Expression profiles of sex-related genes in gonads of genetic male Takifugu rubripes after 17β-estradiol immersion

Estradiol treatment during early life stages of tiger puffer Takifugu rubripes induces feminization in genetic males. However, the ovaries in genetic males may revert to testes once estradiol treatment is halted.Therefore studies should investigate molecular mechanisms underlying ovary-to-testis recovery in genetic males after treatment. In the present study, tiger puffer were exposed to 10, and 100 μg/L 17 p-estradiol(E_2)from 15 to 100 days post-hatching(dph), then gonad phenotypes and expression profiles of six sex-related genes(cyp19a,foxl2, dmrtl, amh, sox9a, and sox9b) were characterized after the exposure. Results showed that both 10 and 100 μg/L E_2 induced ovarian development in genetic males at 100 dph. However, all ovaries induced by 10 μg/L E_2 first developed into intersexual gonads and subsequently reverted to testes after the exposure. As for treatment of 100 pg/L E_2, while the rest of the ovaries maintained morphological stability,percentages of intersexual gonads reached 38%-57%, and none were reverted to testes. Increased mRNA levels of cyp19 a,foxl2 and sox9b and decreased mRNA levels of dmrtl, amh, and sox9a were observed during the ovarian development in genetic males. While contrary gene expression profiles were detected during ovary-to-testis transformation. The mRNA levels of all the six genes were increased during the development of intersexual gonads. These results indicated that up-regulation of dmrtl, amh and sox9a is associated with initial ovary-to-intersexual transformation, and suppression of foxl2, cypl9a and sox9b is essential for complete ovary-to-testis recovery in genetic males. This research will help to trace the molecular processes underlying gonadal transformation in teleosts.     

Characterization of Bacterial Community Associated with Four Organs of the Yesso Scallop (Patinopecten yessoensis) by High-Throughput Sequencing

We used Illumina high-throughput sequencing of PCR-amplified V3-V4 16 S rRNA gene regions to characterize bacterial communities associated with the adductor muscles, gills, gonads and intestines of the Yesso scallop(Patinopecten yessoensis) from waters around Zhangzidao, Dalian, China. Overall, 421,276 optimized reads were classified as 25 described bacterial phyla and 308 genera. Firmicutes, Proteobacteria, Tenericutes, Bacteroidetes, Chlamydiae and Spirochaetae accounted for 97% of the total reads in the four organs. The bacterial 16 S rDNA sequences assigned to Firmicutes and Proteobacteria were abundant in the adductor muscles, gills and gonads; while reads from Tenericutes were dominant in the intestines, followed by those from Firmicutes, Chlamydiae, Proteobacteria and Bacteroidetes. At the genus level, the dominant genera in the adductor muscles, gills and gonads appeared to be Bacillus, Enterococcus and Lactococcus, whereas Mycoplasma was dominant in the intestines. The relative abundances of Bacillus, Enterococcus, Lactococcus, Alkaliphilus, Raoultella, Paenibacillus and Oceanobacillus were significantly lower in the intestine than in the other three organs. Cluster analysis and principal coordinates analysis of the operational taxonomy units profile revealed significant differences in the bacterial community structure between the intestine and the other three organs. Taken together, these results suggest that scallops have intestine-specific bacterial communities and the adductor muscles, gills and gonads harbor similar communities. The difference in the bacterial community between organs may relate to unique habitats, surroundings, diet and their respective physiological functions.     

环沟格特哈(Katelysia rimularis)的生殖周期

周年采集环沟格特蛤（Katelysiarimularis）分别测定生长指标（总重、软体部重、性腺重、肝重和软体部干重等）并用组织切片观察性腺发育的状况，以确定性腺发育的周期和时间。组织的光镜研究说明环沟格特蛤雌雄性腺发育均可分为增殖期、生长期、成熟期、排放期和休止期。在厦门吴冠镇环沟格特蛤1月开始其性腺发育,3月进入生长期，5～6月成熟，5～11月都有产卵，每年7月11月各有一次排卵高峰期。     

Higher Dietary Arachidonic Acid Levels Improved the Growth Performance,Gonad Development,Nutritional Value,and Antioxidant Enzyme Activities of Adult Sea Urchin(Strongylocentrotus intermedius)

