Bioaccumulation study of acrylates in algae (<Emphasis Type="Italic">Chlorella kessleri</Emphasis>) by Py-GC and Py-GC/MS

Acrylate monomers methylmethacrylate (MMA) and cyclohexylmethacrylate (CHMA) bioaccumulation has been determined in aquatic organism, algae (Chlorella kessleri). Algae were collected in amount of 0.4 mg and directly injected to the pyrolytical cell. In algae bodies accumulated monomers were analysed by pyrolysis gas chromatography (Py-GC) and pyrolysis gas chromatography coupled with mass spectrometry (Py-GC/MS). Traces of the accumulated monomers in algae body can be determined after 1-, 2-, 3-weeks of incubation. Maximum content of MMA was determined after 3-week of experiment, contrariwise in the case of CHMA after 2-week exposition. Relationship with pyrolysis temperature has also been studied.     

Determination of total phosphorus and nitrogen in turbid waters by oxidation with alkaline potassium peroxodisulfate and low pressure microwave digestion, autoclave heating or the use of closed vessels in a hot water bath: comparison with Kjeldahl digestion

The evaluation of the use of alkaline peroxodisulfate digestion with low pressure microwave, autoclave or hot water bath heating for the determination of total phosphorus and nitrogen in turbid lake and river waters is described. The efficiency of these digestion procedures were compared to a Kjeldahl digestion procedure with sulphuric acid-potassium sulfate and copper sulfate. The final solution before digestion was 0.045 M in potassium peroxodisulfate and 0.04 M in sodium hydroxide. Procedures were evaluated by the analysis of suspensions of two reference materials, National Institute of Environmental Science, Japan, no. 3 Chlorella and no. 2 pond sediment and natural turbid waters. Best recoveries of phosphorus and nitrogen by microwave heating were obtained when solutions were digested at 95 °C for 40 min. Quantitative recoveries of phosphorus from Chlorella suspensions up to 1000 mg/l were obtained by all three heating procedures, but incomplete recoveries of nitrogen occurred above 20 mg N/l in the digested sample. Good recoveries of phosphorus and nitrogen from suspended sediment suspensions were obtained only from solutions containing <150 mg/l of suspended sediments. Recoveries of phosphorus from phosphorus compounds containing COP and CP bonds added to distilled water were quantitative (94-113%) except for polyphosphates (microwave, 34±8; autoclave, 114±6; water bath, 96±4) and aluminium phosphate (8-23%). Recoveries of nitrogen compounds containing CN bonds added to distilled water were quantitative (94-96%). The analysis of a range of natural turbid water samples by alkaline peroxodisulfate and microwave, autoclave and water bath heating gave similar total phosphorus and nitrogen results. All procedures using alkaline peroxodisulfate underestimate phosphorus concentrations at high suspended sediment concentrations (>150 mg/l) and are only suitable for the analysis of very turbid samples when the turbidity is due to organic matter (algal cells, plant detritus). Underestimation of nitrogen occurs when samples contain more than 20 mg N/l.     

Study of the parameters affecting the binding of metals in solution by Chlorella vulgaris

The ability of Chlorella vulgaris to accumulate heavy metals in solution (Mn, Cr, Ni, Zn and Cu) was investigated. Various parameters (algal biomass, pH and contact time of the algae with the sample) have been studied. Nine mg of algal biomass, pH 8 and 15 min of contact time, with 1 ppm of each metal, were the optimized conditions. At pH 8, the optimum value to rise the maximum binding, a fraction of metals in solution precipitates forming hydroxides. Combining both processes, a chemical–biological system for the removing of metals at ppb levels from the environment is obtained. The simultaneous determination of these five metals was performed by capillary electrophoresis (CE) with a UV/Vis detector.     

Biotransformation of arsenate to arsenosugars by Chlorella vulgaris

Chlorella vulgaris was cultivated in a growth medium containing arsenate concentration of <0.01, 10, 100 and 1000 mg l^?1. Illumination was carried out in 12 h cycles for 5 days. The health status of the culture was monitored by continuous pH and dissolved oxygen (DO) readings. Destructive sampling was used for the determination of biomass, chlorophyll, total arsenic and arsenic species. The chlorophyll a content, the DO and pH cycles were not significantly different for the different arsenate concentrations in the culture. In contrast, biomass production was significantly (p < 0.05) increased for the arsenic(V) treatment at 1000 mg l^?1 compared with 100 mg l^?1. The arsenic concentration in the algae increased with the arsenate concentration in the culture. However, the bioconcentration factor decreased a hundred‐fold with increase of arsenate from the background level to 1000 mg l^?1. The arsenic species were identified by using strong anion‐exchange high‐performance liquid chromatography–inductively coupled plasma mass spectrometry analysis after methanol/water (1 : 1) extraction. The majority (87–100%) of the extractable arsenic was still arsenate; arsenite was found to be between 1 and 6% of total extractable arsenic in the algae. In addition to dimethylarsinic acid, one unknown arsenical (almost co‐eluting with methylarsonic acid) and three different arsenosugars have been identified for the first time in C. vulgaris growing in a culture containing a mixture of antibiotics and believed to be axenic. The transformation to arsenosugars in the algae is not dependent on the arsenate concentration in the culture and varies between 0.2 and 5% of total accumulated arsenic. Although no microbiological tests for bacterial contamination were made, this study supports the hypothesis that algae, and not associated bacteria, produce the arsenosugars. Copyright © 2003 John Wiley & Sons, Ltd.     

