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In this work, we quantitatively compare computer simulations and existing cell tracking data of P. aeruginosa surface motility in order to analyse the underlying motility mechanism. We present a three dimensional twitching motility model, that simulates the extension, retraction and surface association of individual Type IV Pili (TFP), and is informed by recent experimental observations of TFP. Sensitivity analysis is implemented to minimise the number of model parameters, and quantitative estimates for the remaining parameters are inferred from tracking data by approximate Bayesian computation. We argue that the motility mechanism is highly sensitive to experimental conditions. We predict a TFP retraction speed for the tracking data we study that is in a good agreement with experimental results obtained under very similar conditions. Furthermore, we examine whether estimates for biologically important parameters, whose direct experimental determination is challenging, can be inferred directly from tracking data. One example is the width of the distribution of TFP on the bacteria body. We predict that the TFP are broadly distributed over the bacteria pole in both walking and crawling motility types. Moreover, we identified specific configurations of TFP that lead to transitions between walking and crawling states.
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Biología Computacional , Simulación por Computador , Fimbrias Bacterianas , Modelos Biológicos , Pseudomonas aeruginosa , Pseudomonas aeruginosa/fisiología , Fimbrias Bacterianas/fisiología , Teorema de Bayes , Movimiento/fisiologíaRESUMEN
Polycationic polymers are widely studied antiseptics, and their efficacy is usually quantified by the solution concentration required to kill a fraction of a population of cells (e.g., by Minimum Bactericidal Concentration (MBC)). Here we describe how the response to a polycationic antimicrobial varies greatly among members of even a monoclonal population of bacteria bathed in a single common antimicrobial concentration. We use fluorescence microscopy to measure the adsorption of a labeled cationic polymer, polydiallyldimethylammmonium chloride (PDADMAC, Mw ≈ 4 × 105 g mol-1) and the time course of cell response via a cell permeability indicator for each member of an ensemble of either Escherichia coli, Staphylococcus aureus, or Pseudomonas aeruginosa cells. This is a departure from traditional methods of evaluating synthetic antimicrobials, which typically measure the overall response of a collection of cells at a particular time and therefore do not assess the diversity within a population. Cells typically die after they reach a threshold adsorption of PDADMAC, but not always. There is a substantial time lag of about 5-10 min between adsorption and death, and the time to die of an individual cell is well correlated with the rate of adsorption. The amount adsorbed and the time-to-die differ among species but follow a trend of more adsorption on more negatively charged species, as expected for a cationic polymer. The study of individual cells via time-lapse microscopy reveals additional details that are lost when measuring ensemble properties at a particular time.
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Escherichia coli , Pseudomonas aeruginosa , Staphylococcus aureus , Escherichia coli/efectos de los fármacos , Staphylococcus aureus/efectos de los fármacos , Pseudomonas aeruginosa/efectos de los fármacos , Polietilenos/química , Polietilenos/farmacología , Compuestos de Amonio Cuaternario/farmacología , Compuestos de Amonio Cuaternario/química , Polielectrolitos/química , Polielectrolitos/farmacología , Pruebas de Sensibilidad Microbiana , Antibacterianos/farmacología , Antibacterianos/química , Antibacterianos/síntesis química , Polímeros/farmacología , Polímeros/química , Microscopía Fluorescente , AdsorciónRESUMEN
Some very effective antimicrobial coatings exploit copper or cuprous oxide (Cu2O) as the active agent. The aim of this study is to determine which species is the active antimicrobial - dissolved ions, the Cu2O solid, or reactive oxygen species. Copper ions were leached from Cu2O into various solutions and the leachate tested for both dissolved copper and the efficacy in killing Pseudomonas aeruginosa. The concentration of copper species leached from Cu2O into aqueous solution varied greatly with the composition of the aqueous solution. For a range of solution buffers, killing of P. aeruginosa was highly correlated with the concentration of copper in the leachate. Further, 10 µL bacterial suspension droplets were placed on Cu2O coatings, with or without a polymer barrier layer, and tested for bacterial kill. Killing occurred without contact between bacterium and solid, demonstrating that contact with Cu2O is not necessary. We therefore conclude that soluble copper species are the antimicrobial agent, and that the most potent species is Cu+. The solid quickly raises and sustains the concentration of soluble copper species near the bacterium. Killing via soluble copper ions rather than contact should allow copper coatings to kill bacteria even when fouled, which is an important practical consideration.
