Effect of oil and dispersant on growth and chlorophyll a content of the marine microalga Tetraselmis suecica

Low hydrocarbon concentrations stimulated the growth of Tetraselmis suecica, whereas higher concentrations (200 ppm) inhibited growth. The content of chlorophyll a in this microalga was affected in a similar way. Crude oil had the most marked effects. Dispersant SEAKLIN-101-NT and mixtures of oil and SEAKLIN-101-NT did not show selective toxicity for the microalga, although inhibitory effects could be observed at high concentrations.     

Growth of the oleaginous microalga Aurantiochytrium sp. KRS101 on cellulosic biomass and the production of lipids containing high levels of docosahexaenoic acid

We examined the growth of a novel oleaginous microalga, Aurantiochytrium sp. KRS101, using cellulosic materials as nutrients, and the resultant production of lipids containing high levels of docosahexaenoic acid (DHA). The microalgal strain could grow using either carboxymethylcellulose or cellobiose as a carbon source, and produced lipids containing high levels of DHA (49-58% of total fatty acids). In line with this growth behavior, carboxymethylcellulase and cellobiohydrolase activities were evident in both cell-free lysates and culture broths. Additionally, an industrial cellulosic biomass, palm oil empty fruit bunches (POEFB), a by-product of the palm oil industry, were utilized by the microalgal strain for cell growth and lipid production.     

Influence of laundry detergents on the abundance dynamics and physiological state of the benthic microalga <Emphasis Type="Italic">Attheya ussurensis</Emphasis> (Bacillariophyta) in laboratory culture

This study examines the influence of the detergents “Obychnyi poroshok” and “Ariel” (at 0.1, 1, and 10 mg/l) on the growth and physiological state of the benthic marine microalga Attheya ussurensis. Cell number, growth rate, and oxygen productivity turned out to be the most sensitive characteristics for evaluation of toxic effects, while the pH of the culture medium and the contents of chlorophyll a and carotenoids were more tolerant to detergent. Toxicants in concentrations of 0.1 and 1 mg/l had a weak inhibitory effect on the microalga A. ussurensis; at 10 mg/l, the detergent Ariel exerted a greater inhibitory effect on the microalga than Obychnyi poroshok did.     

Comparative Studies on the Allelopathic Effects of Ulva pertusa Kjellml, Corllina pilulifera Postl et Ruprl, and Sargassum thunbergii mertl O. Kuntze on Skeletonema costatum （Grev.）Cleve

In the present study, we evaluated the allelopathic effects of three macroalgae, namely Ulva pertusa Kjellml, Corallina pilufifera Postl et Ruprl, and Sargassum thunbergii Mertl O. Kuntze, on the growth of the microalga Skeletonema costaturn （Grev.） Creve using culture systems in which the algae coexisted. The effects of the macroalgal culture medium filtrate on S. costatum were also investigated. Moreover, isolated co-culture systems were built to confirm the existence of allelochemicals and preclude growth inhibition by direct contact. The coexistence assay data demonstrated that the growth of S. costaturn was strongly inhibited when fresh tissues, dry powder and aqueous extracts were used; the allelochemicals were lethal to S. costatum at relatively higher concentrations. The effects of the macroalgal culture medium filtrate on the microalga showed both species specificity and complexity. The inhibitory effect of fresh macroalgal tissue and culture medium filtrate on the microalga was due to the alleochemicals released by the macroalgae. The results of the present study show that the allelopathic effects of macroalgae on the microalga are complex. The present study could shed light onto the basis of the interaction between macro- and microalgae.     

