Importance of biofilm formation for corrosion inhibition of SAE 1018 steel by axenic aerobic biofilms

To investigate if corrosion inhibition by aerobic biofilms is a general phenomenon, carbon steel (SAE 1018) coupons were exposed to a complex liquid medium (Luria–Bertani) and seawater-mimicking medium (VNSS) containing fifteen different pure-culture bacterial suspensions representing seven genera. Compared to sterile controls, the mass loss in the presence of these bacteria (which are capable of developing a biofilm to various degrees) decreased by 2- to 15-fold. The extent of corrosion inhibition in LB medium depended on the nature of the biofilm: an increased proportion of live cells, observed with confocal scanning laser microscopy (CSLM) and image analysis, decreased corrosion. Corrosion inhibition in LB medium was greatest with Pseudomonas putida (good biofilm formation), while metal coupons exposed to Streptomyces lividans in LB medium (poor biofilm formation) corroded in a manner similar to the sterile controls. Pseudomonas mendocina KR1 reduced corrosion the most in VNSS. It appears that only a small layer of active, respiring cells is required to inhibit corrosion, and the corrosion inhibition observed is due to the attached biofilm. Received 09 December 1996/ Accepted in revised form 19 March 1997     

利用雨养泥炭沼泽及湖泊沉积物重建多氯联苯污染时空变化趋势

为了重建不同历史时期多氯联苯(PCBs)的沉降速率以研究其污染的时空变化趋势,本研究分析了PCBs在泥炭芯和湖泊沉积柱芯中浓度和沉降速率的变化规律,并评估了泥炭芯和湖泊沉积柱芯用于PCBs沉降变化研究的适用性.对采集柱芯进行定年分析发现,泥炭沼泽可以很好地记录PCBs历史沉降变化,并且泥炭中的PCBs没有出现降解情况,...     

Detection of heavy metals in bacterial biofilms and microbial flocs with the fluorescent complexing agent Newport Green

The complexing agent Newport Green fluoresces upon binding of nickel, zinc or cobalt. It was used to detect nickel or zinc in MOPS buffer, in gel-like matrices, and in natural biofilms and microbial flocs cultivated in the laboratory. The response curves for increasing nickel concentrations indicated an equimolar binding capacity of Newport Green for nickel in MOPS buffer, whereas zinc fluorescence reached saturation in the presence of a 10-fold excess of zinc ions relative to Newport Green molecules. The maximum fluorescence intensity as determined by luminometry was 8-fold and 4-fold above background for nickel and zinc, respectively. The response of Newport Green to either nickel or zinc in the presence of the other metal is consistent with a different binding affinity of Newport Green for the two metals. Zinc binds more strongly to the complexing agent than nickel but it leads to a weaker fluorescent signal which was detectable by luminometry but not by confocal laser scanning microscopy (CLSM). Newport Green was able to complex nickel in the presence of 1% gelatin or agarose as determined by CLSM and image processing. Its application to fully hydrated bacterial biofilms or microbial flocs revealed the presence of nickel outside of cells. The results suggest that in addition to cellular sorption, metals are bound extracellularly by extracellular polymeric substances in intact and undisturbed microbial aggregates. Journal of Industrial Microbiology & Biotechnology (2000) 24, 116–123. Received 11 June 1999/ Accepted in revised form 04 November 1999     

Bioremediation of Aroclor 1242 by a consortium culture in marine sediment microcosm

Plant terpenes have proven to be effective in stimulation of polychlorinated biphenyls (PCBs) biodegradation in soil systems. However, data on the application of plant terpenes in marine sediments contaminated with PCBs remains limited. The aim of this study was to ascertain the roles of a PCB degrading consortium and plant terpenes in stimulation of PCB biodegradation in marine sediments. The consortium culture 1-2Mix (strains 1-2M and 1-2T in commensalism), a utilizer of biphenyl and a natural substrate was enriched and isolated from marine sediments from the Busan coast, South Korea. PCB degradation by this culture was shown to be more effectively induced by tangerine peel extract than other known substrates (limonene, pinene, and cymene). Coastal sediment microcosms inoculated with 1-2Mix were set up to elucidate the effect of the consortium and plant terpenes on degradation of Aroclor 1242. After four weeks, the highest removal rates of PCBs, compared with the control (autoclaved sediment and no inoculation of 1-2Mix), were observed in order of the inducers tested; biphenyl (71.1%), tangerine peel extract (69.5%), surfactant (66.0%), and limonene (63.0%). Bioaugmentation effect was doubled in the presence of natural substrates such as tangerine peel extract and limonene, indicating effectiveness of these substrates in biostimulation. It was concluded that the tangerine peel extract could replace biphenyl as a feasible induction substrate for effective remediation of PCBs in the marine sediment.     

