Effects of iron electrovalence and species on growth and astaxanthin production of Haematococcus pluvialis

To increase the cell concentration and the accumulation of astaxanthin in the cultivation of Haematococcus pluvialis, effects of different iron electrovalencies (Fe²⁺-EDTA and Fe³⁺-EDTA) and species (Fe-EDTA, Fe(OH)_x^32x and FeC₆H₅O₇) addition on cell growth and accumulation of astaxanthin were studied. Results show that different iron electrovalencies have various effects on cell growth and astaxanthin accumulation of H. pluvialis. Compared with Fe³⁺-EDTA, Fe²⁺-EDTA stimulate more effectively the formation of astaxanthin. The maximum astaxanthin content (30.70 mg/g biomass cell) was obtained under conditions of 18 μmol/L Fe²⁺-EDTA, despite the lower cell density (2.3×10⁵ cell/ml) in such condition. Fe³⁺-EDTA is more effective than Fe²⁺-EDTA in improving the cell growth. Especially, the maximal steady-state cell density, 2.9×10⁵ cell/ml was obtained at 18 μmol/L Fe³⁺-EDTA addition. On the other hand, all the various species of iron (EDTA-Fe, Fe(OH)_x^32x, FeC₆H₅O₇) are capable to improve the growth of the algae and astaxanthin production. Among the three iron species, FeC₆H₅O₇ performed the best. Under the condition of a higher concentration (36 μmol/L) of FeC₆H₅O₇, the cell density and astaxanthin production is 2 and 7 times higher than those of iron-limited group, respectively. The present study demonstrates that the effects of the stimulation with different iron species increased in the order of FeC₆H₅O₇, Fe(OH)_x^32x and EDTA-Fe. Supported by Xiamen Scientific and Technologic Projects (No. 3052Z20031086), Xiamen University Alumni Association Foundation in Singapore, and the First National College Students Innovative Experimental Project     

DYNAMIC CHANGES OF INORGANIC NITROGEN AND ASTAXANTHIN ACCUMULATION IN HAEMATOCOCCUS PLUVIALIS

ＩＮＴＲＯＤＵＣＴＩＯＮＯｖｅｒｔｈｅｐａｓｔｄｅｃａｄｅ ,Ｈａｅｍａｔｏｃｏｃｃｕｓｐｌｕｖｉａｌｉｓｈａｓｂｅｅｎａｓｕｂｊｅｃｔｏｆｎｕｍｅｒｏｕｓｉｎｖｅｓｔｉｇａｔｉｏｎｓｂｅ ｃａｕｓｅｉｔｓｃｏｎｓｐｉｃｕｏｕｓｒｅｄｃｏｌｏｒｐｏｉｎｔｓｔｏｉｔｓｐｏｓｓｉｂｉｌｉｔｙａｓａｈｏｐｅｆｕｌｒｅｇｅｎｅｒａｔｉｖｅｂｉｏｒｅｓｏｕｒｃｅｏｆａｓｔａｘａｎ ｔｈｉｎ .Ｅｘｔｅｎｓｉｖｅｓｔｕｄｙｏｆｃｕｌｔｕｒｅｆａｃｔｏｒｓ (ｓｕｃｈａｓｔｅｍｐｅｒａｔｕｒｅ,ｉｌｌｕｍｉｎａｔｉｏ…     

氮、磷、铁、硅对简单双眉藻生长的影响

用单因子对比实验和正交实验的方法研究了氮、磷、铁和硅对简单双眉藻生长的影响。结果表明:简单双眉藻对于硝酸钠和尿素均能很好地利用,其相对生长常数无显著差异,对柠檬酸铁和三氯化铁的利用亦无显著差异,FeC6H5O7-Fe的适宜浓度为0.1～0.5mg/L;Na2SiO3-Si的适宜浓度为0.8～1.6mg/L。正交实验表明,简单双眉藻氮、磷、铁、硅的最佳浓度(mg/L)配比为6.6∶0.66∶0.1∶0.4。     

Changes of pigments and lipids composition in Haematococcus pluvialis vegetative cell as affected by monochromatic red light compared with white light

Journal of Oceanology and Limnology - Much attention has been paid on studies of astaxanthin accumulation process in Haematococcus pluvialis industry. However, growth of H. pluvialis in motile...     

