食品接触表面食源性致病菌生物膜形成及控制研究进展

目的 综述食源性致病菌在食品接触表面形成生物膜的安全性及其控制措施,为解决食品接触表面食源性致病菌形成生物膜的危害问题提供参考.方法 归纳阐述近年来国内外食品接触表面形成食源性致病菌生物膜的危害问题,重点分析生物膜的形成机制、预防和控制措施,以及未来的发展方向.结果 食源性致病菌在食品接触表面形成粘附力大、抗逆性高、适应性广、耐药性强的生物膜结构,使用常规的物理性、化学性生物膜清除方法无法完全去除;广谱微生物源性抗生物膜剂、抗菌包装材料和天然食品级化学杀菌剂具有优良的裂解性、特异性、无毒性、环保性和抗菌性等特点,已成为生物膜清除和控制的有效资源.结论 天然抗菌活性物质、抑菌包装材料和具有生物活性的抗生物膜剂已逐步取代传统的物理性、化学性生物膜清除方法,成为有效保障食品工业生产、消费者食品食用安全的新途径之一.     

TiO<Subscript>2</Subscript>-containing and ZnO-containing borosilicate glass—a novel thin glass with exceptional antibiofilm performances to prevent microfouling

Biofilm formation, also known as microfouling, on indwelling medical devices such as catheters or prosthetic joints causes difficult to treat and recurrent infections. It is also the initial step for biocorrosion of surfaces in aquatic environment. An efficient prevention of microfouling is preferable but the development of antibiofilm surfaces is enormously challenging. Therefore, soda-lime, aluminosilicate, and three borosilicate glasses with different TiO₂ and ZnO compositions were investigated on their feasibility to prevent biofilm formation by standardized in vitro biofilm assays using different pathogenic bacteria. Furthermore, the biocompatibility of these glasses was evaluated using eukaryotic cell lines end erythrocytes. Only two borosilicate glasses, containing TiO₂ and ZnO, showed an increased antibiofilm performance inhibiting biofilm adhesion and formation. The biofilm thickness and area were significantly reduced by over 90?% and characterized by diffuse structures. All tested glass types showed neither cytotoxicity nor hemotoxicity. Therefore, the antibiofilm borosilicate-thin glasses are qualified for surface coatings where biofilms are not desirable such as on medical devices.     

Mechanical robustness of Pseudomonas aeruginosa biofilms

Biofilms grow on various surfaces and in many different environments, a phenomenon that constitutes major problems in industry and medicine. Despite their importance little is known about the viscoelastic properties of biofilms and how these depend on the chemical microenvironment. Here, we find that the mechanical properties of Pseudomonas aeruginosa (P.a.) biofilms are highly robust towards chemical perturbations. Specifically, we observe that P.a. biofilms are able to fully regain their initial stiffness after yielding is enforced, even in the presence of chemicals. Moreover, only trivalent ions and citric acid significantly affect the biofilm elasticity, the first of which also alter the texture of the material. Finally, our results indicate that biofilm mechanics and bacteria viability inside the biofilm are not necessarily linked which suggests that targeting bacteria alone might not be sufficient for biofilm removal strategies.     

Recent advances and future challenges in the use of nanoparticles for the dispersal of infectious biofilms

Increasing occurrence of intrinsically antimicrobial-resistant,human pathogens and the protective biofilm-mode in which they grow,dictates a need for the alternative control of infectious biofilms.Biofilm bacteria utilize dispersal mechanisms to detach parts of a biofilm as part of the biofilm life-cycle during times of nutrient scarcity or overpopulation.We here identify recent advances and future challenges in the development of dispersants as a new infection-control strategy.Deoxyribonuclease(DNase)and other extracellular enzymes can disrupt the extracellular matrix of a biofilm to cause dispersal.Also,a variety of small molecules,reactive oxygen species,nitric oxide releasing compounds,peptides and molecules regulating signaling pathways in biofilms have been described as dispersants.On their own,dispersants do not inhibit bacterial growth or kill bacterial pathogens.Both natural,as well as artificial dispersants,are unstable and hydrophobic which necessitate their encapsulation in smart nanocarriers,like pH-responsive micelles,liposomes or hydrogels.Depending on their composition,nanoparticles can also possess intrinsic dispersant properties.Bacteria dispersed from an infectious biofilm end up in the blood circulation where they are cleared by host immune cells.However,this sudden increase in bacte-rial concentration can also cause sepsis.Simultaneous antibiotic loading of nanoparticles with dispersant properties or combined administration of dispersants and antibiotics can counter this threat.Importantly,biofilm remaining after dispersant administration appears more susceptible to existing antibiotics.Being part of the natural biofilm life-cycle,no signs of"dispersant-resistance"have been observed.Dispersants are therewith promising for the control of infectious biofilms.     

