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dc.contributor.authorParker, Albert E.
dc.contributor.authorMiller, Lindsey
dc.contributor.authorAdams, Jacob
dc.contributor.authorPettigrew, Charles
dc.contributor.authorBuckingham‐Meyer, Kelli
dc.contributor.authorSummers, Jennifer
dc.contributor.authorChristen, Andres
dc.contributor.authorGoeres, Darla
dc.date.accessioned2022-12-07T17:21:36Z
dc.date.available2022-12-07T17:21:36Z
dc.date.issued2022-09
dc.identifier.citationParker, A.E., Miller, L., Adams, J., Pettigrew, C., Buckingham- Meyer, K. & Summers, J. et al. (2022) Imaging and plate counting to quantify the effect of an antimicrobial: A case study of a photo- activated chlorine dioxide treatment. Journal of Applied Microbiology, 133, 3413–3423. Available from: https://doi.org/10.1111/jam.15765en_US
dc.identifier.issn1364-5072
dc.identifier.urihttps://scholarworks.montana.edu/xmlui/handle/1/17459
dc.descriptionThis is the peer reviewed version of the following article: [Imaging and plate counting to quantify the effect of an antimicrobial: A case study of a photo‐activated chlorine dioxide treatment. Journal of Applied Microbiology 133, 6 p3413-3423 (2022)], which has been published in final form at https://doi.org/10.1111/jam.15765. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions: https://authorservices.wiley.com/author-resources/Journal-Authors/licensing/self-archiving.html#3.en_US
dc.description.abstractAim. To assess removal versus kill efficacies of antimicrobial treatments against thick biofilms with statistical confidence. Methods and results. A photo-activated chlorine dioxide treatment (Photo ClO2) was tested in two independent experiments against thick (>100 μm) Pseudomonas aeruginosa biofilms. Kill efficacy was assessed by viable plate counts. Removal efficacy was assessed by 3D confocal scanning laser microscope imaging (CSLM). Biovolumes were calculated using an image analysis approach that models the penetration limitation of the laser into thick biofilms using Beer's Law. Error bars are provided that account for the spatial correlation of the biofilm's surface. The responsiveness of the biovolumes and plate counts to the increasing contact time of Photo ClO2 were quite different, with a massive 7 log reduction in viable cells (95% confidence interval [CI]: 6.2, 7.9) but a more moderate 73% reduction in biovolume (95% CI: [60%, 100%]). Results are leveraged to quantitatively assess candidate CSLM experimental designs of thick biofilms. Conclusions. Photo ClO2 kills biofilm bacteria but only partially removes the biofilm from the surface. To maximize statistical confidence in assessing removal, imaging experiments should use fewer pixels in each z-slice, and more importantly, at least two independent experiments even if there is only a single field of view in each experiment. Significance and impact of study There is limited penetration depth when collecting 3D confocal images of thick biofilms. Removal can be assessed by optimally fitting Beer's Law to all of the intensities in a 3D image and by accounting for the spatial correlation of the biofilm's surface. For thick biofilms, other image analysis approaches are biased or do not provide error bars. We generate unbiased estimates of removal and assess candidate CSLM experimental designs of thick biofilms with different pixilations, numbers of fields of view and number of experiments using the included design tool.en_US
dc.language.isoen_USen_US
dc.publisherWileyen_US
dc.rightscopyright wiley 2022en_US
dc.rights.urihttps://web.archive.org/web/20200106202133/https://onlinelibrary.wiley.com/library-info/products/price-listsen_US
dc.rights.urihttp://web.archive.org/web/20190530141919/https://authorservices.wiley.com/author-resources/Journal-Authors/licensing/self-archiving.htmlen_US
dc.subjectBeer's Lawen_US
dc.subjectbiovolumeen_US
dc.subjectconfocal scanning laser microscopeen_US
dc.subjectexperimental designs for imagingen_US
dc.subjectphoto- activated chlorine dioxideen_US
dc.titleImaging and plate counting to quantify the effect of an antimicrobial: A case study of a photo‐activated chlorine dioxide treatmenten_US
dc.typeArticleen_US
mus.citation.extentfirstpage1en_US
mus.citation.extentlastpage32en_US
mus.citation.issue6en_US
mus.citation.journaltitleJournal of Applied Microbiologyen_US
mus.citation.volume133en_US
mus.identifier.doi10.1111/jam.15765en_US
mus.relation.collegeCollege of Letters & Scienceen_US
mus.relation.departmentMathematical Sciences.en_US
mus.relation.universityMontana State University - Bozemanen_US


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