Cryopreservation of Gametophytes of Laminaria japonica （Phaeophyta） Using Encapsulation-Dehydration with Two-Step Cooling Method

Gametophytes of Laminaria japonica were cryopreserved in liquid nitrogen using encapsulation-dehydration with two-step cooling method. Gametophytes cultured at 10℃ and under continuous irradiance of 30 μmol m-2 s-1 for 3 weeks were encapsulated in calcium alginate beads. The beads were dehydrated in 0.4 molL-1 sucrose prepared with seawater for 6 h, desiccated in an incubator set at 10℃ and 70% relative humidity for 4 h, pre-frozen at either -40℃ or -60℃ for 30 min, and stored in liquid nitrogen for ＞24 h. As high as 43% of survival rate was observed when gametophytes were thawed by placing the beads in 40℃seawater and re-hydrated in 0.05 molL-1 citrate sodium prepared using 30‰ NaC1 7 d later. More cells of male gametophytes survived the whole procedure in comparison with female gametophytes. The cells of gametophytes surviving the preservation were able to grow asexually and produce morphologically normal sporophytes.     

A Kinetic Spectrophotometric Method for the Determination of Iron （Ⅲ） in Water Samples

A new kinetic spectrophotometric method has been developed for the determination of iron (Ⅲ). The method is based on the catalytic effect of iron (Ⅲ) on the oxidation of weak acid brilliant blue dye (RAWL) by KIO4 in acid medium. The advantages of the proposed method are that it is sensitive, accurate, rapid, inexpensive, can be operated under room temperature and has a large determination concentration range compared to other techniques. The obtained optimum conditions are pH 3.15, RAWL (200 mgL-1) 5.00mL, Potassium periodate solution (0.01 molL-1) 0.30mL, phenanthroline (0.02molL-1) 1.00mL, reaction temperature 25℃ and reaction time 7 miu. With this method iron could quantitively be determined in the range 0.00-0.02 mgL-1, the detection limit being 4.10×10-10mL-1. The relative standard deviations (RSD) in five replicate determinations for 3 μgL-1 and 5μgL-1 iron (Ⅲ) are 3.1% and 1.9%, respectively. The method has been applied to the determination of iron (Ⅲ) in tap water samples and seawater samples (from the South China Sea), the recovery rates being 98.0% and 100.5%, respectively.     

A study on the flux,speciation, and budget of mercury in controlled experimental ecosystems

The lowest addition of mercury (0.1 ug Hg 1⁻¹) was used in CEEs for research on mercury flux, speciation and budget. The removal behavior of mercury by phytoplankton in water columns of CEEs can be described by first order kinetic equations for total and particulate mercury in the CEE spiked by mercury. The removal rate of mercury in water columns depends on the size and productivity of phytoplanton in a water column to which mercuric ions were added. A 4.4 day half-life time and a 2.8 day half-life time for total and particulate mercury respectively was obtained in diatom bloom. During microflagellate bloom a 30 day total mercury half-life time was estimated with increase of particulate mercury in the water column. The 0.010 ug Hg cm⁻²y⁻¹ mercury flux rate that was attained in the control bag agreed with the values from field measurements in Saanich Inlet where the bags were launched. The proportion of total mercury to dissolved and particulate mercury depended also on the size, productivity, and concentration of mercury in a water column. A more or less constant distribution of mercury species in the control bag was observed as follows: dissolved Hg 0.73, particulate Hg 0.27, inorganic Hg 0.42, dissolved organic Hg 0.31. After spiking with mercury, the particulate mercury rose rapidly and reached to over 70% of the total mercury. The concentration factor of mercury by phytoplankton in the CEEs in the order of 10⁵ was consistent with the results from field measurements in Saanich Inlet. The mercury recovery from the water column, sediment, water with sediment, and the CEE bag walls was only 52.3% of total mercury spiked in the CEE. The losses of mercury by vaporization into the ambient air and diffusion through the wall of the enclosure should be considered. This paper was published in Chinese inOcean. Limn. Sinica,17(4):307–317, 1986.     