Broad‐spectrum inhibitory effect of green synthesised silver nanoparticles from Withania somnifera (L.) on microbial growth,biofilm and respiration: a putative mechanistic approach

Multi‐drug resistance in pathogenic bacteria has created immense clinical problem globally. To address these, there is need to develop new therapeutic strategies to combat bacterial infections. Silver nanoparticles (AgNPs) might prove to be next generation nano‐antibiotics. However, improved efficacy and broad‐spectrum activity is still needed to be evaluated and understood. The authors have synthesised AgNPs from Withania somnifera (WS) by green process and characterised. The effect of WS‐AgNPs on growth kinetics, biofilm inhibition as well as eradication of preformed biofilms on both gram‐positive and gram‐negative pathogenic bacteria was evaluated. The authors have demonstrated the inhibitory effect on bacterial respiration and disruption of membrane permeability and integrity. It was found that WS‐AgNPs inhibited growth of pathogenic bacteria even at 16 µg/ml. At sub‐minimum inhibitory concentration concentration, there was approximately 50% inhibition in biofilm formation which was further validated by light and electron microscopy. WS‐AgNPs also eradicated the performed biofilms by varying levels at elevated concentration. The bacterial respiration was also significantly inhibited. Interaction of WS‐AgNPs with test pathogen caused the disruption of cell membrane leading to leakage of cellular content. The production of intracellular reactive oxygen species reveals that WS‐AgNPs exerted oxidative stress inside bacterial cell causing microbial growth inhibition and disrupting cellular functions.Inspec keywords: silver, nanoparticles, nanofabrication, nanomedicine, antibacterial activity, biomedical materials, cellular biophysics, microorganisms, biomembranes, electron microscopy, oxidation, biochemistry, permeabilityOther keywords: broad‐spectrum inhibitory effect, green synthesised silver nanoparticles, Withania somnifera (L.), microbial growth, putative mechanistic approach, multidrug resistance, therapeutic strategies, bacterial infections, next generation nanoantibiotics, broad‐spectrum activity, WS‐AgNPs, growth kinetics, biofilm inhibition, gram‐positive pathogenic bacteria, gram‐negative pathogenic bacteria, bacterial respiration, membrane permeability, membrane integrity, subminimum inhibitory concentration concentration, biofilm formation, light pathogenic bacteria, electron microscopy, cell membrane, cellular content leakage, intracellular reactive oxygen species, oxidative stress, microbial growth inhibition, Ag     

具有可控释放能力和可抑制生物膜形成的聚多巴胺/银杂化涂层的钠钙玻璃球

过滤介质作为过滤装置中的关键材料可改变流经其中的水的质量.正确选择过滤介质对于过滤装置的过滤性能至关重要.再生玻璃作为介质过滤装置中硅砂的替代品,具有比硅砂廉价、环境友好、可再生等优点,并且可以根据特定的设计要求将其粉碎成不同的尺寸.然而,水中存在大量的诸如细菌和藻类的微生物,故再生的可循环玻璃介质的过滤效率受到在其表面富集的生物膜的极大限制.本研究中,我们通过在钠钙玻璃球(GS)表面上进行氢氟酸(HF)蚀刻和原位结晶制备了用聚多巴胺(PDA)和银(Ag)纳米颗粒改性的氢氟酸蚀刻玻璃球(PDA-Ag-HF/GSs).银晶体的原位生长、HF蚀刻和PDA涂层的改性赋予了PDA-Ag-HF/GS良好的亲水性.银涂层的改性还使得PDA-Ag-HF/GS具有出色的抗菌性能和较小的球藻附着力,且通过释放Ag离子可抑制微生物的生长.PDA涂层上的邻苯二酚官能团可通过螯合作用调节Ag离子的释放.良好的抗菌性能、抗藻类附着力和Ag离子的受控释放表明,PDA-AgHF/GS涂层可有效抑制材料表面生物膜的形成,为抗生物膜的形成提供了新的策略.     