The gonads of sea urchins(Strongylocentrotus intermedius) are characterized by high levels of arachidonic acid(ARA, 20:4 n-6) and eicosapentaenoic acid(EPA, 20:5 n-3). However, to our knowledge, little information is available regarding the physiological response of adult sea urchins to dietary ARA. In the present study, four dietary feeds were formulated with graded ARA(0, 0.5%, 1%, and 2% dry diet). Each diet was randomly allocated to three cages during a 56-day feeding experiment. The results showed that the sea urchin weight gain rate(WGR) and the gonadosomatic index(GI) significantly increased as ARA was equal to or above 1.0% of dry diet(P 0.05). The activities of superoxide dismutase(SOD), catalase(CAT), and total anti-oxidative capacity(T-AOC) were the highest in the coelomic fluid of sea urchins that were fed diets with 1% ARA. The total essential amino acid(TEAA) and its ratio to total non-essential amino acid(TNEAA) showed a similar tendency to WGR and GI as dietary ARA increased, and the highest TEAA and TEAA/TNEAA were observed in the gonads of sea urchins that were fed diets with 1% ARA. Levels of ARA and ARA/EPA of the gonads increased while n-3/n-6 polyunsaturated fatty acid(PUFA) decreased with the increase of dietary ARA(P 0.05). EPA in the gonads of experimental animals fed with formulated feeds showed no significant differences(P 0.05), but was significantly lower than those fed with kelp(P 0.05). These results suggested that relatively higher levels of ARA(1% dry diet) significantly promoted growth, gonad development, activities of antioxidant enzymes, as well as nutritional values(TEAA, TEAA/TNEAA, and PUFA) of adult S. intermedius.     

鞍带石斑鱼人工繁殖及胚胎发育研究

为建立鞍带石斑鱼(Epinephelus lanceolatus)人工繁殖技术,研究了鞍带石斑鱼亲鱼培育、催产及胚胎发育过程。根据性腺检查及催产结果,判定鞍带石斑鱼在广东的繁殖季节为每年的7-9月份,产卵水温在28~31℃。使用10~12尾雌鱼和3尾雄鱼搭配进行催产实验,亲鱼未出现发情和追尾现象,虽能产卵但均未受精,通过人工授精可得受精卵。对鞍带石斑鱼胚胎发育进行了观察,详细描述从受精卵到初孵仔鱼的28个具体发育时期的形态特征和发育时间,结果表明,将鞍带石斑鱼胚胎发育划分为卵裂期、囊胚期、原肠胚期、神经胚期和器官形成期。在水温(27±0.5)℃、盐度31.0、pH 7.8的海水中,鞍带石斑鱼胚胎历时25 h 40 min完成整个胚胎发育孵化出膜。     

Studies on the eggs and larvae,ofSardinella aurita (Cuv. & Val.) in the fishing grounds of Southern Fujian and the Taiwan Bank

In this paper, the morphological characters of eggs and larvae ofSardinella aurita (Cuvier & Valenciennes), its spawning ground, spawning seasons and spawning conditions have been studied. Sardinella aurita likes to breed in the upwelling area and selects the Waixie fishing ground as its main spawning ground. The months from February to September are its spawning seasons, reaching its peak in April. In the main spawning ground, the temperature of the surface layer was found to be 24.4–25.2°C, the salinity 33.87–34.07%. and the depth of water between 34–60m. The distribution of the larvae is closely related to the distribution of plankton, the path of migration of adult fish, and the current direction of the water system. In order to protect fishery resources, it is necessary to prohibit catching the spawning fish in the Waixie fishing ground in April, and the catching of immatures from March to June. This paper was published in Chinese inOcean. Limn. Sinica 14(3): 240–248, 1983.     

