Recover vigorous cells of Magnetospirillum magneticum AMB-1 by capillary magnetic separation

Cultivable magnetotactic bacteria (MTB) in laboratory can provide sufficient samples for molecular microbiological and magnetic studies. However, a cold-stored MTB strain, such as Magnetospirillum magneticum AMB-1, often loses its ability to synthesize magnetosomes and consequently fails to sense the external magnetic field. It is therefore important to quickly recover vigorous bacteria cells that highly capable of magnetosome producing. In this study, a modified capillary magnetic separation system was designed to recover a deteriorating strain of Magnetospirillum magneticum AMB-1 that long-term cold-stored in a refrigerator. The results show that all cells obtained after a 3-cycle treatment were vigorous and had the ability to produce magnetosomes. Moreover, the 3^rd-cycle recovered cells were able to form more magnetosome crystals. Compared with the colony formation method, this new method is time-saving, easily operated, and more efficient for recovering vigorous MTB cells.     

Characterization of Mediterranean Magnetotactic Bacteria

Magnetotactic bacteria are a diverse group of motile prokaryotes that are ubiquitous in aquatic habitats and cosmopolitan in distribution. In this study, we collected magnetotactic bacteria from the Mediterranean Sea. A remarkable diversity of morphotypes was observed, including multicellular types that seemed to differ from those previously found in North and South America. Another interesting organism was one with magnetosomes arranged in a six-stranded bundle which occupied one third of the cell width. The magnetosome bundle was evident even under optic microscopy. These cells were connected together and swam as a linear entire unit. Magnetosomes did not always align up to form a straight linear chain. A chain composed of rectangle magnetosomes bent at a posi- tion with an oval crystal. High resolution transmission electron microscopy analysis of the crystal at the pivotal position suggested uncompleted formation of the crystal. This is the first report of Mediterranean magnetotactic bacteria, which should be useful for studies of biogeochemical cycling and geohistory of the Mediterranean Sea.     

海相沉积岩磁化率的影响因素

磁化率作为最基础的环境磁学参数,在地层对比划分、环境气候研究中发挥着重要作用。因此,弄清磁化率的影响因素,对保证研究成果的可靠性具有关键意义。沉积岩(物)中磁性矿物的来源复杂,主要分为碎屑来源和自生成因两种。气候变化、海平面波动、碳酸盐产率、火山作用、陨石撞击等影响碎屑来源的磁性矿物,而磁性矿物还原作用、热液流体活动、趋磁细菌生产等因素影响自生成因的磁性矿物,这些因素综合决定了沉积岩的磁化率。      

Occurrence and microscopic analyses of multicellular magnetotactic prokaryotes from coastal sediments in the Yellow Sea

Multicellular magnetotactic prokaryotes(MMPs) are a group of aggregates composed of 7-45 gram-negative cells synthesizing intracellular magnetic crystals.Although they are thought to be globally distributed,MMPs have been observed only in marine environments in America and Europe.Most MMPs share a rosette-like morphology and biomineralize iron sulfide crystals.In the present study,abundant MMPs were observed,with a density of 26 ind./cm3,in the sediments of a coastal lagoon,Lake Yuehu,in the Yellow Sea.Optical microscopy showed that all of them were rosette shaped with a diameter of 5.5±0.8 mm.Transmission electron microscopy revealed that these MMPs were composed of 10-16 ovoid cells and flagellated peritrichously.High-resolution transmission electron microscopy and energy dispersive X-ray analysis indicated that they biomineralized bullet-shaped magnetite crystals in highly organized parallel chains within which the magnetosomes were oriented in the same direction.This is the first report of MMPs from Asia and demonstrates the ubiquitous distribution of MMPs.     

Metamorphosis of Magnetospirillum magneticum AMB-1 cells

Magnetospirillum magneticum strain AMB-1 belongs to the family of magnetotactic bacteria. It possesses a magnetosome chain aligning, with the assistance of cytoskeleton filaments MamK, along the long axis of the spiral cells. Most fresh M. magneticum AMB-1 cells exhibit spiral morphology. In addition, other cell shapes such as curved and spherical were also observed in this organism. Interestingly, the spherical cell shape increased steadily with prolonged incubation time. As the actin-like cytoskeleton protein MreB is involved in maintenance of cell shapes in rod-shaped bacteria such as Escherichia coli and Bacillus subtilis, the correlation between MreB protein levels and cell shape was investigated in this study. Immunoblotting analysis showed that the quantity of MreB decreased when the cell shape changed along with incubation time. As an internal control, the quantity of MamA was not obviously changed under the same conditions. Cell shape directs cell-wall synthesis during growth and division. MreB is required for maintaining the cell shape. Thus, MreB might play an essential role in maintaining the spiral shape of M. magneticum AMB-1 cells.     

