scholarly journals Evaluation of Two Surface Sampling Methods for Detection of Erwinia herbicola on a Variety of Materials by Culture and Quantitative PCR

2007 ◽  
Vol 73 (11) ◽  
pp. 3505-3510 ◽  
Author(s):  
Mark P. Buttner ◽  
Patricia Cruz ◽  
Linda D. Stetzenbach ◽  
Tracy Cronin
Keyword(s):  
Quantitative Pcr ◽  
Sampling Methods ◽  
Collection Efficiency ◽  
Test Chamber ◽  
Surface Sampling ◽  
Laminate Glass ◽  
Erwinia Herbicola ◽  
Surface Samples ◽  
Sampling Protocols ◽  
Surface Materials

ABSTRACT This research was designed to evaluate surface sampling protocols for use with culture and quantitative PCR (QPCR) amplification assay for detection of the gram-negative bacterial biothreat simulant Erwinia herbicola on a variety of surface materials. Surfaces selected for evaluation were wood laminate, glass and computer monitor screens, metal file cabinets, plastic arena seats, nylon seat cushions, finished concrete flooring, and vinyl tile flooring. Laboratory and test chamber studies were performed to evaluate two sampling methods, a sponge and a macrofoam swab, for detection of E. herbicola on surface materials. In laboratory trials, seven materials were inoculated with a known concentration of E. herbicola cells and samples were collected from the surfaces of the materials to determine sampling efficiencies. Culture analysis was ineffective for assessing E. herbicola collection efficiency because very few culturable cells were obtained from surface samples. QPCR demonstrated that E. herbicola DNA was present in high concentrations on all of the surface samples, and sampling efficiencies ranged from 0.7 to 52.2%, depending on the sampling method and the surface material. The swab was generally more efficient than the sponge for collection of E. herbicola from surfaces. Test chamber trials were also performed in which E. herbicola was aerosolized into the chamber and allowed to settle onto test materials. Surface sampling results supported those obtained in laboratory trials. The results of this study demonstrate the capabilities of QPCR to enhance the detection and enumeration of biocontaminants on surface materials and provide information on the comparability of sampling methods.

scholarly journals Determination of the Efficacy of Two Building Decontamination Strategies by Surface Sampling with Culture and Quantitative PCR Analysis

2004 ◽  
Vol 70 (8) ◽  
pp. 4740-4747 ◽  
Author(s):  
Mark P. Buttner ◽  
Patricia Cruz ◽  
Linda D. Stetzenbach ◽  
Amy K. Klima-Comba ◽  
Vanessa L. Stevens ◽  
...  
Keyword(s):  
Quantitative Pcr ◽  
Chlorine Dioxide ◽  
Pcr Analysis ◽  
Surface Sampling ◽  
Chlorine Dioxide Gas ◽  
Surface Samples ◽  
Qpcr Analysis ◽  
Environmental Background ◽  
Background Material ◽  
Materials Used

ABSTRACT The efficacy of currently available decontamination strategies for the treatment of indoor furnishings contaminated with bioterrorism agents is poorly understood. Efficacy testing of decontamination products in a controlled environment is needed to ensure that effective methods are used to decontaminate domestic and workplace settings. An experimental room supplied with materials used in office furnishings (i.e., wood laminate, painted metal, and vinyl tile) was used with controlled dry aerosol releases of endospores of Bacillus atrophaeus (“Bacillus subtilis subsp. niger,” also referred to as BG), a Bacillus anthracis surrogate. Studies were performed using two test products, a foam decontaminant and chlorine dioxide gas. Surface samples were collected pre- and posttreatment with three sampling methods and analyzed by culture and quantitative PCR (QPCR). Additional aerosol releases with environmental background present on the surface materials were also conducted to determine if there was any interference with decontamination or sample analysis. Culture results indicated that 105 to 106 CFU per sample were present on surfaces before decontamination. After decontamination with the foam, no culturable B. atrophaeus spores were detected. After decontamination with chlorine dioxide gas, no culturable B. atrophaeus was detected in 24 of 27 samples (89%). However, QPCR analysis showed that B. atrophaeus DNA was still present after decontamination with both methods. Environmental background material had no apparent effect on decontamination, but inhibition of the QPCR assay was observed. These results demonstrate the effectiveness of two decontamination methods and illustrate the utility of surface sampling and QPCR analysis for the evaluation of decontamination strategies.

