Environmental Science and Pollution Research

, Volume 20, Issue 5, pp 2963–2972 | Cite as

Applicability of a modified MCE filter method with Button Inhalable Sampler for monitoring personal bioaerosol inhalation exposure

Research Article


In this study, a “modified” mixed cellulose ester (MCE) filter culturing method (directly placing filter on agar plate for culturing without extraction) was investigated in enumerating airborne culturable bacterial and fungal aerosol concentration and diversity both in different environments. A Button Inhalable Sampler loaded with a MCE filter was operated at a flow rate of 5 L/min to collect indoor and outdoor air samples using different sampling times: 10, 20, and 30 min in three different time periods of the day. As a comparison, a BioStage impactor, regarded as the gold standard, was operated in parallel at a flow rate of 28.3 L/min for all tests. The air samples collected by the Button Inhalable Sampler were directly placed on agar plates for culturing, and those collected by the BioStage impactor were incubated directly at 26 °C. The colony forming units (CFUs) were manually counted and the culturable concentrations were calculated both for bacterial and fungal aerosols. The bacterial CFUs developed were further washed off and subjected to polymerase chain reaction–denaturing gradient gel electrophoresis (DGGE) for diversity analysis. For fungal CFUs, microscopy method was applied to studying the culturable fungal diversity obtained using different methods. Experimental results showed that the performance of two investigated methods varied with sampling environments and microbial types (culturable bacterial and fungal aerosols). For bacterial aerosol sampling, both methods were shown to perform equally well, and in contrast the “modified” MCE filter method was demonstrated to enumerate more culturable fungal aerosols than the BioStage impactor. In general, the microbial species richness (number of gel bands) was observed to increase with increasing collection time. For both methods, the DGGE gel patterns were observed to vary with sampling time and environment despite of similar number of gel bands. In addition, an increase in sampling time from 20 to 30 min was found not to substantially alter the species richness. Regardless of the sampling methods, more species richness was observed in the outdoor environment than the indoor environment. This study described a new personal bioaerosol exposure assessment protocol, and it was demonstrated applicable in monitoring the personal bioaerosol exposure in replace of an Andersen-type impactor.


“Modified” MCE filter culturing method Button-inhalable sampler BioStage impactor Culturable aerosol diversity Bacterial and fungal aerosols Polymerase chain reaction (PCR) Denaturing gradient gel electrophoresis (DGGE) Similarity dendrogram 



This study was supported by the National Science Foundation of China (grants 21277007, 21077005, and 20877004) and National High Technology Research and Development Program of China (grant 2008AA062503).


