Browsing by Author "Parker, Albert E."

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Accelerated Gibbs sampling of normal distributions using matrix splittings and polynomials
(2017-11) Fox, Colin; Parker, Albert E.
Standard Gibbs sampling applied to a multivariate normal distribution with a specified precision matrix is equivalent in fundamental ways to the Gauss-Seidel iterative solution of linear equations in the precision matrix. Specifically, the iteration operators, the conditions under which convergence occurs, and geometric convergence factors (and rates) are identical. These results hold for arbitrary matrix splittings from classical iterative methods in numerical linear algebra giving easy access to mature results in that field, including existing convergence results for antithetic-variable Gibbs sampling, REGS sampling, and generalizations. Hence, efficient deterministic stationary relaxation schemes lead to efficient generalizations of Gibbs sampling. The technique of polynomial acceleration that significantly improves the convergence rate of an iterative solver derived from a symmetric matrix splitting may be applied to accelerate the equivalent generalized Gibbs sampler. Identicality of error polynomials guarantees convergence of the inhomogeneous Markov chain, while equality of convergence factors ensures that the optimal solver leads to the optimal sampler. Numerical examples are presented, including a Chebyshev accelerated SSOR Gibbs sampler applied to a stylized demonstration of low-level Bayesian image reconstruction in a large 3-dimensional linear inverse problem.
Accelerated Gibbs sampling of normal distributions using matrix splittings and polynomials
(2017-11) Fox, Colin; Parker, Albert E.
Standard Gibbs sampling applied to a multivariate normal distribution with a specified precision matrix is equivalent in fundamental ways to the Gauss Seidel iterative solution of linear equations in the precision matrix. Specifically, the iteration operators, the conditions under which convergence occurs, and geometric convergence factors (and rates) are identical. These results hold for arbitrary matrix splittings from classical iterative methods in numerical linear algebra giving easy access to mature results in that field, including existing convergence results for antithetic-variable Gibbs sampling, REGS sampling, and generalizations. Hence, efficient deterministic stationary relaxation schemes lead to efficient generalizations of Gibbs sampling. The technique of polynomial acceleration that significantly improves the convergence rate of an iterative solver derived from a symmetric matrix splitting may be applied to accelerate the equivalent generalized Gibbs sampler. Identicality of error polynomials guarantees convergence of the inhomogeneous Markov chain, while equality of convergence factors ensures that the optimal solver leads to the optimal sampler. Numerical examples are presented, including a Chebyshev accelerated SSOR Gibbs sampler applied to a stylized demonstration of low-level Bayesian image reconstruction in a large 3-dimensional linear inverse problem.
Activity-based cell sorting reveals responses of uncultured archaea and bacteria to substrate amendment
(Springer Science and Business Media LLC, 2020) Reichart, Nicholas J.; Jay, Zachary J.; Krukenberg, Viola; Parker, Albert E.; Lange Spietz, Rachel K.; Hatzenpichler, Roland
Metagenomic studies have revolutionized our understanding of the metabolic potential of uncultured microorganisms in various ecosystems. However, many of these genomic predictions have yet to be experimentally tested, and the functional expression of genomic potential often remains unaddressed. In order to obtain a more thorough understanding of cell physiology, novel techniques capable of testing microbial metabolism under close to in situ conditions must be developed. Here, we provide a benchmark study to demonstrate that bioorthogonal non-canonical amino acid tagging (BONCAT) in combination with fluorescence-activated cell sorting (FACS) and 16S rRNA gene sequencing can be used to identify anabolically active members of a microbial community incubated in the presence of various growth substrates or under changing physicochemical conditions. We applied this approach to a hot spring sediment microbiome from Yellowstone National Park (Wyoming, USA) and identified several microbes that changed their activity levels in response to substrate addition, including uncultured members of the phyla Thaumarchaeota, Acidobacteria, and Fervidibacteria. Because shifts in activity in response to substrate amendment or headspace changes are indicative of microbial preferences for particular growth conditions, results from this and future BONCAT-FACS studies could inform the development of cultivation media to specifically enrich uncultured microbes. Most importantly, BONCAT-FACS is capable of providing information on the physiology of uncultured organisms at as close to in situ conditions as experimentally possible.