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Antiinfecciosos , Cobre , Cobre/farmacología , Antibacterianos/farmacología , Antiinfecciosos/farmacología , Bacterias , Pseudomonas aeruginosa , IonesRESUMEN
Evaporation of water using solar power is an economical and environmentally friendly method for purification of aqueous solutions. It has been suggested that intermediate states can be used to lower the enthalpy of evaporation of water and therefore to increase the efficiency of evaporation that uses absorption of sunlight. However, the relevant quantity is the enthalpy of evaporation from bulk water to bulk vapor, which is fixed for a given temperature and pressure. The formation of an intermediate state does not alter the enthalpy of the overall process.
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Antimicrobial coatings have a finite lifetime because of wear, depletion of the active ingredient, or surface contamination that produces a barrier between the pathogen and the active ingredient. The limited lifetime means that facile replacement is important. Here, we describe a generic method for rapidly applying and reapplying antimicrobial coatings to common-touch surfaces. The method is to deposit an antimicrobial coating on a generic adhesive film (wrap), and then to attach that modified wrap to the common-touch surface. In this scenario, the adhesion of the wrap and antimicrobial efficacy are separated and can be optimized independently. We demonstrate the fabrication of two antimicrobial wraps, both using cuprous oxide (Cu2O) as the active ingredient. The first uses polyurethane (PU) as the polymeric binder and the second uses polydopamine (PDA). Our antimicrobial PU/Cu2O and PDA/Cu2O wraps, respectively, kill >99.98% and >99.82% of the human pathogen, P. aeruginosa, in only 10 min, and each of them kill >99.99% of the bacterium in 20 min. These antimicrobial wraps can be removed and replaced on the same object in <1 min with no tools. Wraps are already frequently used by consumers to coat drawers or cars for aesthetic or protective purposes.
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Antimicrobial coatings can be used to reduce the transmission of infectious agents that are spread by contact. An effective coating should kill microbes in the time between users, which is sometimes minutes or less. Fast killing requires fast transport, and our proposed method of fast transport is a porous coating where the contaminated liquid imbibes (infiltrates) into the pores to achieve rapid contact with active material inside the pores. We test the hypothesis that a porous antimicrobial coating will enable faster inactivation of microorganisms than a planar coating of the same material. We use hydrophilic pores with dimensions of 5-100 µm such that liquid droplets imbibe in seconds, and from there transport distances and times are short, defined by the pore size rather than the droplet size. Our coating has two levels of structure: (A) a porous scaffold and (B) an antimicrobial coating within the pore structure containing the active ingredient. Two scaffolds are studied: stainless steel and poly(methyl methacrylate) (PMMA). The active ingredient is electrolessly deposited copper. To enhance adhesion and growth of copper, a layer of polydopamine (PDA) is deposited on the scaffold prior to deposition of the copper. This porous copper coating kills 99.84% of Pseudomonas aeruginosa within 3 min, which is equivalent to a half-life of 27 s. In contrast, the same layer of PDA/copper on a nonporous coating kills 79.65% in the same time frame, consistent with the hypothesis that the killing rate is increased by the addition of porosity. Using the porous PMMA scaffold, the porous antimicrobial coating kills >99.99% P. aeruginosa in 5 min, which is equivalent to a half-life of 21 s. The higher rate of kill on the porous antimicrobial solid is appropriate for hindering the spread of infectious agents on common-use objects.
