Globally, norovirus is the most common gastroenteritis causing pathogen. Annually, norovirus causes 685 million cases of acute gastroenteritis and 200,000 deaths, worldwide. Recent evidence has suggested that norovirus can also be spread via aerosolization; however, an indoor generation source has yet to be determined. We optimized a sampling method for the collection of aerosolized norovirus using murine norovirus (MNV) as a surrogate. Optimization of the sampling method was performed using two bioaerosol samplers (SKC BioSampler and the NIOSH Bioaerosol Cyclone Sampler 251) and two sampling media (Hanks Balanced Salt Solution [HBSS] and Phosphate Buffered Saline [PBS]). Murine norovirus was aerosolized in a bioaerosol chamber and later collected using each sampler/media combination. Collected MNV was quantified using quantitative polymerase chain reaction (qPCR). Intact capsids of MNV were assessed using propidium monoazide dye in combination with qPCR and confirmed with transmission electron microscopy. Ten trials were conducted, with each trial lasting for 30 min. The SKC BioSampler collected a significantly higher concentration of MNV than the NIOSH-251 sampler did (p-value < 0.0001). However, there were no significant differences in the relative percent of MNV that remained viable between both samplers (p-value = 0.2215). The use of HBSS sampling media yielded a higher concentration of MNV than PBS media (p-value = 0.0125). However, PBS media maintained viability at a significantly higher percentage than HBSS media (p-value < 0.0001). The results support the optimization of a sampling method for the collection of aerosolized MNV and possibly norovirus in different sampling environments.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Bailey, E. S., Choi, J. Y., Zemke, J., Yondon, M., & Gray, G. C. (2018). Molecular surveillance of respiratory viruses with bioaerosol sampling in an airport. Tropical Diseases, Travel Medicine and Vaccines, 4(1), 11.
Blachere, F. M., Lindsley, W. G., Pearce, T. A., Anderson, S. E., Fisher, M., Khakoo, R., et al. (2009). Measurement of airborne influenza virus in a hospital emergency department. Clinical Infectious Diseases, 48(4), 438–440.
Bonifait, L., Charlebois, R., Vimont, A., Turgeon, N., Veillette, M., Longtin, Y., et al. (2015). Detection and quantification of airborne norovirus during outbreaks in healthcare facilities. Clinical Infectious Diseases, 61(3), 299–304.
Brooks, J., Tanner, B., Josephson, K., Gerba, C., Haas, C., & Pepper, I. (2005). A national study on the residential impact of biological aerosols from the land application of biosolids. Journal of Applied Microbiology, 99(2), 310–322.
Cao, G., Noti, J., Blachere, F., Lindsley, W., & Beezhold, D. (2011). Development of an improved methodology to detect infectious airborne influenza virus using the NIOSH bioaerosol sampler. Journal of Environmental Monitoring, 13(12), 3321–3328.
Chang, C.-W., Chou, F.-C., & Hung, P.-Y. (2010). Evaluation of bioaerosol sampling techniques for Legionella pneumophila coupled with culture assay and quantitative PCR. Journal of Aerosol Science, 41(12), 1055–1065.
Chow, C. M., Leung, A., & Hon, K. L. (2010). Acute gastroenteritis: From guidelines to real life. Clinical and Experimental Gastroenterology, 3, 97–112.
Fabian, M., Miller, S., Reponen, T., & Hernandez, M. (2005). Ambient bioaerosol indices for indoor air quality assessments of flood reclamation. Journal of Aerosol Science, 36(5–6), 763–783.
Fabian, P., McDevitt, J., & Houseman, E. (2009a). Airborne influenza virus detection with four aerosol samplers using molecular and infectivity assays: Considerations for a new infectious virus aerosol sampler. Indoor Air, 19, 433–441.
Fabian, P., McDevitt, J. J., Lee, W.-M., Houseman, E. A., & Milton, D. K. (2009b). An optimized method to detect influenza virus and human rhinovirus from exhaled breath and the airborne environment. Journal of Environmental Monitoring, 11(2), 314–317.
Farnsworth, J. E., Goyal, S. M., Kim, S. W., Kuehn, T. H., Raynor, P. C., Ramakrishnan, M., et al. (2006). Development of a method for bacteria and virus recovery from heating, ventilation, and air conditioning (HVAC) filters. Journal of Environmental Monitoring, 8(10), 1006–1013.