Determination of Ca,K, Mg and Fe in four fish species by FAAS

A method for determining calcium(Ca),potassium(K),magnesium(Mg) and iron(Fe) in four fish species was optimized and validated.It included microwave mineralization of the samples and subsequent quantification by flame atomic absorption spectroscopy(FAAS) with Zeeman-effect background correction.Using HNO3(65%) and H2O2(33%) as extraction solutions,the optimal conditions of extraction were established as follows:0.5 g of sample mass;microwave time program of 300 W/5 min and 600 W/5 min.The method was free of matrix interferences.The linear correlation coefficients were ≥0.9991,the recovery percentage of analytes was from 99.31% to 103.70% and the RSD(relative standard deviation) was lower than 2.06%.The detection limits obtained were 32.3,43.2,14.0 and 68.6 ng mL-1 for Ca,K,Mg and Fe in FAAS respectively.It is shown that the method is rapid,simple,sensitive and accurate.The method was applied to the studies of digestibility and measurement of these nutrients in samples of fish collected from Norway,Japan and China.     

Effects of Fe and Mn on the growth of a red tide dinoflagellateScrippsiella trochoidea (Stein) Loeblch III

Batch culture experiments were conducted with a red tide dinoflagellateScrippsiella trochoidea (Stein) Loeblch III collected from Jiaozhou Bay, Shangdong, China. Growth rates and oellular Chl—a were measured in media with iron and manganese ion concentrations controlled at different levels using EdTA-trace metal buffer systems. Cell density increased 3.2 times to 6.5 times over the range of lowest (0 mol/L) to highest (10⁻⁵ mol/L) iron and manganese ion concentrations. The range of cell density response was much lower than the range of total available iron and manganese, which was >100—fold that of Fe. This nonlinear relationship indicates that the cells adapt to make more efficient use of iron and manganese under limiting conditions. The cellular Chl—a content maximized after 3 days incubation and then decreased gradually under either iron or manganese limitation conditions. It indicated that the algae gained higher photosynthesis ability when transferred to a new environment. Growth responses to iron and manganese limitation can be both modeled according to the equation of Monod. The half—saturation constant for growth,k, is 4.6×10⁻⁸ mol/L for Fe and 5.1×10⁻⁸ mol/L for Mn. Our results showed that the iron availability in Jiaozhou Bay does not limit the growth ofS. trochoidea. Contribution No. 2831 from the Institute of Oceanology, Chinese Academy of Sciences. Project 9389008 supported by NSFC; Study supported by PDB6.     

Membrane preconcentration of iron in seawater samples and on-site determination in spectrophotometry

A novel method for on-site determination of trace iron was developed using membrane preconcentration and spectrophotometric detection. Fe(Ⅱ)-ferrozine complex was reacted with cetyltrimethylammonium bromide (CTAB) to form a Fe(Ⅱ)-ferrozine-CTAB paired compound, which was collected on a membrane by filtration under vacuum. The membrane was immersed in 2 mL of ethanol-nitric acid and the absorbance of the solution measured for quantitative analysis. Various factors affecting the iron collection and determination were investigated. With different sample preconcentration volumes, the range of determination was broadened to 0.5-120 g/L. The detection limit of this method reached 0.19 g/L and the recoveries were between 97.2 and 109% when the concentration enrichment was about 45. The relative standard deviation (n = 7) was 1.9% for samples containing 10 g/L Fe. Twelve seawater samples were analyzed on-site using the proposed method, and two were also analyzed in inductively coupled plasma mass spectrometry. No significant difference was shown between the two methods by the Student’s t-test. The method has also been used on-site for iron enrichment experiments with phytoplankton and concluded to be simple, accurate and inexpensive.     

ASPECTS OF IRON NUTRITION IN MACROALGAE ULVA PERTUSA (CHLOROPHYTA) UNDER IRON STRESS

ＩＮＴＲＯＤＵＣＴＩＯＮＩｒｏｎｉｓａｎｉｍｐｏｒｔａｎｔｎｕｔｒｉｔｉｏｎａｌｅｌｅｍｅｎｔｉｎａｌｇａｅｊｕｓｔａｓｉｔｉｓｉｎｈｉｇｈｐｌａｎｔｍｅｔａｂｏｌｉｃｐａｔｈｗａｙｓ.Ｐｈｏｔｏｓｙｎｔｈｅｓｉｓｉｓｄｅｐｅｎｄｅｎｔｏｎｔｈｉｓｅｌｅｍｅｎｔｂｅｃａｕｓｅｃｈｌｏｒｏｐｈｙｌｌｓｙｎｔｈｅｓｉｓｒｅｑｕｉｒｅｓｉｒｏｎｎｕｔｒｉｔｉｏｎ .Ｉｒｏｎｉｓａｌｓｏｅｓｓｅｎｔｉａｌｆｏｒｃｙｔｏｃｈｒｏｍｅｓ,ｆｅｒｒｅｄｏｘｉｎ ,ａｎｄＦｅ ｓｐｒｏｔｅｉｎ .Ｉｔｓｂｉｏｃｈｅｍｉ…     

