双棘黄姑鱼卵巢发育的周年变化

观察了双棘黄姑鱼卵巢发育的周年变化,结果表明,双棘黄姑鱼雌鱼性腺成熟系数(GSI)一年有两个峰值,分别出现在5月份(9.38%±0.32%)和10月份(7.56%±0.43%);HSI与GSI密切相关[R=0.76];卵巢发育水温在19～29℃范围,繁殖水温为24～28℃,在广东省饶平海区,双棘黄姑鱼繁殖时间在每年的5月和10月,属于春、秋二季产卵类型的鱼类。     

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。     

DNA甲基化修饰对金钱鱼卵巢Cyp17a1表达水平的影响

【目的】分析DNA甲基化修饰对金钱鱼（Scatophagus argus）卵巢Cyp17a1表达的影响,进一步认识卵巢发育成熟相关基因表达的调控机制。【方法】分别取卵巢发育III、IV期的金钱鱼,以及投喂添加0（对照）、50、100μg/g雌二醇饲料30 d的2龄雌鱼,用MethPrimer软件分析其Cyp17a1基因CpG二核苷酸位点分布特征,并预测CpG岛;采用亚硫酸氢盐测序法检测不同发育时期卵巢以及投喂雌二醇个体卵巢中的Cyp17a1的DNA甲基化修饰水平,并用实时荧光定量PCR分析Cyp17a1基因表达水平。【结果】金钱鱼Cyp17a1翻译起始位点2 000 bp后无CpG岛,取第1外显子含有5个CpG位点（翻译起始位点后106、116、129、148和203 bp处）的区域用于DNA甲基化水平检测。金钱鱼III期卵巢Cyp17a1外显子1的DNA甲基化修饰水平显著高于IV期卵巢（P <0.05）,与其mRNA表达水平呈负相关。116、129和203 bp处DNA甲基化存在发育时期差异,但106和148 bp处差异不显著（P> 0.05）。投喂雌二醇后,金钱鱼卵巢C...     

温度和盐度对军曹鱼幼鱼生长与抗氧化酶活性的影响

在实验室条件下,采用两因素交叉分组的方法,初步研究了环境因子(温度和盐度)对军曹鱼幼鱼肌肉抗氧化酶比活力的影响。温度设3个水平(26、29、32),盐度设4个水平(11、19、27、35),两个因素按其各自水平共产生12种组合。军曹鱼幼鱼在玻璃钢水槽内养殖20d后取样测定超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GPX)的比活力。结果表明:盐度因子诱导了军曹鱼的抗氧化应激反应,抗氧化酶活力随盐度降低而升高,SOD、CAT和GPX比活力分别从35时的17.74±3.38、19.44±2.66和13.70±2.99U/mg上升到11时的30.14±1.76、35.63±10.59和17.44±4.32U/mg;温度、盐度及因素间的交互作用对SOD和CAT比活力有显著影响(P<0.05);温度对GPX比活力无显著影响(P>0.05),而盐度及因素间的交互作用对GPX比活力影响显著(P<0.05)。     

不同培养液添加物对尼罗罗非鱼精原干细胞生长增殖的影响

【目的】探讨不同血清和细胞因子等培养液添加物对尼罗罗非鱼(Oreochromisniloticus)精原干细胞(Spermatogonial stem cells,SSCs)生长增殖的影响。【方法】无菌取发育第Ⅲ期的尼罗罗非鱼精巢组织,用酶消化法结合差速贴壁法获得尼罗罗非鱼SSCs,在有支持细胞饲养层条件下,L-15培养液中分别添加体积分数2%、5%、10%新生牛血清(New bovine serum,NBS),体积分数1%、2%、3%罗非鱼血清以及5、10、15 ng/mL白血病抑制因子(Leukemia inhibitor factor,LIF),比较不同添加物对尼罗罗非鱼SSCs生长增殖的影响。【结果】在支持细胞饲养层上,SSCs分裂指数明显高于无饲养层的SSCs,支持细胞饲养层明显促进精原干细胞分裂增殖(P 0.01);与2%、10%NBS组相比,添加体积分数5%NBS可显著促进精原干细胞增殖(P 0.01);添加体积分数1%、2%、3%尼罗罗非鱼血清对SSCs无显著的促增殖作用(P 0.05);添加5、10 ng/mL LIF可促进SSCs增殖,添加10 ng/mL LIF效果更佳,添加15 ng/mL LIF则抑制SSCs增殖。【结论】使用尼罗罗非鱼精巢支持细胞作饲养层,用添加体积分数5%NBS及10 ng/mL LIF的L-15培养液培养,有助于促进尼罗罗非鱼精原干细胞生长增殖。     

