用于海底热液pH探测电极自校正的控制技术研究

在海底热液pH值的探测过程中,探测电极的特性会随时间的推移而发生变化。为了实现长期的原位探测,需要一套流体控制系统对探测电极进行自校正与自维护。针对所设计的流体控制系统,遵循低功耗的设计原则,研制了其主控系统,包括硬件设计和软件开发。为了使控制系统能应用于海底热液环境,对其进行了耐压封装设计,并进行了高压水实验,实验结果表明系统在40 MPa的高压下能正常工作。各种测试指标亦表明系统能自动实现pH探测电极自校正与自维护的功能。     

一种深海热液原位探测设备供能装置的设计

为了解决深海热液原位探测设备的能量供给问题,设计了一种实用的供能装置.该供能装置采用了一种定制的高能量密度电池,电池组的容量达31.5 Ah,最大输出电压为12 V.供能装置的耐压壳体采用了压力容器的设计原则进行设计,其最大承受压力达70MPa.放电实验表明该供能装置具有较稳定的放电电压;海试实验表明该供能装置能用于深...     

深海细菌Pseudomonas sp. IOFA1中甲醛歧化酶FM11的纯化及性质研究

从印度洋深海沉积物中分离出一株能够高效降解甲醛的假单胞菌(Pseudomonas sp.IOFA1),该菌株可在40 min内完全降解100 mg/cm~3的甲醛.通过阴离子交换柱层析对菌株破碎液中的粗酶液进行纯化,得到一条具有甲醛降解活性的单一蛋白条带.经MALDI-TOF/TOF质谱鉴定,该蛋白为甲醛歧化酶蛋白(FM11).序列分析表明FM11与其它物种已知醛脱氢酶的氨基酸序列最高相似性为65%;结构域分析表明FM11含有2个保守的锌结合位点和一个NAD结合位点.酶学性质分析结果显示,FM11的最适反应温度为40℃,在25~50℃范围内能够保留95%以上的酶活力,尤其是在高于70℃的高温环境下仍保有约50%的活力,表现出广泛的温度适应性.FM11具有非常广泛的最适反应pH值范围(pH值为5~9),且在pH值4和pH值10时仍分别保留50%和80%左右的酶活力,表现出广泛的pH适应性.在pH值为5~7的条件下孵育1 h后,FM11仍保有97%以上的酶活,且在pH值为7~10条件下孵育1 h后,仍保留90%以上的酶活,表现出极好的酸性和碱性稳定性.Ca~(2+)、Mg~(2+)在低浓度时(0.5 mmol/dm~3)对FM11有较明显的激活作用.这些研究为我们进一步开发Pseudomonas sp.IOFA1甲醛歧化酶在降解甲醛方面的用途奠定了基础.     

深海化能极端环境中劳盆拟刺铠虾(Munidopsis lauensis)的全长转录组测序分析

劳盆拟刺铠虾(Munidopsis lauensis)是一种十足目铠甲虾, 能够适应深海化能合成生态系统的极端环境, 但是其已知基因序列信息却很少。为此, 采用PacBio平台的SMRT测序技术对来自南海台西南冷泉劳盆拟刺铠虾的肝胰腺、鳃、肌肉和肠混合组织进行了三代全长转录组测序。通过聚类、校正、去冗余之后共获得28 811条高质量isoform, 平均长度为2 086 bp, N50长度为2 275 bp。使用Nr、SwissPort、KEGG、KOG数据库对转录本序列进行功能注释, 共有20 616 (71.56%)条isoform得到注释。进一步高级注释共获得21 848个蛋白编码序列, 共预测到537个转录因子、6 430个长链非编码RNA和10 060个SSRs位点。KEGG通路分析发现两条可能与劳盆拟刺铠虾适应深海化能极端生境相关的通路, 分别为过氧化物酶体通路和谷胱甘肽代谢通路。其中, 共有180条isoform被发现参与编码过氧化物酶体通路中的31个关键酶, 213条isoform参与谷胱甘肽代谢通路中19个关键酶的编码。基于全长转录组数据, 利用同源比对、功能结构域预测及系统发育树构建等方法对劳盆拟刺铠虾谷胱甘肽S-转移酶(GST)基因家族进行挖掘与鉴定, 共发现属于theta、delta、mu、kappa四种亚家族的20条GST候选序列。通过对劳盆拟刺铠虾全长转录组测序、功能注释和环境适应相关的重要基因及通路的分析, 不仅丰富了深海组学数据资源, 也为进一步揭示甲壳动物适应深海化能极端环境的分子机制奠定了基础。     