Evaluation of Two Surface Sampling Methods for Microbiological and Chemical Analyses To Assess the Presence of Biofilms in Food Companies

2017 ◽  
Vol 80 (12) ◽  
pp. 2022-2028 ◽  
Author(s):  
Sharon Maes ◽  
Son Nguyen Huu ◽  
Marc Heyndrickx ◽  
Stephanie van Weyenberg ◽  
Hans Steenackers ◽  
...  
Keyword(s):  
Sampling Methods ◽  
Chemical Components ◽  
Chemical Analyses ◽  
Surface Sampling ◽  
Surface Samples ◽  
Attached Bacteria ◽  
Cleaning And Disinfection ◽  
Microbiological Analyses

ABSTRACTBiofilms are an important source of contamination in food companies, yet the composition of biofilms in practice is still mostly unknown. The chemical and microbiological characterization of surface samples taken after cleaning and disinfection is very important to distinguish free-living bacteria from the attached bacteria in biofilms. In this study, sampling methods that are potentially useful for both chemical and microbiological analyses of surface samples were evaluated. In the manufacturing facilities of eight Belgian food companies, surfaces were sampled after cleaning and disinfection using two sampling methods: the scraper–flocked swab method and the sponge stick method. Microbiological and chemical analyses were performed on these samples to evaluate the suitability of the sampling methods for the quantification of extracellular polymeric substance components and microorganisms originating from biofilms in these facilities. The scraper–flocked swab method was most suitable for chemical analyses of the samples because the material in these swabs did not interfere with determination of the chemical components. For microbiological enumerations, the sponge stick method was slightly but not significantly more effective than the scraper–flocked swab method. In all but one of the facilities, at least 20% of the sampled surfaces had more than 102 CFU/100 cm2. Proteins were found in 20% of the chemically analyzed surface samples, and carbohydrates and uronic acids were found in 15 and 8% of the samples, respectively. When chemical and microbiological results were combined, 17% of the sampled surfaces were contaminated with both microorganisms and at least one of the analyzed chemical components; thus, these surfaces were characterized as carrying biofilm. Overall, microbiological contamination in the food industry is highly variable by food sector and even within a facility at various sampling points and sampling times.

scholarly journals Enhanced Detection of Surface-Associated Bacteria in Indoor Environments by Quantitative PCR

2001 ◽  
Vol 67 (6) ◽  
pp. 2564-2570 ◽  
Author(s):  
Mark P. Buttner ◽  
Patricia Cruz-Perez ◽  
Linda D. Stetzenbach
Keyword(s):  
Quantitative Pcr ◽  
Sampling Methods ◽  
Indoor Environments ◽  
Surface Sampling ◽  
Cotton Swab ◽  
Associated Bacteria ◽  
Biological Contamination ◽  
Qpcr Method ◽  
Biological Contaminants ◽  
Flooring Materials

ABSTRACT Methods for detecting microorganisms on surfaces are needed to locate biocontamination sources and to relate surface and airborne concentrations. Research was conducted in an experimental room to evaluate surface sampling methods and quantitative PCR (QPCR) for enhanced detection of a target biocontaminant present on flooring materials. QPCR and culture analyses were used to quantitateBacillus subtilis (Bacillus globigii) endospores on vinyl tile, commercial carpet, and new and soiled residential carpet with samples obtained by four surface sampling methods: a swab kit, a sponge swipe, a cotton swab, and a bulk method. The initial data showed that greater overall sensitivity was obtained with the QPCR than with culture analysis; however, the QPCR results for bulk samples from residential carpet were negative. The swab kit and the sponge swipe methods were then tested with two levels of background biological contamination consisting of Penicillium chrysogenum spores. The B. subtilis values obtained by the QPCR method were greater than those obtained by culture analysis. The differences between the QPCR and culture data were significant for the samples obtained with the swab kit for all flooring materials except soiled residential carpet and with the sponge swipe for commercial carpet. The QPCR data showed that there were no significant differences between the swab kit and sponge swipe sampling methods for any of the flooring materials. Inhibition of QPCR due solely to biological contamination of flooring materials was not evident. However, some degree of inhibition was observed with the soiled residential carpet, which may have been caused by the presence of abiotic contaminants, alone or in combination with biological contaminants. The results of this research demonstrate the ability of QPCR to enhance detection and enumeration of biocontaminants on surface materials and provide information concerning the comparability of currently available surface sampling methods.

A comparison of surface sampling methods for coarse fluvial sediments

1996 ◽  
Vol 32 (10) ◽  
pp. 3219-3226 ◽  
Author(s):  
Ellen E. Wohl ◽  
Deborah J. Anthony ◽  
Susan W. Madsen ◽  
Douglas M. Thompson
Keyword(s):  
Sampling Methods ◽  
Fluvial Sediments ◽  
Surface Sampling

scholarly journals Comparative evaluation of vacuum-based surface sampling methods for collection of Bacillus spores

2013 ◽  
Vol 95 (3) ◽  
pp. 389-396 ◽  
Author(s):  
M. Worth Calfee ◽  
Laura J. Rose ◽  
Stephen Morse ◽  
Dino Mattorano ◽  
Matt Clayton ◽  
...  
Keyword(s):  
Comparative Evaluation ◽  
Sampling Methods ◽  
Surface Sampling ◽  
Bacillus Spores

scholarly journals Comparing different sampling methods in order to reconstruct plant economies at the Eneolithic lake dwelling site Stare gmajne, Slovenia

2016 ◽  
Vol 43 ◽  
pp. 413-420
Author(s):  
Tjaša Tolar ◽  
Anton Velušček
Keyword(s):  
Sampling Methods ◽  
Cultural Layer ◽  
Surface Sampling ◽  
Plant Macroremains ◽  
Plant Remains ◽  
Ljubljansko Barje