不同条件延缓未受精卵激活对鱼类细胞电融合的影响

泥鳅未受精卵先在合成卵巢液、郝氏液中分别浸泡15、20、30min,然后再受精孵化,统计发育情况,结果表明,合成卵巢液可有效延缓未受精卵遇水后被激活的时间,郝氏液对延缓未受精卵被激活也有一定作用,但效果比合成卵巢液差。用合成卵巢液配制不同浓度胰酶处理受体卵进行细胞电融合实验,胰酶质量浓度为5 mg/mL时,融合卵发育至囊胚期的比例比对照组(郝氏液)高2%,统计分析表明该差异是显著的(P<0.05);但发育至原肠期及孵出期的比例则与对照组无显著差异。胰酶质量浓度为2和1 mg/mL时,融合卵发育到各阶段的比例均低于对照组,统计分析表明两者的差异是显著的(P<0.05)。研究表明,溶液种类、胰酶浓度等对电融合卵的发育有明显的影响,但延缓受体卵的激活时间并不能提高电融合卵的出苗率。用郝氏液配制的2 mg/mL的胰酶溶液处理受体卵,可获得较高比例的融合鱼。     

太平洋褶柔鱼秋生群资源补充量预报模型研究

根据日本对太平洋褶柔鱼秋生群体的资源评估报告,以及产卵场海表温度(tSST)、叶绿素a质量浓度(ρChl-a),计算分析太平洋褶柔鱼在产卵期产卵场各月最适表温范围占总面积的比例(PS)、表征产卵场环境的tSST、ρChl-a等多种环境因子与单位捕捞努力量的渔获量(CPUE)的相关性,建立多种基于主要环境因子的资源补充量预报模型。结果表明:太平洋褶柔鱼资源补充量与各月的tSST的相关系数最大值海域分别为10月份的Point1(33.5°N,129.5°E)、11月份的Point2(31°N,127°E)和12月份的Point3(33.5°N,125°E);与各月的ρChl-a浓度的相关系数最大值海域出现在11月份的Point4(34°N,129.5°E)和12月份的Point5(35°N,130°E)。基于Point1的tSST、Point4的ρChl-a、PS等3个因子作为输入层构建的3-2-1的BP网络结构,2011-2012年的平均预报精度达到最高,为91.5%,该模型可用于太平洋褶柔鱼资源补充量的预测。     

花染色体组型分析及细胞核DNA含量的测定

采用PHA、秋水仙碱腹腔或背部肌肉注射,活体培养法,取花肾细胞制成悬浮液,用空气干燥法制片。经Giemsa染色,镜检细胞分裂相,统计结果为2n=50,核型公式为:2n=20m+12sm+10st+8t,NF=82。同时,取花肌肉和肝脏为材料,以鸡血细胞为DNA标准(2.30pg/2c),使用EPICS-XL型流式细胞仪测定了花细胞DNA含量,分别为2.219±0.055pg/2c和2.214±0.083pg/2c。     

LHRH-A和hCG对南方鲶血清促性腺激素水平的影响

运用放射免疫分析法测定了促黄体释放激素类似物 (LHRH A)和人绒毛膜促性腺激素(hCG)对南方鲶血清促性腺激素 (GTH)水平的影响 ,结果表明 :LHRH A和hCG都能提高南方鲶一龄幼鱼血清GTH水平 ,hCG与LHRH A结合注射后血清GTH水平显著高于单独注射hCG或LHRH A后的血清GTH水平。LHRH A和hCG结合注射可诱导性成熟南方鲶排卵 ,同时排卵过程中伴随着血清GTH水平的急剧上升 ,在排卵后迅速下降 ;注射同样剂量的药物而未排卵的血清GTH水平虽有升高 ,但升高的幅度不及排卵鱼。这种血清GTH水平的急剧增高对诱导南方鲶卵母细胞最后成熟和排卵具有重要作用。     

不同天然饵料对凡纳滨对虾亲虾性腺发育的影响

分别以双齿围沙蚕(Perinereis aibuhitensis)、杜氏枪乌贼(Loligo duvaucelii)、近江牡蛎(Ostrea riuularis)及三者的混合饵料(1∶1∶1)培育凡纳滨对虾(Litopenaeus vannamei)亲虾至性腺发育成熟,通过分析亲虾性腺发育时间、蜕壳间期、交配率、产卵量、变态率,确定培育亲虾的优质饵料。结果表明:投喂双齿围沙蚕或混合饵料的组与投喂杜氏枪乌贼、近江牡蛎组在产卵量、卵巢发育时间、蜕壳间期、交配率、孵化率、变态率方面具有显著差异(P<0.01)。投喂双齿围沙蚕或混合饵料的亲虾平均产卵量可达21.2×104粒/尾,卵巢平均发育时间为101 h,平均蜕壳间期为218 h,平均交配率可达80%,平均孵化率可达62.99%,平均变态率可达82.44%。表明在凡纳滨对虾亲虾培育的过程中,沙蚕单独或与其他天然饵料混合投喂,较乌贼、牡蛎单独投喂对亲虾性腺发育具有更好的促进效果。     