Activity-based cell sorting reveals responses of uncultured archaea and bacteria to substrate amendment
(2020) Reichart, Nicholas J.; Jay, Zackary J.; Krukenberg, Viola; Parker, Albert E.; Lange Spietz, Rachel K.; Hatzenpichler, Roland
Metagenomic studies have revolutionized our understanding of the metabolic potential of uncultured microorganisms in various ecosystems. However, many of these genomic predictions have yet to be experimentally tested, and the functional expression of genomic potential often remains unaddressed. In order to obtain a more thorough understanding of cell physiology, novel techniques capable of testing microbial metabolism under close to in situ conditions must be developed. Here, we provide a benchmark study to demonstrate that bioorthogonal non-canonical amino acid tagging (BONCAT) in combination with fluorescence-activated cell sorting (FACS) and 16S rRNA gene sequencing can be used to identify anabolically active members of a microbial community incubated in the presence of various growth substrates or under changing physicochemical conditions. We applied this approach to a hot spring sediment microbiome from Yellowstone National Park (Wyoming, USA) and identified several microbes that changed their activity levels in response to substrate addition, including uncultured members of the phyla Thaumarchaeota, Acidobacteria, and Fervidibacteria. Because shifts in activity in response to substrate amendment or headspace changes are indicative of microbial preferences for particular growth conditions, results from this and future BONCAT-FACS studies could inform the development of cultivation media to specifically enrich uncultured microbes. Most importantly, BONCAT-FACS is capable of providing information on the physiology of uncultured organisms at as close to in situ conditions as experimentally possible.
Antimicrobial penetration and efficacy in an in vitro oral biofilm model
(2011-05) Corbin, A.; Pitts, Betsey; Parker, Albert E.; Stewart, Philip S.
The penetration and overall efficacy of six mouthrinse actives was evaluated by using an in vitro flow cell oral biofilm model. The technique involved preloading biofilm cells with a green fluorescent dye that leaked out as the cells were permeabilized by a treatment. The loss of green color, and of biomass, was observed by time-lapse microscopy during 60 min of treatment under continuous flow conditions. The six actives analyzed were ethanol, sodium lauryl sulfate, triclosan, chlorhexidine digluconate (CHX), cetylpyridinium chloride, and nisin. Each of these agents effected loss of green fluorescence throughout biofilm cell clusters, with faster action at the edge of a cell cluster and slower action in the cluster center. The time to reach half of the initial fluorescent intensity at the center of a cell cluster, which can be viewed as a combined penetration and biological action time, ranged from 0.6 to 19 min for the various agents. These times are much longer than the predicted penetration time based on diffusion alone, suggesting that anti-biofilm action was controlled more by the biological action time than by the penetration time of the active. None of the agents tested caused any removal of the biofilm. The extent of fluorescence loss after 1 h of exposure to an active ranged from 87 to 99.5%, with CHX being the most effective. The extent of fluorescence loss in vitro, but not penetration and action time, correlated well with the relative efficacy data from published clinical trials.
Attraction, Entrance, and Passage Efficiency of Arctic Grayling, Trout, and Suckers at Denil Fishways in the Big Hole River Basin, Montana
(Wiley, 2022-07) Triano, Ben; Kappenman, Kevin M.; McMahon, Thomas E.; Blank, Matt; Heim, Kurt C.; Parker, Albert E.; Zale, Alexander V.; Platt, Nolan; Plymesser, Katey
The Big Hole River basin in southwestern Montana supports the only indigenous, self-sustaining fluvial population of Arctic Grayling Thymallus arcticus in the conterminous United States, but the basin is fragmented by numerous low-head irrigation diversion dams. Denil fishways at 63 diversion dams provide Arctic Grayling and other fishes opportunities for year-round access to critical habitats; however, their efficiency has not been evaluated. We quantified attraction, entrance, and passage for hatchery-reared Arctic Grayling, wild trout (Brook Trout Salvelinus fontinalis and Brown Trout Salmo trutta), and wild suckers (White Sucker Catostomus commersonii and Longnose Sucker C. catostomus) during 14 field trials conducted at six Denil fishways over a representative range of fishway slopes and hydraulic conditions using passive integrated transponder telemetry. Attraction (60.4–84.3%) and entrance (44.3–78.6%) efficiencies were variable across test conditions and reduced overall fishway efficiencies (19.1–55.8%). In contrast, upon entry, passage efficiencies were high (96.2–97.0%) for all taxa across all test conditions. Attraction of hatchery-reared Arctic Grayling increased with upstream depth (a surrogate for fishway discharge) and attraction flow, but attraction of wild fish was less affected by these conditions. Entrance of Arctic Grayling, Brook Trout, and Brown Trout decreased with upstream depth and fishway slope, especially when plunging entrance conditions associated with shallow downstream depths were present. However, entrance of Arctic Grayling and both trout species increased with downstream depth, and submerged fishway entrances demonstrated promise for increasing entrance efficiency at fishways with high discharges and steep slopes. We demonstrate that comprehensive evaluations of fishway efficiency components can identify specific solutions that improve fishway efficiency; application of these engineering solutions at individual fishways (as needed) could improve their efficiency and further enhance aquatic connectivity for fishes in the Big Hole River basin and elsewhere.
