Optimization of culture conditions and medium composition for the marine algicidal bacterium Alteromonas sp. DH46 by uniform design

Harmful algal blooms (HABs) have led to extensive ecological and environmental issues and huge economic losses. Various HAB control techniques have been developed, and biological methods have been paid more attention. Algicidal bacteria is a general designation for bacteria which inhibit algal growth in a direct or indirect manner, and kill or damage the algal cells. A metabolite which is strongly toxic to the dinoflagellate Alexandrium tamarense was produced by strain DH46 of the alga-lysing bacterium Alteromonas sp. The culture conditions were optimized using a single-factor test method. Factors including carbon source, nitrogen source, temperature, initial pH value, rotational speed and salinity were studied. The results showed that the cultivation of the bacteria at 28°C and 180 r min^?1 with initial pH 7 and 30 salt contcentration favored both the cell growth and the lysing effect of strain DH46. The optimal medium composition for strain DH46 was determined by means of uniform design experimentation, and the most important components influencing the cell density were tryptone, yeast extract, soluble starch, NaNO₃ and MgSO₄. When the following culture medium was used (tryptone 14.0g, yeast extract 1.63g, soluble starch 5.0 g, NaNO₃ 1.6 g, MgSO₄ 2.3 g in 1L), the largest bacterial dry weight (7.36 g L^?1) was obtained, which was an enhancement of 107% compared to the initial medium; and the algal lysis rate was as high as 98.4% which increased nearly 10% after optimization.     

Influence of light availability on the specific density,size and sinking loss of Anabaena flos-aquae and Scenedesmus obliquus

Harmful algal blooms in eutrophic waters pose a serious threat to freshwater ecosystems and human health. In-situ light availability control is one of the most commonly used technologies to suppress algae in lakes and reservoirs. To develop a better understanding of the effects of light on algal growth, specific density, colony size and sinking loss, Anabaena flos-aquae(cyanobacteria) and Scenedesmus obliquus(green algae) were evaluated in varying light scenarios. The results showed that the specific density and colony size of these two species varied during growth, and there were obvious differences among the light scenarios. At the end of exponential phase, S. obliquus incubated under light-limited condition maintained a higher specific density and formed larger aggregates, whereas A. flos-aquae formed a longer filament length. Both species exhibited higher sinking loss rates with lower light availability. These results implied that the sinking loss rate was not always constant but should be considered as a variable response to the change of light availability, and in-situ light availability control might result in a significant increase of the sinking loss of algae due to the change of size and specific density, thereby further affecting the algal biomass in the water column.     

Production of DMS and DMSP in different physiological stages and salinity conditions in two marine algae

Dimethylsulfide(DMS) and dimethylsulfoniopropionate(DMSP) production by Scrippsiella trochoidea and Prorocentrum minimum was investigated to characterize the effects of physiological stage and salinity on DMS and DMSP pools of these two marine phytoplankton species.Axenic laboratory cultures of the two marine algae were tested for DMSP production and its conversion into DMS.The results demonstrated that both algal species could produce DMS,but the average concentration of DMS per cell in S.trochoidea(12.63 fmol/L) was about six times that in P.minimum(2.01 fmol/L).DMS and DMSP concentrations in algal cultures varied significantly at different growth stages,with high release during the late stationary growth phase and the senescent phase.DMS production induced by three salinities(22,28,34) showed that the DMS concentrations per cell in the two algal cultures increased with increasing salinity,which might result from intra-cellular DMSP up-regulation with the change of osmotic stress.Our study specifies the distinctive contributions of different physiological stages of marine phytoplankton on DMSP and DMS production,and clarifies the influence of salinity conditions on the release of DMS and DMSP.As S.trochoidea and P.minimum are harmful algal bloom species with high DMS production,they might play an additional significant role in the sulfur cycle when a red tide occurs.     