Structural transformation by electrodeposition on patterned substrates (STEPS): a new versatile nanofabrication method

Arrays of high-aspect-ratio (HAR) nano- and microstructures are of great interest for designing surfaces for applications in optics, bio-nano interfaces, microelectromechanical systems, and microfluidics, but the difficulty of systematically and conveniently varying the geometries of these structures significantly limits their design and optimization for a specific function. This paper demonstrates a low-cost, high-throughput benchtop method that enables a HAR array to be reshaped with nanoscale precision by electrodeposition of conductive polymers. The method-named STEPS (structural transformation by electrodeposition on patterned substrates)-makes it possible to create patterns with proportionally increasing size of original features, to convert isolated HAR features into a closed-cell substrate with a continuous HAR wall, and to transform a simple parent two-dimensional HAR array into new three-dimensional patterned structures with tapered, tilted, anisotropic, or overhanging geometries by controlling the deposition conditions. We demonstrate the fabrication of substrates with continuous or discrete gradients of nanostructure features, as well as libraries of various patterns, starting from a single master structure. By providing exemplary applications in plasmonics, bacterial patterning, and formation of mechanically reinforced structures, we show that STEPS enables a wide range of studies of the effect of substrate topography on surface properties leading to optimization of the structures for a specific application. This research identifies solution-based deposition of conductive polymers as a new tool in nanofabrication and allows access to 3D architectures that were previously difficult to fabricate.     

Physicochemical regulation of biofilm formation

This article reviews the physical and chemical constraints of environments on biofilm formation. We provide a perspective on how materials science and engineering can address fundamental questions and unmet technological challenges in this area of microbiology, such as biofilm prevention. Specifically, we discuss three factors that impact the development and organization of bacterial communities. (1) Physical properties of surfaces regulate cell attachment and physiology and affect early stages of biofilm formation. (2) Chemical properties influence the adhesion of cells to surfaces and their development into biofilms and communities. (3) Chemical communication between cells attenuates growth and influences the organization of communities. Mechanisms of spatial and temporal confinement control the dimensions of communities and the diffusion path length for chemical communication between biofilms, which, in turn, influences biofilm phenotypes. Armed with a detailed understanding of biofilm formation, researchers are applying the tools and techniques of materials science and engineering to revolutionize the study and control of bacterial communities growing at interfaces.     

The contribution of cell-cell signaling and motility to bacterial biofilm formation

Many bacteria grow attached to a surface as biofilms. Several factors dictate biofilm formation, including responses by the colonizing bacteria to their environment. Here we review how bacteria use cell-cell signaling (also called quorum sensing) and motility during biofilm formation. Specifically, we describe quorum sensing and surface motility exhibited by the bacterium Pseudomonas aeruginosa, a ubiquitous environmental organism that acts as an opportunistic human pathogen in immunocompromised individuals. P. aeruginosa uses acyl-homoserine lactone signals during quorum sensing to synchronize gene expression important to the production of polysaccharides, rhamnolipid, and other virulence factors. Surface motility affects the assembly and architecture of biofilms, and some aspects of motility are also influenced by quorum sensing. While some genes and their function are specific to P. aeruginosa, many aspects of biofilm development can be used as a model system to understand how bacteria differentially colonize surfaces.     

