1) ,而PUFA含量显著低于二倍体 (P <0 .0 1)。     

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。     

Test of Mendelian segregation among 10 microsatellite loci in the fourth generation of bester (Huso huso L. ×Acipenser ruthenus L.)

Ten microsatellite loci were tested on Mendelian segregation in the bester – hybrid of beluga, Huso huso L. and sterlet, Acipenser ruthenus L. in the fourth generation. All studied loci showed disomic inheritance and Mendelian segregation of their alleles. The molecular approach by microsatellite DNA analysis represents a reliable tool for interspecific characterization of sturgeons and their hybrids (different alleles of several loci are present in sturgeon fishes, and interspecific hybrids posesses both maternal and paternal specific alleles). This is the first report of the Mendelian segregation testing in interspecific hybrid of acipenserid fishes using microsatellite DNA markers.     

Inheritance of microsatellite loci in polyploid Siberian sturgeon (Acipenser baeri Brandt) based on uniparental haploids

Haploid gynogenesis was induced in Siberian sturgeon (Acipenser baeri Brandt) using UV‐irradiated bester (hybrid of Huso huso L. and Acipenser ruthenus L.) sperm. The inactivated heterologous sperm was used successfully to produce gynogenetic haploids of Siberian sturgeon. All haploid progeny were screened using microsatellite DNA analysis and uniparental transmission in gynogenetic haploids was confirmed. The objective of this study was to obtain haploids of Siberian sturgeon and describe the inheritance of microsatellite loci in haploid embryos of this species. This initial work, based on inheritance in haploid Siberian sturgeon, suggests that three studied microsatellite loci segregate disomically, three tetrasomically and one octasomically.     

Reproductive ability of triploid ornamental (koi) carp (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids and characteristics of their offspring

The purpose of this study was to investigate reproductive ability of backcross triploid koi (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids. These triploids have been obtained by crossing of F₁ hybrid females producing diploid eggs with males of parental species. Triploid hybrid females, when crossed with goldfish or koi males, produced mostly aneuploid fish with ploidy range from approximately 2.2n–3.2n with a mean value 2.5n; some fish in crosses of triploid females with koi males were tetraploid (4.0n). Since analysed fish had in their genomes one haploid set from parental males, the data indicate that triploid hybrid females mostly produced aneuploid eggs with ploidy range from approximately 1.2n–2.2n and a mean ploidy around 1.5n while some eggs were triploid. Triploid hybrid males were completely sterile and have not released any sperm after hormonal injection. Despite their low viability, some aneuploid fish obtained from triploid hybrid females were raised in indoor recirculating systems until the age of 2 years and their reproductive ability has been evaluated. One aneuploid female with ploidy 2.1n produced larvae with ploidy range from 2.9n to 3.4n with a mean ploidy of 3.1n when crossed with a koi male; about 60% of obtained larvae had ploidy from 3.0n to 3.2n. These data indicate that this female produced mostly eggs with unreduced ploidy level.     

The hematology of diploid and triploid transgenic Atlantic salmon (Salmo salar)

This study examined the erythrocyte height/length, hematocrit, total blood hemoglobin concentration and mean cellular hemoglobin content of diploid and triploid transgenic and non-transgenic Atlantic salmon, to clarify the interaction between metabolic rate and hematology in these fish. Triploid erythrocytes were significantly longer and proportionately thinner than diploid erythrocytes for both genotypes. These morphological differences gave triploid erythrocytes an elliptical appearance with the width being ∼53% of the length, whereas diploid erythrocytes tended to be more rounded with the width being ∼62% of the length. As well, diploid and triploid transgenic erythrocytes were significantly shorter and thinner (P<0.0001) than their non-transgenic counterparts. Although not significant, observations using a Channelized Coulter Counter showed that transgenic erythrocytes tended to be higher in count and smaller in volume than non-transgenic erythrocytes of the same ploidy. Transgenics likely produce erythrocytes with higher surface area to volume ratio in response to their elevated metabolic rates. No other major hematological differences were observed between transgenics and non-transgenics of the same ploidy. This revised version was published online in August 2006 with corrections to the Cover Date.     

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。     

Comparison of freshwater and marine performances of all‐female diploid and triploid Atlantic salmon (Salmo salar L.)