盐度对威氏海链藻生长和生化指标的影响

【目的】研究威氏海链藻在不同盐度条件下的生长和生化组分变化。【方法】设置盐度梯度为5、15、25、35、45,采用血细胞计数板计数法、热乙醇提取法、尼罗红染色法、苯酚硫酸法和考马斯亮蓝法,分别测定威氏海链藻在不同盐度下的生物量、叶绿素a、类胡萝卜素、中性脂质、总糖和蛋白质含量。【结果】盐度对威氏海链藻细胞密度影响显著(P<0.05),当盐度25和盐度35时,培养8 d细胞密度最高,均为2.0×10⁶ mL^-1;盐度对威氏海链藻叶绿素a和类胡萝卜素含量有显著影响(P<0.05),当盐度15时,细胞叶绿素a和类胡萝卜素含量均最高,分别为3.4814μg/mL(4 d)、2.3549μg/mL(8 d)。盐度对威氏海链藻可溶性总糖含量有显著影响(P<0.05),当盐度15时,8 d时可溶性总糖积累含量最高,为123μg/mL。当盐度35时,6 d时可溶性蛋白积累含量最高,为142.9μg/mL,且显著高于其他实验组(P<0.05)。盐度45时,8 d时中性脂含量最高,显著高于其他实验组(P<0.05)。【结论】盐度为15~35时,威氏海链藻生长速度较快,生化组成较为稳定,盐度15时最有利于藻细胞叶绿素a和类胡萝卜素、总糖的合成;盐度25和盐度35时,藻细胞平均增长速度较快,盐度35时最有利于藻细胞可溶性蛋白积累。     

L波段对不同盐类土壤的微波介电特性分析

 土壤盐碱化是世界面临的一大难题。微波遥感探测土壤具有优势,L波段对土壤含水量和含盐量较为敏感。为了分析不同盐类对土壤介电特性的影响,本文利用L波段(1.43 GHz)微波谐振腔测量了我国盐碱土中普遍存在的3种盐类(NaCl,Na₂SO₄,NaHCO₃)土壤样品的介电常数,研究了土壤介电常数与含水量、含盐量和盐种类的关系。得出结论:实部主要由土壤的含水量决定,含盐量对实部的影响无明显规律;虚部值随含水量和含盐量的增加而增大;3种盐类土壤介电常数虚部随含盐量变化呈现不同的递增趋势,在含盐浓度一定的条件下,虚部值是ε″ _NaCI>ε″ _NaHCO3>ε″ _Na2SO4;在某一含盐量点3种盐类土壤虚部随含水量变化的斜率不同,以此能将3种盐类区分。该研究为建立含不同盐类土壤介电常数模型和微波遥感监测土壤盐碱化提供了实验的依据。     

Effect of dietary potassium on growth,nitrogen metabolism,osmoregulation and immunity of pacific white shrimp (Litopenaeus vannamei) reared in low salinity seawater

An 8 weeks feeding experiment was conducted to determine the effect of dietary potassium on the growth and physiological acclimation of Pacific white shrimp(Litopenaeus vannamei) reared in diluted seawater(salinity 4). Six semi-purified practical diets containing 0.59, 0.96, 1.26, 1.48, 1.74, and 2.17+ g potassium K per 100 g diet were formulated, respectively. The survival and feed conversion rate did not show significant difference among groups of shrimps given these diets(P0.05). The shrimps fed the diets containing 0.96-1.48 g K+ per 100 g diet gained the highest weight, specific growth rate, and protein efficiency ratio. Their ammonium-N excretion rate as well as hemolymph concentration of Na+ and Cl- were significantly lower than those of the control(P0.05), but a reverse trend was observed for their gill Na+/K+-ATPase. Moreover, the shrimps fed with 1.48 g K+ per 100 g diet were the highest in hemolymph urea level, and the phenoloxidase and lysozyme activities were significantly higher than those of the control(P0.05). The growth and physiological response of the test shrimps suggested that diet containing 1.48 g K+ per 100 g diet improved the growth of L. vannamei in low-salinity seawater, and enhanced the physiological acclimation of the organism.     

盐度突变对青蛤(Cyclinasinensis)鳃Na~+/K~+-ATPase活性的影响

研究了盐度23下青蛤(Cyclina sinensis)鳃、斧足、外套膜和内脏团中Na+/K+-ATPase的活性差异,并以活性高的鳃组织为对象,对青蛤实施了低盐(23→16)和高盐(23→36)突变的胁迫,监测了鳃组织中Na+/K+-ATPase活性的动态变化。结果表明,盐度23下青蛤鳃组织中Na+/K+-ATPase的活性显著高于其他组织;在盐度突降胁迫下,鳃组织中Na+/K+-ATPase的活性于前16 d逐渐升高,第17d起显著升高(p<0.05),第18 d达到峰值,于第20 d回落,但仍高于起始水平;在盐度骤升胁迫下,鳃组织中Na+/K+-ATPase的活性于前13d逐渐降低,第14 d起显著降低(p<0.05),于第15 d达到最低值,继而回升至平稳水平,但仍低于起始水平。青蛤不同盐度胁迫下其鳃组织中Na+/K+-ATPase活性的响应方式不同,低盐时活性增加,而高盐时活性下降;此外,青蛤在盐度胁迫下对自身渗透压的调节则需较长时间。     

