碳钢在微生物介质中的腐蚀电化学行为

硫酸盐还原菌（SRB）生长代谢，溶液pH值从7．5下降到5．8，并在碳钢表面形成生物膜，改变了生物膜下碳钢表面的微环境，促进碳钢点蚀的形成，清除表面的腐蚀产物后，碳钢基体表面为分布不均匀的溃斑，发生严重的点蚀行为，而RB的活性和生物膜的结构诱发碳钢腐蚀的形成和生长，在微生物快速生长过程中出现明显的氢扩散电流峰，加剧碳钢的渗氢行为，采用电化学阻抗（EIS）研究了碳钢在微生物腐蚀过程中生物膜的结构与腐蚀相应的变化关系。     

再生水环境中304不锈钢生物膜腐蚀电化学特征

研究了以再生水作为循环冷却系统补水的北京某热电厂冷却塔底粘泥中分离纯化培养出来的硫酸盐还原菌(SRB)生长特性.采用原子力显微镜(AFM)、扫描电镜(SEM)、能谱分析仪(EDS)和电化学交流阻抗(EIS)方法研究了304不锈钢(SS304)表面生物膜特征及其主要成分和不锈钢/生物膜界面电化学行为.结果显示,再生水环境下304不锈钢表面形成的生物膜是由吸附的SRB菌体及以含碳有机物为主的胞外聚合物和FeS腐蚀产物构成.浸泡前期(前7 d)SS304电极表面阻抗值主要由SS304表面钝化膜的贡献;浸泡后期(14 d后),电极体系阻抗值由不锈钢表面钝化膜和生物膜共同贡献.     

Quantitative and morphological analysis of biofilm formation on self-assembled monolayers

In spite of intensive studies over the past two decades, the influence of surface properties on bacterial adhesion and biofilm formation remains unclear, particularly on late steps. In order to contribute to the elucidation of this point, we compared the impact of two different substrates on the formation of bacterial biofilm, by analysing bacterial amount and biofilm structure on hydrophilic and hydrophobic surfaces. The surfaces were constituted by NH₂- and CH₃-terminated self-assembled monolayers (SAMs) on silicon wafers, allowing to consider only the surface chemistry influence because wafers low roughness. A strain of Escherichia coli K12, able to produce biofilm on abiotic surfaces, was grown with culture durations varying from 4 h to 336 h on both types of substrates. The amount of adhered bacteria was determined after detachment by both photometry at 630 nm and direct counting under light microscope, while the spatial distribution of adhered bacteria was observed by fluorescence microscopy. A general view of our results suggests a little influence of the surface chemistry on adherent bacteria amount, but a clear impact on dynamics of biofilm growth as well as on biofilm structure. This work points out how surface chemistry of substrates can influence the bacterial adhesion and the biofilm formation.     

Accelerated anaerobic corrosion of electroactive sulfate-reducing bacteria by electrochemical impedance spectroscopy and chronoamperometry

Many microbial species can use cathodes as electron donors for metabolism. This direct electron transfer (DET) pathway has rarely been proposed in biocorrosion processes. DET from Q235 carbon steel to the sulfate-reducing bacteria (SRB) Desulfovibrio caledoniensis and its effect on the corrosion behavior of carbon steel were investigated in the present study. Electroactive SRB biofilm was found to play a key role in the ennoblement of the corrosion potential (E_corr) and in the acceleration of the corrosion rate, indicating that SRB mainly affected the cathodic reaction of low carbon steel corrosion. In addition, SRB biofilm obtained electrons from carbon steel electrode polarized at − 0.74 V. These findings present new evidence for DET between SRB biofilm and carbon steels.     

Determination of the van der Waals, electron donor and electron acceptor surface tension components of static Gram-positive microbial biofilms

A large number of studies have shown the influence of the physico-chemical properties of a surface on microbial adhesion phenomenon. In this study, we considered that the presence of a bacterial biofilm may be regarded as a “conditioning film” that may modify the physico-chemical characteristics of the support, and thus the adhesion capability of planktonic micro-organisms coming into contact with this substratum. In this context, we adapted a protocol for biofilm formation that allows, under our experimental conditions, contact angle measurements, the reference method to determine the energetic surface properties of a substratum. This made it possible to determine the van der Waals, electron acceptor and electron donor properties of static biofilms grown at 25°C on stainless-steel slides with six Gram-positive bacteria isolated in dairy plants. A variance analysis indicated significant effects (P<0.05) of the bacterial strains and of the physiological state of the micro-organisms (planktonic or sessile) on the contact angles. To link the energetic properties of the six biofilms with direct adhesion experiments, we measured the affinity of fluorescent carboxylate-modified polystyrene beads for the different biofilm surfaces. The results correlated best with the electron-acceptor components of the biofilm surface energies, stressing the importance of Lewis acid–base interactions in adhesion mechanisms.     