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Antiinfecciosos , Polimetil Metacrilato , Porosidad , Cobre/química , Antibacterianos/farmacología , Antibacterianos/química , Antiinfecciosos/farmacologíaRESUMEN
HYPOTHESIS: The addition of a thin, hydrophilic, porous, coating to an impermeable solid will lead to more rapid evaporation of liquid droplets that impinge on the solid. The droplet will imbibe quickly, but the progress normal to the interface will be limited to the thickness of the coating, and therefore the liquid will spread laterally into a broad disk to expose a large liquid-vapor interface for evaporation. EXPERIMENTS: Liquid droplets of volume 2.5-25 µL were placed on solids and then both the mass and area of each droplet were monitored over time. We compared data for smooth, impermeable hydrophilic glass to the same glass that was coated in thin (35-109 µm) porous, hydrophilic-glass layer fabricated from glass beads. FINDINGS: The droplet was imbibed (wicked) into the coating within seconds, and the liquid spread laterally to form a thin, broad, disk. Critically, evaporation of a droplet was enhanced by a factor of 7-8 on the thin coating. The evaporation rate was not proportional to the reciprocal thickness of the coating. The ability to enhance evaporation of small droplets on a solid may have practical applications, for example, in speeding the death of microbes.
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Gases , Porosidad , Interacciones Hidrofóbicas e HidrofílicasRESUMEN
Antimicrobial coatings can inhibit the transmission of infectious diseases when they provide a quick kill that is achieved long after the coating application. Here, we describe the fabrication and testing of a glass coating containing Ag2O microparticles that was prepared from sodium silicate at room temperature. The half-lives of both methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa on this coating are only 2-4 min. The half-life of Clostridioides difficile spores is about 9-12 min, which is extremely short for a spore. Additional tests on MRSA demonstrate that the coating retains its antimicrobial activity after abrasion and that an increased loading of Ag2O leads to a shorter half-life. This coating combines the properties of optical transparency, robustness, fast kill, and room temperature preparation that are highly desirable for an antimicrobial coating.
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This study investigates how the recent history of bacteria affects their attachment to a solid-liquid interface. We compare the attachment from a flowing suspension of the bacterium,Pseudomonas aeruginosaPAO1, after one of two histories: (a) passage through a tube packed with glass beads or (b) passage through an empty tube. The glass beads were designed to increase the rate of bacterial interactions with solid-liquid surfaces prior to observation in a flow cell. Analysis of time-lapse microscopy of the bacteria in the flow cells shows that the residence time distribution and surface density of bacteria differ for these two histories. In particular, bacteria exiting the bead-filled tube, in contrast to those bacteria exiting the empty tube, are less likely to attach to the subsequent flow cell window and begin surface growth. In contrast, when we compared two histories defined by different lengths of tubing, there was no difference in either the mean residence time or the surface density. In order to provide a framework for understanding these results, we present a phenomenological model in which the rate of bacterial surface density growth,dN(t)/dt, depends on two terms. One term models the initial attachment of bacteria to a surface, and is proportional to the nonprocessive cumulative residence time distribution for bacteria that attach and detach from the surface without cell division. The second term for the rate is proportional to the bacterial surface density and models surface cell division. The model is in surprisingly good agreement with the data even though the surface growth process is a complex interplay between attachment/detachment at the solid-liquid interface and cell division on the surface.
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Adhesión Bacteriana , Biopelículas , Pseudomonas aeruginosa , Bacterias , Propiedades de SuperficieRESUMEN
Pathogenic bacteria can remain viable on fabrics for several days and therefore are a source of infection. Antimicrobial fabrics are a potential method of reducing such infections, and advances in antimicrobial fabrics can be enhanced by knowledge of how the fabric kills bacteria. Metal oxides have been considered and used as antimicrobial ingredients in self-sanitizing surfaces, including in clinical settings. In this work, we examine how the addition of cuprous oxide (Cu2O) particles to polypropylene fibers kills bacteria. First, we show that the addition of the Cu2O particles reduces the viability of common hospital pathogens, Staphylococcus aureus, Pseudomonas aeruginosa, and Streptococcus pneumoniae, by 99.9% after 30 min of contact with the treated polypropylene. Then, we demonstrate that the main killing effect is due to the drying of the bacteria onto the cuprous oxide particles. There is also a weaker effect due to free Cu+ ions that dissolve into the liquid. Other dissolved species were unimportant. Chelation of these Cu+ ions in soluble form or precipitation removes their antimicrobial activity.