Effects of selenium on growth and ultrastructure of the marine unicellular alga <Emphasis Type="Italic">Dunaliella salina</Emphasis> (Chlorophyta)

This study examines the effects of selenium in concentrations of 0.01, 0.5, 1, 5, and 10 mg/liter on the growth and ultrastructure of the microalga Dunaliella salina. Selenium in concentrations of 0.01 and 0.5 mg/liter stimulated cell population growth, while the number of ultrastructural alterations was the same as in the control cells. At a selenium concentrations of 1 mg/liter, cell population growth slightly decreased by the end of the experiment, and there was some increase in the number of cells with damaged organoids and in the number of completely destroyed cells. As well, the excretory function of cell vacuoles was suppressed, and the autophagic activity of these vacuoles was activated to destroy the cytoplasm and nucleus. Concentrations of 5 and 10 mg/liter were toxic to D. salina, suppressing cell population growth and promoting extensive destructive changes. The threshold concentration of selenium for D. salina was 1 mg/liter, which is 1000 times greater than the maximum permissible concentration (MPC). The fact that the microalga was able to survive for several days in this concentration is indicative of its high resistance to selenium.     

IDENTIFICATION AND PHYSIOLOGICAL ASPECTS OF A NOVEL CAROTENOID‐ENRICHED,METAL‐RESISTANT MICROALGA ISOLATED FROM AN ACIDIC RIVER IN HUELVA (SPAIN)1

A heavy‐metal‐resistant, carotenoid‐enriched novel unicellular microalga was isolated from an acidic river in Huelva, Spain. The isolated ribosomal 18S subunit rDNA sequence showed homology with known sequences from green microalgae, the closest sequence (98% homology) belonging to the genus Coccomyxa. The isolated microalga therefore was an up to now uncultured microalga. The microalga was isolated from Tinto River area (Huelva, Spain), an acidic river that exhibits very low pH (1.7–3.1) with high concentrations of sulfuric acid and heavy metals, including Fe, Cu, Mn, Ni, and Al. Electron micrographs show that the microalga contains a large chloroplast with a presence of lipid droplets, an increased number of starch bodies as well as electron‐dense deposits and plastoglobules, the last observed only in iron‐exposed cells. Unlike other acidophile microalgae, the isolated microalga showed high growth rates when cultivated photoautotrophycally (up to 0.6 d^?1) in a suitable culture medium prepared at our laboratory. The growth was shown to be iron dependent. When the microalga is grown in fluidized bed reactors, the high growth rates resulted in unexpectedly high productivities for being a microalga that naturally grows in acidic environments (0.32 g·L^?1·d^?1). The microalga also grows optimally on reduced carbon sources, including glucose and urea, and at an optimal temperature of 35°C. The alga pigment profile is particularly rich in carotenoids, especially lutein, suggesting that the microalga might have potential for antioxidant production, namely, xanthophylls.     

Additive-free harvesting of oleaginous phagotrophic microalga by oil and air flotation

A unique oleaginous phagotrophic microalga Ochromonas danica is poised for effective lipid production from waste. Cell harvesting and dewatering are major costs in making algae-based products. In this work an effective additive-free harvesting method was developed, taking advantage of O. danica’s comparatively more hydrophobic surface and larger size. The algal cells’ partitioning to oil/water interface was evaluated. Recovery by flotation with waste cooking oil was optimized using an L-9 Taguchi orthogonal-array design. Further, additive-free cell collection and concentrating by air flotation was studied for the effects of both physical factors (column dimension, air–stone pore size, sample-to-column volume ratio) and culture properties (pH, culture growth stage, cell concentration, and pure versus impure cultures). The optimized process consistently achieved >90 % recovery in a single stage. 98+ % recovery could be achieved when starting concentrations were >10⁸ cells/ml, or potentially using a two- or multi-stage process for diluter cultures.     

Growth and ultrastructure of the marine unicellular alga <Emphasis Type="Italic">Dunaliella salina</Emphasis> (Chlorophyta) after chronic selenium intoxication