Seasonal variation of biochemical components in clam (<Emphasis Type="Italic">Saxidomus purpuratus</Emphasis> Sowerby 1852) in relation to its reproductive cycle and the environmental condition of Sanggou Bay,China

Seasonal variation of biochemical components in clam (Saxidomus purpuratus Sowerby 1852) was investigated from March 2012 to February 2013 in relation to environmental condition of Sanggou Bay and the reproductive cycle of clam. According to the histological analysis, the reproductive cycle of S. purpuratus includes two distinctive phases: a total spent and inactive stage from November to January, and a gametogenesis stage, including ripeness and spawning, during the rest of the year. Gametes were generated at a low temperature (2.1°C) in February. Spawning took place once a year from June to October. The massive spawning occurred in August when the highest water temperature and chlorophyll a level could be observed. The key biochemical components (glycogen, protein and lipid) in five tissues (gonad, foot, mantle, siphon and adductor muscle) were analyzed. The glycogen content was high before gametogenesis, and decreased significantly during the gonad development in the gonad, mantle and foot of both females and males, suggesting that glycogen was an important energy source for gonad development. The protein and lipid contents increased in the ovary during the gonad development, demonstrating that they are the major organic components of oocytes. The lipid and protein contents decreased in the testis, implying that they can provide energy and material for spermatogenesis. The results also showed that protein stored in the mantle and foot could support the reproduction after the glycogen was depleted.     

Ingestion of Domoic Acid and Its Impact on King Scallop（Pecten maximus,Linnaeus 1758）

A simple method for spiking formulated feed with domoic acid （DA） was developed in this study. DA feed was prepared by mixing 0.15 mL 100 μg mL-1 DA with 0.1 g formulated feed, and drying the mixture at room temperature for 2 h. The prepared DA feed contained 0.19 pg DA per particle. Of the added DA, 46.72% was retained in the feed. Relatively high DA retention （about 50%） was recorded after DA feed was soaked in water for 2h. Exposure to DA feed for 7d did not cause the increase of tissue DA level of adult king scallop （Pecten maximus） significantly in 60 d. The increase of their gonad index after DA exposure was not significantly different from the control. No significant change in DA level was found in spermary, ovary or fertilized eggs after DA exposure. These results indicated that DA excretion may be more efficient than DA accumulation under the current experimental conditions, and the mechanism of domoic acid incorporation in P. maximus may involve intracellular biotransformation.     

Hydrobiological variables as a regulatory factor on the abundance of heterotrophic flagellates in an urban pond

The seasonal abundance of flagellates has been monitored over a period of 1 year from December2013 to November 2014(divided into 4 conjugative seasons namely winter, spring, summer, and autumn) in an experimental pond located in Rajshahi City Corporation area, Bangladesh. To our knowledge, this study is the first to shed light on the occurrence and possible interrelationships among heterotrophic flagellates(HF),bacteria and zooplankton in Bangladesh and the result obtained by this study will be beneficial for similar water ecosystem all over the world. Standard methods were used to determine the prescribed hydrological parameters and zooplankton cell density. Maximum HF abundance(14 346.00 cells/mL) was found in the spring and the minimum(5 215.00 cells/mL) occurred in the summer. Inverse to HF, significantly(P0.05)higher zooplankton abundance was found during the winter(782.00±47.62 cells/mL) and the lowest value was found in the autumn(448.00±39.15 cells/mL). Whereas similar to the HF, total bacterial abundance was significantly higher during the spring((2.25±1.05)×10~5 cells/mL) and lower in the summer((0.79±0.06)×105 cells/mL). Multivariate analyses(ANOSIM and MDS) have shown significant seasonal differences for cell numbers where MDS ordination plot and cluster analysis based on similarity in the genera abundance of HF revealed overlapping condition between winter and spring. Canonical correspondence analysis(CCA) also showed a distinct separation among the genera based on the prevailing hydrological situation and indicated that temperature, pH, BOD_5, and NO_3~- were the most important environmental variables in determining the observed variation in HF community structure. Among the biological factors, zooplankton showed negative but total bacteria were positively correlated with HF abundance.     