Influence of nanocomposite surface coating on biofilm formation in situ

Caries and periodontitis, the most wide-spread oral diseases around the world, are caused by bacterial adherence and biofilm formation onto the natural as well as restored tooth surface. One possible way to prevent the pathogenic consequences of intraoral biofilm formation might be the modification of the tooth surface by application of an anti-adhesive coating that interferes with the bacterial attachment and subsequent bacterial accumulation. The objective of this study was to investigate the effect of an experimental, low surface free energy nano-composite coating material on biofilm formation in situ. For this purpose, an organic/inorganic nano-composite coating (NANOMER, INM, Saarbrücken, Germany) with a surface free energy of 18-20 mJ/m2 was applied to enamel as well as titanium specimens. The nano-composite coated specimens and un-coated controls were attached to removable intraoral splints and carried by volunteers over 24 h in the oral cavity. After intraoral exposure, specimens were processed for transmission electron microscopic analysis. On non-coated enamel and titanium control samples a multi-layer of adherent bacteria was found. In contrast, on nano-composite coated specimens strongly reduced biofilm formation was observed. In most areas of the surface-coated specimens only a 10-20 nm thick electron dense layer of adsorbed salivary proteins with adherent protein agglomerates of 20-80 nm diameter could be detected. In addition, detachment of the adsorbed biofilm from the nano-composite coated surfaces was evident in electron microscopic micrographs. The present investigation provides ultrastructural evidence that it is possible to cover enamel as well as titanium with a nano-composite coating revealing easy-to-clean surface properties that cause reduced biofilm formation and accelerated removal of adherent biofilms under oral conditions.     

Depriving Bacterial Adhesion‐Related Molecule to Inhibit Biofilm Formation Using CeO2‐Decorated Metal‐Organic Frameworks

The formation of bacterial biofilm is one of the causes of antimicrobial resistance, often leading to persistent infections and a high fatality rate. Therefore, there is an urgent need to develop novel and effective strategies to inhibit biofilm formation. Adenosine triphosphate (ATP) plays an important role in bacterial adhesion and biofilm formation through stimulating cell lysis and extracellular DNA (eDNA) release. Herein, a simple and robust strategy for inhibiting biofilm formation is developed using CeO₂‐decorated porphyrin‐based metal‐organic frameworks (MOFs). The function of extracellular ATP (eATP) can be inhibited by CeO₂ nanoparticles, leading to the disruption of the initial adhesion of bacteria. Furthermore, planktonic bacteria can be killed by cytotoxic reactive oxygen species (ROS) generated by MOFs. As a consequence, the synergic effect of eATP deprivation and ROS generation presents excellent capacity to prevent biofilm formation, which may provide a new direction for designing flexible and effective biofilm‐inhibiting systems.     

Antibacterial Performance of a Cu-bearing Stainless Steel against Microorganisms in Tap Water

Tap water is one of the most commonly used water resources in our daily life. However, the increasing water contamination and the health risk caused by pathogenic bacteria, such as Staphylococcus aureus and Escherichia coli have attracted more attention. The mutualism of different pathogenic bacteria may diminish antibacterial effect of antibacterial agents. It was found that materials used for making pipe and tap played one of the most important roles in promoting bacterial growth. This paper is to report the performance of an innovative type 304 Cu-bearing stainless steel(304Cu SS) against microbes in tap water. The investigation methodologies involved were means of heterotrophic plate count, contact angle measurements, scanning electron microscopy for observing the cell and subtract surface morphology,atomic absorption spectrometry for copper ions release study, and confocal laser scanning microscopy used for examining live/dead bacteria on normal 304 stainless steel and 304 Cu SS. It was found that the surface free energy varied after being immersed in tap water with polar component and Cu ions release.The results showed 304 Cu SS could effectively kill most of the planktonic bacteria(max 95.9% antibacterial rate), and consequently inhibit bacterial biofilms formation on the surface, contributing to the reduction of pathogenic risk to the surrounding environments.     

Electrical spiking in bacterial biofilms

In nature, biofilms are the most common form of bacterial growth. In biofilms, bacteria display coordinated behaviour to perform specific functions. Here, we investigated electrical signalling as a possible driver in biofilm sociobiology. Using a multi-electrode array system that enables high spatio-temporal resolution, we studied the electrical activity in two biofilm-forming strains and one non-biofilm-forming strain. The action potential rates monitored during biofilm-forming bacterial growth exhibited a one-peak maximum with a long tail, corresponding to the highest biofilm development. This peak was not observed for the non-biofilm-forming strain, demonstrating that the intensity of the electrical activity was not linearly related to the bacterial density, but was instead correlated with biofilm formation. Results obtained indicate that the analysis of the spatio-temporal electrical activity of bacteria during biofilm formation can open a new frontier in the study of the emergence of collective microbial behaviour.     