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Previous reports indicated the low stability of severe actute respiratory syndrome coronovirus 2 (SARS-CoV-2) on various porous surfaces, but the role of porosity was unclear because there was no direct comparison between porous and nonporous solids of the same chemistry. Through comparing pairs of solids with very similar chemistry, we find that porosity is important: porous glass has a much lower infectivity than nonporous glass. However, porosity is not sufficient to lower infectivity; permeability, which is the ability of a liquid to move through a material, is the important parameter. We show this by comparing a pair of porous CuO coatings where the pores are accessible in one case and inaccessible in the other case. When the pores are inaccessible, the infectivity remains similar to that for nonporous solids. Thus, for both glass and CuO, it is the access to porosity that decreases the infectivity of extracted liquid droplets. Having established the importance of permeability, there is the open question of the mechanism of changing the infectivity of SARS-CoV-2. Several hypotheses are possible, such as increasing the difficulty of extracting the virus from the solid, changing the drying time, increasing the surface area of active ingredient, etc. Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) measurements show that less viral DNA is extracted from a permeable surface, suggesting that the virus becomes trapped in the pores. Finally, we consider the effect of drying. We show that permeability and the water contact angle on the solid have effects on the drying time of a contaminated droplet, which may in turn affect infectivity.
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We measured the diffusion of an anion, fluorescein, confined to a nanoscale (10-100 nm) aqueous film between two glass walls. The two glass walls were very slightly angled to form a crack. The diffusion of fluorescein was strongly influenced by the presence of an inert electrolyte, NaCl, in the film prior to the diffusion of charged fluorescein into the crack. The time to reach an equilibrium distribution of fluorescein was 10 times longer without the inert electrolyte than when the electrolyte was present. In applications where rapid diffusion of ions is important, it would therefore be advisable to not prewet a confined space with pure water. We attribute this phenomenon to the effect of the electrical potential of the confining walls. Unscreened surface potential in a thin film severely hinders the diffusion of the fluorescein ion. As salt diffuses into the thin film, the electrostatic double layer shrinks in thickness and further diffusion of ions is less hindered. On the other hand, diffusion of ions into the film is only weakly affected by the Debye length of the solution, provided that the surface potential inside a thin film is initially screened by even a low concentration of electrolyte inside the film. The time evolution of the concentration profile for different Debye lengths matches a diffusion model developed with the finite difference method (FDM).
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Transparent antimicrobial coatings can maintain the aesthetic appeal of surfaces and the functionality of a touch-screen while adding the benefit of reducing disease transmission. We fabricated an antimicrobial coating of silver oxide particles in a silicate matrix on glass. The matrix was grown by a modified Stöber sol-gel process with vapor-phase water and ammonia. A coating on glass with 2.4 mg of Ag2O per mm2 caused a reduction of 99.3% of SARS-CoV-2 and >99.5% of Pseudomonas aeruginosa, Staphylococcus aureus, and methicillin-resistant Staphylococcus aureus compared to the uncoated glass after 1 h. We envisage that screen protectors with transparent antimicrobial coatings will find particular application to communal touch-screens, such as in supermarkets and other check-out or check-in facilities where a number of individuals utilize the same touch-screen in a short interval.
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Antiinfecciosos/química , Infecciones Bacterianas/prevención & control , COVID-19/prevención & control , Óxidos/química , Compuestos de Plata/química , Amoníaco/química , Antiinfecciosos/farmacología , Infecciones Bacterianas/microbiología , COVID-19/virología , Vidrio/química , Humanos , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Staphylococcus aureus Resistente a Meticilina/patogenicidad , Óxidos/farmacología , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/patogenicidad , SARS-CoV-2/efectos de los fármacos , SARS-CoV-2/patogenicidad , Silicatos/química , Compuestos de Plata/farmacología , Agua/químicaRESUMEN
Transfer of SARS-CoV-2 from solids to fingers is one step in infection via contaminated solids, and the possibility of infection from this route has driven calls for increased frequency of handwashing during the COVID-19 pandemic. To analyze this route of infection, we measured the percentage of SARS-CoV-2 that was transferred from a solid to an artificial finger. A droplet of SARS-CoV-2 suspension (1 µL) was placed on a solid, and then artificial skin was briefly pressed against the solid with a light force (3 N). Transfer from a variety of solids was detected, and transfer from the non-porous solids, glass, stainless steel, and Teflon, was substantial when the droplet was still wet. The viral titer for the finger was 13-16% or 0.8-0.9 log less than for the input droplet. Transfer still occurred after the droplet evaporated, but was smaller, 3-9%. We found a lower level of transfer from porous solids but did not find a significant effect of solid wettability for non-porous solids.