The performance of all‐female diploid (AF2N) and triploid (AF3N) Atlantic salmon were compared in fresh water, under commercial production conditions in 1995 and 1996 year classes. The performance of the 1996 year class was also assessed for 14 months in a commercial sea farm. Freshwater mortality was higher in the triploid groups. The majority of losses occurred in the early stages of egg development and during the first feeding period, when the incidence of non‐feeding fry was consistently higher. In growth studies, although diploid fry were significantly heavier during first feeding there were no significant differences in weight between groups some 8 months after fertilization or in presmolt growth periods from February to April in 1996 and 1997. Smolting rates were high (range 93.5–95.3%) and the incidence of deformities was low (< 1%) in both groups. Marine survival was lower in the triploid group, largely as a consequence of higher losses sustained during a period of chronic stress, when triploid losses were 9% higher. Growth patterns were similar for the first 11 months in sea water. Although graded triploid salmon were heavier in January 1998 (AF3N 1.62 ± 0.033 kg, AF2N 1.46 ± 0.36 kg, P < 0.05), when the fish were harvested in May 1998 diploid salmon were significantly heavier than triploid salmon although there was no significant difference in weights after evisceration (AF3N 2.40 kg ± 0.04 AF2N 2.49 kg ±0.03). The increase in weight of the diploids between winter and harvest reflects the growth spurt that occurs in maturing fish in the spring. Overall yields of triploid salmon in salt water were lower as a result of inferior survival.     

Comparative performance of growth,vertebral structure and muscle composition in diploid and triploid Paralichthys olivaceus

Growth, skeletal structure and muscle composition of cold‐shock‐induced triploid olive flounder Paralichthys olivaceus were investigated. The average values of total length and total weight of triploids were higher than those of diploids from 5 to 11 months posthatch (mph). The growth difference disappeared after 11 mph. The skeletal structure of flounder at 11 mph was observed by X‐ray imaging method. There are four kinds of vertebral deformity including vertebrae fusion, one‐sided compression, two‐sided compression and vertically shifted. The trunk region (V8–18) and tailing end of the vertebral column were the predominant locations of deformity. In general, the frequencies of vertebral deformities in triploids (60.0%) were higher than those in diploids (33.3%, p < 0.05). Both the number of fish with deformed vertebrae and the average frequencies of deformed vertebrae in triploids were significantly higher than those in diploids (p < 0.05). The muscle tissues of diploid and triploid flounder at 11 mph contain the same types of fatty acid and free amino acid profiles. The number of fatty acids with significant higher contents in diploids and triploids was one and ten, respectively (p < 0.05). The contents of free amino acids showed no difference between triploid and diploid fish.     

Analysis of growth, weight and relevant indices of diploid and triploid population of tench Tinca tinca (Linnaeus 1758)

This study determined biometric and weight parameters and relevant indices of diploid and triploid tench. Altogether, 137 siblings of tench were studied. The effect of ploidy level appeared in significantly better growth of triploids (P<0.001) as to biometric [total length (TL), standard length (SL), body height (BH), body width (BW)] and weight [fish weight (FW), carcass weight (CW)] parameters of T₃ of both sexes and of T₃₊ females. The effect of ploidy level also appeared as significantly higher dressing percentage (DP; P<0.001) of triploid T₃ females compared with other groups, significantly higher gonad weight (GW) and gonadosomatic index (GSI; P<0.001) of diploid T₃ females, as well as GSI and hepatosomatic index of diploid T₃₊ females. The effect of sex appeared in significantly higher (P<0.001) biometric (TL, SL, BH, BW) and weight (FW, CW) parameters of T₃ females of both ploidy levels, as well as of triploid T₃₊ females. The effect of sex also appeared as significantly higher DP (P<0.001) of males in diploid T₃ fish, as well as of males of both ploidy levels in T₃₊ fish and significantly higher GW and GSI (P<0.001) of females in diploid T₃ fish, as well as of females of both ploidy levels in T₃₊ fish. This study shows evidence for faster somatic growth and bigger final weight of triploid populations of tench compared with diploids in both age categories T₃ and T₃₊.     

Growth enhancement following dietary delivery of recombinant porcine somatotropin to diploid and triploid coho salmon (Oncorhynchus kisutch)

Diploid (2n) and triploid (3n) coho salmon were fed upon control (diet 1), recombinant porcine somatotropin (rpST)-supplemented (diet 2; 20 μg rpST/g body wt/day) or rpST/antacid/detergent-containing feed (diet 3; 20, 100 &; 20 μg/g body wt/day respectively) for 16 wk. Fish were weighed and measured bimonthly and their relative performances assessed. By wk 4, 2n and 3n groups fed upon diet 2 were significantly heavier and longer than control 2n fish. At the termination of the trial, diet 3 fed animals were greater in weight than all other treatment groups. Diet 3 salmon also returned better feed conversion efficiencies than either diet 1 or 2 groups. RpST therapy induced a 28.7% and 60.2% increase in group wt for 3n diet 2/3 coho versus (?vs?) controls. Likewise, 2n rpST-treated fish increased wt ?vs?2n control coho by 17% and 50% respectively over the same period. No differences were recorded between groups for body moisture, but diet 2, 3n and both diet 3 groups exhibited decreased condition factors when compared to control fish (p ≤ 0.05).