Bacterial Adhesion and Biofilm Formation on Textured Breast Implant Shell Materials
(2019-04) James, Garth A.; Boegli, Laura; Hancock, John; Bowersock, Lisa B.; Parker, Albert E.; Kinney, Brian M.
"Background Bacterial biofilms have been implicated with breast implant complications including capsular contracture and anaplastic large-cell lymphoma. The actual mechanisms for either are still under active investigation and are not clear. Due to their increased surface area, implants with textured surfaces may harbor greater biofilm loads than those with smooth surfaces. Methods Biofilm formation on the outer surface material was compared using implants with various surface areas and roughness, including Natrelle® (Smooth), SmoothSilk®/SilkSurface® (Silk), VelvetSurface ® (Velvet), Siltex®, and Biocell®. The roughness and surface area of each material were assessed using non-contact profilometry. Bacterial attachment (2 h) and biofilm formation (24 h) were evaluated for Staphylococcus epidermidis, Pseudomonas aeruginosa, and Ralstonia pickettii over nine independent experiments using a CDC biofilm reactor and viable plate counts (VPCs) as well as confocal scanning laser microscopy. VPCs of the textured implants were compared relative to the Smooth implant. Results Surface areas increased with roughness and were similar among the three least rough implants (Smooth, Silk, and Velvet) and among the roughest implants (Siltex and Biocell). Overall, VPC indicated there was significantly more bacterial attachment and biofilm formation on the Siltex and Biocell implants than the Silk or Velvet implants, although there were differences between species and time points. CSLM confirmed the formation of thicker biofilms on the implants with rougher surface textures. Conclusion This in vitro study confirmed that implant surfaces with rougher texture, resulting in more surface area, harbored greater biofilm loads than those with smoother surfaces.
Bacterial transfer and biofilm formation in needleless connectors in a clinically simulated in vitro catheter model
(Cambridge University Press, 2023-04) Ryder, Marcia; deLancey-Pulcini, Elinor; Parker, Albert E.; James, Garth A.
Objective: Although needleless connectors (NCs) are widely used in clinical practice, they carry significant risk of bloodstream infection (BSI). In this study, we quantified differences in bacterial transfer and biofilm formation between various NCs. Design: Prospective, clinically simulated in vitro experimental study. Methods: We tested 20 NCs in a 5-day clinical simulation of Staphylococcus aureus inoculations onto NC septum surfaces, which were then flushed with saline and cultured for bacterial transfer. Biofilm formation was measured through destructive sampling of the connector-catheter system. Moreover, 8 NC design factors were evaluated for their influence on bacterial transfer and biofilm formation. This study was designed without a disinfection protocol to ascertain the intrinsic risk of each NC. Results: Clave Neutron and MicroClave had the lowest overall mean log density of bacteria in the flush compared to other NCs (P < .05), except there were no statistically significant differences between Clave Neutron, Microclave, SafeTouch, and SafeAccess (P ≥ .05). The amount of biofilm in the NC was positively associated with bacteria in the flush (P < .0005). Among 8 design factors, flow path was most important, with the internal cannula associated with a statistically significant 1 log reduction (LR) in bacteria in the flush (R2 = 49%) and 0.5–2 LR in the connector (R2 = 34%). All factors together best explained bacteria in the flush (R2 = 65%) and biofilm in the connector (R2 = 48%). Conclusions: Bacterial transfer and biofilm formation in the connector-catheter system varied statistically significantly between the 20 NCs, suggesting that NC choice can lower the risk of developing catheter-related BSIs.