Intracellularly grown gold nanoislands as SERS substrates for monitoring chromate,sulfate and nitrate localization sites in remediating bacteria biofilms by Raman chemical imaging

Understanding the chemical composition of biofilm matrices is vital in different fields of biology such as surgery, dental medicine, synthetic grafts and bioremediation. The knowledge of biofilm development, composition, active reduction sites and remediation efficacy will help in the development of effective solutions and evaluation of remediating approaches prior to implementation. Surface-enhanced Raman spectroscopy (SERS) based imaging is an invaluable tool to obtain an understanding of the remediating efficacy of microorganisms and its role in the formation of organic and inorganic compounds in biofilms. We demonstrate for the first time, the presence of chromate, sulfate, nitrate and reduced trivalent chromium in soil biofilms. In addition, we demonstrate that SERS imaging was able to validate two observations made by previous studies on chromate/sulfate and chromate/nitrate interactions in Shewanella oneidensis MR-1 biofilms. Additionally, we show a detailed Raman mapping based evidence of the existence of chromate–sulfate competition for cellular entry. Subsequently, we use Raman mapping to study the effect of nitrate on chromate reduction. The findings presented in this paper are among the first to report – detection of multiple metallic ions in bacterial biofilms using intracellular SERS substrates. Such a detailed characterization of biofilms using gold nanoislands based SERS mapping substrate can be extended to study cellular localization of other metallic ions and chemical species of biological and toxicological significance and their effect on reduction reactions in bacterial biofilms.     

生物膜和腐蚀产物膜对A3钢的腐蚀作用研究

应用电化学方法和表面分析技术(AFM和SEM)研究硫酸盐还原菌(SRBB)(生物环境)及其腐蚀产物(非生物环境)对A3钢腐蚀行为的影响以及A3钢在两种不同环境下的腐蚀特征.结果表明:于不同时期生成的微生物膜和腐蚀产物膜,对材料的腐蚀起着不同的作用.生成的生物膜越厚越容易剥落,而不均匀的微生物膜将引起材料的局部腐蚀.在非生物环境中生成的腐蚀产物膜比在生物环境中生成的膜更加紧密地黏附于金属的表面.     

碳钢在生物膜和硫化物膜下的电化学腐蚀行为

应用丝束电极技术比较了SRB生物膜以及硫化物膜对Q2 35碳钢腐蚀过程的影响机制 ,采用电位、电流扫描技术测试了生物膜和FeS膜下的碳钢腐蚀不均匀性特征 ,发现由于膜的导电性致使表面电位扫描已不能作为膜下局部腐蚀的判据 .动电位扫描表明无氧近中性溶液中 ,硫化物膜对碳钢具有一定保护作用 .电化学阻抗谱显示 ,硫化物膜电容增加缓慢 ,其极化电阻Rp 随时间呈先增后降的趋势 .与硫化物膜相比 ,生物膜表现出极大的电容 (10 4 ～ 10 5μF/cm2 ) ,且膜电容随时间呈S型增加 ,而极化电阻Rp 则呈指数下降 ,由此可知生物膜加速了腐蚀     

Development of Chitosan-Based Surfaces to Prevent Single- and Dual-Species Biofilms of Staphylococcus aureus and Pseudomonas aeruginosa

Implantable medical devices (IMDs) are susceptible to microbial adhesion and biofilm formation, which lead to several clinical complications, including the occurrence of implant-associated infections. Polylactic acid (PLA) and its composites are currently used for the construction of IMDs. In addition, chitosan (CS) is a natural polymer that has been widely used in the medical field due to its antimicrobial and antibiofilm properties, which can be dependent on molecular weight (Mw). The present study aims to evaluate the performance of CS-based surfaces of different Mw to inhibit bacterial biofilm formation. For this purpose, CS-based surfaces were produced by dip-coating and the presence of CS and its derivatives onto PLA films, as well surface homogeneity were confirmed by contact angle measurements, Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). The antimicrobial activity of the functionalized surfaces was evaluated against single- and dual-species biofilms of Staphylococcus aureus and Pseudomonas aeruginosa. Chitosan-based surfaces were able to inhibit the development of single- and dual-species biofilms by reducing the number of total, viable, culturable, and viable but nonculturable cells up to 79%, 90%, 81%, and 96%, respectively, being their activity dependent on chitosan Mw. The effect of CS-based surfaces on the inhibition of biofilm formation was corroborated by biofilm structure analysis using confocal laser scanning microscopy (CLSM), which revealed a decrease in the biovolume and thickness of the biofilm formed on CS-based surfaces compared to PLA. Overall, these results support the potential of low Mw CS for coating polymeric devices such as IMDs where the two bacteria tested are common colonizers and reduce their biofilm formation.     

