Biofilm Bridges Forming Structural Networks on Patterned Lubricant-Infused Surfaces.

Despite many decades of research, biofilm architecture and spreading mechanisms are still not clear because of the heterogenous 3D structure within biofilms. Here, patterned "slippery" lubricant-infused porous surfaces are utilized to study biofilm structure of Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Staphylococcus aureus. It is found that bacteria are able to spread over bacteria-repellent lubricant-infused regions by using a mechanism, termed "biofilm bridges". Here, it is demonstrated that bacteria use bridges to form interconnected networks between distant biofilm colonies. Detailed structure of bridges shows a spatial distribution of bacteria with an accumulation of respiratory active bacteria and biomass in the bridges. The core-shell structure of bridges formed by two-species mixed population is illustrated. It is demonstrated that eDNA and nutrients have a strong effect on biofilm bridges formation. Thus, it is believed that biofilm bridging is important to reveal the structure and communication within biofilms.

Study of Biofilm Growth on Slippery Liquid-Infused Porous Surfaces Made from Fluoropor.

Undesired growth of biofilms represents a fundamental problem for all surfaces in long-term contact with aqueous media. Mature biofilms resist most biocide treatments and often are a pathogenic threat. One way to prevent biofilm growth on surfaces is by using slippery liquid-infused porous surfaces (SLIPS). SLIPS consist of a porous substrate which is infused with a lubricant immiscible with the aqueous medium in which the bacteria are suspended. Because of the lubricant, bacteria cannot attach to the substrate surface and thus formation of the biofilm is prevented. For this purpose, we manufactured substrates with different porosity and surface roughness values via UV-initiated free-radical polymerization in Fluoropor. Fluoropor is a class of highly fluorinated bulk-porous polymers with tunable porosity, which we recently introduced. We investigated the growth of the biofilm on the substrates, showing that a reduced surface roughness is beneficial for the reduction of biofilm growth. Samples of low roughness effectively reduced Pseudomonas aeruginosa biofilm growth for 7 days in a flow chamber experiment. The low-roughness samples also become transparent when infused with the lubricant, making such surfaces ideal for real-time observation of biofilm growth by optical examination.

Patterned SLIPS for the Formation of Arrays of Biofilm Microclusters with Defined Geometries.

Biofilms represent an immense problem in medicine due to their strong drug-resistant properties and inherent stress-response activities. Due to the inhomogeneous and very complex architectures of large biofilm aggregates, biofilm studies often suffer from low reproducibility. In this study, an approach to form arrays of homogeneous biofilm microclusters with defined 2D geometries is presented. The method is based on the formation of water-infused hydrophilic porous polymer areas with precise geometries separated by "slippery" lubricant-infused porous surface (SLIPS). Due to the SLIPS' biofilm repellent properties, multiple identical 3D biofilm clusters are formed in the hydrophilic patches that can be used for biofilm screening. Formation of biofilm microcluster arrays of different bacterial strains of Pseudomonas aeruginosa on the SLIPS micropatterns is investigated. Critical parameters influencing minimal adhesive regions for biofilm attachment and minimal SLIPS dimensions to avoid biofilm adhesion are studied. The ability to produce arrays of biofilm microclusters with highly uniform, well-defined shapes opens an opportunity to study interactions of biofilms with various medically relevant factors with a better reproducibility and to investigate the complex biofilm architecture, heterogeneity, and interactions between biofilm subpopulations.

Multi-channel microfluidic biosensor platform applied for online monitoring and screening of biofilm formation and activity.

Bacterial colonization of surfaces and interfaces has a major impact on various areas including biotechnology, medicine, food industries, and water technologies. In most of these areas biofilm development has a strong impact on hygiene situations, product quality, and process efficacies. In consequence, biofilm manipulation and prevention is a fundamental issue to avoid adverse impacts. For such scenario online, non-destructive biofilm monitoring systems become important in many technical and industrial applications. This study reports such a system in form of a microfluidic sensor platform based on the combination of electrical impedance spectroscopy and amperometric current measurement, which allows sensitive online measurement of biofilm formation and activity. A total number of 12 parallel fluidic channels enable real-time online screening of various biofilms formed by different Pseudomonas aeruginosa and Stenotrophomonas maltophilia strains and complex mixed population biofilms. Experiments using disinfectant and antibiofilm reagents demonstrate that the biofilm sensor is able to discriminate between inactivation/killing of bacteria and destabilization of biofilm structures. The impedance and amperometric sensor data demonstrated the high dynamics of biofilms as a consequence of distinct responses to chemical treatment strategies. Gene expression of flagellar and fimbrial genes of biofilms grown inside the microfluidic system supported the detected biofilm growth kinetics. Thus, the presented biosensor platform is a qualified tool for assessing biofilm formation in specific environments and for evaluating the effectiveness of antibiofilm treatment strategies.

Sub-inhibitory concentrations of antibiotics and wastewater influencing biofilm formation and gene expression of multi-resistant Pseudomonas aeruginosa wastewater isolates.

Sub-inhibitory concentrations of antibiotics, which are found in environmental water systems and sewage plants due to an increased use in therapeutical and preventive fields, influence bacterial behavior in biofilms. The application of sulfamethoxazole, erythromycin, and roxithromycin induced changes in biofilm dynamics regarding biomass formation, spatial structure and specific gene expression in different Pseudomonas aeruginosa isolates. Exposing multi-resistant environmental isolated strains for 17 h to environmental concentrations of antibiotics or wastewater, directly, an increase in biofilm biomass and thickness could be observed for each strain. Additionally, multi-resistant strains responded to the applied growth conditions with changes in transcriptional activity. Here, sub-inhibitory concentrations of macrolides specifically upregulated expression of quorum sensing genes (rhlR, lasI), whereas sulfonamides and municipal wastewater, instead upregulated expression of specific resistant genes (sul1) and efflux pumps (mexD). Antibiotic sensitive isolates demonstrated an overall higher transcriptionally activity, but did not show a specific gene response to the applied exogenous stimuli. Furthermore, the presence of low concentrated antibiotics induced also phenotypical change in the biofilm architecture observed by 3D-imaging.