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COVID-19/transmisión , Transmisión de Enfermedad Infecciosa/prevención & control , SARS-CoV-2/metabolismo , COVID-19/metabolismo , Contaminación de Equipos/prevención & control , Contaminación de Equipos/estadística & datos numéricos , Humanos , SARS-CoV-2/patogenicidad , Piel/virología , Carga ViralRESUMEN
Antimicrobial coatings are one method to reduce the spread of microbial diseases. Transparent coatings preserve the visual properties of surfaces and are strictly necessary for applications such as antimicrobial cell phone screens. This work describes transparent coatings that inactivate microbes within minutes. The coatings are based on a polydopamine (PDA) adhesive, which has the useful property that the monomer can be sprayed, and then the monomer polymerizes in a conformal film at room temperature. Two coatings are described (1) a coating where PDA is deposited first and then a thin layer of copper is grown on the PDA by electroless deposition (PDA/Cu) and (2) a coating where a suspension of Cu2O particles in a PDA solution is deposited in a single step (PDA/Cu2O). In the second coating, PDA menisci bind Cu2O particles to the solid surface. Both coatings are transparent and are highly efficient in inactivating microbes. PDA/Cu kills >99.99% of Pseudomonas aeruginosa and 99.18% of methicillin-resistant Staphylococcus aureus (MRSA) in only 10 min and inactivates 99.98% of SARS-CoV-2 virus in 1 h. PDA/Cu2O kills 99.94% of P. aeruginosa and 96.82% of MRSA within 10 min and inactivates 99.88% of SARS-CoV-2 in 1 h.
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Antibacterianos/farmacología , Antivirales/farmacología , Farmacorresistencia Microbiana/efectos de los fármacos , SARS-CoV-2/efectos de los fármacos , COVID-19/virología , Humanos , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Pseudomonas aeruginosa/efectos de los fármacos , Propiedades de SuperficieRESUMEN
We developed antimicrobial coatings from ZnO particles that reduce the infectivity of SARS-CoV-2 suspensions by >99.9% in 1 h. The advantage of a coating is that it can be applied to a variety of objects, e.g., hand rails and door knobs, to hinder the spread of disease. Two porous coatings were prepared: one from submicrometer zinc oxide particles bound with silica menisci and the other from zinc oxide tetrapods bound with polyurethane. Experiments on glass coatings show that infectivity depends on porosity for hydrophilic materials, wherein aqueous droplets are imbibed into the pores.
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COVID-19 , Óxido de Zinc , Antibacterianos , Humanos , SARS-CoV-2 , Suspensiones , Óxido de Zinc/farmacologíaRESUMEN
The COVID-19 pandemic had a major impact on life in 2020 and 2021. One method of transmission occurs when the causative virus, SARS-CoV-2, contaminates solids. Understanding and controlling the interaction with solids is thus potentially important for limiting the spread of the disease. We review work that describes the prevalence of the virus on common objects, the longevity of the virus on solids, and surface coatings that are designed to inactivate the virus. Engineered coatings have already succeeded in producing a large reduction in viral infectivity from surfaces. We also review work describing inactivation on facemasks and clothing and discuss probable mechanisms of inactivation of the virus at surfaces.
