生物采油解堵剂中产蛋白酶菌株的筛选及培养条件优化

为研究生物酶采油解堵剂中产蛋白酶菌株的初、复筛选及培养条件优化,从大庆原油样品中筛选菌种,通过水解酪素的透明圈实验及福林酚测蛋白酶酶活的方法进行菌株的初、复筛选;以蛋白酶酶活为优化指标,采用单因素实验对筛选的产蛋白酶菌株的培养基及培养条件进行优化,优化最适培养基:可溶性淀粉为15g/L,蛋白胨为20g/L,酵母膏为20g/L,NaCl为1.0g/L,CaCl2为0.02g/L,Na2HPO4为0.2g/L,NaH2PO4为0.1g/L;在初始pH为6.0、接种量为5%(体积分数)、温度为31℃、摇床转速为160r/min的条件下,培养72h后,菌株的蛋白酶酶活为551.0U/mL,为复筛选菌株的蛋白酶酶活的22.92倍,即为菌株生长繁殖及代谢的最佳条件,能够获得更高的蛋白酶酶活,有利于后续实验的进行.结果表明:菌株产蛋白酶对原油作用效果为发酵液表面张力从作用前的56.2mN/m降低到作用后的30.5mN/m,表面张力显著降低,还有降解降黏原油等效果,具有一定的研究价值.     

东海盆地西湖凹陷不同构造带原油陆源生物标志化合物特征及其指示意义

利用地球化学分析手段,对西湖凹陷黄岩构造带、平北地区以及宁波构造带的16个轻质油样品进行了分析。原油中富含反映陆源高等植物输入的生物标志化合物,尤其是源于被子植物、裸子植物及蕨类植物的三环、四环二萜类及其芳构化产物,而反映水生低等植物输入的生物标志化合物含量较低。不同构造带间原油中这些化合物的分布特征具有明显差异,如异海松烷/nC₂₀、异海松烷/（异海松烷+16β（H）扁枝烷）、1,2,5-三甲基萘/1,3,6-三甲基萘、1,7-二甲基菲/（惹烯+1,7-二甲基菲）等。这些差异指示了宁波构造带与平北地区原油裸子植物输入较多,蕨类植物及被子植物输入较少,而黄岩构造带原油蕨类植物输入较多,被子植物与裸子植物较少的特点。研究中利用全油色谱-质谱将原油饱和烃与芳烃组分建立联系,提出（异海松烷+1,7-二甲基菲）/（惹烯+16β（H）扁枝烷）作为反映西湖凹陷轻质原油生源构成的指标,而该参数与全油碳同位素值的正相关关系证明了裸子植物树脂对西湖凹陷轻质油的重要成烃贡献。      

EFFECTS OF FISH OIL,DHA OIL AND LECITIHIN IN MICROPARTICULATE DIETS ON STRESS TOLERANCE OF LARVAL GILTHEAD SEABREAM（SPARUS AURATA）

The effects of natural fish oil,DHA oil and soybean lecithin in microparticulate diets on stress tolerance of larval gilthead seabream(Sparus aurata)were investigated after 15 days feeding trials.The tolerance of larval gilthead seabream to various stress factors such as exposure to air(lack of dissolved oxygen),changes in water temperature(low)and salinity(high) were determined.This study showed that microparticulate diet with natural fish oil and soybean lecithin was the most effective for in-creasing the tolerance of larval gilthead seabream to various stresses,and that microparticulate diet with natural fish oil and palmitic acid(16:0)was more effective than microparticulate diet with DHA oil and soybean lecithin.     