Bayesian estimation and uncertainty quantification in models of urea hydrolysis by E. coli biofilms
(Informa UK Limited, 2021-02) Jackson, Benjamin D.; Connolly, James M.; Gerlach, Robin; Klapper, Issac; Parker, Albert E.
Urea-hydrolysing biofilms are crucial to applications in medicine, engineering, and science. Quantitative information about ureolysis rates in biofilms is required to model these applications. We formulate a novel model of urea consumption in a biofilm that allows different kinetics, for example either first order or Michaelis-Menten. The model is fit it to synthetic data to validate and compare two approaches: Bayesian and nonlinear least squares (NLS), commonly used by biofilm practitioners. The shortcomings of NLS motivate the Bayesian approach where a simple Markov Chain Monte Carlo (MCMC) sampler is applied. The model is then fit to real data of influent and effluent urea concentrations from experiments on biofilms of Escherichia coli. Results from synthetic data aid in interpreting results from real data, where first order and Michaelis-Menten kinetic models are compared. The method shows potential for general applications requiring biofilm kinetic information.
Calculating the limit of detection for a dilution series
(Elsevier BV, 2023-05) Sharp, Julia L.; Parker, Albert E.; Hamilton, Martin A.
Aims. Microbial samples are often serially diluted to estimate the number of microbes in a sample, whether as colony-forming units of bacteria or algae, plaque forming units of viruses, or cells under a microscope. There are at least three possible definitions for the limit of detection (LOD) for dilution series counts in microbiology. The statistical definition that we explore is that the LOD is the number of microbes in a sample that can be detected with high probability (commonly 0.95). Methods and results. Our approach extends results from the field of chemistry using the negative binomial distribution that overcomes the simplistic assumption that counts are Poisson. The LOD is a function of statistical power (one minus the rate of false negatives), the amount of overdispersion compared to Poisson counts, the lowest countable dilution, the volume plated, and the number of independent samples. We illustrate our methods using a data set from Pseudomonas aeruginosa biofilms. Conclusions. The techniques presented here can be applied to determine the LOD for any counting process in any field of science whenever only zero counts are observed. Significance and impact of study. We define the LOD when counting microbes from dilution experiments. The practical and accessible calculation of the LOD will allow for a more confident accounting of how many microbes can be detected in a sample.
Community Engaged Cumulative Risk Assessment of Exposure to Inorganic Well Water Contaminants, Crow Reservation, Montana
(2018-01) Eggers, Margaret J.; Doyle, John T.; Lefthand, M. J.; Young, Sara L.; Moore-Nall, Anita L.; Kindness, L.; Medicine, R. O.; Ford, Tim E.; Dietrich, E.; Parker, Albert E.; Hoover, J. H.; Camper, Anne K.
An estimated 11 million people in the US have home wells with unsafe levels of hazardous metals and nitrate. The national scope of the health risk from consuming this water has not been assessed as home wells are largely unregulated and data on well water treatment and consumption are lacking. Here, we assessed health risks from consumption of contaminated well water on the Crow Reservation by conducting a community-engaged, cumulative risk assessment. Well water testing, surveys and interviews were used to collect data on contaminant concentrations, water treatment methods, well water consumption, and well and septic system protection and maintenance practices. Additive Hazard Index calculations show that the water in more than 39% of wells is unsafe due to uranium, manganese, nitrate, zinc and/or arsenic. Most families’ financial resources are limited, and 95% of participants do not employ water treatment technologies. Despite widespread high total dissolved solids, poor taste and odor, 80% of families consume their well water. Lack of environmental health literacy about well water safety, pre-existing health conditions and limited environmental enforcement also contribute to vulnerability. Ensuring access to safe drinking water and providing accompanying education are urgent public health priorities for Crow and other rural US families with low environmental health literacy and limited financial resources.
Comparing the chlorine disinfection of detached biofilm clusters with those of sessile biofilms and planktonic cells in single-and dual-species cultures
(2011-10) Behnke, S.; Parker, Albert E.; Woodall, Dawn; Camper, Anne K.