Polychlorinated biphenyls in vegetable soils from Changchun,Northeast China: Concentrations,distribution, sources and human health risks

To investigate the occurrences of polychlorinated biphenyls (PCBs) in suburban vegetable soils of Changchun area, Northeast China, 106 urban vegetable soil samples were collected from Changchun City, Nongan County, Dehui City, Yushu City, Jiutai City, Shuangyang District. We analyzed the concentrations, compositions and sources of 7 PCBs in top soils of Changchun area, and evaluated the non-carcinogenic and carcinogenic risk of PCBs pollution to exposed population. The total concentrations of 7 PCBs ranged from 1.31 to 148 ng/g dry weight (dw) with a mean value of 42.0 ng/g and dominated by Hepta-CBs and Penta-CBs. Principal component analysis (PCA) revealed that PCB pollution in soils of Changchun area mainly related to transportation, vehicle emissions, paints and other industries. Human health risk assessment showed that the cumulative non-carcinogenic and the cumulative carcinogenic risk in children and adults in the industrial land and residential land were acceptable, considering only 7 PCBs homologues were analyzed in this study, the actual risk could be higher.     

Isolation and Characterization of Acidic Peptides in Soy Sauce

A soy sauce sample was fractionated by gel filtration on a Sephadex G–15 column, then the fractions were subfractionated on the basis of acidity by ion exchange chromatography on a QAE-Sephadex A–25 column. The acidic subfractions with various acidities were further fractionated, using a preparative amino acid analyzer and by paper chromatography to separate the acidic peptide components.

Four dipeptides and sugar derivatives of ten dipeptides and two tripeptides were isolated and characterized as the major acidic peptides in soy sauce. However, it was difficult to anticipate any direct contribution of these peptides to the flavor construction in soy sauce on the basis of their contents and taste intensities.     

Ecotoxicological Risk Assessment of Polychlorinated Biphenyls (PCBs) in Bank Sediments from along the Yamuna River in Delhi,India

The River Yamuna originates from the Yamunotri glacier of the Himalayas and travels 22 km in the Delhi region. The river is used for various purposes in Delhi including drinking water supply. Twenty-eight polychlorinated biphenyls (PCBs) congeners were measured in bank sediments along the river, and their ecotoxicological risk was evaluated. Concentrations of ∑28PCBs varied from 0.20–21.16 ng g^?1 (dry wt.) with mean and median values of 6.63 ng g^?1 and 5.84 ng g^?1 (±0.69 ng g^?1), respectively. The concentration of 12 dl-PCBs concentrations varied from 0.04–2.86 ng g^?1 with a mean of 1.04 ± 0.11 ng g^?1, and their toxic equivalency ranged between <0.01–28.67 pg WHO-TEQ g^?1 with a mean of 10.77 ± 1.06 pg WHO-TEQ g^?1. CB-37, CB-44, CB-114, and CB-118 congeners were dominant among all PCBs congeners. The tri-PCBs (49%) were the main contributors to the PCB homolog followed by tetra-PCBs (35%), and penta-PCB (14%). Because there are no environmental guidelines in India for PCBs in river and marine sediments, concentrations of PCBs and their toxic equivalents were compared in a screening-level assessment with established freshwater sediment quality guidelines and found lower than those guideline values, which suggests no adverse ecotoxicological effect.     