EFFECTS OF FISH OIL, DHA OIL AND LECITHIN IN MICROPARTICULATE DIETS ON STRESS TOLERANCE OF LARVAL GILTHEAD SEABREAM (SPARUS AURATA)

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Characteristics and sources of ore-forming fluids of South Narimalahei copper polymetallic deposit in East Kunlun,Qinghai

The South Narimalahei area is located on the north side of the Middle Kunlun fault in the eastern section of the East Kunlun composite orogenic belt. The ore body is veined and controlled by structures and se-condary fissures, which occurs in the structural alteration fracture zone in the Late Triassic granodiorite. In this deposit, copper mineralization is closely related to silicification and sericification. The formation process of the deposit includes hydrothermal mineralization and supergene oxidation. In this paper, the fluid inclusion minera-logy, microscopic temperature measurement and stable isotope studies have been carried out for ore of the main mineralization stage. The results show that the primary gas-liquid two-phase inclusions and a small amount of single-liquid inclusions are mainly developed in the quartz in the main mineralization stage. The results of microscopic temperature measurement show that the ore-forming fluid which has low temperature(151.7℃--205.8℃), low salinity(2.06 wt%--4.94 wt%NaCl), low density(0.86--0.92 g/cm~3) and shallow formation(1.5--3.0 km) is a hydrothermal solution of NaCl-H_2O system. Hydrogen and oxygen isotope results show that the ore-forming fluids mainly come from atmospheric precipitation, with a small amount of magmatic fluids participating. It is preliminarily determined that the South Narimalahei copper polymetallic deposit is a low-temperature hydrothermal vein deposit.     

Optimization of culturing condition and medium composition for the production of alginate lyase by a marine <Emphasis Type="Italic">Vibrio</Emphasis> sp. YKW-34

Carbohydrases secreted by marine Vibrio sp. YKW-34 with strong Laminaria cell wall degrading ability were screened, and among them alginate lyase was found to be dominant. The effects of medium composition and culturing condition on the production of alginate lyase by marine Vibrio sp. YKW-34 in flask were investigated in this study. In the culture medium of marine broth, no alginate lyase was produced. The activity of the alginate lyase, after being induced, reached 5 UmL^-1. The best inoculum volume and inoculum age were 10% and 12h, respectively. The optimal temperature for alginate lyase production was 25℃. The fermentation medium was composed of 0.5% of Laminaria powder and 0.2% of KNO3 with an initial acidity of pH 8.0. Alginate could induce alginate lyase production but not as efficiently as Laminaria powder did. The addition of fucoidan, cellulose and glucose had negative effect on the alginate lyase production. Other kinds of nitrogen sources, such as yeast extract, beef extract and peptone, had positive effect on the growth of the microorganism and negative effect on alginate lyase production. In addition, the time course of alginate lyase production under the optimized condition was described. The optimal harvest time was 48 h.     

Geochemistry and genesis on saturated hydrocarbon of crude oils in Gas Hure Oilfield in Qaidam Basin

Gas Hure Oilfield is the biggest one in the Qaidam Basin.Through intensive sampling and GC-MS analysis,the authors analyzed the geochemical characteristics of the crude oils in the Gas Hure Oilfield.The results show that the crude oils have even-odd carbon predominance for n-alkanes.In shallow reservoir,part of crude oils suffered biogenic degradation.As for normal oils,they have low Pr/Ph ratios with abundant gammacerane and C35 homohopane,which indicate that the crude oils are formed in strong reducing salt water environment.Compared deep(E13) reservoir with shallow(N1-N12) reservoir,the ancient salinity of sedimentary environment of N1--N12 crude oils is slightly higher than that of E13 crude oils;the C-21/C+22 ratios of crude oils are 1.02-1.71 with a higher abundance of C27 sterane,which shows that their mainly origin matrix are bacteria and algae;the C29 sterane 20S/(20S+20R)is generally low,the ratio in deep E13 reservoir distributes between 0.31 and 0.40,but in shallow reservoir is relatively low with 0.29-0.36.At the same time,C29 sterane αββ/(αββ+ααα)and Ts/Tm both are extremely lower.All the above show that the crude oils in Gas Hure are mainly low mature oils,although the maturity of deep reservoir crude oils is slightly higher than shallow one.     

