Evaluating Composite Sampling Methods of Bacillus Spores at Low Concentrations

Hess, Becky M.; Amidan, Brett G.; Anderson, Kevin K.; Hutchison, Janine R.; Nychas, George-John

doi:10.1371/journal.pone.0164582

Evaluating Composite Sampling Methods of Bacillus Spores at Low Concentrations

Journal Article · Thu Oct 13 00:00:00 EDT 2016 · PLoS ONE

DOI:https://doi.org/10.1371/journal.pone.0164582· OSTI ID:1332606

Hess, Becky M. ^[1]; Amidan, Brett G. ^[1]; Anderson, Kevin K. ^[1]; Hutchison, Janine R. ^[1]; Nychas, George-John ^[1]

Pacific Northwest National Lab. (PNNL), Richland, WA (United States). National Security Directorate

Restoring facility operations after the 2001 Amerithrax attacks took over three months to complete, highlighting the need to reduce remediation time. The most time intensive tasks were environmental sampling and sample analyses. Composite sampling allows disparate samples to be combined, with only a single analysis needed, making it a promising method to reduce response times. We developed a statistical experimental design to test three different composite sampling methods: 1) single medium single pass composite: a single cellulose sponge samples multiple coupons; 2) single medium multi-pass composite: a single cellulose sponge is used to sample multiple coupons; and 3) multi-medium post-sample composite: a single cellulose sponge samples a single surface, and then multiple sponges are combined during sample extraction. Five spore concentrations of Bacillus atrophaeus Nakamura spores were tested; concentrations ranged from 5 to 100 CFU/coupon (0.00775 to 0.155CFU/cm², respectively). Study variables included four clean surface materials (stainless steel, vinyl tile, ceramic tile, and painted wallboard) and three grime coated/dirty materials (stainless steel, vinyl tile, and ceramic tile). Analysis of variance for the clean study showed two significant factors: composite method (p-value < 0.0001) and coupon material (p-value = 0.0008). Recovery efficiency (RE) was higher overall using the post-sample composite (PSC) method compared to single medium composite from both clean and grime coated materials. RE with the PSC method for concentrations tested (10 to 100 CFU/coupon) was similar for ceramic tile, painted wall board, and stainless steel for clean materials. RE was lowest for vinyl tile with both composite methods. Statistical tests for the dirty study showed RE was significantly higher for vinyl and stainless steel materials, but significantly lower for ceramic tile. These results suggest post-sample compositing can be used to reduce sample analysis time when responding to a Bacillus anthracis contamination event of clean or dirty surfaces.

View Accepted Manuscript (DOE)

Research Organization:: Pacific Northwest National Laboratory (PNNL), Richland, WA (US)

Sponsoring Organization:: US Department of Homeland Security (DHS); USDOE

Grant/Contract Number:: AC05-76RL01830

OSTI ID:: 1332606

Report Number(s):: PNNL-SA--117699; 400904120

Journal Information:: PLoS ONE, Journal Name: PLoS ONE Journal Issue: 10 Vol. 11; ISSN 1932-6203

Publisher:: Public Library of ScienceCopyright Statement

Country of Publication:: United States

Language:: English

References (25)