深海羽毛降解菌Bacillus sp. JM7极端碱耐受角蛋白酶的酶活性质研究

Bacillus sp. JM7, a strain isolated from the deep-sea of the South China Sea, was found to efficiently degrade 79.4% native chicken feather within 30 h. Scanning electron microscopy analysis showed that JM7 strain could gradually degrade feather by modifying the microstructure of feather keratin. A total of 25 protease genes were predicted from the draft genome of JM7 strain, among which a predicted subtilisin-like serine protease(designated as Ker02562) was further characterized for its keratinolytic activity. The recombinant Ker02562 functioned at a wide range of temperatures from 30℃ to 60℃, with an optimum at 40–50℃. Ker02562 was highly active at various pHs ranging from 5.0 to 13.0, with a maximum activity observed at pH 7.0–9.0. Remarkably, recombinant Ker02562 was stable in extreme alkaline environments(pH 10–13), which was much better than most other reported keratinases. Collectively, these favorable properties could make Bacillus sp. JM7 and Ker02562 attractive to be applied in the detergent formulation and feather bioconversion.     

深海热液区化能自养菌Caminibacter profundus氢酶对环境因子的响应特点

以深海热液区化能自养菌Caminibacter profundus为对象,采用扫描电镜和化学能谱分析,研究了菌株表面形态和矿物元素沉积;同时研究了菌株生长、甲基紫精(MV)还原的氢酶活性及膜结合的类型I NiFe氢酶基因(hynL)表达对盐度、pH和温度几种环境因子变化的响应特点。结果表明,菌株表面被Si、Ca、S和Fe等多种元素组成的矿物质层所包裹。不同条件下,菌株生长、MV还原的氢酶活性和hynL的表达趋势相一致,其最适条件为盐度30、pH5.5和55℃培养温度。研究结果表明,在环境改变时,C.profundus通过调控hynL的表达,以调整菌株的能量代谢,维系菌株的生长繁殖。     

一种新型电镀法制备的Ir/Ir(OH)x pH电极及其在加州纽波特港海域的应用研究

Resulting from the rising levels of atmospheric carbon, ocean acidification has become a global problem. It has significant impacts on the development, survival, growth and physiology of marine organisms. Therefore, a highprecision sensor is urgently needed to measure the pH of sea-water. Iridium wire with a diameter of 0.25 mm is used as the substrate, and an Ir/Ir(OH)_x pH electrode is prepared by a one-step electrochemical method in a Li OH solution at the room temperature. A scanning electron microscope(SEM) observation reveals that it is coated with nanoscale particles. In laboratory tests, the electrode exhibits a very promising pH response, with an ideal Nernst slope(56.14–59.52), fast response, good stability and long life-span in tested pH buffer solutions. For a sea trial,four pH electrodes and one Ag/Ag Cl reference electrode are integrated with a self-made chemical sensor, and a profile detection of nearly 70 m is implemented near Newport Harbor, California on August 3, 2015. The results reflect that the pH value measured by the sensor is very close to the data given by Sea-Bird 911 plus CTD, with a difference value ranging from 0.000 075 to 0.064 719. And the sensor shows a better data matching degree in 0–40m water depth. In addition, the high precision and accuracy of the sensor make it possible to use in the ocean observation field.     

涠11-4平台在极端环境条件下的人员工作安全性评估

本文以位于中国南海的涠１１－４平台为对象,进行了在强台风条件下的平台关键部位动力响应及环境参数的监测;并以测试数据为基础完成了动态建模与分析计算,给出了极端环境条件下生活模块的最大动态响应;评估了工作人员的适应能力及振动对人体健康与安全的影响。     