文昌鱼对几种环境因子及污染物的耐受极限初步研究

<正>文昌鱼(Branchiostoma belcheri)隶属脊索动物门头索动物亚门,是全世界现存动物中与脊椎动物关系最近的一类非脊椎动物[1],被称为生物进化的“活化石”。它营养丰富,干品蛋白质含量约达70%左右,还含有较高的磷和碘,脂肪含量低,具有较高的     

减氧空气与轻质原油低温氧化反应特征

塔河北区块轻质原油与减氧空气在温度和压力作用下,探究其组分变化情况和耗氧量规律。在室内使用高温高压反应釜开展原油静态氧化实验,较为系统地研究不同含氧量及不同注气量的减氧空气对原油的氧化作用;并利用色谱分析对烃类分析,收集反应气体进行气体分析,气体组分和原油组分都有不同程度的变化。研究结果表明：(1)原油样本与减氧空气发生低温氧化反应,重质组分C₂₁以上的组分均下降,低于C₁₃的轻质组分含量均上升;(2)原油氧化效果随着含氧量上升而增强,反应过程组分对应“拐点”靠前;(3)含氧量一定时,改变油气比,反应对应的“拐点”不变;含氧量为下降趋势后趋于平缓;(4)低温氧化过程生成的CO₂含量为上升趋势后趋于平缓;(5)在油藏温度(160℃)下,原油主要涉及2个反应热裂解和加氧反应。热裂解反应与含氧量无关,和温度有关,主要反应温度是160～200℃。高碳组分沥青质等重质组分会分解成饱和烃。加氧反应主要是芳香烃,胶质加氧反应也会生成沥青质。当2种反应同时存在时,沥青质参与热裂解的含量大于加氧反应芳香烃等生成的沥青质量,会导致沥青质含量...     

温度、盐度、pH和体质量对尼罗罗非鱼幼鱼窒息点的影响

利用实验生态学的方法研究不同温度(15、20、25、30和35℃)、盐度(0、5、10、15和20)、pH(5、6、7、8和9)和体质量(1.93±0.08、3.43±0.12、7.95±0.34、11.74±0.41和15.84±0.61 g)对尼罗罗非鱼幼鱼窒息点的影响,并建立温度、盐度、pH、体质量与罗非鱼窒息点间的回归方程。结果表明:温度t、盐度S、pH和体质量m对尼罗罗非鱼幼鱼窒息点有显著影响(P<0.05)。当温度、盐度、pH和体质量分别为25.28℃、3.6、8.06、6.3 g时,窒息点(DAP)最低,回归方程为DAP=0.000 9 t 2-0.045 5 t+0.726 6(R2=0.952 1),DAP=-0.000 1 S 3+0.004 2 S 2-0.023 9 S+0.111 9(R2=0.922 8),DAP=0.009 3 pH2-0.149 9 pH+0.7174(R2=0.976 8);DAP=-0.000 3 m3+0.011 1 m2-0.104 3 m+0.362 8(R2=0.948 4)。在溶氧不足情况下,温度25.28℃、盐度3.6、pH 8.06的环境条件有助于提高幼鱼耐低氧能力。     

盐度突变对青蛤(Cyclinasinensis)鳃Na~+/K~+-ATPase活性的影响

研究了盐度23下青蛤(Cyclina sinensis)鳃、斧足、外套膜和内脏团中Na+/K+-ATPase的活性差异,并以活性高的鳃组织为对象,对青蛤实施了低盐(23→16)和高盐(23→36)突变的胁迫,监测了鳃组织中Na+/K+-ATPase活性的动态变化。结果表明,盐度23下青蛤鳃组织中Na+/K+-ATPase的活性显著高于其他组织;在盐度突降胁迫下,鳃组织中Na+/K+-ATPase的活性于前16 d逐渐升高,第17d起显著升高(p<0.05),第18 d达到峰值,于第20 d回落,但仍高于起始水平;在盐度骤升胁迫下,鳃组织中Na+/K+-ATPase的活性于前13d逐渐降低,第14 d起显著降低(p<0.05),于第15 d达到最低值,继而回升至平稳水平,但仍低于起始水平。青蛤不同盐度胁迫下其鳃组织中Na+/K+-ATPase活性的响应方式不同,低盐时活性增加,而高盐时活性下降;此外,青蛤在盐度胁迫下对自身渗透压的调节则需较长时间。     