用免疫荧光标记对人组织中肥大细胞亚型的分选与形态观察

方法:利用中性蛋白酶成分、特征性酶抗体的免疫荧光染色和流式细胞仪确定分选肥大细胞亚型,以激光扫描共聚焦显微镜显示肥大细胞内分泌颗粒。结果:三种免疫表型被确定:肥大细胞的类胰蛋白酶阳性(MCT)、类糜蛋白酶阴性;类糜蛋白酶阳性(MCC)、类胰蛋白酶阴性和类胰蛋白酶阳性、类糜蛋白酶阳性(MCTC)。肥大细胞内分泌颗粒分散或聚集存在,分泌颗粒突起分泌或以分散的方式释放。分泌颗粒大范围释放后,肥大细胞的形态结构发生了改变。结论:利用肥大细胞的特征性酶抗体、免疫荧光标记和流式细胞仪可将人组织中的肥大细胞分选纯化为三种亚型;以共聚焦显微镜显示肥大细胞含有丰富的分泌颗粒,它说明肥大细胞具备了为人体I型变态反应提供快速反应的物质基础。     

Potential for phytoremediation of polychlorinated biphenyl-(PCB-)contaminated soil

Weathered soils contaminated with commercial-grade Aroclor 1260 from three sites in Canada were used to investigate the polychlorinated biphenyl (PCB) phytoextraction potential of nine plant species (Festuca arundinacea, Glycine max, Medicago sativa, Phalaris arundinacea, Lolium multiflorum, Carex normalis, and three varieties of Cucurbita pepo ssp. pepo) under controlled greenhouse conditions. The soils used varied in PCB concentration (90-4200 microg/g) and total organic content (0.06-2.02%). Greenhouse experiments controlled for PCB volatilization through the use of a vented enclosure and by isolating the contaminated soils with parafilm. After 8 wks, PCB concentrations of 47-6700 microg/g were observed in root tissues. Although PCB concentrations in shoot tissues were lower (< 1-470 microg/g), the absolute amounts of PCBs observed in shoot tissue were significant (1.7-290 microg) once shoot biomass was accounted for. Congener signatures indicated that tetra- to hexa-chlorobiphenyls contributed the largest proportions to shoot tissues, but hepta-to nona-chorobiphenyls were also present in measurable amounts. Overall, the results indicate that varieties of C. pepo were more effective at extracting PCBs from soil than other plants screened The evidence suggests that this was mainly due to root uptake of PCBs and tranlocation to the shoots, rather than volatilization of PCBs from soil. All plants screened showed signs of stress in the most highly contaminated soil (4200 microg/g), but not in the two lower PCB contaminated soils (250 and 90 microg/g, respectively). No detectable decreases in soil PCB concentrations were observed in these short-term greenhouse experiments, but the results suggest that this may be achievable through multiple plantings.     

Colonization of barley (Hordeum vulgare) with Salmonella enterica and Listeria spp

Colonization of barley plants by the food-borne pathogens Salmonella enterica serovar typhimurium and three Listeria spp. (L. monocytogenes, L. ivanovii, L. innocua) was investigated in a monoxenic system. Herbaspirillum sp. N3 was used as a positive control and Escherichia coli HB101 as a negative control for endophytic root colonization. Colonization of the plants was tested 1-4 weeks after inoculation by determination of CFU, specific PCR assays and fluorescence in situ hybridization (FISH) with fluorescently labelled oligonucleotide probes in combination with confocal laser scanning microscopy (CLSM). Both S. enterica strains were found as endophytic colonizers of barley roots and reached up to 2.3 x 10(6) CFU per g root fresh weight after surface sterilization. The three Listeria strains had 10-fold fewer cell numbers after surface sterilization on the roots and therefore were similar to the results of nonendophytic colonizers, such as E. coli HB101. The FISH/CSLM approach demonstrated not only high-density colonization of the root hairs and the root surface by S. enterica but also a spreading to subjacent rhizodermis layers and the inner root cortex. By contrast, the inoculated Listeria spp. colonized the root hair zone but did not colonize other parts of the root surface. Endophytic colonization of Listeria spp. was not observed. Finally, a systemic spreading of S. enterica to the plant shoot (stems and leaves) was demonstrated using a specific PCR analysis and plate count technique.     