Microsatellite-Centromere Mapping in Japanese Scallop(Patinopecten yessoensis) Through Half-Tetrad Analysis in Gynogenetic Diploid Families

Gene-centromere mapping is an essential prerequisite for understanding the composition and structure of genomes. Half-tetrad analysis is a powerful tool for mapping genes and understanding chromosomal behavior during meiosis. The Japanese scallop(Patinopecten yessoensis), a cold-tolerant species inhabiting the northwestern Pacific coast, is a commercially important marine bivalve in Asian countries. In this study, inheritance of 32 informative microsatellite loci was examined in 70-h D-shaped larvae of three induced meiogynogenetic diploid families of P. yessoensis for centromere mapping using half-tetrad analysis. The ratio of gynogenetic diploids was proven to be 100%, 100% and 96% in the three families, respectively. Inheritance analysis in the control crosses showed that 51 of the 53 genotypic ratios observed were in accordance with Mendelian expectations at the 5% level after Bonferroni correction. Seven of the 32 microsatellite loci showed the existence of null alleles in control crosses. The second division segregation frequency(y) of the microsatellite loci ranged from 0.07 to 0.85 with a mean of 0.38, suggesting the existence of positive interference after a single chiasma formation in some chromosomes in the scallop. Microsatellite-centromere distances ranged from 4 c M to 42 c M under the assumption of complete interference. Information on the positions of centromeres in relation to the microsatellite loci will represent a contribution towards the assembly of genetic maps in the commercially important scallop species.     

Comparison of growth and survival of larvae among different shell color stocks of bay scallop Argopecten irradians irradians (Lamarck 1819)

In order to improve production and breed new broods of bay scallopArgopecten irradians irradians, different-colored orange, purple and white lines were established by two mating methods of self-fertilization and mass spawning at a commercial scallop hatchery in spring, 2002. And then larval growth and survival of different lines was compared to test whether there is a relationship between shell color and growth and survival at early developmental stage. Both growth and survival have no significant differences among different experimental larvae of self-fertilization or mass spawning. Results are as following in the order of orange, purple and white shell stock. For the self-fertilization, growth rates of larvae were 6.174, 6.412, and 6.599 μm/d, respectively. Survival rates of larvae at Day 3 were 74.41%, 76.86%, and 82.05%; Day 6 were 49.14%, 65.63%, and 52.79%; and Day 9 were 25.06%, 20.80%, and 26.47%, respectively. For the mass spawning, the growth rates were 7.836, 7.941, and 7.878 μm/d, respectively. Survival rates at Day 3 were 93.05%, 91.95%, and 92.50%; Day 6 were 79.17%, 78.05%, and 82.50%; and Day 9 were 34.72%, 36.67%, and 38.33%, respectively. The absence of any relationship between shell color and growth and survival at the larval stage may be resulted from their common genetic basis. This work was financially supported by Project of Scientific Innovation, Chinese Academy of Sciences (ZKCX2-211).     

ARTIFICIAL CULTURE OF AMPHIOXUS (BRANCHIOSTOMA BELCHERI TSINGTAUENSE)

Mature amphioxus (Branchiostoma belcheri tsingtauense) collected by trawl in June, 1994 from Qingdao Shazikou sea area bottom sands, were cultured under controlled indoor conditions, where they spawned and their eggs were fertilized and hatched into larvae which gradually settled at the bottom and dug into the sands after 40-50 days culture in a water trough outdoors. The total survival rate of the larvae(LSR)was 5.5% before they went into the sands, 1.9% in 4 months, 0.7% in 10 months. The survival rate of the young fish which had dug into the sands (YSR) was 35.6% in 4 months, and 12.6% in 10 months. The amphioxus number tended to be constant from the 11th month on. In the first five months after incubation, the amphioxus body length increased by an average of about 1.5 mm a month, and about 0.4 mm a month from Dec. to May of next year. The maximum length after ten months was 24 mm; the average was about 11 mm. After two years culture in the water trough, the maximum length could reach 34-35 mm, when the amphioxus gonads began to develop.     

Effect of Long-Term Low-Salinity Culture on the Survival,Growth, and Nutrient Composition of Mud Crab Scylla paramamosain

