Chromosome behavior of heat shock induced triploid in Fenneropenaeus chinensis

Triploidy was induced in Chinese shrimpFenneropenaeus chinensis by 30±0.5°C heat shock treatment (initiated at 20 min after fertilization) for 10 min to inhibit the release of PB2 at 18.0°C. The highest triploid rate obtained was 84.5% in nauplius stage. The effect of heat shock treatment on meiosis and cleavage of eggs was investigated in this work aimed to establish efficient procedures for triploid induction and to gain understanding of the mechanism of triploid production. Three pronuclei that could be observed in the treated eggs under fluorescence microscope developed into triploid embryos. Some abnormal chromosome behavior was observed in heat shocked eggs. Contribution No. 4407 from the Institute of Oceanology, Chinese Academy of Sciences. This work was supported by Project under Major State Basic Development Program (G1999012009) and Knowledge Innovation Program of the Chinese Academy of Sciences (KZCX 2-211)     

Polyploidy induction by hydrostatic pressure shock and embryo development of sea cucumber Apostichopus japonicus

The manipulation of the chromosome set for commercially valuable marine animals is important for enhancing aquacultural production. In this study, triploid and tetraploid sea cucumber Apostichopus japonicus were induced by hydrostatic pressure shock, and the conditions of appropriate induction were tested with different starting times, and hydrostatic pressure intensities and durations. The highest rate of triploid induction reached 20% and that of tetraploid was 60%. In consideration of the survival rate and hatch rate, the appropriate treatment for triploid was 55 Mpa of hydrostatic pressure for 5 min at 55 min after fertilization (a.f.), while for tetraploid it was 60 Mpa for 5 min at 61 min a.f. The triploid of the sea cucumber could survive through the pelagic larval stage and attachment stage, and develop like the control group of the experiment. The tetraploid, however, could not survive the attachment stage.     

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).     

珠江斑鱯的早期胚胎发育观察

观察了珠江斑鳠3批次受精卵的胚胎发育,描述及记录了胚胎发育各期的形态特征及所需时间。斑鳠的成熟卵子呈圆形,黄色,卵子直径为3.1~3.8 mm,吸水后最大卵径为4.0~4.2 mm,受精卵黏性很小。珠江斑鳠胚胎发育与其他硬骨鱼类基本相似,属于盘状卵裂。在水温(27.5±0.5)℃时,受精后约1 h 10 min进入胚盘形成期,受精2 h 15 min后进入卵裂期,受精后7 h 50 min,进入囊胚期,再经约13 h 45 min进入原肠期,受精后21 h15 min,胚孔封闭,当受精39 h 5 min后,心脏开始出现搏动,受精后55 h,仔鱼开始孵出。     

Artificial induction of mito-gynogenetic diploids in large yellow croaker (Pseudosciaena crocea) by hydrostatic pressure

The present study investigated conditions for inducing mito-gynogenetic (endomitosis) diploids by hydrostatic pressure in the large yellow croaker Pseudosciaena crocea. In haploid control groups, the development of eggs was activated with ultraviolet radiated semen. All fry presented typical haploid syndrome in the haploid control groups, and were verified as haploids using cytometry. After hydrostatic pressure treatment, morphologically normal fry reappeared at different frequencies according to the intensity and time of pressure shock. Fry with normal appearance in the pressure treated groups were verified as gynogenetic double haploids (GDHs), containing only one allele from the female parent at all four diagnostic microsatellite loci. For a fixed duration of 3 min, the optimal intensity of blocking the first mitosis was determined to be 40 Mpa, which was similar to that of blocking the second meiosis. There was a “window” of starting time, from 36.1 min to 38.1 min post-insemination at 25.0±1.0°C, within which the production of GDHs was not significantly different. Maximum production of morphologically normal fries, 9.36%±2.97% of developed eggs, was found when the eggs were shocked with hydrostatic pressure at 40 Mpa for 3 min, starting from 38.1 min post insemination at 25.0±1.0°C.     