Spring constants and adhesive properties of native bacterial biofilm cells measured by atomic force microscopy

Bacterial biofilms were imaged by atomic force microscopy (AFM), and their elasticity and adhesion to the AFM tip were determined from a series of tip extension and retraction cycles. Though the five bacterial strains studied included both Gram-negative and -positive bacteria and both environmental and laboratory strains, all formed simple biofilms on glass surfaces. Cellular spring constants, determined from the extension portion of the force cycle, varied between 0.16+/-0.01 and 0.41+/-0.01 N/m, where larger spring constants were measured for Gram-positive cells than for Gram-negative cells. The nonlinear regime in the extension curve depended upon the biomolecules on the cell surface: the extension curves for the smooth Gram-negative bacterial strains with the longest lipopolysaccharides on their surface had a larger nonlinear region than the rough bacterial strain with shorter lipopolysaccharides on the surface. Adhesive forces between the retracting silicon nitride tip and the cells varied between cell types in terms of the force components, the distance components, and the number of adhesion events. The Gram-negative cells' adhesion to the tip showed the longest distance components, sometimes more than 1 microm, whereas the shortest distance adhesion events were measured between the two Gram-positive cell types and the tip. Fixation of free-swimming planktonic cells by NHS and EDC perturbed both the elasticity and the adhesive properties of the cells. Here we consider the biochemical meaning of the measured physical properties of simple biofilms and implications to the colonization of surfaces in the first stages of biofilm formation.     

Microbiologically-influenced corrosion of the electroless-deposited NiP-TiNi – Coating

In this study, we reveal the microbiologically influenced corrosion (MIC) behavior of the new electroless NiP-TiNi nanocomposite coating in simulated seawater using the electrochemical impedance spectroscopy (EIS) technique after different periods of incubation time (7, 10, 14, 21, 28 days) in a sulfate-reducing bacteria (SRB) medium. The biofilm formation and the corrosion products were characterized using the scanning electron microscope (SEM) and X-ray photoelectron spectroscopy (XPS). The EIS results revealed the carbon steel (CS)/NiP-TiNi and NiP-TiNi/SRB biofilm interfaces' characteristics after different incubation times in the SRB media. EIS measurements revealed that the NiP-TiNi nanocomposite coating's MIC resistances are superior relative to API X80 carbon steel and a TiNi-free NiP coating, with ∼93% of corrosion inhibition efficiency after 28 days of incubation.     

Correlation between Enterococcus faecalis biofilms development stage and quantitative surface roughness using atomic force microscopy.

Biofilms are assemblages of microorganisms and their associated extracellular products at an interface and typically with an abiotic or biotic surface. The study of the morphology of biofilms is important because they are associated with processes of biofouling, corrosion, catalysis, pollutant transformation, dental caries, drug resistance, and so forth. In the literature, biofilms have been examined by atomic force microscopy (AFM), which has proven to be a potent tool to study different aspects of the biofilm development on solid surfaces. In this work, we used AFM to investigate topographical changes during the development process of Enterococcus faecalis biofilms, which were generated on sterile cellulose nitrate membrane (CNM) filters in brain heart infusion (BHI) broth agar blood plates after 24, 36, 72, 192, and 360 h. AFM height images showed topographical changes due to biofilm development, which were used to characterize several aspects of the bacterial surface, such as the presence of extracellular polymeric substance, and the biofilm development stage. Changes in the development stage of the biofilm were shown to correlate with changes in the surface roughness as quantified through the mean roughness.     