This study investigated the effects of long-term low-salinity on the growth performance,proximate composition,amino acid and fatty acid profiles of Scylla paramamosain.The salinity of the low-salinity-culture group(LC)and maintained-salinity-cul-ture group(MC)were set at 5 and 25 respectively.After rearing for 30 d,the survival rate(SR)of the LC group was significantly lower,whereas the average daily molting frequency(ADMF)was significantly higher than those in MC group(P<0.05).There were no significant difference for weight gain rate(WGR)and specific growth rate(SGR)between the two groups.The moisture content of the LC group was slightly higher than the MC group without significant difference.The contents of ash,crude lipid and crude pro-tein in the LC group were lower than those in the MC group,but no significant differences were found except for ash content.A total of 17 free amino acids and 23 fatty acids were identified from the muscle of S.paramamosain.The total amino acids(TAA),essen-tial amino acids(EAA)and essential amino acid index(EAAI)had no significant difference between the two groups.The crabs from LC group had a significantly higher HUFA,including EPA,DHA,ARA.The percentage of n-3 PUFA and the ratio of n-3/n-6 PUFA were significantly higher in the LC group.These results indicated that S.paramamosain rearing at low salinity had significantly de-creased SR;however,the low-salinity environment significantly promoted the accumulation of unsaturated fatty acids(UFAs),espe-cially the n-3 PUFA,which might be related to the osmoregulation mechanism.     

凤鲚、湖鲚和刀鲚卵巢氨基酸、脂肪酸及矿物元素分析

对长江下游凤鲚、湖鲚和刀鲚卵巢氨基酸、脂肪酸和矿物元素组成及含量进行了测定分析。结果表明:三种鲚属鱼类卵巢鲜样中总氨基酸、必需氨基酸、呈味氨基酸含量差异不显著;必需氨基酸含量(w)为5.36%～6.32%,占总氨基酸含量为:0.43%、0.41%和0.43%,均以亮氨酸含量最高而色氨酸最低,赖氨酸含量(w)为0.96%～1.04%;饱和脂肪酸、单不饱和脂肪酸和多不饱和脂肪酸差异较显著,脂肪酸组成均以油酸含量最高,EPA和DHA总量(w)为8.00%～11.81%;矿物质含量大多也存在显著差异,常量元素含量最高的为钾(16970～22220mg/kg),微量元素则为锌(262.0～277.0mg/kg);不同水域生长环境对鲚属鱼类卵巢氨基酸含量影响不显著,而对脂肪酸和矿物质含量影响较显著。     

Inheritance Pattern of Microsatellite Loci and Their Use for Kinship Analysis in the Japanese Scallop Patinopecten yessoensis

The inheritance mode of seven microsatellite markers was investigated in Patinopecten yessoensis larvae from four con-trolled crosses,and the feasibility of using these markers for kinship estimation was also examined. All the seven microsatellite loci were compatible with Mendelian inheritance. Neither sex-linked barriers to transmission nor major barriers to fertilization between gametes from the parents were evident. Two of the seven loci showed the presence of null alleles in two families,suggesting the...     

Advances in the Study on Nutrient Requirements of Grouper（Epinephelus sp. ）：a Review

The paper reviews the recent advances in studying grouper nutrition requirement for the development of cost-effective and environmentally friendly artificial diets. It consists of seven parts: protein and amino acid, lipid and essential fatty acid, carbohydrate, vitamin, mineral, alternative protein source, broodstock and larval nutrition. The review provides some basic information for further investigation of nutrient requirements of groupers.     

Triploidy induction with heat shocks toPenaeus chinensis and their effects on gonad development

Heat shocks effectively produced triploids inPenaeus chinensis. Fertilized eggs heat shocked (28–32°C) for 8 to 16 minutes, starting from 8 to 20 minutes after spawning, resulted in triploidy induction rates of 39%–75%. Several triploid populations were cultured to 10 cm. In a triploid population, two kinds of ovaries were observed. Histological examination showed apparent differences between these two kinds of ovaries, whereas among male shrimp, there were no such differences. Contribution No. 3465 from the Institute of Oceanology. Chinese Academy of Sciences. The study was supported by the International Foundation for Science, Asian Fisheries Society and Chinese Climbing Project PDB 6–2, and Chinese National 863 Project (No. 863-819-01-05).     

Advances in the Study on Nutrient Requirements of Grouper (Epinephelus sp. ): a Review

The paper reviews the recent advances in studying grouper nutrition requirement for the development of cost-effective and environmentally friendly artificial diets. It consists of seven parts: protein and amino acid, lipid and essential fatty acid, carbohydrate, vitamin, mineral, alternative protein source, broodstock and larval nutrition. The review provides some basic information for further investigation of nutrient requirements of groupers.     