Nutrient-enhanced n-alkanes biodegradation and succession of bacterial communities

Bioremediation, is an effective and environment-friendly method of cleaning up crude oil pollution after an oil spill. However, the in situ bioremediation of crude oil is usually inhibited by deficiency of inorganic nutrients. To understand the effects of nutrient addition on the bioremediation of crude oil and the succession of bacterial communities during process of bioremediation, microcosms containing oilcontaminated sediments were constructed and biodegradation of crude oil was assessed based on the depletion of different ingredients. We used two culture-independent methods, denaturing gradient gel electrophoresis and a 16 S rRNA gene based clone library, to analyze the succession of bacterial communities. The results suggested n-alkanes were degraded after 30 days and that nutrient amendments significantly improved the efficiency of their biodegradation. Moreover, oil contamination and nutrient amendments could dramatically change bacterial community structures. Lower diversity was detected after being contaminated with oil. For instance, bacterial clones affiliated with the phylum Armatimonadetes, Firmicutes, Gemmatimonadetes, and Planctomycetes and the class Deltaproteobacteria and Epsilonproteobacteria could not be identified after 30 days of incubation with crude oil. However, "professional hydrocarbonocastic bacteria" became abundant in samples treated with oil during the bioremediation period, while these clones were almost completely absent from the control plots. Interestingly, bioinformatics analysis showed that even when dramatic differences in oil biodegradation efficiency were observed, bacterial communities in the plots with nutrient amendments were not significantly different from those in plots treated with oil alone. These findings indicated that nutrient amendments could stimulate the process of biodegradation but had less impact on bacterial communities. Overall, nutrient amendments might be able to stimulate the growth of n-alkane degrading bacteria.     

Allelopathic effects of macroalga <Emphasis Type="Italic">Corallina pilulifera</Emphasis> on the red-tide forming alga <Emphasis Type="Italic">Heterosigma akashiwo</Emphasis> under laboratory conditions

Over the past few years, harmful algal blooms (HABs), such as red tides, have been frequently observed in coastal zones worldwide. The natural allelopathic interactions among macroalgae and red tide microalgae can alter the structure and succession of aquatic ecosystems. We investigated the influence of four environmental factors (temperature, salinity, light, and pH) on the allelopathic effects of the macroalgae Corallina pilulifera on red-tide forming Heterosigma akashiwo under laboratory conditions. Each of the factors had four levels: temperature (15, 20, 25, and 30°C), salinity (10, 20, 30, and 40), light (20, 100, 200 and 400 μmol/(m²?s)), and pH (5.5, 7, 8.5, and 10. Two-factor experiments were designed for each two environmental factors, with six combination treatments (temperature-salinity, temperature-light, temperature-pH, salinity-light, salinity-pH, and light-pH). Results showed that the allelopathic effect was significantly influenced by temperature, salinity, light, and pH. As single factors, the low temperature (15°C), low salinity (10), high-intensity light (400 μmol/(m²?s)), and high pH (10) treatments substantially enhanced the allelopathic effect. The strongest allelopathic effect of C. pilulifera on H. akashiwo was observed under the following treatments: 15°C and salinity of 40, 25°C and pH 10, 25°C with medium- to high-intensity light at 200–400 μmol/(m 2 ?s), 400 μmol/(m²?s) and salinity of 10, 400 μmol/(m²?s) and pH 10, and pH 10 with a salinity of 40.     

饶阳凹陷留西—留北地区新近系地层水特征及其与油气分布的关系

以冀中坳陷饶阳凹陷新近系油气分布最富集的留西—留北地区为研究对象,通过对该区新近系地层水及其与油气分布关系的系统分析,明确了新近系地层水的水化学类型及矿化度分布特征,揭示了该区新近系地层水的地球化学特征异常与油气运移、油气分布的内在关系。结果表明:留西—留北地区新近系整体上发育低矿化度的NaHCO3型地层水,地层水矿化度具有由西向东、由北向南逐渐增加的趋势,同时水化学类型由HCO3--Cl--Na+型过渡为Cl--HCO3--Na+型;在油源和断裂条件良好的条件下,新近系地层水异常与油气分布具有良好的对应关系,新近系油气主要在地层水矿化度高于背景值、水化学类型异常及油源断裂发育的地区富集,同时新近系高矿化度Na2SO4型或MgCl2型地层水对油气由深向浅的运聚特征具有重要的指示作用。     