不同无机氮浓度下中肋骨条藻和三角褐指藻生长对生长环境pH的影响

海洋酸化是当前一个严重的环境问题,而海水pH的变化与微藻生长时对溶解无机碳的吸收有关。在室内条件下采用生理生态学方法,研究中肋骨条藻Skeletonema costatum和三角褐指藻Phaeodactylum tricornutum在不同无机氮水平下生长到最大生物量时培养液pH的变化。结果表明,培养液无机氮浓度从10μmol.L-1上升到200μmol.L-1过程中,2种微藻的叶绿素浓度和最终pH均呈上升趋势。培养液pH的增加量与无机氮的消耗量呈正相关关系,培养液pH的增加量与溶解无机碳的消耗量也呈正相关关系。     

Simultaneous vertical measurements of in situ pH and CO2 in the sea using spectrophotometric profilers

We have developed new systems capable of profiling to >1000 m for measuring in situ pH and fugacity of CO₂ (fCO₂) in the ocean using spectrophotometric analysis (pH and CO₂ profilers). The in situ pH is determined by detecting the color change of the pH indicator (m-cresol purple). It can withstand ambient pressure to 1000 m depth. The CO₂ profiler analyzed in situ fCO₂ by detecting the change of pH in an inner solution, equilibrated with the seawater through a gas permeable membrane. It can be operated to 2500 m depth. We used an amorphous fluoropolymer tubing form of AF-2400 for the gas permeable membrane due to its high gas permeability coefficients. The inner solution was a mixture of 2 μM bromocresol purple (BCP) and 5 μM sodium hydroxide. This system gave us a response time of 1 minute, which is twice as fast as previous systems. The precisions of pH and CO₂ profilers were within 0.002 and 2.5% respectively. We have used these profilers to study the North Pacific, obtaining good agreement with the difference between the data from profilers and a discrete bottle of 0.002 ± 0.005 pH (SE, n = 25) and −0.4 ± 3 μatm (SE, n = 31).     

Spectrophotometric determination of pH in seawater off Taiaroa Head,Otago, New Zealand: Full-spectrum modelling and prediction of pCO2 levels

Accurate measurement of seawater pH has long been sought by marine chemists (for example: [Dickson, A.G. 1993a. The measurement of sea water pH. Marine Chemistry, 44, 131–142, Dickson, A.G. 1993b. pH buffers for sea water media based on the total hydrogen ion concentration scale. Deep-Sea Research, 40, 107–118; Zhang, 1996; Tapp, M., Hunter, K.A., Currie, K. and Macaskill, B. 2000. Apparatus of continuous-flow underway spectrophotometric measurement of surface water pH. Marine Chemistry 72(2–4), 193–202; Friis, K., Koetzinger, A., Wallace, D.W.R. 2004. Spectrophotometric pH measurement in the ocean: Requirements, design and testing of an autonomous charge-coupled device detector system. Limnology and Oceanography: Methods 2, 126–136]. Recently, such attempts have taken on greater significance as anthropogenic carbon dioxide emissions may create rapidly changing oceanic pH. Spectrophotometric techniques have been accepted generally as the best for determination of seawater pH. Here we report a new technique using thymol blue as the indicator dye and fitting the entire spectrum from 400 to 900 nm rather than measuring the absorbance values at only two or three points in the spectrum. This full-spectrum modelling enables a reduction in signal to noise over other techniques. In the laboratory, we find with seawater samples a pH precision increase of five-fold “within” a sample and seven-fold “between” samples when comparing the full spectrum to the three-point method of analysis [Zhang, H., Byrne, R.H. 1996. Spectrophotometric pH measurements of surface seawater at in-situ conditions: absorbance and protonation behaviour of thymol blue. Marine Chemistry 52, 17–25].     

Lethality of increasing CO2 levels on deep-sea copepods in the western North Pacific

The first CO₂ exposure experiments on several species of pelagic copepods inhabiting surface and deep layers in the western North Pacific were conducted. Living organisms were collected from two layers between the surface and 1,500 m between latitudes of 11 and 44°N, and they were exposed aboard ship to various pCO₂ up to about 98,000 μatm. Mortality of copepods from both shallow and deep layers in subarctic to subtropical regions increased with increasing pCO₂ and exposure time. Deep-living copepods showed higher tolerance to pCO₂ than shallow-living copepods. Furthermore, deep-living copepods from subarctic and transitional regions had higher tolerances than the subtropical copepods. The higher tolerances of the deep-living copepods from subarctic and transitional regions may be due to the adaptation to the natural pCO₂ conditions in the subarctic ocean.     