The effects of selenium (0.01, 0.5, 1, 5 and 10 mg/liter) on the growth and ultrastructure of the microalga Dunaliella salina were investigated following its transfer into clean water. Selenium concentrations of 5 and 10 mg/liter were toxic to D. salina, and reinoculation of microalga into clean water did not prevent it from total mortality. When reinoculated from medium with 0.01 mg Se/liter, the cell population density of D. salina was restored in 14 days. The number of ultrastructural alterations in cells was the same as in the control, while the excretory activity of microalga between days 4 and 10 of this experiment was higher. Cell population growth of D. salina transferred from 0.5 and 1 mg Se/liter was lower than in the control. No ultrastructural defects were observed in microalga reinoculated from medium with a selenium concentration of 0.5 mg/liter and the excretion level corresponded to that at 0.01 mg/liter. Various types of ultrastructural damage were found in microalga from medium with 1 mg Se/liter, which was previously reported to be threshold for D. salina; however, the number of cell injuries decreased with increasing time in clean medium. Excretory activity was decreased at the beginning of experiment; but after 7 days, it was restored to the control level. Though there were no ultrastructural alterations in microalgal cells from medium with 0.5 mg Se/liter, we assume that they had molecular defects that could inhibit the cell population growth. The study of microalgae following their reinoculation from medium containing toxicants into clean medium can be a useful method for evaluating algal survival after toxic exposure.     

Effect of Cadmium Ions on Some Biophysical Parameters and Ultrastructure of Ankistrodesmus sp. В-11 Cells

Russian Journal of Plant Physiology - Effects of low concentrations of cadmium ions on growth, photosynthesis, and cell ultrastructure of microalga Ankistrodesmus sp. B-11 were investigated. The...     

Effects of sodium sulfate on the freshwater microalga Chlamydomonas moewusii: implications for the optimization of algal culture media

The study of the microalgal growth kinetics is an indispensable tool in all fields of phycology. Knowing the optimal nutrient concentration is an important issue that will help to develop efficient growth systems for these microorganisms. Although nitrogen and phosphorus are well studied for this purpose, sulfur seems to be less investigated. Sulfate is a primary sulfur source used by microalgae; moreover, the concentration of this compound is increasing in freshwater systems due to pollution. The aim of this study was to investigate the effects of different sodium sulfate concentrations in the culture medium on growth and growth kinetics of the freshwater microalga Chlamydomonas moewusii. Production of biomass, chl content, kinetic equations, and a mathematical model that describe the microalgal growth in relation with the concentration of sodium sulfate were obtained. The lowest concentration of sodium sulfate allowing optimal growth was 0.1 mM. Concentrations higher than 3 mM generated a toxic effect. This work demonstrates that this toxic effect was not directly due to the excess of sulfate ion but by the elevation of the ionic strength. An inhibition model was successfully used to simulate the relationship between specific growth rate and sodium sulfate in this microalga.     

双抗体夹心法检测3-硝基酪氨酸方法的建立及应用

目的：建立一种准确、快速的双抗体夹心酶免疫吸附的方法,以定量检测组织中过氧亚硝基阴离子的水平。方法：分别以小鼠源性抗3-硝基酪氨酸单克隆IgG抗体和兔源性抗3-硝基酪氨酸IgG抗体为包被抗体和检测抗体,采用正交设计方法摸索以上各抗体的浓度,建立定量检测3-硝基酪氨酸（3-NT）的双抗体夹心ELISA法。同时,测定心肌缺血再灌注大鼠心肌组织3-NT的含量。结果：本研究建立的双抗体夹心ELISA法检测3-NT的最低检测下限为0．10ng·ml^-1,具有良好线性关系的检测范围是（0．15-7．50）ng·ml^-1（r^2=0．995）;心肌缺血再灌注组大鼠的心肌组织中的3-NT水平为（1022．42±97．35）ng·mg pro^-1,明显高于假手术组（246．58±56．52ng·mg pro^-1,P〈0．01）。结论：本研究建立的定量检测3-NT的双抗体夹心ELISA法能够方便、准确、快速地检测组织中3-NT的含量,为定量检测组织中ONOO-的水平提供了新的方法。     

Free 3-nitrotyrosine in exhaled breath condensates of children fails as a marker for oxidative stress in stable cystic fibrosis and asthma.