Screening on DHA-producing Antarctic bacteria N- 6 and its cultural conditions

1 IntroductionPolyunsaturated fatty acids (PUFAs) have remained as a natural product due to thesynthetic difficulty of reproducing the methylene interrupted double bond sequence by in-dustrial chemistry. Their significance to animals and humans lies in th…     

Ecological Effects of Remote Smallsand Descending on Urban Area:Deduced from the "Mud - Snow" in Changchun City

IntroductionContinued sandstorms occurring in North Chinaeally this year have attracted close attention of manyscientists from different aspects. It is recorded thatserious sandstorms occurred eight times in China in1960' s, thirteen times in 1970, s, fou…     

Ecological Effects of Remote Smallsand Descending on Urban Area: Deduced from the "Mud- Snow" in Changchun City

A serious sandstorm occurred in North China from April 5 to April 7, 2000, while the remote smallsand decending took place in even larger area to the far northeast China. In Chanchun city, smallsand descending with snow provides the best samples to conduct a quantitative research on the possible impacts of city- ecology. In the 253km2 urban district area of Changchun city, smallsands accumulated up to more than 3000t on April 7, according to the calculation of four "mud- snow" samples. After naturally melting in room temperature, two samples are taken to do quarantine of "invasion bacteria". As a preliminary result, the individual amount of bacterium in 1 ml sample water is up to 8.9 × 104 or 1.05 × 105, and it is estimated that there are 640 million "invasion bacterium" individuals in one square meter. This study proves that other than the obvious environmental effects of sandstorm, the urban micro- ecosystem is very possible under threat by the "invasion bacterium" accompanied by remote descending smallsand.     

SCREENING FOR ANTIBACTERIAL AND ANTIFUNGAL ACTIVITIES IN SOME MARINE ALGAE FROM THE FUJIAN COAST OF CHINA WITH THREE DIFFERENT SOLVENTS

Three different solvents viz ethanol, acetone and methanol-toluene (3:1) were used to extract antibiotics from 23 species of marine algae belonging to the Chlomphyta, Phaeophyta and Rhodophyta. Their crude extracts were tested for antibacterial and antifungal activities. Among them, the ethanol extract showed the strongest activity against the bacteria and fungi tested. Four species of the Rhodophyta (Laurenc/a okamurai, Dasya scoparia, Grateloupia filicina and plocamium telfairiae ) showed a wide spectrum of antibacterial activity. Every solvent extract from the four species was active against all the bacteria tested. The test bacterium Pseudomonas solancearum and the fungus Penicilium citrinum were most sensitive to the extracts of marine algae. In general, the extracts of seaweeds inhibited bacteria more strongly than fungi and species of the Rhodophyta showed the greatest activity against the bacteria and fungi tested.     

Inhibitory activity of an extract from a marine bacterium Halomonas sp. HSB07 against the red-tide microalga Gymnodinium sp. （Pyrrophyta）

In recent years, red tides occurred frequently in coastal areas worldwide. Various methods based on the use of clay, copper sulfate, and bacteria have been successful in controlling red tides to some extent. As a new defensive agent, marine microorganisms are important sources of compounds with potent inhibitory bioactivities against red-tide microalgae, such as Gymnodinium sp. （Pyrrophyta）. In this study, we isolated a marine bacterium, HSB07, from seawater collected from Hongsha Bay, Sanya, South China Sea. Based on its 16S rRNA gene sequence and biochemical characteristics, the isolated strain HSB07 was identified as a member of the genus Halomonas. A crude ethyl acetate extract of strain HSB07 showed moderate inhibition activity against Gymnodinium sp. in a bioactive prescreening experiment. The extract was further separated into fractions A, B, and C by silica gel column chromatography. Fractions B and C showed strong inhibition activities against Gymnodinium. This is the first report of inhibitory activity of secondary metabolites of a Halomonas bacterium against a red-tide-causing microalga.     