山东省齐河-禹城地区矽卡岩型铁矿磁铁矿元素地球化学特征及其对成矿作用的指示意义

山东省齐河-禹城地区在黄河北厚覆盖区,有明显的航磁异常和重力异常,通过系统的地质勘探工作,在深部揭露了多处铁矿床,为系统掌握研究区磁铁矿成因机制,服务于基础地质和矿产资源调查,利用EMPA对磁铁矿进行主量元素分析,以确定其成因及成矿环境特征.其中TiO2-Al2O3-MgO和(Ca十Al十Mn)-(Ti十V)成因判别图...     

Cell cycles and proliferation patterns in Haematococcus pluvialis

Most studies on Haematococcus pluvialis have been focused on cell growth and astaxanthin accumulation;far less attention has been paid to cell cycles and proliferation patterns.The purpose of this study was to clarify cell cycles and proliferation patterns in H.pluvialis microscopically using a camera and video recorder system.The complicated life history of H.pluvial i s can be divided into two stages:the motile stage and the non-motile stage.All the cells can be classifi ed into forms as follows:motile cell,nonmotile cell,zoospore and aplanospore.The main cell proliferation,both in the motile phase and non-motile phase in H.pluvialis,is by asexual reproduction.Under normal growth conditions,a motile cell usually produces two,sometimes four,and exceptionally eight zoospores.Under unfavorable conditions,the motile cell loses its fl agella and transforms into a non-motile cell,and the non-motile cell usually produces 2,4 or 8 aplanospores,and occasionally 20–32 aplanospores,which further develop into non-motile cells.Under suitable conditions,the non-motile cell is also able to release zoospores.The larger non-motile cells produce more than 16 zoospores,and the smaller ones produce 4 or 8 zoospores.Vegetative reproduction is by direct cell division in the motile phase and by occasional cell budding in the non-motile phase.There is,as yet,no convincing direct evidence for sexual reproduction.     

济宁市翟村铁矿小体重值与磁性铁品位的回归分析

利用济宁市翟村铁矿勘查过程中取得的体重测试值与磁性铁品位数据进行了回归分析,得出回归方程为y=2.74+0.026x,经显著性、精度检验并结合实际实践,证明该一元回归方程具有显著性,预测精度较高。可以用该方程预测翟村铁矿区小体重值。     

ASPECTS OF IRON NUTRITION IN MACROALGAE ULVA PERTUSA （CHLOROPHYTA） UNDER IRON STRESS

Fe, Chlorophyll (Chl) and total nitrogen (TN) content in tissues were measured in Fe-deficient cultures ofUlva. pertusa over a period of 60 days. Photosynthetic carbon fixation rates were studied at the start of and 30 days after Fe-deficiency culture, when the effects of Fe-deficiency on the ultrastructure were also analyzed. The iron content in tissue decreased exponentially during Fe-deficiency (from 726.7 to 31.6 μg/gdw) and simultaneously Chl and TN content declined to 4.35% and 59.9% of their original levels respectively. Maximum carbon fixation rate (50–250 μmol/m² s) under Fe-deficiency decreased significantly compared with the control (p<0.01) and was 13.6 to 0.365 μg C/cm² h. Photosynthesis in Fe-deficient cells became light-saturated at lower irradiance than that in control. Ultrastructural observations of Fe-deficient cells showed reductions in chloroplast number, some degeneration of lamellar organization, an increase in vacuolar area, a decrease in mitochondrial matrix density, and variation in accumulation body number and morphology. During Fe-deficiency, the algae growth rate continued to decline and after 6 weeks of iron deficiency, no further growth was detectable. These suggested that the lower growth rate ofUlva. pertusa under Fe-deficiency could be due mainly to nitrogen utilization and inhibition of photosynthesis. This project No. 39725023 supported by NSFC for Talented Youths and the Project under the Major State Basic Research of China (Grant No. G1999012011).     