3-Nitrotyrosine (3-NT) is considered as a marker of oxidative stress, which occurs during inflammation. Since 3-NT levels in exhaled breath condensate (EBC) are very low, we applied a specific and sensitive gas chromatography-negative ion chemical ionization-mass spectrometry (GC-NICI-MS) method and high performance liquid chromatography (HPLC) with electrochemical detection for the analysis of free 3-NT in EBC. A total of 42 children (aged 5-17 years) were enrolled in this study, including children with asthma (n=12), cystic fibrosis (n=12), and healthy controls (n=18). Additionally, 14 healthy non-smoking adults (aged 18-59 years) were included. An EcoScreen system was used for the collection of EBC samples. Free 3-NT levels in EBC ranged from 0.54-6.8 nM. Median (interquartile range) concentrations (nM) were similar in all groups: 1.46 (0.97-2.49) in healthy adults, 2.51 (1.22-3.51) in healthy children, 1.46 (0.88-2.02) in children with asthma, and 1.97 (1.37-2.35) in CF children, respectively (p=0.24, Kruskall-Walis test). No difference was found between the children with airway disease and age-matched healthy controls. In healthy subjects, there was no effect of age on 3-NT concentrations. HPLC analyses provided similar concentration ranges for EBC 3-NT when compared with GC-NICI-MS. Our study has clearly demonstrated that free 3-NT in EBC fails as a marker for oxidative stress in children with stable CF and asthma.     

Content of photosynthetic pigments, growth, and cell size of microalga Phaeodactylum tricornutum in the copper-polluted environment

Effects of copper at concentrations of 0.13 and 0.25 mg/l on content of photosynthetic pigments, growth, size characteristics, and cell shape of Phaeodactylum tricornutum (Bacillariophyta) microalga were studied. The inhibitory effect of copper on growth parameters was shown to become stronger with increasing concentration of copper in the growth medium. In spite of considerable retardation of cell growth, the concentrations examined had no effect on the content of photosynthetic pigments though affected some morphometric parameters.     

初始硝酸钠浓度对魏氏真眼点藻的生长、形态和油脂积累的影响

为了确定不同初始氮供应水平对产油微藻魏氏真眼点藻(Eustigmatos vischeri)生长、形态和油脂积累的影响, 本研究通过在改良的BG-11培养基中设置4种不同的初始硝酸钠浓度(17.6、11.7、5.9和3.0 mmol/L)对魏氏真眼点藻(E. vischeri)进行培养。观察结果表明, 魏氏真眼点藻(E. vischeri)的营养细胞为一具裂叶状叶绿体、细胞质中有一红色素体和许多振动颗粒及光滑细胞壁的球形单细胞; 细胞繁殖方式主要是形成二分裂和四分裂的似亲孢子。在低氮条件下, 随着培养时间的延长, 细胞内油体逐步形成, 至培养末期占据细胞的大部分空间, 同时培养物的颜色也由绿色向黄绿色转变, 最终呈橙黄色。实验结果表明, 魏氏真眼点藻(E. vischeri)生物质浓度在17.6 mmol/L组获得最大值为9.14 g/L; 总脂、中性脂和总脂肪酸三者占干重的含量随着初始硝酸钠浓度的降低而升高, 在3.0 mmol/L组获得最高值, 分别为60.81%、56.59%和53.47%; 三者的单位体积产率均在5.9 mmol/L组获得最高值, 分别为0.24、0.21和0.20 g/(Ld); 主要脂肪酸组成为棕榈酸(C16:0)、棕榈油酸(C16:1)、油酸(C18:1)和二十碳五烯酸(C20:53, EPA), 其中棕榈油酸的含量最高。上述研究表明, 魏氏真眼点藻(E. vischeri)是一株适合于生产生物柴油和长链不饱和脂肪酸EPA的高产油微藻。       

成都地区天然水域中异养微藻的分离及其特性研究

采用添加不同浓度和种类抑菌剂的平板进行多次平板划线分离和三角瓶培养,从成都地区四川大学望江及江安校区附近水域中分离得到了一株可在无光照条件下异养生长的微藻,经初步鉴定为小球藻并编号为Chlorella SCU-01.在此基础上初步研究了该藻株在不同葡萄糖起始浓度下的异养生长及色素合成特性.     