F肌动蛋白作为胞间连丝组分的结构与生理学证据

以蒜 (AlliumsativumL .)瓣鞘外表皮为材料 ,利用荧光特异探针与共焦镜检术 ,结合透射电镜与免疫金标记对表皮细胞间胞间联络的性质、结构进行了系统观察。结果表明 ,加厚壁上的通道是由狭长的管状胞质和初生纹孔场上成束的胞间连丝衔接组成 ,前者实为原生质体的一部分。单个胞间连丝的孔径为 6 0～ 70nm ,属正常胞间连丝范围 ,它们乃相邻细胞间共质联系的所在。荧光探针TRITC_Phalloidin (TRITC_Ph)标记的结果显示 ,整个通道上呈现红色荧光的纤索在接近初生纹孔场处明显变窄 ,与超微结构观察中所见的结构特点相吻合 ,共焦镜下观察到的初生壁上的荧光亮斑乃初生纹孔场中成束胞间连丝被标记的反映 ,从而有效地证实了F肌动蛋白在常态胞间连丝上的存在。免疫金标记实验显示在管状胞质中和胞间连丝上有金颗粒分布 ,这一结果为证实荧光标记物具肌动蛋白性质提供了有说服力的补充。     

F肌动蛋白作为胞间连丝组分的结构与生理学证据

以蒜(Allium sativum L.)瓣鞘外表皮为材料,利用荧光特异探针与共焦镜检术,结合透射电镜与免疫金标记对表皮细胞间胞间联络的性质、结构进行了系统观察.结果表明,加厚壁上的通道是由狭长的管状胞质和初生纹孔场上成束的胞间连丝衔接组成,前者实为原生质体的一部分.单个胞间连丝的孔径为60～70 nm,属正常胞间连丝范围,它们乃相邻细胞间共质联系的所在.荧光探针TRITC-Phalloidin (TRITC-Ph)标记的结果显示,整个通道上呈现红色荧光的纤索在接近初生纹孔场处明显变窄,与超微结构观察中所见的结构特点相吻合,共焦镜下观察到的初生壁上的荧光亮斑乃初生纹孔场中成束胞间连丝被标记的反映,从而有效地证实了F肌动蛋白在常态胞间连丝上的存在.免疫金标记实验显示在管状胞质中和胞间连丝上有金颗粒分布,这一结果为证实荧光标记物具肌动蛋白性质提供了有说服力的补充.     

利用激光共聚焦扫描显微镜对溃疡病菌侵染柑橘叶片的实时观察

柑橘溃疡病对柑橘产业造成了巨大损失,而研究柑橘与溃疡病菌的互作关系以及柑橘的感病和抗病性均需要观察溃疡病菌在柑橘寄主中的侵染和定殖过程。激光共聚焦扫描显微镜不仅可以观察活细胞,活组织的动态代谢过程,而且可以获得三维图像,对于病原菌在柑橘植物组织内的繁殖和致病机制研究具有重要意义。但是,选择适宜的植物材料和制片方法对激光共聚焦扫描显微镜的观察效果影响很大。本文对激光共聚焦扫描显微镜所观察的材料在其处理和观察方法上加以改进,获得了质量更好的图片和实验结果,也使得实验更为方便快捷。激光共聚焦扫描显微观察还在瞬时表达分析中得到应用,提高了柑橘瞬时表达分析的效果。通过将切片和压片相结合观察到溃疡病菌在不同时间点对柑橘叶片的侵染情况,而通过3D建模能观察到柑橘叶片不同组织层面中的病菌数量和病菌位置,为研究溃疡病菌在叶片中的定殖方式和入侵数量提供了前期基础。     

Body musculature of Beauchampiella eudactylota (Gosse, 1886) (Rotifera: Euchlanidae) with additional new data on its trophi and overall morphology

This study presents the results of confocal laser scanning microscopy and fluorescence‐labelled phalloidin used to visualize the system of body musculature in Beauchampiella eudactylota. Moreover, the poorly known trophi of B. eudactylota are described based on scanning electron microscopy. In total, four paired longitudinal muscles (musculi longitudinales I–IV) and three circular muscles (musculi circulares I–III) were identified. Among these are the musculus longitudinalis ventralis, the musculus longitudinalis dorsalis and the musculus circumpedalis as documented in previous studies for other rotifer species. Compared to other species, B. eudactylota is characterized by the low number of lateral longitudinal muscles and the absence of some longitudinal muscles (musculi longitudinales capitum) and circular muscles (corona sphincter, musculus pars coronalis). Moreover, scanning electron microscopic data on the trophi of B. eudactylota reveal a number of striking similarities to the trophi in some species of Epiphanidae. This suggests that either (1) these similarities represent plesiomorphic characters present both in Epiphanidae and B. eudactylota or (2) they are synapomorphic features of B. eudactylota and some species of Epiphanidae, which would question the monophyly of Euchlanidae.     

