No abstract

Biofilms in drinking water distribution systems (DWDS) could exacerbate the persistence and associated risks of pathogenic Legionella pneumophila (L. pneumophila), thus raising human health concerns. However, mechanisms controlling adhesion and subsequent detachment of L. pneumophila associated with biofilms remain unclear. We determined the connection between L. pneumophila adhesion and subsequent detachment with biofilm physical structure characterization using optical coherence tomography (OCT) imaging technique. Analysis of the OCT images of multispecies biofilms grown under low nutrient condition up to 34 weeks revealed the lack of biofilm deformation even when these biofilms were exposed to flow velocity of 0.7 m/s, typical flow for DWDS. L. pneumophila adhesion on these biofilm under low flow velocity (0.007 m/s) positively correlated with biofilm roughness due to enlarged biofilm surface area and local flow conditions created by roughness asperities. The preadhered L. pneumophila on selected rough and smooth biofilms were found to detach when these biofilms were subjected to higher flow velocity. At the flow velocity of 0.1 and 0.3 m/s, the ratio of detached cell from the smooth biofilm surface was from 1.3 to 1.4 times higher than that from the rough biofilm surface, presumably because of the low shear stress zones near roughness asperities. This study determined that physical structure and local hydrodynamics control adhesion and detachment from simulated drinking water biofilm, thus it is the first step toward reducing the risk of L. pneumophila exposure and subsequent infections.

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The significance of extracellular polymer substances (EPS) on cell deposition on silica surfaces was examined by direct comparison of the deposition kinetics of untreated "intact" bacteria versus those from the same strain but with EPS removal via cation exchange resin (CER) treatment using a quartz crystal microbalance with dissipation (QCM-D). Four bacterial strains, mutant Escherichia coli BL21 (gram-negative, nonmotile), Pseudomonas sp QG6 (gram-negative, motile), Rhodococcus sp QL2 (gram-positive, nonmotile), and Bacillus subtilis (gram-positive, motile), were employed to determine the influence of EPS on cell deposition. Experiments were conducted in both monovalent (NaCl) and divalent (CaCl2) solutions under a variety of environmentally relevant ionic strength ranging from 1 to 100 mM at pH 6.0. The effectiveness of EPS removal via CER method was ensured by biochemical composition analysis of EPS solutions and further confirmed by FTIR analysis. Comparable zeta potentials were observed for untreated and CER treated bacterial cells in both NaCl and CaCl2 solutions, indicating that removal of EPS from cell surfaces via CER treatment did not affect the electrokinetic properties of the cell surfaces for all four strains. However, observed deposition efficiencies (alpha) were greater for untreated cells relative to those with CER treated cells across the entire ionic strength range examined in both NaCI and CaCl2 solutions for all four bacterial strains. These results strongly demonstrated that the removal of EPS from cell surfaces for all four strains decreased the deposition of bacteria on silica surfaces. This study clearly showed that the enhancement of cell deposition on silica surfaces due to the presence of EPS on cell surfaces was relevant to all bacterial strains examined regardless of cell types and motility.

Mechanical and structural properties of biofilms influence the accumulation and release of pathogens in drinking water distribution systems (DWDS). Thus, understanding how long-term residual disinfectants exposure affects biofilm mechanical and structural properties is a necessary aspect for pathogen risk assessment and control. In this study, elastic modulus and structure of groundwater biofilms was monitored by atomic force microscopy (AFM) and optical coherence tomography (OCT) during three months of exposure to monochloramine or free chlorine. After the first month of disinfectant exposure, the mean stiffness of monochloramine- or free-chlorine-treated biofilms was 4 to 9 times higher than those before treatment. Meanwhile, the biofilm thickness decreased from 120 ± 8 µm to 93 ± 6–107 ± 11 µm. The increased surface stiffness and decreased biofilm thickness within the first month of disinfectant exposure was presumably due to the consumption of biomass. However, by the second to third month during disinfectant exposure, the biofilm mean stiffness showed a 2- to 4-fold decrease, and the biofilm thickness increased to 110 ± 7–129 ± 8 µm suggesting that the biofilms adapted to disinfectant exposure. After three months of the disinfectant exposure process, the disinfected biofilms showed 2–5 times higher mean stiffness (as determined by AFM) and 6–13-fold higher ratios of protein over polysaccharide, as determined by differential staining and confocal laser scanning microscopy (CLSM), than the nondisinfected groundwater biofilms. However, the disinfected biofilms and nondisinfected biofilms showed statistically similar thicknesses (t test, p > 0.05), suggesting that long-term disinfection may not significantly remove net biomass. This study showed how biofilm mechanical and structural properties vary in response to a complex DWDS environment, which will contribute to further research on the risk assessment and control of biofilm-associated-pathogens in DWDS.

No abstract

Airborne transmission of SARS-CoV-2 plays a critical role in spreading COVID-19. To protect public health, we designed and fabricated electrospun nanofibrous air filters that hold promise for applications in personal protective equipment (PPE) and the indoor environment. Due to ultrafine nanofibers (∼300 nm), the electrospun air filters had a much smaller pore size in comparison to the surgical mask and cloth masks (a couple of micrometers versus tens to hundreds of micrometers). A coronavirus strain served as a SARS-CoV-2 surrogate and was used to generate aerosols for filtration efficiency tests, which can better represent SARS-CoV-2 in comparison to other agents used for aerosol generation in previous studies. The electrospun air filters showed excellent performance by capturing up to 99.9% of coronavirus aerosols, which outperformed many commercial face masks. In addition, we observed that the same electrospun air filter or face mask removed NaCl aerosols equivalently or less effectively in comparison to the coronavirus aerosols when both aerosols were generated from the same system. Our work paves a new avenue for advancing air filtration by developing electrospun nanofibrous air filters for controlling SARS-CoV-2 airborne transmission.

Interactions between bacteriophages (phages) and biofilms remain poorly understood despite the broad implications for microbial ecology, water quality, and microbiome engineering. Here, we demonstrate that lytic coliphage PHH01 can hitchhike on carrier bacteria Bacillus cereus to facilitate its infection of host bacteria, Escherichia coli, in biofilms. Specifically, PHH01 could adsorb onto the flagella of B. cereus, and thus phage motility was increased, resulting in 4.36-fold more effective infection of E. coli in biofilm relative to free PHH01 alone. Moreover, phage infection mitigated interspecies competition and enhanced B. cereus colonization; the fraction of B. cereus in the final biofilm increased from 9% without phages to 43% with phages. The mutualistic relationship between the coliphage and carrier bacteria was substantiated by migration tests on an E. coli lawn: the conjugation of PHH01 and B. cereus enhanced B. cereus colonization by 6.54-fold compared to B. cereus alone (6.15 vs 0.94 cm 2 in 24 h) and PHH01 migration by 5.15-fold compared to PHH01 alone (10.3 vs 2.0 mm in 24 h). Metagenomic and electron microscopic analysis revealed that the phages of diverse taxonomies and different morphologies could be adsorbed by the flagella of B. cereus, suggesting hitchhiking on flagellated bacteria might be a widespread strategy in aquatic phage populations. Overall, our study highlights that hitchhiking behavior in phages can facilitate phage infection of biofilm bacteria, promote carrier bacteria colonization, and thus significantly influence biofilm composition, which holds promise for mediating biofilm functions and moderating associated risks.