Picoplankton distribution in different water masses of the East China Sea in autumn and winter

Picoplankton distribution was investigated in different water masses of the East China Sea in November,2006 and February,2007.The autumn and winter cruises crossed three major water masses:the coastal water mass(CWM),the mixed water mass(MWM),which forms on the continental shelf,and the Kuroshio water mass(KWM).Picoplankton composition was resolved into four main groups by flow cytometry,namely Synechococcus,Prochlorococcus,picoeukaryotes,and heterotrophic bacteria.The average abundances of Synechococcus,picoeukaryotes,and heterotrophic bacteria were(0.63±10.88)×103,(1.61±1.16)×103,(3.39±1.27)×105 cells/mL in autumn and(6.45±8.60)×103,(3.23±2.63)×103,(3.76±1.37)×105 cells/mL in winter,respectively.Prochlorococcus was not found in the CWM and seldom observed in surface samples in either season.However,Prochlorococcus was observed in the MWM and KWM(approximately 10 3 cells/mL) in both autumn and winter.Synechococcus distribution varied considerably among water masses,with the highest levels in KWM and lowest levels in CWM.The depth-averaged integrated abundance of Synechococcus was approximately 5-fold higher in KWM than in CWM,which may be due primarily to water temperature.In the MWM,Synechococcus was resolved as two subgroups;the presence of both subgroups was more common in autumn.Picoeukaryote abundance varied less among water masses than Synechococcus,and heterotrophic bacteria depth-averaged integrated abundance exhibited the smallest seasonal variations with respect to water mass.Correlation analysis showed that relationships between picoplankton abundances and environmental factors(temperature,nutrients,and chlorophyll a) differed among the three water masses,suggesting that the three water masses have different effects on picoplankton distribution(particularly Synechococcus).     

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.     

Retreived bacteria from Noctiluca miliaris (green) bloom of the northeastern Arabian Sea

In recent years, seasonal blooms of the dinoflagellate Noctiluca miliaris have appeared in the open-waters of the northern Arabian Sea (NAS). This study provides the first characterization of bacteria from a seasonal bloom of green Noctiluca of NAS (20°N?17°N and 64°E–70°E), during the spring-inter-monsoon cruise of Sagar Sampada 253, in March 2007. Bacterial growth as assessed by most-probable number (MPN) and plate counts, revealed ‘variable-physiotypes’ over a wide range of salinities (0%–25% w/v NaCl), pH levels (5–8.5), and organic nutrient strengths, in comparison to non-bloom waters. MPN indices of bacteria in surface waters of bloom stations *DWK and *PRB, corresponded to (3.08–4.41)×10³ cells/mL at 3.5% NaCl (w/v), and (2.82–9.49)×10² cells/mL at 25% (w/v) NaCl in tryptone-yeast extract broth (TYE). Plate counts were (1.12–4)×10⁶ CFU/mL at 0% (w/v) NaCl, (1.28–3.9)×10⁶ CFU/mL at 3.5% (w/v) NaCl, and (0.4–7)×10⁴ CFU/mL at 25% NaCl (w/v) on TYE. One-tenth-strength Zobell’s gave (0.6–3.74)×10⁵ CFU/mL at pH 5 to (3.58–7.5)×10⁵ CFU/mL at pH 8.5. These bacteria were identified to the genera Bacillus, Cellulomonas, Staphylococcus, Planococcus, Dietzia, Virgibacillus, Micrococcus, Sporosarcinae, Leucobacter, and Halomonas. The identity of three strains (GUFBSS253N2, GUFBSS253N30, and GUFBSS253N84) was confirmed through 16S rDNA sequence homology as Bacillus cohnii, Bacillus flexus, and Bacillus cereus. The ～2–3-fold higher plate counts of culturable bacteria from the open-waters of the NAS indicate that these bacteria could critically determine the biogeochemical dynamics of the bloom and its milieu. The role of these bacteria in sustaining/terminating the bloom is under evaluation.     

Effects of algal concentration and initial density on the population growth of Diaphanosoma celebensis Stingelin （Crustacea, Cladocera）

The effects of algal concentration and initial density on the population growth of the estuarine cladocera,Diaphanosoma celebensis Stingelin,were evaluated in an indoor experiment.A 2 × 4 layout that included two algal concentrations(Chlorella pyrenoidosa,1 × 106 and 3 × 106 cell/mL) and four inoculation densities(100,200,300 and 400 ind./L) were established.Diaphanosoma celebensis were reared in 150 mL flasks containing 50 mL of algal medium at 22°C,under salinity of 10 and a photoperiod of 12 h L:12 h D.T...