Isolation and purification of canthaxanthin from the microalga Chlorella zofingiensis by high-speed counter-current chromatography

Certain microalgae are considered to be a potential source of canthaxanthin, which possesses strong antioxidant and anticancer activities. A high-speed counter-current chromatography (HSCCC) method was developed for the separation and purification of canthaxanthin from the microalga Chlorella zofingiensis. The crude canthaxanthin was obtained by extraction with organic solvents after the microalgal sample had been saponified. Preparative HSCCC, with a two-phase solvent system composed of n-hexane-ethanol-water (10:9:1 v/v), was successfully performed yielding canthaxanthin at 98.7% purity from 150 mg of the crude extract (2.1% canthaxanthin) in a one-step separation. The recovery of canthaxanthin was 92.3%. This was the first report that canthaxanthin was successfully separated and purified from microalgae.     

无机砷影响蛋白核小球藻光合活性的特征研究

采用蛋白核小球藻(Chlorella pyrenoidosa)进行无机砷毒性暴露实验,分析As(Ⅲ)和As(V)对蛋白核小球藻光合活性的影响差异.结果表明:该藻对无机砷具有较强的耐受性,在砷浓度大于37.5mg·L~(-1)暴露条件下,小球藻叶绿素a合成、光系统II(PSII)最大光量子产量(F_v/F_m)、实际光量子产量(Yield)及相关参数与无机砷浓度呈显著负相关,As(Ⅲ)对小球藻光合活性抑制作用显著高于As(V).As(Ⅲ)和As(V)150.0mg·L~(-1)暴露96h对小球藻光系统II影响差异最大,F_v/F_m、Yield、rETR_(max)和线性斜率(α)分别降低了77%,91%,92%,85%和19%,50%,51%,23%.透射电镜进一步表明,小球藻亚细胞结构形态受砷胁迫出现叶绿体片层间空泡化、类囊体基粒片层受挤压、蛋白核缩小及脂质滴.As(Ⅲ)对蛋白核小球藻光系统II(PSII)抑制作用大于As(V),可为深入了解无机砷对淡水微藻光合活性的影响机理提供一定的参考.     

盐度对蛋白核小球藻生长、叶绿素荧光参数及代谢酶的影响

以蛋白核小球藻820为实验材料,研究了3种盐度(15、30、45)对其生长、叶绿素荧光参数和两种代谢酶活性的影响,以了解该小球藻对盐度的适应能力.结果表明,蛋白核小球藻820的生长随盐度增加而变慢;而油脂含量随盐度增加而升高.叶绿素荧光参数中的PSII最大光能转化效率(Fv/Fm)、PSII实际光能转化效率(ΦPSII)、光化学淬灭系数(qP)随盐度升高下降,而非光化学淬灭系数(NPQ)随盐度升高而上升.超氧化物歧化酶(SOD)活性变化大致趋势是低盐和高盐下活性较高,碳酸酐酶(CA)活性则随盐度升高而降低,第5 d时45盐度是30盐度培养的0.43倍.因此,认为高盐一定程度地抑制了蛋白核小球藻的生长、叶绿素荧光参数和CA活性,但是促进了总脂含量和抗氧化酶SOD活性的提高.     

藻类叶绿素荧光对除草剂生物毒性响应特性的研究

以蛋白核小球藻、铜绿微囊藻、斜生栅藻为敏感藻,研究了莠去津、敌稗、敌草隆、灭草松四种除草剂及其混合剂对藻类光合特性的影响规律,实验同时确定了三种微藻荧光对除草剂的最佳响应时间。实验结果表明,除草剂显著降低了藻细胞类囊体膜中光系统Ⅱ的最大光合效率(Fv/Fm)、实际光合效率(Y(Ⅱ))、绝对电子传递速率(ETR)和光化学猝灭系数(qP),而非光化学猝灭系数(qN)呈现上升趋势,400 μg·L-1敌草隆使蛋白核小球藻Fv/Fm参数值下降了41%。实验选用的三种实验藻在四种除草剂单独和混合胁迫下均发生了不同程度的光抑制效应,表现出光合效率、电子传递速率、光合活性的降低和光保护能力的增强,藻类自身具有一定的光保护机制可以降低除草剂的影响。除草剂能够影响藻细胞叶绿素荧光强度,其中灭草松对蛋白核小球藻的影响最为显著,在400 μg·L-1灭草松影响下蛋白核小球藻的叶绿素荧光强度下降了44%。     

叶绿素荧光技术对受Cu~(2+)胁迫藻类暗适应时间的研究

利用叶绿素荧光技术,对受相同浓度Cu2+胁迫的蛋白核小球藻、斜生栅藻、铜绿微囊藻的最佳暗适应时间进行研究.通过对三种供试藻在光照和暗适应时间分别为30s、1min、3min、5min、10min和20min条件下的光合荧光参量进行测定,以光化学淬灭参量值为主要参考依据,结合t检验方法对暗适应时间进行显著性差异分析,结果表明:暗适应条件下三种供试藻的潜在最大量子效率值略有增加,实际量子效率值基本保持不变;蛋白核小球藻和斜生栅藻的光化学淬灭参量值和非光化学淬灭参量值随暗适应时间的延长显著增加;铜绿微囊藻光化学淬灭参量值在光照1min时达到最大,无需进行暗适应,这可能与蓝藻在暗适应时发生状态转换有关;藻类不是暗适应时间越长越好,蛋白核小球藻和斜生栅藻的最佳暗适应时间分别为5min和10min.这将为采用叶绿素荧光技术进一步研究毒物对藻类的胁迫机理提供可靠依据.