具有多种胞外酶的对虾肠道黏附菌的分离和鉴定

用对虾饲料培养基从健康凡纳滨对虾肠道分离出500株黏附细菌,以产淀粉酶、脂肪酶和蛋白酶能力为指标,筛选出产该3种消化酶的细菌90株,占总菌株的18%.对其中生长较快的69株进行16S rDNA 基因测序,确定其分类地位.结果显示,69株菌分别属于不动杆菌属(Acinetobacter)、芽孢杆菌属(Bacillus)、葡萄球菌属(Staphylococcus)、假交替单胞菌属(Pseudoalteromonas)、气单胞菌属(Aeromonas)、嗜盐单胞菌属(Halomonas)、利斯顿氏菌属(Listonella)、莫拉氏菌属(Moraxella)等,其中数量最多是芽胞杆菌属,占鉴定细菌总数的53.62%,数量最少是气单胞菌属和嗜盐单胞菌属,均占鉴定细菌总数的2.90%.表明对虾肠道黏附菌群中具有较多能分泌多种消化酶的细菌,可进一步开发为促进对虾消化功能的益生菌     

拮抗香蕉枯萎病菌海洋细菌的筛选和鉴定

从93份中国南部海域近海海泥样品中分离出1800多株海洋细菌,以对峙培养法,测定海洋细菌对香蕉枯萎菌的拮抗作用,再以琼脂划线培养法(agar streak)测定菌株的抑菌活性,结果表明:约55.6%的菌株对香蕉枯萎菌有一定的拮抗作用,其中58株菌株抑制香蕉枯萎菌生长所形成的抑菌带宽度大于10.0mm,抑菌作用较好,复筛后其中13个菌株形成的抑菌带宽在15.0mm以上。经反复转接和对峙培养,筛选出拮抗效果好、抑菌活性稳定的菌株TC-1,初步鉴定为芽孢杆菌属的细菌,TC-1对14种病原微生物都有较好的抑菌效果。     

Characterization and in-vivo evaluation of potential probiotics of the bacterial flora within the water column of a healthy shrimp larviculture system

A thorough understanding of the normal bacterial flora associated with shrimp larviculture systems contributes to probiotic screening and disease control.The bacterial community of the water column over a commercial Litopenaeus vannamei larval rearing run was characterized with both culture-dependent and culture-independent methods.A total of 27 phylotypes at the species level were isolated and identified based on 16 S rDNA sequence analysis.Denaturing gradient gel electrophoresis(DGGE) analysis of the V3–V5 region of 16 S rRNA genes showed a dynamic bacterial community with major changes occurred from stages zoea to mysis during the rearing run.The sequences retrieved were affiliated to four phyla,Proteobacteria,Actinobacteria,Bacteroidetes,and Firmicutes,with the family Rhodobacteraceae being the most frequently recovered one.Subsequently,13 representative strains conferred higher larval survival than the control when evaluated in the in-vivo experiments;in particular,three candidates,assigned to Phaeobacter sp.,Arthrobacter sp.,and Microbacterium sp.,significantly improved larval survival( P 0.05).Therefore,the healthy shrimp larviculture system harbored a diverse and favorable bacterial flora,which contribute to larval development and are of great importance in exploiting novel probiotics.     

A bacterial pathogen infecting gametophytes of Saccharina japonica (Laminariales, Phaeophyceae)

A newly identified bacterial disease of kelp(Saccharina japonica) gametophytes was found in clone cultures.It is characterized by swollen gametophyte cells in the early period of infection followed by filamentous fading.An alginolytic marine bacterium referred to as A-1 was isolated from the diseased gametophytes.On the basis of 16S rDNA sequencing and morphological,physiological and biochemical characteristics,the bacterium was identified as a strain of the genus Alteromonas.By testing Koch’s postulates,Alteromonas sp.A-1 was further confirmed as the pathogen.The infection process was also investigated using both scanning electron and light microscopy.     