STUDY ON EMBRYONIC DEVELOPMENT AND EARLY GROWTH OF TRIPLOID AND GYNOGENETIC DIPLOID LEFI—EYED FLOUNDER, PARALICHTHYS OLIVACEUS （T. et S. ）

The early effects of chromosomal manipulation of eggs and sperm on the yields of trip-loid and gynogenefic diploid larvae of Paralichthys olivaceus were investigated. Triploidy was achieved by cold shocking fen.ilized eggs at 0 - 2℃ for 45 minutes duration 5 minutes after fen.ilization, and the in-duced triploidy rates were 31.2% - 50% and the relative hatching rates were 53.3% - 99%. Gynnge-aetic diploids were obtained when eggs were inseminated with irradiated sperm and cold shocked at 0 -2℃ for 45 minutes duration 5 minutes after fertihzation. The induced gynogenetic diploid rates and the relative hatching rates were 94 % - 96 % and 48.5 % - 68.5 % respectively. The embryonic development of the triploid experimental group and of the gynogenetic diploid experimental group was delayed at first compared with the control group. But from the gastrula stage, it was not delayed anymore. There were no significant differences in the growth of the triploid experimental group larvae and the control group larvae, and in the growth of the gynogenetic diploid experimental group larvae and the control group larvae ac-cording to Student‘s t-test (α = 0.05). The relationship between the early growth of the triploid experi-mental group larvae and that of gynogenetic diploid experimental group larvae was also studied.     

不同冷却时间对高温花岗岩可钻性实验研究

为了研究高温后花岗岩的可钻性和微观损伤变化, 同时也为了研究高温后不同冷却时间对可钻性的影响, 对高温热处理后的花岗岩冷却不同时间(2, 4, 24, 48 h), 通过可钻性实验和铸体薄片鉴定, 得到高温对花岗岩可钻性的影响规律和影响机理, 同时也得到不同冷却时间对花岗岩可钻性的影响。研究结果表明, 花岗岩在热处理不超过500℃和自然冷却2 h的约束下始终保持一个较高的可钻性级值, 在冷却4, 24, 48 h后, 高温对可钻性的影响表现为3个阶段(第一次劣化阶段、强化阶段、第二次劣化阶段)。微裂纹产生的位置及数量影响着岩石抵抗破碎的难易程度, 400℃热处理后花岗岩内部微裂纹开始显著增加, 当石英颗粒内部产生大量微裂纹时, 花岗岩的可钻性显著降低。100℃热处理后同时冷却不超过4 h会显著影响花岗岩的可钻性, 200~400℃热处理后, 花岗岩的可钻性级值会随着冷却时间(4~48 h)的继续增加显著增加, 500℃对花岗岩产生的损伤是不可恢复的, 600℃已经完全使花岗岩劣化。弄清高温和冷却时间对花岗岩可钻性的影响, 可以为干热岩资源的高效开采提供基础的理论支撑。      

Cytogenetic mechanism for the aneuploidy and mosaicism found in tetraploid Pacific oyster Crassostrea gigas (Thunberg)

Chromosome constitution was investigated in adult tetraploid Pacific oyster produced by blocking the first polar body of triploid eggs which were fertilized with haploid sperms. A high incidence of aneuploid and heteroploid mosaics were found among the offspring. Of 20 individuals identified, only 9 (45%) were eutetraploid which contained 40 chromosomes; 2 (10%) were aneuploid (hypotetraploid), which contained 39 and 38 chromosomes, respectively; and 9 (45%) were heteroploid mosaics. One mosaic was consisted of cells containing 40 and 39 chromosomes, respectiovely (1:1 in cell number), while the other 8 were consisted of cells containing chromosomes varying between tetraploid and triploid. It was also interesting to note that 3 mosaics even contained some diploid cells with 20 chromosomes. A certain number of cells of 2 tetraploids and 8 mosaics spread with 32–37 well-scattered and some clumped chromosomes at metaphase. The percentage of aneuploid cells with chromosomes varying between triploid and tetraploid correlated significantly with that of heteroploid mosaics cells with clumping chromosomes (P<0.05). Our findings suggested that reversion existed in both tetraploid and triploid oyster and chromosome clumping may underline the chromosome elimination in tetraploid oyster. It seems that the reversing cells, at least some of them, continuously eliminate their chromosomes until the most stable diploid state is established.     

Triploidy induction in Pacific oysterCrassostrea gigas by caffeine with thermal shock

Zygotes of Pacific oyster (Crassostrea gigas) were treated for triploid induction with caffeine (10 mmol/L, 15mmol/L and 20mmol/L) in combination with thermal shocks (at 40 minute post-fertilization) lasting for 5 and 10 minutes. The highest yield of triploids, 41.5%, was obtained from the treatment with 20 mmol/L caffeine at 34°C lasting 10 minutes. The triploid levels were less than 30.0% in other treatments. Triploid induction was more effective in treatment with 15–20 mmol/L caffeine at 34–38 °C than with lower concentrations of caffeine at temperatures below 32 °C. Our results suggest that triploid induction with caffeine in combination with thermal shocks is less efficient than some other methods reported previously.     