THE DENSITIES OF CHANGJIANG ESTUARINE WATERS

The International Eq一lation of State of Seawater,1980 and the PraeticalSalinity Scale,1978 have been adoPted by theUNESCO江CES沼COR八APSOJoint Panel ono‘eanogral,hie Tables and Standards(JPOTS),and endorsed bythese organizati6ns(Miller‘〕and Poisson,1981;Uneseo,1981).Th已new equa-tion and the Praetieal Salinity Seale are to be used for all values Published fromJan .1,1982 .The new equation 15 aeeurate for use in all oeeanie surfaee waters,but eannot be aeeurately aPPlied to…     

Measurement of trace manganese (II) by the catalytic kinetic spectrophotometric method

A new kinetic spectrophotometric method is developed for the measurement of manganese (II) in water. The method is based on the catalytic effect of manganese (II) with the oxidation of weak acid brilliant blue dye (RAWL) by KIO₄ using the Nitrilo triacetic acid (NTA) as an activation reagent. The optimum conditions obtained are 40 mgL⁻¹ RAWL, 1×10⁻⁴molL⁻¹ KIO₄, 2×10⁻⁴ molL⁻¹ Nitrilo triacetic acid (NTA), pH = 5.8, the reaction time of 3.00 min and the temperature of 20.0 °C. Under the optimum conditions, the proposed method allows the measurement of manganese (II) in a range of 0–50.0 ng mL⁻¹ and with a detection limit of down to 0.158 ng mL⁻¹. The recovery efficiency in measuring the standard manganese (II) solution is in a range of 98.5%–102%, and the RSD is in a range of 0.76%–1.25%. The new method has been successfully applied to the measurement of manganese (II) in both fresh water and seawater samples with satisfying results. Moreover, few cations and anions interfere with the measurement of manganese (II). Compared with other kinetic catalytic methods and instrumental methods, the proposed method shows fairly good selectivity and sensitivity, low cost, cheapness, low detection limit and rapidity. It can be applied on boats easily.     

SEAWATER FLUX OF THE SOUTH CHINA SEA

The South China Sea water can be divided according to depth into three boxes by the pycnoclineand a sill.Using a box model with matter balance,the net seawater fluxes were calculated to be317.9×10~4 m~3/s in box Ⅰ for the upper homogeneous layer outflowing to the adjoining oceans;67×10~4 m~3/s in box Ⅲ for the water entering the basin;240×10~4 m~3/s in box Ⅱ for water entering theSouth China Sea.The upward speed of basin water was calculated to be 8.4×10~(-5) cm/s and that ofseawater flowing up along the pycnocline was calculated to be 8.9×10~(-5) cm/s.     

Photochemical oxidation of benzothiophene in seawater

ＩＮＴＲＯＤＵＣＴＩＯＮｐｈｏｔｏｃｈｅｍｉｃａｌｏｘｉｄａｔｉｏｎｏｆｂｅｎｚｏｔｈｉｏｐｈｅｎｅｉｎｓｅａｗａｔｅｒｉｎｏｒｄｅｒｔｏｌｅａｒｎｍｏｒｅａｂｏｕｔｔｈｅｒｅｍｏｖａｌｐａｔｔｅｒｎｏｆｔｈｉｓｐｏｌｌｕｔａｎｔｉｎｍａｒｉｎｅｅｎｖｉｒｏｎｍｅｎｔｓ.Ｂｅｎｚｏｔｈｉｏｐｈｅｎｅｗａｓｃｈｏｓｅｎｆｏｒｔｈｉｓｓｔｕｄｙｓｉｎｃｅｉｔｉｓｔｈｅｓｉｍｐｌｅｓｔ　　Ｔｈｅｈｉｇｈ -ｐｒｅｓｓｕｒｅｍｅｒｃｕｒｙｌａｍｐｕｓｅｄｉｎｔｈｉｓｓｔｕｄｙｃｏｕｌｄｐｒｏｄｕｃｅｌｉｇ…     