墨西哥湾扇贝稚贝盐度适应性的研究

在室内控制条件下,观察不同海水盐度对墨西哥湾扇贝(Argopecten irradians concentricusSay)稚贝存活及生长的影响,并将168 h 50%死亡的盐度作为适宜生存临界盐度,将增长率为最佳增长率30%时所对应的盐度作为适宜生长临界盐度。结果表明:在稚贝壳高4.38 mm±0.31 mm,水温25.4~27.8℃,pH值7.8~8.2下,墨西哥湾扇贝稚贝适宜生存盐度为23.6~35.7,最适生存盐度为26.0~31.3;适宜生长盐度为23.9~35.8,最适生长盐度为28.7~31.3。在盐度变化超出最适范围时,`墨西哥湾扇贝稚贝对低盐更为敏感,高盐比低盐对稚贝存活、生长、增重的影响小。当盐度为28.7时,墨西哥湾扇贝稚贝具有最高的相对存活率(100%)、壳高日增长率(0.116 2 mm.d-1)及体重日增长率(1.95 mg.d-1)。当盐度在最适范围之外升高或降低,稚贝的存活率、壳高及体重日增长率也随着降低。     

鄂尔多斯盆地姬塬油田延长组原油性质与来源分析

通过原油与油田水物理性质、原油与烃源岩族组成、饱和烃气相色谱以及萜烷与甾烷等生物标志化合物特征的多项地球化学指标对比,表明鄂尔多斯盆地姬塬油田延长组长8、长6、长4+5等油层组原油性质较为复杂,其成熟度、母源氧化-还原环境及母质类型等方面存在显著差异。烃源岩评价也显示长9、长8、长7、长6、长4+5段的暗色泥岩有机质质量和类型有所差别,但均具备生排烃能力。油源对比结果进一步证明:成藏过程存在多套油源混源供油的成藏效应,其中长8油层组原油主要来源于长7—长8段烃源岩;长6油层组原油主要来源于长7—长6段烃源岩;长4+5油层组原油存在长7、长6及长4+5段多源烃源岩的贡献。     

泌阳凹陷北部缓坡带核桃园组油气运聚成藏过程

为了深入了解油气分布规律，利用流体包裹体显微荧光光谱和显微测温以及一维盆地模拟的埋藏热历史，结合地震构造属性和沉积相分析，对泌阳凹陷北部缓坡带核桃园组油气运聚成藏过程进行了研究。结果显示北部缓坡带核桃园组第三段储层检测到橙黄色、黄绿色和蓝绿色3种颜色荧光油包裹体，反映了3幕不同成熟度的原油捕获。油包裹体的丰度和伴生盐水包裹体均一温度和埋藏史曲线表明，北部储层共存在两期三幕充注事件:第一期规模小，发生于断陷末期的相对低成熟度发橙黄色荧光油，第二期发生在中新世构造抬升期的相对高成熟度的黄绿色和蓝绿色荧光油。流体包裹体古压力模拟显示第二期油气充注时储层是超压状态，推测与当时的构造挤压有关。超压持续作用以及北部地层抬升造成与深凹带源岩增加的流体势差为第二期油气提供了强有力的动力；频繁的断层活动，特别是走滑断裂使得单一的砂体顺层输导转变为砂体顺层输导+断裂垂直输导的复合立体运移输导体系，使得原油以高效率运移到凹陷北部缓坡带。这对精细理解构造抬升控制油气大规模运聚具有重要的意义。      

Distribution Patterns of Synechococcus Indicated Their Environmental Constraints and Related Geochemical Processes in the Yellow Sea