苏氏圆腹胚胎发育的初步观察

观察两批苏氏圆腹受精卵的胚胎发育,描述并记录胚胎发育各期的形态特征及发育时间。结果表明:苏氏圆腹成熟卵呈圆球形,淡黄色,富有光泽,卵黄分布均匀,相对密度略大于水,卵径0.93～1.11 mm,平均0.98±0.19 mm。受精卵吸水后呈微黏性,属黏性卵,卵膜略膨胀,卵径0.99～1.15 mm,平均1.08±0.27 mm。苏氏圆腹胚胎发育属于盘状卵裂,与其他淡水硬骨鱼类基本相似。其发育过程可分为:受精卵、卵裂期、囊胚期、原肠期、神经胚期、器官分化期、出膜期和初孵仔鱼。在水温26±1℃的条件下,苏氏圆腹胚胎孵化历时24 h 45 min,总积温640.90 h.℃。初孵仔鱼体全长3.20～3.60 mm。     

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。     

Reduction of hydrocarbon contamination on viability of Acartia pacifica benthic resting eggs

The potential effect of hydrocarbon contamination on the hatching success of benthic resting eggs of Acartia pacifica in Xiamen Bay was investigated experimentally. The number of nauplii emerging from the sediment samples decreased with increasing Fuel Oil #0 concentration. The estimated rate of mortality increased markedly with the increase of Fuel Oil #0 concentration. Successive fuel Oil #0 concentrations from 50 mg/kg to 5000 mg/kg reduced the number of hatched nauplii by 3.8% -100%. The mortality of A. pacifica resting eggs due to Fuel Oil #0 contamination did not significantly increase as time progressed at each concentration level. The LC50 values of resting eggs, changing from 237.12 to 279.59 mg/kg, remained at an almost stable level in two months. The number of A. pacifica nauplii that hatched from the sediment at 10℃ was higher than those from the sediment at 30℃, which indicates that the toxicity of Fuel Oil #0 on A. pacifica resting eggs increases with increasing temperature.     

不同温度下马氏珠母贝干露耐受能力及其免疫相关基因表达的变化

【目的】探究马氏珠母贝(Pinctada fucata martensii)对干露胁迫的响应,为马氏珠母贝健康养殖和遗传育种提供基础资料。【方法】在不同温度下对马氏珠母贝进行干露胁迫,探究温度对马氏珠母贝干露耐受的影响;筛选出马氏珠母贝干露胁迫敏感组和耐受组,比较两组的免疫相关基因表达量和植核后休养期存活率。【结果】(1)22~34℃条件下,马氏珠母贝干露耐受能力随温度升高及干露时间延长而降低;(2)22℃下,S组和T组的脯氨酸羟化酶(PHD)、葡萄糖转运蛋白(GLUT)、髓样分化因子(MYD88)、热休克蛋白(Hsp70、Hsp20)和凋亡抑制因子(cIAP)6个基因在干露过程中表达量基本呈上升趋势,组间差异显著(P<0.05);(3)植核试验结果表明耐受组植核后休养期的存活率显著高于敏感组(P<0.05)。【结论】马氏珠母贝在高温环境下干露耐受能力降低,在适温条件下耐受组比敏感组表现出较高的存活率。     

Isolation and characteristics of one marine psychrotrophic cellulase-generating bacterium Ar/w/b/75°/10/5 from Chuckchi Sea,Arctic

Microorganisms living in polar zones play an important part as the potential source of organic activity materials with low temperature characteristics in the bio-technological applications. A psychrotrophic bacterium (strain Ar/w/b/75°/10/5) , producing cellulose at low temperatures during late-exponential and early-stationary phases of cell growth, was isolated from sea ice-covered surface water in Chuckchi Sea, Arctic. This bacterium, with rod cells, was Gram-negative, slightly halophilic. Colony growing on agar plate was in black. Optimum growth temperature was 15℃. No cell growth was observed at 351 or above. Optimum salt concentration for cell growth was between 2 and 3 % of sodium chloride in media. Maximal cellulase activity was detected at a temperature of 35℃ and pH8. Cellulase was irreversibly inactivated when incubated at 55℃ within 30 min. Enzyme can be kept stable at the temperature no higher than 25℃. Of special interest was that this bacterium produced various extracellular enzymes i     

Expression profiles of two small heat shock proteins and antioxidant enzyme activity in Mytilus galloprovincialis exposed to cadmium at environmentally relevant concentrations