Long-term ecological interactions between nutrient and phytoplankton community in the Changjiang estuary

Both nitrate (NO₃⁻) and soluble reactive phosphate (PO₄³⁻) concentration in the freshwater end-member at the mouth of the Changjiang River have increased dramatically since the 1960s. Within the same period in the sea area, with surface salinity>30, NO₃⁻ concentration has shown an obvious increase, PO₄³⁻ has not changed greatly and dissolved reactive silica (SiO₃²⁻) has deceased dramatically. An examination of the elemental ratio of NO₃⁻ to PO₄³⁻ at the mouth of the Changjiang River did not show a systematic trend from the 1960s to 2000s largely because both nutrients increased simultaneously. In comparison, the elemental ratio of dissolved inorganic nitrogen (DIN) to PO₄³⁻ in surface seawater, with salinity>22, has shown a clearly increasing trend. Furthermore, an overall historical change of the SiO₃²⁻:PO₄³⁻ ratio has undergone a reverse trend in this area. Based on the changes of SiO₃²⁻:PO₄³⁻ and DIN:PO₄³⁻ ratios, we can conclude that an overall historical change of SiO₃²⁻:DIN ratio has decreased in this area from the 1950–1960s to 2000s. The argument that phytoplankton productivity in the Changjiang estuary has been enhanced by increasing nutrient input from the riverine transport was supported by these results. A comparative study analyzing the shift of phytoplankton composition from the mid-1980s to 2000s was also made. The results indicated that the average yearly percentage of diatom species in the Changjiang estuary has decreased from 84.6% during 1985–1986 to 69.8% during 2004–2005. Furthermore, the average yearly percentage of diatom abundance in the Changjiang estuary decreased from 99.5% during to 75.5% over the same time period, while the abundance of dinoflagellates has increased dramatically, from 0.7% to 25.4%.     

Alkaline protease production by a strain of marine yeasts

Yeast strain 10 with high yield of protease was isolated from sediments of saltern near Qingdao, China. The protease had the highest activity at pH 9.0 and 45℃. The optimal medium for the maximum alkaline protease production of strain 10 was 2.5 g soluble starch and 2.0 g NaNO3 in 100 mL seawater with initial pH6.0. The optimal cultivation conditions for the maximum protease production were temperature 24.5 ℃, aeration rate 8.0 L min^- 1 and agitation speed 150 r min^-1 . Under the optimal conditions, 623.1 Umg^-1 protein of alkaline protease was reached in the culture within 30 h of fermentation.     

226Ra and 228Ra in the seawater of the western Yellow Sea

1Introduction Therearefourradiumisotopesinnature,ofwhich224Ra,226Raand228Rahavebeenwidelyinvestigated inmarineresearch,butthespecificactivityof223Rais toolowtobemeasured,therefore,itsapplicationsarelimited(Moore,1969;Broeckeretal.,1976;KuandLin,1976;Nozak…     

Determination of Trace Copper （Ⅱ） in Water Samples by Kinetic-spectrophotometry

A new kinetic-spectrophotometric method is proposed for the determination of copper （ Ⅱ ）. The method is based on the catalytic effect of copper （ Ⅱ ） on the oxidation of weak acid brilliant blue dye （RAWL） by hydrogen peroxide. The copper （ Ⅱ ） can be determined spectrophotometrically by measuring the decrease of absorbance of RAWL at λ = 626 nm using the fix-time method. The optimum reaction conditions are as follows： pH 7.20, buffer solution NaOH-KH2PO4, RAWL （200 mgL-1） 5.00 mL, H2O2 （30%） 0.50 mL, reaction temperature 80 ？？ and reaction time 20 min. The linear range of this method is between 0 μg L^-1 and 12 μg L^-1 and the limit of detection is 0.011 μg L-1, the relative standard deviation （RSD） in five replicate determinations for 2 and 8 μg L-1 copper （ Ⅱ ） are 3.2% and 2.3%, respectively. Twenty ions do not interfere in the determination of copper （ Ⅱ ）. The method has been applied satisfactorily to the determination of copper （ Ⅱ ） in freshwater samples （tap water and Yellow River water from Lijin, Shandong, China） and seawater samples （from the South China Sea）, the recovery rates are 98.0%, 102.5% and 96.0%, respectively.     