Hepatic macromolecular covalent binding of mononitrotoluenes in Fischer-344 rats

The mononitrotoluenes are important industrial chemicals which display isomeric specificity in their ability to induce hepatic DNA excision repair in Fischer-344 rats. Covalent binding of the structurally related hepatocarcinogen, 2,6-dinitrotoluene, to hepatic DNA is markedly decreased by prior administration of the sulfotransferase inhibitors pentachlorophenol (PCP) and 2,6-dichloro-4-nitrophenol (DCNP). The objectives of this study were to determine whether hepatic macromolecular covalent binding of the mononitrotoluene isomers differed and to determine whether covalent binding of the mononitrotoluenes to hepatic DNA in vivo was decreased by inhibitors of sulfotransferase. Male Fischer-344 rats were given a single oral dose of [ring-U-14C]-2-, 3- or 4-nitrotoluene (2-, 3- or 4-NT) and killed at various times thereafter. Livers were removed and analyzed for total and covalently bound radiolabel. Maximal concentrations of total radiolabel were observed between 3 and 12 h after the dose, and there were no large differences among the 3 isomers in peak concentrations achieved. Covalent binding to hepatic macromolecules was maximal 12 h after administration for all three isomers. Thereafter, concentrations of covalently bound 2-NT-derived material were always 2-6 times higher than those of 3- or 4-NT-derived material. When DNA was isolated from livers of rats given the mononitrotoluenes 12 h previously, only 2-NT was observed to covalently bind at concentrations above the limits of detection of the assay. The covalent binding of 2-NT, but not that of 3- or 4-NT, to both total hepatic macromolecules and DNA was markedly decreased by prior administration of either PCP or DCNP. Covalent binding to hepatic DNA was decreased by greater than 96%. The results of this study correlate well with studies which have demonstrated that 2-NT, but not 3- or 4-NT, induces DNA excision repair. Furthermore, they suggest that 2-NT, like the hepatocarcinogen 2,6-dinitrotoluene, requires the action of sulfotransferase for its conversion to a species capable of covalently binding to hepatic DNA.     

Artefacts in HPLC Detection of 3-Nitrotyrosine in Human Brain Tissue

Abstract: An HPLC method was used for quantification of 3-nitrotyrosine (3-NT) in human postmortem brain tissue. A peak with similar retention time to 3-NT was detected in brain tissue from patients with Parkinson's disease, Huntington's chorea, multiple system atrophy, and Alzheimer's disease but not in control tissue. The peak was lost on reduction with dithionite, a criterion often used to identify 3-NT. Tissue from the same neurodegenerative diseases was analysed by HPLC using a photodiode array detector in series with an amperometric electrochemical detector, but the peak was found not to be 3-NT. The absorbance spectrum, fragmentation pattern on mass spectroscopy, and electrochemical profile of this peak do not match authentic 3-NT. A search of the mass spectroscopy databases failed to reveal its identity. The presence of this closely eluting, dithionite-reducible peak could confound analysis of human tissues for 3-NT. In vitro experiments showed that high concentrations of peroxynitrite were needed to achieve detectable levels of 3-NT in human brain tissue.     

Productivity and selective accumulation of carotenoids of the novel extremophile microalga <Emphasis Type="Italic">Chlamydomonas acidophila</Emphasis> grown with different carbon sources in batch systems