Small heat shock proteins encompass a widespread but diverse class of proteins,which play key roles in protecting organisms from various stressors.In the present study,the full-length cDNAs of two small heat shock proteins(MgsHSP22 and MgsHSP24.1)were cloned from Mytilus galloprovincialis,which encoded peptides of 181 and 247 amino acids,respectively.Both MgsHSP22 and MgsHSP24.1 were detected in all tissues examined by real-time PCR,with the highest expression being observed in muscle and gonad tissues.The real-time PCR results revealed that Cd signifi cantly inhibited MgsHSP22 expression at 24 h and MgsHSP24.1 at 24 and 48 h under 5 μg/L Cd 2 + exposure.MgsHSP24.1 expression was also signifi cantly inhibited after 50 μg/L Cd 2+ exposure for 48 h.With regard to antioxidant enzymes,increased GPx and CAT activity were detected under Cd 2+ stress(5 and 50 μg/L),while no signifi cant difference in SOD activity was observed throughout the experiment.Overall,both MgsHsps and antioxidant enzymes revealed their potential as Cd stress biomarkers in M.galloprovincialis.     

Heat-shock protein 70 expression in shrimp Fenneropenaeus chinensis during thermal and immune-challenged stress

Using western immunoblotting we obtained heat-shock protein 70 (HSP70) induction data and distribution in different tissues from shrimpFenneropenaeus chinensis during thermal and immune-challenged stresses. This is probably the first report of the effects of various stressors on the expression of HSP70 in shrimp. HSP70 was prominently induced in hepatopancreas and gills, but not in muscle, eyestalk and hemolymph, when the shrimp were exposed to heat shock andVibrio anguillavium-challenged stresses. Cold shock and WSSV treatment had no significant effects on the levels of HSP70 expression in all tissues examined. HSP70 induction was greatest after 2 h exposure to heat shock stress, which was elevated after acute heat shock exposure of 10°C above ambient temperature. Supported by the National Key Fundamental Research Program “Study of the Major Diseases and Resistance Mechanism of Mariculture Organisms”, NSFC, No. 30140017, and the international cooperative program “Immunaqua” by the European Commission, No. ICA4-CT-2001-10023.     

不同冷却方式下高温花岗岩巴西劈裂及声发射特性试验研究

干热岩地热开发中的钻井、储层压裂及热交换等环节均涉及高温岩石冷却的问题,为揭示其中岩石损伤演化规律,基于巴西劈裂试验和声发射技术,研究了不同高温及冷却方式对花岗岩抗拉性质的影响。结果表明:①25~600℃下花岗岩抗拉强度随温度升高而下降,遇水冷却使抗拉强度进-步下降并使其开始大幅下降的温度阈值提前到200,500℃后抗拉强度对遇水冷却更敏感。②荷载达到峰值,声发射累计振铃计数突增,岩样内形成断裂区;受遇水冷却影响,岩样的振铃计数峰值和能量峰值有所下降,间接反映岩石内裂纹更发育,200~300℃时降幅均较大,300℃时和500℃后花岗岩对热处理方式较敏感。③花岗岩破裂面随温度升高由平整向粗糙曲折变化,由脆性向延性转变,遇水冷却促进岩石破裂并促使脆性向延性转变的温度区间提前。研究结果为地热开采中高温岩石的稳定性评价提供理论参考。      

Rheological and Structural Properties of Sea Cucumber Stichopus japonicus During Heat Treatment

Changes in tissue structure, rheological properties and water content of raw and heated sea cucumber meat were studied. Sea cucumber Stichopus japonicus was heated at 25℃, 70℃ and 100℃ water for 5 min. The structural changes were observed using a light microscope and the rheological parameters （rupture strength, adhesive strength and deformation） determined using a texture meter. Microscopic photograph revealed that the structural change of heated meat was greater than that of raw meat. The rupture strength, adhesive strength and deformation of raw meat were smaller than those of the heated meat. Meanwhile, rheological parameters showed positive correlation with heating temperature. These changes are mainly caused by thermal denaturation and gelatinization of collagen during heating. These changes were also evidenced in observations using a light microscope and differential scanning calorimetry.     

Pyrolysis of Huadian oil shale by electrical heating on different heating rates

The electrical heating experiments on oil shale sample from Huadian of Jilin were carried out by the pyrolysis method at three different heating rate 2℃/ min,5℃/ min and 10℃/ min in the temperature range of 30℃-750℃ . Heating rate 2℃/ min is considered low,while intermediate one covers the range 5℃/ min and high heating rate is 10℃/ min. The controlling parameters studied were the final pyrolysis temperature and the influence of the heating rate as well as type. The heating rate has an important effect on the pyrolysis of oil shale and the amount of residual carbon obtained therefore. It is found that increasing the heating rate and pyrolysis temperature also increases the production of oil and the total weight loss. Higher heating rates resulted in higher rates of accumulation. The rate of oil and water collection passed through the maximum of different heating rates at different pyrolysis temperatures. Heating rate affected density,oil conversion and oil yield.