Effect of NH4-non the pigment content ofLaminaria japonica

Experiments withLaminaria japonica were conducted in Meidao Bay and Pier Bay, Qingdao, China, Nitrogen-starved plants were fertilized intermittently with 7.1 mM NH₄−N solution for 1 hour at 3 day intervals, after which chlorophyll a, fucoxanthin, chlorophyll c and β-carotene contents were analyzed. Photosynthetic and growth rates of plants and nitrogen content of seawater were determined. their contents of chlorophyll a, fucoxanthin, chlorophyll c and β-carotene were 1.65, 0.67, 0.33 and 0.06 mg.dm⁻² respectively, or 2.46, 2.03, 1.86 and 1.81 times those of the controls. The ratio of fucoxanthin to chlorophyll a in nitrogen-enriched plants was lower than that of the controls. Normal growth rate of the plants (2 cm·day⁻¹ in length) were reached when the chlorophyll a content exceeded 0.1 mg·g⁻¹ fresh wt., indicating that chlorophyll a content can serve as an indicator of normal growth. Experimental results show that seawater is regarded as fertile for the normal growth ofLaminaria if the total inorganic nitrogen (including ammonium salt, nitrate and nitrite) content in seawater is about 2 μm, and infertile if less than 1 μm, in which case fertilizer should be applied. Contribution No. 1703 from the Institute of Oceanology, Academia Sinica     

DETERMINATION OF TRACE BENZENE HYDROCARBONS IN JIAOZHOU BAY BY ENRICHMENT AND CAPILLARY COLUMN GAS CHROMATOGRAPHY

Trare amounts of benzene hydrocarbons obtained in Jiaozhou Bay (Qindao) were enriched bysorption on a GDX-102 column and eluted by carbon disulfide. The eluted was concenttaled and then de-temened by capillary column gas cbornatognphy.The contents of virious kinds of benzene hydrocarbons in Jiaozhou Bay coastal water were benzene(22.3-141.6)× 10～(-9)g/L, toluate (15.2-94.0) × 10~(-9) g/L, ethyl benzene(11.8-85.1)×10~(-9) g/L, p -xylene(15.2-78.5) ×10(-9) g/L, m-xylene (10.9-79.4) ×10(-9) g/L, o -xylene (12.4-80.1) x ×10(-9)g/L; iso-propyl(8.4- 73.1) x ×10(-9)g/L, n -propyl (6.9-76.4) ×10(-9) g/L, 1, 3, 5-trimethylbenzene (10.9- 35.9)×10(-9) g/L, 1,2, 4-trimethybenzene (10.0- 38.0)×10(-9) g/L, n - butydriare (8. 1 - 34.6) ×10(-9)g/L. The recovery of benzenehydrocarbons was (85.1 -95.6)%.     

Transfeer of organic pollutants during sewage-seawater mixing

The transfer processes of organic pollutants during sewage-seawater mixing were simulated in three different mixing systems. The release rate of organic matter from sewage particles to seawater was found to be about 0.02–0.04 d⁻¹. The release processes can be simulated by an exchange-equilibrium kinetic model. Degradation of COD was observed in all three mixing systems and can be described by a first order kinetic equation. The degradation rate constantk ranged between 0.04 and 0.07 d⁻¹. These results have provided important parameters for estimation of environmental capacity.     