氧化亚铁硫杆菌作用下A3钢的腐蚀行为

采用微生物学方法、电化学方法和表面分析技术研究了A3钢在氧化亚铁硫杆菌中的腐蚀特征和生物膜形貌, 分析了微生物的存在对A3钢电化学行为的影响. 极化曲线测试结果表明, 细菌的存在使浸泡20天后A3钢电极的自腐蚀电位升高, 腐蚀电流密度增大. 原子力显微镜测试结果表明, 浸泡7天的A3钢表面生物膜分布不均匀, 从而引发点蚀萌生. 由于细菌的代谢作用以及A3钢表面腐蚀产物与生物膜的特殊形貌特征, 浸泡在菌液中7天后的A3钢表面有点蚀出现; 随着时间的延长, A3钢表面的点蚀坑深度增大且数量增多, 氧化亚铁硫杆菌的存在使A3钢的局部腐蚀程度加剧.     

Microrheology of bacterial biofilms in vitro: Staphylococcus aureus and Pseudomonas aeruginosa

The rheology of bacterial biofilms at the micron scale is an important step to understanding the communal lifecycles of bacteria that adhere to solid surfaces, as it measures how they mutually adhere and desorb. Improvements in particle-tracking software and imaging hardware have allowed us to successfully employ particle-tracking microrheology to measuring single-species bacterial biofilms, based on Staphlococcus aureus and Pseudomonas aeruginosa. By tracking displacements of the cells at a range of timescales, we separate active and thermal contributions to the cell motion. The S. aureus biofilms in particular show power-law rheology, in common with other dense colloidal suspensions. By calculating the mean compliance of S. aureus biofilms, we observe them becoming less compliant during growth, and more compliant during starvation. The biofilms are rheologically inhomogeneous on the micron scale, as a result of the strength of initial adhesion to the flow cell surface, the arrangement of individual bacteria, and larger-scale structures such as flocs of P. aeruginosa. Our S. aureus biofilms became homogeneous as a function of height as they matured: the rheological environment experienced by a bacterium became independent of how far it lived from the flow cell surface. Particle-tracking microrheology provides a quantitative measure of the "strength" of a biofilm. It may therefore prove useful in identifying drug targets and characterizing the effect of specific molecular changes on the micron-scale rheology of biofilms.     

Force measurements of bacterial adhesion on metals using a cell probe atomic force microscope

The adhesion of microbial cells to metal surfaces in aqueous media is an important phenomenon in both the natural environment and engineering systems. The adhesion of two anaerobic sulfate-reducing bacteria (Desulfovibrio desulfuricans and a local marine isolate) and an aerobe (Pseudomonas sp.) to four polished metal surfaces (i.e., stainless steel 316, mild steel, aluminum, and copper) was examined using a force spectroscopy technique with an atomic force microscope (AFM). Using a modified bacterial tip, the attraction and repulsion forces (in the nano-Newton range) between the bacterial cell and the metal surface in aqueous media were quantified. Results show that the bacterial adhesion force to aluminum is the highest among the metals investigated, whereas the one to copper is the lowest. The bacterial adhesion forces to metals are influenced by both the electrostatic force and metal surface hydrophobicity. It is also found that the physiological properties of the bacterium, namely the bacterial surface charges and hydrophobicity, also have influence on the bacteria-metal interaction. The adhesion to the metals by Pseudomonas sp. and D. desulfuricans was greater than by the marine SRB isolate. The cell-cell interactions show that there are strong electrostatic repulsion forces between bacterial cells. Cell probe atomic force microscopy has provided some useful insight into the interactions of bacterial cells with the metal surfaces.     

Continuous nondestructive monitoring of <Emphasis Type="Italic">Bordetella pertussis</Emphasis> biofilms by Fourier transform infrared spectroscopy and other corroborative techniques

This work describes the application of several analytical techniques to characterize the development of Bordetella pertussis biofilms and to examine, in particular, the contribution of virulence factors in this development. Growth of surface-attached virulent and avirulent B. pertussis strains was monitored in continuous-flow chambers by techniques such as the crystal violet method, and nondestructive methodologies like fluorescence microscopy and Fourier transform (FT) IR spectroscopy. Additionally, B. pertussis virulent and avirulent strains expressing green fluorescent protein were grown adhered to the base of a glass chamber of 1-μm thickness. Three-dimensional images of mature biofilms, acquired by confocal laser scanning microscopy, were quantitatively analysed by means of the computer program COMSTAT. Our results indicate that only the virulent (Bvg⁺) phase of B. pertussis is able to attach to surfaces and develop a mature biofilm. In the virulent phase these bacteria are capable of producing a biofilm consisting of microcolonies of approximately 200 μm in diameter and 24 μm in depth. FTIR spectroscopy allowed us not only to follow the dynamics of biofilm growth through specific biomass and biofilm marker absorption bands, but also to monitor the maturation of the biofilm by means of the increase of the carbohydrate-to-protein ratio.     