3-D Numerical Simulations of Biofilm Flows

We study the biofilm-flow interaction resulting in biofilm growth and deformation in a water channel in a 3-D setting using the phase field model developed recently [28, 29]. In this biofilm model, the biofilm made up of the EPS, bacteria and solvent is tracked using a biofilm volume fraction which vanishes outside the biofilm region. The interface between the biofilm and the solvent is marked by the zero level surface of the volume fraction measured from the biofilm to the solvent. The growth of the biofilm and the solvent-biofilm interaction with the top nutrient feeding condition is simulated in the viscous regime (growth regime) of the biofilm-solvent mixture flow. In quiescent flows, the model predicts growth patterns consistent with experimental findings for single or multiple adjacent biofilm colonies, in which the known mushroom shape growth pattern is obtained. Shear induced deformation in biofilms is simulated in a shear cell, providing a viable numerical evidence for using simulation tool to study biofilm growth and interaction dynamics in aqueous environment.     

In vitro and in vivo microbial adhesion and growth on argon plasma-treated silicone rubber voice prostheses

Patients who undergo a total laryngectomy usually receive a silicone rubber voice prosthesis for voice rehabilitation. Unfortunately, biofilm formation on the esophageal side of voice prostheses limits their lifetime to 3–4 mon on average. The effects of repeated argon plasma treatment of medical grade, hydrophobic silicone rubber on in vitro adhesion and growth of bacteria and yeasts isolated from voice prostheses, as well as in vivo biofilm formation are presented here. In vitro experiments demonstrated that initial microbial adhesion over a 4 h time span to plasma-treated, hydrophilized, silicone rubber was generally less than on original, hydrophobic silicone rubber, both in the absence and presence of a salivary conditioning film on the biomaterial. Growth studies over a time period of 14 d at 37°C in a modified Robbins device, showed that fewer Candida cells adhered on plasma-treated, hydrophilized silicone rubber as compared to on original, hydrophobic silicone rubber. For the in vivo evaluation of biofilm formation on plasma-treated silicone rubber voice prostheses, seven laryngectomized patients received a partly hydrophilized Groningen Button voice prosthesis for a planned evaluation period of 4 wk. After removal of the voice prostheses, the border between the hydrophilized and the original, hydrophobic side of the prostheses was clearly visible. However, biofilm formation was, unexpectedly, less on the original, hydrophobic sides, although the microbial compositions of the biofilms on both sides were not significantly different. Summarizing, this study demonstrates that in vitro microbial adhesion and growth on silicone rubber can be reduced by plasma treatment, but in vivo biofilm formation on silicone rubber voice prostheses is oppositely enhanced by hydrophilizing the silicone rubber surface. Nevertheless, from the results of this study the important conclusion can be drawn that in vivo biofilm formation on voice prostheses is controlled by the hydrophobicity of the biomaterials surface used. © 1998 Chapman & Hall     

抗菌剂对聚乙烯表面生物被膜的抑制

采用共混方法在低密度聚乙烯中分别加入银系抗菌剂和Triclosan制备出两种改性聚乙烯,研究了材料的抗菌性能和抗菌剂对试样表面生物被膜的抑制作用.结果表明,两种改性聚乙烯对大肠杆菌和金黄色葡萄球菌都有良好的抗菌效果;细菌生物被膜的形成主要包括粘附、繁殖和成熟3个阶段,添加Triclosan的试样可以在生物被膜成熟前显著杀灭样品表面的细菌,阻止细菌在其表面粘附和繁殖,从而抑制生物被膜形成.空白试样和银系抗菌剂改性试样表面均有大量细菌粘附并形成生物被膜.     

Zirconium oxide nanotube surface prompts increased osteoblast functionality and mineralization

Electrochemical formation of tunable nanoscale oxide layers on biomedical metallic surfaces has recently drawn much attention in biomaterials research. In this study, we report on the cellular response to a unique vertically aligned, laterally spaced nanotube nanostructure made of zirconium oxide (ZrO₂) fabricated by anodization. The growth, morphology, and functionality of osteoblasts cultured on ZrO₂ nanotubes have been investigated. The initial adhesion and spreading was considerably improved on the nanotube surface as compared to a flat zirconium (Zr) surface without a nanostructure. The morphology of the adhered cells on the nanotube surface elicited a highly organized cytoskeleton with crisscross patterned actin, which was lacking on the flat Zr. Increased alkaline phosphatase activity levels and the formation of calcified extracellular matrix implied improved osteoblast functionality and mineralization on the nanotube substrate. This in vitro study suggests that the ZrO₂ nanotubes provided an enhanced osteoblast response and demonstrated their apparent role in providing a platform for bone growth.     