SO2-4/Fe2O3固体超强酸催化合成乙酸丁酯

制备固体超强酸SO4 2 - /Fe2 O3,并测定其红外光谱。以乙酸和正丁醇为反应物、固体超强酸SO4 2 - /Fe2 O3为催化剂 ,进行合成乙酸丁酯的研究。实验结果表明 ,固体超强酸SO4 2 - /Fe2 O3在合成乙酸丁酯反应中的催化活性与浓硫酸相当 ,用 0 1mol正丁醇与 0 2mol乙酸反应 ,催化剂用量 0 3g ,回流时间 90min ,乙酸丁酯收率为 63 8%。     

Stability and changes in astaxanthin ester composition from Haematococcus pluvialis during storage

In this paper, we investigated the effects of temperature, oxygen, antioxidants, and corn germ oil on the stability of astaxanthin from Haematococcus pluvialis under different storage conditions, and changes in the composition of astaxanthin esters during storage using high performance liquid chromatography and spectrophotometry. Oxygen and high temperatures （22-25~C） significantly reduced the stability of astaxanthin esters. Corn germ oil and antioxidants （ascorbic acid and vitamin E） failed to protect astaxanthin from oxidation, and actually significantly increased the instability of astaxanthin. A change in the relative composition of astaxanthin esters was observed after 96 weeks of long-term storage. During storage, the relative amounts of free astaxanthin and astaxanthin monoesters declined, while the relative amount of astaxanthin diesters increased. Thus, the ratio of astaxanthin diester to monoester increased, and this ratio could be used to indicate if astaxanthin esters have been properly preserved. If the ratio is greater than 0.2, it suggests that the decrease in astaxanthin content could be higher than 20%. Our results show that storing algal powder from H. pluvialis or other natural astaxanthin products under vacuum and in the dark below 4~C is the most economical and applicable storage method for the large-scale production of astaxanthin from H. pluvialis. This storage method can produce an astaxanthin preservation rate of at least 80% after 96 weeks of storage.     

BACKGROUND STUDY ON NON-RESIDUAL PHASE OF ELEMENTS IN CHANGJIANG AND HUANGHE ESTUARINE SEDIMENTS

Analysis of Zn, Cu, Pb, Co, Cr, Li, Ni, K, Al, Fe extracted by 1 mol/L HCl or 0.5 mol/LHCl/H_2O_2, showed concentrations of Zn, Cu, Pb, Co, Cr, Fe, Ni were significantly correlated with Li, Al,K, and clay. Two methods are used to indicate the background value of the non-residual phase of elementsin sediments, and are the same as the methods used to indicate the background value of totalconcentrations in sediments. The first method uses correlograms and regression equations,the second usesthe mean element concentrations normalized with grain size. Li, Al, K can be used as reference elements to determine the background value of Zn, Cu, Pb, Co,Cr, Ni, Fe, while the clay concentration's correlation with some extractable concentrations can be used tocalculate the background value of the non-residual phase of elements as a percentage of clay concentrationin the sediments. Based on this study, the concept of using the background value of the non-residualphase of elements to compare the pollution level in differ     

Geochemistry of re and trace elements in ferromanganese nodules and their genesis

Trace and RE element geochemistry and genesis have been studied with respect to ferromanganese nodules from the sediments of the Pacific, Atlantic and Indian oceans.     

Strain H<Subscript>2</Subscript>-419-4 of <Emphasis Type="Italic">Haematococcus pluvialis</Emphasis> induced by ethyl methanesulphonate and ultraviolet radiation

Two strains H2-410 and H2-419 were obtained from the chemically mutated survivors of wild Haematococcus pluvialis 2 by using ethyl methanesulphonate (EMS). Strains H2-410 and H2-419 showed a fast cell growth with 13% and 20% increase in biomass compared to wild type, respectively. Then H2-419-4, a fast cell growth and high astaxanthin accumulation strain, was obtained by exposing the strain H2-419 to ultraviolet radiation (UV) further. The total biomass, the astaxanthin content per cell, astaxanthin production of H2-419-4 showed 68%, 28%, and 120% increase compared to wild H. pluvialis 2, respectively. HPLC (High Performance Liquid Chromatography) data showed also an obvious proportional variation of different carotenoid compositions in the extracts of H2-419-4 and the wild type, although no peak of carotenoids appeared or disappeared. Therefore, the main compositions in strain H2-419-4, like its wild one, were free of astaxanthin, monoester, and diester of astaxanthin. The asexual reproduction in survivors after exposed to UV was not synchronous, and different from the normal synchronous asexual reproduction as the mother cells were motile instead of non-motile. Interestingly, some survivors from UV irradiation produced many mini-spores (or gamete?), the spores moved away from the mother cell gradually 4 or 5 days later. This is quite similar to sexual reproduction described by Elliot in 1934. However, whether this was sexual reproduction remains questionable, as no mating process has been observed.     

