College of Engineering
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The College of Engineering at Montana State University will serve the State of Montana and the nation by fostering lifelong learning, integrating learning and discovery, developing and sharing technical expertise, and empowering students to be tomorrow's leaders.
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Item 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.Item Reproducibility of antimicrobial test methods(2018-08) Parker, Albert E.; Hamilton, Martin A.; Goeres, Darla M.We review reproducibility results for methods that test antimicrobial efficacy against biofilms, spores and bacteria dried onto a surface. Our review, that included test results for Pseudomonas aeruginosa, Salmonella choleraesuis and Bacillus subtilis, suggests that the level of reproducibility depends on the efficacy of the antimicrobial agent being tested for each microbe and microbial environment. To determine the reproducibility of a method, several laboratories must independently test the same antimicrobial agent using the method. Little variability among the efficacy results suggests good reproducibility. Such reproducibility assessments currently are hampered by the absence of an objective process for deciding whether the variability is sufficiently small. We present a quantitative decision process that objectively determines whether any method that assesses antimicrobial efficacy is reproducible. Because the perception of acceptable reproducibility may differ among stakeholders, the decision process is governed by a stakeholder's specifications that necessarily includes the efficacy of the agents to be tested.Item Evaluation of regulations for monitoring the microbial quality of drinking water(1998) Camper, Anne K.; Hamilton, Martin A.Item Quantifying biofilm structure(1999) Lewandowski, Zbigniew; Webb, D.; Hamilton, Martin A.; Harkin, GaryThis article defines some quantitative parameters for describing the structure of a biofilm. The parameters can be calculated from a two-dimensional cross-sectional image on a plane parallel to the substratum within an in situ biofilm. Such images can be acquired using a confocal scanning laser microscope (CSLM). The parameters will eventually be used for eliciting relationships between the biofilm's structure and its biochemical function, and for computer model evaluation. The results shown here indicate that the structural parameters appear to be reaching steady-state conditions as the biofilm grows to a steady state.Item Color measurement as a means of quantifying surface biofouling(1998-11) Pitts, Betsey; Hamilton, Martin A.; McFeters, Gordon A.; Stewart, Philip S.; Willse, Alan Ray; Zelver, NickLaboratory reactors fitted with removable ceramic porcelain growth surfaces were inoculated with a consortium of biofilm forming environmental isolates. A Minolta colorimeter CR-200 (Minolta Camera Co., Ltd, Ramsey, NJ) was used in conjunction with a specially designed adapter to evaluate the reflective color of the porcelain disks as biofilm accumulated on them. Areal viable cell counts were monitored over a period of eleven days in two separate experiments and direct color measurements of the untreated, microbially fouled test surfaces were collected. This colorimetric assay was both non-destructive and immediate. A strong linear relationship between log cell density and log color change was observed. The Pearson product moment correlation coefficient for all 45 observations combined was r=0.95. Separate regression lines for each experiment were not significantly different (P=0.19). When adjusted for time, the (partial) correlation coefficient between log cell density and log color change was r=0.87, which suggests that the relationship between the two measures can not be explained by their mutual dependence on time. Reflective color measurement provided a rapid, non-destructive and quantitative measure of biofilm accumulation.Item Effects of substratum topography on bacterial adhesion(1998-12) Scheuerman, Teresa R.; Camper, Anne K.; Hamilton, Martin A.The effect of substratum topography on bacterial surface colonization was studied using a chemically homogeneous silicon coupon. “Grooves” 10 μm deep and 10, 20, 30, and 40 μm wide were etched on the coupon perpendicular to the direction of flow. Flow (Re = 5.5) of a bacterial suspension (108cells/ml) was directed through a parallel plate flow chamber inverted on a confocal microscope. Images were collected in real time to obtain rate and endpoint colonization data for each of three strains of bacteria:Pseudomonas aeruginosaand motile and nonmotilePseudomonas fluorescens.A higher velocity experiment (Re = 16.6) and an abiotic control using hydrophilic, negatively charged microspheres were also performed. Using a colloidal deposition expression, the initial rates of attachment were compared.P. aeruginosaattached at a higher rate thanP. fluorescensmot+ which attached at a higher rate thanP. fluorescensmot−. For all bacteria the rate was independent of groove size and was greatest on the downstream edges of the grooves. Only the motile organisms were found in the bottoms of the grooves. A higher fluid velocity resulted in an increase in the initial rate of attachment. In contrast, there was no adhesion of the beads. Attachment of the bacteria appears to be predominated by transport from the bulk phase to the substratum.Item Bacterial characterization of toilet bowl biofilms(1998-08) Pitts, Betsey; Stewart, Philip S.; McFeters, Gordon A.; Hamilton, Martin A.; Willse, Alan Ray; Zelver, NickMethods have been developed and applied for sampling, characterizing and quantifying naturally occurring toilet bowl biofilms. Ceramic porcelain disks mounted in neoprene rubber strips were sealed in place in toilet bowls in three residences in Bozeman, Montana. In each bowl, duplicate strips were placed above, at and below the water level. In 7 consecutive weeks, duplicate disks from each zone in each bowl were removed. Surface biofouling was measured by viable cell areal density. Specific fouling rates were calculated and variability among toilet bowls and water levels was assessed. Specific fouling rates ranged from 0.0 to 0.46d‐1. Average areal cell densities at the end of 7 weeks ranged from 103 to 107cfu cm‐2. The extent of fouling was highest below the water line. Neutralization of the chlorine residual (typically 0.9 mg l‐1) in one toilet did not increase the extent of fouling compared to the controls. Biofilm areal viable cell densities and bowl water viable counts were positively correlated (r = 0.78). The visual threshold for detection of toilet bowl biofilm by the naked eye was approximately 105 cfu cm‐2. In a heavily fouled toilet bowl, the biofilm was up to 20 μm thick. Microorganisms were isolated from the biofilm and identified. Of the 32 organisms that were further characterized, 10 were identified as Pseudomonas, Sphingomonas or Chryseomonas species.Item Measuring antimicrobial effects on biofilm bacteria: From laboratory to field(1999) Zelver, Nick; Hamilton, Martin A.; Pitts, Betsey; Goeres, Darla M.; Walker, Diane K.; Sturman, Paul J.; Heersink, JoannaItem Quantitative analysis of a presence/absence microbiological assay: the hard surface carrier test of disinfectant efficacy(1996-09) Hamilton, Martin A.; DeVries, Todd AlanItem Chemical effects of biofilm colonization on 304 stainless steel(1996-05) Pendyala, Jyostna; Avci, Recep; Geesey, Gill G.; Stoodley, Paul; Hamilton, Martin A.; Harkin, GaryChanges in the surface concentrations of the main alloying elements of an as‐received 304 stainless steel, exposed to a mixed culture of biofilm‐forming bacteria under flowing conditions, were observed using Auger electron spectroscopy. In the oxide film close to the bulk stainless steel, there was an enrichment in the relative concentration of Cr with a corresponding decrease in the relative Fe concentration as compared to a control coupon exposed only to sterile media. There were no changes observed in the relative Ni concentration.