  1. Abdulamir AS, Yoke TS, Nordin N, Abu Bakar F (2010) Detection and quantification of probiotic bacteria using optimized DNA extraction, traditional and real-time PCR methods in complex microbial communities. Afr J Biotechnol 9:1481–1492Google Scholar
  2. Agranovski IE, Safatov AS, Borodulin AI, Pyankov OV, Petrishchenko VA, Sergeev AN, Agafonov AP, Ignatiev GM, Sergeev AA, Agranovski V (2004) Inactivation of viruses in bubbling processes utilized for personal bioaerosol monitoring. Appl Environ Microbiol 70:6963–6967CrossRefGoogle Scholar
  3. Alvarez AJ, Buttner MP, Stetzenbach LD (1995) PCR for bioaerosol monitoring—sensitivity and environmental interference. Appl Environ Microbiol 61:3639–3644Google Scholar
  4. Andersen AA (1958) New sampler for the collection, sizing, and enumeration of viable airborne parcticles. J Bacteriol 76:471–484Google Scholar
  5. Ayres JG, Forsberg B, Annesi-Maesano I, Dey R, Ebi KL, Helms PJ, Medina-Ramon M, Windt M, Forastiere F (2009) Climate change and respiratory disease: European Respiratory Society position statement. Eur Respir J 34:295–302CrossRefGoogle Scholar
  6. Bernardo ZP, Urban JE, Maghirang RG, Jerez SB, Goodband RD (2002) Assessment of bioaerosols in swine barns by filtration and impaction. Curr Microbiol 44:136–140CrossRefGoogle Scholar
  7. Brachman PS, Erlich R, Eichenwald HF, Cabelli VJ, Kethley TW, Madein SH, Maltman JR, Middlebrook G, Morton JD, Silver IH, Wolfe EK (1964) Standard sampler for assay of airborne microorganisms. Phil Trans Biol Sci 144:1295Google Scholar
  8. Bundy KW, Gent JF, Beckett W, Bracken MB, Belanger K, Triche E, Leaderer BP (2009) Household airborne Penicillium associated with peak expiratory flow variability in asthmatic children. Ann Allergy Asthma Immunol 103:26–30CrossRefGoogle Scholar
  9. Burton NC, Adhikari A, Grinshpun SA, Hornung R, Reponen T (2005) The effect of filter material on bioaerosol collection of Bacillus subtilis spores used as a Bacillus anthracis simulant. J Environ Monit 7:475–480CrossRefGoogle Scholar
  10. Carnelley T, Haldane JS, Anderson AM (1887) The carbonic acid, organic matter, and micro-organisms in air, more especially of dwellings and schools. Phil Trans Biol Sci 178:61–111CrossRefGoogle Scholar
  11. Cox CS, Wathes CM (1995) Bioaerosols handbook. Lewis, Boca RatonGoogle Scholar
  12. Douwes J, Thorne P, Pearce N, Heederik D (2003) Bioaerosol health effects and exposure assessment: progress and prospects. Ann Occup Hyg 3:187–200CrossRefGoogle Scholar
  13. Droogenbroeck VC, Risseghem VM, Braeckman L, Vanrompay D (2009) Evaluation of bioaerosol sampling techniques for the detection of Chlamydophila psittaci in contaminated air. Vet Microbiol 135:31–37CrossRefGoogle Scholar
  14. Durand KTH, Muilenberg ML, Burge HA, Siexas NS (2002) Effect of sampling time on the culturability of airborne fungi and bacteria sampled by filtration. Ann Occup Hyg 46:113–118CrossRefGoogle Scholar
  15. Feller W (1968) An introduction to probability theory and its applications, 3rd edn. Wiley, New YorkGoogle Scholar
  16. Flesch JP, Norris CH, Nugent AE Jr (1967) Calibrating particulate air samplers with monodisperse aerosols: application to the Andersen cascade impactor. Am Ind Hyg Assoc J 28:507–516CrossRefGoogle Scholar
  17. Folmsbee M, Strevett K, Stafford K, Evenson C (2000) The effect of sampling time on the total efficiency of the Andersen microbial sampler: a field study. J Aerosol Sci 31:263–271CrossRefGoogle Scholar
  18. Guan T, Yao M (2010) Use of carbon-nanotube filter in removing bioaerosols. Journal of Aerosol Science 41:611–620CrossRefGoogle Scholar
  19. Guan Y, Zheng BJ, He YQX, Liu L, Zhuang ZX, Cheung CL, Luo SW, Li PH, Zhang LJ, Guan YJ, Butt KM, Wong KL, Chan KW, Lim W, Shortridge KF, Yuen KY, Peiris JSM, Poon LLM (2003) Isolation and characterization of viruses related to the SARS coronavirus from animals in Southern China. Science 302:276–278CrossRefGoogle Scholar
  20. Hameed AAA, Khoder MI, Yuosra S, Osman AM, Ghanem S (2009) Diurnal distribution of airborne bacteria and fungi in the atmosphere of Helwan area, Egypt. Sci Total Environ 407:6217–6222CrossRefGoogle Scholar
  21. Harold CB, Constantine JA, Theodore D (1980) Morphology of plants and fungi. Harper & Row, New York CityGoogle Scholar
  22. Hospodsky D, Yamamoto N, Peccia J (2010) Accuracy, precision, and method detection limits of quantitative PCR for airborne bacteria and fungi. Appl Environ Microbiol 76(21):7004–7012CrossRefGoogle Scholar
  23. Jones W, Morring K, Morey P, Sorenson W (1985) Evaluation of the Andersen viable impactor for single stage sampling. Am Ind Hyg Assoc J 46:294–298CrossRefGoogle Scholar
  24. Li K, Dong S, Wu Y, Yao M (2010) Comparisons of biological contents in the air samples collected from the ground and an altitude of 238 m. Aerobiologia 26:233–244CrossRefGoogle Scholar