Although the detachment of cells from biofilms is of fundamental importance to the dissemination of organisms in both public health and clinical settings, the disinfection efficacies of commonly used biocides on detached biofilm particles have not been investigated. Therefore, the question arises whether cells in detached aggregates can be killed with disinfectant concentrations sufficient to inactivate planktonic cells. Burkholderia cepacia and Pseudomonas aeruginosa were grown in standardized laboratory reactors as single species and in coculture. Cluster size distributions in chemostats and biofilm reactor effluent were measured. Chlorine susceptibility was assessed for planktonic cultures, attached biofilm, and particles and cells detached from the biofilm. Disinfection tolerance generally increased with a higher percentage of larger cell clusters in the chemostat and detached biofilm. Samples with a lower percentage of large clusters were more easily disinfected. Thus, disinfection tolerance depended on the cluster size distribution rather than sample type for chemostat and detached biofilm. Intact biofilms were more tolerant to chlorine independent of species. Homogenization of samples led to significantly increased susceptibility in all biofilm samples as well as detached clusters for single-species B. cepacia, B. cepacia in coculture, and P. aeruginosa in coculture. The disinfection efficacy was also dependent on species composition; coculture was advantageous to the survival of both species when grown as a biofilm or as clusters detached from biofilm but, surprisingly, resulted in a lower disinfection tolerance when they were grown as a mixed planktonic culture.
Comparison of quantification methods for an endoscope lumen biofilm model
(Elsevier BV, 2023-12) Haas, Bruno; James, Sarah; Parker, Albert E.; Gagnon, Marie-Claude; Goulet, Noémie; Labrie, Philippe
Biofilm has been implicated in multi-drug resistant organism outbreaks following endoscopic procedures. Automated Endoscope Reprocessors (AER) are devices validated to clean and disinfect endoscopes per applicable standards. The ISO 15883 part 4 standard guides performance testing validation of AERs, including cleaning performance using a biofilm test soil. The standard recommends assessment of biofilm reduction using protein or carbohydrate quantification methods. The aim of this study was to assess the suitability of various quantification methods using the ISO biofilm model. The ISO 15883 part 5 biofilm test soil method was used to grow biofilm within lumens representative of endoscopes channels. The biofilm was then quantified using five methods: Crystal Violet (CV), Colony Forming Units (CFU), Total Organic Carbon (TOC), protein assay with Orthophtalaldehyde (OPA), and protein assay by micro bicinchoninic acid (μBCA). The five methods were statistically analyzed for their ability to assess biofilm reduction on samples accurately and precisely. In addition, the quantification methods were compared to demonstrate statistical equivalency, and thus their suitability for assessing biofilm cleaning performance testing of AERs.
Convergence in variance of Chebyshev accelerated Gibbs samplers
(2014-02) Fox, Colin; Parker, Albert E.
A stochastic version of a stationary linear iterative solver may be designed to converge in distribution to a probability distribution with a specified mean $\mu$ and covariance matrix $A^{-1}$. A common example is Gibbs sampling applied to a multivariate Gaussian distribution which is a stochastic version of the Gauss--Seidel linear solver. The iteration operator that acts on the error in mean and covariance in the stochastic iteration is the same iteration operator that acts on the solution error in the linear solver, and thus both the stationary sampler and the stationary solver have the same error polynomial and geometric convergence rate. The polynomial acceleration techniques that are well known in numerical analysis for accelerating the linear solver may also be used to accelerate the stochastic iteration. We derive first-order and second-order Chebyshev polynomial acceleration for the stochastic iteration to accelerate convergence in the mean and covariance by mimicking the derivation for the linear solver. In particular, we show that the error polynomials are identical and hence so are the convergence rates. Thus, optimality of the Chebyshev accelerated solver implies optimality of the Chebyshev accelerated sampler. We give an algorithm for the stochastic version of the second-order Chebyshev accelerated SSOR (symmetric successive overrelaxation) iteration and provide numerical examples of sampling from multivariate Gaussian distributions to confirm that the desired convergence properties are achieved in finite precision.
Coupon position does not affect Pseudomonas aeruginosa and Staphylococcus aureus biofilm densities in the CDC biofilm reactor
(Elsevier BV, 2024-08) Buckner, Elizabeth; Buckingham-Meyer, Kelli; Miller, Lindsey A.; Parker, Albert E.; Jones, Christopher J.; Goeres, Darla M.