DISCOVERY OF ORGANIC MOLECULES IN DEEP-SEA IRON COSMIC DUST

A great quartity of cosmic dust(spherules) was found in deep-sea sediments during May to July,1983manganese nodule investigations by the R/V“XIANGYANGHONG 16”in the area 7°-11°N,167°-178°Win the Pacific.Comprehensive study of the cosmic dust and determination with Laser Raman MolecularMicroprobe(LRMM) of the molecules in it showed that besides Fe-Fe,Fe~(3+)-O,Fe-Ni,Al-O,Fe-Obr-Si and Si-Onb,it also contained the organic molecules C-H-O and C-H-S-O,various no fixed form molecules of C,CH_2,CH_3 and volatile molecules CO_2,H_2O,OH~- and H_2S,etc…The study result has very important applications for exploring the origin of interplanetary dust parti-cles and life material.     

Antioxidative capacity and enzyme activity in Haematococcus pluvialis cells exposed to superoxide free radicals

The antioxidative capacity of astaxanthin and enzyme activity of reactive oxygen eliminating enzymes such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) were studied in three cell types of Haematococcus pluvialis exposed to high concentrations of a superoxide anion radical (O₂⁻). The results show that defensive enzymes and astaxanthin-related mechanisms were both active in H. pluvialis during exposure to reactive oxygen species (ROS) such as O₂⁻. Astaxanthin reacted with ROS much faster than did the protective enzymes, and had the strongest antioxidative capacity to protect against lipid peroxidation. The defensive mechanisms varied significantly between the three cell types and were related to the level of astaxanthin that had accumulated in those cells. Astaxanthin-enriched red cells had the strongest antioxidative capacity, followed by brown cells, and astaxanthin-deficient green cells. Although there was no significant increase in expression of protective enzymes, the malondialdehyde (MDA) content in red cells was sustained at a low level because of the antioxidative effect of astaxanthin, which quenched O₂⁻ before the protective enzymes could act. In green cells, astaxanthin is very low or absent; therefore, scavenging of ROS is inevitably reliant on antioxidative enzymes. Accordingly, in green cells, these enzymes play the leading role in scavenging ROS, and the expression of these enzymes is rapidly increased to reduce excessive ROS. However, because ROS were constantly increased in this study, the enhance enzyme activity in the green cells was not able to repair the ROS damage, leading to elevated MDA content. Of the four defensive enzymes measured in astaxanthin-deficient green cells, SOD eliminates O₂⁻, POD eliminates H₂O₂, which is a by-product of SOD activity, and APX and CAT are then initiated to scavenge excessive ROS.     

Geochronology of mineralization and tectonic setting of Baishidong skarn-type iron deposit in Helong area of Yanbian,Jilin Province,NE China

The Baishidong iron deposit is the only skarn-type iron deposit discovered in the study area. According to mineral assemblage and paragenesis, the mineralization can be divided into four metallogenic stages: early garnet-diopside skarn stage, late magnetite-tremolite-epidote skarn stage, early quartz-pyrite-chalcopyrite stage and late quartz-calcite-pyrite stage. Through LA-ICP-MS zircon U-Pb dating of diorite, which is closely related to mineralization, the results show that the weighted average age is 164.6±1.4 Ma, which limits the mineralization time of Baishidong iron deposit during or slightly later than Middle Jurassic. The diorite rocks are rich in sodium(Na_2O/K_2O=1.24--1.76), aluminium(Al_2O_3=17.41%--18.76%), LREE and large-ion lithophile elements(Ba, K and Sr), depleted in HREE and high-field-strength elements(Y, Nb, Ta, P and Ti), and show strongly fractionated patterns(LREE/HREE=6.58--9.93), no apparent Eu anomalies(δEu=0.91--1.13), which shows similar characteristics to island arc or active continental margin arc magma. The zircon εHf(t) values range from-22.6 to 5.9, and the age of the two-stage model(t_(DM2)) is 836--2 633 Ma. Above data combined with the geochemical characteristics, it is indicated that the magma was a mixture of multiple sources, composed of ancient materials and newly formed crust. During the process of evolution and ascending of magma, separation and crystallization occurred, and a large amount of continental crust material was mixed at the same time. Combined with regional tectonic evolution, the formation of this deposit may be related to the westward subduction of the Paleo-Pacific Plate to the Eurasian Plate.