Effects of iron and humic acid on competition between Microcystis aeruginosa and Scenedesmus obliquus revealed by HPLC analysis of pigments

Iron is a vital micro nutrient for growth of bloom-forming Microcystis aeruginosa and competition with other algae,and its availability is affected by humic acid.The effect of iron and humic acid on growth and competition between M.aeruginosa and Scenedesmus obliquus was assessed.The results showed the growth of M.aeruginosa and S.obliquus in mono-cultures was inhibited by humic acid at low iron concentrations(0.01 mg/L);the maximum inhibition ratios were 67.84% and 38.31%,respectively.The inhibition of humic acid on the two species was significantly alleviated when iron concentrations were1.00 mg/L,with the maximum inhibition rate reduced to 5.82% for M.aeruginosa and to 23.06% for S.obliquus.S.obliquus was the dominant species in mixed cultures,and the mutual inhibition between M.aeruginosa and S.obliquus at low iron concentration was greater than that at high iron concentration.The inhibition of S.obliquus on M.aeruginosa was reduced at low iron concentrations;it increased at high iron concentrations,as concentrations of humic acid rose.     

Effect of soil salinity on cold tolerance of mangrove Kandelia candel

Analysis of cold tolerance on mangroveKandelia candel leaf growing in different soil salinity along Jiulong River Estuary in South China showed that the cold tolerance decreased as the increase of soil salinity. The lethal temperatures ofK. candel leaf were −10.4, −9.9 and −8.6 °C in Liaodong, Baijiao and Aotou, respectively. Under 1–2°C cold stress treatment on detached leaves ofK. candel, their caloric value gradually decreased, while electrolyte leakage gradually increased. The leaf's caloric value and electrolyte leakage in Aotou with higher soil salinity varied more largely than those in Liaodong with lower soil salinity. InK. candel leaf, total water content lowered a little, bound water content rose significantly and free water content dropped significantly with duration of cold stress. At the same time, reduction sugar, soluble sugar and starch content gradually decreased and sucrose content gradually increased. Bound water, free water and sucrose content inK. candel leaf from Aotou with higher soil salinity changed more slowly than those from Liaodong with lower soil salinity, but reduction sugar, soluble sugar and starch content inK. candel leaf from Aotou had faster variations than those from Liaodong. These data indicated that soil salinity can reduce cold tolerance ofK. candel leaf by increasing negative effect of salt ions in cell membrane, inhibiting variations of water content, and aggravating consumption of material and energy. Supported by the Found for Outstanding Young Teacher, the Ministry of Education, China.     

几种无机盐对隐甲藻生长和DHA产量的影响

研究了3种无机盐对隐甲藻(Crypthecodinium cohnii ATCC30556)生长及DHA产量的影响.结果表明隐甲藻可在NaCl为唯一无机盐的培养基中生长;培养基中NaCl质量浓度为6 g/L,Mg2+浓度为13.23 mmol/L,PO43-浓度为7.19 mmol/L可满足藻细胞生长和DHA积累的需求,此时隐甲藻有着最大的生物量和DHA产量,分别为3.31 g/L和499.93 mg/L.     

Effect of high salinity on cell growth and protein production of Antarctic ice microalgae Chlamydomonas sp. ICE-L

Antarctic ice microalgae Chlamydomonas sp. ICE-L can survive and thrive in Antarctic sea ice. In this study, Chlamydomonas sp. ICE-L could survive at the salinity of 132‰ NaCl. SDS-PAGE showed that the density of 2 bands (26 and 36 kD) decreased obviously at the salinity of 99‰ NaCl compared to at the salinity of 33‰ NaCl. The soluble proteins in Chlamydomonas sp. ICE-L grown under salinity of 33‰ and 99% NaCl were compared by 2-D gel electrophoresis. After shocking with high salinity, 8 protein spots were found to disappear, and the density of 28 protein spots decreased. In addition, 19 protein spots were enhanced or induced, including one new peptide (51 kD). The changes of proteins might be correlated with the resistance for Chlamydomonas sp. ICE-L to high salinity.     