钻孔体应变仪高采样对比观测实验研究

为分析提高应变观测采样率及拓展观测频带对低频观测的影响,在张家口地震台、怀来地震台选择TJ-2型体应变仪进行台站对比观测实验,并在时域和频域分别对实验数据进行分析,总结体应变与测震观测频带的特性.     

一株分离自红树林沙雷氏菌的鉴定及杀虫活性测定

【目的】为开发用于害虫生物防治的细菌杀虫剂,对分离自红树林的一株沙雷氏菌进行鉴定并初步测试其杀虫活性。【方法】采用形态特征比较、细菌理化特性测定和基于16S rDNA序列构建系统发育树对从湛江红树林根际分离的菌株ZJ9进行综合鉴定,采用喂毒法和注射法进行杀虫活性测定。【结果】该菌为黏质沙雷氏菌Serratia marcescens,对草地贪夜蛾3龄幼虫7d校正死亡率为80.21%,且与苏云金芽孢杆菌和隐地杆菌无显著差异,菌体及菌液对大蜡螟老熟幼虫血腔注射24 h死亡率均达100%。【结论】菌株ZJ9为黏质沙雷氏菌,对草地贪夜蛾具有很好杀虫活性,可用作害虫的生物防治资源。     

黄喉拟水龟体表溃疡病原菌SG_(24)的分类鉴定

从患溃疡的黄喉拟水龟病灶中分离到一株病原菌SG24。对菌株SG24进行了常规生理生化测定和ATBExpression半自动细菌鉴定仪鉴定,并测定16S rRNA序列,分析其与相关细菌相应序列的同源性,构建了系统进化树。普通细菌学方法结果显示菌株SG24为黏质沙雷氏菌(Serratia marcescens)。以沙雷氏菌属的16S rRNA基因序列设计一对引物进行PCR扩增,获得了菌株SG24大小约950 bp的16S rRNA部分基因片段,测序结果显示菌株SG24与黏质沙雷氏菌同类,与已登录的黏质沙雷氏菌(S.marcescensDQ207558)的16S rRNA同源性大于99%。综合以上分类鉴定结果,确定菌株SG24属于沙雷氏菌属的黏质沙雷氏菌。药物敏感试验结果表明:菌株SG24对链霉素、庆大霉素、壮观霉素、强力霉素、卡那霉素、阿米卡星、诺氟沙星、氧氟沙星和复方新诺明敏感。     

Bioflocculant Produced by Klebsiella sp. MYC and Its Appli-cation in the Treatment of Oil-field Produced Water

1IntroductionWater co-produced with oil and gas recovery istermed oil-field produced water.This water is consid-ered to be of the largest volume in the waste streamfromoil and gas production processes.The volume ofproduced water can be more than10ti mes t…     

Impact of redox-stratification on the diversity and distribution of bacterial communities in sandy reef sediments in a microcosm

Relationships between microbial communities and geochemical environments are important in marine microbial ecology and biogeochemistry. Although biogeochemical redox stratification has been well documented in marine sediments, its impact on microbial communities remains largely unknown. In this study, we applied denaturing gradient gel electrophoresis (DGGE) and clone library construction to investigate the diversity and stratification of bacterial communities in redox-stratified sandy reef sediments in a microcosm. A total of 88 Operational Taxonomic Units (OTU) were identified from 16S rRNA clone libraries constructed from sandy reef sediments in a laboratory microcosm. They were members of nine phyla and three candidate divisions, including Proteobacteria (Alpha-, Beta-, Gamma-, Delta-, and Epsilonproteobacteria), Actinobacteria, Acidobacteria, Bacteroidetes, Chloroflexi, Cyanobacteria, Firmicutes, Verrucomicrobia, Spirochaetes, and the candidate divisions WS3, SO31 and AO19. The vast majority of these phylotypes are related to clone sequences from other marine sediments, but OTUs of Epsilonproteobacteria and WS3 are reported for the first time from permeable marine sediments. Several other OTUs are potential new bacterial phylotypes because of their low similarity with reference sequences. Results from the 16S rRNA, gene clone sequence analyses suggested that bacterial communities exhibit clear stratification across large redox gradients in these sediments, with the highest diversity found in the anoxic layer (15–25 mm) and the least diversity in the suboxic layer (3–5 mm). Analysis of the nosZ, and amoA gene libraries also indicated the stratification of denitrifiers and nitrifiers, with their highest diversity being in the anoxic and oxic sediment layers, respectively. These results indicated that redox-stratification can affect the distribution of bacterial communities in sandy reef sediments.     