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The ongoing COVID-19 pandemic has created a need for coatings that reduce infection from SARS-CoV-2 via surfaces. Such a coating could be used on common touch surfaces (e.g., door handles and railings) to reduce both disease transmission and fear of touching objects. Herein, we describe the design, fabrication, and testing of a cupric oxide anti-SARS-CoV-2 coating. Rapid loss of infectivity is an important design criterion, so a porous hydrophilic coating was created to allow rapid infiltration of aqueous solutions into the coating where diffusion distances to the cupric oxide surface are short and the surface area is large. The coating was deposited onto glass from a dispersion of cuprous oxide in ethanol and then thermally treated at 700 °C for 2 h to produce a CuO coating that is ≈30 µm thick. The heat treatment oxidized the cuprous oxide to cupric oxide and sintered the particles into a robust film. The SARS-CoV-2 infectivity from the CuO film was reduced by 99.8% in 30 min and 99.9% in 1 h compared to that from glass. The coating remained hydrophilic for at least 5 months, and there was no significant change in the cross-hatch test of robustness after exposure to 70% ethanol or 3 wt % bleach.
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COVID-19/prevención & control , COVID-19/virología , Cobre/farmacología , SARS-CoV-2/fisiología , Animales , Chlorocebus aethiops , Humanos , Espectroscopía de Fotoelectrones , SARS-CoV-2/efectos de los fármacos , Propiedades de Superficie , Células Vero , Inactivación de Virus/efectos de los fármacos , Difracción de Rayos XRESUMEN
SARS-CoV-2, the virus that causes the disease COVID-19, remains viable on solids for periods of up to 1 week, so one potential route for human infection is via exposure to an infectious dose from a solid. We have fabricated and tested a coating that is designed to reduce the longevity of SARS-CoV-2 on solids. The coating consists of cuprous oxide (Cu2O) particles bound with polyurethane. After 1 h on coated glass or stainless steel, the viral titer was reduced by about 99.9% on average compared to the uncoated sample. An advantage of a polyurethane-based coating is that polyurethane is already used to coat a large number of everyday objects. Our coating adheres well to glass and stainless steel as well as everyday items that people may fear to touch during a pandemic, such as a doorknob, a pen, and a credit card keypad button. The coating performs well in the cross-hatch durability test and remains intact and active after 13 days of being immersed in water or after exposure to multiple cycles of exposure to the virus and disinfection.
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Betacoronavirus/genética , Infecciones por Coronavirus/genética , Pandemias , Neumonía Viral/genética , Betacoronavirus/patogenicidad , COVID-19 , Cobre/química , Cobre/metabolismo , Infecciones por Coronavirus/virología , Humanos , Neumonía Viral/virología , Poliuretanos/química , Poliuretanos/metabolismo , SARS-CoV-2 , Propiedades de SuperficieRESUMEN
Air bubbles are a promising means of controlling fouling for a range of applications, particularly delaying fouling in marine environments. This work investigates the mechanism by which the collision of an air bubble with a solid removes adsorbed bacteria. A key feature of the work is that the numbers of bacteria were monitored via video microscopy throughout the collision; so, we were able to explore the mechanism of bacteria removal. When a bubble collides with a solid, an air-liquid interface crosses the solid twice, and we were able to distinguish the effects of the first and second air-liquid interfaces. The bacterium Pseudomonas aeruginosa was allowed to adhere to smooth poly(dimethylsiloxane) and then a collision with a bubble was investigated for one of three different approach geometries: perpendicular, parallel, and oscillating parallel to the solid surface. Other factors examined were the speed of the bubble, the duration of bacterial adhesion on the solid surface, and the wettability of the solid. Surface wettability was identified as the most significant factor. When the solid dewet, almost all bacteria were removed from hydrophobic surfaces upon the passage of the first air-liquid interface. In contrast, when a thin liquid film remained between the solid and the bubble (a hydrophilic solid), variable amounts of bacteria remained. Although almost all bacteria were initially removed from hydrophobic solids, many bacteria were redeposited on hydrophobic surfaces upon the passage of the second air-liquid interface, especially when the first and second air-liquid interfaces moved in opposite directions. As described previously, a lower velocity of the bubble allows more time for the thin liquid film to drain and improved removal efficiency on hydrophilic solids. A rougher solid (8 µm diameter hemispherical protrusions) decreased the detachment efficiency because bacteria and liquid were able to shelter in concavities.