Scholarly Work - Center for Biofilm Engineering

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    Clothing Textiles as Carriers of Biological Ice Nucleation Active Particles
    (American Chemical Society, 2024-03) Teska, Christy J.; Dieser, Markus; Foreman, Christine M.
    Microplastics have littered the globe, with synthetic fibers being the largest source of atmospheric microplastics. Many atmospheric particles can act as ice nucleators, thereby affecting the microphysical and radiative properties of clouds and, hence, the radiative balance of the Earth. The present study focused on the ice-nucleating ability of fibers from clothing textiles (CTs), which are commonly shed from the normal wear of apparel items. Results from immersion ice nucleation experiments showed that CTs were effective ice nucleators active from −6 to −12 °C, similar to common biological ice nucleators. However, subsequent lysozyme and hydrogen peroxide digestion stripped the ice nucleation properties of CTs, indicating that ice nucleation was biological in origin. Microscopy confirmed the presence of biofilms (i.e., microbial cells attached to a surface and enclosed in an extracellular polysaccharide matrix) on CTs. If present in sufficient quantities in the atmosphere, biological particles (biofilms) attached to fibrous materials could contribute significantly to atmospheric ice nucleation.
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    Anti-Biofilm Efficacy of Commonly Used Wound Care Products in In Vitro Settings
    (MDPI AG, 2023-03) Regulski, Matthew; Myntti, Matthew F.; James, Garth A.
    Considering the prevalence and pathogenicity of biofilms in wounds, this study was designed to evaluate the anti-biofilm capabilities of eight commercially available wound care products using established in vitro assays for biofilms. The products evaluated included dressings with multiple delivery formats for ionic silver including nanocrystalline, gelling fibers, polyurethane (PU) foam, and polymer matrix. Additionally, non-silver-based products including an extracellular polymeric substance (EPS)-dissolving antimicrobial wound gel (BDWG), a collagenase-based debriding ointment and a fish skin-based skin substitute were also evaluated. The products were evaluated on Staphylococcus aureus and Pseudomonas aeruginosa mixed-species biofilms grown using colony drip flow reactor (CDFR) and standard drip flow reactor (DFR) methodologies. Anti-biofilm efficacy was measured by viable plate counts and confocal scanning laser microscopy (CSLM). Four of the eight wound care products tested were efficacious in inhibiting growth of new biofilm when compared with untreated controls. These four products were further evaluated against mature biofilms. BDWG was the only product that achieved greater than 2-log growth reduction (5.88 and 6.58 for S. aureus and P. aeruginosa, respectively) of a mature biofilm. Evaluating both biofilm prevention and mature biofilm disruption capacity is important to a comprehensive understanding of the anti-biofilm efficacy of wound care products.
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    Antimicrobial effects of an acidified nitrite foam on drip flow reactor biofilm
    (European Wound Management Association, 2024-04) Miller, C. Michael; James, Garth; Bell, David; Schultz, Greg
    Background. Nitric oxide (NO) plays critical roles in wound healing, including stimulating vasodilation, angiogenesis and broad antimicrobial activity. Aim. To measure the effect of an acidified nitrite foam (ANF) on biofilms created by six different microbes. Methods. A novel method of generating, delivering and topically applying NO gas at the point of care was developed using ANF in a mixed bubble foam and was tested in vitro against six common microbial wound pathogens. Results. A single 5-minute topical exposure of the NO bubble gas formulation generated a 5.8-log10 reduction of mature biofilm of Pseudomonas aeruginosa, a 5.1-log10 reduction of Staphylococcus aureus biofilm, a 4.0-log10 reduction of Staphylococcus epidermidis biofilm, a 3.2-log10 reduction of Proteus mirabilis biofilm, a 2.7-log10 reduction of Acinetobacter baumannii biofilm, and a 1.5-log10 reduction of Candida albicans biofilm. Conclusion. The efficacy of a 5-minute treatment of ANF used on biofilms of P. aeruginosa, A. baumannii, S. aureus, C. albicans, P. mirabilis and S. epidermidis was confirmed. The treatment resulted in a significant reduction in colony-forming units per square centimetre (CFU/cm2) comparable to or surpassing other methods of NO gas application, suggesting NO containing foam’s utility as a point of care solution for chronic wounds with elevated bioburden and biofilms where levels of endogenously produced NO may be insufficient for wound healing completion.
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    Critical analysis of methods to determine growth, control and analysis of biofilms for potential non-submerged antibiofilm surfaces and coatings
    (Elsevier BV, 2024-06) Redfern, J.; Cunliffe, A. J.; Goeres, D. M.; Azevedo, N. F.; Verran, J.