Antifouling activities of marine bacteria associated with sponge (Sigmadocia sp.)

The present study aimed at assessing the antifouling activity of bacteria associated with marine sponges. A total of eight bacterial strains were isolated from the surface of sponge Sigmadocia sp., of them, SS02, SS05 and SS06 showed inhibitory activity against biofilm-forming bacteria. The extracts of these 3 strains considerably affected the extracellular polymeric substance producing ability and adhesion of biofilm-forming bacterial strains. In addition to disc diffusion assay, microalgal settlement assay was carried out with the extracts mixed with polyurethane wood polish and coated onto stainless steel coupons. The extract of strain SS05 showed strong microalgal settlement inhibitory activity. Strain SS05 was identified as Bacillus cereus based on its 16S rRNA gene. Metabolites of the bacterial strains associated with marine invertebrates promise to be developed into environment-friendly antifouling agents.     

Morphological and genetic comparison of two strains of a Prorocentrum species isolated from Zhejiang coastal water of China and Masan Bay of Korea

In this paper, we examined the detailed morphology of two strains of Prorocentrum isolated from the coastal waters of Zhejiang (Wenling area), China, and Masan Bay of Korea. A taxonomic comparison was made among strains on the basis of morphological and molecular data. The cellular dimensions of the Chinese Wenling strain (LAMB090508) and Korean strain (PDKS0206) were similar and the cells of both strains were of asymmetric and elongated shape. The posterior end of most cells was rounded. Megacytic zones of aged cells were broader with dense tiny knobs. The roundish nucleus was located in the posterior part of the cell. A few irregular shaped chloroplasts were distributed within the cell. The nucleotide similarity of the two strains, determined from the 5.8S rDNA-ITS sequences, was 99.83%. The comparative results of morphology and molecular analysis suggest that both strains isolated from China and Korea were the identical species, Prorocentrum donghaiense Lu.     

Bioflocculant Produced by Klebsiella sp. MYC and Its Application in the li＇eatment of Oil-field Produced Water

Seventy-nine strains of bioflocculant-producing bacteria were isolated from 3 activated sludge samples. Among them, strain MYC was found to have the highest and stable flocculating rate for both kaolin clay suspension and oil-field produced water. The bacterial strain was identified as Klebsiella sp. MYC according to its morphological and biochemical characteristics and 16SrDNA sequence. The optimal medium for bioflocculant production by this bacterial strain was composed of cane sugar 20 g L^-1, KH2PO4 2 g L^-1, K2HPO4 5 g L^- 1, （ NH4）2SO4 0.2 g L^-1, urea 0.5 g L^- 1 and yeast extract 0.5 g L^- 1, the initial pH being 5.5. When the suspension of kaolin clay was treated with0.5% of Klebsiella sp. MYC culture broth, the flocculating rate reached more than 90.0% in the presence of 500mg L^-1 CaCI2, while the flocculating rate for oil-field produced water was near 80.0% in a pH range of 7.0 - 9.0 with the separation of oil and suspended particles from the oil-field produced water under similar conditions. The environment-friendly nature of the bioflocculant and high flocculating rate of the strain make the bioflocculant produced by Klebsiella sp. MYC an attractive bioflocculant in oil-field produced water treatment.     

