Evaluation of epifluorescence image analysis of biofilm growth on stainless steel surfaces

Biofilm growth of Bacillus subtilis, Pseudomonas fragi, Pediococcus inopinatus and Listeria monocytogenes was studied on stainless steel surfaces at room and low temperatures to evaluate the results of traditional hygiene measures. The results were compared with those of image analysis of stainless steel surfaces in an epifluorescence microscope. Statistical analyses were carried out to determine the variations between the conventional cultivation swab method, the glycocalyx amount obtained using swabbing, and the values of the areas of the biofilm, slime and cells. As a general rule, old biofilms showed total counts at approximately the same levels as the young biofilm. The results showed that temperature affected the results for all strains except B. subtilis. The strains of Pe. inopinatus and Ps. fragi showed increased attachment at 6°C and L. monocytogenes at 25°C. The biofilm slime was more easily detached than the cells. The results indicated that the traditional swab method is not reliable for the measurement of biofilm formation on surfaces.     

A switch-on fluorescence assay for bacterial β-lactamases with amyloid fibrils as fluorescence enhancer and visual tool

Herein is described the development of a novel switch-on fluorescence assay for detecting β-lactamases. The fluorescence assay comprises two components: solid beads coated with a β-lactam antibiotic, which is linked to an environment-sensitive fluorophore (dansylaminothiophenol, DTA), and amyloid fibrils of hen lysozyme (acting as fluorescence enhancer and visual tool). In the presence of the clinically significant TEM-1 β-lactamase, the DTA-antibiotic complex on the solid beads is hydrolyzed, thus releasing the DTA dye into solution. The DTA dye is only weakly fluorescent in solution but gives strong green fluorescence upon binding to lysozyme fibrils. These strongly fluorescent DTA-bound fibrils can be easily visualized by the naked eye upon illumination of the sample with a simple UV lamp. The fluorescence assay can detect TEM-1 at low concentration (0.01 nM). In contrast, no observable fluorescence appears when the fluorescence assay is performed on samples without the TEM-1 β-lactamase.     

Synthesis, structural characterization and antibacterial activity of 2,6-diacetylpyridine bis(benzenesulfonohydrazide) Schiff bases and their copper(II) complexes

2,6-Diacetylpyridine bis(benzenesulfonohydrazide) Schiff bases (L¹, L² and L³) and their Cu(II) complexes of the general formula [CuL·H₂O] were synthesized and characterized by various spectroscopic techniques. The crystal structure of [CuL³·(py)]·py was investigated by single crystal X-ray structure analysis. The Cu(II) cation has near square pyramidal, penta-coordinate geometry. The binegatively charged tetradentate Schiff base is asymmetrically coordinated to the Cu(II) ion via the pyridine N atom, the azomethine N atom, the sulfonyl O atom and the deprotonated hydrazine N atom. There is a pyridine molecule apically coordinated to the Cu(II) ion. All the Schiff bases and their copper(II) complexes were screened by the disc diffusion method against multi-drug resistant (MDR) gram-negative and gram-positive bacteria. The minimum inhibitory concentration (MIC) values were also determined. These results show that the antibacterial activity of the Schiff bases against Methicillin-resistant Staphylococcus aureus (MRSA) is enhanced when they are chelated with the copper(II) ion.     

Ferrous Iron Binding Key to Mms6 Magnetite Biomineralisation: A Mechanistic Study to Understand Magnetite Formation Using pH Titration and NMR Spectroscopy

Formation of magnetite nanocrystals by magnetotactic bacteria is controlled by specific proteins which regulate the particles’ nucleation and growth. One such protein is Mms6. This small, amphiphilic protein can self‐assemble and bind ferric ions to aid in magnetite formation. To understand the role of Mms6 during in vitro iron oxide precipitation we have performed in situ pH titrations. We find Mms6 has little effect during ferric salt precipitation, but exerts greatest influence during the incorporation of ferrous ions and conversion of this salt to mixed‐valence iron minerals, suggesting Mms6 has a hitherto unrecorded ferrous iron interacting property which promotes the formation of magnetite in ferrous‐rich solutions. We show ferrous binding to the DEEVE motif within the C‐terminal region of Mms6 by NMR spectroscopy, and model these binding events using molecular simulations. We conclude that Mms6 functions as a magnetite nucleating protein under conditions where ferrous ions predominate.     