Middle Holocene Organic Carbon and Biomarker Records from the South Yellow Sea: Relationship to the East Asian Monsoon

The East Asian monsoon system influences the sedimentation and transport of organic matter in East Asian marginal seas that is derived from both terrestrial and marine sources. In this study, we determined organic carbon (OC) isotope values, concentrations of marine biomarkers, and levels of OC and total nitrogen (TN) in core YSC-1 from the central South Yellow Sea (SYS). Our objectives were to trace the sources of OC and variations in palaeoproductivity since the middle Holocene, and their relationships with the East Asian monsoon system. The relative contributions of terrestrial versus marine organic matter in core sediments were estimated using a two-end-member mixing model of OC isotopes. Results show that marine organic matter has been the main sediment constituent since the middle Holocene. The variation of terrestrial organic carbon concentration (OCter) is similar to the EASM history. However, the variation of marine organic carbon concentration (OCmar) is opposite to that of the EASM curve, suggesting OCmar is distinctly influenced by terrestrial material input. Inputs of terrestrial nutrients into the SYS occur in the form of fluvial and aeolian dust, while concentrations of nutrients in surface water are derived mainly from bottom water via the Yellow Sea circulation system, which is controlled by the East Asian winter monsoon (EAWM). Variations in palaeoproductivity represented by marine organic matter and biomarker records are, in general, consistent with the recent EAWM intensity studies, thus, compared with EASM, EAWM may play the main role to control the marine productivity variations in the SYS.     

卵形鲳鲹结节病病原的分离与鉴定

从湛江港、阳江闸坡海水网箱养殖的患结节病卵形鲳鲹(Trachinotus ovatus)中分离一株细菌,回归感染结果证明该菌是引起卵形鲳鲹结节病的病原,对卵形鲳鲹半致死浓度为45 g-1。该菌株呈革兰阳性,好氧,具有弱抗酸性,菌体呈短杆状或细长分枝状,过氧化氢酶阳性,氧化酶阴性,还原硝酸盐,不水解酪素、黄嘌呤、酪氨酸、淀粉、明胶,以柠檬酸盐为唯一碳源生长。对其16S rDNA序列作Blast分析,构建系统进化树。结果表明,该菌株与诺卡氏菌属的菌株亲缘关系最近,且与鰤鱼诺卡氏菌(Nocardia seriolea JCM 3360T)的16S rDNA序列同源性高达99%。综合患病鱼的症状及病原菌形态、生理生化特性及16S rDNA序列的结果,该菌被鉴定为鰤鱼诺卡氏菌(N.seriolea)。     

红鳍笛鲷弧菌病病原的研究

从广东湛江特呈岛鱼排取患病红鳍笛鲷进行病原分离、纯化,得到一株病原菌Ls 1。该菌 为革兰阴性短杆菌,有运动性,菌落半透明,最适温度28～30℃,需盐生长,氧化酶反应阳性,发酵 葡萄糖不产气,不发酵肌醇,对弧菌抑制剂O/129敏感。经API ID32E细菌鉴定系统及弧菌科细 菌生化鉴定管鉴定,该菌为一株溶藻弧菌(Vibrioaginolyticus);其对红鳍笛鲷的半数致死量为6.32 ×105mL-1。药敏试验结果表明,该菌对阿莫西林、先锋V不敏感,对新生霉素、卡那霉素、多粘菌 素B、四环素中度敏感,而对庆大霉素、氟哌酸、氯霉素、红霉素、链霉素和复方新诺明等有较高的敏 感性。     

一株海水氨氧化细菌的系统发育分析

应用PCR技术扩增16S rRNA基因和amoA(ammonia monooxygenase subunit A)的基因片段,并测定其序列,对一株源于海水养殖水体的高效氨氧化细菌(ammonia-oxidizing bacteria,AOB)进行了系统发育分析。结果表明,经PCR扩增得到了1 098 bp的16S rRNA基因片段和491 bp的amoA基因片段,将其序列用NCBI-Blast软件在GenBank数据库中进行同源性检索后发现,该菌株的16S rRNA基因序列和amoA基因序列分别与亚硝化单胞菌Nitrosomonas sp.NS20的相对应基因片段相似性分别为98.4%和96.7%。在此基础上构建了系统发育树,表明用该菌株的16S rRNA基因片段和amoA基因片段构建的系统发育树均与亚硝化单胞菌属类聚在一起,结合该菌株形态和生理生化特性,鉴定该株氨氧化细菌属亚硝化单胞菌。     

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.     

