Quality Sample Collection, Handling, and Preservation for an Effective Microbial Forensics Program

Budowle, Bruce; Schutzer, Steven E.; Burans, James P.; Beecher, Douglas J.; Cebula, Thomas A.; Chakraborty, Ranajit; Cobb, William T.; Fletcher, Jacqueline; Hale, Martha L.; Harris, Robert B.; Heitkamp, Michael A.; Keller, Frederick Paul; Kuske, Cheryl R.; LeClerc, J. Eugene; Marrone, Babetta L.; McKenna, Thomas; Morse, Stephen; Rodrı́guez, Luis L.; Valentine, Nancy B.; Yadev, Jagjit

doi:10.1128/aem.01165-06

Cited by 62 publications

(26 citation statements)

References 29 publications

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“…The aerosolization of P. agglomerans was performed to verify the recovery of Gram-negative bacteria according to the recommendations of Budowle et al (5). The main goal of our study was to establish the range of detection when bioaerosol-contaminated surfaces were swabbed using a commercially available foam spatula.…”

mentioning

confidence: 99%

Use of a Foam Spatula for Sampling Surfaces after Bioaerosol Deposition

Lewandowski

Kozłowska

Szpakowska

et al. 2010

Appl Environ Microbiol

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The present study had three goals: (i) to evaluate the relative quantities of aerosolized Bacillus atrophaeus spores deposited on the vertical, horizontal top, and horizontal bottom surfaces in a chamber; (ii) to assess the relative recoveries of the aerosolized spores from glass and stainless steel surfaces with a polyester swab and a macrofoam sponge wipe; and (iii) to estimate the relative recovery efficiencies of aerosolized B. atrophaeus spores and Pantoea agglomerans using a foam spatula at several different bacterial loads by aerosol distribution on glass surfaces. The majority of spores were collected from the bottom horizontal surface regardless of which swab type and extraction protocol were used. Swabbing with a macrofoam sponge wipe was more efficient in recovering spores from surfaces contaminated with high bioaerosol concentrations than swabbing with a polyester swab. B. atrophaeus spores and P. agglomerans culturable cells were detected on glass surfaces using foam spatulas when the theoretical surface bacterial loads were 2.88 ؋ 10 4 CFU and 8.09 ؋ 10 6 CFU per 100-cm 2 area, respectively. The median recovery efficiency from the surfaces using foam spatulas was equal to 9.9% for B. atrophaeus spores when the recovery was calculated relative to the theoretical surface spore load. Using a foam spatula permits reliable sampling of spores on the bioaerosol-exposed surfaces in a wide measuring range. The culturable P. agglomerans cells were recovered with a median efficiency of 0.001%, but staining the swab extracts with fluorescent dyes allowed us to observe that the viable cell numbers were higher by 1.83 log units than culturable organisms. However, additional work is needed to improve the analysis of the foam extracts in order to decrease the limit of detection of Bacillus spores and Gram-negative bacteria on contaminated surfaces.Surface sampling is performed on a frequent basis in all situations where clean environment monitoring is needed, e.g., in health care facilities and in the pharmaceutical industry and food industry. An anthrax bioterrorist event in the fall of 2001 has emphasized the importance of efficient sampling methods for detection of pathogenic microorganisms on surfaces within intentionally contaminated locations (22). Unfortunately, our knowledge on the most effective sampling methodology as well as the level of confidence we may have in the results obtained by wiping, swabbing, and other sample collection strategies is still limited (1). Moreover, in most of the studies performed so far, bacteria and/or spores were collected from test samples or coupons of various materials, inoculated with a suspension of microorganisms that had been placed and spread over the surface, and then dried (14, 15). This may not mimic the true situation of surface contamination by a pathogen that has been intentionally released. Edmonds et al. (12) recently reported lower swabbing efficiencies of different types of swab materials used for sampling glass, polycarbonate, and vinyl surfaces contaminat...

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mentioning

confidence: 99%

Use of a Foam Spatula for Sampling Surfaces after Bioaerosol Deposition

Lewandowski

Kozłowska

Szpakowska

et al. 2010

Appl Environ Microbiol

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“…Quando si cominciò, poi, a percepire il rischio di un potenziale utilizzo di agenti biologici per un attacco terroristico, i CDCs entrarono a far parte del Laboratory Response Network, una rete organizzata per realizzare una più rapida identificazione degli agenti utilizzati e, quindi, potenziare la risposta a un possibile attacco con queste armi 13,14 . Il Laboratory Response Network doveva raccordare laboratori sanitari locali con laboratori centrali altamente specializzati (i CDCs appunto), classificati a seconda del livello di sicurezza richiesto (Biosafety level, BSL) in ordine crescente come BSL1, BSL2, BSL3 e BSL4, idonei a operare in condizioni di rischio controllate, avvalendosi delle più moderne tecniche di analisi 15 , secondo procedure operative standardizzate e validate in costante aggiornamento [16][17][18] . Ai CDCs fu affidato il compito di realizzare corsi di formazione per gli operatori che dovevano lavorare con i patogeni "a rischio".…”

Section: La Nuova Frontiera Dei Microbial Forensics: Un Network DI Counclassified

Forensic microbiology and bioterrorism risk (Part II)

Nasso

Romolo²

2007

Emerg Care J

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“…In forensic microbiology the same rules apply, but more detailed investigations may be necessary to obtain relevant information about the history of a specimen, environmental conditions, chemical and physical constitution of the matrix, presence of pollen etc. (2). This can be achieved by particle sizing, electron microscopy, analytical chemistry, isotope analysis, and other techniques.…”

Section: Sample Matrix Analysismentioning

confidence: 99%