Local pH variation as an initial step in bacterial surface-sensing and biofilm formation

The development of surface-associated layers of matrix-embedded microbial populations, called biofilms, is a serious problem in many medical and industrial settings. These contaminations are difficult to eradicate because of the high resistance level acquired by the cells in their particular environment. From the very beginning of the colonization process, modifications of gene expression are observed and could, at least partially, explain biofilm resistance. In order to develop anti-biofilm molecules and surface treatments, it is of pivotal importance to identify the physico-chemical parameters which activate the sensor systems of pioneering microbes when they come into contact with a surface. The aim of our study was to examine the pH variations in the local micro-environment created between the cell layer and the surface after bacteria adhesion. Using an ion-sensitive field effect transistor (ISFET), as a substratum, colonized by a curli hyperproducing strain known to form biofilms, we observed that the evolution of the pH change was significantly different in the micro-compartment in contact with the electrochemical sensor compared to that within the liquid phase.     

Dynamic Sessile‐Droplet Habitats for Controllable Cultivation of Bacterial Biofilm

Bacterial biofilms play essential roles in biogeochemical cycling, degradation of environmental pollutants, infection diseases, and maintenance of host health. The lack of quantitative methods for growing and characterizing biofilms remains a major challenge in understanding biofilm development. In this study, a dynamic sessile‐droplet habitat is introduced, a simple method which cultivates biofilms on micropatterns with diameters of tens to hundreds of micrometers in a microfluidic channel. Nanoliter plugs are utilized, spaced by immiscible carrier oil to initiate and support the growth of an array of biofilms, anchored on and spatially confined to the micropatterns arranged on the bottom surface of the microchannel, while planktonic or dispersal cells are flushed away by shear force of aqueous plugs. The performance of the aforementioned method of cultivating biofilms is demonstrated by Pseudomonas aeruginosa PAO1 and its derived mutants, and quantitative antimicrobial susceptibility testing of PAO1 biofilms. This method could significantly eliminate corner effects, avoid microchannel clogging, and constrain the growth of biofilms for long‐term observations. The controllable sessile droplet‐based biofilm cultivation presented in this study should shed light on more quantitative and long‐term studies of biofilms, and open new avenues for investigation of biofilm attachment, growth, expansion, and eradication.     

One‐Step Synthesis of Epoxy Group‐Terminated Organosilica Nanodots: A Versatile Nanoplatform for Imaging and Eliminating Multidrug‐Resistant Bacteria and Their Biofilms

Multidrug‐resistant bacteria (MRB) and their biofilms, both of which develop high levels of drug tolerance, cause severe threats to global health. This study demonstrates that biocompatible fluorescent silicon‐containing nanodots can be a multifunctional platform for simultaneously imaging and eliminating MRB and their biofilms. Ultrasmall epoxy group (oxirane)‐functionalized organosilica nanodots (OSiNDs) with a high photoluminescence quantum yield of ≈31% are synthesized via a simple one‐step hydrothermal treatment of an epoxy group‐containing silane molecule, 3‐glycidoxypropyltrimethoxysilane, and an organic dye, rose bengal. The resultant OSiNDs can be employed as a universal imaging reagent for visualizing various bacteria/biofilms, including MRB and their biofilms. Moreover, the epoxy group‐terminated OSiNDs can be conjugated with amine‐containing reagents only via the simple stirring of the mixtures at an elevated temperature (e.g., 60 °C) for several hours (e.g., 3 h) without the addition of activating reagents. The amine‐containing antibiotic vancomycin (Van) can thus be easily conjugated with the OSiNDs, and the obtained OSiNDs‐Van can successfully inhibit the growth of MRB and even eliminate their biofilms. Collectively, the present work may give new impetus to the development of novel antibacterial and anti‐biofilm agents for overcoming the drug resistance of bacteria.