Composite Sampling Patil, Ganapati P.; Gore, Sharad D.; Taillie, Charles Springer New York, NY https://doi.org/10.1007/978-1-4419-7628-4	book	January 2011
Evaluation of two methods for detection of viable Bacillus anthracis simulant spores in maritime environmental samples Nelson, Scott; Hofacre, Kent; Calfee, M. Worth Environmental Monitoring and Assessment, Vol. 195, Issue 2 https://doi.org/10.1007/s10661-022-10772-8	journal	January 2023
Commitment of bacterial spores to germinate A measure of the trigger reaction Stewart, G. S.; Johnstone, K.; Hagelberg, E. Biochemical Journal, Vol. 198, Issue 1 https://doi.org/10.1042/bj1980101	journal	July 1981
Analysis of broth-cultured Bacillus atrophaeus and Bacillus cereus spores Buhr, T. L.; McPherson, D. C.; Gutting, B. W. Journal of Applied Microbiology, Vol. 105, Issue 5 https://doi.org/10.1111/j.1365-2672.2008.03899.x	journal	November 2008
Decontamination of a hard surface contaminated with Bacillus anthracisΔSterne and B. anthracis Ames spores using electrochemically generated liquid-phase chlorine dioxide (eClO2): eClO2-treated B. anthracis spores Buhr, T. L.; Young, A. A.; Minter, Z. A. Journal of Applied Microbiology, Vol. 111, Issue 5 https://doi.org/10.1111/j.1365-2672.2011.05122.x	journal	September 2011
Laboratory studies on surface sampling of Bacillus anthracis contamination: summary, gaps and recommendations Piepel, G. F.; Amidan, B. G.; Hu, R. Journal of Applied Microbiology, Vol. 113, Issue 6 https://doi.org/10.1111/j.1365-2672.2012.05381.x	journal	September 2012
Test method development to evaluate hot, humid air decontamination of materials contaminated with Bacillus anthracis ∆Sterne and B. thuringiensis Al Hakam spores Buhr, T. L.; Young, A. A.; Minter, Z. A. Journal of Applied Microbiology, Vol. 113, Issue 5 https://doi.org/10.1111/j.1365-2672.2012.05423.x	journal	September 2012
Evaluation of a Wipe Surface Sample Method for Collection of Bacillus Spores from Nonporous Surfaces Brown, G. S.; Betty, R. G.; Brockmann, J. E. Applied and Environmental Microbiology, Vol. 73, Issue 3 https://doi.org/10.1128/aem.01082-06	journal	November 2006
Quality Sample Collection, Handling, and Preservation for an Effective Microbial Forensics Program Budowle, B.; Schutzer, S. E.; Burans, J. P. Applied and Environmental Microbiology, Vol. 72, Issue 10 https://doi.org/10.1128/aem.01165-06	journal	October 2006
False-Negative Rate and Recovery Efficiency Performance of a Validated Sponge Wipe Sampling Method Krauter, Paula A.; Piepel, Greg F.; Boucher, Raymond Applied and Environmental Microbiology, Vol. 78, Issue 3 https://doi.org/10.1128/aem.07403-11	journal	December 2011
Response surface modeling for hot, humid air decontamination of materials contaminated with Bacillus anthracis ∆Sterne and Bacillus thuringiensis Al Hakam spores Prokop, Edward J.; Crigler, John R.; Wells, Claire M. AMB Express, Vol. 4, Issue 1 https://doi.org/10.1186/s13568-014-0021-3	journal	May 2014
Composite Sampling of a Bacillus anthracis Surrogate with Cellulose Sponge Surface Samplers from a Nonporous Surface Tufts, Jenia A. M.; Meyer, Kathryn M.; Calfee, Michael Worth PLoS ONE, Vol. 9, Issue 12 https://doi.org/10.1371/journal.pone.0114082	journal	December 2014
Swab Materials and Bacillus anthracis Spore Recovery from Nonporous Surfaces Rose, Laura; Jensen, Bette; Peterson, Alicia Emerging Infectious Diseases, Vol. 10, Issue 6 https://doi.org/10.3201/eid1006.030716	journal	June 2004
Recovery balance: a method for estimating losses in a Bacillus anthracis spore sampling protocol Da Silva, S. M.; Urbas, A. A.; Filliben, J. J. Journal of Applied Microbiology, Vol. 114, Issue 3 https://doi.org/10.1111/jam.12090	journal	December 2012
Evaluation of a Wipe Surface Sample Method for Collection of Bacillus Spores from Nonporous Surfaces Brown, G. S.; Betty, R. G.; Brockmann, J. E. Applied and Environmental Microbiology, Vol. 73, Issue 3 https://doi.org/10.1128/AEM.01082-06	journal	November 2006
Quality Sample Collection, Handling, and Preservation for an Effective Microbial Forensics Program Budowle, B.; Schutzer, S. E.; Burans, J. P. Applied and Environmental Microbiology, Vol. 72, Issue 10 https://doi.org/10.1128/AEM.01165-06	journal	October 2006
Recovery of Bacillus Spore Contaminants from Rough Surfaces: a Challenge to Space Mission Cleanliness Control Probst, Alexander; Facius, Rainer; Wirth, Reinhard Applied and Environmental Microbiology, Vol. 77, Issue 5 https://doi.org/10.1128/AEM.02037-10	journal	January 2011
Evaluation of a Macrofoam Swab Protocol for the Recovery of Bacillus anthracis Spores from a Steel Surface Hodges, L. R.; Rose, L. J.; Peterson, A. Applied and Environmental Microbiology, Vol. 72, Issue 6 https://doi.org/10.1128/AEM.02923-05	journal	June 2006
National Validation Study of a Cellulose Sponge Wipe-Processing Method for Use after Sampling Bacillus anthracis Spores from Surfaces Rose, Laura J.; Hodges, Lisa; O'Connell, Heather Applied and Environmental Microbiology, Vol. 77, Issue 23 https://doi.org/10.1128/AEM.05377-11	journal	September 2011
False-Negative Rate and Recovery Efficiency Performance of a Validated Sponge Wipe Sampling Method Krauter, Paula A.; Piepel, Greg F.; Boucher, Raymond Applied and Environmental Microbiology, Vol. 78, Issue 3 https://doi.org/10.1128/AEM.07403-11	journal	December 2011
Evaluation of a Macrofoam Swab Protocol for the Recovery of Bacillus anthracis Spores from a Steel Surface Hodges, L. R.; Rose, L. J.; Peterson, A. Applied and Environmental Microbiology, Vol. 72, Issue 6 https://doi.org/10.1128/aem.02923-05	journal	June 2006
National Validation Study of a Cellulose Sponge Wipe-Processing Method for Use after Sampling Bacillus anthracis Spores from Surfaces Rose, Laura J.; Hodges, Lisa; O'Connell, Heather Applied and Environmental Microbiology, Vol. 77, Issue 23 https://doi.org/10.1128/aem.05377-11	journal	September 2011
Composite Sampling Approaches for Bacillus anthracis Surrogate Extracted from Soil France, Brian; Bell, William; Chang, Emily PLOS ONE, Vol. 10, Issue 12 https://doi.org/10.1371/journal.pone.0145799	journal	December 2015
Recovery Efficiency, False Negative Rate, and Limit of Detection Performance of a Validated Macrofoam-Swab Sampling Method with Low Surface Concentrations of Two Bacillus anthracis Surrogates Piepel, Gregory; Hutchison, Janine; Deatherage Kaiser, Brooke https://doi.org/10.2172/1179143	report	March 2015
False Negative Rates of a Macrofoam-Swab Sampling Method with Low Surface Concentrations of Two Bacillus anthracis Surrogates via Real-Time PCR Hutchison, Janine R.; Piepel, Gregory F.; Amidan, Brett G. https://doi.org/10.2172/1186982	report	May 2015

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59 BASIC BIOLOGICAL SCIENCES
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Composite Sampling
spores
statistical test design

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