Effects of Nutritional Factors on the Growth and Heterotrophic Eicosapentaenoic Acid Production of Diatom Nitzschia laevis

The effects of several nutritional factors on the growth and eicosapentaenoic acid (EPA) production of diatom Nitzschia laevis were studied. 4 LDM (quadrupled concentration of the nutrient salt) was the optimal concentration of nutrient salt for the growth and EPA production ofN. laevis. The growth ofN. laevis was inhibited when the glucose concentration was either lower than 10gL-1 or higher than 15gL-1. Both sodium nitrate and urea were good nitrogen sources for the growth and EPA production, while ammonium chloride seriously decreased the dry cell weight (DW) and the EPA content. Silicate seriously influenced the growth of N. laevis. The maximum DW of 2.34gL-1 was obtained in the presence of 150 mgL-1 Na2SiO3·9H2O. The EPA content remained almost the same when the silicate concentration was lower than 150 mgL-1; however, higher silicate concentrations resulted in a steady de- crease of EPA content. Low medium salinity (≤29) did not seem to influence the DW of N. laevis, and high salinity resulted in a decrease of DW. The highest EPA content (4.08%) and yield (110 mgL-1) were observed at the salinity of 36 and 29, respectively.     

Soluble species in aerosols collected on the route of the Second Chinese National Arctic Research Expedition

Aerosol samples are collected on the route of the Second Chinese NationalArctic Research Expedition from July 15 to September 28,2003.The concentration ofwater soluble ions(Na~+,NH_4~+,Ca~(2+),Cl~-,MSA,SO_4~(2-)and so on)are ana-lyzed.By correlation analysis,the ions can be divided into three groups:(1)Na~+,Mg~(2+),K~+,Ca~(2+),Cl~-,SO_4~(2-),mainly from sea salt;(2)NH_4~+,NO_3~-,markedlyfrom coastal regions of the continents;(3)Acetate,MSA,C_2O_4~(2-),from othersources.Marine aerosols are the dominant origin,Cl~- and Na~+ are the most domi-nant anion and cation,respectively and these two ions(Na~++Cl~-)account for 55.6% of the total aerosol loading.The mean equivalence ratio of NH_4~+/SO_4~(2-)is 0.45,we suggest that ammonium and sulfate exist mainly as NH_4HSO_4.The concentrationof NO_3~- shows three different patterns on the route of expedition:Japan Sea with me-dian value of 15.2 neq·m~(-3);Sea of Okhostk and Bering Sea of 1.8 neq·m~(-3)andArctic Ocean of 0.4 neq·m~(-3).Bering Sea is a high MSA area.Through the com-parison of the concentration of main water soluble ions between the First and SecondChinese National Arctic Research Expedition,the variation matches each other.     

Effects of light quality on growth rates and pigments of Chaetoceros gracilis (Bacillariophyceae)

Light is an important parameter in algal culturing. In this work, the effects of different light qualities on growth of the marine diatom Chaetoceros gracilis are evaluated. The cells were cultured under light quantum flux density of 60 µmol photons/(m2·s) with nine light qualities: LED red light (LR), LED blue light (LB), LED red plus LED blue light (LR+LB), LED white light (LW), fluorescent white light (FW), and combinations of LW and FW lights with increased proportions of red or blue light (FW+LR, FW+LB, LW+LR, and LW+LB). Blue light promoted the growth of C. gracilis largely. Three light qualities, FW+LR, LW+LR, and LR, resulted in the lowest growth rate. Both chlorophyll and carotenoids reached the highest level under LB and the lowest level under LR among monochromatic light sources; however, increasing of the proportion of blue or red light in the white light induced an increase of the chlorophyll and carotenoid concentrations. Light absorption ability of microalgae was critical for the growth of these organisms, as we observed a positive correlation between the extinction coefficient at different wavelengths and the specific growth rate. These findings are of importance to improve the culturing of this alga in bioreactors.     