    The potential uses for antibiofilm surfaces reach across different sectors with significant resultant economic, societal and health impact. For those interested in using antibiofilm surfaces in the built environment, it is important that efficacy testing methods are relevant, reproducible and standardised where possible, to ensure data outputs are applicable to end-use, and comparable across the literature. Using pre-defined keywords, a review of literature reporting on antimicrobial surfaces (78 articles), within which a potential application was described as non-submerged/non-medical surface or coating with antibiofilm action, was undertaken. The most used methods utilized the growth of biofilm in submerged and static systems. Quantification varied (from most to least commonly used) across colony forming unit counts, non-microscopy fluorescence or spectroscopy, microscopy analysis, direct agar-contact, sequencing, and ELISA. Selection of growth media, microbial species, and incubation temperature also varied. In many cases, definitions of biofilm and attempts to quantify antibiofilm activity were absent or vague. Assessing a surface after biofilm recovery or assessing potential regrowth of a biofilm after initial analysis was almost entirely absent. It is clear the field would benefit from widely agreed and adopted approaches or guidance on how to select and incorporate end-use specific conditions, alongside minimum reporting guidelines may benefit the literature.
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    Development of Martian saline seep models and their implications for planetary protection
    (Elsevier BV, 2023-12) Mettler, Madelyn K.; Goemann, Hannah M.; Mueller, Rebecca C.; Vanegas, Oscar A.; Lopez, Gabriela; Singh, Nitin; Venkateswaran, Kasthuri; Peyton, Brent M.
    While life on Mars has not been found, Earth-based microorganisms may contaminate the Red Planet during rover expeditions and human exploration. Due to the survival advantages conferred by the biofilm morphology to microorganisms, such as resistance to UV and osmotic stress, biofilms are particularly concerning from a planetary protection perspective. Modeling and data from the NASA Phoenix mission indicate that temporary liquid water might exist on Mars in the form of high salinity brines. These brines could provide colonization opportunities for terrestrial microorganisms brought by spacecraft or humans. To begin testing for potential establishment of microbes, results are presented from a simplified laboratory model of a Martian saline seep inoculated with sediment from Hailstone Basin, a terrestrial saline seep in Montana (USA). The seep was modeled as a sand-packed drip flow reactor at room temperature fed media with either 1 M MgSO4 or 1 M NaCl. Biofilms were established within the first sampling point of each experiment. Endpoint 16S rRNA gene community analysis showed significant selection of halophilic microorganisms by the media. Additionally, we detected 16S rRNA gene sequences highly similar to microorganisms previously detected in two spacecraft assembly cleanrooms. These experimental models provide an important foundation for identifying microbes that could hitch-hike on spacecraft and may be able to colonize Martian saline seeps. Future model optimization will be vital to informing cleanroom sterilization procedures.
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    Anti-Biofilm Efficacy of Commonly Used Wound Care Products in In Vitro Settings
    (MDPI AG, 2023-03) Regulski, Matthew; Myntti, Matthew F.; James, Garth A.
    Considering the prevalence and pathogenicity of biofilms in wounds, this study was designed to evaluate the anti-biofilm capabilities of eight commercially available wound care products using established in vitro assays for biofilms. The products evaluated included dressings with multiple delivery formats for ionic silver including nanocrystalline, gelling fibers, polyurethane (PU) foam, and polymer matrix. Additionally, non-silver-based products including an extracellular polymeric substance (EPS)-dissolving antimicrobial wound gel (BDWG), a collagenase-based debriding ointment and a fish skin-based skin substitute were also evaluated. The products were evaluated on Staphylococcus aureus and Pseudomonas aeruginosa mixed-species biofilms grown using colony drip flow reactor (CDFR) and standard drip flow reactor (DFR) methodologies. Anti-biofilm efficacy was measured by viable plate counts and confocal scanning laser microscopy (CSLM). Four of the eight wound care products tested were efficacious in inhibiting growth of new biofilm when compared with untreated controls. These four products were further evaluated against mature biofilms. BDWG was the only product that achieved greater than 2-log growth reduction (5.88 and 6.58 for S. aureus and P. aeruginosa, respectively) of a mature biofilm. Evaluating both biofilm prevention and mature biofilm disruption capacity is important to a comprehensive understanding of the anti-biofilm efficacy of wound care products.
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    Antimicrobial Coating Efficacy for Prevention of Pseudomonas aeruginosa Biofilm Growth on ISS Water System Materials
    (Frontiers Media SA, 2022-04) Mettler, Madelyn K.; Parker, Ceth W.; Venkateswaran, Kasthuri; Peyton, Brent M.
    Biofilms can lead to biofouling, microbially induced corrosion, physical impediment and eventual loss in function of water systems, and other engineered systems. The remoteness and closed environment of the International Space Station (ISS) make it vulnerable to unchecked biofilm growth; thus, biofilm mitigation strategies are crucial for current ISS operation and future long duration and deep-space crewed missions. In this study, a space flown bacterial strain of Pseudomonas aeruginosa (PA14) was used as a model organism for its ability to form biofilms. Additionally, a novel antimicrobial coating’s ability to reduce biofilm accumulation on stainless steel, Teflon, titanium, and Inconel (all used in the ISS water treatment and handling systems) was analyzed. Coated materials demonstrated reductions of P. aeruginosa biofilm across all materials when tested in a continuous flow system with tryptic soy broth medium. However, the coating lost efficacy in potato dextrose broth medium. These findings were corroborated via scanning electron microscopy. This study illustrates the fundamental importance of using multiple approaches to test antibiofilm strategies, as well as the specificity in which conditions such strategies can be implemented.
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