Girlando, E. M. (2014). Sampling for airborne influenza virus using RNA preservation buffer: a new approach. Iowa: University of Iowa.
Gregoricus, N., Hall, A. J., Lopman, B., Parashar, U. D., Park, G. W., Vinjé, J., et al. (2011). Updated norovirus outbreak management and disease prevention guidelines. Morbidity and Mortality Weekly Report: Recommendations and Reports, 60(3), 1–15.
Hall, A. J., Wikswo, M. E., Manikonda, K., Roberts, V. A., Yoder, J. S., & Gould, L. H. (2013). Acute gastroenteritis surveillance through the national outbreak reporting system, United States. Emerging Infectious Diseases, 19(8), 1305.
Hermann, J. R., Hoff, S. J., Yoon, K.-J., Burkhardt, A. C., Evans, R. B., & Zimmerman, J. J. (2006). Optimization of a sampling system for recovery and detection of airborne porcine reproductive and respiratory syndrome virus and swine influenza virus. Applied and Environmental Microbiology, 72(7), 4811–4818.
Hogan, C., Jr., Kettleson, E., Lee, M. H., Ramaswami, B., Angenent, L., & Biswas, P. (2005). Sampling methodologies and dosage assessment techniques for submicrometre and ultrafine virus aerosol particles. Journal of Applied Microbiology, 99(6), 1422–1434.
Kienlen, L. L. (2015). Comparison of bioaerosol collection methods in the detection of airborne influenza virus. Iowa: University of Iowa.
Kim, S., & Ko, G. (2012). Using propidium monoazide to distinguish between viable and nonviable bacteria, MS2 and murine norovirus. Letters in Applied Microbiology, 55(3), 182–188.
Knowlton, S. D., Boles, C. L., Perencevish, E. N., Diekema, D. J., & Nonnenmann, M. W. (2018). Bioaerosol concentrations generated from toilet flushing in a hospital-based patient care setting. Antimicrobial Resistance & Infection Control, 7(1), 16.
Lee, M., Seo, D. J., Seo, J., Oh, H., Jeon, S. B., Ha, S.-D., et al. (2015). Detection of viable murine norovirus using the plaque assay and propidium-monoazide-combined real-time reverse transcription-polymerase chain reaction. Journal of Virological Methods, 221, 57–61.
Lin, W.-H., & Li, C.-S. (1998). The effect of sampling time and flow rates on the bioefficiency of three fungal spore sampling methods. Aerosol Science and Technology, 28(6), 511–522.
Lindsley, W. G., Blachere, F. M., Thewlis, R. E., Vishnu, A., Davis, K. A., Cao, G., et al. (2010). Measurements of airborne influenza virus in aerosol particles from human coughs. PLoS ONE, 5(11), e15100.
Lindsley, W. G., Schmechel, D., & Chen, B. T. (2006). A two-stage cyclone using microcentrifuge tubes for personal bioaerosol sampling. Journal of Environmental Monitoring, 8(11), 1136–1142.
Lopman, B. A., Steele, D., Kirkwood, C. D., & Parashar, U. D. (2016). The vast and varied global burden of norovirus: Prospects for prevention and control. PLoS Medicine, 13(4), e1001999.
Macher, J., & Willeke, K. (1992). Performance criteria for bioaerosol samplers. Journal of Aerosol Science, 23, 647–650.
Marks, P., Vipond, I., Carlisle, D., Deakin, D., Fey, R., & Caul, E. (2000). Evidence for airborne transmission of Norwalk-like virus (NLV) in a hotel restaurant. Epidemiology & Infection, 124(3), 481–487.
Nelson, C. A., Wilen, C. B., Dai, Y.-N., Orchard, R. C., Kim, A. S., Stegeman, R. A., et al. (2018). Structural basis for murine norovirus engagement of bile acids and the CD300lf receptor. Proceedings of the National Academy of Sciences, 115(39), E9201–E9210.
Nevalainen, A., Pastuszka, J., Liebhaber, F., & Willeke, K. (1992). Performance of bioaerosol samplers: Collection characteristics and sampler design considerations. Atmospheric Environment: Part A: General Topics, 26(4), 531–540.