Heterotrophic bacterial flora in aquaculture area around Xuejiadao

From Oct., 1999 to Oct., 2000, the heterotrophic bacterial flora in the aquaculture area around Xuejiadao was investigated. The result shows that the populations of the heterotrophic bacteria are heavier in summer and autumn than those in winter and spring. The average populations in seawater, sediment, the surface of seaweed and the surface of fish are 1.4×10⁴cfu mL⁻¹, 5.4×10⁶cfu g⁻¹, 1.5×10⁶cfu g⁻¹ and 1.8×10³cfu cm⁻², respectively. A total of 301 strains were isolated, among them 259 were Gram-negative. All the Gram-negative bacteria belong to 13 genera and some genera of Enterobacteriaceae. The communities of bacteria are slightly different among the samples. In the body surface of fish, Genus vibrio is dominant. In the remaining samples, dominant genus is Aeromonas.     

Heterotrophic bacterial floras in industrial area and unpolluted marine environments around Qingdao

From Oct. 1999 to Oct. 2000, the heterotrophic bacterial floras in the industrial marine environment around the Qingdao Power Plant (QPP) and in the unpolluted marine environments were investigated. The results showed that the numbers of the heterotrophic bacteria around QPP were much higher than those in unpolluted environments, and the average numbers in QPP Seawater, QPP Sediment, Unpolluted Seawater and Unpolluted Sediment were 5.4×10⁴cfu(mL)⁻¹, 5.0×10⁵cfug⁻¹, 3.0×10²cfu(mL)⁻¹ and 1.3×10⁵cfug⁻¹ respectively. Totally, 118 strains were isolated from QPP and 99 of them were Gram-negative. One hundred and twenty one strains were isolated from the unpolluted environments and 104 of them were Gram-negative. All the Gram-negative bacteria belonged to 13 genera. The distribution of the bacteria was varied in different marine environments. The results showed that the unpolluted marine environments contained much more Vibrio than seawater and sediment around QPP.     

Controlling mechanisms for the trace metals, (Cu,Zn, Cd,Fe, Co and Ni) in anoxic sea water in Saanich Inlet,British Columbia,Canada

A study was conducted on Cu, Zn, Cd, Fe, Co and Ni levels in Saanich Inlet anoxic seawater. Data on the concentration of these trace metals and H₂S, and other oceanographic parameters were obtained in four cruises. Equilibrium models are presented together with in situ data. The results strongly support the assumption that the solubilities of Cu, Zn, Cd, Fe, Co and Ni are controlled by bisulfide and/or polysulfide complexes. The species of Cu, Zn, Cd and Ni are shown to be Cu(S₄) ₂ ³⁻ and Cu(Sn₄S₅)³⁻, Zn(HS) ₂ ⁰ and ZnHS ₂ ⁻ , Cd(HS) ₂ ⁰ and Ni(HS) ₂ ⁰ , respectively. The solid species controlling Fe²⁺ and Co²⁺ are respectively the pyrohotites FeS and CoS. The data illustrates that thermodynamic equilibrium has been established in the H₂S-controlled seawater of Saanich Inlet, and that equilibrium has not been established in the H₂S−O₂-coexisted seawater of Saanich Inlet. The lack of equilibrium in the H₂S−O₂-coexisted seawater is a result of the flushing or mixing of oxygenated seawater with anoxic water. The new species of trace metals are still in the processes of reduction and precipitation. Contribution No. 1428 from Institute of Oceanology, Academia Sinica     

Study on gametophyte vegetative growth ofUndaria pinnatifida and its applications

When cultured under certain environmental conditions (25°C, light intensity 80 μmol/m²·s, LD 12/12, in enriched seawater medium with 7×10⁻⁴ mol/L NO₃−N, 1.56×10⁻⁴ mol/L, PO₄−P and supplements of other elements like Mn, Fe, I, etc.), male and female gametophytes ofU. pinnatifida kept growing vegetatively and propagated fast at average daily fresh weight increase rate of about 20%. The empirical formulaG _m=G _o·3^m was established to estimate the output of vegetative gametophytes. Vigorous vegetative gametophyte cells began to form reproductive structures (oogonium and spermatangium, when the temperature was lower than 25°C and other environmental factors were kept optimal. The sufficient supply of gametophyte cells provided enough seeds for raisingUndaria sporelings on production scale. Controlled cross-breeding experiments using selected male and female gametophyte clones which increase their cell number by mitosis instead of meiosis were also carried out in vitro. Juvenile sporophytes from the cross-breeding had almost the same length and width increase rates as those of the control. The fact that vegetative gametophytes can be purposely selected, propagated quickly, cross-bred, with the resulting sporophytes having almost the same characteristics leads to a new way to select desiredUndaria strains for long time use without losing the desired economic characteristics. Contribution No. 2696 from the Institute of Oceanology, Chinese Academy of Sciences. Supported by National Natural Science Foundation of China.     