A Multinuclear Metal Complex Based DNase‐Mimetic Artificial Enzyme: Matrix Cleavage for Combating Bacterial Biofilms

Extracellular DNA (eDNA) is an essential structural component during biofilm formation, including initial bacterial adhesion, subsequent development, and final maturation. Herein, the construction of a DNase‐mimetic artificial enzyme (DMAE) for anti‐biofilm applications is described. By confining passivated gold nanoparticles with multiple cerium(IV) complexes on the surface of colloidal magnetic Fe₃O₄ /SiO₂ core/shell particles, a robust and recoverable artificial enzyme with DNase‐like activity was obtained, which exhibited high cleavage ability towards both model substrates and eDNA. Compared to the high environmental sensitivity of natural DNase in anti‐biofilm applications, DMAE exhibited a much better operational stability and easier recoverability. When DMAE was coated on substratum surfaces, biofilm formation was inhibited for prolonged periods of time, and the DMAE excelled in the dispersion of established biofilms of various ages. Finally, the presence of DMAE remarkably potentiated the efficiency of traditional antibiotics to kill biofilm‐encased bacteria and eradiate biofilms.     

Effect of antimicrobial peptide (AMP)–tethered stainless steel surfaces on the bacterial membrane

Bacterial colonization leading to biofilm formation on surfaces induces industry-related as well as health care–associated infections worldwide. Emerging antibiotic-resistant microbes and related persistent infections due to adherent biofilm formation on surfaces have urged the need of effective alternative solutions to eradicate prevailing problems. Antimicrobial peptides are considered as potential candidates with distinguished characteristics, namely, broad-spectrum antimicrobial activity, low propensity toward pathogen resistance, and low immune response. In this study, we immobilize an in-house–designed peptide, KLLLRLRKLLRR (KLR), using a 2-step functionalization strategy onto stainless steel (SS) surfaces. SS is amino-silanized using (3-aminopropyl) triethoxysilane followed by tethering of KLR on it via formation of the amide bond. KLR-coated SS surfaces show nearly 95–100% reduction in bacterial colonization in vitro as obtained from antibacterial susceptibility testing while being non-toxic to mammalian cells. The coating strategy does not affect the microstructure of the SS surfaces. These findings demonstrate that this tethering process is able to produce excellent antibacterial surfaces.     

Corrosion Behavior of Steel A3 Influenced by Thiobacillus Ferrooxidans

The electrochemical measurement and surface analysis methods were employed to investigate the corrosion behavior of steel A3 influenced by Thiobacillus ferrooxidans (T.f). Polarization curve results indicated that the presence of Thiobacillus ferrooxidans resulted in higher corrosion potential of the electrode and obviously accelerated the corrosion current density. Atomic force microscope (AFM) results showed that asymmetric biofilms adhered to the surface of steel A3 after 7 days of exposure. The scanning electron microscopy (SEM) results showed that pitting appeared on the surface of steel A3 after 7 days of exposure in T.f solution, which was induced by the metabolism of bacteria and the morphology of the deposit. Pitting holes of steel A3 in T.f solution were deeper after 20 days of exposure. The presence of Thiobacillus ferrooxidans aggravated the localized corrosion of A3 steel.     

A3钢在氧化亚铁硫杆菌、链霉菌及其混合菌体系中的腐蚀行为

采用电化学方法和表面分析技术研究了A3钢在链霉菌(Stremptomyces)和氧化亚铁硫杆菌(Thiobacillus fer-rooxidans,T.f)单独及共同作用下的腐蚀行为.结果表明,试样在不同的含菌腐蚀体系中浸泡7d后,表面生成了不均匀的生物膜层,并表现出各不相同的形貌特征;A3钢在氧化亚铁硫杆菌和链霉菌单种菌作用下发生了局部腐蚀,而混合菌体系中的试样发生均匀腐蚀;混合菌体系中A3钢的腐蚀失重速率介于两种菌单独存在时的腐蚀失重速率之间;试样浸泡14d后的电化学阻抗谱(EIS)测试结果表明,A3钢电极在氧化亚铁硫杆菌-链霉菌混合菌体系中的阻抗值介于两种单菌腐蚀体系之间.以上研究结果表明,A3钢在链霉菌体系中腐蚀最重,混合菌体系其次,氧化亚铁硫杆菌体系中腐蚀最轻.