多重聚合酶链反应-毛细管电泳-激光诱导荧光法检测三种食源性致病菌

建立了食品中常见致病菌大肠杆菌O157:H7的uidA基因、沙门菌的invA基因和志贺菌的ipaH基因的多重聚合酶链反应（PCR）产物的毛细管电泳快速检测方法。根据这3种致病菌的特异性基因序列设计多重PCR引物,优化PCR扩增反应体系,采用7.0 g/L 甲基纤维素为筛分介质,毛细管电泳-激光诱导荧光检测法同时检测了3种常见致病菌的PCR扩增产物。在优化的多重PCR反应和毛细管筛分电泳条件下,该方法可以同时检测沙门菌、志贺菌和大肠杆菌O157:H7基因的多重PCR扩增产物,22 min内即可完成3种常见致病菌的毛细管电泳检测。迁移时间的相对标准偏差为1.47%~2.07%。与凝胶电泳法比较,该法简便快速,灵敏度高,可用于多种致病菌脱氧核糖核酸的检测,为食品安全提供了一种可靠的快速检测方法。     

Solubilities,thermostabilities and flame retardance behaviour of phosphorus-containing flame retardants and copolymers

Thirteen phosphorus-containing flame retardants were synthesized in this work. The solubilities of flame retardant [(6-oxide-6H-dibenz[c,e][1,2]oxaphosphorin-6-yl)-methyl]-butanedioic acid (DDP) in selected solvents are measured. TGA measurements of the 13 phosphorus-containing flame retardants were carried out and thermal stabilities of three flame-resistant PET (FRPET) resins were investigated. A FRPET incorporated by DDP with terephthalic acid and ethylene glycol reported in literature was also discussed and compared. The thermal stability of the FRPET is improved by the incorporation of phosphorus-containing flame retardants. The LOI values of all phosphorus-containing polyesters are higher than 27%. The improvement of the flame-resistant ability is due to the formation of the char that is not only caused by the existence of phosphorus elements in the resin but also by the relative large number of carbon atoms of the phenyl group in the flame retardants.     

天然橡胶/石墨烯纳米复合材料的制备及耐核辐射性能

采用乳液共混和原位还原法制备了天然橡胶(NR)/还原氧化石墨烯(RGO)纳米复合材料,研究了γ射线辐照对复合材料力学性能和热稳定性的影响.研究结果表明,RGO以少数几层堆叠片层结构均匀分散于NR基体中.RGO的加入可显著提高NR的力学性能和热稳定性,加入质量分数为0.6%的RGO可使材料拉伸强度由(22±1.4)MPa提升至(25±1.1)MPa,质量损失50%对应的温度(T50)升高6.4℃.经200 k Gy的γ射线辐射后,纯NR的拉伸强度和T50分别下降了75%和4.5℃,而NR/RGO-0.6%复合体系仅分别下降了56%和1.2℃.揭示了RGO提高材料耐辐射性能的机理,由于RGO可捕捉猝灭因辐射产生的自由基,从而减弱了辐射老化降解和交联反应的发生.     

原位微区同步辐射X射线荧光和近边吸收谱研究铅耐受细菌吸附-转化铅机理

为了从微观水平研究细菌生物吸附及转化铅机理,利用原位微区同步辐射X射线荧光(μ-SRXRF)及X射线吸收近边结构谱( XANES)研究云南兰坪铅锌矿区农田土壤样品中筛选的铅耐受性细菌吸附铅的分布特征及铅形态转化规律。土壤中具有铅耐受性的菌株主要为Arthrobacter sp.属(节杆菌属),采用μ-SRXRF对其吸附铅的含量进行快速简单直接分析,部分细菌吸附铅的含量高达5925μg/g,富集系数达14.8。XANES结果表明,细菌吸附 Pb 后存在形态为 PbS、(C17 H35 COO)2Pb 和 Pb5(PO4)3Cl 分别占58.0％,22.2％和19.8％,与培养基本身以有机态为主的Pb形态有明显差异,表明培养基中铅被细菌吸附后有向硫化物转化的趋势,这为研究重金属生物有效性的影响因素提供了实验参考。     

Antibacterial activity of cyclo(L-Pro-L-Tyr) and cyclo(D-Pro-L-Tyr) from Streptomyces sp. strain 22-4 against phytopathogenic bacteria

Two bioactive cyclic dipeptides, cyclo(L-Pro-L-Tyr) and cyclo(D-Pro-L-Tyr), were isolated from the culture broth of Streptomyces sp. strain 22-4 and tested against three economically important plant pathogens, Xanthomonas axonopodis pv. citri, Ralstonia solanacearum and Clavibacter michiganensis. Both cyclic dipeptides were active against X. axonopodis pv. citri and R. Solanacearum with MIC of 31.25 μg/mL. No activity could be observed against C. michiganensis.