Patel, M. M., Widdowson, M.-A., Glass, R. I., Akazawa, K., Vinjé, J., & Parashar, U. D. (2008). Systematic literature review of role of noroviruses in sporadic gastroenteritis. Emerging Infectious Diseases, 14(8), 1224.
R Core Team. (2018). R: A language and environment for statistical computing. Vienna: R Core Team.
Randazzo, W., Khezri, M., Ollivier, J., Le Guyader, F. S., Rodríguez-Díaz, J., Aznar, R., et al. (2018). Optimization of PMAxx pretreatment to distinguish between human norovirus with intact and altered capsids in shellfish and sewage samples. International Journal of Food Microbiology, 266, 1–7.
Randazzo, W., López-Gálvez, F., Allende, A., Aznar, R., & Sánchez, G. (2016). Evaluation of viability PCR performance for assessing norovirus infectivity in fresh-cut vegetables and irrigation water. International Journal of Food Microbiology, 229, 1–6.
Stewart, S. L., Grinshpun, S. A., Willeke, K., Terzieva, S., Ulevicius, V., & Donnelly, J. (1995). Effect of impact stress on microbial recovery on an agar surface. Applied and Environmental Microbiology, 61(4), 1232–1239.
Thedell, T., Boles, C. L., Cwiertny, D. W., Brown, G. D., Qian, J., & Nonnenmann, M. W. (2018). Comparisons of a novel air sampling filter material wash buffers and extraction methods in the detection and quantification of influenza virus. bioRxiv. https://doi.org/10.1101/441154.
Tseng, C.-C., & Li, C.-S. (2005). Collection efficiencies of aerosol samplers for virus-containing aerosols. Journal of Aerosol Science, 36(5–6), 593–607.
Uhrbrand, K., Koponen, I. K., Schultz, A. C., & Madsen, A. M. (2018). Evaluation of air samplers and filter materials for collection and recovery of airborne norovirus. Journal of Applied Microbiology, 124(4), 990–1000.
Uhrbrand, K., Schultz, A. C., Koivisto, A., Nielsen, U., & Madsen, A. (2017). Assessment of airborne bacteria and noroviruses in air emission from a new highly-advanced hospital wastewater treatment plant. Water Research, 112, 110–119.
Uhrbrand, K., Schultz, A. C., & Madsen, A. M. (2011). Exposure to airborne noroviruses and other bioaerosol components at a wastewater treatment plant in Denmark. Food and Environmental Virology, 3(3–4), 130–137.
Verreault, D., Moineau, S., & Duchaine, C. (2008). Methods for sampling of airborne viruses. Microbiology and Molecular Biology Reviews, 72(3), 413–444.
Wang, Z., Reponen, T., Grinshpun, S. A., Górny, R. L., & Willeke, K. (2001). Effect of sampling time and air humidity on the bioefficiency of filter samplers for bioaerosol collection. Journal of Aerosol Science, 32(5), 661–674.
Willeke, K., Lin, X., & Grinshpun, S. A. (1998). Improved aerosol collection by combined impaction and centrifugal motion. Aerosol Science and Technology, 28(5), 439–456.
The authors thank Dr. Skip Virgin’s Laboratory at Washington University School of Medicine for graciously donating the required amount of MNV. We acknowledge the National Institute for Occupational Safety and Health for allowing us to use their NIOSH Bioaerosol Cyclone samplers. We would also like the thank Dr. Tom Peters laboratory at the University of Iowa for technical support involving the creation and testing of the bioaerosol chamber. Lastly, we acknowledge Dr. Matthew Nonnenmann’s Lav Lab, including support staff, for laboratory assistance during this project.
This research was supported by a pilot project research training grant from the Heartland Center for Occupational Health and Safety at the University of Iowa. The Heartland Center is supported by Training Grant No. T42OH008491 from the Centers for Disease Control and Prevention/National Institute for Occupational Safety and Health. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Boles, C., Brown, G., Park, J.H. et al. The Optimization of Methods for the Collection of Aerosolized Murine Norovirus. Food Environ Virol 12, 199–208 (2020). https://doi.org/10.1007/s12560-020-09430-4
- Environmental health
- Exposure science
- Industrial hygiene