潍坊北部地区土壤盐分光谱特征分析

为提高海水入侵地区土壤盐分的监测效率,以潍坊北部地区作为研究区,利用高光谱测量技术野外实地测量,获得高光谱数据,并对该区域土壤盐分高光谱数据进行分析,获得土样盐分含量相关数据,与光谱数据结合分析。通过分析各类离子与全盐量之间的相关关系,分析潍北地区土壤盐分的离子构成与离子的相关来源。对高光谱原始测量数据进行包络线去除、变换,选取特征波段、一阶二阶微分变换等提高模型精度,建立高光谱数据与土壤盐分数据之间的逐步回归模型,并对模型进行精度检验,对比各类模型检验结果,分析各类离子对高光谱数据的响应效果。其中,光谱数据一阶微分变换后的逐步回归模型对土壤中K+Na+估算精度较高,R2=0.894且均方根误差最小,精度较高,可为该地区的高光谱土壤盐分监测提供有力的数据分析结果。     

Effect of polysaccharide from Enteromorpha prolifera on maize seedlings under NaCl stress

In this study,a polysaccharide from Enteromorpha prolifera(EP)was extracted and its effect on maize seedlings under NaCl stress was investigated.Firstly,the components and structure of the EP were determined.We found that EP is a sulfated polysaccharide of high-molecular weight(Mw,1 840 KDa)heteropoly saccharides and the main mono saccharide is rhamno se.The poly saccharide was applied to explore its effect on the growth of maize seedlings and its defense response under a salt stress.The results show that EP could promote the growth of maize seedlings under the salt stress.In addition,EP was shown able to significantly regulate membrane permeability and adjustment of osmotic substances such as soluble protein,soluble sugar,and proline,antioxidant enzymes containing superoxide dismutase,catalase,peroxidase,and ascorbate peroxidase.Therefore,EP is an effective salt-resistant substance for the growth of maize seedlings under NaCl stress.     

Effects of oxytetracycline dihydrate and sulfamethoxazole on Microcystis aeruginosa and Chlamydomonas microsphaera

The increasing use of pharmaceuticals has become a major environmental issue in China.The presence of antibiotics in water may have deleterious effects on non-target aquatic organisms such as microalgae.In this study,a cyanobacterium and an alga species in surface waters,Microcystis aeruginosa and Chlamydomonas microsphaera,were exposed to 0,0.1,0.5,1.0,2.0,5.0,10.0,and 20.0 mg/L of oxytetracycline dihydrate(OXY) and sulfamethoxazole(SMZ) for 96 h to determine the effects of these antibiotics on the growth and surface morphology.Moreover,the photosynthetic activity and the contents of superoxide dismutase(SOD),malondialdehyde(MDA),and protein were measured to examine the biochemical characteristics of M.aeruginosa and C.microsphaera under OXY and SMZ stress.The effects of both antibiotics on the growth of both species were concentration-dependent and characterized by low-dose stimulation and high-dose inhibition.C.microsphaera was more sensitive to both antibiotics than M.aeruginosa was.The algal cell membranes of both species disintegrated after exposure to a high concentration of OXY.All of the physiological parameters measured in this study were relatively stable at low concentrations of OXY and SMZ.After exposure to high concentrations of OXY and SMZ,photosynthetic activity decreased significantly,whereas lipid peroxidation and the abundance of SOD,MDA,and protein increased significantly.Thus,low-dose antibiotics may increase algal blooms in eutrophic waters.     

PHYSIOLOGICAL INHIBITORY EFFECT OF OCS IN ARACHIDONIC ACID-RICH PARIETOCHLORIS INCISA （TREBOUXIOPHYCEAE, CHLOROPHYTA）

Parietochloris incisa is an arachidonic acid-rich snow green alga. The main physiological profiles, such as ash free dry weight (AFDW), chlorophyll, carotenoid, protein and total fatty acids (TFA), in this alga exposed to old culture supernatant (OCS) at the decline phase or its crude ethyl acetate extracts (CEAE) were investigated by using tubular photobioreactors of different diameters. Results showed that both OCS and CEAE had strong inhibitory effect on the above physiological parameters. The longer the culture was exposed to OCS and the more CEAE were added into the algal culture, the more the above physiological properties were inhibited. Arachidonic acid (AA), the dominant component of fatty acids in this alga, was also seriously inhibited with respect to total TFA, AFDW of cell mass, or culture volume, due to a prebable reduction of enzymes activities catalyzing chain elongation from C18:1ω9 to AA. These results incontestably evidenced that some CEAE dissolving substances existing in OCS, like auto-inhibitors, inhibited P. incisa growth through feedback. Hence, any efficient removal of auto-inhibitors from algal culture to decrease their bioactivity could be good for maximal production of desired products like AA.