Portable Nanofiber-Light Addressable Potentiometric Sensor for Rapid Escherichia coli Detection in Orange Juice.

The growing need to prevent pathogen outbreaks is irrefutable in the case of the food industry. Early detection in products, especially beverages, contaminated with bacterial strains is vital to avoid infected foods from reaching the consumer. If E. coli is pesent in such foods, it can cause infections. It can also be an indicator of the existence of other harmful coliforms. In this study, we have investigated the detection of Escherichia coli ( E. coli) in orange juice using a portable nanofiber-light addressable potentiometric sensor (NF-LAPS). We have chosen electrospun pH-sensitive poly(vinyl alcohol)/poly(acrylic acid) (PVA/PAA) hydrogel NFs as the sensitive layer. The successful detection of E. coli was reported with the NF-LAPS in less than 1 h. The limit of detection (LOD) measured in the sensor is found to be102 CFU/mL. We have confirmed the selectivity of the biosensor toward E. coli by examining the response of the NF-LAPS against Salmonella typhimurium ( S. typhi), also commonly found in orange juice. Despite the complex nature of orange juice, the response of the biosensor is in no way affected while orange juice is tested as is.

Abiotic streamers in a microfluidic system.

In this work, we report the phenomenon of formation of particle aggregates in the form of thin slender strings when a polyacrylamide (PAM) solution, laden with polystyrene (PS) beads is introduced into a microfluidic device containing an array of micropillars. PAM and a dilute solution of PS beads are introduced into the microfluidic channel through two separate inlets and localized particle aggregation is found to occur under certain flow regimes. The particle aggregates initially have a string-like morphology and are tethered at their ends to the micropillar walls, while the structure remains suspended in the fluid medium. Such a morphology inspired us to name these structures streamers. The flow regimes under which streamer formation is observed are quantified through state diagrams. We discuss the streamer formation time-scales and also show that streamer formation is likely the result of the flocculation of PS beads. Streamer formation has implications in investigating particle-laden complex flows through porous media.

Dynamics of bacterial streamers induced clogging in microfluidic devices.

Using a microfabricated porous media mimic platform, we investigated the clogging dynamics of bacterial biomass that accumulated in the device due to the formation of bacterial streamers. Particularly, we found the existence of a distinct clogging front which advanced via pronounced 'stick-slip' of the viscoelastic bacterial biomass over the solid surface of the micro pillar. Thus, the streamer, the solid surface, and the background fluidic media defined a clear three-phase front influencing these advancing dynamics. Interestingly, we also found that once the clogging became substantial, contrary to a static homogenous saturation state, the clogged mimic exhibited an instability phenomena marked by localized streamer breakage and failure leading to extended water channels throughout the mimic. These findings have implications for design and fabrication of biomedical devices and membrane-type systems such as porous balloon catheters, porous stents and filtration membranes prone to bacteria induced clogging as well as understanding bacterial growth and proliferation in natural porous media such as soil and rocks.

Bacterial floc mediated rapid streamer formation in creeping flows.

One of the central puzzles concerning the interaction of low Reynolds number fluid transport with bacterial biomass is the formation of filamentous structures called streamers. In this manuscript, we report our discovery of a new kind of low Re bacterial streamers, which appear from pre-formed bacterial flocs. In sharp contrast to the biofilm-mediated streamers, these streamers form over extremely small timescales (less than a second). Our experiments, carried out in a microchannel with micropillars rely on fluorescence microscopy techniques to illustrate that floc-mediated streamers form when a freely-moving floc adheres to the micropillar wall and gets rapidly sheared by the background flow. We also show that at their inception the deformation of the flocs is dominated by recoverable large strains indicating significant elasticity. These strains subsequently increase tremendously to produce filamentous streamers. Interestingly, we find that these fully formed streamers are not static structures and show viscous response at time scales larger than their formation time scales. Finally, we show that such novel streamer formation can lead to rapid clogging of microfluidic devices.

Protocol for biofilm streamer formation in a microfluidic device with micro-pillars.

Several bacterial species possess the ability to attach to surfaces and colonize them in the form of thin films called biofilms. Biofilms that grow in porous media are relevant to several industrial and environmental processes such as wastewater treatment and CO2 sequestration. We used Pseudomonas fluorescens, a Gram-negative aerobic bacterium, to investigate biofilm formation in a microfluidic device that mimics porous media. The microfluidic device consists of an array of micro-posts, which were fabricated using soft-lithography. Subsequently, biofilm formation in these devices with flow was investigated and we demonstrate the formation of filamentous biofilms known as streamers in our device. The detailed protocols for fabrication and assembly of microfluidic device are provided here along with the bacterial culture protocols. Detailed procedures for experimentation with the microfluidic device are also presented along with representative results.