The results of plant macroremains studies of the Eneolithic (c. 3160–3100 cal BC) lakeshore settlement at Stare gmajne on the Ljubljansko barje in Slovenia are presented. Archaeobotanical material was collected in two different ways: (1) systematic surface sampling from the cultural layer, and (2) judgement sampling from an incompletely burnt large loom-weight. The preservation state and the spectra of plant macroremains were different in both types of samples. The first study primarily deals with the waterlogged plant remains of various types and taxa, while the second deals with carbonised and half-carbonised cereal macroremains, mostly chaff. Both studies confirm the cultivation of main crops: emmer, einkorn and barley.

scholarly journals Towards precision ecology: Relationships of multiple sampling methods quantifying abundance for comparisons among studies

2021 ◽  
Author(s):  
Benjamin D. Hoffmann ◽  
Magen Pettit
Keyword(s):  
Sampling Methods ◽  
Full Range ◽  
Transformation Method ◽  
Sampling Techniques ◽  
Pitfall Traps ◽  
Multiple Sampling ◽  
Sampling Protocols ◽  
Zero Values ◽  
Relative Utility ◽  
Ant Ecology

ABSTRACTBecause different sampling techniques will provide different abundance values, it is currently difficult to compare results among many studies to form holistic understandings of how abundance influences ant ecology. Using three sampling methods in the same location we found pitfall traps best confirmed A. gracilipes presence recording the fewest zero values (9.1%), card counts were the least reliable (67.1%), and tuna lures were intermediate (30.1%). The abundance of A. gracilipes from card counts ranged from 0 to 20, in pitfall traps from 0 to 325, and the full range of tuna lure abundance scores (0-7) were sampled. We then determined the relationships between these three standard ant sampling techniques for the abundance of yellow crazy ant Anoplolepis gracilipes. Irrespective of the data transformation method, the strongest relationship was between pitfall traps and tuna lures, and the least strong was between pitfall traps and card counts. We then demonstrate the utility of this knowledge by analysing A. gracilipes abundance reported within published literature to show where the populations in those studies sit on an abundance spectrum. We also comment on insights into the relative utility of the three methods we used to determine A. gracilipes abundance among populations of varying abundance. Pitfall traps was the most reliable method to determine if the species was present at the sample level. Tuna lures were predominantly reliable for quantifying the presence of workers, but were limited by the number of workers that can gather around a spoonful of tuna. Card counts were the quickest method, but were seemingly only useful when A. gracilipes abundance is not low. Finally we discuss how environmental and biological variation needs to be accounted for in future studies to better standardise sampling protocols to help progress ecology as a precision science.

scholarly journals European survey and evaluation of sampling methods recommended by the standard EN ISO 18593 for the detection of Listeria monocytogenes and Pseudomonas fluorescens on industrial surfaces

2020 ◽  
Vol 367 (7) ◽  
Author(s):  
Thomas Brauge ◽  
Lena Barre ◽  
Guylaine Leleu ◽  
Stéphane André ◽  
Catherine Denis ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Pseudomonas Fluorescens ◽  
Sampling Methods ◽  
Bacterial Species ◽  
Surface Sampling ◽  
European Survey ◽  
Spoilage Bacteria ◽  
Significant Difference ◽  
Processing Plants ◽  
Survey And Evaluation

ABSTRACT The ready-to-eat products can be contaminated during processing by pathogen or spoilage bacteria, which persist in the industrial environment. Some bacterial species are able to form biofilms which protect them from environmental conditions. To check the bacterial contamination of the surfaces in the food industries, the professionals must regularly use surface sampling methods to detect the pathogen such as Listeria monocytogenes or the spoilage such as Pseudomonas fluorescens. In 2010, we designed and carried out a European survey to collect surface sampling information to detect or enumerate L. monocytogenes in food processing plants. A total of 137 questionnaires from 14 European Union Member States were returned. The outcome of this survey showed that the professionals preferred friction sampling methods with gauze pad, swab and sponges versus contact sampling methods. After this survey, we compared the effectiveness of these three friction sampling methods and the contact plates, as recommended in the standard EN ISO 18593 that was revised in 2018, on the recovery of L. monocytogenes and of P. fluorescens in mono-specie biofilms. This study showed no significant difference between the effectiveness of the four sampling methods to detach the viable and culturable bacterial population of theses mono-specie biofilms.

scholarly journals Comparison of indoor air sampling and dust collection methods for fungal exposure assessment using quantitative PCR

2017 ◽  
Vol 19 (10) ◽  
pp. 1312-1319 ◽  
Author(s):  
Jennie Cox ◽  
Reshmi Indugula ◽  
Stephen Vesper ◽  
Zheng Zhu ◽  
Roman Jandarov ◽  
...  
Keyword(s):  
Exposure Assessment ◽  
Quantitative Pcr ◽  
Indoor Air ◽  
Sampling Methods ◽  
Air Sampling ◽  
Fungal Contamination ◽  
Dust Collection ◽  
Fungal Exposure ◽  
The Many ◽  
Collection Methods

Evaluating fungal contamination indoors is complicated because of the many different sampling methods utilized.

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