Sea flux of trace metals in Saanich Inlet

The sea fluxes of trace metals, POC, and settled material were studied in anoxic seawater, Saanich Inlet, B. C., Canada with sediment traps. This paper discusses the change of mass fluxes of sediment, trace metals and POC for various seasons and depths. The annual mean of settled material is 1.56 g.m⁻².a⁻¹, 84.6 mg. m⁻².a⁻¹ for POC, 60.0 mg. m⁻².a⁻¹ for Cu, 16.5 mg.m⁻².a⁻¹ for Pb, 189 mg.m⁻².a⁻¹ for Zn, 2.20 mg.m⁻².a⁻¹ for Cd, 699 mg.m⁻².a⁻¹ for Fe, 38.8 mg.m⁻².a⁻¹ for Co, and 84.6 mg.m⁻².a⁻¹ for Ni. The relations between the average fluxes of trace metals and POC, the fluxes of trace metals and settled matter, and the sea fluxes of trace metals and Fe are in linear progression. The resident times of elements as calculated from sea flux, are 1.1 a. for Cu, 0.014 a. for Pb, 0.50 a. for Zn, 3.8 a. for Cd, 0.16 a. for Fe, 0.39 a. for Co, and 1.14 a. for Ni. The order of resident times is as follows: Pb−Fe−Co−Zn−Cu−Ni−Cd. The metal resident times in Saanich Inlet are shorter than the values estimated for the open ocean. It illustrates that the biochemical processes in shallow Saanich Inlet are faster than those in the open ocean, and that debries of plankton and fecal pellets of zooplankton play an important role in vertical transport of organic carbons. Contribution No. 1650 from the Institute of Oceanology, Academia Sinica.     

Bioaccumulation of Pb<Superscript>2+</Superscript> and its effects on growth,morphology and pigment contents of <Emphasis Type="Italic">Spirulina</Emphasis> (<Emphasis Type="Italic">Arthrospira</Emphasis>) <Emphasis Type="Italic">platensis</Emphasis>

A laboratory experiment was conducted to assess the bioaccumulation of Pb²⁺ and its effects on growth, morphology and pigment contents of Spirulina (Arthrospira) platensis. The specimen cultured in Zarrouk liquid medium was treated with various initial metal concentrations (0, 5, 10, 30, 50 and 100 μg mL⁻¹). The growth of S. platensis was adversely affected by Pb²⁺ at high concentrations (30, 50 and 100 μg mL⁻¹). However, at low concentrations (5 μg mL⁻¹), Pb²⁺ could stimulate its growth slightly. The pigment contents (chlorophyll α and β carotene) were decreased in a dose-dependent manner. The highest reductions (67% and 53% respectively in chlorophyll α and β carotene) were observed in 100 μg mL⁻¹ treatment group. The LC₅₀ (96 h) of Pb²⁺ was measured as 75.34 μg mL⁻¹. Apart from a few cases of filament breakages at elevated concentrations (50 and 100 μg mL⁻¹), morphological abnormalities are not specific. Metal bioaccumulation increased with Pb²⁺ concentrations, but decreased with exposure time. The maximum accumulated amount was 188 mg g⁻¹ dry weight. The bioconcentration factor (BCF) reached to a peak at day 2, followed by a gradual reduction for all the exposure concentrations. S. platensis is able to tolerate considerably high Pb²⁺ concentrations. Consequently it can be used as a potential species to remove heavy metal from contaminated waters.