Cultivation of extremophile microorganisms has attracted interest due to their ability to accumulate high-value compounds. Chlamydomonas acidophila is an acidophile green microalga isolated by our group from Tinto River, an acidic river that flows down from the mining area in Huelva, Spain. This microalga accumulates high concentrations of lutein, a very well-known natural antioxidant. The aim of this study is to assess use of different carbon sources (CO₂, glucose, glycerol, starch, urea, and glycine) for efficient growth of and carotenoid production by C. acidophila. Our results reveal that growth of the microalga on different carbon sources resulted in different algal biomass productivities, urea being as efficient as CO₂ when used as sole carbon source (~20 g dry biomass m^–2 day^–1). Mixotrophic growth on glucose was also efficient in terms of biomass production (~14 g dry biomass m^–2 day^–1). In terms of carotenoid accumulation, mixotrophic growth on urea resulted in even higher productivity of carotenoids (mainly lutein, probably via α-carotene) than obtained with photoautotrophic cultures (70% versus 65% relative abundance of lutein, respectively). The accumulated lutein concentrations of C. acidophila reported in this work (about 10 g/kg dry weight, produced in batch systems) are among the highest reported for a microalga. Glycerol and glycine seem to enhance β-carotene biosynthesis, and when glycine is used as carbon source, zeaxanthin becomes the most accumulated carotenoid in the microalga. Strategies for production of lutein and zeaxanthin are suggested based on the obtained results.     

Effects of Temperature and pH on Cell Growth of a Flocculating Microalga (Chlorella vulgaris) JSC-7

微藻是可广泛用于健康食品及水产养殖的饵料,同时,微藻细胞内积累的油脂可作为可再生生物燃料,因此微藻的生长和代谢受到广泛关注。温度和pH对微藻的生物量积累有很大影响,考察不同温度和pH条件下微藻细胞的生长有助于寻找最佳的条件进行微藻的培养。自絮凝小球藻JSC-7（Chlorella vulgaris JSC-7）可实现自沉降采收,有利于降低微藻生产成本,优化其生长条件对更好地利用该微藻具有重要意义。考察了温度（22∽40℃）及pH（6.0∽10.0）对其细胞生长、叶绿素含量和油脂产量的影响。在所选取的温度及pH范围内,JSC-7细胞均可生长,显示该藻种可以适应广泛的温度和pH条件。适合细胞生长的温度依次为31℃〉28℃〉35℃〉25℃,pH依次为7.0〉8.0〉6.0。pH 8.0时生物量和油脂的积累量最多,说明该藻株在弱碱条件下更适合生长和产油。当温度为31℃、pH为7.0时,可获得最高的生长量（OD690=0.941）、叶绿素含量（19 mg/L）及油脂产量（39.07%/克干重）。     

The transmembrane domain of CEACAM1-4S is a determinant of anchorage independent growth and tumorigenicity

CEACAM1 is a multifunctional Ig-like cell adhesion molecule expressed by epithelial cells in many organs. CEACAM1-4L and CEACAM1-4S, two isoforms produced by differential splicing, are predominant in rat liver. Previous work has shown that downregulation of both isoforms occurs in rat hepatocellular carcinomas. Here, we have isolated an anchorage dependent clone, designated 253T-NT that does not express detectable levels of CEACAM1. Stable transfection of 253-NT cells with a wild type CEACAM1-4S expression vector induced an anchorage independent growth in vitro and a tumorigenic phenotype in vivo. These phenotypes were used as quantifiable end points to examine the functionality of the CEACAM1-4S transmembrane domain. Examination of the CEACAM1 transmembrane domain showed N-terminal GXXXG dimerization sequences and C-terminal tyrosine residues shown in related studies to stabilize transmembrane domain helix-helix interactions. To examine the effects of transmembrane domain mutations, 253-NT cells were transfected with transmembrane domain mutants carrying glycine to leucine or tyrosine to valine substitutions. Results showed that mutation of transmembrane tyrosine residues greatly enhanced growth in vitro and in vivo. Mutation of transmembrane dimerization motifs, in contrast, significantly reduced anchorage independent growth and tumorigenicity. 253-NT cells expressing CEACAM1-4S with both glycine to leucine and tyrosine to valine mutations displayed the growth-enhanced phenotype of tyrosine mutants. The dramatic effect of transmembrane domain mutations constitutes strong evidence that the transmembrane domain is an important determinant of CEACAM1-4S functionality and most likely by other proteins with transmembrane domains containing dimerization sequences and/or C-terminal tyrosine residues.