Isolation and characterization of a fucoidan-degrading bacterium from Laminaria japonica

Fucoidan, a polysaccharide containing abundant fucose and sulfate ester group, was prepared from Laminaria japonica. In order to obtain fucoidan-degrading enzyme, bacteria capable of degrading fucoidan were screened from kelp. A bacterial strain named RC2-3 was obtained, which degraded fucoidan by the maximum extent of 54% ± 1.3%, the highest among all bacterial isolates. High-performance size exclusion chromatography(HPSEC) showed that the molecular weight of fucoidan was gradually reduced by RC2-3 with culturing time, suggesting the production of fucoidan-degrading enzyme by RC2-3. Phylogenetic analysis of partial 16S ribosomal RNA gene(16S rDNA) sequence showed that RC2-3 belonged to the family Flavobacteriaceae. However, it showed different physiological and biochemical characteristics from the known Flavobacteriaceae members producing fucoidan-degrading enzyme, thus RC2-3 was proposed to be a new member of this family.     

Preliminary study on PAH degradation by bacteria from contaminated sediments in Xiamen Western Sea,Fujian, China

In order to estimate the biodegradation of three polycyclic aromatic hydrocarbons (PAHs) compounds, bacterial strains were isolated from marine sediments in three heavily contaminated sites (Yuandang Lake, Dongdu Port and Aquacultural zones in Maluan Bay) in Xiamen Western Sea. The results show three bacterial strains, which used pyrene as the sole carbon source, were identified as strains of Aureobacterium sp.,Arthrobacter sp., Rhodococcus sp. The PAH-degrading bacteria isolated had a strong ability to degrade phenanhrene, fluoranthene and pyrene at different degradation rates. The highest degradation rate was observed when three PAH compounds were mixed with an individual strain in the medium. The three PAHs were degraded after one week with a degradation rate of 89.94 % for phenanthrene and 93.4 % for both of fluoranthene and pyrene.In addition, after 25 days of incubation, the degradation rate was 99.98 % for phenanthrene and 99.97 % for both of fluoranthene and pyrene. Optical density was measured to estimate bacterial growth during the degradation of PAHs. Highest levels of bacterial growth were observed with a three PAH mixture in the culture, suggesting that the concentration of PAHs influenced bacterial growth and the highest levels of degradation for most series were detected after one week of incubation.     

Denitrification potential evaluation of a newly indigenous aerobic denitrifier isolated from largemouth bass Micropterus salmoides culture pond

This work evaluates the application potential of a new indigenous aerobic denitrifi er, strain Pseudomonas CW-2, isolated from a largemouth bass culture pond. The rate of ammonium-N removal by strain CW-2 was approximately 97% at a DO concentration of 5.2 mg/L. Furthermore, when nitrate and ammonia coexisted, the strain gave priority to assimilating ammonia, and thereafter to denitrifi cation. Under optimal cultivation conditions, citrate and acetate were the carbon resources, C/N was 8, dissolved oxygen was 5.2 mg/L, and pH was 7; the removal rate of ammonium reached nearly 90%. The changing patterns of different bacteria in strain CW-2-treated and the control pond water were also compared. Lower levels of ammonia, nitrite, and phosphates were observed in the treated water as compared with the controls. Meanwhile, phylum-level distributions of the bacterial OTUs revealed that P roteobacteria, Bacteroidetes, Planctomycetes, and N itrospirae continuously changed their relative abundances in relation to carbon and the addition of strain CW-2; this finding implies that the conventional denitrifi cation process was weakened under the ef fects of carbon or the presence of strain CW-2. We propose that strain CW-2 is a promising organism for the removal of ammonium in intensive fish culture systems, according to our evaluations of its denitrifi cation performance.     

Microalgae cultivation in a tubular bioreactor and utilization of their cells

In this study on the possiblities of microalgae technology as an option for CO₂ mitigation, many microalgae were isolated from seawater. Some species of the isolates,Chlamydomonas sp. strain YA-SH-1, which accumulates starch in cells under light and ferment ethanol in dark and anaerobic condition, was grown outdoors by using 50-L tubular bioreactors in batch cultivation and harvested. Using these cells, the performance of ethanol production was examined quantitatively in a 0.5-L scale fermentor. Another species,Tetraselmis sp. strain Tt-1, was cultivated in a semi-batch manner by a similar type of tubular bioreactor indoors and examined for its utilization. Tests showed these cells could be used as partial substitute for wood and kenaf pulp for processing into paper. With the idea of making microalgae produce cellulose by genetic engineering in their minds, the authors studied the structure of bacterial cellulose synthase genes and the low temperature-induced, reversible flocculation in a thermophilic blue green alga (Cyanobacterium),Synechocystis vulcanus in order to examine the feasibility of using these genes as gene source and the cynanobacterium as host.