Impurities in indicators used for spectrophotometric seawater pH measurements: Assessment and remedies

Spectrophotometric pH measurements have been widely used in oceanic CO₂−system research. However, impurities in indicator dyes from different manufacturers may cause uncertainty in measured pH values. In this work we report initial findings surrounding the potential significance of indicator sources (manufacturers) on pH measurements obtained with the sulfonephthalein indicator m-cresol purple. HPLC analyses were used for comparative assessments of impurities in m-cresol purple obtained from a range of manufacturers. Results indicate that m-cresol purple from different manufacturers, and perhaps from different batches of the same manufacturer, have different types and quantities of light-absorbing impurities. Impurities can contribute to pH offsets as large as 0.01 pH units. The pH offset caused by impurities decreases with decreasing sample pH. Indicators can be purified using preparative HPLC. Until purified indicators and refined indicator calibrations become available, we suggest that investigators should reserve some portion of each indicator batch for comparison with the properties of purified indicators.     

Physiological effects of hypercapnia in the deep-sea bivalve Acesta excavata (Fabricius, 1779) (Bivalvia; Limidae)

The option of storing CO₂ in subsea rock formations to mitigate future increases in atmospheric CO₂ may induce problems for animals in the deep sea. In the present study the deep-sea bivalve Acesta excavata was subjected to environmental hypercapnia (pHSW 6.35, PCO₂ = 33,000 μatm) corresponding to conditions reported from natural CO₂ seeps. Effects on acid–base status and metabolic rate were related to time of exposure and subsequent recovery. During exposure there was an uncompensated drop in both hemolymph and intracellular pH. Intracellular pH returned to control values, while extracellular pH remained significantly lower during recovery. Intracellular non-bicarbonate buffering capacity of the posterior adductor muscle of hypercapnic animals was significantly lower than control values, but this was not the case for the remaining tissues analyzed. Oxygen consumption initially dropped by 60%, but then increased during the final stages of exposure, which may suggest a higher tolerance to hypercapnia than expected for a deep-living species.     

Experiments on the ocean sequestration of fossil fuel CO2: pH measurements and hydrate formation

We have carried out a series of in situ experiments to investigate the formation of a CO₂ hydrate (CO₂:5.75 H₂O) for the purpose of evaluating scenarios for ocean fossil fuel CO₂ disposal with a solid hydrate as the sequestered form. The experiments were carried out with a remotely operated vehicle in Monterey Bay at a depth of 619 m. pH measurements made in close proximity to the hydrate–seawater interface showed a wide range of values, depending upon the method of injection and the surface area of the hydrate formed. Rapid injection of liquid CO₂ into an inverted beaker to form a flocculant mass of hydrate resulted in pH initially as low as 4.5 within a few centimeters of the interface, decaying slowly over 1–2 h towards normal seawater values as dense CO₂ rich brine drained from the hydrate mass. In a second experiment, slower injection of the liquid CO₂ to produce a simple two-layer system with a near planar interface of liquid CO₂ with a thin hydrate film yielded pH values indistinguishable from the in situ ocean background level of 7.6. Both field and laboratory results now show that the dissolution rate of a mass of CO₂ hydrate in seawater is slow but finite.     

Alterations in seawater pH and CO2 affect calcification and photosynthesis in the tropical coralline alga, Hydrolithon sp. (Rhodophyta)

Calcification in the marine environment is the basis for the accretion of carbonate in structures such as coral reefs, algal ridges and carbonate sands. Among the organisms responsible for such calcification are the Corallinaceae (Rhodophyta), recognised as major contributors to the process world-wide. Hydrolithon sp. is a coralline alga that often forms rhodoliths in the Western Indian Ocean. In Zanzibar, it is commonly found in shallow lagoons, where it often grows within seagrass beds and/or surrounded by green algae such as Ulva sp. Since seagrasses in Zanzibar have recently been shown to raise the pH of the surrounding seawater during the day, and since calcification rates are sensitive to pH, which changes the saturation state of calcium carbonate, we measured the effects of pH on photosynthetic and calcification rates of this alga. It was found that pH had significant effects on both calcification and photosynthesis. While increased pH enhanced calcification rates both in the light and in the dark at pH >8.6, photosynthetic rates decreased. On the other hand, an increase in dissolved CO₂ concentration to 26 μmol kg⁻¹ (by bubbling with air containing 0.9 mbar CO₂) caused a decrease in seawater pH which resulted in 20% less calcification after 5 days of exposure, while enhancing photosynthetic rates by 13%. The ecological implications of these findings is that photosynthetically driven changes in water chemistry by surrounding plants can affect calcification rates of coralline algae, as may future ocean acidification resulting from elevated atmospheric CO₂.     