The CDC Biofilm Reactor method is the standard biofilm growth protocol for the validation of US Environmental Protection Agency biofilm label claims. However, no studies have determined the effect of coupon orientation within the reactor on biofilm growth. If positional effects have a statistically significant impact on biofilm density, they should be accounted for in the experimental design. Here, we isolate and quantify biofilms from each possible coupon surface in the reactor to quantitatively determine the positional effects in the CDC Biofilm Reactor. The results showed no statistically significant differences in viable cell density across different orientations and vertical positions in the reactor. Pseudomonas aeruginosa log densities were statistically equivalent among all coupon heights and orientations. While the Staphylococcus aureus cell growth showed no statistically significant differences, the densities were not statistically equivalent among all coupon heights and orientations due to the variability in the data. Structural differences were observed between biofilms on the high-shear baffle side of the reactor compared to the lower shear glass side of the reactor. Further studies are required to determine whether biofilm susceptibility to antimicrobials differs based on structural differences attributed to orientation.
Design and fabrication of biofilm reactors
(2020) Goeres, Darla M.; Pedersen, Stephen; Warwood, B. K.; Walker, Diane K.; Parker, Albert E.; Mettler, Madelyn; Sturman, Paul J.
Laboratory biofilm reactors are tools that researchers use to grow biofilms that exhibit characteristics sufficiently similar to the environment of interest. Numerous biofilm reactors that model various fluid dynamics are described in scientific literature, each with its associated list of advantages and limitations. This chapter focuses on the process used to design and fabricate biofilm reactors with the stated goal of generating a commercial product. The process begins with identifying the environment of interest and key attributes the reactor should include or model. A prototype is then designed, built, and tested in the laboratory. Modifications are made based upon laboratory performance until a design is achieved that is affordable, practical, operationally simple, and relevant and that provides repeatable, convincing results. This process was used to design the industrial surfaces biofilm reactor, developed to model cooling tower biofilms but suitable to study biofilms grown under low shear, high gas transfer, and intermittently wet conditions.
Detection of Microbes in Ice Using Microfabricated Impedance Spectroscopy Sensors
(The Electrochemical Society, 2023-12) Kaiser-Jackson, Lauren B.; Dieser, Markus; McGlennen, Matthew; Parker, Albert E.; Foreman, Christine M.; Warnat, Stephan
During the growth of a polycrystalline ice lattice, microorganisms partition into veins, forming an ice vein network highly concentrated in salts and microbial cells. We used microfabricated electrochemical impedance spectroscopy (EIS) sensors to determine the effect of microorganisms on the electrochemical properties of ice. Solutions analyzed consisted of a 176 μS cm−1 conductivity solution, fluorescent beads, and Escherichia coli HB101-GFP to model biotic organisms. Impedance spectroscopy data were collected at −10 °C, −20 °C, and −25 °C within either ice veins or ice grains (i.e., no veins) spanning the sensors. After freezing, the fluorescent beads and E. coli were partitioned into the ice veins. The corresponding impedance data were discernibly different in the presence of ice veins and microbial impurities. The presence of microbial cells in ice veins was evident by decreased electrical characteristics (electrode polarization between electrode and ice matrix) relative to solid ice grains. Further, this electrochemical behavior was reversed in all bead-doped solutions, indicating that microbial processes influence sensor response. Linear mixed-effects models empirically corroborated the differences in polarization associated with the presence and absence of microbial cells in ice. We show that EIS has the potential to detect microbes in ice and differentiate between veins and solid grains.
Development, standardization, and validation of a biofilm efficacy test: The single tube method
(2019-10) Goeres, Darla M.; Walker, Diane K.; Buckingham-Meyer, Kelli; Lorenz, Lindsey A.; Summers, Jennifer; Fritz, Blaine; Goveia, Danielle; Dickerman, Grace; Schultz, Johanna M.; Parker, Albert E.