  25. Lighthart B, Prier K, Loper GM, Bromenshenk J (2000) Bees scavenge airborne bacteria. Microb Ecol 39:314–321Google Scholar
  26. Mainelis G, Tabayoyong M (2010) The effect of sampling time on the overall performance of portable microbial impactors. Aerosol Sci Tech 44:75–82CrossRefGoogle Scholar
  27. Mastorides SM, Oehler RL, Greene JN, Sinnott JT, Kranik M, Sandin RL (1999) The detection of airborne Mycobacterium tuberculosis using micropore membrane air sampling and polymerase chain reaction. Chest 115:19–25CrossRefGoogle Scholar
  28. Mayl KR (1964) Calibration of a modified Andersen bacterial aerosol sampler. Appl Environ Microbiol 12:37–43Google Scholar
  29. Murray CS, Simpson A, Custovic A (2004) Allergens, viruses, and asthma exacerbations. Proc Am Thorac Soc 1:99–104CrossRefGoogle Scholar
  30. Muyzer G, Dewaal EC, Uitterlinden AG (1993) Profiling of complex microbial-populations by denaturing dradient gel-electrophoresis analysis of polymerase chain reaction-amplified genes-coding for 16s ribosomal-RNA. Appl Environ Microbiol 59:695–700Google Scholar
  31. Perez-Padilla R, de la Rosa-Zamboni D, Ponce de Leon S, Hernandez M, Quiñones-Falconi F, Bautista E, Ramirez-Venegas A, Rojas-Serrano J, Ormsby CE, Corrales A, Higuera A, Mondragon E, Cordova-Villalobos JA, INER Working Group on Influenza (2009) Pneumonia and respiratory failure from swine-origin influenza A (H1N1) in Mexico. N Engl J Med 361:680–689CrossRefGoogle Scholar
  32. Solomon WR (1970) A simplified application of the Andersen sampler to the study of airborne fungus particles. J Allergy Clin Immunol 45:1–13Google Scholar
  33. Stark KDC, Nicolet J, Frey J (1998) Detection of Mycoplasma hypopneumoniae by air sampling with a nested PCR assay. Appl Environ Microbiol 64:543–548Google Scholar
  34. Stewart SL, Grinshpun SA, Willeke K, Terzieva S, Ulevicius V, Donnelly J (1995) Effect of impact stress on microbial recovery on an agar surface. Appl Environ Microbiol 61:1232–1239Google Scholar
  35. Trout D, Bernstein J, Martinez K, Biagini R, Wallingford K (2001) Bioaerosol lung damage in a worker with repeated exposure to fungi in a water-damaged building. Environ Health Perspect 109:641–644CrossRefGoogle Scholar
  36. Wang Z, Reponen T, Grinshpun SA, Gorny RL, Willeke K (2001) Effect of sampling time and air humidity on the bioefficiency of filter samplers for bioaerosol collection. Journal of Aerosol Science 32:661–674CrossRefGoogle Scholar
  37. Weis CP, Intrepido AJ, Miller AK, Cowin PG, Durno MA, Gebhardt JS, Bull R (2002) Secondary aerosolization of viable Bacillus anthracis spores in a contaminated US Senate Office. JAMA 288:2853–2858CrossRefGoogle Scholar
  38. Willeke K, Baron PA (1993) Aerosol measurement: principle, techniquies and applications. Van Nostrand Reinhold, New YorkGoogle Scholar
  39. Wilson IG (1997) Inhibition and facilitation of nucleic acid amplification. Appl Environ Microbiol 63:3741–3751Google Scholar
  40. Wintzingerode VF, Gobel UB, Stackebrandt E (1997) Determination of microbial diversity in environmental samples: pitfalls of PCR-based rRNA analysis. FEMS Microbiol Rev 21:213–229CrossRefGoogle Scholar
  41. Woodward CL, Park SY, Jackson DR, Li XI, Birkhold SG, Pillai SD, Ricke SC (2004) Optimization and comparison of bacterial load and sampling time for bioaerosol detection systems in a poultry layer house. J Appl Poult Res 13:433–442Google Scholar
  42. Wu Y, Yao M (2010) Inactivation of bacteria and fungus aerosols using microwave irradiation. Journal of Aerosol Science 41:682–693CrossRefGoogle Scholar
  43. Xu Z, Yao M (2011) Analysis of culturable bacterial aerosol diversity obtained using different sampling and cultivation methods. Aerosol Sci Tech 45:1143–1153CrossRefGoogle Scholar
  44. Yao M, Mainelis G (2006) Utilization of natural electrical charges on microorganisms for their collection by electrostatic means. Journal of Aerosol Science 37:513–527CrossRefGoogle Scholar
  45. Yao M, Mainelis G (2007) Analysis of portable impactor performance for enumeration of viable bioaerosols. J Occup Environ Hyg 4:514–524CrossRefGoogle Scholar
  46. Yao M, Mainelis G, An HR (2005) Inactivation of microorganisms using electrostatic fields. Environ Sci Technol 39:3338–3344CrossRefGoogle Scholar
  47. Zhen S, Li K, Yin L, Yao M, Zhang H, Chen L, Zhou M, Chen X (2009) A comparison of the efficiencies of a portable BioStage impactor and a Reuter centrifugal sampler (RCS) high flow for measuring airborne bacteria and fungi concentrations. Journal of Aerosol Science 40:503–513CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  1. 1.State Key Joint Laboratory for Environmental Simulation and Pollution Control College of Environmental Sciences and EngineeringPeking UniversityBeijingChina
  2. 2.Department of Reproductive HealthGuangdong Women and Children HospitalGuangzhouChina

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