An improved model for the calculation of CO2 solubility in aqueous solutions containing Na, K, Ca, Mg, Cl, and SO4

An improved model is presented for the calculation of the solubility of carbon dioxide in aqueous solutions containing Na⁺, K⁺, Ca²⁺, Mg²⁺, Cl⁻, and SO₄²⁻ in a wide temperature–pressure–ionic strength range (from 273 to 533 K, from 0 to 2000 bar, and from 0 to 4.5 molality of salts) with experimental accuracy. The improvements over the previous model [Duan, Z. and Sun, R., 2003. An improved model calculating CO₂ solubility in pure water and aqueous NaCl solutions from 273 to 533K and from 0 to 2000 bar. Chemical Geology, 193: 257–271] include: (1) By developing a non-iterative equation to replace the original equation of state in the calculation of CO₂ fugacity coefficients, the new model is at least twenty times computationally faster and can be easily adapted to numerical reaction-flow simulator for such applications as CO₂ sequestration and (2) By fitting to the new solubility data, the new model improved the accuracy below 288 K from 6% to about 3% of uncertainty but still retains the high accuracy of the original model above 288 K. We comprehensively evaluate all experimental CO₂ solubility data. Compared with these data, this model not only reproduces all the reliable data used for the parameterization but also predicts the data that were not used in the parameterization. In order to facilitate the application to CO₂ sequestration, we also predicted CO₂ solubility in seawater at two-phase coexistence (vapor–liquid or liquid–liquid) and at three-phase coexistence (CO₂ hydrate–liquid water–vapor CO₂ [or liquid CO₂]). The improved model is programmed and can be downloaded from the website http://www.geochem-model.org/programs.htm.     

First observations of jelly-falls at the seafloor in a deep-sea fjord

Faunal communities at the deep-sea floor mainly rely on the downward transport of particulate organic material for energy, which can come in many forms, ranging from phytodetritus to whale carcasses. Recently, studies have shown that the deep-sea floor may also be subsidized by fluxes of gelatinous material to the benthos. The deep-sea scyphozoan medusa Periphylla periphylla is common in many deep-sea fjords in Norway and recent investigations in Lurefjorden in western Norway suggest that the biomass of this jellyfish currently exceeds 50000 t here. To quantify the presence of dead P. periphylla jellyfish falls (hereafter termed jelly-falls) at the deep seafloor and the standing stock of carbon (C) and nitrogen (N) deposited on the seafloor by this species, we made photographic transects of the seafloor, using a ‘Yo-Yo’ camera system during an opportunistic sampling campaign in March 2011. Of 218 seafloor photographs taken, jelly-falls were present in five, which resulted in a total jelly-fall abundance of 1×10^-2 jelly-falls m⁻² over the entire area surveyed. Summed over the entire area of seafloor photographed, 1×10^-2 jelly-falls m⁻² was equivalent to a C- and N-biomass of 13 mg C m⁻² and 2 mg N m⁻². The contribution of each jelly-fall to the C- and N-amount of the sediment in the immediate vicinity of each fall (i.e. to sediment in each 3.02 m² image in which jelly-falls were observed) was estimated to be 568±84 mg C m⁻² and 88±13 mg N m⁻². The only megafaunal taxon observed around or on top of the jelly-falls was caridean shrimp (14±5 individuals jelly-fall⁻¹), and shrimp abundance was significantly greater in photographs in which a jelly-fall was found (14±5 individuals image⁻¹) compared to photographs in which no jelly-falls were observed (1.4±0.7 individuals image⁻¹). These observations indicate that jelly-falls in this fjord can enhance the sedimentary C- and N-amount at the deep-sea floor and may provide nutrition to benthic and demersal faunas in this environment. However, organic enrichment from the jelly-falls found in this single sampling event and associated disturbance was highly localized.