Methods validated by a standard setting organization enable public, industry and regulatory stakeholders to make decisions on the acceptability of products, devices and processes. This is because standard methods are demonstrably reproducible when performed in different laboratories by different researchers, responsive to different products, and rugged when small (usually inadvertent) variations from the standard procedure occur. The Single Tube Method (ASTM E2871) is a standard method that measures the efficacy of antimicrobials against biofilm bacteria that has been shown to be reproducible, responsive and rugged. In support of the reproducibility assessment, a six-laboratory study was performed using three antimicrobials: a sodium hypochlorite, a phenolic and a quaternary/alcohol blend, each tested at low and high efficacy levels. The mean log reduction in viable bacteria in this study ranged from 2.32 to 4.58 and the associated reproducibility standard deviations ranged from 0.89 to 1.67. Independent follow-up testing showed that the method was rugged with respect to deviations in sonication duration and sonication power but slightly sensitive to sonicator reservoir degassing and tube location within the sonicator bath. It was also demonstrated that when a coupon was dropped into a test tube, bacteria can splash out of reach of the applied antimicrobials, resulting in substantial bias when estimating log reductions for the products tested. Bias can also result when testing products that hinder the harvesting of microbes from test surfaces. The culmination of this work provided recommended changes to the early version of the standard method E2871-13 (ASTM, 2013b) including use of splashguards and microscopy checks. These changes have been incorporated into a revised ASTM method E2871-19 (ASTM 2019) that is the basis for the first regulatory method (ATMP-MB-20) to substantiate “kills biofilm” claims for antimicrobials registered and sold in the US.
Direct electric current treatment under physiologic saline conditions kills Staphylococcus epidermidis biofilms via electrolytic generation of hypochlorous acid
(2013-02) Sandvik, Elizabeth L.; McLeod, Bruce R.; Parker, Albert E.; Stewart, Philip S.
The purpose of this study was to investigate the mechanism by which a direct electrical current reduced the viability of Staphylococcus epidermidis biofilms in conjunction with ciprofloxacin at physiologic saline conditions meant to approximate those in an infected artificial joint. Biofilms grown in CDC biofilm reactors were exposed to current for 24 hours in 1/10th strength tryptic soy broth containing 9 g/L total NaCl. Dose-dependent log reductions up to 6.7 log10 CFU/cm2 were observed with the application of direct current at all four levels (0.7 to 1.8 mA/cm2) both in the presence and absence of ciprofloxacin. There were no significant differences in log reductions for wells with ciprofloxacin compared to those without at the same current levels. When current exposures were repeated without biofilm or organics in the medium, significant generation of free chlorine was measured. Free chlorine doses equivalent to the 24-hour endpoint concentration for each current level were shown to mimic killing achieved by current application. Current exposure (1.8 mA/cm2) in medium lacking chloride and amended with sulfate, nitrate, or phosphate as alternative electrolytes produced diminished kills of 3, 2, and 0 log reduction, respectively. Direct current also killed Pseudomonas aeruginosa biofilms when NaCl was present. Together these results indicate that electrolysis reactions generating hypochlorous acid from chloride are likely a main contributor to the efficacy of direct current application. A physiologically relevant NaCl concentration is thus a critical parameter in experimental design if direct current is to be investigated for in vivo medical applications.
Drastic hourly changes in hand hygiene workload and performance rates: a multicenter time series analysis
(Elsevier BV, 2024-09) Moore, Lori; Arbogast, James W.; Robbins, Greg; DiGiorgio, Megan; Parker, Albert E.
Background. High hand hygiene (HH) workload is a commonly cited barrier to optimal HH performance. The objective of this study was to assess trends of HH workload as defined by HH opportunities (HHO) and performance rates over different timescales using automated HH monitoring system data. Methods. This multiyear retrospective observational study was conducted in 58 inpatient units located in 10 North American hospitals. HHO and HH rates were analyzed by time series mixed effects general additive model. Results. Median HH rates peaked at 50.0 between 6 and 7 AM with a trough of 38.2 at 5 PM. HHO over hours in a day were the highest at 184 per hospital unit per hour at 10 AM with a trough of 49.0 between 2 and 3 AM. Median rates for day and night shifts were 40.8 and 45.5, respectively (P = .078). Weekend day shift had the lowest median rate (39.4) compared with any other 12-hour shift (P < .1018). The median rates and HHO varied little across days in a week and months. Conclusions. HH workload and performance rates were negatively correlated and changed drastically over hours in a day. Hospitals should consider HH workload in the development and timely delivery of improvement interventions.