Purification and characterization of 2-haloacid dehalogenase from marine bacterium Paracoccus sp. DEH99, isolated from marine sponge Hymeniacidon perlevis

2-haloacid dehalogenases constitute a group of dehalogenases which are capable of dehalogenating the halogenated organic compounds. So far, the 2-haloacid dehalogenases have been found in many bacteria, but not in Paracoccus genus. In the present study, one enzyme 2-haloacid dehalogenase(designated as Deh99), induced by DL-2-chloropropionate(DL-2-CPA), was purified from the marine bacterium Paracoccus sp. DEH99, isolated from marine sponge Hymeniacidon perlevis. The enzyme of Deh99 was purified to homogeneity by ammonium sulfate precipitation, ion exchange chromatography(Q-Sepharose HP), and Superdex 200 gel filtration chromatography. The molecular weight of Deh99 was estimated to be 25.0 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE), and 50.0 kDa natively by gel filtration chromatography. The enzyme of Deh99 stereospecifically dehalogenated L-2-CPA to produce D-lactate, with an apparent Michaelis-Menten constant(Km) value of 0.21 mmol L-1 for L-2-CPA. The optimal pH and temperature for Deh99 activity were 10.0 and 40℃, respectively. The enzyme of Deh99 acted on short-carbon-chain 2-haloacids, with the highest activity towards monochloroacetate. The activity of Deh99 was slightly affected by DTT and EDTA, but strongly inhibited by Cu2+ and Zn2+. The enzyme of Deh99 shows unique substrate specificity and inhibitor sensitivities compared to previously characterized 2-haloacid dehalogenases and is the reported one about purified 2-haloacid dehalogenase isolated from the bacteria of Paracoccus genus.     

一株拮抗芒果炭疽菌海洋细菌的鉴定和发酵培养基优化

从湛江近海海水和海泥分离的229株细菌中,选出对芒果炭疽菌(Colletotrichum gloeosporioides)具有较强拮抗作用的海洋细菌BSW03,抑菌圈直径为14.0 mm。根据菌体形态、生理生化特性和16s rDNA序列分析,BSW03鉴定为枯草芽孢杆菌(Bacillus subtilis)。从9种培养基中筛选出BDPB作为BSW03的发酵培养基,通过单因子碳源、氮源试验和L9(34)正交试验对发酵培养基成分进行优化。结果表明:发酵培养基最佳配方为乳糖20 g/L,牛肉膏15 g/L,K2HPO3.3H2O 0.75 g/L,MgSO4 0.75 g/L,pH自然。转速170 r/min、温度28℃的恒温摇床培养3 d后的无菌发酵液抑菌效果最佳,抑菌圈直径为27.3 mm。     

山东苍山三合地区磁异常特征及地质意义

通过对山东苍山三合地区高精度磁测磁异常的揭露,利用钻孔剖面对磁化强度的大小、磁性体的形态、磁性体的厚度进行反演和分析,认为研究区的磁异常是由该区辉绿岩磁性层引起的,并且辉绿岩层的上部相对较强磁性物质的富集对磁异常的影响更大;研究区辉绿岩层可能是普遍存在的;ZK02孔相对ZK01孔部位的辉绿岩层具有埋藏浅、磁性均匀、厚度变化小的特点。研究区磁异常的原因可能是成岩后局部断裂构造的活动,部分磁性块体边端部位挠起引起的。     

基于磁异常信息的鄂东北四方山地区沉积变质型锰矿找矿预测

为系统开展鄂东北地区锰矿找矿预测, 以四方山地区典型沉积变质型锰矿床为重点解剖对象, 结合高精度磁法测量异常信息, 查明磁异常与锰矿床(体)的空间对应关系。四方山地区锰矿床主要以层状或似层状赋存于新元古代红安岩群黄麦岭岩组(Pt₃h)中, 含锰岩系为一套含火山物质、泥质、粉砂质碎屑沉积岩夹碳酸盐岩, 属于陆源碎屑-碳酸盐岩系。含锰岩系因含少量磁铁矿等铁氧化物而多与1∶5万磁法测量中高强度磁异常套合, 但也有部分位于正负磁异常梯度带上。1∶5万磁法总梯度模量中锰矿化富集地段往往与中-低值区(尤其是线性梯度带)对应。基于矿床地质特征、磁法测量资料的二次开发及野外调查成果, 在四方山地区圈定了四个找矿靶区。其中四方山矿区北侧找矿靶区Ⅰ经TC160揭露一层锰矿体, 厚度2.53 m, Mn平均品位为14.63%;四方山矿区内找矿靶区Ⅱ北段经探槽TC140揭露两层锰矿体, 第一层厚1.63 m, Mn平均品位达24.57%;第二层厚0.98 m, Mn品位为16.73%;南段经探槽TC114揭露一层锰矿体, 厚度1.32 m, Mn平均品位为17.60%。团山沟矿区东南部找矿靶区Ⅳ经地质路线调查也发现了锰矿化体, 显示具有良好的找矿前景。