Synechococcus is a widely distributed photosynthetic pico-phytoplankton,which contributes mainly to carbon fixation and maintains the stability of the marine ecosystem.To investigate its distribution patterns in the Yellow Sea,seawater samples were collected during September 2018.Results of flow cytometry analysis showed that the Synechococcus abundance ranged from 6.36×10² to 4.51×10⁴ cells mL^?1,which correlated with salinity(P<0.01)and temperature(P<0.05).At deeper off-shore sites,Synecho-coccus showed high abundance at the subsurface thermo-halocline,which was in accordance with chlorophyll a(Chl a)content along the vertical column.Based on the high-throughput sequencing data of rpoC1(partial gene encoding RNA polymerase),two Synechococcus subclusters,S5.1 and S5.2,were found to coexist in the studied area.Several clades of S5.1,including Clades I,II,and III,were the dominant components,accounting for 6.63%,26.11%,and 45.5%of the total genus,respectively.Redundancy analysis(RDA)showed that nitrite was the main environmental factor that explained the genus composition among samples.Fur-thermore,co-occurrence network revealed that the main phyla that coexisted with Synechococcus were Proteobacteria,Bacteroidetes,Actinobacteria,Planctomycetes,and Verrucomicrobia,which were involved in the carbon(C),nitrogen(N),sulfur(S),and manga-nese(Mn)cycles.Overall,Synechococcus exhibited biogeographic distribution correlated with temperature-salinity and nitrite in the Yellow Sea,and their geochemical function showed diverse but should be further verified in the future.     

Removal of K^＋, Na^＋, Ca^2＋, and M^g2＋ from saline-alkaline water using the microalga Scenedesmus obliquus

The capability ofScenedesmus obliquus to remove cations （K^＋, Na^＋, Ca^2＋, Mg^2＋） from saline- alkaline water was investigated at different salinities （0, 5, 10, 15, 20, 25） and carbonate alkalinities （0, 5, 10, 15, 20, 25, 30, 35 mmol/L）. K^＋, Na^＋, Ca^2＋, and Mg^2＋ in saline-alkaline water were efficiently removed by S. obliquus. The maximum removal of the cations （29.37 mg for K^＋, 185.85 mg for Na^＋, 23.07 mg for Ca^2＋, 66.14 mg for Mg^2＋） occurred at salinity 25. The maximum removal of K^＋ （2.28 mg）, Na＋ （6.62 mg）, Ca^2＋ （1.01 mg）, and Mg2＋ （0.62 mg） occurred at carbonate alkalinities of 25 mmol/L for K＊, 35 mmol/L for Na＋, 20 mmol/L for Ca2＋, and 25 mmol/L for Mf＋, respectively. Under a salinity stress, the concentration of Na＇ in S. obliquus increased significantly, while that of K~ decreased significantly. The concentrations of Ca^2＋ and Mg2＋ decreased as well. The ratios of K＋/Na~, Ca2＋/Na^＋, and Mg^2＋/Na^＋ were significantly lower in all salinity treatments than those of the control. Under alkaline stress, the concentrations of Nan and K＋ in S. obliquus decreased significantly and the ratios of K^＋/Na^＋, Ca2＋/Na^＋, and Mg^2＋/Na^＋ were significantly higher in all treatments than in the control. Moreover, the concentrations of Ca2＋ and Mg2＋ in S. obliquus at alkalinities of 5-10 mmol/L were significantly higher than those of the other treatments. The removal of Na＋ by S. obliquus mainly occurs through biosorption, and Mg^2＋ and Ca^2 ＋ were removed through both biosorption and bioaccumulation.     

华贵栉孔扇贝稚贝盐度适应性的研究

通过室内实验研究了水温28.5～29.4℃、pH值7.6～8.1条件下,华贵栉孔扇贝Chlamys nobilis(Reeve)稚贝(壳高9.77±0.19mm)对不同盐度的适应性。结果表明:华贵栉孔扇贝稚贝适宜生存盐度为24.27~37.18,最适生存盐度为26.91~32.13,适宜生长盐度为24.56~37.09,最适生长盐度为26.91~29.52。在适宜生存盐度范围内,当盐度变化超出最适范围时,低盐端变化对相对成活率、壳高日增长率、体重日增长率影响比高盐端明显。