IPL作用下鼠皮形态改变的显微观察与初步分析

观察光子嫩肤(IPL)后鼠皮的形态学随时间的改变情况,如:表皮层、真皮层的增厚或变薄的情况。探讨IPL作用下,皮肤的表皮、真皮厚度变化与IPL在特定波长的能量密度的关系。以小白鼠作为研究对象,先去毛,用IPL在一定波长不同的能量密度照射活体小白鼠皮肤,分别在照射前、照射后,以及之后的1天,3天,7天应用激光共聚焦显微镜观察皮肤内部的结构,并对其不同的能量密度与照射前相比进行分析,讨论了真皮胶原蛋白在组织的修复过程的作用及其中的关键因素。     

利用激光扫描共聚焦显微术对同源四倍体水稻胚囊形成与发育的进一步观察

应用改进的整体染色透明激光扫描共聚焦显微术(WCLSM),对同源四倍体水稻PDER-2B-4x胚囊的形成与发育过程进行观察。发现其胚囊的形成发育过程与二倍体的一致,可以清楚地划分为8个发育时期,即孢原细胞形成期、大孢子母细胞形成期、大孢子母细胞减数分裂期、功能大孢子形成期、单核胚囊形成期、胚囊有丝分裂期、八核胚囊发育期和成熟胚囊期。除正常发育的过程外,大孢子发育的各个过程均出现一些异常现象,包括:细胞退化、核位置异常、核数目异常和细胞分化异常等。这些异常可能最终导致多种结构异常成熟胚囊的形成。     

Specific lectins map the distribution of fibronectin and beta 1-integrin on living MDCK cells

The expression and dynamics of bound fibronectin and the sialylated integral membrane protein, beta 1-integrin, were analyzed on the apical membrane of living MDCK cells. Fibronectin was identified by its specific binding of fluorescent peanut agglutinin and sialylated beta 1-integrin by its binding of Sambucus nigra agglutinin. Confocal epifluorescence microscopy and laser scanning cytometry determined the distribution and abundance of binding sites of the two fluorescently labeled lectins. Both fibronectin and beta 1-integrin were restricted to specific regions uniformly distributed over the entire apical surface. Apical-surface fibronectin binding varied much more between cells than did the expression of beta 1-integrin. Sialylated beta 1-integrin colocalized >92% with membrane microplicae while fibronectin was unrelated to these surface structures. This lack of colocalization of the proteins was confirmed by double-labeling experiments. From the maturation dependence of the fibronectin-binding capacity and the differences in protein turnover times, it was evident that fibronectin did not bind to sialylated beta 1-integrin. Furthermore, desialylation of beta 1-integrin uncovered additional fibronectin receptors on the apical membrane. We conclude that these lectins permit tracking of two membrane-associated glycoproteins in living cells and that fibronectin binds only to desialylated beta 1-integrin on MDCK cells.     

Bacillus cereus B-02对Botrytis cinerea拮抗机理的研究

采用电子显微镜(扫描、透射)和激光扫描共聚焦显微镜,从细胞形态学和生理学水平上研究蜡样芽孢杆菌Bacillus cereus B-02过滤液对灰葡萄孢菌Botrytis cinerea的拮抗机理。结果表明,处理菌丝表面形态受到严重破坏,发生强烈变形;菌丝细胞核、线粒体和细胞壁等亚细胞结构发生了明显改变,细胞内出现大量无膜透明内含物,并产生较大液泡。此外,处理菌丝DNA、线粒体膜电位和活性氧荧光强度均低于对照组,且差异极显著;说明B-02菌株对病原真菌菌丝细胞DNA的合成、线粒体膜电位和活性氧水平有重要影响。