Effects of Sodium Nitrate and Sodium Acetate Concentrations on the Growth and Fatty Acid Composition of Brachiomonas submarina

1　Introduction　Microalgae ,asthefirstlinkofmarinefoodchain ,canbeusedfortheaccumulationofpolyunsaturatedfattyacids (PUFAs) ,whichhavebeenshowntobeofmajorimportancenotonlyinthe preventionofhu man’sheartandcirculatorydiseases (DyerbergandJorgensen ,1982 ;Marcus ,1985 )butalsoinaquacul tureasincipientfeed (Sakamotoetal.,1998) .How ever ,thePUFA productivitiesbymicroalgaearelowincomparisonwithbacteriaandfungi.TherearetwopossiblewaysinpromotingthePUFA productionofmicroalgae.Oneistoobtaint…     

繁殖季节彩虹鲨卵巢发育的组织学研究

对繁殖季节彩虹鲨卵巢的结构进行显微观察。结果表明,繁殖季节彩虹鲨卵巢,大多处于Ⅳ期和Ⅴ期,少数处于Ⅲ期。在Ⅳ和Ⅴ期卵巢中,Ⅱ～Ⅵ时相的卵母细胞同时存在,说明彩虹鲨为分批产卵类型。Ⅱ时相卵母细胞出现卵黄核,Ⅲ时相卵母细胞出现放射膜和皮质液泡,Ⅳ时相卵母细胞的受精孔和精孔细胞明显。     

Effect of temperature and photoperiod on maturation of Cephalothrix hongkongiensis （Nemertea： Palaeonemertea）

The limited knowledge about the relationships between environmental factors and the maturation and reproductive seasonality of nemerteans is mostly obtained from field studies, and in many cases is conjectural. Few species have been studied under well-controlled laboratory conditions. In this work, the effects of temperature and photoperiod on gonad development and spawning of Cephalothrix hongkongiensis were observed by rearing worms under different temperatures and photoperiods. Worms reared at 4℃ and 6℃ showed no growth or gonadal development, and all died within 109 days. At 8℃ and 12℃, gonads could be seen from 90 days to the termination of the 360-day experiment, but no spawning was observed. Worms maintained at 16, 24, and 28℃ showed both gonadal development and spawning, and matured faster at higher temperatures. Under a fixed temperature(16℃), photoperiod did not have any significant impact on maturation and spawning. It is likely that temperature plays a major role in regulating the maturation and reproductive cycle of C. hongkongiensis.     

Breeding and mass scale rearing of clownfish Amphiprion percula: feeding and rearing in brackishwater

Breeding and mass scale larval rearing of clownfish Amphiprion percula is very limited in brackishwater. We designed an indoor program of A. percula culture in brackishwater with a salinity of 24±1, during which the impacts of feed type, water temperature, and light intensity, on the efficiency of its reproduction, were revealed. The fish were accommodated along with sea anemones in fibre glass tanks to determine the influence of brooder diet on breeding efficiency. Higher reproductive efficiency [number of eggs laid (276 22.3 eggs)] was observed when fish were fed live Acetes sp. rather than clam (204 16.4 eggs), trash fish (155 12 eggs) and formulated feed (110 10 eggs). The spawning rate was increased during September and October (water temperature, 28.74 0.55°C) on average of 2.4 spawning per month; and low spawning rate was in January (water temperature, 24.55 0.45°C) on average of 1 spawning per month. Among three light intensities (100, 500, and 900 lx) set to evaluate larval survival rate, larvae showed the highest survival rate (65.5%) at 900 lx. The breeding method specifically in brackishwater developed in the present study is a new approach, will help the people from the regions of estuary and backwater to enhance their livelihood and it will lead to reduce the exploitation from the wild habitat.     

方斑东风螺不同批次卵质量的比较

2005年5月~8月在雷州市覃斗镇海威水产种苗基地的室内水池进行方斑东风螺亲螺培育,对亲螺不同批次产出的卵囊、卵、及其幼虫进行比较。结果表明:随着亲螺产卵批次的增加,亲螺产卵囊数、卵囊含卵数、产卵量逐次减少;卵囊长、宽逐次变小,厚逐次增加,形状由葵花子片状的比例大变为棒状的比例大;幼虫浮游期逐次延长;变态期幼虫壳长逐次变小;各批次之间的变态率逐次降低;卵径、孵化率、出囊幼虫大小差异不显著。