     

Intraspecific crossings ofUndaria pinnatifida (Harv.) Sur.—A possible time-saving way of strain selection

Unicellular gametophytes ofUndaria pinnatifida (Harv.) Sur. were isolated in Qingdao, P. R. China in April 1993 and in Tokushima, southern Japan in March 1995. Different intraspecific crossings by using unicellular male and female gametophytes were successfully undertaken in Sept. of 1995 in Qingdao. Sporophytes were transplanted to two different locations for open sea cultivation. One was at Zhanqiao (ZQ) Bay where the water current was slower than that of another location—Taipingjiao(TPJ). A total of 218 adult sporophytes were harvested on January 12 and 18 from TPJ and ZQ respectively. For eacn combination, 10 sporophytes were cultivated. Analysis of the morphological characteristics of adult sporophytes indicated that the longest length between two bases of the serration of pinnate blades (W2) is a morphological characteristic that can be transferred from the parent plant to the next generations regardless of environmental variations. There was evidence that W2 was apparently determined by sex-linked factors, i.e., by male parental gametophyte. Sporophytes from certain crossing combinations showed more vigorous growth than those from other crossing combinations. It is therefore possible to select gametophyte strains which can be used as parental gametophytes for the seedling production of sporophytes with more vigorous growth within shorter cultivation period. The morphology of hybrids from a Qingdao strain and a Tokushima cultivated strain resembled that of both parental plants in frond features (wrinkled or smooth) and W2. Sporophyll formation also varied with strains. The fact that adult sporophytes resulting from the same crossing combinations have identical morphological characteristics under the same environmental conditions indicates the possibility of a new way to select strains which are expected to be ideal for commerical production by purposely selecting, propagating, and seeding unicellular gametophytes for sporeling production through freeliving techniques of gametophytes. Project 39400102 supported by NSFC, also supported by National Climbing Plan B (PD-B642); and Bioengineering Center, SSTC.     

Isolation and characterization of AHL-degrading bacteria from fish and pond sediment

Quorum sensing(QS)disruption is considered as a potential alternative strategy to combat bacterial diseases in aquaculture.In this study,we isolated and identified bacteria degrading QS molecules from pond sediment and fish intestine.A total of 132 strains were obtained in the enrichment culture,of which two strains were identified as Enterobacter sp.f003 and Staphylococcus sp.sw120,being isolated from the fish intestine and pond sediment,respectively.We found that strains f003 and sw120 could degrade acyl-homoserine lactones(AHLs)and cause no hemolysis of sheep red blood cells.The AHL lactonase(aiiA)homologous gene in the two strains was detected in PCR amplification and the high-degrading activity to N-hexanoyl-L-homoserine lactone(C6-HSL)and AHLs secreted from pathogenic Aeromonas hydrophila was assessed.Meanwhile,the artificial infection of cyprinid Carassius auratus gibelio with intraperitoneal injection showed that the two strains were avirulent.Therefore,the obtained indigenous bacteria are candidate probiotics against pathogenic A.hydrophila in aquaculture.     

潮汐线应变观测的地表平面应变状态

提出了由潮汐线应变的组合观测资料获取地表平面应变状态和对潮汐剪切应变进行数值模拟的方法。计算了我国部分潮汐应变观测点的理论应变状态和观测应变状态，并对结果进行了比较分析，为深入研究地震前兆的物理力学机制和观测台网的优化提供理论依据     

叠加变形有限应变分解及其应用探讨

本文提出了二维不同类型叠加变形的有限应变分解方法。只要测定有限应变及能代表各期主应变面的面理或剪切指向,就可以计算出各期构造应变。